RESUMEN
Serum alpha-fetoprotein (AFP) values were measured in hepatic diseased dogs with or without tumor and non-hepatic tumor bearing dogs by a sandwich ELISA using anti-dog AFP antiserum. Serum AFP values were less than 70 ng/ml in clinically healthy dogs. The values in dogs with hepatocellular carcinoma were higher than 1,400 ng/ml in 7 of 9 dogs, wherever those in two dogs with cholangiocarcinoma were in the normal range. Serum AFP values in hepatic diseased dogs without tumor were also high, however, the values were below 500 ng/ml in 90% of the dogs. In non-hepatic tumor dogs, serum AFP values were less than 500 ng/ml in 76% of the dogs. In the surgically removal cases with hepatocellular carcinoma, serum AFP values rapidly decreased. These results suggested that the sandwich ELISA using anti-dog AFP antiserum was an available method for diagnosis of hepatocellular carcinoma in dogs.
Asunto(s)
Carcinoma Hepatocelular/sangre , Colangiocarcinoma/sangre , Hepatopatías/sangre , Neoplasias Hepáticas/sangre , alfa-Fetoproteínas/metabolismo , Animales , Perros , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Masculino , Valores de ReferenciaRESUMEN
OBJECTIVE: To determine the role of platelet activating factor (PAF) in lipopolysaccharide (LPS)-induced thrombocytopenia and neutropenia in dogs. ANIMALS: 42 dogs. PROCEDURES: Blood samples were obtained from dogs given LPS (40 microg/kg of body weight; n = 16), PAF (1 microg/kg; 6), PAF (5 microg/kg/h for 90 minutes; 4), or physiologic saline (0.9% NaCl) solution (0.1 ml/kg/h for 90 minutes; 3) IV to monitor changes in blood cell counts, using automated counters and blood smears stained with Giemsa. Blood samples were also obtained from dogs given LPS (40 microg/kg) that had (n = 5) or had not (6) been treated beforehand with TCV-309, a potent PAF antagonist. Concentration of PAF in blood was determined by use of 125I-radioimmunoassay in dogs given LPS at 1 mg/kg (n = 3) and 40 microg/kg (9). RESULTS: Thrombocytopenia and neutropenia were found in all dogs except those given saline solution. The LPS-induced thrombocytopenia was significantly suppressed by prior treatment with TCV-309. The PAF concentrations increased markedly 1 hour after injection of 1 mg/kg of LPS and increased slightly but significantly 10 minutes after injection of 40 microg/kg of LPS. CONCLUSION AND CLINICAL RELEVANCE: PAF plays an important role in the development of LPS-induced thrombocytopenia and neutropenia in dogs. Control of PAF production, PAF-induced effects, or both may be important in the treatment of dogs with gram-negative bacterial infections and associated thrombocytopenia and neutropenia.
Asunto(s)
Enfermedades de los Perros/etiología , Endotoxemia/veterinaria , Neutropenia/veterinaria , Factor de Activación Plaquetaria/metabolismo , Tetrahidroisoquinolinas , Trombocitopenia/veterinaria , Animales , Pruebas de Coagulación Sanguínea/veterinaria , Perros , Endotoxemia/complicaciones , Femenino , Isoquinolinas/farmacología , Recuento de Leucocitos/veterinaria , Lipopolisacáridos/farmacología , Masculino , Neutropenia/etiología , Proyectos Piloto , Factor de Activación Plaquetaria/farmacología , Inhibidores de Agregación Plaquetaria/farmacología , Recuento de Plaquetas/veterinaria , Compuestos de Piridinio/farmacología , Radioinmunoensayo/veterinaria , Trombocitopenia/etiologíaRESUMEN
Canine alpha-fetoprotein (AFP) was purified by a two step method. Anti-dog AFP antiserum was produced by immunizing rabbits with canine fetal serum proteins that failed to bind to an anti-dog whole adult serum affinity column. Canine AFP was then purified from amniotic fluid using affinity chromatography with anti-dog AFP antiserum. The bound protein was then eluted and further purified by passage through an anti-dog whole adult serum column. The non-binding protein's purity and specificity was confirmed by immunoelectrophoresis, double-diffusion, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and cross-reactivity with anti-human AFP. The molecular weight of canine AFP was approximately 66,000 by SDS-PAGE. Normal adult dogs had serum AFP levels of 7-63 ng ml-1. Levels of AFP were not altered by pregnancy but did show a small peak 2 days following parturition. Newborn puppies had serum AFP levels of 14.08 +/- 5.94 mg ml-1 at birth. By 1 week of age, serum AFP had fallen to 0.766 +/- 0.758 mg ml-1. AFP values in newborn puppies are thus considerably higher than those previously reported in humans, pigs and cattle.
Asunto(s)
Perros/sangre , alfa-Fetoproteínas/análisis , alfa-Fetoproteínas/aislamiento & purificación , Líquido Amniótico/química , Animales , Animales Recién Nacidos/sangre , Cromatografía de Afinidad/veterinaria , Reacciones Cruzadas , Perros/embriología , Electroforesis en Gel de Poliacrilamida/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Sangre Fetal/química , Inmunoelectroforesis/veterinaria , Masculino , Peso Molecular , Embarazo , Conejos , Valores de ReferenciaRESUMEN
An assay procedure for ornithine carbamoyl transferase (OCT), which is known to be a liver-specific enzyme, was modified to be adaptable to bovine serum. The concentration of carbamoyl phosphate and the pH of the reaction reagent solution shifted to 60 mM and 7.2, respectively. These modifications contributed to the augmentation of assay sensitivity. The day-to-day reproducibility was 7.9% (coefficient variation) for serum with high OCT activity and 14.8% for normal bovine serum. The activity was stable for 3-4 months by the storage at -20 degrees C and at least 6 months at -80 degrees C. The OCT activity and other biochemical components including aspartate aminotransferase and gamma glutamyl transpeptidase were measured in sera from 164 dairy or raising cows reared at 7 farms. The normal level of OCT activity in sera from these cows ranged from 9.2 to 25.1 U/l (mean +/- 2SD). By comparison between the farms, the highest mean value of OCT activity was found in a farm where cows were suspected to have some kind of latent liver disease from the data of other biochemical parameters. We conclude that the OCT activity is a very useful diagnostic indicator of liver disease in cattle.