Immunohistochemical and clinical evaluation of p53 in canine cutaneous mast cell tumors.

One hundred twenty-six cutaneous mast cell tumors obtained by excisional biopsy from 106 dogs were evaluated using immunohistochemical staining for the presence of p53 protein. A standard avidin-biotin immunohistochemical protocol was used incorporating a polyclonal antibody of rabbit origin (CM-1) as the primary antibody. Histopathologic grading of tumors was performed on hemotoxylin and eosin-stained samples. There was a significant difference in the percentage of cells staining positive for p53 for the histopathologic grades (P = 0.0005). Grade III tumors had a significantly greater p53 content than did grade I or II tumors (P < 0.05). Clinical data obtained retrospectively was available for 54 dogs. Tumor recurred in 19 of 54 (35.2%) dogs. Twenty-nine dogs died by the end of the study; 9 of 29 (31.0%) died of mast cell tumor disease. Histopathologic grade showed a significant negative association with survival time. Both clinical stage and histopathologic grade showed a significant negative association with time to recurrence. The percentage of cells staining positive for p53 did not significantly improve the forward analysis. Immunohistochemical detection of p53 did not appear useful in characterizing the clinical association between cutaneous mast cell tumor cellular features and survival time or time to tumor recurrence in dogs.

Isolation and characterization of the canine melanoma antigen recognized by the murine monoclonal antibody IBF9 and its distribution in cultured canine melanoma cell lines.

OBJECTIVE: To characterize the canine melanoma antigen recognized by the murine monoclonal antibody IBF9 as to its cellular location, molecular size, protein and glycogen contents, and distribution in cell lines. SAMPLE POPULATION: 7 cultured canine melanoma cell lines. PROCEDURE: Molecular characteristics of the antigen were determined by western blotting, enzymatic digestion studies, and tunicamycin inhibition studies. Distribution of the antigen in the cultured melanoma cell lines was determined by flow cytometry. RESULTS: The antigen consists of 2 proteins with molecular mass of 89 and 85 kd. Tunicamycin and enzymatic digestion studies indicated that these proteins contained little glycosylation. Immunogold and immunofluorescence studies localized the antigen to the cell surface. Antigen expression was consistent within each cell line, with > 90% of the cells positive for all cell lines except 1 (80%). Percentage of positive cells and relative intensity of immunostaining were constant throughout all phases of the cell cycle. CONCLUSIONS: The antigen identified by MAB IBF9 is a well-conserved and highly expressed cell surface protein present during all phases of the cell cycle in all malignant canine melanoma cell lines examined. CLINICAL RELEVANCE: Because of consistency in expression, the antigen may have potential for use in dogs for melanoma immunodiagnostics and immunotherapy.

Megakaryoblastic leukemia in a dog.

A 7-year-old spayed Louisiana Catahoula Leopard dog was examined to determine the cause of shifting forelimb lameness, anorexia, and lethargy. The dog was pyrectic and had splenomegaly, thrombocytopenia, and nonregenerative anemia. Examination of a bone marrow aspirate revealed hypocellularity with normal maturation of erythroid and granulocytic cell lines; however, approximately half of the cells were large undifferentiated blast cells. These cells were identified as megakaryoblasts, using immunohistochemical techniques to detect reactivity for Factor VIII-related antigen and platelet glycoprotein IIIa. Necropsy revealed diffuse neoplastic involvement of the spleen, liver, lungs, bone marrow, and lymph nodes. Cellular infiltrate was characterized by a mixture of megakaryoblasts and typical megakaryocytes. Megakaryoblastic leukemia (M7) is the designation proposed by the Animal Leukemia Study Group for myeloproliferative neoplasms of megakaryocytic lineage.

Effect of deferoxamine and hyperbaric oxygen on free, autogenous, full-thickness skin grafts in dogs.

Free, autogenous, full-thickness skin grafts were applied to 10 dogs; 5 dogs were given an iron chelator, deferoxamine-10% hydroxyethyl pentafraction starch (DEF-HES; 50 mg/kg of body weight, IV), and 5 dogs were given an equal volume of 10% hydroxyethyl pentafraction starch (HES) in 0.9% saline solution (5 ml/kg, IV). All dogs (DEF-HES/HBO- and HES/HBO-treated) were exposed to 60 minutes of hyperbaric oxygen (HBO) at 2 atmospheres absolute pressure twice daily for 10 days, beginning the day of surgery. The percentage of viable graft on day 10 was lower in HES/HBO-treated dogs (mean +/- SD, 13.3 +/- 21.3%; median, 3.0%) than in DEF-HES/HBO-treated dogs (64.7 +/- 39.2%; 88.3%; P = 0.095, Mann-Whitney two-tailed test). There was a positive correlation between percentage of viable graft (on day 10) and percentage of haired skin on the graft site (on day 28) for all dogs (r = 0.91) and for HES/HBO-treated dogs (r = 0.97). The DEF-HES/HBO-treated dogs had less consistent correlation (r = 0.67). Perivascular aggregates of foamy cells were observed in the superficial and reticular portions of the dermis and in the subcutaneous tissue on both surfaces of the panniculus muscle in the graft sites of DEF-HES/HBO-treated dogs. These cells were also observed in the dermis, but not subcutaneous tissue of the control skin sections, and in some viscera of DEF-HES/HBO-treated dogs. Deferoxamine appears to attenuate the detrimental effect of HBO and HES on survival of free skin grafts.(ABSTRACT TRUNCATED AT 250 WORDS)

Malignant retroperitoneal paraganglioma in a horse.

A large primary retroperitoneal sublumbar neoplasm in a horse, with disseminated neoplastic foci in the brain, lung, kidney and spleen is described. The diagnosis was based on light microscopical studies and positive immunostaining for neuron-specific enolase. Because of the location of the primary tumour mass, the aortico-sympathetic ganglion (organ of Zukerkandl) is proposed as the origin.

Effect of deferoxamine-hydroxyethyl pentafraction starch on free, autogenous full-thickness skin grafts in dogs.

Free, autogenous, full-thickness skin grafting was performed on 10 dogs; 5 dogs were given an iron chelator, deferoxamine-10% hydroxyethyl pentafraction starch (DEF-HES; 50 mg/kg of body weight, IV), and 5 dogs were given 10% hydroxyethyl pentafraction starch (HES) in 0.9% saline solution (5 ml/kg, IV). The percentage of viable graft on day 10 was higher, but not significantly so, in DEF-HES-treated dogs (mean +/- SD, 72.6 +/- 24.8%; median 76.5%) than in HES-treated dogs (mean +/- SD, 46.7 +/- 34.3%; median, 48.8%). A trend for a positive correlation between the percentage of viable graft (on day 10) and the percentage of original graft area (on day 28) was observed in HES- and DEF-HES-treated dogs; this trend was significant in HES-treated dogs (P = 0.012). Both groups had significant positive correlation between percentage of viable graft on day 10 and percentage of haired skin on day 28 (HES, P = 0.000002; DEF-HES, P = 0.0148). A unique finding in DEF-HES treated dogs was the consistent observation of foamy macrophages in the dermis adjacent to the grafts, in deep subcutaneous tissue below the grafts, and in normal dermis.

A multichromatic stain for Lowicryl K4M embedded tissues.

A staining procedure using celestine blue and chromotrope 2R for Lowicryl K4M embedded tissues is presented. The stain produces a reliable multichromatic stain for light microscopy on Lowicryl embedded semithin sections.