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author:("Yanai, okuma")
1.  Pathological and Immunohistochemical Findings of Natural Highly Pathogenic Avian Influenza Infection in Tufted Ducks during 2010–2011 Outbreaks in Japan 
In the winter of 2010–2011, an outbreak of highly pathogenic avian influenza virus (HPAIV) infection occurred in wild and domestic birds in Japan. Tufted ducks were found dead in an urban area of Toyota City, Koriyama, Fukushima Prefecture. Two tufted ducks were examined histopathologically, immunohistochemically and molecularly. Gross findings included marked dark-red clotted blood in the pectoral muscles and multifocal hemorrhages on the serous membranes. Microscopically, non-suppurative meningoencephalitis, multifocal to coalescing pancreatic necrosis and severe pulmonary congestion were observed. HPAIV antigen was detected in the malacic areas, neuronal, glial and ependymal cells, pulmonary capillary endothelial cells and epithelium of pulmonary bronchioles, necrotic pancreatic acini and degenerated cardiac myocytes. The HPAIV isolate was genetically classified into clade group A. The broad distribution of virus antigen in brain and pulmonary tissues associated with HPAIV spontaneous infection in tufted ducks might be useful in understanding its pathogenesis in nature.
PMCID: PMC4197160  PMID: 24881650
HPAIV; Japan; outbreak; tufted duck
2.  Squamous Cell Carcinoma in a Capybara (Hydrochoerus hydrochaeris) 
A 4-year and 2-month-old male capybara (Hydrochoerus hydrochaeris) was diagnosed with squamous cell carcinoma on the buttocks after chronic recurrent dermatosis. The capybara was euthanized, examined by computed tomography and necropsied; the tumor was examined histologically. Computed tomography showed a dense soft tissue mass with indistinct borders at the buttocks. Histological examination of the tumor revealed islands of invasive squamous epithelial tumor cells with a severe desmoplastic reaction. Based on the pathological findings, the mass was diagnosed as a squamous cell carcinoma. This is the first study to report squamous cell carcinoma in a capybara.
PMCID: PMC4197163  PMID: 24909968
capybara; computed tomography; Hydrochoerus hydrochaeris; squamous cell carcinoma
3.  Infectivity of Cryptosporidium andersoni and Cryptosporidium muris to Normal and Immunosuppressive Cynomolgus Monkeys 
Cryptosporidium andersoni and Cryptosporidium muris infections have been found in the mice and/or cattle. The oocysts of C. andersoni and C. muris have been sporadically detected in human feces, but the infectious capacity and features have been unknown, because of the scarcity of reports involving human infections. To assess the infectivity and the clinical and pathological features of C. andersoni and C. muris in primates, an experimental infectious study was conducted using cynomolgus monkeys. The monkeys were orally inoculated with oocysts of two different C. andersoni Kawatabi types and C. muris RN-66 under normal and immunosuppressive conditions. The feces of the monkeys were monitored for about 40 days after the administration of oocysts using the flotation method, but no shedding oocysts were observed under either both normal or immunosuppressive conditions. Gross and histopathological examinations were performed on the immunosuppressive monkeys, but these revealed no evidence of Cryptosporidium infections, even though the monkeys were subjected to immunosuppressive conditions. It is hypothesized that C. andersoni and C. muris pose little danger of infection in primates even under immunosuppressive conditions.
PMCID: PMC3982829  PMID: 24132296
Cryptosporidium; experimental animals; monkey; parasitology; pathology
4.  Hypodectes propus (Acarina: Hypoderatidae) in a rufous turtle dove, Streptopelia orientalis (Aves: Columbiformes), in Japan 
Journal of Veterinary Science  2013;14(4):421-424.
An adult male rufous turtle dove, Streptopelia (S.) orientalis (Aves: Columbiformes), was found dead in Yorii-machi Town, Osato District 369-1217, Saitama Prefecture, Japan, and subjected to necropsy. A large number of immobile hypopi (deutonymphs) of the hypoderatid mite, Hypodectes (H.) propus (Acarina: Hypoderatidae), were found individually encapsulated subcutaneously primarily in the adipose tissue. The mites were 1.43 mm in length and 0.44 mm in width on average, and had provoked mild inflammatory reactions that predominantly manifested as foamy macrophages and lymphoplasmocytes. PCR analysis using ribosomal DNA extracted from paraffin-blocked tissues produced a 240 bp band specific for hypoderatids. Based on the morphological features (distinct coxal apodemes, especially in the anterior portion) and PCR-based findings, the hypopi were identified as H. propus. To the best of our knowledge, this is the first case describing the subcutaneous mite H. propus in a rufous turtle dove, S. orientalis, in Japan. This study also highlights the use of paraffin blocks as a source of tissue DNA for molecular evaluation.
PMCID: PMC3885735  PMID: 23820220
Hypodectes propus; Japan; Strepopelia orientalis; subcutaneous tissue
5.  Coenurus cerebralis Cysts in the Left Lateral Cerebral Ventricle of a Ewe 
A three-and-a-half year-old female Rahmani ewe was presented suffering from nervous symptoms. Grossly, a large cyst measuring 7 × 4 cm and weighing 145 g occupied the dilated left lateral ventricle. The overlying cerebral tissue was thin, atrophied and congested. It tore easily, and the cyst was evacuated spontaneously. Microscopically, liquefactive necrosis surrounded by aggregations of macrophages, eosinophils, lymphocytes, fibroblasts and giant cells was predominantly observed. Hyperplasia and severe necrosis of the ependymal cell lining of the lateral ventricle were observed. Extensive subependymal inflammatory cell infiltrations, accompanied by neovascularization and fibroblastic proliferation, were seen. Based on the gross and histopathological lesions and cyst morphology and location, the cyst was diagnosed as Coenurus cerebralis. This report describes a rare case of coenurus cyst in the left lateral cerebral ventricle of a ewe and the associated lesion.
PMCID: PMC3942954  PMID: 23884082
cerebral; Coenurus; cyst; ewe; ventricle
6.  Constitutive phosphorylation of the mTORC2/Akt/4E-BP1 pathway in newly derived canine hemangiosarcoma cell lines 
Canine hemangiosarcoma (HSA) is a malignant tumor with poor long-term prognosis due to development of metastasis despite aggressive treatment. The phosphatidyl-inositol-3 kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway is involved in its endothelial pathologies; however, it remains unknown how this pathway plays a role in canine HSA. Here, we characterized new canine HSA cell lines derived from nude mice-xenografted canine HSAs and investigated the deregulation of the signaling pathways in these cell lines.
Seven canine HSA cell lines were established from 3 xenograft canine HSAs and showed characteristics of endothelial cells (ECs), that is, uptake of acetylated low-density lipoprotein and expression of canine-specific CD31 mRNA. They showed varied morphologies and mRNA expression levels for VEGF-A, bFGF, HGF, IGF-I, EGF, PDGF-B, and their receptors. Cell proliferation was stimulated by these growth factors and fetal bovine serum (FBS) in 1 cell line and by FBS alone in 3 cell lines. However, cell proliferation was not stimulated by growth factors and FBS in the remaining 3 cell lines. Phosphorylated p44/42 Erk1/2 was increased by FBS stimulation in 4 cell lines. In contrast, phosphorylation of Akt at Ser473, mTOR complex 1 (mTORC1) at Ser2448, and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) at Ser65 was high in serum-starved condition and not altered by FBS stimulation in 6 cell lines, despite increased phosphorylation of these residues in normal canine ECs. This suggested that the mTORC2/Akt/4E-BP1 pathway was constitutively activated in these 6 canine HSA cell lines. After cell inoculation into nude mice, canine HSA tumors were formed from 4 cell lines and showed Akt and 4E-BP1 phosphorylation identical to the parental cell lines.
Our findings suggest that the present cell lines may be useful tools for investigating the role of the mTORC2/Akt/4E-BP1 pathway in canine HSA formation both in vivo and in vitro.
PMCID: PMC3438112  PMID: 22839755
Cell line; Canine; Akt; Hemangiosarcoma; mTOR; 4E-BP1
7.  Characterization of Squamate Olfactory Receptor Genes and Their Transcripts by the High-Throughput Sequencing Approach 
Genome Biology and Evolution  2012;4(4):602-616.
The olfactory receptor (OR) genes represent the largest multigene family in the genome of terrestrial vertebrates. Here, the high-throughput next-generation sequencing (NGS) approach was applied to characterization of OR gene repertoires in the green anole lizard Anolis carolinensis and the Japanese four-lined ratsnake Elaphe quadrivirgata. Tagged polymerase chain reaction (PCR) products amplified from either genomic DNA or cDNA of the two species were used for parallel pyrosequencing, assembling, and screening for errors in PCR and pyrosequencing. Starting from the lizard genomic DNA, we accurately identified 56 of 136 OR genes that were identified from its draft genome sequence. These recovered genes were broadly distributed in the phylogenetic tree of vertebrate OR genes without severe biases toward particular OR families. Ninety-six OR genes were identified from the ratsnake genomic DNA, implying that the snake has more OR gene loci than the anole lizard in response to an increased need for the acuity of olfaction. This view is supported by the estimated number of OR genes in the Burmese python's draft genome (∼280), although squamates may generally have fewer OR genes than terrestrial mammals and amphibians. The OR gene repertoire of the python seems unique in that many class I OR genes are retained. The NGS approach also allowed us to identify candidates of highly expressed and silent OR gene copies in the lizard's olfactory epithelium. The approach will facilitate efficient and parallel characterization of considerable unbiased proportions of multigene family members and their transcripts from nonmodel organisms.
PMCID: PMC3342882  PMID: 22511035
next-generation sequencing; olfactory receptor; Squamata; molecular evolution; pseudogene
8.  Establishment of canine hemangiosarcoma xenograft models expressing endothelial growth factors, their receptors, and angiogenesis-associated homeobox genes 
BMC Cancer  2009;9:363.
Human hemangiosarcoma (HSA) tends to have a poor prognosis; its tumorigenesis has not been elucidated, as there is a dearth of HSA clinical specimens and no experimental model for HSA. However, the incidence of spontaneous HSA is relatively high in canines; therefore, canine HSA has been useful in the study of human HSA. Recently, the production of angiogenic growth factors and their receptors in human and canine HSA has been reported. Moreover, the growth-factor environment of HSA is very similar to that of pathophysiological angiogenesis, which some homeobox genes regulate in the transcription of angiogenic molecules. In the present study, we established 6 xenograft canine HSA tumors and detected the expression of growth factors, their receptors, and angiogenic homeobox genes.
Six primary canine HSAs were xenografted to nude mice subcutaneously and serially transplanted. Subsequently, the expressions of vascular endothelial growth factor (VEGF)-A, basic fibroblast growth factors (bFGF), flt-1 and flk-1 (receptors of VEGF-A), FGFR-1, and angiogenic homeobox genes HoxA9, HoxB3, HoxB7, HoxD3, Pbx1, and Meis1 were investigated in original and xenograft tumors by histopathology, immunostaining, and reverse transcription polymerase chain reaction (RT-PCR), using canine-specific primer sets.
Histopathologically, xenograft tumors comprised a proliferation of neoplastic cells that were varied in shape, from spindle-shaped and polygonal to ovoid; some vascular-like structures and vascular clefts of channels were observed, similar to those in the original tumors. The expression of endothelial markers (CD31 and vWF) was detected in xenograft tumors by immunohistochemistry and RT-PCR. Moreover, the expression of VEGF-A, bFGF, flt-1, flk-1, FGFR-1, HoxA9, HoxB3, HoxB7, HoxD3, Pbx1, and Meis1 was detected in xenograft tumors. Interestingly, expressions of bFGF tended to be higher in 3 of the xenograft HSA tumors than in the other tumors.
We established 6 xenograft canine HSA tumors in nude mice and found that the expressions of angiogenic growth factors and their receptors in xenograft HSAs were similar to those in spontaneous HSA. Furthermore, we detected the expression of angiogenic homeobox genes; therefore, xenograft models may be useful in analyzing malignant growth in HSA.
PMCID: PMC2768746  PMID: 19825192
9.  Proliferative Potential of a Spinal Nephroblastoma in a Young Dog 
The proliferative potential of a spinal nephroblastoma was studied in a young dog. A 4-month-old, female golden retriever showed developing deterioration in her gait and subsequent paralysis of her hind legs. At necropsy, a well-demarcated grayish brown tumor mass was found in the lumbar spinal cord segments between L2 and L3. Histologically, a blastemal cell tumor with a tubule- or glomeruli-like structure was found to be infiltrating intradurally. Proliferating cells at the S-phase, assessed using the bromodeoxyuridine (BrdU) labeling method, were seen occasionally in the tubular cells and glomeruli-like structures and were frequently seen in the blastemal cells. Immunohistochemically, the tubular epithelial cells were positive for cytokeratin, and the blastemal cells were positive for vimentin. The present tumor showed a high potential for growth and invasion, which suggests that it the potential to expand into the adjacent spinal cord.
PMCID: PMC3246021  PMID: 22271979
nephroblastoma; spinal cord; young dog; BrdU labeling; S-phase cells; immunohistochemistry
10.  Nocardia africana Isolated from a Feline Mycetoma 
Journal of Clinical Microbiology  2003;41(2):908-910.
Nocardia africana was isolated from a subcutaneous nodule of a cat. The isolate developed orange, wrinkled colonies. The bacteria were rod shaped to coccoid (1 by 5 μm) and gram positive. Analysis of the 16S ribosomal DNAs of the isolate and a reference strain of N. africana showed 100% homology.    
PMCID: PMC149653  PMID: 12574314
11.  Characteristics of a Novel Type of Bovine Cryptosporidium andersoni 
We isolated oocysts that resemble Cryptosporidium andersoni from cattle grazing on a farm in Japan. The partial sequences of genes from the isolate were coincident with published sequences of genes of C. andersoni. Since the isolate was able to infect SCID mice, the isolate appears to be a novel type of C. andersoni.
PMCID: PMC152376  PMID: 12514063

Results 1-11 (11)