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Repeatable head immobilization is important for minimizing positioning error during radiation therapy for veterinary patients with head neoplasms. The purpose of this retrospective cross-sectional study was to describe a novel technique for head immobilization (Device II) and compare this technique with a previously described technique (Device I). Device II provided additional support by incorporating three teeth (vs. two teeth with Device I). Between 2011 and 2013, both devices were applied in clinically affected cats (Device I, n = 17; Device II, n = 11) and dogs (Device I, n = 85; Device II, n = 22) of various breeds and sizes. The following data were recorded for each included patient: variability in the angle of the skull (roll, yaw, and pitch), coordinates of the isocenter, and distance from the reference mark to the tumor. Devices I and II differed for skull angle variability during the treatment of dogs (roll, P = 0.0007; yaw, P = 0.0018; pitch, P = 0.0384) and for yaw of during the treatment of cats (P < 0.0001). In each case, Device II was superior to Device I. The distance from the reference mark to the center of the tumor was significantly decreased for Device II vs. Device I (dogs, P < 0.0001; cats, P = 0.0002). Device II also provided more accurate coordinates for the isocenter. Authors recommend the use of, Device II for future clinical patients.

In this report, we describe a case of lipomatosis in the left parotid gland of an eight-year-old female Shetland sheepdog and review the relevant literature. Preoperative diagnosis of lipomatosis with Tru-Cut biopsy presented difficulties in this case. The postoperative diagnosis was based on the gross appearance of a non-infiltrative, circumscribed swollen mass in the parotid gland and the histological appearance of normal adipocytes that infiltrated into the parotid gland without involving surrounding structures. Additionally, flotation of the whole parotid gland in formalin solution together with well-maintained residual lobulation and a well-maintained configuration on the cut surface were identified as subsidiary characteristics of lipomatosis of the parotid gland. Lipomatosis of the parotid gland has been rarely reported in dogs and is a poorly understood condition.

Cancer stem-like cells (CSCs)/cancer-initiating cells (CICs) are a small subpopulation of cancer cells that are responsible for the initiation, recurrence and metastasis of cancer. We previously demonstrated that, using the Hoechst 33342 dye-based side population technique, CSCs/CICs in canine lung adenocarcinoma cell line exist. In this study, as CSCs/CICs are known to form spheres in anchorage-independent environment in vitro, we evaluated the stemness of spheroid cells derived from canine lung adenocarcinoma and osteosarcoma cells by expression of stemness markers, and investigated radioresistance. Spheroid cells showed greater expression of stemness markers Oct-4 and CD133 gene than those of adherent-cultured cells. In nude mouse xenograft models, spheroid cells showed higher tumourigenic ability than adherent-cultured cells. In addition, spheroid cells showed significantly resistant against radioactivity as compared with adherent-cultured cells. These results suggest that spheroid cells could possess stemness and provide a CSCs/CICs research tool to investigate CSCs/CICs of canine tumour cells.

Tumor-initiating cells (TICs) or cancer stem cells (CSCs), a small subset of tumor cells, are involved in tumor initiation, progression, recurrence and metastasis. In human hepatocellular carcinoma (HCC), TICs are enriched with cell surface markers and have the ability to self-renew and differentiate tumors at a high frequency. We established a canine HCC cell line, HCC930599, and analyzed it for stem and progenitor cell marker expression using flow cytometry. HCC930599 showed high CD44 and CD29, moderate CD90, and low CD133, CD34, CD24, CD117, and CD13 expression. CD90(+)CD44(+) and CD90(-)CD44(+) cells were characterized using the in vitro sphere assay and an in vivo transplant model. CD90(+)CD44(+) cells acquired enhanced self-renewal capacity, proliferative activity and tumourigenicity compared with CD90(-)CD44(+) cells, suggesting that TICs exist in the HCC930599 cell line and that CD90 is a marker for enriched TICs. Understanding TIC characteristics may help elucidate hepatic carcinogenesis and HCC therapy development.

We evaluated the efficacy of weekly bathing in reducing methicillin-resistant Staphylococcus (MRS) colonization in canine allergic dermatitis in a pilot clinical trial. Six dogs with allergic dermatitis controlled by prescription medications were treated with weekly bathing for 1 month. The Canine Atopic Dermatitis Extent and Severity Index version 3 (CADESI-03) and pruritus scores and frequency of mecA-positive Staphylococcus spp. isolated from three body sites between weeks 0 and 4 were compared. There was no significant difference in CADESI-03 scores with bathing, whereas the pruritus scores were significantly reduced (p0.05). Furthermore, MRS frequency was decreased in four of the six dogs (p0.05). In conclusion, weekly bathing should be considered for reducing MRS colonization in canine allergic dermatitis.

ミニチュア・ダックスフント，オス，12歳が，鼻稜部，体幹部，足根関節部の皮膚病変，腹部膨満と多飲多尿（192 ml/kg/day）を呈して来院した。ACTH刺激試験では，投与前のコルチゾール値が12.1 μg/dl，投与1時間後が68.4 μg/dlであった。下垂体依存性副腎皮質機能亢進症（PDH）と診断し，小分割照射（毎週1回［6 Gy］，合計3回［18Gy］）を実施した。飲水量は徐々に減少し，約1年後には85 ml/kg/dayとなり，被毛も改善した。ACTH刺激試験では，投与前が6.3 μg/dl，投与1時間後が36.1 μg/dlであった。ACTH試験では依然高値であったが，PDHに対する低線量小分割照射は，臨床症状の改善には有効である可能性が示唆された。

An 8-year-old female Golden Retriever had an oral mass and lameness. Multiple osteolysis of the systemic skeleton without monoclonal gammopathy was shown on electrophoresis of serum and urine samples. Cytological and histopathological examinations of the oral mass revealed atypical polymorphic cells similar to myeloid cells, and bone marrow aspiration indicated that these abnormal cells also might have invaded the bone marrow. These cells were negative to peroxidase and non-specific esterase staining, and clonal expansion of B lymphocytes could be detected by polymerase chain reaction (PCR) assay for antigen receptor gene rearrangement. The case was diagnosed as atypical lymphoma and treated by multi-drug chemotherapy. On the 142nd day after the first admission, the case had remission and the oral mass and multiple osteolysis were improved.

肺炎を併発した肺原発の高分化型気管支肺胞腺癌 (T2 Nx M0) の8歳のパグ, 雌, 9kgに対して副葉以外の右側肺葉切除を行い, さらに術後に低用量のカルボプラチン (120mg/m2, iv, 3～4週ごと) を1年2ヵ月間投与し, 19ヵ月の生存期間を得た。犬の肺癌は予後不良な腫瘍の1つではあるが, 高分化型の肺癌の場合には積極的な外科療法によって長期生存できる場合があることが明らかとなった。また, その効果は明らかではないが, 低用量のカルボプラチン療法は副作用も少なく, 検討に値する治療法と考えられた。

Repeatable head immobilization is important for minimizing positioning error during radiation therapy for veterinary patients with head neoplasms. The purpose of this retrospective cross-sectional study was to describe a novel technique for head immobilization (Device II) and compare this technique with a previously described technique (Device I). Device II provided additional support by incorporating three teeth (vs. two teeth with Device I). Between 2011 and 2013, both devices were applied in clinically affected cats (Device I, n = 17; Device II, n = 11) and dogs (Device I, n = 85; Device II, n = 22) of various breeds and sizes. The following data were recorded for each included patient: variability in the angle of the skull (roll, yaw, and pitch), coordinates of the isocenter, and distance from the reference mark to the tumor. Devices I and II differed for skull angle variability during the treatment of dogs (roll, P = 0.0007; yaw, P = 0.0018; pitch, P = 0.0384) and for yaw of during the treatment of cats (P0.0001). In each case, Device II was superior to Device I. The distance from the reference mark to the center of the tumor was significantly decreased for Device II vs. Device I (dogs, P0.0001; cats, P = 0.0002). Device II also provided more accurate coordinates for the isocenter. Authors recommend the use of, Device II for future clinical patients.

A 10-year-old castrated male Golden retriever, weighing 36.3 kg was referred for evaluation owing to a decline in general condition. Findings from the complete blood count revealed a marked lymphocytosis (113 000/µℓ). Examination of Wright-Giemsa-stained films of peripheral blood revealed the presence of large granular lymphocytes (LGL). Seventy-two per cent (81 360/µℓ) of the lymphocytes were found to be 12-17 µm in diameter, containing nuclei with mature clumped chromatin and abundant lightly basophilic cytoplasm with a variable number of fine azurophilic granules. Based on these findings this case was diagnosed as LGL leukaemia. As a result of multiple-agent chemotherapy, the markedly elevated levels of lymphocytes gradually decreased to 7500/µℓ on day 122 and the patient maintained a good quality of life for the following 3 months. However, on around day 237, a soft, raised, bosselated mass on the labial region was noted. The dog was diagnosed as having histiocytic sarcoma based on cytological and histological examination of the mass. Shortly after diagnosis, the dog developed sudden onset of central nervous system signs and died on day 270. A common outcome of canine LGL is the development of acute blast crisis or lymphoma. However, this case was notable for complication with histiocytic sarcoma from another origin.

Diffusion into the ovarian artery of prostaglandin F2α (PGF2α) infused into the uterine vein of a cow was studied to demonstrate the existence of the so-called “utero-ovarian counter current mechanism” which was responsible for luteal regression in sheep and guinea pigs. In the first part of Exp. 1, PGF2α was infused in the uterine vein through an indwelling cannula. The plasma PGF2α concentration in the ovarian artery was increased to more than 300 times that in the jugular vein 1 min after the start of injection. In the second part which was begun 15 min after the completion of final blood sampling in the first part the plasma PGF2α concentration in the ovarian artery was increased to 2.7 times that in the jugular vein one minute after the start of injection. Though the difference between the PGF2α concentrations in the arterial and venous blood was smaller in the second part of Exp. 1, these results clearly show the existence of the counter current mechanism in the cow. In Experiment 2, PGF2α (15 mg in 3 ml solvent) was injected into the gluteal muscle 30 min after the completion of the final blood sampling to examine the changes in plasma concentrations of PGF2α in the ovarian artery, jugular vein and uterine vein after systemic injection of PGF2α. The PGF2α concentration in uterine venous blood prior to intra-gluteal injection was extremely high (60000 pg/ml). The concentrations in the ovarian artery and jugular vein were slightly increased with time after injection. Although the value was somewhat higher in the ovarian artery, almost similar patterns of change were seen in both the ovarian artery and the jugular vein.

Metastasis of a malignant mixed tumor occurring in the abdominal wall was found in the liver of a 15-year-old dog male. The vascular system of the metastatic lesion was examined by scanning electron microscopy using the resin cast technique. The existence of arterio-portal shunts was confirmed. The shunts arose from branches of the hepatic artery in retrograde or vertical direction and merged into the capillary plexus replacing the branches of the portal vein, forming the so-called thread and streaks vessel in the field of hepatic angiography. Vessels with the thread and streaks appearance histologically represented tumor emboli growing in the lumen of a large branch of the portal vein. There were capillaries running inside along the portal vein and forming a longitudinal dense network in the vein. Branches of the hepatic artery feeding the metastatic lesion were well developed and formed a coarse arterial network in their course. Tributaries of the hepatic vein from the lesion were also well developed.

A simple method of lymphography of the thoracic duct was investigated. Using three female beagles, contrast media were administered rectally, vaginally and into the perianal tissue. The administration sites were gently massaged, and imaging was carried out at constant intervals using computed tomography and radiograph. Moreover, Indian ink was administered into the rectum mucous membrane in dogs for proof of this method of lymphography, and the lymph drainage routes were observed. The investigation showed that clear computed tomography and radiographic contrast images of the thoracic duct were obtained by subcutaneous and submucosa injection of angiography contrast medium and 3D processing of these images revealed the three-dimensional positions and course of the thoracic duct and cisterna chyli.

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.

The object of this study was to evaluate hypofractionated multiportal field and two-portion (rostral and caudal portions divided by the eyelid) radiation therapy for canine nasal tumors. Sixty-three dogs underwent multiportal hypofractionated radiation therapy. The radiation field was divided into rostral and caudal portions by the eyelid. Treatments were performed four times for 57 dogs. The median irradiation dose/fraction was 8 Gy (range, 5-10 Gy); the median total dose was 32 Gy (10-40 Gy). Improvement of clinical symptoms was achieved in 53 (84.1%) of 63 cases. Median survival time was 197 days (range, 2-1,080 days). Median survival times with and without destruction of the cribriform plate before radiotherapy were 163 and 219 days, respectively. There was no significant difference between them. No other factors were related to survival according to a univariate analysis. All radiation side effects, except one, were grade I according to the VRTOG classification. It was not necessary to treat any dogs for skin side effects. One dog (1.6%) developed an oronasal fistula 1 year after completion of radiation therapy. This radiation protocol may be useful in reducing radiation side effects in dogs with cribriform plate destruction.

This retrospective study examined 8 cases of pathologically diagnosed insulinoma. Surgical intervention was performed in all cases, once insulinoma was suspected based on unique clinical signs, hypoglycemia, and/or findings on ultrasonography. At the same time, insulin and glucose concentrations were measured before surgery for clinical diagnosis. Although all cases displayed typical clinical signs and hypoglycemia, insulin concentrations varied with 3 of 8 cases showing levels within the reference interval. In addition, to confirm the reliability of measured values, we submitted serum samples from 4 cases to two commercial veterinary laboratories. Results differed considerably between laboratories, with no apparent correlations between the two. In Laboratory A, 3 of 4 cases were above the reference interval, and 1 case was in the middle of the reference interval. Conversely, in Laboratory B, 3 of 4 cases were above the reference interval, and 1 case was below the reference interval. Split decisions regarding the diagnosis of insulinoma were seen for 2 of the 4 cases.

A West Highland White Terrier was brought to our veterinary hospital with the chief complaints of seizures and staggering gait. When cephalic Magnetic resonance imaging (MRI) was performed, a symmetrical lesion was found in the basal ganglion. Suspecting a metabolic disorder, an abdominal ultrasonography was performed, and a tumor was found in the pancreas. The pancreatic tumor was surgically removed based on suspicion that it had induced the brain damage. The resected tumor was histopathologically diagnosed as an insulinoma. After removal, recurrence of neurological symptoms was not observed. MRI 3 months post-operation showed a reduction in the size of the brain lesion. Consequently, the tumor was thought to have induced the lesion in the basal ganglion, and this was verified by MRI. This case was considered to be extremely rare.

We examined the utility of a bite block-type head immobilization device, hereafter referred to as "head immobilization device", in order to improve the ease of immobilization and accuracy when performing radiotherapy for cranial tumors in animals. The head immobilization apparatus was a rectangular-shaped bite block-type device. We examined 55 cases in 46 dogs that underwent head CT scans between June 2005 and May 2006. The head immobilization device was used for 26 cases (immobilization group) and was not used for 29 cases (control group). Head stability was maintained in the control group by placing a towel under the head. We measured the angle of rotation of the xy, yz and xz planes for each group. The angles of rotation of the xy plane for the control and immobilization groups were 3.69 +/- 2.28 (mean +/- SD) and 1.39 +/- 1.50, respectively. The t-test demonstrated that the difference was statistically significant (p0.001). These results indicate that there was reduced tilting to the left or right. We conclude that use of this head immobilization device was extremely easy and that it improved the accuracy of radiotherapy for cranial tumors in dogs and cats.

With radiography and ultrasound, reversed positioning of the fundus ventriculi and pylorus, a duodenum running on the left side, transposition of the kidneys, and normal thoracic organs were found in a 5-month-old miniature dachshund that presented with anorexia and weight loss. The case was diagnosed as partial heterotaxia. Gross observation revealed partial heterotaxia, polysplenia, abnormal lobulation of the liver, and absence of the greater omentum. These findings were consistent with those observed in asplenia-polysplenia syndrome in humans.

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.

A hepatoblastoma was found in a 13-year-old female Maltese dog. Histologically, the tumor showed a wide trabecular pattern and was frequently accompanied with vascular lake formation. Tumor cells were positive for cytokeratin and neuron specific enolase, but negative for chromogranin. Electronmicroscopically, tumor cells were accompanied with continuous basement membrane and had poorly developed desmosomes. Sinusoidal endothelia had fenestration and were surrounded by myofibroblast-like cells. To the best of our knowledge, this paper is the first report of morphological studies on canine hepatoblastoma.

Hepatic nodular fibrosis occurred in an 8-year-old male Papillon dog. Fibrous nodules, consisting of broad bands of collagen fibers, spindle cells, and lipofuscin-laden foamy macrophages, were well-circumscribed and frequently linked up with the portal areas. Because the spindle cells were positive for desmin and/or alpha-smooth muscle actin, they might be Ito cells or myofibroblasts. These results suggest that both the spindle cells and macrophages play an important role in the pathogenesis of hepatic nodular fibrosis, which might arise from the portal area.