A novel point mutation in the ß1-tubulin gene in asymptomatic macrothrombocytopenic Norfolk and Cairn Terriers.

BACKGROUND: An asymptomatic macrothrombocytopenia, phenotypically similar to asymptomatic inherited macrothrombocytopenia in Cavalier King Charles Spaniels, was described in a group of Norfolk Terriers (NT) from Northern Italy, and isolated cases were also reported in Cairn Terriers (CT). OBJECTIVES: The purpose of this work was to evaluate for the presence of a genetic defect in the ß1-tubulin gene in macrothrombocytopenic NT and CT. METHODS: Samples from 20 healthy dogs (13 NT and 7 CT) were collected at different institutions in Italy (n = 8), United Kingdom (n = 3), and United States (n = 9). Genomic DNA was harvested from EDTA-anticoagulated blood and all coding areas and exon-intron splice sites in the gene encoding ß1-tubulin were amplified and sequenced. RESULTS: Twelve dogs (9 NT and 3 CT) showed a single nucleotide polymorphism (SNP) in exon 1 at nucleotide position 5 (G5A) that would result in the change of an arginine to a histidine at amino acid position 2 (R2H). Four dogs (3 NT and one Cairn Terrier) were heterozygous for the SNP, and 4 dogs (one Norfolk Terrier and 3 CT) matched the normal canine genome. Homozygous dogs for the SNP were macrothrombocytopenic with platelet counts ranging from 19,000 to 110,000/µL. Heterozygous and normal dogs had normal platelet counts and morphology. None had the CKCS point mutation. CONCLUSIONS: The ß1-tubulin N-terminal amino acids form the nucleotide-binding domain and thus this mutation could affect GTP binding enough to influence platelet formation in homozygous but not in heterozygous dogs. The presence of macrothrombocytopenia only in homozygous affected dogs reveals an association between the SNP and the phenotype.

Common bile duct obstruction due to a duodenal gastrinoma in a dog.

In dogs gastrinomas are rare endocrine neoplasms that have always been reported to arise from the pancreas. We report here what we believe to be the first case of a duodenal gastrinoma in a dog. A nine-year-old, male, Pekinese dog was presented with a three-day history of anorexia, vomiting and mucous diarrhoea. Clinical examination and laboratory findings suggested the presence of a severe hepatobiliary disorder. Abdominal ultrasonography showed a diffuse increase in echogenicity of the liver, with severe gallbladder dilation and marked dilation of the cystic duct, common bile duct and extrahepatic bile ducts. Based on these findings, an extrahepatic biliary tract obstruction (EBTO) of unknown cause was suspected. At laparotomy, the gallbladder and the extrahepatic bile ducts appeared severely dilated. The gallbladder was tense and could not be compressed suggesting an outflow obstruction. The duodenum at the level of the common duct orifice appeared slightly thickened and severely hardened for a length of 1 cm. Biopsies from the duodenum and liver were obtained and a cholecystoduodenostomy was performed. The duodenal biopsy revealed severe fibrosis of the submucosa and a infiltrate of small pockets and cords of round to polygonal cells with granular cytoplasm. Based on this appearance the differential diagnoses included neuroendocrine tumours and poorly differentiated carcinoma. Despite surgery and supportive therapy the dog continued to be anorexic and to vomit 3-6 times daily. After euthanasia and necropsy, histopathology showed the presence of a neuroendocrine neoplasia involving the duodenal wall with focal invasion of the adjacent pancreas and small liver metastases. On immunohistochemistry, the cytoplasm of approximately 90% of neoplastic cells intensely expressed neuron specific enolase and gastrin. These findings were consistent with a diagnosis of gastrinoma.