Apoptosis inducida por el TNF- (alpha) en células crónicamente infestadas con el virus de la inmunodeficiencia felina

El mecanismo por el cual el virus de la inmunodeficiencia felina (FIV) causa la muerte celular es denominado apoptosis. En este trabajo nosotros inducimos apoptosis con el uso del tumor de necrosis factor alfa (TNF- α) y analizamos el nivel de glutathione (GSH) en una línea celular de fibroblastos felinos (CRFK) crónicamente infectada con FIV. Las células no infectadas permanecieron sin sufrir variación, mientras que las infectadas mostraron un marcado descenso en el nivel de GSH y cambios morfológicos típicos de la apoptosis. El análisis por electroforesis del DNA mostró fragmentación del mismo en pequeñas porciones, indicando el efecto citotóxico del TNF-α en células portadoras del virus. Éste también provocó, a juzgar por la medición de la enzima transcriptasa reversa (RT), un descenso marcado en la replicación viral. Este trabajo tiende no sólo a clarificar la patogénesis, de la infección por FIV, sino también a brindar información para futuras terapias contra esta enfermedad.Palabras claves: Inmunodeficiencia felina, TNF- á apoptosis.AbstractThe mechanism by wich Feline Immunodeficiency Virus (FIV) infection kills the cells is called apoptosis. In this experiment, we induced apoptosis with tumor necrosis factor alfa (TNF- α) and assayed the glutathione (GSH) level, in a feline fibroblastic cell line (CRFK), chronically infected with FIV. Uninfected CRFK cells remained unchanged, while the infected cells showed morphological changes characteristic of apoptosis and a marked decrease in GSH values. DNA electrophoretic assay showed the fragmentation of nucleosomal DNA in the infected cells, indicating the cytocidal effect of TNF-α. In this study, TNF-α was also found to markedly decrease the FIV replication, as shown by the reverse transcriptase (RT) assay. This paper intends to present new information useful for both understanding the pathogenesis of FIV infection and for developing future cures for this disease.Key words: TNF-α, feline immunodeficiency virus apoptosis.

Apoptosis inducida por el TNF- (alpha) en células crónicamente infestadas con el virus de la inmunodeficiencia felina

El mecanismo por el cual el virus de la inmunodeficiencia felina (FIV) causa la muerte celular es denominado apoptosis. En este trabajo nosotros inducimos apoptosis con el uso del tumor de necrosis factor alfa (TNF- α) y analizamos el nivel de glutathione (GSH) en una línea celular de fibroblastos felinos (CRFK) crónicamente infectada con FIV. Las células no infectadas permanecieron sin sufrir variación, mientras que las infectadas mostraron un marcado descenso en el nivel de GSH y cambios morfológicos típicos de la apoptosis. El análisis por electroforesis del DNA mostró fragmentación del mismo en pequeñas porciones, indicando el efecto citotóxico del TNF-α en células portadoras del virus. Éste también provocó, a juzgar por la medición de la enzima transcriptasa reversa (RT), un descenso marcado en la replicación viral. Este trabajo tiende no sólo a clarificar la patogénesis, de la infección por FIV, sino también a brindar información para futuras terapias contra esta enfermedad.Palabras claves: Inmunodeficiencia felina, TNF- á apoptosis.AbstractThe mechanism by wich Feline Immunodeficiency Virus (FIV) infection kills the cells is called apoptosis. In this experiment, we induced apoptosis with tumor necrosis factor alfa (TNF- α) and assayed the glutathione (GSH) level, in a feline fibroblastic cell line (CRFK), chronically infected with FIV. Uninfected CRFK cells remained unchanged, while the infected cells showed morphological changes characteristic of apoptosis and a marked decrease in GSH values. DNA electrophoretic assay showed the fragmentation of nucleosomal DNA in the infected cells, indicating the cytocidal effect of TNF-α. In this study, TNF-α was also found to markedly decrease the FIV replication, as shown by the reverse transcriptase (RT) assay. This paper intends to present new information useful for both understanding the pathogenesis of FIV infection and for developing future cures for this disease.Key words: TNF-α, feline immunodeficiency virus apoptosis.

Detection of Canine Urothelial Carcinoma Cells in Urine Using 5-Aminolevulinic Acid

This study aimed to establish a method to detect canine urothelial carcinoma cells in urine using 5-aminolevulinic acid (5-ALA) and to evaluate its diagnostic accuracy. Urine samples were collected from 21 dogs diagnosed with urothelial carcinoma and three urothelial carcinoma cell lines were used. Urine samples obtained from seven healthy dogs were used as controls. Cells in the urine sediment, or urothelial carcinoma cell lines, were cultured with 5-ALA and then observed under a fluorescence microscope. Moreover, we examined the relationship between fluorescence intensity and the presence of metastasis as well as tumor invasion into the bladder wall in cases of urothelial carcinoma. Urine-derived cells from urothelial carcinoma and urothelial carcinoma cell lines showed clearer red fluorescence with the addition of 5-ALA compared to that exhibited by the cells from healthy dogs. The sensitivity and specificity of the diagnosis of urothelial carcinoma were 90% and 86%, respectively. Significant associations were found between fluorescence intensity and tumor metastasis and bladder wall invasion. This study showed that 5-ALA can be used to detect urothelial carcinoma cells in dogs with relatively high diagnostic accuracy. Further, the fluorescence intensity of tumor cells caused by 5-ALA correlated with the clinical condition of urothelial carcinoma cases, which suggested that 5-ALA could be used as a prognostic marker for canine urothelial carcinoma

Effect of diazoxide on a cat with insulinoma

Case summary The patient was a castrated male American Shorthair cat, approximately 14 years old, weighing 3.4 kg. The patient had chronic kidney disease (CKD) (International Renal Interest Society stages 3–4) as an underlying disease. The cat was examined at a hospital for intermittent lethargy and seizures. Hypoglycaemia was repeatedly observed, and the insulin level was 1.78 ng/ml (reference interval 0.27–0.69) when the blood glucose was 49 mg/dl. Although the cat was tentatively diagnosed with insulinoma, surgery was not recommended because of the severe CKD. Although frequent feeding and prednisolone treatment were initially attempted, blood glucose decreased to 24–42 mg/dl. Diazoxide was additionally prescribed at a dose of 5.2 mg/kg q12h. The cat’s clinical signs improved, and the blood glucose was in the range of 75–103 mg/dl during the first 2 months. It was maintained at >50 mg/dl until the patient died of renal failure 161 days after the start of diazoxide treatment. With regard to adverse events, vomiting once every 2–3 days without weight loss and non-regenerative anaemia were observed, which might have been at least partially caused by diazoxide treatment. An insulinoma was definitively diagnosed via pathological autopsy. Relevance and novel information This is the first reported case of long-term treatment with diazoxide in a cat with insulinoma. Since it was effective in situations where conventional therapies were unsuccessful, diazoxide could be useful as a new therapeutic option for cats with insulinoma. Since adverse events, such as progression of vomiting frequency and non-regenerative anaemia, were observed, careful monitoring was required during administration