Kazushi Asano

Validation of the prognostic value of histopathological grading or c-kit mutation in canine cutaneous mast cell tumours: a retrospective cohort study.

The objective of this retrospective cohort study was to validate the prognostic value of histological grading of canine cutaneous mast cell tumours (MCTs) according to the Patnaik (grades I-III) and Kiupel (low, high) grading systems, and to confirm the prognostic significance of internal tandem duplications (ITDs) within exon 11 of the c-kit gene (ITD-Exon11). The baseline characteristics and outcome data from 47 dogs diagnosed with cutaneous MCTs were collected and reviewed. Tumours were graded according to both grading systems and the nucleotide sequence of c-kit was evaluated. Results were analyzed to evaluate predictive factors for overall survival (OS) and progression-free survival (PFS). Log-rank tests indicated that dogs with Patnaik grade III MCTs had significantly reduced OS and PFS compared to those with either grade I or II tumours. However, no significant difference in OS or PFS was observed between grade I and II tumours. The dogs with Kiupel high-grade MCTs had significantly shorter OS and PFS than dogs with low-grade MCTs. The presence of ITD-Exon11 was significantly associated with shorter PFS. The result of Cox regression analysis showed that the Kiupel grading system for OS and PFS, and lymph node metastasis for OS, independently predicted prognosis. Kappa statistics confirmed a significantly higher inter-observer consistency for the Kiupel compared to the Patnaik grading system. These findings demonstrate that the Kiupel grading system is a useful prognostic tool for canine cutaneous MCTs in predicting OS and PFS, while the occurrence of ITD-Exon11 appeared to be a useful predictor for PFS.

Triple-phase helical computed tomography in dogs with hepatic masses.

The purpose of this study was to determine the utility of triple-phase helical computed tomography (CT) for differentiating canine hepatic masses. Seventy dogs with hepatic masses underwent triple-phase CT followed by surgical removal of the hepatic masses. Triple-phase helical CT scans for each dog included precontrast, arterial phase, portal venous phase, and delayed phase studies. The removed hepatic masses were histopathologically classified as hepatocellular carcinoma (n = 47), nodular hyperplasia (n = 14), and hepatic metastatic tumors (n = 9) in dogs. Of the 47 hepatocellular carcinomas, the most common CT findings included a heterogeneous pattern with hyper-, iso-, and hypoenhancement in both the arterial and portal venous phases (40/47, 85.1%). Of the 14 nodular hyperplasias, the most common CT findings were a homogeneous pattern with hyper- and isoenhancement in both the portal venous and delayed phases (13/14, 92.9%). Of nine hepatic metastatic tumors, the most common CT findings included a homogeneous hypoenhancement pattern in both the arterial and portal venous phases (8/9, 88.9%). In addition, 5 (55.6%) showed homogeneous hypoenhancement patterns in the delayed phase. Findings from our study indicated that triple-phase CT is a useful tool for preoperative differentiation of hepatocellular carcinoma, nodular hyperplasia, and hepatic metastatic tumors in dogs.

Plasma thrombin- antithrombin complex concentrations in dogs with malignant tumours

IT has been suggested that coagulation abnormalities such as disseminated intravascular coagulation (DIC) might be caused by the excessive activation of coagulation in dogs with malignant tumours (O’Keefe and Couto 1988). Since excessive activation of coagulation leads to over-generation of thrombin, increased concentrations of thrombin measured in circulating blood would indicate an activation of coagulation. Thrombin-antithrombin complexes (TATs), which are formed rapidly after thrombin production, have been identified as a marker of coagulation activation (Pelzer and others 1988). The plasma TAT concentration has been reported to be useful for evaluating the activation of coagulation in dogs (Ravanat and others 1995), and is available as a marker of the hypercoagulable state in dogs with Cushing’s syndrome (Jacoby and others 2001). However, the plasma TAT concentrations in dogs with malignant tumours have not been reported. This short communication describes the plasma TAT concentrations in dogs with benign or malignant tumours and the incidence of a hypercoagulable state in dogs with malignant tumours. The plasma TAT concentrations of three groups of dogs were examined. Group 1 comprised 16 clinically healthy adult dogs; the dogs were considered clinically normal on the basis of physical examination, routine haematological examination and serum biochemical analysis. Group 2 comprised 11 dogs with benign tumours: five with an adenoma, three with a leiomyoma, two with a haemangioma and one with a lipoma. Group 3 consisted of 62 dogs with malignant tumours, this group was further divided into four subgroups: 27 dogs with epithelial tumours, 17 with mesenchymal tumours except haemangiosarcoma; 10 with haemangiosarcomas and eight with haematopoietic tumours. The dogs in groups 2 and 3 were referred to the Animal Medical Center of Nihon University and their tumours were diagnosed by histopathological examination. None of the dogs received any anticoagulants or blood products before blood sampling. Blood samples were collected into tubes containing 0·13M trisodium citrate (nine parts blood to one part anticoagulant) and centrifuged at 2000 g for 10 minutes, and the citrated plasma was frozen at –30°C until analysis. The plasma TAT concentrations were measured by enzyme immunoassay (TAT Test Kokusai-F; International Reagents Corporation). Mann-Whitney U tests were used to compare plasma TAT concentrations between the groups. For statistical analysis, plasma TAT concentrations undetectable by the assay method used (<0·4 ng/ml) were regarded as 0·4 ng/ml. To detect the incidence of a hypercoagulable state, a reference range of plasma TAT concentration was established as the mean (2sd) concentration obtained from group 1. Dogs with plasma TAT concentrations above the reference range were regarded as displaying a hypercoagulable state. The median (range) plasma TAT concentrations were 0·5 (<0·4 to 0·6) ng/ml in group 1, 0·4 (<0·4 to 6·3) ng/ml in group 2 and 1·3 (0·4 to 49·3) ng/ml in group 3; concentrations were significantly higher in group 3 than in group 1 (P<0·0001). However, there was no significant difference in the plasma TAT concentrations of groups 1 and 2 (P=0·8823) (Fig 1). Within group 3, the TAT concentrations were 0·8 (0·4 to 3·8) ng/ml for the dogs with epithelial tumours, 0·7 (0·4 to 49·3) ng/ml for those with mesenchymal tumours, 17·3 (4·4 to 42·5) ng/ml for those with haemangiosarcomas, and 8·2 (2·1 to 19·0) ng/ml for those with haematopoietic tumours. The plasma TAT concentrations were significantly elevated in all subgroups of group 3 (P<0·001, P<0·01, P<0·0001 and P<0·0001, respectively) compared with group 1 (Fig 2). The mean (2sd) plasma TAT concentration of dogs in group 1 was 0·49 (0·12) ng/ml, and thus dogs with plasma TAT concentrations in excess of 0·61 ng/ml were considered as disVeterinary Record (2005) 156, 839-840

Evaluation of bone deformities of the femur, tibia, and patella in Toy Poodles with medial patellar luxation using computed tomography

OBJECTIVES To evaluate morphological parameters of the femur, tibia, and patella in Toy Poodles with medial patellar luxation (MPL) using three-dimensional (3D) computed tomography (CT) and to compare these parameters between radiography and CT. METHODS Thirty-five hindlimbs of Toy Poodles were divided into normal and grade 2 and 4 MPL groups. The anatomical and mechanical lateral proximal femoral angle, anatomical and mechanical lateral distal femoral angle (aLDFA, mLDFA), femoral varus angle (FVA), inclination of the femoral head angle, procurvation angle, anteversion angle (AA), frontal angle of the femoral neck, mechanical medial proximal or distal tibial angle, mechanical cranial proximal or distal tibial angle, tibial plateau angle, tibial torsion angle (TTA), Z angle, relative tibial tuberosity width, ratio of the medial distance of tibial tuberosity to the proximal tibial width (MDTT/PTW), patella size, and the patellar ligament length: patellar length (L:P) ratio were evaluated on radiography and 3D CT. RESULTS The aLDFA, mLDFA, FVA, and TTA were significantly larger and the AA, MDTT/PTW, and patella were significantly smaller in the grade 4 MPL group. There were significant differences in many parameters between imaging tools, and CT was considered less susceptible to potential artefacts and rotational deformities. CLINICAL SIGNIFICANCE Toy Poodles with grade 4 MPL had significant femoral varus deformity, medial displacement of the tibial tuberosity, internal torsion of the proximal tibia, and hypoplasia of the patella.

Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca2+ mobilization of the cells was evaluated using the Ca2+ indicator Fluo3 to monitor Ca2+ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca2+ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.

Intraoperative identification of canine hepatocellular carcinoma with indocyanine green fluorescent imaging.

OBJECTIVES To evaluate the feasibility of high-sensitivity near-infrared fluorescence imaging with indocyanine green for intraoperative identification of hepatocellular carcinoma in dogs. METHODS Twelve hepatic nodules were surgically resected from six dogs. In each dog, 0 · 5 mg/kg indocyanine green was intravenously injected for 12 to 18 hours preoperatively. The hepatic nodules were investigated under laparotomy using a near-infrared fluorescence imaging light camera system prior to resection. Resected nodules were histopathologically diagnosed and their fluorescence images were evaluated. RESULTS Of the 12 hepatic nodules, 6 were diagnosed as hepatocellular carcinoma and 6 as nodular hyperplasia. Indocyanine green-fluorescence was observed in four large hepatocellular carcinoma nodules and one case of nodular hyperplasia, whereas it was absent in the remaining nodules. The sensitivity and positive predictive values of indocyanine green fluorescent imaging for hepatocellular carcinoma was 71 · 4 and 80 · 0%, respectively. Complete resection of the hepatic masses was achieved in all dogs. CLINICAL SIGNIFICANCE Near-infrared fluorescence imaging may be feasible for intraoperative mapping of hepatocellular carcinomas in hepatic lobes and may help increase the chance of complete resection of hepatocellular carcinoma in dogs.

Effects of Cryopreservation on the Cell Viability, Proliferative Capacity and Neuronal Differentiation Potential of Canine Bone Marrow Stromal Cells

ABSTRACT We investigated the cell viability, proliferative capacity and neuronal differentiation potential of canine bone marrow stromal cells (BMSCs) after cryopreservation. BMSCs were cryopreserved using cryoprotectant solutions with 10% DMSO and 10% FBS (DF group) or without DMSO and FBS (DF-free group); fresh BMSCs were used as a control. The cell viability and proliferative capacity of BMSCs were similar in the DF-free and control groups, while those in the DF group were lower. In all groups, BMSCs differentiated into neuron-like cells that stained positive against neuron markers, and the mRNA expression levels of neuron markers increased after neuronal induction. In conclusion, cryopreservation with DF-free cryoprotectant solution did not diminish the cell viability, proliferative capacity or neuronal differentiation potential of canine BMSCs.

Noninvasive estimation of cardiac systolic function using continuous‐wave Doppler echocardiography in dogs with experimental mitral regurgitation

OBJECTIVE To evaluate the feasibility of noninvasive estimation of cardiac systolic function using transthoracic continuous-wave Doppler echocardiography in dogs with mitral regurgitation. PROCEDURE Seven mongrel dogs with experimental mitral regurgitation were used. Left ventriculography and measurement of pulmonary capillary wedge pressure were performed under inhalational anaesthesia. A micromanometer-tipped catheter was placed into the left ventricle and transthoracic echocardiography was carried out. The peak rate of left ventricular pressure rise (peak dP/dt) was derived simultaneously by continuous-wave Doppler and manometer measurements. The Doppler-derived dP/dt was compared with the catheter-measured peak dP/dt in the dogs. RESULTS Classification of the severity of mitral regurgitation in the dogs was as follows: 1+, 2 dogs; 2+, 1 dog; 3+, 2 dogs; 4+, 1 dog; and not examined, 1 dog. We were able to derive dP/dt from the transthoracic continuous-wave Doppler echocardiography in all dogs. Doppler-derived dP/dt had a significant correlation with the catheter-measured peak dP/dt (r = 0.90, P < 0.0001). CONCLUSION It was demonstrated that transthoracic continuous-wave Doppler echocardiography is a feasible method of noninvasive estimation of cardiac systolic function in dogs with experimental mitral regurgitation and may have clinical usefulness in canine patients with spontaneous mitral regurgitation.

Successful Resolution of Esophageal Granulomas in a Dog Infected with Spirocerca lupi

ABSTRACT A 13-year-old female Labrador Retriever presented with chronic regurgitation. Radiography and computed tomography (CT) revealed nodules in the caudal esophagus. Upper gastrointestinal endoscopy followed by histopathological examination revealed esophageal granulomas caused by Spirocerca lupi. The infection was treated with milbemycin oxime. The therapy was successful, and a remarkable reduction in granulomas was observed. This case report describes the diagnostic imaging findings and treatment outcome of a dog with S. lupi infection in Japan.

Preoperative Differential Diagnosis of Canine Adrenal Tumors Using Triple-Phase Helical Computed Tomography.

OBJECTIVE To characterize the computed tomography (CT) findings for canine adrenal tumors, including cortical adenoma, cortical adenocarcinoma, and pheochromocytoma, and to evaluate the feasibility and usefulness of preoperative triple-phase helical CT for differentiation of tumor types and surgical planning. STUDY DESIGN Retrospective study. ANIMALS Dogs with adrenal tumors (n=36). METHODS All dogs underwent triple-phase helical CT, followed by adrenalectomy and histopathological diagnosis of the resected mass. Precontrast images, arterial, venous, and delayed phase images were obtained. In all cases, morphological characteristics and CT values and calculations, including the percentage enhancement washout ratio, relative percentage washout, enhancement washin, and enhancement washout, were analyzed and compared among the tumor types. RESULTS Of the 36 dogs with adrenal masses, cortical adenocarcinoma was most commonly diagnosed (16 dogs), followed by pheochromocytoma (13 dogs), and cortical adenoma (7 dogs). The precontrast minimum CT value and enhancement washout between venous and delayed phases in the cortical adenoma were significantly higher than those in the cortical adenocarcinoma. The maximum CT values of the precontrast image and arterial and venous phases, the enhancement washin and washouts, percentage enhancement washout ratio, and relative percentage washout in the pheochromocytomas were significantly higher than those in cortical adenocarcinoma. CONCLUSION The differential diagnosis of canine adrenal tumors was feasible based on triple-phase CT findings, including morphological features, CT values, and intratumoral contrast attenuation. Preoperative diagnosis using triple-phase helical CT may be useful for surgical planning in dogs with adrenal tumors.