p62/Sequestosome-1: Mapping Sites of Protein-Handling Stress in Canine Cutaneous Mast Cell Tumors.

Canine cutaneous mast cell tumors (MCT) are common, frequently malignant neoplasms that are currently graded histologically for provision of prognostic information. Continuing evidence of subsets of MCT within certain grades (with differing survival times) indicate the need for biomarkers that will facilitate better patient stratification and also provide further information on the biological processes involved in progression. We decided to investigate the expression of p62/sequestosome-1 (p62/SQSTM1), a stress-inducible "hub protein" found in all cell types that shuttles rapidly between the nucleus and cytoplasm and is known to play important roles in protein handling and tumorigenesis. The identity of canine p62/SQSTM1 was confirmed in silico and by validation of a commercial antibody using both Western blotting and functional (pharmaceutical-based) analyses in cell culture. Using immunohistochemistry, 3 patterns of p62 expression were identified based on the predominant intracellular localization, that is, nuclear, mixed (nuclear and cytoplasmic), and cytoplasmic. There was a highly significant association with the 2-tier (Kiupel) grade (P < .0001), with all p62-nuclear immunoreactivity being associated with low grade and most p62-cytoplasmic immunoreactivity (93%) with high grade. Most but not all mixed nuclear-cytoplasmic labeling occurred in low-grade MCT; in other (human) tumor types, this pattern has been interpreted as borderline malignant. These data indicate that there is a shift in protein-handling stress from the nucleus to the cytoplasm in association with increasing malignancy in MCT. Studies to identify the processes and drug-able targets involved in this progression are ongoing.

The effect of a progressive decrease in the circulating blood volume of the dog on the transthoracic impedance.

The correlations between the haemodynamic and transthoracic electrial impedance changes resulting from a progressive reduction in the circulating blood volume were studied in four intact mongrel dogs artificially ventilated with a mixture of halothane in nitrous oxide-oxygen. The cardiac output of the dogs was measured by both the electrical impedance and the fibre optic dye dilution techniques. It was found that significant correlations existed between the blood lloss and the arterial blood pressure, the maximum first derivative of the transthoracic impedance, the Heather Index, the transthoracic impedance, the maximum rate of change of aortic pressure and the cardiac stroke work. There was also a good correlation between the dye and impedance cardiac output values, the impedance value always being higher than the corresponding dye value. The correlation between the Heather Index and the PEP/LVET ratio and 1/PEP2 varied markedly from dog to dog.