Novel variations and loss of heterozygosity of BRCA2 identified in a dog with mammary tumors

OBJECTIVE: To establish novel polymorphic markers for analysis of loss of heterozygosity (LOH), so as to study the possible involvement of BRCA2 in mammary tumors obtained from dogs. SAMPLE POPULATION: Blood samples, mammary gland specimens, or mammary tumors from 3 tumor-bearing dogs and 10 tumor-free dogs. PROCEDURES: Nucleotide sequence analysis was performed with a DNA autosequencer. Loss of heterozygosity analysis was performed for markers established in the present study. The expression level of canine BRCA2 was quantified by real-time PCR analysis. RESULTS: 3 novel microsatellite markers with high heterozygosity rates (> 50%) were established, and the previously reported marker for canine BRCA2 gene locus was improved. These markers were used for the analysis of DNA from formalin-fixed and paraffin-embedded samples. By use of these markers, LOH in canine BRCA2 was identified as a result of recombination. In mammary tumor DNA that corresponded to the LOH-positive dog, the level of canine BRCA2 expression was decreased compared with that of nonneoplastic mammary gland tissue; the open reading frame contained 4 missense variations, 1 insertion variation, and 1 silent variation, some of which were localized to functional domains. CONCLUSIONS AND CLINICAL RELEVANCE: 3 novel polymorphic markers were developed for LOH analysis of canine BRCA2 and identified a dog with LOH with some variations in the functional domains. These markers could be useful for assessing the relevance of BRCA2 variation in mammary tumors of dogs.     

Successful use of prednisolone and radiation therapy in a dog with intracranial histiocytic sarcoma

The ideal treatment for intracranial histiocytic sarcoma (HS) remains unclear. Herein, we report a case of intracranial HS that was successfully treated using prednisolone and radiation therapy. The patient was a 9-year-old spayed female Pembroke Welsh Corgi that presented with epileptic seizures. Magnetic resonance imaging revealed a contrast-enhancing mass adjacent to the right piriform lobe. Prednisolone administration (1 mg/kg/day) decreased the lesion size. Additional palliative radiation therapy (total dose, 37 Gy) resulted in complete disappearance of the lesion. However, on day 164, the dog’s neurological signs deteriorated, and she was euthanized. Necropsy revealed an intracranial metastasis of HS via the cerebrospinal fluid without any extracranial metastasis. Nonetheless, combined prednisolone and radiation therapy might be effective in treating intracranial HS.     

A pre-harvest method for predicting understory light levels created by artificial canopy gaps

A new method for predicting understory light levels around an artificial canopy gap prior to the selective cutting is presented. In two even-aged coppice stands, hemispherical photographs were taken at a height of 1.3 m, and crowns of each canopy stem on the photographs were identified by field observations. To predict light levels after cutting, we used computerized images of the hemispherical photographs, in which we digitally erased all crowns to be cut (named ‘erasing method’). Light levels were calculated by an analyzing program; HEMIPHOT. Light levels predicted by the ‘erasing method’ were compared with that calculated by the ordinary photograph-analyzing method using photographs taken after the actual cutting. The correlations between the two methods were highly significant in the two stands. The ‘erasing method’ is eminently effective for the prediction of understory light levels in an artificial canopy gap.     

Dynamic left ventricular outflow tract obstruction secondary to hypovolemia in a German Shepard dog with splenic hemangiosarcoma

Dynamic left ventricular outflow tract obstruction (DLVOTO) is a common condition in cats and humans. In this case report, a dog is described with DLVOTO secondary to severe intra-abdominal hemorrhage caused by a hemangiosarcoma. The dog was a 9-year-old, 35.7-kg, spayed female German Shepard dog that presented with a history of tachypnea and collapse. A Levine II/VI systolic murmur was present at the heart base. Abdominal ultrasonography revealed a splenic mass and a large amount of ascites. Echocardiography showed a reduced left ventricular diameter and an increased aortic velocity caused by systolic anterior motion (SAM) of the mitral valve apparatus. The heart murmur and the SAM were resolved after treatment including a splenectomy and a blood transfusion.     

马尾松早期促进开花结实方法的研究

１９９７年在京山县太子山林场５年生马尾松无性系种子园内,用三种生长调节剂Ｃ．Ｃ．Ｃ．ＧＡ,ＢＡＰ分施用量、处理时期、处理次数三个因素进行叶面喷洒实验。结果表明ＢＡＰ有促进开花的作用。１９９８年在中心苗圃３年生嫁接苗实验区,利用１９９７年对促进开花结实有效果的ＢＡＰ进一步分时期、处理次数二因素进行了实验,结果表明ＢＡＰ对马尾松苗期开花也有明显的促进作用。两年的实验结果表明,ＢＡＰ能促进马尾松早期开花,这对缩短马尾松育种周期,加速新品种的开发具有非常现实的意义。     

Endogenous 5-HT outflow from chicken aorta by 5-HT uptake inhibitors and amphetamine derivatives

Chemoreceptor cells aggregating in clusters in the chicken thoracic aorta contain 5-hydroxytryptamine (5-HT) and have voltage-dependent ion channels and nicotinic acetylcholine receptors, which are characteristics typically associated with neurons. The aim of the present study was to investigate the effects of 5-HT uptake inhibitors, fluvoxamine, fluoxetine and clomipramine (CLM), and amphetamine derivatives, p-chloroamphetamine (PCA) and methamphetamine (MET), on endogenous 5-HT outflow from the isolated chick thoracic aorta in vitro. 5-HT was measured by using a HPLC system with electrochemical detection. The amphetamine derivatives and 5-HT uptake inhibitors caused concentration-dependent increases in endogenous 5-HT outflow. PCA was about ten times more effective in eliciting 5-HT outflow than MET. The 5-HT uptake inhibitors examined had similar potency for 5-HT outflow. PCA and CLM increased 5-HT outflow in a temperature-dependent manner. The outflow of 5-HT induced by PCA or 5-HT uptake inhibitors was independent of extracellular Ca²⁺ concentration. The 5-HT outflow induced by CLM, but not that by PCA, was dependent on the extracellular NaCl concentration. These results suggest that the 5-HT uptake system of 5-HT-containing chemoreceptor cells in the chicken thoracic aorta has characteristics similar to those of 5-HT-containing neurons in the mammalian central nervous system (CNS).     

Isolation of avian bornaviruses from psittacine birds using QT6 quail cells in Japan

Avian bornaviruses (ABVs) were recently discovered as the causative agents of proventricular dilatation disease (PDD). Although molecular epidemiological studies revealed that ABVs exist in Japan, no Japanese isolate has been reported thus far. In this study, we isolated four strains of Psittaciform 1 bornavirus from psittacine birds affected by PDD using QT6 quail cells. To our knowledge, this is the first report to isolate ABVs in Japan and to show that QT6 cells are available for ABV isolation. These isolates and QT6 cells would be powerful tools for elucidating the fundamental biology and pathogenicity of ABVs.