Takefumi Ishikawa

Identification of Cancer Stem Cells Derived From a Canine Lung Adenocarcinoma Cell Line

Accumulating evidence supporting the cancer stem cell (CSC) hypothesis is based on the finding that tumors contain a small population of self-renewing cells that generate differentiated progeny and thereby contribute to tumor heterogeneity. CSCs are reported to exist in several human cancers, yet only a few reports demonstrate the existence of CSCs in primary lung cancer in dogs. In this study, the authors established a cancer cell line derived from a canine primary lung adenocarcinoma and identified a side population (SP) of cells that displayed drug-resistant features. To confirm the characteristics of these SP cells, the authors investigated the tumorigenicity of the cells in vivo by using a nude mouse xenograft model. Only 100 SP cells were able to give rise to new tumors, giving a 10-fold enrichment over the main population (MP) of cells, suggesting that these cells have the cancer-initiating ability of CSCs. Further studies characterizing CSCs in canine lung adenocarcinoma might contribute to the elucidation of the mechanisms of tumorigenesis and to the establishment of novel therapeutic strategies.

Prothrombotic and Inflammatory Effects of Intravenous Administration of Human Immunoglobulin G in Dogs

BACKGROUND Intravenous administration of human immunoglobulin G (hIVIgG) has been suggested to potentiate thromboembolism in dogs, but supportive scientific reports are lacking. OBJECTIVES To determine if hIVIgG therapy promotes hypercoagulability and inflammation in dogs. ANIMALS Twelve healthy Beagle dogs. METHODS Prospective, experimental trial. An hIVIgG/saline solution was infused IV at 1 g/kg BW over 8 hours to 6 dogs, and physiological saline was infused to the other 6 dogs. Blood samples were drawn before, during, and after infusion for serial measurement of indicators of coagulation and inflammation. Data were analyzed by 2-way repeated measures analysis of variance. RESULTS Dogs administered hIVIgG developed mildly decreased blood platelet concentrations without thrombocytopenia (median, 200 x 10(3)/microL; range, 150-302 x 10(3)/microL; P < .01), leukopenia (median, 3.5 x 10(3)/microL; range, 20-62 x 10(3)/microL; P < .001), and mildly increased plasma total protein concentrations (median, 6.3 g/dL; range, 5.6-6.7 g/dL; P < .001). Administration of hIVIgG was also associated with increases in fibrin/fibrinogen degradation products in all dogs (either 5 microg/mL or 10 microg/dL), thrombin-antithrombin III complexes (median, 7.2 ng/mL; range, 4.9-14.2 ng/mL; P < .001), and C-reactive protein concentrations (median, 2.5 mg/dL; range, 0.5-4.3 mg/dL; P < .01). CONCLUSION AND CLINICAL IMPORTANCE Administration of hIVIgG to dogs promotes hypercoagulability and an inflammatory state. This should be further evaluated and considered when using hIVIgG in dogs with IMHA or other prothrombotic conditions.

Alpha-fetoprotein in serum and tumor tissues in dogs with hepatocellular carcinoma

Serum alpha-fetoprotein (AFP) concentrations were measured before and after surgical removal of tumor masses in four dogs with hepatocellular carcinoma (HCC). Localization of AFP was also examined immunohistochemically in tumor tissues. In three cases, the serum AFP concentration was 10 to 20 times higher than that of normal dogs. One to two months after surgery, the serum AFP concentration had decreased to normal range. AFP was localized in the tumor tissues in these three cases. One case, which had a low serum AFP, did not show AFP localization in tumor tissue.

Canine bone marrow cells differentiate into hepatocyte-like cells and placental hydrolysate is a potential inducer

Hepatocyte growth factor (HGF) can stimulate human and rat bone marrow (BM) cells to differentiate into hepatocytes. A human placental hydrolysate (hPH) stimulates proliferation of hepatocytes, but its role as a potential inducer of BM cells to form hepatocytes is unclear. To determine if canine BM cells stimulated with HGF or hPH differentiate into hepatocyte-like cells, BM cells were cultured with HGF or hPH. The cultured cells underwent morphological examination, expression of albumin and cytokeratin 18 (CK18), hepatic function tests including uptake of low-density lipoprotein (LDL) and cytochrome P (CYP) 450 activity. Albumin mRNA and protein expression of albumin and CK18 proteins were detected in cultures with HGF and hPH. Furthermore, these cells demonstrated LDL uptake and CYP450 activity. These results indicate that canine BM cells can differentiate into hepatocyte-like cells when stimulated by both HGF and that hPH may be an effective inducer of hepatic differentiation.

Partial Anomalous Pulmonary Venous Connection in 2 Miniature Schnauzers

A 3-year-old, 6.0 kg, intact female Miniature Schnauzer was presented to Azabu University for evaluation of right heart enlargement, incidentally noticed on survey thoracic radiographs. The dog was asymptomatic and no abnormalities were identified on physical examination. Radiographic evaluation of the thorax indicated right heart enlargement (vertebral heart score, 12.1). Two-dimensional echocardiography disclosed right atrial and right ventricular dilatation (Fig 1). An abnormal vascular structure connected to the right atrium at the heart base was observed on color Doppler echocardiography (Fig 2). No other structural heart disease or conditions that could result in right heart dilatation (eg, pulmonary hypertension, atrial septal defect [ASD], tricuspid valve regurgitation) were found. A complete blood count (CBC) and serum biochemistry profile were within normal limits. D-dimer concentration (reference range, <0.2 lg/mL) was normal. Computed tomography angiography (CTA) and cardiac catheterization were performed under general anesthesia, maintained by fentanyl constant rate infusion and isoflurane inhalation, to determine a definitive diagnosis. A 4 Fr multipurpose catheter was introduced from the right jugular vein through a catheter introducer placed by the Seldinger technique. Oxygen saturation (SaO2) of each site within the heart was measured while breathing room air. Mean SaO2 of the cranial and caudal vena cava was 55.9%. SaO2 in the right atrium varied from 81.8 to 99.2%, depending on the location of the catheter tip. SaO2 at the right ventricle and pulmonary artery were 77.0 and 83.2%, respectively. Mean right atrial and pulmonary pressures were 1 and 11 mmHg, respectively. The dog was placed in dorsal recumbency on a clinical 16-multi-detector-row computed tomography scanner. Iodinated contrast medium (2 mg/kg) was rapidly injected via the cephalic vein. Repetitive transverse plane cine scans (120 kV, 99 mAs, 0.625 mm slice thickness, 0.6 s tube rotation time, 0.938 helical pitch) were acquired over the heart. Images were transferred to an image software system for further evaluation. Acquired images were analyzed using multiplanar reconstruction and volume rendering, and it was determined that the pulmonary vein of the right cranial lung lobe was connected to the right atrium. Therefore, a definitive diagnosis of partial anomalous pulmonary venous connection (PAPVC) was made (Fig 3).

Effects of Prostaglandin E1 on the Preparation of Platelet Concentrates in Dogs

BACKGROUND Platelet concentrates (PC) are prepared by centrifugation of platelet-rich plasma (PRP) that is prepared by centrifugation of whole blood. The resuspension of the platelet pellet during PC preparation from dogs is difficult because of platelet activation induced by centrifugation. OBJECTIVES To investigate the efficacy of adding prostaglandin E(1) (PGE(1) ) to prevent platelet activation during PC preparation from dogs. ANIMALS Fifteen healthy Beagle dogs. METHODS Prospective, experimental trial: PGE(1) was added to PRP before the high-speed centrifugation during PC preparation. To estimate the effect of this addition, we assessed the platelet aggregability before transfusion, the survival of the platelets after transfusion, and the platelet reactivity after transfusion, which is estimated by the P-selectin expression of the platelets when stimulated by thrombin. RESULTS The difficulty associated with platelet resuspension was resolved by PGE(1.) PGE(1) strongly inhibited platelet aggregation induced by collagen and ADP; however, it recovered after the platelets were resuspended in plasma without PGE(1) (mean aggregation ratio; collagen: 10.00-80.80%, ADP: 8.20-53.60%). Survival of the platelets after transfusion was not affected by PGE(1) (mean 8.04 and 7.56 days, without and with PGE(1) ), and thrombin-induced P-selectin expression after transfusion in PGE(1) -treated PC was also well maintained (mean positive ratio 53.7 and 47.9%, before and 24 hours after transfusion). CONCLUSIONS AND CLINICAL IMPORTANCE The addition of PGE(1) in PRP before the centrifugation of PRP can improve the preparation efficiency of PC from dogs, while maintaining the therapeutic efficacy of the platelets.