Changes in extracellular neurotransmitters in the cerebrum of familial idiopathic epileptic shetland sheepdogs using an intracerebral microdialysis technique and immunohistochemical study for glutamate metabolism

Intracerebral microdialysis combined with electroencephalographic recordings was performed on 4 dogs of a familial idiopathic epileptic Shetland sheepdog colony to identify the kinds of neurotransmitters responsible for seizure activity. Immunohistochemistry using glutamate (Glu), glutamate transporter (GLT-1 and GLAST), and glutamine synthetase (GS) antibodies was also carried out on the cerebrum of four familial dogs that died of status epilepticus (SE). High values for extracellular levels of Glu and aspartate (ASP) were detected in association with an increased number of spikes and sharp waves during hyperventilation in 3 of 4 the familial epileptic dogs. The values of other amino acids analyzed were not altered in any of the familial epileptic dogs. Immunohistochemically, Glu-positive granules were occasionally found in the perineuronal spaces of the cerebral cortex in 3 of the familial epileptic dogs that died of SE. Immunostains for GLT-1 antibody predominantly decreased in the cerebral cortex and lateral nucleus of the thalamus in all the dogs that died of SE, whereas there were no differences detected in immunolabellings for GLAST and GS antibodies between familial epileptic dogs and controls. These results suggest that an extracellular release of both Glu and Asp may play an important role in the occurrence of seizure activity in this epileptic colony, and that a decreased expression of astrocytic GLT-1 may be related to development of SE.     

Myofiber expression of class I major histocompatibility complex accompanied by CD8+ T-cell-associated myofiber injury in a case of canine polymyositis

A 7-year-old female Labrador Retriever dog showed extreme muscular weakness, muscle wasting, dysbasia, and mild dysphagia. An elevated value of creatine kinase (335 IU/liter) in the serum was detected. Electromyographic findings included increased insertional activity, fibrillation potentials, and bizarre high-frequency repetitive potentials. Histopathologic examination of skeletal muscles revealed myofiber necrosis and phagocytosis, regeneration of myofibers, and perivascular, perimysial, and endomysial infiltrations of lymphocytes, macrophages and plasma cells. Immunohistochemical evaluation demonstrated that infiltrative cells in the early stage of myositis were CD8+ T-cells and that an increased expression of major histocompatibility complex (MHC) class I was apparent on the surface of nonnecrotic muscle fibers. In contrast, many CD3+ cells (T cells) and HLA-DR-positive macrophages and B lymphocytes were found in the severely affected areas. These results suggest that both expression of MHC class I and CD8+ T-cell infiltration may play an important role in initiation of myositis. These histopathologic findings resemble those reported in naturally occurring polymyositis in humans.     

Availability of NMR microscopic observation of mouse embryo disorder: examination in malformations induced by maternal administration of retinoic acid

Nuclear magnetic resonance (NMR) microscopy is a magnetic resonance imaging method with enhanced spatial resolution due to the use of a high static magnetic field and high magnetic field gradients. It is considered to be a useful tool for non-invasive and continuous investigation of tissue and organs at the histological level. In this study, we applied NMR microscopy to assessment of morphology in mouse embryos using a developmental disorder model induced by retinoic acid administration. Pregnant mice were given 50 mg/kg all-trans retinoic acid at 8.5 dpc. Embryos were collected at several time points after treatment and examined by NMR microscopy after fixation. Two-dimensional and three-dimensional spin echo sequences were used. Tissue contrast on two-dimensional images changed according to length of repetition time and echo time, and also to developmental stage of embryos. Two-dimensional and three-dimensional images nondestructively demonstrated defects in development of the skeleton and soft tissue, e.g. hypoplasia of vertebrae in the lumbar and tail regions and dysplasia of the spinal cord, in embryos exposed to retinoic acid. These morphological abnormalities were confirmed by conventional assessment after imaging. Although further improvements are required, NMR microscopy will provide a new approach for multi-parameter assessment of embryonic development under physiological and pathological conditions.     

Complex vertebral malformation in a stillborn Holstein calf in Japan

A female stillborn Holstein calf with shortened cervical and thoracic regions, protrusion of the tongue, and bilateral symmetric flexural contraction of the anterior limbs was delivered on gestation day 281. Multiple hemivertebrae, fused and misshaped vertebrae, synostosis and scoliosis of cervical, thoracic and lumbar vertebral column were found in the affected calf by radiographic and computed tomographic (CT) analysis. Ten pairs of ribs were present and the sternum consisted of 9 sternebrae. Multiple morphologic abnormalities including fusion, malformation, and displacement, were found in the ribs and sternum. Cardiac anomalies, including atrial septal defect and hypertrophy of right ventricle, were observed. DNA-polymerase chain reaction (PCR) analysis demonstrated that amplified product from the liver DNA of the affected calf had identical pattern to that associated with complex vertebral malformation (CVM) of Holstein calves and that her dam was a heterozygous carrier of CVM. The affected calf was diagnosed as having CVM based on the DNA-PCR results and the characteristic findings, and was recorded as a first documentation of CVM confirmed in a Holstein calf in Japan.     

Pythium rot of Chinese cabbage (Brassica rapa L. subsp. pekinensis) caused by Pythium aphanidermatum

Severe rot was found at the base of leaves and stems of Chinese cabbage (Brassica rapa L. subsp. pekinensis) in Ibaraki Prefecture every year in early September from 2002 through 2004. The causal fungus was identified as Pythium aphanidermatum (Edson) Fitzpatrick. This is the first report of P. aphanidermatum on Chinese cabbage. A similar disease of Chinese cabbage caused by P. ultimum Trow var. ultimum is known as Pythium rot. We propose adding P. aphanidermatum as a new pathogen of this disease.     

Primary malignant peripheral nerve sheath tumor with eosinophilic cytoplasmic globules arising from the greater omentum in a dog

A 10-year-old Golden Retriever dog had a solitary tumor mass arising from the greater omentum. Histologically, the tumor showed varying cellularity and patterns of cellular arrangement. In dense cellular areas, spindle-shaped cells were arranged in interlacing bundles. The sparse cellular area was characterized by loosely arranged fusiform cells. The neoplastic cells frequently contained PAS-positive eosinophilic globules in the cytoplasm, and mitotic figures were frequently observed. The tumor cells were positive to vimentin, S-100 protein, glial fibrillary acidic protein, myelin basic protein, neuron-specific enolase and myoglobin. The present tumor was diagnosed as a malignant peripheral nerve sheath tumor (MPNST) with eosinophilic cytoplasmic globules arising from the greater omentum. To our knowledge, this may be the first case of primary omental MPNST in domestic animals.     

Corrosive-wear characteristics of diamond-coated cemented carbide tools

Continuous milling tests with diamond-coated and uncoated cemented carbide tools and polycrystalline diamond tools were carried out on air-dried and wet melapi (Shorea sp.) and western redcedar (Thuja plicata D. Don). These tools were examined for corrosive-wear characteristics of the tool-edge appearance, cutting-edge profile, edge recession, and cutting-power consumption. The tool surfaces were observed with a scanning electron microscope and were analyzed by energy-dispersive X-ray spectroscopy. Based on these examinations, the occurrence of corrosive wear while cutting wet woods was confirmed for the uncoated and polycrystalline diamond tools. In contrast, the coated tools did not exhibit corrosive wear, nor did delamitation or wear of the diamond film occur with any of the work materials. The diamond-coated tools showed high resistance not only to mechanical wear but also to corrosive wear.The results of this work were presented at the 48th annual meeting of the Japan Wood Research Society, Shizuoka, April 1998     

Application of Pregerminated Brown Rice for Breadmaking

Pregerminated brown rice (PGBR) prepared by immersing in water was used for breadmaking, and effects on the dough properties and bread qualities were studied to compare with the ungerminated brown rice (BR). The substitution of BR or PGBR for wheat flour produced smaller specific volume in bread than in the control bread without BR or PGBR along with the increasing amount of substitution. However, the bread samples containing BR or PGBR suppressed staling during storage. The improving effect was especially obvious for substitutions of 10 and 20% PGBR as compared with BR. PGBR made viscous dough and retarded the staleness of bread compared with BR. γ‐Aminobutyric acid and oryzanol did not contain in the final BR and PGBR substituted bread, and phytic acid was decomposed ≈54 and 45% for 30% BR and 30% PGBR substituted breads, respectively. But ferulic acid was quite stable in the final baked product. As a result, substitution of PGBR for wheat flour improved the bread quality.     

Identification of enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri by expressed sequence tag analysis

Expressed sequence tag (EST) analyses were performed with the aim of identifying enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri. Of the 768 ESTs identified and sequenced, 669 were grouped into 324 clusters. Of these clusters, 123 comprising 245 ESTs were found to be homologous to genes reported to date. Among these, 43 clusters were annotated as enzymes according to the Enzyme Commission (EC) numbering system. Two EC groups, oxidoreductases and hydrolases, possessed a large number of ESTs. A cluster homologous to the glutathione peroxidase, an enzyme in the oxidoreductase group, contained 16 ESTs, which accounted for 2.4% of the total ESTs sequenced. There are three serine proteases, three cathepsins, two triacylgricerol lipases, and two chitinases among the clusters homologous to the enzymes in the hydrolase group. Since the squid liver functions in the digestive process, these enzymes would be involved in food digestion. Our data provide information on the various types of enzymes expressed in the squid liver and may provide a useful basis for further characterization of these enzymes.