Cutaneous malignant melanomas in 57 cats: identification of (amelanotic) signet-ring and balloon cell types and verification of their origin by immunohistochemistry, electron microscopy, and in situ hybridization.

Cutaneous malignant melanomas in cats, both melanotic and amelanotic, were diagnosed in 57 of 1.530 skin tumors during the period 1991-1995. All melanomas occurred in domestic shorthaircats of ages 3-19 years (mean = 11.5 years). Postmortem examination was performed on 16 cats. All had metastases in the regional lymph node and several organ systems. The average time of survival after surgical removal of the tumor was 4.5 months. Histologically, five types of melanomas could be distinguished: epithelioid, spindle, mixed, signet-ring, and balloon cell. Whereas all epithelioid, spindle, and mixed epithelioid/spindle cell types showed pigmentation, signet-ring and balloon cell types were often amelanotic. Immunohistochemical examination of the melanomas revealed a positive staining for S-100, vimentin, and neuron-specific enolase. The melanomas were negative for muscle cell markers, except in some of the signet-ring cell melanomas; 13 of 21 tumors showed a weak positive staining for polyclonal desmin. Electron microscopic examination of signet-ring cell melanomas revealed an abundance of intermediate filaments, whereas in some of these tumors a few cells with melanosomes were found. Nonisotopic in situ hybridization for mRNA encoding for tyrosinase verified the melanocytic origin of the amelanotic signet-ring and balloon cell melanomas.

Phenobarbital pretreatment in vivo and in vitro and the effect of hepatotoxicity of d-galactosamine in rat hepatocytes in culture.

Galactosamine (GalN) is a known hepatotoxic compound, acting by depletion of uracil nucleotides. The relation between an active cytochrome P-450 system (CYP) and the hepatotoxicity of GalN was studied in rat hepatocytes that were pretreated with phenobarbital (PB) in vivo or in vitro. A 24-hr in vitro pretreatment of cultured hepatocytes with PB resulted in a significant decrease in GalN toxicity as measured by lactate dehydrogenase (LDH) leakage. Furthermore, GalN treatment resulted in an increase in the activity of the PB-induced forms of CYP (namely CYP 2B1/2) as measured by 7-pentoxyresorufin O-depentylase (PROD) activity. This increase was not found after GalN treatment of microsomes. GalN had no effect on the concentration of the apoenzymes. GalN administration to hepatocytes of in vivo PB-pretreated rats resulted in a similar effect of GalN on the activity of the CYP enzymes but PB in vivo had no effect on GalN toxicity. These results suggest that GalN treatment may result in a significant increase in the specific activity of CYP 2B1/2 enzymes (PROD), without an obvious increase in the amount of PB-induced apoenzymes. This phenomenon was measurable only in intact cells. No direct relation is assumed between the activity of the CYP apoenzymes and the decrease in GalN toxicity after PB treatment. The toxicity of Galn was inhibited by PB treatment in vitro.

Effects of cytokines, bacterial endotoxin and hydrocortisone on primary hepatocyte cultures of rat and hamster.

In vivo an acute phase reaction is characterized by changes in hepatic protein metabolism and modification of liver drug enzymes (cytochrome P-450). In vitro, mediators of an acute phase reaction have been reported to have a similar effect. In the present study, the effects of bacterial endotoxin (lipopolysaccharide, LPS) and cytokines in hepatocyte cultures of rat and hamster were examined. In both species, a significant increase in secretion of specific acute phase protein was measured after 48 hr of exposure to IL-6, TNF-alpha or LPS. LPS and TNF-alpha increased the secretion of serum amyloid A (SAA) in hamster hepatocytes. LPS and IL-6 (but only in the presence of hydrocortisone) increased the secretion of alpha(2)-macroglobulin (alpha(2)M) in rat hepatocytes. An effect on the total concentration of cytochrome P-450 was not found in either species after exposure to glucocorticoids or to LPS and cytokines, although the latter compounds caused a marked increase in secretion of acute phase proteins.