〔主要な業績〕糞の中の細菌は、腸の定着菌か、外部からの通過菌なのか －糞から精製したDNAで分かることは何か－　　　　　　　　　　　　　　　
上塚浩司, 筆頭著者
動物園学紀要, 2024年03月

採卵鶏での実験的誘導換羽における鰓後腺の形態学的変化の観察　　　　　　　　　　　　　　　
北島克好; 上塚浩司; 中島弘美
日本家禽学会誌, 2023年10月, ［査読有り］

採卵鶏の産卵期における鰓後腺の形態変化の経時的観察　　　　　　　　　　　　　　　
北島克好; 上塚浩司; 中島弘美
日本家禽学会誌, 2023年04月, ［査読有り］

〔主要な業績〕茨城県の野生の留鳥におけるClostridium perfringensの保有状況　　　　　　　　　　　　　　　
関光一朗、鈴木菜月、永井あゆみ、仲本あゆ美、山本有花、鎌塚倫成、木村和輝、菊地大希、上塚浩司, ラスト(シニア)オーサー
家畜衛生学会誌, 2022年01月, ［査読有り］

サラブレッド種競走馬の感染性皮膚炎からの細菌・真菌の分離同定と薬剤感受性調査　　　　　　　　　　　　　　　
牛屋 重人、上塚 浩司, 日本家畜衛生学会
家畜衛生学雑誌, 2021年07月29日, ［査読有り］

Intracellular localization of Mycoplasma bovis in the bronchiolar epithelium of experimentally infected calves.
Nunoya T; Omori T; Tomioka H; Umeda F; Suzuki T; Uetsuka K, ラスト(シニア)オーサー
Journal of Comparative Pathology, 2020年03月, ［査読有り］

ITS-1領域を基にしたPCR法によるカラス腸内容物からのEimeria属原虫の検出とその分子系統解析　　　　　　　　　　　　　　　
菊地大希、成廣隆、木村和輝、上塚浩司, ラスト(シニア)オーサー, 日本家畜衛生学会
家畜衛生学雑誌, 2019年12月, ［査読有り］

Proliferative changes in the adrenal medulla of aged Chinese native pigs.
Koji Uetsuka; Takayuki Suzuki; James Kenn Chambers; Kazuyuki Uchida; Hiroyuki Nakayama; Kunio Doi; Tetsuo Nunoya, Four aged retired Chinese native pigs, three females and one male, estimated as over 10-year-old, were subjected to autopsy because of infertility due to aging. Grossly, nodular lesions were found bilaterally in the adrenal medulla of all four pigs. Based on the gross and the histopathological findings, they were diagnosed as either medullary nodular hyperplasia or pheochromocytoma. Immunohistochemically, proliferating cells of all these lesions were immuno-positive for chromogranin-A, indicating adrenal medulla-derived. Ultrastructurally, cytoplasmic neurosecretory granules suggestive of secretion were observed in these proliferating cells. There have been only limited numbers of reports on adrenal medullar proliferative changes including pheochromocytoma in pigs. The present cases will provide a valuable information for the characterization of similar changes in animals and human., 公益社団法人 日本獣医学会
The Journal of veterinary medical science, 2018年06月29日, ［査読有り］

Experimental Induction of necrotic enteritis in chickens by a netB-positive Japanese isolate of Clostridium perfringens
Ho To; Takayuki Suzuki; Fumiya Kawahara; Koji Uetsuka; Shinya Nagai; Tetsuo Nunoya, Necrotic enteritis (NE) is one of the most important bacterial diseases in terms of economic losses. Clostridium perfringens necrotic enteritis toxin B, NetB, was recently proposed as a new key virulent factor for the development of NE. The goal of this work was to develop a necrotic enteritis model in chickens by using a Japanese isolate of C. perfringens. The Japanese isolate has been found to contain netB gene, which had the same nucleotide and deduced amino acid sequences as those of prototype gene characterized in Australian strain EHE-NE18, and also expressed in vitro a 33-kDa protein identified as NetB toxin by nano-scale liquid chromatographic tandem mass spectrometry. In the challenge experiment, broiler chickens fed a commercial chicken starter diet for 14 days post-hatch were changed to a high protein feed mixed 50:50 with fishmeal for 6 days. At day 21 of age, feed was withheld for 24 hr, and each chicken was orally challenged twice daily with 2 m/each of C. perfringens culture (10(9) to 10(10) CFU) on 5 consecutive days.The gross necrotic lesions were observed in 90 and 12.5% of challenged and control chickens, respectively. To our knowledge, this is the first study that demonstrated that a netB-positive Japanese isolate of C. perfringens is able to induce the clinical signs and lesions characteristic of NE in the experimental model, which may be useful for evaluating the pathogenicity of field isolates, the efficacy of a vaccine or a specific drug against NE., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2017年02月, ［査読有り］

Identification of a Unique Amyloid Sequence in AA Amyloidosis of a Pig Associated With Streptococcus Suis Infection
J. Kamiie; G. Sugahara; S. Yoshimoto; N. Aihara; T. Mineshige; K. Uetsuka; K. Shirota, Here we report a pig with amyloid A (AA) amyloidosis associated with Streptococcus suis infection and identification of a unique amyloid sequence in the amyloid deposits in the tissue. Tissues from the 180-day-old underdeveloped pig contained foci of necrosis and suppurative inflammation associated with S. suis infection. Congo red stain, immunohistochemistry, and electron microscopy revealed intense AA deposition in the spleen and renal glomeruli. Mass spectrometric analysis of amyloid material extracted from the spleen showed serum AA 2 (SAA2) peptide as well as a unique peptide sequence previously reported in a pig with AA amyloidosis. The common detection of the unique amyloid sequence in the current and past cases of AA amyloidosis in pigs suggests that this amyloid sequence might play a key role in the development of porcine AA amyloidosis. An in vitro fibrillation assay demonstrated that the unique AA peptide formed typically rigid, long amyloid fibrils (10 nm wide) and the N-terminus peptide of SAA2 formed zigzagged, short fibers (7 nm wide). Moreover, the SAA2 peptide formed long, rigid amyloid fibrils in the presence of sonicated amyloid fibrils formed by the unique AA peptide. These findings indicate that the N-terminus of SAA2 as well as the AA peptide mediate the development of AA amyloidosis in pigs via cross-seeding polymerization., SAGE PUBLICATIONS INC
VETERINARY PATHOLOGY, 2017年01月, ［査読有り］

Tumor-Free Transplantation of Patient-Derived Induced Pluripotent Stem Cell Progeny for Customized Islet Regeneration
Moustafa M. El Khatib; Seiga Ohmine; Egon J. Jacobus; Jason M. Tonne; Salma G. Morsy; Sara J. Holditch; Claire A. Schreiber; Koji Uetsuka; Noemi Fusaki; Dennis A. Wigle; Andre Terzic; Yogish C. Kudva; Yasuhiro Ikeda, Human induced pluripotent stem cells (iPSCs) and derived progeny provide invaluable regenerative platforms, yet their clinical translation has been compromised by their biosafety concern. Here, we assessed the safety of transplanting patient-derived iPSC-generated pancreatic endoderm/progenitor cells. Transplantation of progenitors from iPSCs reprogrammed by lentiviral vectors (LV-iPSCs) led to the formation of invasive teratocarcinoma-like tumors in more than 90% of immunodeficient mice. Moreover, removal of primary tumors from LV-iPSC progeny-transplanted hosts generated secondary and metastatic tumors. Combined transgene-free (TGF) reprogramming and elimination of residual pluripotent cells by enzymatic dissociation ensured tumor-free transplantation, ultimately enabling regeneration of type 1 diabetes-specific human islet structures in vivo. The incidence of tumor formation in TGF-iPSCs was titratable, depending on the oncogenic load, with reintegration of the cMYC expressing vector abolishing tumor-free transplantation. Thus, transgene-free cMYC-independent reprogramming and elimination of residual pluripotent cells are mandatory steps in achieving transplantation of iPSC progeny for customized and safe islet regeneration in vivo., WILEY
STEM CELLS TRANSLATIONAL MEDICINE, 2016年05月

Drug-susceptibility of isolates of Brachyspira hyodysenteriae isolated from colonic mucosal specimens of pigs collected from slaughter houses in Japan in 2009
Keita Kajiwara; Midori Kozawa; Takuya Kanazawa; Kouji Uetsuka; Hiromi Nakajima; Yoshikazu Adachi, Twenty nine isolates identified as Brachyspira hyodysenteriae were most susceptible to carbadox and metronidazole, whereas they were resistant to macrolides. The isolates showed intermediate susceptibility to tiamulin, lincomycin, penicillin G, ampicillin, chloramphenicol, tetracycline, enrofloxacin and valnemulin, with MIC50 values ranging from 0.39 to 3.13., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2016年03月, ［査読有り］

Malignant Sertoli Cell Tumor in a Goose (Anser cygnoides domesticus)
Koji Uetsuka; Takayuki Suzuki; Kunio Doi; Tetsuo Nunoya, This paper describes the pathologic features of a malignant Sertoli cell tumor found in an adult goose (Anser cygnoides domesticus). At necropsy, in addition to one large tumor mass (15 cm in diameter), multiple small tumor masses were observed over the peritoneum and mesenterium in the coelomic cavity. The large tumor mass was composed of sheets, lobules, and small islands of tumor cells, and elongated tumor cells lying perpendicular to fibrous connective tissue were characteristic. Such histopathologic characteristics were common to all the tumors. The tumor cells were immunohistochemically positive for neuron-specific enolase and S-100, and some tumor cells contained fine intracytoplasmic pigments that stained red by oil red O staining. These findings, taken together with the fact that one testis was markedly atrophied and bore no tumor cells and the other testis was not discernible, the present case was diagnosed as unilateral malignant Sertoli cell tumor arising from the unilateral testis. To our knowledge, this is the first report of Sertoli cell tumor in the goose., AMER ASSOC AVIAN PATHOLOGISTS
AVIAN DISEASES, 2012年12月, ［査読有り］

高齢の台湾小耳種豚でみられた副腎髄質の増殖性病変の病理学的解析　　　　　　　　　　　　　　　
上塚 浩司; 鈴木 敬之; 富岡 ひとみ; チェンバーズ・ジェームズ; 内田 和幸; 中山 裕之; 土井 邦雄; 布谷 鉄夫, (公社)日本獣医学会
日本獣医学会学術集会講演要旨集, 2012年08月, ［査読有り］

A Case of Renal Oxalosis in a 3-Month-Old Cat Raised under Controlled Conditions
Takayuki Suzuki; Koji Uetsuka; Kunio Doi; Tetsuo Nunoya, The kidneys of a 3-month-old female cat were examined. The cat which had been raised under controlled conditions with no history of any poisoning showed progressive weight loss with increases in blood BUN and creatinine concentrations. At necropsy, both kidneys were firm in consistency with formation of focal scars. Histopathologically, widespread deposition of crystals was observed in the renal tubules (in both dilated lumina and degenerative epithelia) accompanying mild interstitial fibrosis with lymphocyte infiltration. The crystals were colorless or basophilic on the hematoxilin and eosin-stained section and could be visualized with polarized light as doubly fractile crystals. The crystals were identified as calcium oxalate crystals by histochemical examinations using von Kossa stain and alizarin red S stain under different conditions and by ultrastructural examination. Judging from the above-mentioned findings, the present renal lesion detected in an infant cat was diagnosed as renal oxalosis which was suspected to be hereditary in nature., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2012年03月, ［査読有り］

A Case of Congenital Hyperostosis in a Newborn Piglet
Koji Uetsuka; Takayuki Suzuki; Hirohito Ogawa; Hiroko Sato; Kunio Doi; Tetsuo Nunoya, One female newborn piglet showed prominent thickening of both forelimbs and died soon after birth. Histopathologically, thin and woven trabeculae of bone was extending out from the edge of cortical bone in the affected forelimbs, and diagnosed as congenital hyperostosis. The extent of radially proliferated trabeculae was most prominent in radioulna. Many round- to spindle-shaped cells were observed in periosteum, which were considered to be osteoblasts. Around the periosteum, the mesenchymal proliferation was extensive with abundant mucus, and cartilaginous metaplastic changes were observed mainly around the radioulna and humerus. Dilatation of vessels with fibrin deposition in vessel walls was often observed, which were considered to reflect the localized circulatory disturbance., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2012年02月, ［査読有り］

Clinical and Pathologic Features of Neuronal Ceroid-Lipofuscinosis in a Ferret (Mustela putorius furo)
K. Nibe; Y. Miwa; S. Matsunaga; J. K. Chambers; K. Uetsuka; H. Nakayama; K. Uchida, Clinical and pathologic features of neuronal ceroid-lipofuscinosis in a 4-month-old ferret are reported. Clinical signs including neurological symptoms appeared at 3 months of age and progressed rapidly. By magnetic resonance imaging, severe cerebral atrophy was recognized. Histopathologically, there was severe neuronal loss and diffuse astrogliosis with macrophage accumulations; lesions were found predominantly in the cerebral cortex. Intracytoplasmic pigments were observed in surviving neurons and macrophages throughout the brain. The pigments were intensely positive for periodic acid-Schiff, Luxol fast blue, and Sudan black B and exhibited a green autofluorescence. Electron microscopic examination revealed the accumulation of electron-dense granular material within lysosomes of neurons and macrophages. Immunohistochemically, a large number of saposin-positive granules accumulated in the neuronal cells, astrocytes, and macrophages of the lesions, but significant immunoreactivity for subunit c of mitochondrial adenosine triphosphate synthase was not observed. Based on these findings, the animal was diagnosed as affected by neuronal ceroid-lipofuscinosis., SAGE PUBLICATIONS INC
VETERINARY PATHOLOGY, 2011年11月, ［査読有り］

Proventricular Dilatation Disease Associated with Avian Bornavirus Infection in a Citron-Crested Cockatoo that Was Born and Hand-Reared in Japan
Hirohito Ogawa; Yasuyuki Sanada; Naoko Sanada; Megumi Kudo; Kotaro Tuchiya; Toshiaki Kodama; Koji Uetsuka, A 5-month-old female Citron-crested Cockatoo (Cacatua sulphurea citrinocristata) that was born and hand-reared in Japan died with suspected proventricular dilatation disease (PDD). Macroscopic and microscopic examinations of the bird revealed characteristic features of PDD, i.e., distention of the proventriculus and infiltration of lymphocytes and plasma cells in ganglia of various organs and in central and peripheral nerves. A linkage of this PDD case to infection with avian bornavirus (ABV) was documented by RTPCR amplification of the virus genomes from the affected bird. Phylogenetic analysis revealed that the ABV identified in this study clustered into the genotype 2, which is one of the dominant ABV genotypes worldwide. To the best of our knowledge, this is the first report of a natural case of PDD associated with ABV infection in Japan., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2011年06月, ［査読有り］

Hepatic Myelolipoma with Systemic Amyloidosis in a Goose (Anser cygnoides domesticus)
Takayuki Suzuki; Kouji Uetsuka; Kouichi Kusanagi; Takuya Hirai; Tetsuo Nunoya; Kunio Doi, We report here a case of hepatic myelolipoma with systemic amyloidosis in a goose (Anser cygnoides domesticity), which died suddenly following the short history of weakness and greenish diarrhea. At necropsy, multiple yellowish-white foci were observed on the surface of the prominently enlarged liver. Histologically, there were multiple foci of adipose tissue admixed with myeloid elements in various proportions in the liver as well as amyloid deposition in several organs including the liver, intestine, spleen, kidney, and ovary. Ultrastructurally, erythroblast-like cells and myelocytes, which showed various stages of differentiation, were observed in the foci of the liver. These findings shared characteristics of hepatic myelolipoma which is very rare in birds., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2010年05月, ［査読有り］

Disseminated Histiocytic Sarcoma with Excessive Hemophagocytosis in a Cat
Kaori Ide; Asuka Setoguchi-Mukai; Takayuki Nakagawa; Koji Uetsuka; Hiroyuki Nakayama; Yasuhito Fujino; Koichi Ohno; Hajime Tsujimoto, A 10-year-old Japanese domestic cat was presented with anorexia and weight loss. Severe anemia and thrombocytopenia were detected. Abdominal radiography and Ultrasonography revealed the presence of multiple masses in the spleen. Cytological analyses of the masses revealed several atypical histiocytic cells and considerable hemophagocytosis. A splenectomy was performed, and the mass was diagnosed as histiocytic Sarcoma on the basis of histopathological, cytochemical and immunohistochemical analyses. Further, abnormal hemophagocytosis was observed in the bone marrow. The cat was administered prednisolone and lomustine, and it survived for 107 days after admission. An autopsy revealed the presence of neoplastic histiocytic cells in the bone marrow, liver, pancreatic lymph node and glomeruli. This is the first case of histiocytic Sarcoma in a cat to be reported in Japan., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2009年06月, ［査読有り］

Etoposide Induces TRP53-Dependent Apoptosis and TRP53-Independent Cell Cycle Arrest in Trophoblasts of the Developing Mouse Placenta
Hirofumi Yamauchi; Kei-ichi Katayama; Masaki Ueno; Hiroyuki Kanemitsu; Chunja Nam; Takashi Mikami; Aya Saito; Yuka Ishida; Koji Uetsuka; Kunio Doi; Yasushi Ohmach; Hiroyuki Nakayama, Abnormal regulation of placental apoptosis and proliferation has been implicated in placental disorders. Recently, several DNA-damaging agents were reported to induce excessive apoptosis and reduce cell proliferation in the placenta; however, the molecular pathways of these toxic effects on the placenta are unclear. The aim of the present study was to determine the involvement of TRP53, a tumor suppressor that mediates cellular responses to DNA damage, in the induction of apoptosis and cell cycle arrest in the developing placenta. For this purpose, we treated pregnant mice on Day 12 of gestation with 10 mg/kg of etoposide and 5-Gy gamma irradiation, potent inducers of DNA damage. We found an increase in the number of trophoblastic apoptoses 8 and 24 h after etoposide injection and 6 and 24 h after irradiation in the placental labyrinth zone. The number of mitoses and DNA syntheses in trophoblasts decreased after treatment. The accumulation and phosphorylation of TRP53 protein were detected 8 and 6 h after etoposide injection and irradiation, respectively. In Trp53-deficient placentas, the induction of etoposide-induced trophoblastic apoptosis is abrogated, while the reduction of proliferation occurred similarly as in wild-type placentas. CDC2A, a regulator of G2/M progression, was inactivated by phosphorylation after etoposide injection and irradiation, suggesting that the cell cycle was arrested at the G2/M border by treatment. Our study demonstrated that etoposide injection induced TRP53-dependent apoptosis and TRP53-independent cell cycle arrest in labyrinthine trophoblasts, providing insights into the molecular pathway of placental disorders., SOC STUDY REPRODUCTION
BIOLOGY OF REPRODUCTION, 2009年04月, ［査読有り］

Spontaneous cutaneous mast cell tumor with lymph node metastasis in a Richardson’s gournd squirrel (Spermophilus richardsonii).
He XJ; Uchida K; Tochitani T; Uetsuka K; Miwa Y; Nakayama H, A 4-year-old female Richardson's ground squirrel (Spermophilus richardsonii) presented with multicentric nodules arising from the skin of the middle of the tail and lumbosacral regions. Histologically, the nodules were composed of a proliferation of spindloid to pleomorphic cells that sometimes formed sheets and fascicular to storiform patterns. Diffuse infiltration of eosinophils was also noted. The results of immunohistochemistry indicated positive labeling for vimentin, mast cell tryptase, c-kit, and Ki-67. Toluidine blue stain revealed fine, metachromatic, cytoplasmic granules. The histologic diagnosis was mast cell tumor. The neoplasm recurred and metastasized to the right lumbar lymph node 1 month later., AMER ASSOC VETERINARY LABORATORY DIAGNOSTICIANS INC
Journal of Veterinary Diagnostic Investigation., 2009年01月, ［査読有り］

Neural progenitor cells are protected against MPTP by MAO-B inhibitors
Xi Jun He; Koji Uetsuka; Hiroyuki Nakayama, Neurotoxic effects of MPTP on the nigrostriatal dopaminergic system are thought to be initiated by 1-methyl-4-phenylpyridinium (MPP+), a metabolite formed by the monoamine oxidase (MAO)-B-mediated oxidation of MPTP. We previously reported that the administration of MPTP induced apoptosis in migrating neuroblasts (neural progenitor cells, NPCs) in adult mice. To determine whether MAO-B is also involved in the neurotoxicity of MPTP to NPCs, this study looked at the effects of MAO B inhibitors, R(-)-deprenyl (deprenyl) and N-(2-aminoethyl)-4-chlorobenzamide (Ro 16-6491), both of which protect the dopaminergic system against MPTP. Few apoptotic cells were found in saline- or MAO-B inhibitor-treated animals but MPTP markedly induced apoptosis in the subventricular zone (SVZ) and rostral migratory stream (RMS) after 1 day. When mice were pretreated with deprenyl or Ro 16-6491, not only nigrostriatal dopamine levels but also NPCs were significantly protected against MPTP. In addition, MPTP-induced apoptosis was found in both juvenile (postnatal 21 days) and older (12 months old) mice, suggesting NPCs to be different from the dopamine system, which has been thought to exhibit age-dependent susceptibility to MPTP. (C) 2008 Elsevier Inc. All rights reserved., ELSEVIER SCIENCE BV
NEUROTOXICOLOGY, 2008年11月, ［査読有り］

The Effect of Fasting on Hepatic Lipid Accumulation and Transcriptional Regulation of Lipid Metabolism Differs between C57BL/6J and BALB/cA Mice Fed a High-fat Diet
Satomi Nishikawa; Kunio Doi; Hiroyuki Nakayama; Koji Uetsuka, The effects of fasting on hepatic lipid metabolism in mice fed a high-fat diet (HFD) are still unclear. After fasting, the degree of hepatic lipid accumulation differs between HFD-fed C57BL/6J (B6) and BALB/cA (BALB/c) mice. It is not clear whether this difference is due to sensitivity to fasting or HFD. The aim of this study is to elucidate this difference among strains. After nine weeks of HFD feeding, both B6 and BALB/c mice showed moderate hepatic steatosis. However, after a subsequent twenty-hour fast, the hepatic lipid accumulation was markedly decreased in B6 but not in BALB/c mice. Moreover, the mRNA expression of a transcription factor, Srebp1 (regulates hepatic lipid metabolism), and its target genes-malic enzyme, acetyl-CoA carboxylase, fatty acid synthase (regulate fatty acid synthesis), and glycerol-3-phosphate acyltransferase (regulates triacylglycerol synthesis)-were more markedly reduced in B6 than BALB/c mice. In conclusion, fasting may modify hepatic lipid accumulation in HFD-fed B6 and BALB/c mice differently. The difference may be partly owing to a marked downregulation of the expression of some lipid-metabolism-related genes in B6 mice. These results suggest that fasting per se has a significant effect on hepatic lipid accumulation in mouse strains. SREBP1 might play a role in this fasting effect., SAGE PUBLICATIONS INC
TOXICOLOGIC PATHOLOGY, 2008年10月, ［査読有り］

Hepatic Lesions of A Standard Poodle Dog with Intrahepatic Portosystemic Shunt
Kaori Isobe; Satoru Matsunaga; Hiroyuki Nakayama; Koji Uetsuka, A 1-year-and-3-month-old, male standard poodle dog with intrahepatic portosystemic shunt (PSS) was autopsied. Nineteen regions of the liver were prepared for detailed examination, and the distribution of hepatic lesions caused by PSS was studied in the liver of this dog. Histopathologically, the liver revealed a variety of hepatic lesions including lipogranulomas in the hepatic parenchyma, and a ductular reaction and microvascular proliferation in portal areas. The distribution of the lesions was not significantly different among liver regions. It is concluded that, in the present case, hepatic lesions caused by PSS are independent of shunt location, and are distributed equally in the liver., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2008年10月

Hypereosinophilic Syndrome in Two Cats
Yoshinori Takeuchi; Shinobu Matsuura; Yasuhito Fujino; Mayumi Nakajima; Masashi Takahashi; Ko Nakashima; Yusuke Sakai; Koji Uetsuka; Koichi Ohno; Hiroyuki Nakayama; Hajime Tsujimoto, Two cats showing chronic vomiting, diarrhea and weight loss were found to have leukocytosis with marked eosinophilia. Both cats were diagnosed with hypereosinophilic syndrome by the findings of increased eosinophils and their precursors in the bone marrow, eosinophilic infiltration into multiple organs, and exclusion of other causes for eosinophilia. Although cytoreductive chemotherapy with hydroxycarbamide and prednisolone was performed, these two cats died 48 days and 91 days after the initial presentation., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2008年10月, ［査読有り］

Incorporation of beta-amyloid protein through the bovine ileal epithelium before and after weaning: Model for orally transmitted amyloidoses
Yasuhisa Ano; Hiroyuki Nakayama; Yusuke Sakai; Akikazu Sakudo; Maiko Endo; Shogo Ebisu; JunYou Li; Koji Uetsuka; Noboru Manabe; Takashi Onodera, To determine the mechanism of bovine intestinal incorporation of the pathogen, and the pathogenesis of prion protein in the early stage, cows suckling and weaning were orally given a fusion protein of A beta-EGFP. A beta-EGFP was incorporated through the villous columnar epithelial cells and accumulated in crypt patches in the ileum of suckling cows. The sites of the uptake and accumulation of A beta-EGFP are very close to the peripheral nervous system; however, such uptake of A beta-EGFP was not observed in 6-month-old post-weaning cows. The present study, therefore, suggests that the weaning period is very important for the risk of transmission., BLACKWELL PUBLISHING
MICROBIOLOGY AND IMMUNOLOGY, 2008年08月, ［査読有り］

Complementary distributions of amyloid-beta and neprilysin in the brains of dogs and cats
Y. Takeuchi; K. Uetsuka; M. Murayama; F. Kikuta; A. Takashima; K. Doi; H. Nakayama, Neprilysin is an amyloid-beta-degrading enzyme localized in the brain parenchyma. The involvement of neprilysin in the pathogenesis of Alzheimer's disease has recently received much attention. We examined the localization of neprilysin and amyloid-beta, as well as the activity of neprilysin, in the brains of dogs and cats of various ages to clarify the relationship between neprilysin activity and amyloid-P deposition. The distribution of neprilysin was almost identical in dogs and cats, being high in the striatum, globus pallidus, and substantia nigra, but very low in the cerebral cortex. The white matter and hippocampus were negative. Neprilysin activity in the brain regions in dogs and cats was ranked from high to low as follows: thalamus/striatum > cerebral cortex > hippocampus > white matter. Amyloid-P deposition was first detected at 7 and 10 years of age in dogs and cats, respectively, and both the quantity and frequency of deposition increased with age. In both species, amyloid-P deposition appeared in the cerebral cortex and the hippocampus. In summary, the localization of neprilysin and neprilysin activity, and that of amyloid-beta, were complementary in the brains of dogs and cats., AMER COLL VET PATHOLOGIST
VETERINARY PATHOLOGY, 2008年07月, ［査読有り］

Intracranial meningioma with polygonal granular cell appearance in a Chihuahua
Yoshinori Takeuchi; Yumi Ohnish; Satoru Matsunaga; Hiroyuki Nakayama; Koji Uetsuka, A menigioma with polygonal granular cell proliferation in an 11-year and 8-month-old male Chihuahua is described. The tumor was observed under the dura matter of the right cerebrum. Microscopically, the tumor consisted of solid growth foci of small- or large-sized polygonal cells, with pale-stained nuclei, prominent nucleoli, and fine granular to foamy eosinophilic cytoplasm. Some of the proliferating cells contained variable amounts of cytoplasmic PAS-positive granules. Immunohistochemical analysis revealed that neoplastic cells were positive for vimentin and S-100 protein. Ultrastructurally, the neoplastic cells contained vesicular structures with a few small round-shaped bodies in the cytoplasm. We diagnosed the case as canine meningioma with granular cell appearance., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2008年05月, ［査読有り］

Neurotoxicity of MPTP to migrating neuroblasts: Studies in acute and subacute mouse models of Parkinson’s disease.
He XJ; Yamauchi H; Uetsuka K; Nakayama H, The acute or subacute administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) has been widely used in C57BL/6 mice to develop models of Parkinson's disease (PD). The loss of dopaminergic neurons is suggested to be mediated by a mechanism of nonapoptotic cell death or by apoptosis. In recent years, the notion that the neurotoxicity of MPTP is restricted to dopaminergic neurons in the substantia nigra (SN) has been challenged. Here, we provide evidence of rapid cell death in the subventricular zone (SVZ) and rostral migratory stream (RMS) in the adult C57BL/6 mouse brain in response to acute or subacute treatment with MPTP. Significant terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) of fragmented DNA was observed at 24 h (or 1 day) after the last injection in the acute model or after the first injection in the subacute model. Ultrastructural analysis confirmed that dying cells displayed an apoptotic morphology. Using a double labeling method, we demonstrated that the phenotype of the cells undergoing apoptosis is that of migrating neuroblasts. This is further supported by evidence of a subsequent loss of migrating neuroblasts. The results raise the possibility that migrating neuroblasts in the SVZ and RMS may be more vulnerable to MPTP than nigrostriatal dopaminergic neurons in the SN, and the death of migrating neuroblasts may be a primary event in the mouse model of PD. Furthermore, our data suggests that the death and subsequent loss of migrating neuroblasts in the acute or subacute model probably lead to a decreased potential for neurogenesis to some extent. (C) 2008 Elsevier Inc. All rights reserved., ELSEVIER SCIENCE BV
Neurotoxicology., 2008年05月, ［査読有り］

Relation between lipogranuloma formation and fibrosis, and the origin of brown pigments in lipogranuloma of the canine liver.　　　　　　　　　　　　　　　
Isobe K; Nakayama H; Uetsuka K
Comparative Hepatology., 2008年, ［査読有り］

Expression of epidermal growth factor receptor protein in the liver of db/db mice after partial hepatectomy
Makoto Shirai; Hirofumi Yamauchi; Hiroyuki Nakayama; Kunio Doi; Koji Uetsuka, Liver regeneration was impaired after partial hepatectomy (PH) in leptin receptor-deficient db/db mice with severe liver steatosis. In the present study, we analyzed the mode of epidermal growth factor receptor (EGFR) protein expression in the liver of 5- and 10-week-old db/db and age-matched control mice. In 5-week-old db/db mice, neither the expression of EGFR protein in the intact liver nor the rate of liver regeneration after PH was significantly different from that in age-matched control mice. However, in 10-week-old db/db mice, the level of EGFR protein expression was very low and liver regeneration was prominently suppressed. Histopathologically, much severer fatty change was observed in the liver of 10-week-old db/db mice than 5-week-old db/db mice. These results suggest that the down-regulation of EGFR protein expression is associated with an impairment of liver regeneration in db/db mice and that the severity of hepatic steatosis plays an indirect role in the impairment of liver regeneration by modifying EGFR expression. (c) 2007 Elsevier GmbH. All rights reserved., ELSEVIER GMBH, URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2007年11月, ［査読有り］

Essential role of p53 in trophoblastic apoptosis induced in the developing rodent placenta by treatment with a DNA-damaging agent
Hirofumi Yamauchi; Kei-ichi Katayama; Masaki Ueno; Xi Jun He; Takashi Mikami; Koji Uetsuka; Kunio Doi; Hiroyuki Nakayama, Placental apoptosis plays important roles in both normal morphogenesis and pathogenesis. We previously reported that administration of cytosine arabinoside (Ara-C), a DNA-damaging agent, to pregnant rats induced apoptosis of trophoblasts in the placental labyrinth zone. Our aim here was to clarify the molecular pathway of DNA damage induced-trophoblastic apoptosis. We found the accumulation and phosphorylation of p53 protein, a tumor suppressor that mediates apoptosis under various cellular stresses, in Ara-C-treated rat placentas. Expression of the mRNAs of downstream targets of p53 was upregulated, suggesting that p53 exerts its function as a transcription factor. We also observed release of mitochondrial cytochrome c and activation of caspase-9, hallmarks of the intrinsic apoptotic pathway. Phosphorylation of Chk1 and H2A.X, target substrates of DNA damage transducers, was detected immediately after Ara-C treatment, suggesting activation of DNA damage cascades to phosphorylate p53. Ara-C-induced trophoblastic apoptosis was almost completely abrogated in placentas of Trp53 (coding p53)deficient mice, whereas the levels of physiological apoptosis in trophoblasts were similar among wild-type and Trp53-deficient mice. These results indicate that p53 is essential for DNA damage-induced trophoblastic apoptosis and suggest that the mechanisms that regulate the damage-induced apoptosis differ from those that regulate physiological apoptosis., SPRINGER
APOPTOSIS, 2007年10月, ［査読有り］

Involvement of sex, strain and age factors in high fat diet-induced obesity in C57BL6J and BALB/cA mice
Satomi Nishikawa; Akira Yasoshima; Kunio Doi; Hiroyuki Nakayama; Koji Uetsuka, Although a number of obesity animal models have been reported, each model possesses different characteristics of obesity, suggesting care should be taken in choosing an animal model suitable for the experimental purpose. In this report, we fed 4-(young) and 52-week-old (middle-aged) C57BL/6J (B6) and young BALB/cA (BALB/c) mice with a high fat diet (HFD) for 9 weeks, and investigated the clinical and histological characteristics of obesity. In BALB/c mice, males gained more body weight and body fat weight and had higher energy intake than females by HFD feeding. Comparing the effect of HFD feeding between the strains of mice, BALB/c male mice accumulated more hepatic lipid than B6 male mice. In addition, middle-aged B6 mice increased the ratio of fat to body weight and hepatic lipid accumulation more than young mice. In conclusion, the characteristics of obesity induced by HFD feeding were influenced by the sex, strain and age of mice. Sex steroid hormones, hepatic lipid metabolism and systemic metabolism might be involved in these factors. The basic data in this study will be useful for the development of animal models of high fat diet-induced obesity., INT PRESS EDITING CENTRE INC
EXPERIMENTAL ANIMALS, 2007年07月, ［査読有り］

Histopathological characteristics of Kupffer cells and ito cells in the porcine and bovine liver
Koji Uetsuka; Satomi Nishikawa; Kaori Isobe; Hiroyuki Nakayama, We previously reported that no Kupffer cells reacted with the antibody against lysozyme, and Ito cells contained a large cytoplasmic vacuole in the feline liver. In this report, we further examined the characteristics of porcine and bovine hepatic non-parenchymal cells. In the liver of both animals, Kupffer cells were positive for lysozyme, and cytoplasmic vacuoles in Ito cells were small. The histopathological characteristics of porcine and bovine hepatic non-parenchymal cells were different from those of the feline liver., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2007年07月, ［査読有り］

Vitamin K deficiency of germfree mice caused by feeding standard purified diet sterilized by gamma-irradiation
Kazuhiro Hirayama; Koji Uetsuka; Yoshibumi Kuwabara; Motoi Tamura; Kikuji Itoh, Germfree mice died when they were fed a purified diet of AIN-76 formula sterilized by gamma-irradiation. Vitamin K deficiency was suspected and this study was performed to confirm the cause of the death. Germfree mice were fed purified diets of AIN-76 or AIN-93M formula, which were pelleted and sterilized by gamma-irradiation at a dose of 50 kGy. One half of the mice fed the AIN-76 diet died within two weeks and the surviving animals were also in poor health, while 91% of mice fed the AIN-93M diet survived. No hemorrhage was observed grossly in any organs of the surviving animals. Histologically, degeneration with inflammatory cell infiltration was observed as well as hemorrhage and fibrosis in the heart muscles of mice fed the AIN-76 diet. No microscopic lesions were observed in the other organs. Prothrombin time (PT) and activated partial thromboplastin time (APTT) were extremely prolonged when mice were fed the AIN-76 diet. The animals totally recovered when they were intragastrically administered 1 mu g/day of vitamin K-3 from the third day of feeding of the AIN-76 diet, except for PT and APTT which were still slightly longer than in mice fed the AIN-93M diet. The concentration of vitamin K3 supplied in the AIN-76 diet decreased to an undetectable level after gamma-irradiation, while the AIN-93M diet contained 240 mu g/kg of vitamin K-1. These results indicate that the deaths of the germfree mice fed the gamma-irradiated AIN-76 diet were caused by vitamin K deficiency. Vitamin K deficiency may cause fatal degeneration of cardiac muscle cells., INT PRESS EDITING CENTRE INC
EXPERIMENTAL ANIMALS, 2007年07月, ［査読有り］

Measurement of intestinal mucosal permeability in dogs with lymphocytic-plasmacytic enteritis
Saku Kobayashi; Koichi Ohno; Koji Uetsuka; Ko Nakashima; Asuka Setoguchi; Yasuhito Fujino; Hajime Tsujimoto, Lymphocytic-plasmacytic enteritis (LPE) is a type of canine inflammatory bowel disease (IBD). One of its most probable causes is a defect in the mucosal permeability barrier. In the present study, intestinal permeability in LPE dogs was examinated to evaluate its clinical value. Twenty-nine dogs with LPE diagnosed by clinical and histological examinations were included in this study. Intestinal permeability was evaluated by measuring the ratio of the concentrations of two sugars (lactulose (L) and rhamnose (R)) with different molecular weights in urine samples after oral administration of a solution containing them. Biopsy specimens of duodenum were evaluated according to histological criteria. The urinary L:R ratio in the 29 LPE dogs (1.68 +/- 1.17, mean +/- SD) was significantly higher than that in the 10 healthy control dogs (0.75 +/- 0.38, P < 0.01). In the LPE dogs, a weak correlation was observed between the histopathological grading score of the duodenum and the urinary L:R ratio (r=0.408, P < 0.05). The urinary L:R ratio in the 20 dogs showing hypoalbuminemia (< 2.5 g/dl) was significantly higher than that in the 9 dogs with normal serum albumin levels > 2.5 g/dl (P < 0.01). In conclusion, permeability of the intestinal mucosa as determined by the urinary L:R ratio could be a useful laboratory parameter for evaluating intestinal damage in LPE dogs., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2007年07月, ［査読有り］

Histopathological characteristics of hepatic lipogranulomas with portosystemic shunt in dogs.　　　　　　　　　　　　　　　
Isobe K; Matsunaga S; Nakayama H; Uetsuka K
The Journal of Veterinary Medical Science., 2007年

Thymic granulomatous lesions in pigs
Y. Baba; H. Nakayama; A. Yasoshima; K. Uetsuka; S. Kumagai; K. Doi, Rare cases of thymic granulomatous lesions were found in pigs. The lesions consisted of epithelioid cells, multinucleated giant cells, and lymphocytes. Such lesions also were observed in the mesenteric lymph nodes, spleen, kidney, and stomach. The cytoplasm of the majority of giant cells and some epithliold cells was periodic acid-Schiff (PAS) positive. All cells were positive for vimentin, lysozyme, and desmin. Ultrastructurally, the giant cells were rich in organella and attached to adjacent epithelioid cells by membrane interdigitation. The cells included numerous coated vesicles and granules. No etiologic pathogen, including porcine circovirus type 2, was detected in the lesions. This is the rare case of idiopathic thymic granulomatous lesion in pigs., AMER COLL VET PATHOLOGIST
VETERINARY PATHOLOGY, 2006年11月, ［査読有り］

Evidence of apoptosis in the subventricular zone and rostral migratory stream in the MPTP mouse model of Parkinson disease
Xi Jun He; Hiroyuki Nakayama; Mei Dong; Hirofumi Yamauchi; Masaki Ueno; Koji Uetsuka; Kunio Doi, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is commonly used to create animal models of Parkinson disease. There is conflicting evidence on the occurrence of apoptosis induced by MPTP in the mouse substantia nigra pars compacta. We demonstrated that a single acute injection of MPTP induced apoptosis in the subventricular zone (SVZ) and rostral migratory stream (RMS) in the adult C57BL/6 mouse brain. The number of TUNEL-positive cells peaked at 24 hours after injection and decreased thereafter, paralleling the change in the number of cleaved caspase-3-positive cells after MPTP injection. Results of immunohistochemistry and ultrastructural analyses indicated that the majority of apoptotic cells in the SVZ and RMS were migrating neuroblasts (type A cells), whereas a few were astrocytes (type B cells). No apoptosis occurred in transit-amplifying progenitors (type C cells). The decrease in A cell numbers was most marked on day 2 and lasted to day 8 after the administration. A rapid and transient phagocytosis of apoptotic cells by microglial cells was demonstrated to parallel the MPTP-induced apoptosis. The present findings provide new insight into the extensive neurotoxicity of MPTP and may be valuable in reevaluating the MPTP mouse model of Parkinson disease., LIPPINCOTT WILLIAMS & WILKINS
JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY, 2006年09月, ［査読有り］

Encepalomyocarditis virus-induced apoptosis and ultrastructural changes in the lacrimal and parotid glands of mice
A Ohguchi; Y Nakayama; A Yasoshima; C Doi; T Mikami; K Uetsuka; H Nakayama; K Doi, Development of acinar cell apoptosis and ultrastructural changes in the exorbital lacrimal and parotid glands was examined in DBA/2 mice infected with 10(2) PFU/mouse of EMC-D virus. Pyknotic acinar cells, most of which were positive for TUNEL and cleaved caspase-3 and had ultrastructural characteristics of apoptotic cells, developed earlier and were more frequently observed in the parotid gland than in the exorbital lacrimal gland, while the total damage of acinar cells and interstitial infiltration of macrophages were more prominent in the latter than in the former. These findings indicate that EMC-D virus induces acinar cell apoptosis in these glands. In addition, corresponding to the results of the detection of viral RNA signals by in situ hybridization, small aggregates of virus-like particles having typical size and structure of EMC virus were frequently observed in both the cytoplasm and the nucleus of acinar cells in the exorbital lacrimal gland, while they were found only in the cytoplasm of a few acinar cells in the parotid gland. In conclusion, between the exorbital lacrimal and parotid glands, there was a reverse relationship observed between the development of acinar cell apoptosis and that of total damage of acinar cells. (c) 2005 Elsevier Inc. All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
EXPERIMENTAL AND MOLECULAR PATHOLOGY, 2006年04月, ［査読有り］

Histopathological characteristics of Ito cells and Kupffer cells in the feline liver
K Uetsuka; S Nishikawa; A Yasoshima; H Nakayama; K Doi, The histopathological characteristics of Ito cells and Kupffer cells were investigated in the liver of 21 cats (age range: 6 months -18 years) autopsied in our laboratory during 2003. Immunohistochemical examinations were performed using antibodies against lysozyme, desmin and alpha-smooth muscle actin. No Kupffer cells reacted with the antibody against lysozyme. However, macrophages in the lung and spleen showed a positive reaction with the antibody. This finding suggests a possibility that the amount of lysozyme in the Kupffer cells of feline liver is comparatively small. On the other hand, large vacuole-laden cells were observed in the hepatic perisinusoid of some feline cases, and these cells showed a positive reaction with antibodies against desmin and alpha-smooth muscle actin. These cells could be Ito cells with large lipid vacuoles. This conclusion was supported by electron microscopic observation and oil red O staining. However, no such large vacuole-laden perisinusoidal cells were detected in the liver of young cats less than 2 years old. The present study revealed the histopathological features of Kupffer cells and Ito cells in the feline liver., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2006年03月, ［査読有り］

Histopathological changes in the brain of mouse fetuses by etoposide-administration
C Nam; GH Woo; K Uetsuka; H Nakayama; K Doi, Etoposide (VP-16), a topoisomerase II inhibitor, is an anti-tumor agent which is also known to show embryotoxicity, and teratogenicity when administered to pregnant rodents. We examined VP-16-induced histopathological changes in the brain of mouse fetuses. Pregnant mice were intraperitoneally injected with VP-16 (4 mg/kg) on day 12 of gestation (GD12), and fetuses were collected from 1 to 48 hours after treatment (HAT). Mitotic neuroepithelial cells in the telencephalic wall prominently decreased at 2 HAT, and were hardly observed at 4 HAT. The number of pyknotic neuroepithelial cells in the fetal brain began to increase at 4 HAT, and became prominent from 8 to 24 HAT. These pyknotic cells were also positively stained by TUNEL method, which can detect fragmented DNA, and showed ultrastructural characteristics of apoptosis. Additionally, these cells were also positive for cleaved caspase-3, an essential executioner of apoptosis. This indicated that excessive neuroepithelial cell apoptosis was induced in the brain of mouse fetuses following VP16 treatment on GD 12., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 2006年03月, ［査読有り］

The activity of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in mammary tumors of dogs and rats
K Kawai; K Uetsuka; K Doi; H Nakayama, We conducted zymography for detecting the activity of matrix metalloproteinases (MMPs) and reverse zymography for the activity of tissue inhibitors of metalloproteinases (TIMPs) in canine spontaneous and rat 7, 12-dimethylbenz(a)anthracene (DMBA)induced mammary tumor tissues. The activities of MMPs of canine mammary tumors were quite higher than those of the rat chemically induced tumors. The activities of MMPs were significantly higher in malignant tissues than in benign ones of canine tumors, whereas the activity of only MMP-2 was higher in both benign and malignant rat tumors compared to normal tissues. There were no differences of MMPs activities between benign and malignant rat tumors. The results of reverse zymography indicated that the activities of TIMP-1, -2 and -3 were strikingly higher in rat tumors than in canine tumors. The activities were higher in malignant tissues than in benign ones of dogs, and higher in tumor tissues than in normal mammary tissues of rats. The results of film in situ zymography for tissue localization of gelatinolytic activity showed that the digested area was more extended in malignant tumors than in benign ones of dogs. However, the area was similarly extended in both benign and malignant rat tumors. These results may indicate that the canine spontaneous malignant mammary tumors possess more aggressive nature than the rat chemically induced counterpart, resulting from the high level of MMPs and low level of TIMPs activities of the tumor tissues., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2006年02月, ［査読有り］

乳管拡張症を伴ったフェレットの副腎皮質腺癌の１例．
三輪恭嗣、松永悟、上塚浩司、中山裕之、田中隆志、谷口孝、小川博之., 2年以上の全身性脱毛を呈していた5歳8カ月齢の避妊雌フェレットで右第3～5乳腺の腫大が確認された. エストラジオール, 17-ヒドロキシプロゲステロンの高値およびその他の臨床検査結果から脱毛は副腎疾患に関連した症状と判断した. 開腹により左右副腎の腫大を認めた. 左副腎および乳腺腫脹部の外科的切除と病理組織学的検査を実施し, それぞれ副腎皮質腺癌および乳管拡張症と診断した.術後, 術前に高値を示した血清中ステロイドホルモン値は低下し, 臨床経過から, 本症例は副腎腫瘍により過剰に分泌された性ステロイドホルモンの影響を乳腺が長期間受けた結果, 乳管拡張症が発生したものと推測された., Japan Veterinary Medical Association
日本獣医師会雑誌, 2006年, ［査読有り］

Two cases of feline malignant craniopharyngioma. (vol 42, pg 663, 2005)
T Nagata; H Nakayama; K Uchida; K Uetsuka; A Yasoshima; S Yasunaga; K Masuda; H Tsujimoto; E Kuwajima; R Nishimura; N Sasaki; K Doi, AMER COLL VET PATHOLOGIST
VETERINARY PATHOLOGY, 2005年11月, ［査読有り］

Two cases of feline malignant craniopharyngioma
T. Nagata; H. Nakayama; K. Uchida; K. Uetsuka; A. Yasoshima; S. Yasunaga; K. Masuda; H. Tsujimoto; E. Kuwajima; R. Nishimura; N. Sasaki; K. Doi, Tumors at the cranial base in 2 cats (a 91/2-year-old, castrated male Chinchilla and a 7-year-old, castrated male American Shorthair) were diagnosed as malignant craniopharyngioma. The tumor lesion was histopathologically divided into four parts: 1) a small acinus part, in which relatively large cells with a pale cytoplasm composed small acini
2) a duct part, in which small cuboidal cells composed ducts
3) a cyst part, in which there were large cysts lined with flat cells
and 4) a pavement part, in which large multiangular-shaped cells proliferated in a pavement pattern. The epithelial cells of some parts were positive for keratin by immunohistochemistry. Histopathologic findings of the present feline cases were identical to those of malignant craniopharyngioma in other animal species.
Veterinary Pathology, 2005年09月, ［査読有り］

Impaired proliferation of non-parenchymal cells participates in an impairment of liver regeneration in db/db mice
K Uetsuka; M Shirai; H Yamauchi; H Nakayama; K Doi, In this study, we examined the possibility that impaired proliferation of non-parenchymal cells affects in an impairment of liver regeneration in db/db mice, which are congenitally deficient in receptors for leptin. Liver regeneration after a two thirds partial hepatectomy (2/3 PH) was impaired in 10-week-old female db/db mice. The proliferation of both hepatocytes and non-parenchymal cells estimated from a bromodeoxyuridine (BrdU) labeling index was suppressed, and the protein expression of vascular endothelial growth factor was blocked in db/db mice. Although the extent of fatty change and the level of epidermal growth factor receptor protein expression in the liver were improved in 5-week-old db/db mice, the regeneration of liver was impaired after 2/3 PH in both 5- and 10-week-old db/db mice. These results suggested that suppressed proliferation of non-parenchymal cells contributes to the impairment of liver regeneration in db/db mice. As leptin has also the angiogenic effect, the angiogenic inhibitor FR-118487 was administered to ICR mice to examine liver regeneration after 2/3 PH, and the rate of regeneration was affected. In conclusion, it is suggested that the Suppressed proliferation of non-parenchymal cells contributes to the impairment of liver regeneration probably through a disrupted angiogenesis in db/db mice. (C) 2005 Elsevier Inc. All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
EXPERIMENTAL AND MOLECULAR PATHOLOGY, 2005年08月, ［査読有り］

Microarray analysis of T-2 toxin-induced liver, placenta and fetal liver lesions in pregnant rats
S Sehata; N Kiyosawa; F Atsumi; K Ito; T Yamoto; M Teranishi; K Uetsuka; H Nakayama; K Doi, Pregnant rats on day 13 of gestation were treated orally with 2 mg/kg of T-2 toxin and sacrificed at 1, 3, 6, 9 and 12 It after the treatment (HAT). Histopathologically, the number of apoptotic cells was increased in the liver, placenta and fetal liver (peaked at 6, 12 and 9-12 HAT, respectively). To examine the gene expression profiles, we performed microarray analysis of these tissues at two selected time points based on the results of the TdT-mediated dUTP nick end labeling (TUNEL) staining. Increased expression of oxidative stress- and apoptosis-related genes was detected in the liver of dams, placenta and fetal liver of pregnant rats treated with T-2 toxin at the peak time point of apoptosis. Decreased expression of lipid metabolism- and drug-metabolizing enzyme-related genes was also detected in these tissues. The results suggested that the mitogen-activated protein kinase (MAPK) pathway might be involved in the mechanism of T-2 toxin-induced apoptosis. In addition, increased expression of the c-jun gene was consistently observed in these tissues. Our results suggest that the mechanism of T-2 toxin-induced toxicity in pregnant rats is due to oxidative stress followed by the activation of the MAPK pathway, finally inducing apoptosis. The c-jun gene may play an important role in T-2 toxin-induced apoptosis. (C) 2005 Elsevier GmbH. All rights reserved., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2005年08月, ［査読有り］

アリューシャン病ウイルスフェレット由来株に感染し斃死したフェレットの１例．　　　　　　　　　　　　　　　
三輪恭嗣、松永悟、中山裕之、上塚浩司、中村裕之、小川博之.
日本獣医師会雑誌, 2005年, ［査読有り］

G(M2)-gangliosidosis variant 0 (Sandhoff-like disease) in a family of Japanese domestic cats
O Yamato; S Matsunaga; K Takata; K Uetsuka; H Satoh; T Shoda; Y Baba; A Yasoshima; K Kato; K Takahashi; M Yamasaki; H Nakayama; K Doi; Y Maede; H Ogawa, A five-month-old, female Japanese domestic shorthair cat with proportionate dwarfism developed neurological disorders, including ataxia, decreased postural responses and generalised body and head tremors, at between two and five months of age. Leucocytosis due to lymphocytosis with abnormal cytoplasmic vacuolations was observed. The concentration of G(M2)-ganglioside in its cerebrospinal fluid was markedly higher than in normal cats, and the activities of beta-hexosaminidases A and B in its leucocytes were markedly reduced. on the basis of these biochemical data, the cat was diagnosed antemortem with G(M2)-gangliosidosis variant 0 (Sandhoff-like disease). The neurological signs became more severe and the cat died at 10 months of age. Histopathologically, neurons throughout the central nervous system were distended, and an ultrastructural study revealed membranous cytoplasmic bodies in these distended neurons. The compound which accumulated in the brain was identified as G(M2)-ganglioside, confirming G(M2)-gangliosidosis. A family study revealed that there were probable heterozygous carriers in which the activities of leucocyte beta-hexosaminidases A and B were less than half the normal value. The Sandhoff-like disease observed in this family of Japanese domestic cats is the first occurrence reported in Japan., BRITISH VETERINARY ASSOC
VETERINARY RECORD, 2004年12月, ［査読有り］

Morphological and microarray analysis of T-2 toxin-induced rat fetal brain lesion
S Sehata; N Kiyosawa; T Makino; F Atsumi; K Ito; T Yamoto; M Teranishi; Y Baba; K Uetsuka; H Nakayama; K Doi, To examine morphological and gene expression changes induced by T-2 toxin in the fetal brain in detail, pregnant rats on day 13 of gestation were treated orally with a single dose of T-2 toxin (2 mg/kg) and sacrificed at 1, 3, 6, 9, 12 and 24 h after treatment (HAT). Histopathologically, the number of apoptotic neuroepithelial cells in the telencephalon increased from 1 HAT and peaked at 12 HAT. Based on the histopathological examinations, microarray analysis was performed at 6, 12 and 24 HAT. Microarray analysis showed that the expression of oxidative stress-related genes (heat shock protein 70 (HSP70) and heme oxygenase (HO)) was strongly induced by T-2 toxin at 12 HAT, the peak time point of apoptosis induction. The expression of mitogen-activated protein kinase (MAPK)-related genes (MEKK1 and c-jun) and other apoptosis-related genes (caspase-2 and insulin-like growth factor-binding protein-3 (IGF-BP3)) was also induced by the T-2 toxin treatment. The changes observed by microarray analysis were confirmed for four up-regulated genes (HSP70, HO, IGF-BP3 and VEGF-A) using real-time RT-PCR. Our results suggest that the T-2 toxin-induced apoptosis in the fetal brain is due to oxidative stress, and that the MAPK pathway may be involved in T-2 toxin-induced toxicity. (C) 2004 Elsevier Ltd. All rights reserved., PERGAMON-ELSEVIER SCIENCE LTD
FOOD AND CHEMICAL TOXICOLOGY, 2004年11月, ［査読有り］

Effects of prenatal hydroxyurea-treatment on mouse offspring
GH Woo; K Katayama; EJ Bak; M Ueno; H Yamauchi; K Uetsuka; H Nakayama; K Doi, Hlydroxyurea (HU), a ribonucleotide reductase inhibitor, induces morphological anomalies in the central nervous system, craniofacial tissues and limb buds in animals, and neonatal respiratory distress in humans. The neonates and offspring of pregnant mice treated with 400 or 800 mg/kg of HU on day 13 of gestation were examined at 0 day and 10 weeks after birth to find a clue for clarifying the relationship between HU-induced apoptosis in the fetal tissues and teratogenicity. The offspring from dams treated with HU were retarded in growth compared with controls. But there was no significant difference in the body weight gain between the 400 and 800 mg/kg groups. In the teratologic changes, microencephaly, hydrocephalus and curved coccygeal vertebrae were observed in the offspring, and the incidence of these teratologic changes was similar but their degree was more severe in the 800 mg/kg group than in the 400 mg/kg group. Based on the above-mentioned previous and present studies of ours, we suggest that HU-induced apoptosis in fetal tissues may play an important role in the development of anomalies in the corresponding tissues of offspring. (C) 2004 Elsevier GmbH. All rights reserved., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2004年10月, ［査読有り］

Involvement of p53 in 1-beta-D-arabinofuranosylcytosine-induced rat fetal brain lesions
H Yamauchi; K Katayama; M Ueno; K Uetsuka; H Nakayama; K Doi, 1-beta-D-Arabiiioftiranosylcytosine (Ara-C), a cytidine analogue cytotoxic to proliferating cells, has a teratogenic effect in the brain of experimental animals and causes neural cell apoptosis in vitro and in vivo. In the present study, pregnant rats were injected with Ara-C on Day 13 of gestation and the fetal brain was collected from 1 to 48 It after treatment. Histopathological examinations revealed marked induction of apoptotic cell death and decrease of mitosis in neuroepithelial cells in the brain of Ara-C-treated fetus, and these changes were most prominent from 9 to 12 h. Expression of p53 protein, which mediates apoptosis and cell cycle arrest after DNA damage, was elevated remarkably and peaked at 3 h. p21, a cyclin-dependent kinase inhibitor responsible for p53-mediated cell cycle arrest, showed intense overexpression in protein and mRNA levels following the increase of p53 protein. The mRNA expressions of other p53 transcriptional target genes, bax, cyclinG1, and fas, also significantly increased and peaked at around 9 h. In conclusion, prenatal treatment of Ara-C is thought to induce apoptosis and inhibition of cell proliferation mediated by p53 and its target genes in the fetal brain. (C) 2004 Elsevier Inc. All rights reserved., PERGAMON-ELSEVIER SCIENCE LTD
NEUROTOXICOLOGY AND TERATOLOGY, 2004年07月, ［査読有り］

Thymoma in a dog with a part of granular cell proliferation and concurrent lymphoma cells
A Fukuoka; H Nakayama; Y Nakayama; A Yasoshima; K Uetsuka; Y Fujino; H Tsujimoto; K Doi, A 12-year-old male Shiba dog showed anemia and the swelling of systemic lymph nodes. X-ray and post mortal examinations revealed a anterior mediastinal mass. Histologlcally, the tumor mass consisted of four different elements; cord-like proliferation of cuboidal epithelial cells, tubular or cystic structures lined with ciliated epithelial cells, proliferation of large round-shaped epithelial cells with PAS-slightly positive granular cytoplasm, and diffuse proliferation of neoplastic lymphocytes. Epithelial cells in cord-like or cystic structures were strongly positive for cytokeratin. Granular or foamy cells were negative for all markers examined and had myelin-like bodies in the cytoplasm by electron microscopy. The neoplastic lymphocytes in the tumor mass were considered being derived from concurrent multicentric lymphoma. Based on these findings, the present case was diagnosed as thymoma with a part of granular cell proliferation and concurrent lymphoma cells., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2004年06月, ［査読有り］

Involvement of p53 in 1-beta-D-arabinofuranosylcytosine-induced trophoblastic cell apoptosis and impaired proliferation in rat placenta
H Yamauchi; K Katayama; M Ueno; K Uetsuka; H Nakayama; K Doi, 1-beta-D-Arabinofuranosyicytosine (Ara-C), a DNA-damaging agent, severely inhibits fetal growth and has teratogenicity. Recently, we reported that Ara-C also causes placental growth retardation and increases placental apoptosis. The aim of the present study is to elucidate the mechanisms of placental injury induced by genotoxic stress and involvement of p53, which mediates apoptosis and cell-cycle arrest after DNA damage. We injected Ara-C into pregnant rats on Day 13 of gestation and examined the placentas from 1 to 48 h after the administration. Terminal deoxynucleotidyltransferase-mediated dUTP end-labeling (TUNEL) revealed that the apoptosis of trophoblastic cells in the placental labyrinth zone increased from 3 h after the treatment and peaked at 6 h before returning to control levels at 48 h. An increase in cleaved caspase-3 immunoreactivity was also detected at 6 h. Proliferative activity as measured by immunohistochemistry for topoisomerase IIalpha and by mitotic index significantly decreased after the treatment in the labyrinth zone. Immunoreactivity for p53 protein in the placental labyrinth zone was remarkably enhanced and peaked at 3 h after treatment, although no increase in p53 mRNA expression was detected with a reverse transcription-polymerase chain reaction. Regarding p53 target genes, p21, cyclinG1, and fas mRNA levels increased significantly and peaked at around 9 h after the treatment. These results indicate that Ara-C would induce apoptosis and impair cell proliferation in the placental labyrinth zone, and p53 and its transcriptional target genes may play an important role in the pathogenesis of the Ara-C-induced placental toxicity., SOC STUDY REPRODUCTION
BIOLOGY OF REPRODUCTION, 2004年06月, ［査読有り］

Rat strain differences in the early stage of porcine-serum-induced hepatic fibrosis
Y Baba; K Uetsuka; H Nakayama; K Doi, Rat strain differences in the early development of porcine serum (PS)-induced hepatic fibrosis were histologically and immunohistochemically examined using Brown Norway (BN), Sprague Dawley (SD) and Wistar rats. They were injected i.p. with 0.5ml sterile PS twice a week for and 8 weeks. In addition. rats treated with physiological saline in the same way served as controls. At 4 weeks, hepatic fibrosis accompanying fibrous septa mainly composed of type III collagens developed in BN and SD rats but not in Wistar rats. In addition, the numbers of eosinophils, CD3-positive cells and ED-1-positive cells significantly increased in BN and SD rats, that of CD45RA-positive cells in BN rats. and that of alpha-smooth muscle actin (SMA)-positive cells in SD rats. respectively. Such differences in the number of inflammatory cells may be related with the absence of hepatic fibrosis in Wistar rats at 4 weeks. At 8 weeks, hepatic fibrosis with formation of many small-sized pseudolobules was observed in all strains at almost similar degree. and the numbers of infiltrating cells increased in all strains of rats with some exception. In addition, the main location of inflammatory cells was different. suggesting a different role of each inflammatory cell in the process of hepatic fibrosis., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2004年03月, ［査読有り］

Gene expression profiles in pregnant rats treated with T-2 toxin
S Sehata; N Kiyosawa; K Sakuma; K Ito; T Yamoto; M Teranishi; K Uetsuka; H Nakayama; K Doi, Pregnant rats on day 13 of gestation were treated orally with T-2 toxin at a single dose of 2 mg/kg and sacrificed at 24 hours after treatment. Histopathologically, apoptosis was increased in the liver. placenta and fetal liver. Microarray analysis was performed to examine the gene expression in the liver, placenta, and fetal liver. The results of microarray analysis showed that the changes in the expression of apoptosis genes. metabolic enzymes and oxidative stress-related genes were detected in these tissues. Suppression of phase I and II enzymes-related genes expression in the liver, and suppression of phase II enzymes-related genes expression in the placenta and fetal liver were observed. Semiquantitive RT-PCR analysis also showed the same results as those of microarray analysis. From the results of microarray analysis and histopathological examination, T-2 toxin seems to induce oxidative stress in these tissues, following the chances in metabolism-related genes expression. These changes may alter the intracellular environments resulting in the induction of apoptosis. Further studies on the gene expression profiles at the earlier time point are necessary to clarify the detailed mechanisms of T-2 toxin-induced toxicity in pregnant rats., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2004年03月, ［査読有り］

Effects of granulocyte colony-stimulating factor (G-CSF) on bleomycin-induced lung injury of varying severity
K Adachi; M Suzuki; T Sugimoto; K Yorozu; H Takai; K Uetsuka; H Nakayama; K Doi, We evaluated the effects of granulocyte colony-stimulating factor (G-CSF) on bleomycin (BLM)-induced lung injury that developed diffuse alveolar damage and subsequent pulmonary fibrosis (PF) of varying severity. G-CSF (100 mug/kg/day) was administered subcutaneously to BLM (0.2, 20, 2,000 mug)-treated or -untreated rats for 3 or 14 days. In the BLM 0.2 mug group, slight alveolar mononuclear cell infiltration was observed, although PF was not noted. In the BLM 20-mug and 2,000-mug groups, diffuse alveolar damage along with neutrophil infiltration and subsequent PF were observed. In the saline + G-CSF group and BLM 0.2 mug + G-CSF group, a marked increase in the number of alkaline phosphatase (ALP)-positive neutrophils was noted in the alveolar capillaries, although there was neither neutrophil infiltration in alveoli nor exacerbation of lung injury. In the BLM 20 mug + G-CSF and BLM 2,000 mug + G-CSF groups, an exacerbation of lung injury along with an increase in the number of ALP-positive neutrophils in the alveoli was observed. These results indicate that the administration of G-CSF to rats with slight lung injury bearing no PF does not exacerbate the lung injury. The exacerbating effects of G-CSF seem to be associated not only with the marked infiltration of activated neutrophils but also with the severity of underlying lung injury., TAYLOR & FRANCIS INC
TOXICOLOGIC PATHOLOGY, 2003年11月, ［査読有り］

イヌ乳腺腫瘍における遺伝子発現の網羅的解析　　　　　　　　　　　　　　　
川井 康司; 上塚 浩司; 片山 圭一; 中川 貴之; 佐々木 伸雄; 中山 裕之; 土井 邦雄, (公社)日本獣医学会
日本獣医学会学術集会講演要旨集, 2003年09月

Ultrastructural changes in the dorsal skin of Wistar-derived hypotrichotic WBN/ILA-Ht rats following UVA-irradiation
T Okada; A Yasoshima; K Uetsuka; H Nakayama; K Doi, Ultrastructual characteristics of the dorsal skin responses to a single irradiation of UVA (1100 kJ/m(2)) were examined in Wistar-derived hypotichotic WBN/ILA-Ht rats (HtRs). In the epidermis, mitochondrial swelling of some keratinocytes and dissociation of keratinocytes due to intercellular edema developed at 3 hours (h) after irradiation and continued to 48 h. At 6 h, in addition to these changes, necrosis of keratinocytes accompanied with infiltration of neutrophils was also observed in some portions, and epidermal hyperplasia with many keratinocytes showing nucleolar hypertrophy and some mitotic keratinocytes was observed at 48 h. In the dermis, mitochondrial swelling and/or partial cytoplasmic destruction in capillary endothelial cells and edema with inflammatory cell infiltration were observed at and after 3 h, and extravasation of erythrocytes was found in some capillaries at 48 h. Mitochondrial swelling was also frequently found in pericytes and fibroblasts. Inflammatory cells were mainly composed of neutrophils throughout the experimental period. Mild degranulation of mast cells which also showed mitochondrial swelling was observed at and after 3 h, and a close special relationship between mast cells and fibroblasts or neutrophils was sometimes observed. In conclusion, the most prominent change in the dorsal skin of HtRs exposed to UVA was degeneration of capillary endothelial cells, resulting, in edema and inflammatory cell infiltration, and the most characteristic cytopathic effect of UVA was mitochondrial swelling, and it was common to keratinocytes, capillary endothelial cells, pericytes, mast cells, and fibroblasts., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2003年09月, ［査読有り］

Distribution of N-methyl-D-aspartate receptors (NMDARs) in the developing rat brain
H Takai; K Katayama; K Uetsuka; H Nakayama; K Doi, The N-methyl-D-aspartate receptor (NMDAR), which is one of the glutamate receptors, is considered to have a close relationship to synaptic plasticity in the developing brain. In addition, it is known that the excessive stimulation of NMDARs can trigger neuronal apoptosis. In this study, we examined the distribution of NMDAR subunits [anti-NR1, NR2(A-C)] in the developing rat brain immunohistochemically. As a result, NR1, an essential subunit for the formation of a functional NMDAR complex, was mildly expressed ill the restricted areas such as the temporal region of the cerebral cortex and the hippocampus in the fetal brain at Embryonal Days 18 and 20. On the other hand. in neonates, NR1 was expressed widely throughout the whole brain. The distributions of NR2A and NR2C showed temporal and spatial similarities to that of NR1, while the expression of NR2B showed differences in the intensity and distribution. A progressive change in subunit expression seen prenatally and postnatally could contribute to variation of NMDARs and synaptic plasticity during the developing period. (C) 2003 Elsevier Science (USA). All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
EXPERIMENTAL AND MOLECULAR PATHOLOGY, 2003年08月, ［査読有り］

Effects of granulocyte colony-stimulating factor on the kinetics of inflammatory cells in the peripheral blood and pulmonary lesions during the development of bleomycin-induced lung injury in rats
K Adachi; M Suzuki; T Sugimoto; K Uetsuka; H Nakamaya; K Doi, We evaluated the effects of granulocyte colony-stimulating factor (G-CSF) on the kinetics of inflammatory cells during the development of inflammation in bleomycin (BLM)-induced lung injury. G-CSF (100 mug/kg/day, s.c.) was administered to rats treated with or without BLM (2 mg/200 mul, intratracheally) for up to 14 days (Day 14) immediately after BLM treatment. In the BLM + G-CSF group, the lung injury score increased on Days I and 14, and the score of lung fibrosis on Day 14, respectively. Except for neutrophils, there were no effects of G-CSF on the number of inflammatory cells both in the peripheral blood and in the lung in both BLM-treated and -untreated rats at the acute inflammatory phase. In the G-CSF-treated groups, the number of neutrophil counts in the peripheral blood drastically increased on Day 1, temporally decreased on Day 3, and increased again on Days 7 and 14. The number of neutrophils in the lung markedly increased on Day 1 and then remained at a plateau level until Day 14. The neutrophil alkaline phosphatase score in the lung commenced to increase on Day 1, reached the maximal level on Day 7, and then remained at a plateau level until Day 14. Correlations between the numbers of neutrophils in the lung and the peripheral blood or the lung lesion score were only observed on Day 14. These findings suggest that the exacerbating effect of G-CSF on the lung injury coincided with the increase in the number of alkaline phosphatase-positive neutrophils infiltrating in the pulmonary lesion at the acute inflammatory phase and it lasted to the fibrogenic phase. The exacerbating effect of G-CSF on the severe BLM-induced lung injury seems to be related not only to the pulmonary accumulation of activated neutrophils but also to the severity of lung injury caused by the direct effects of BLM., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2003年07月, ［査読有り］

Sunburn reaction in the dorsal skin of hypotrichotic WBN/ILA-Ht rats
T Okada; SM Alberenque; A Yasoshima; Malcotti, V; K Katayama; K Uetsuka; H Nakayama; K Doi, The dorsal skin responses to a 2 single irradiation with a high-dose of UVB (10kJ/m(2)) were examined histologically and immunohistochemically in UVB-sensitive Wistar-derived hypotrichotic WBN/ILA-Ht rats (HtRs). Sunburn cells (SBCs) which were characterized by pyknotic nuclei and eosinophilic cytoplasm and had ultrastructual characteristics of apoptotic cells were first observed in the epidermis at 3 hours (h) after irradiation. The number peaked at 6 h, and then decreased rapidly. The expressions of p53 protein, which is known to be closely related to the formation of SBCs, and of p21 protein, which is one of the transcriptional target genes of p53, were immunohistochemically detected, and their labeling index (LI) in the epidermis peaked at 12 to 24 It (p53) or at 24h (p21) after irradiation. On the other hand, proliferating cell nuclear antigen (PCNA)-LI in keratinocytes was significantly lower than the control group at 6 h after irradiation and thereafter it increased and became significantly higher than the control group from 24 to 48 It. At 48 h, moderate hyperplasia with moderate numbers of mitotic keratinocytes was first observed in the epidermis. In the dermis, mild edema developed from 12 to 36 h and it accompanied mild lymphocyte infiltration at 36 It. Judging from the present results, it was suggested that some factors other than p53 might be involved in SBC formation, and that p53 might induce p21 protein and play an important role in cell growth arrest in keratinocytes after UVB irradiation., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 2003年07月, ［査読有り］

NMDA-induced apoptosis in the developing rat brain
H Takai; K Katayama; A Yasoshima; K Uetsuka; H Nakayama; K Doi, The N-methyl-D-aspartate receptor (NMDAR), which is one of the glutamate receptors, is considered to have a close relation to synaptic plasticity in the developing brain. In addition, it is also known that the excessive stimulation of NMDARs can trigger neuronal apoptosis. In this study, we examined the expression of neuronal apoptosis in the developing rat brain after the administration of NMDA to pregnant dams or neonates (embryonal days 18 to postnatal days 14). In the NMDA-treated group, the significant increase in nuclei of apoptotic neuronal cells occurred in the dose-dependent manner in the lateral-ventral regions of the fetal cerebral cortex, reaching maximum values at 24 hours after treatment. On the other hand, the induction of apoptosis did not occur in the neonatal brain., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2003年07月, ［査読有り］

Hydroxyurea(HU)-induced apoptosis in the mouse fetal tissues
GH Woo; K Katayama; JY Jung; K Uetsuka; EJ Bak; H Nakayama; K Doi, Hydroxyurea (HU), a ribonucleotide reductase inhibitor, induces morphological anomalies in the central nervous system (CNS), craniofacial tissues and limb buds in animals, and neonatal respiratory distress in humans. In the present study, pregnant mice were treated with 400 mg/kg of HU at day 13 of gestation, and their fetuses were examined from I to 48 hours after treatment (HAT) to find a clue to clarify the mechanisms of HU-induced fetotoxicity and teratogenecity. At 6 and 12 HAT, a moderate to marked increase in the number of pyknotic cells was detected in the CNS and lung. A mild increase in the number of pyknotic cells was also found in the craniofacial mesenchymal tissues, limb buds and so on. These pyknotic cells had nuclei positively stained by the TUNEL method, which is widely used for the detection of apoptotic nuclei, and they also showed electron microscopic characteristics identical to those of apoptotic cells. The present results suggest that the HU-induced fetotoxicity is characterized by excess apoptotic cell death in the fetal tissues, and that such excess cell death in the fetal CNS, lung, craniofacial tissue and limb bud may have a certain relation to the later occurrence of morphological or functional anomalies reported in these tissues following HU-administration., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 2003年04月, ［査読有り］

Impaired liver regeneration after partial hepatectomy in db/db mice
H Yamauchi; K Uetsuka; T Okada; H Nakayama; K Doi, Fatty liver is the most common hepatic disorder in humans and supposed to be a cause of poor prognosis after liver transplantation and hepatic resection which could be resulted from impaired liver regeneration. This study was carried out to analyze the process of liver regeneration in db/db mice which show severe steatosis because of abnormal leptin receptor. We performed 70% partial hepatectomy (PH) on db/db mice and normal +m/+m mice, and then sacrificed the animals 1, 2, 3, 5, 7 and 10 days later. The liver samples were weighed and examined histologically or immunohistochemically. As a result, the liver mass restitution was significantly inhibited in db/db mice compared with +m/+m mice. The BrdU labelling index peaked at 2 days after PH in both strains, although the value was lower in db/db mice. After that, interestingly, it decreased to the control level at 5 days in +m/+m mice while the recovery was delayed in db/db mice. Similar sequence was also observed in the PCNA labelling index. In addition, the peak time of the mitosis index was 2 days and 5 days after PH in +m/+m mice and in db/db mice, respectively. Thus, although not significant, the proliferative response of hepatocytes to PH occurred somewhat more transient and sharply in +m/+m mice while it lasted somewhat longer in db/db mice. This suggests that db/db mice may be valuable as one of the animal models for the investigation of the effects of steatosis on the liver regeneration., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2003年03月, ［査読有り］

Effects of exogenous growth hormone (GH)-treatment on carbon tetrachloride (CCl₄)-induced acute liver injury in GH-deficient Mini rats.　　　　　　　　　　　　　　　
Okada T; Uetsuka K; Yamauch H; Tani Y; Endo K; Nakayama H; Doi K
Journal of Toxicologic Pathology., 2003年, ［査読有り］

DNA damages and changes in proliferating activity in the dorsal epidermis of UVB-irradiated WBN/ILA-Ht rats.　　　　　　　　　　　　　　　
Okada T; Uetsuka K; Nakayama H; Doi K
Journal of Toxicologic Pathology., 2003年, ［査読有り］

T-2 toxin-induced morphological changes in pregnant rats.　　　　　　　　　　　　　　　
Sehata S; Teranishi M; Atsumi F; Uetsuka K; Nakayama H; Doi K
Journal of Toxicologic Pathology., 2003年, ［査読有り］

1-beta-D-arabinofuranosylcytosine (Ara-C)-induced apoptosis in the rat fetal tissues and placenta.　　　　　　　　　　　　　　　
Yamauchi H; Katayama K; Yasoshima A; Uetsuka K; Nakayama H; Doi K
Journal of Toxicologic Pathology., 2003年, ［査読有り］

Dorsal skin responses to topical application with hydrogen peroxide in Mini and Wistar rats
A Umeda-Ikawa; Y Ishii; K Suzuki; K Uetsuka; H Nakayama; K Doi, Mini rats (Jcl: WistarTGN(ARGHGEN) 1 Nts) (MRs) are Wistar rat (WR)-derived transgenic rats in which the expression of growth hormone (GH) gene is suppressed under the presence of antisense RNA transgene. In order to evaluate the effects of GH-deficiency on the acute injury by external stimuli, the dorsal skin responses to a single topical application with 20% hydrogen peroxide (HPO), one of the environmental oxidative stressors, were histologically compared between male MRs and WRs of 8 weeks old, whose hair cycle was under the telogen phase. As a result, formation of granulation tissues, reepithelialization and regrowth of hair follicles were delayed in MRs compared with WRs. While hair follicles of MRs of this age are under a long-lasting telogen phase after their 2nd cycle, a new hair cycle started not only in the HPO-applied area but also in the solvent-applied area with a little time lag. These findings suggest that GH-deficiency may influence the skin responses to the external chemical stimuli., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2002年11月, ［査読有り］

Ethylnitrosourea induces apoptosis and growth arrest in the trophoblastic cells of rat placenta
K Katayama; M Ueno; H Takai; N Ejiri; K Uetsuka; H Nakayama; K Doi, Ethyl nitrosourea (ENU), a well known alkylating agent, induces congenital anomalies in fetuses when it is administered to pregnant animals. In previous studies, we reported that ENU induced apoptosis and growth arrest in fetal tissues and organs immediately after its administration to pregnant rats. In the present study, we investigated the histopathological changes of the placenta after ENU administration to pregnant rats on Day 13 of gestation (GD13) to obtain a clue for clarifying the role of the placenta in the process of fetal developmental disability induced by genotoxic stress. Apoptotic cells increased and DNA-replicating cells decreased in the trophoblastic cells in the placental labyrinth zone of the ENU-treated group by 3 h after treatment. The number of apoptotic cells peaked at 6 h after treatment and returned to control levels at 48 h after treatment. The number of DNA-replicating cells reached minimum levels at 6 h after treatment and returned to control levels at 48 h after treatment. By immunohistochemistry, p53-positive signals were observed in trophoblastic cells in the labyrinth zone of the ENU-treated group from 3 to 6 h after treatment. Significant decreases in fetal and placental weights were observed in the ENU-treated group at 2 days (GD15) and 8 days (GD21) after treatment. A reduction in the thickness of the labyrinth zone was histopathologically significant in the ENU-treated group. These results indicate that ENU induces apoptosis and growth arrest not only in fetal tissues, but also in trophoblastic cells in the rat placental labyrinth zone, and these placental changes may have roles in the induction of fetotoxicity and teratogenicity of ENU. Moreover, a possible involvement of p53 in the induction of apoptosis and growth arrest is suggested., SOC STUDY REPRODUCTION
BIOLOGY OF REPRODUCTION, 2002年08月, ［査読有り］

Cerebellar Myxoid type meningioma in a Shih Tzu dog
M Suzuki; H Nakayama; R Ohtsuka; A Yasoshima; K Katayama; K Uetsuka; M Ohta; S Matsunaga; H Ogawa; K Uchida; K Doi, A 6-year and 9 month-old, male, Shih Tzu dog showed ataxia and trembling. By MRI examination, a mass (I cm) was found in the right cerebellum. As the dog did not respond to radiation therapy, and showed a rise of intracranial pressure, he was euthanized. The cerebellar mass was soft and hemorrhagic. Histologically, the mass contained vimentin-positive spindle- or polyhedral-shaped cells arranged in a cord-like pattern. Mucinous materials were observed in the intercellular spaces. Ultrastructural examination revealed cell processes, microtubule-like structures and desmosomes. The case was diagnosed as myxoid type meningioma., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2002年02月, ［査読有り］

Granulocyte colony-stimulating factor exacerbates the acute lung injury and pulmonary fibrosis induced by intratracheal administration of bleomycin in rats
K Adachi; M Suzuki; T Sugimoto; S Suzuki; R Niki; A Oyama; K Uetsuka; H Nakamaya; K Doi, We investigated the effects of granulocyte colony-stimulating factor (G-CSF) on lung injury induced by intratracheal administration of bleomycin (BLM, 2 mg/200 mul) in rats. In experiment 1, G-CSF (10, 30 and 100 mug/kg/day, s.c.) was administered to rats treated with BLM or saline for 7 days starting immediately after BLM administration. In rats receiving G-CSF alone, a large number of neutrophils were noted in the pulmonary capillaries, although there were no lung lesions. In rats receiving BLM alone, diffuse alveolar damage was observed. The administration of G-CSF to BLM-treated rats increased the total lung lesion per unit of pulmonary parenchyma (total lung lesion %) along with increases in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion in a dose-dependent fashion. In experiment 2, 100 mug/kg/day of G-CSF was administered to rats treated with BLM or saline for up to 28 days starting immediately after BLM administration. The administration of 100 mug/kg/day of G-CSF to BLM-treated rats showed no effects at 14 days but it increased the lung lesion % and the score of lung fibrosis along with the increase in the number of neutrophils infiltrating in the pulmonary lesion at 28 days. These findings suggest that G-CSF administration to BLM-treated rats influenced and exacerbated the BLM-induced acute lung injury, and also exacerbated pulmonary fibrosis in a dose-dependent fashion. The exacerbation of lung injury coincided with the marked increase in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2002年02月, ［査読有り］

Effects of T-2 toxin treatment on CCl4-indueed hepatic necrosis in mice
Yukiko Muto; Yoshiro Tani; Taro Okada; Kei-Ichi Katayama; Michio Suzuki; Koji Uetsuka; Hiroyuki Nakayama; Kunio Doi, In the present study, effects of T-2 toxin, a kind of trichothecene mycotoxins produced by the genus Fusarium, on CCl4-induced hepatic necrosis and expression of CYP2E1, a major metabolizing enzyme for CCl4, were examined in the liver of mice. T-2 toxin treatment (4 mg/kg b.w.) at one day before CCl4 inoculation (1 ml/kg b.w.) suppressed hepatic necrosis almost completely, and T-2 toxin treatment at one day after CCl4 inoculation delayed the recovery of hepatic necrosis. Although not significant, T-2 toxin (4 mg/kg b.w.) lowered the total P450 contents until 48 hrs after treatment. In addition, Western blot analysis and immunohistochemical examinations revealed that T-2 toxin significantly suppressed CYP2E1 expression from 24 to 48 hrs after treatment while it did not influence CYP3A2 expression. These results suggest that pretreatment of T-2 toxin may suppress CCl4-induced hepatic necrosis through suppression of CYP2E1 expression., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 2002年, ［査読有り］

Liver responses to repeated doses of carbon tetrachloride in Mini and Wistar rats
Hiroko Shimizu; Koji Uetsuka; Taro Okada; Hiroyuki Nakayama; Kunio Doi, The liver responses to repeated doses of CC14 (0.5 mg/kg b.w., twice weekly) were compared between Wistar and Mini rats, in which the expression of growth hormone (GH) gene is suppressed by the presence of an antisense gene, at 2, 4, and 6 weeks after treatment (WAT). Fibrosis started earlier and its degree was severer in Mini rats than in Wistar rats, and hepatocyte damage was also severer in Mini rats than in Wistar rats. This corresponded well with the changes in serum AST and ALP levels. Oval cell proliferation in the fibrous septa of the liver of CC14treated Mini rats may also have some relations to such increase in ALP activity and fibrogenesis in Mini rats. The increase in the level of TGF-β1 mRNA was more prominent in Mini rats than in Wistar rats at 4 and 6 WAT, and this corresponded to the strain difference in the degree of liver fibrosis. The present results indicate that the responsibility of the liver to repeated doses of CCl4 was different between Mini rats and Wistar rats. Mini rats seem to be useful as a new tool in the investigation of regulatory mechanisms of GH on liver injury and regeneration., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 2002年, ［査読有り］

Development of apoptosis and changes in lymphocyte subsets in thymus, mesenteric lymph nodes and PEYER’s patches of mice orally inoculated with T-2 toxin.
Nagata T; Suzuki H; Ishigami N; Shinozuka J; Uetsuka K; Nakayama H; Doi K, Development of apoptosis and changes in lymphocyte subsets were examined mainly by flow cytometer in thymus, mesenteric lymph nodes and PEYER'S patches of mice up to 24 hours after oral inoculation with T-2 toxin (10 mg/kg). T-2 toxin attacked PEYER'S patches first, then mesenteric lymph nodes, and finally thymus in relation to the course of enteric absorption of orally inoculated T-2 toxin. The degree of lymphocyte apoptosis was prominent in the thymus, moderate in the PEYER'S patches, and somewhat mild in the mesenteric lymph nodes, suggesting the difference in lymphocyte population susceptible to T-2 toxin. As to the changes in lymphocyte subsets, CD4(+) CD8(+) T cells were most sensitive to T-2 toxin, and CD4(+) CD8(-) T cells were more severely depressed than CD4(-) CD8(+) T cells in the thymus. In the mesenteric lymph nodes, CD3(+) cells was more clearly affected than CD19(+) cells, and the numbers of CD4(+) and CD8(+) cells were similarly decreased. In the PEYER'S patches, the numbers of CD3(+), CD19(+), CD4(+) and CD8(+) cells were unexceptionally decreased. In addition, among IgM(+), IgG(+) and IgA(+) B cells, the number of IA(+) B cells which are more important in the mucosal immunity was most severely affected., URBAN & FISCHER VERLAG
Experimental and Toxicologic Pathology., 2001年09月, ［査読有り］

Carbon tetrachloride-induced acute liver injury in Mini and Wistar rats
H Shimizu; K Uetsuka; H Nakayama; K Doi, Acute liver injury induced by CCl4 injection (0.5 ml/kg b.w.) was compared between Mini and Wistar rats. Mini rats (Jcl:Wistar-TgN (ARGHGEN) 1 Nts strain) are Wistar-derived transgenic animals in which the expression of growth hormone (GH) gene is suppressed by the presence of an antisense transgene. The hepatic lesion appeared earlier and its recovery was delayed in Mini rats compared to in Wistar rats. The degree of the liver injury was more severe in Mini rats than in Wistar rats, and this corresponded well with the changes in serum AST level. Moreover, in accordance with the localization of CYP2E1-positive hepatocytes in the early stage after CCl4 treatment, the initial lesion characterized by ballooning of hepatocytes developed in the centrilobular zone in Wistar rats while it appeared in the middle zone in Mini rats. The changes in the percentage of PCNA-positive cells and the levels of HGF and TGF-beta1 mRNAs were clearly different between the two strains. These results indicate that the response of the liver to CCl4 is different between GH-suppressed Mini rats and Wistar rats., URBAN & FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 2001年04月, ［査読有り］

Early ultrastructural changes in the dorsal skin epidermis of wsistar-derived hypotrichotic WBN/ILA-£ft rats after UVB-irradiation
Valeria Malcotti; Akira Yasoshima; Koji Uetsuka; Hiroyuki Nakayama; Kunio Doi, Abstract: Early ultrastructural changes in the dorsal skin epidermis of Wistar-derived hypotrichotic WBN/ILA-Ht rats were examined for up to 12 hours after UVB-irradiation (10 kJ/m2) (12 HAI). The first change appeared at 3 HAI as swelling and/or vacuolation of mitochondria in the spinous and basal cells. The number of keratinocytes with such mitochondrial changes decreased thereafter. At 6 HAI, apoptosis of basal cells developed. These cells showed shrinkage of cell bodies with condensed or fragmented nuclei and dark cytoplasm due to densely packed bundles of tonofilaments. At the same time, keratinocytes with prominent nucleoli were also observed in the basal and suprabasal layers. At 12 HAL the number of keratinocytes with prominent nucleoli increased, and mild and focal intercellular edema developed mainly in the basal layer. In addition, a small number of apoptotic basal cells were still observed. The present ultrastructural findings supported the histopathological findings previously reported by our research group., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 2001年, ［査読有り］

Esophageal adenosquamous carcinoma in a cat
J Shinozuka; H Nakayama; M Suzuki; N Ejiri; K Uetsuka; M Mochizuki; R Nishimura; N Sasaki; K Doi, Esophageal carcinoma was observed in an eight-year-old, castrated male, Japanese domestic cat. Histologically, this neoplasm consisted of two different growth patterns, squamous cell carcinoma and adenocarcinoma. The results of immunohistochemical examination supported the fact that the two kinds of neoplastic cells have different characteristics. The tumor was, therefore, diagnosed as adenosquamous carcinoma. Esophageal tumors in the cat are very rare and, if any, neither adenocarcinoma nor adenosquamous carcinoma has been reported up to the present., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 2001年01月, ［査読有り］

Effects of granulocyte colony-stimulating factor on the development of inflammation in bleomycin-induced lung injury
Kenji Adachi; Masami Suzuki; Tetsurou Sugimoto; Koji Uetsuka; Hiroyuki Nakayama; Kunio Doi, We investigated the effects of granulocyte colony-stimulating factor (G-CSF) on the development of inflammation in the lung induced by intratracheal administration of bleomycin (BLM, 2 mg/200 μl) in rats. G-CSF (100 μg/kg/day) was subcutaneously administered for 7 days starting from 3 (inflammatory phase) or 14 days (fibrogenic phase) after BLM administration. The administration of G-CSF to BLM-treated rats in the inflammatory phase increased the total lung lesion per unit pulmonary parenchyma, the score of lung fibrosis, the peripheral neutrophil count and the number of neutrophils infiltrating the pulmonary lesion, as well as the severity of other histopathologic changes (edema, hemorrhage and proliferation of myofibroblasts in the alveolar lumen, hyperplasia of bronchial and alveolar epithelial cells, and bronchiolization). Although increases in the peripheral neutrophil count and the number of neutrophils infiltrating the pulmonary lesion were observed following treatment with G-CSF, there were no further increases on lung injury in the fibrogenic phase in BLM-treated rats. These findings suggest that the effects of neutrophils on the lung in the rats treated with G-CSF may be closely related not only to the number of neutrophils, but also to the inflammatory phase of BLM-induced pulmonary lesions., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 2001年, ［査読有り］

Picryl chloride-induced contact dermatitis in IQI/Jic male mice.
Jung J-Y; Saegusa J; Uetsuka K; Nakayama H; Doi K, Ear skin responses to picryl chloride (PCL) were examined in IQI/Jic male mice. Following the sensitization with PCL to the shaved abdominal skin, the left ear was topically applied with PCL at 4, 11, 18 and 25 days later. The ear swelling responses tended to shift from delayed type to immediate one after the 4th application. The peak time of histopathological changes characterized by edema and inflammatory cell infiltration corresponded well with that of ear swelling responses, and marked elevation of total serum IgE levels and prominent increase in number of mast cells were observed after the 4th application. The numbers of CD4⁺ cells and Mac-1⁺ cells began to increase after the 1st application and much more increased after the 4th application while CD8⁺ cells increased after the 4th application. MHC class II⁺ cells increased mainly in the dermis after the 4th application, but CD19 ⁺ cells, i.e. B lymphocytes, were negligible throughout the experimental period. The present results suggest that not only CD4⁺ cells but also CD8⁺ cells may participate in the induction of PCL-induced contact dermatitis in IQI/Jic male mice and that the shift from Th1-dominant response to Th2-dominant one may occur following the repeated application., 日本毒性病理学会
Journal of Toxicologic Pathology., 2001年, ［査読有り］

Apoptotic cell death and cell proliferative activity in the rat fetal central nervous system from dams administered with ethylnitrosourea (ENU)
K Katayama; K Uetsuka; N Ishigami; H Nakayama; K Doi, Ethylnitrosourea (ENU), a well known DNA alkylating agent, induces anomalies in the central nervous system (CNS), craniofacial tissues and male reproductive organs, and the enhancement of apoptosis is found in these tissues immediately after the administration of ENU (Katayama et al., 2000a). In this study, pregnant rats were treated with 60mg/kg of ENU at day 13 of gestation, and kinetics of apoptotic cells, mitotic cells and bromodeoxyuridine (BrdU)-positive cells in the fetal CNS were examined from 3 to 48 hours after the treatment (HAT). From 3 HAT, a significant increase in the number of apoptotic cells and a significant decrease in the number of mitotic cells were detected in the fetal CNS, and BrdU-positive cells significantly decreased in accordance with the increase in the number of apoptotic cells. The present results strongly suggest that both excess cell death by apoptosis and cell growth arrest indicated by decreased number of mitotic cells and BrdU-positive cells may have a close relation to the later occurrence of microencephaly following ENU-administration, and that ENU affects mainly S-phase cells and causes apoptosis., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 2001年01月, ［査読有り］

Development of apoptosis and changes in apoptosis-related genes expression in the thymus of nivalenol-treated mice
Amnart Poapolathep; Kazuhiko Suzuki; Kei-Ichi Katayama; Ryoichi Ohtsuka; Takayuki Nagata; Koji Uetsuka; Hiroyuki Nakayama; Kunio Doi, Nivalenol (NIV) is a potent toxic trichothecene mycotoxin produced by Fusarium nivale, and our previous study clarified first that NIV induces apoptosis in lymphoid organs of mice in a dose-dependent manner. In this study, the development of apoptosis and changes in the expression of apoptosis-related genes (fas, c-fos, c-jun, p53, bcl-2 and c-myc) mRNAs were examined in the thymus of mice for up to 12 hours after inoculation (HAI) of NIV (15 mg/kg b.w.) to elucidate the relationship between the molecular genetic regulatory mechanisms and NIV-induced lymphocyte apoptosis. The number of apoptotic lymphocytes evaluated by TUNEL method clearly increased from 3 HAI and peaked at 9 HAL The DNA ladder formation by agarose gel electrophoresis was clearly observed at 6 and 9 HAL The expression of c-fos and c-jun mRNAs clearly elevated from 0.5 HAI and peaked at 1 HAL The expression of fas, p53, bcl-2 and c-myc mRNAs showed no significant changes. These results indicate that c-fos and c-jun may play an important role in NIV- induced lymphocyte apoptosis., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 2001年, ［査読有り］

Ethylnitrosourea (ENU)-induced apoptosis in the rat fetal tissues
K Katayama; N Ishigami; K Uetsuka; H Nakayama; K Doi, Ethylnitrosourea (ENU), a well known DNA alkylating agent, induces anomalies in the central nervous system (CNS), craniofacial tissues and male reproductive organs. In this study, pregnant rats were treated with 60 mg/kg ENU at day 13 of gestation, and their fetuses were examined from 1 to 48 hours after treatment (HAT) to find a clue for clarifying the mechanisms of the ENU fetotoxicity and teratogenicity. From 3 to 12 HAT, the moderate to marked increase in the number of pyknotic cells was detected in the fetal CNS, craniofacial mesenchymal tissues, gonads and so on. These pyknotic cells had nuclei positively stained by the TUNEL method, which is widely used for the detection of apoptotic nuclei, and they also showed electron microscopic characteristics identical to those of apoptotic cells. The present results strongly suggest that excess cell death by apoptosis in the fetal CNS, craniofacial tissues and gonads may have a close relation to the later occurrence of anomalies reported in these tissues following ENU- administration., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 2000年07月, ［査読有り］

Cellular proliferative and telomerase activity in canine mammary gland tumors
Y Funakoshi; H Nakayama; K Uetsuka; R Nishimura; N Sasaki; K Doi, In canine mammary tumors, we examined the telomerase activity, proliferative activity by proliferative cell nuclear antigen (PCNA) immunohistochemistry, and percentage of apoptotic cells by the deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method. The relationship between these measures and histopathologic malignancy was also investigated. PCNA index was highest in malignant tumors (adenocarcinoma: 27.0%; malignant mixed tumor: 15.7%), followed by benign tumors (adenoma: 4.4%; benign mixed tumor: 5.3%), hyperplasia (2.1%), and normal mammary gland (0.9%). In adenoma and adenocarcinoma, papillary and solid types showing higher cellularity tended to have higher PCNA indices than did cystic and tubular types. Although the TUNEL index was <1% in all cases, the relationship between this measure and histopathologic diagnosis showed the same tendency as observed in PCNA immunostaining. Telomerase activity was detectable in all adenomas, benign mixed tumors, and adenocarcinomas examined. In contrast, all normal mammary glands, hyperplasias, and malignant mixed tumors were negative for telomerase. Relative telomerase activity (RTA) of adenocarcinoma (56.5) was significantly higher than that of adenoma (27.8) and benign mixed tumor (33.9), and a significant positive correlation (P < 0.001) was noted between RTA and PCNA index. No significant correlations were noted between either PCNA or TUNEL index and clinical features such as metastasis and tumor diameter. PCNA index and telomerase activity may be useful markers for judging malignancy of canine mammary tumors., AMER COLL VET PATHOLOGIST
VETERINARY PATHOLOGY, 2000年03月, ［査読有り］

Age-related changes in susceptibility of rat brain slice cultures including hippocampus to encephalomyocarditis virus
WP Su; A Ueno-Yamanouchi; K Uetsuka; H Nakayama; K Doi, Replication of the D variant of encephalomyocarditis virus (EMC-D) and its cytopathic effects were studied in the brain slice cultures including hippocampus (hippocampal slice) obtained from postnatal 1-, 4-, 7-, 14-, 28- and 56-day-old Fischer 344 rats. At 0, 12, 24, 36 and 48 h after infection, virus titres of the slices and culture media were assayed. Viral replication was observed in cultures from 1-to 28-day-old rats, and the highest titre was recorded in the slice and culture medium from the youngest rat. The peak of virus titre decreased with age and no distinct viral replication was observed in the cultures from 56-day-old rats. Light microscopy revealed that degenerative and necrotic changes appeared in the infected hippocampal slices from 1- to 28-day-old rats, and the changes became less prominent with age. In situ hybridization and indirect immunofluorescence staining showed that positive signals of viral RNA and antigen were prominent in younger rats and decreased with age. These results suggest that an age-related decrease in the susceptibility of rat brain to EMC-D is less related to the maturation of the immune system but possibly to that of the neurone., BLACKWELL SCIENCE LTD
INTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, 1999年12月, ［査読有り］

Apoptosis in the developing mouse embryos from T-2 toxin-inoculated dams
N Ishigami; J Shinozuka; K Katayama; K Uetsuka; H Nakayama; K Doi, T-2 toxin (3mg/kg b.w.) was orally inoculated to pregnant mice at 11 days of gestation to examine the effect of T-2 toxin on the developing embryos. At 24 hours after T-2 toxin-inoculation, moderate pyknosis or karyorrhexis was generally observed in some layers of the central nervous system, caudal sclerotomic segment, caudal region of the tongue to pharyngeal- to laryngeal-mesenchyma, trachea and facial mesenchyma. These pyknotic or karyorrhectic nuclei were strongly stained by the modified TUNEL method widely used for the in situ detection of apoptotic nuclei and also showed ultrastructural changes characteristic for apoptosis. This is the first report of mycotoxin-induced apoptosis in embryos., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 1999年07月, ［査読有り］

Kinetics of transforming growth factor-beta 1 and extracellular matrix in renal tubulointerstitial lesions of mercuric chloride-treated Brown Norway rats
K Suzuki; K Uetsuka; H Nakayama; K Doi, Renal tubulointerstitial lesions in mercuric chloride(HgCl2)-treated Brown Norway rats were investigated focusing on the kinetics of transforming growth factor-beta 1 (TGF-beta 1) and extracellular matrix (ECM). Rats were injected with 1 mg/kg b.w. of HgCl2 at days 0, 2, and 4, and 5 rats were killed at days 2, 4, 6, 8, 10, and 20, respectively. TGF-beta 1 mRNA expression in the renal cortex measured by competitive RT-PCR method reached a peak at day 6, mildly decreased at days 8 and 10, and increased again toward day 20. Signals of TGF-beta 1 mRNA examined by in situ hybridization method were recognized in the regenerative tubular epithelium at day 6, and in both tubular epithelium and infiltrated mononuclear cells at day 20. After tubular injury, strong immunoreactivity to TGF-beta 1 protein was found in desquamated tubular epithelial cells. Then, positive staining was found in the regenerative tubular epithelial cells. Later, infiltrated mononuclear cells also became positive for TGF-beta 1 protein. In the ECM, deposition of fibronectin was prominent throughout the experimental period. In conclusion, this strongly suggests that TGF-beta 1 derived from tubular epithelial cells and some macrophages might be related to the development of renal interstitial fibrosis in HgCl2-treated BN rats., BLACKWELL SCIENCE LTD
INTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, 1999年06月, ［査読有り］

Generalized dermal dysplasia and perifollicular mucinosis in a dog
K Uetsuka; M Suzuki; M Suehiro; N Ejiri; R Nishimura; N Sasaki; H Nakayama; K Doi, Multiple nodules 0.5-2 cm in diameter were observed in the dorsal skin of a 1.5-year-old male castrated mongrel dog. Histopathologically, perifollicular mucinosis and clusters of small cells with a dense, round nucleus and scant cytoplasm were seen. The small cell clusters were present in the dermis near the epidermal basal layer or surrounding hair follicles. These cells demonstrated different staining patterns for vimentin and keratin depending on their location; ultrastructurally, they were poor in organelles and connected to each other with cytoplasmic projections. These findings suggested that they were immature epithelial cells. The case was diagnosed as generalized dermal dysplasia and perifollicular mucinosis., AMER COLL VET PATHOLOGIST
VETERINARY PATHOLOGY, 1999年01月, ［査読有り］

Renal tubulointerstitial lesions in mercuric chloride-treated Brown Norway rats.　　　　　　　　　　　　　　　
Suzuki K; Uetsuka K; Nakayama H; Doi C; Doi K
Journal of Toxicologic Pathology., 1999年, ［査読有り］

Glycoconjugate expression in follicle-associated epithelium (FAE) covering the nasal-associated lymphoid tissue (NALT) in specific pathogen-free and conventional rats
KI Jeong; K Uetsuka; H Nakayama; K Doi, We examined lectin-histochemically the glycoconjugate expression in the follicle-associated epithelium (FAE) covering the nasal-associated lymphoid tissue (NALT) in the rat under specific pathogen-free (SPF) and conventional (CV) conditions and compared the results for SPF and CV rats as well as for membranous (M) cells and adjacent ciliated respiratory epithelial (CRE) cells in FAE. N-acetylgalactosamine-specific lectins, Dolichos biflorus (DBA), Helix pomatia (HPA), Glycine max (SBA) and Vicia villosa (VVA), and alpha-L-fucose-specific lectin, Ulex europaeus (UEA-I), preferentially bound to M cells mainly in the luminal surface compared with CRE cells in SPF rats, whereas DBA and UEA-I showed signs of preferential binding to the apical and basolateral cytoplasm as well as to the luminal surface of M cells in CV rats. In addition, HPA, SEA and VVA more frequently and extensively labeled M cells than CRE cells in CV rats with the same subcellular staining pattern as DBA and UEA-I. On the whole, the changes in lectin binding frequency and strength were more prominent in M cells than in CRE cells in both SPF and CV rats. The present results indicate that DBA and UEA-I are useful as markers of M cells in NALT. Furthermore, the pattern of expression of carbohydrate residues recognized by such lectins in SPF and CV rats suggests that M cells are highly sensitive to environmental changes., INT PRESS EDITING CENTRE INC
EXPERIMENTAL ANIMALS, 1999年01月, ［査読有り］

Cerebral amyloid angiopathy in an aged great spotted woodpecker (Picoides major)
H Nakayama; KI Katayama; A Ikawa; K Miyawaki; J Shinozuka; K Uetsuka; SI Nakamura; N Kimura; Y Yoshikawa; K Doi, A male great spotted woodpecker (Picoides major), which was at least 16 years old, died due to general weakening. Cerebral vascular walls, including capillaries, were positively stained with Congo red with green-gold birefringence, and some of which showed a severe deposition of the Congophilic materials resulting in a corona-like fibrillar radiating structure. The Congophilic materials were positive for beta amyloid protein, but negative for prion protein. Only a few senile plaque-like structures were observed in the cortex by PAM stain and beta amyloid immunostain. The present case is the first observation of cerebral amyloid angiopathy in avian species and will indicate the presence of such age-related cerebral lesions also in birds. (C) 1999 Elsevier Science Inc. All rights reserved., ELSEVIER SCIENCE INC
NEUROBIOLOGY OF AGING, 1999年01月, ［査読有り］

Cutting edge: BASH, a novel signaling molecule preferentially expressed in B cells of the bursa of Fabricius
R Goitsuka; Y Fujimura; H Mamada; A Umeda; T Morimura; K Uetsuka; K Doi; S Tsuji; D Kitamura, The bursa of Fabricius is a gut-associated lymphoid organ that is essential for the generation of a diversified B cell repertoire in the chicken, We describe here a: novel gene preferentially expressed in bursal R cells. The gene encodes an 85-kDa protein, designated-BASH (B cell adaptor containing SH2 domain), that contains N-terminal acidic domains with SH2 domain-binding phosphotyrosine-based moths, a proline-rich domain, and a C-terminal SH2 domain, BASH shows a substantial sequence similarity to SLP-76, an adaptor protein functioning in TCR-signal transduction, BASH becomes tyrosin-phosphorylated with the B cell Ag receptor (BCR) cross-link or by coexpression with Syk and Lyrr and associates with signaling molecules including Syk and a putative chicken Shc homologue, Overexpression of BASH results in suppression of the NF-AT activation induced by BCR-cross-linking. These findings suggest that BASH is involved in BCR-mediated signal transduction and could play a critical role in B cell development in the bursa., AMER ASSOC IMMUNOLOGISTS
JOURNAL OF IMMUNOLOGY, 1998年12月, ［査読有り］

A rare case of patent ductus arteriosus in a dog with glomerular mesangiolysis
K Katayama; J Oh; K Uetsuka; R Nishimura; H Nakayama; N Sasaki; K Doi, Rare case of patent ductus arteriosus (PDA) was observed in a 2-year and 9 month-old Miniature Dachshund which had been diagnosed as severe cardiac failure. Pulmonary artery from the right ventricle connected to dilated ductus arteriosus, and pulmonary artery-ductus arteriosus-descending aorta formed a continuous duct, which seemed to be the main route of bloodstream. Ascending aorta from the left ventricle was hypoplastic, and connected to the dilated ductus arteriosus. Glomerular mesangiolysis due to heart failure was also observed in the kidney., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1998年11月, ［査読有り］

T-2 toxin-induced apoptosis in hematopoietic tissues of mice
J Shinozuka; M Suzuki; N Noguchi; T Sugimoto; K Uetsuka; H Nakayama; K Doi, We examined T-2 toxin-induced lesions in the bone marrow and splenic red pulp as many as 48 hr after oral inoculation with 10 mg/kg body weight of T-2 toxin in female ICR:CD-1 mice. Histopathologically, the bone marrow and splenic red pulp showed a significant hypocellularity. In the bone marrow, the number of myelocytes significantly decreased due to the loss of immature granulocytes, erythroblasts, and lymphocytes. The nuclei of the remaining cells showing pyknosis or karyorrhexis were positively stained by the TdT-mediated dUTP nick end labeling (TUNEL) method, and these TUNEL-positive cells showed ultrastructural characteristics of apoptosis. With agarose gel electrophoresis, DNA ladders were clearly detected in bone marrow samples. The number of TUNEL-positive cells in splenic red pulp increased earlier than it did in the splenic white pulp. Thus, T-2 toxin induced-lesions in the hematopoietic tissues and in the lymphoid tissues were brought about by apoptosis of component cells. We believe that damage to the hematopoietic microenvironment may also play an indirect role in the induction of apoptosis in the bone marrow., SOC TOXICOLOGIC PATHOLOGISTS
TOXICOLOGIC PATHOLOGY, 1998年09月, ［査読有り］

Lectin histochemistry of dorsal skin of Wistar-derived hypotrichotic WBN/Ila-Ht rats
S Iwamoto; C Doi; K Uetsuka; H Nakayama; K Doi, A lectin histochemical study was carried out on the dorsal skin of Wistar-derived hypotrichotic WBN/Ila-Ht rats (HtRs) and Wistar rats (WRs) at 3, 7 and 24 weeks of age to clarify the lectinhistochemical characteristics of the skin during their development. The lectins examined were Concanavalia ensiformis (Con A), Dolichos biflorus agglutinin (DBA), Griffonia simpliciolia (GS-I), Helix pomatia agglutinin (HPA), Arachis hypogaea (PNA), Glycine maximus agglutinin (SBA), Ulex europeus agglutinin (UEA-I) and Triticum vulgaris agglutinin (WGA). None of the nucleated cell layers of the epidermis had DBA-binding sites, but they were all stained intensely with HPA and weakly with con A irrespective of the strain and age of the rats. As to the other 5 lectins, the intensity of binding activity was generally weaker in HtRs than in WRs and at 3 weeks of age than at 7 or 24 weeks of age, respectively. Among them, UEA-I mainly bound to the spinous cell layer but not to the basal cell layer, suggesting that alpha-L-fucose would be expressed on the cell surface according to the differentiation of keratinocytes. In addition, GS-I, HPA and UEA-I bound to the hair follicle epithelium and many lectins stained sebaceous gland epithelial cells. In conclusion, except for the binding intensity of some lectins, there were no specific differences between HtRs and WRs in the lectinhistochemical characteristics of the dorsal skin epidermis. The present data on the rat skin would be useful from the viewpoint of comparative lectinhistochemistry., INT PRESS EDITING CENTRE INC
EXPERIMENTAL ANIMALS, 1998年07月, ［査読有り］

Changes in location and number of tartrate-resistant acid phosphatase (TRAP)-positive cells during the development of type II collagen-induced arthritis in DBA/1J mice
M Suzuki; M Suzuki; K Uetsuka; J Shinozuka; H Nakayama; K Doi, The authors investigated changes in the location and number of osteoclasts and their precursors during the development of articular lesions in type ii collagen-induced arthritis in mice using tartrate-resistant acid phosphatase (TRAP) staining. The limb joints were examined at 6 to 15 weeks after the second immunization. The number of TRAP-positive cells increased as the articular lesions progressed. TRAP-positive macrophagelike cells were found in the hyperplastic synovial tissue and bone marrow stroma in the early stage. In the advanced stage, in addition to many TRAP-positive osteoclasts on the bone surface, TRAP-positive macrophage-like cells were observed in the pannus apart from the bone surface in the pannus-joint junctions. The above mentioned TRAP-positive macrophage-like cells are considered to be osteoclast precursors., INT PRESS EDITING CENTRE INC
EXPERIMENTAL ANIMALS, 1998年07月, ［査読有り］

Eight-year observation and comparative study of specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) subtypes A and B: Terminal acquired immunodeficiency syndrome in a cat infected with FIV Petaluma strain
M Kohmoto; K Uetsuka; Y Ikeda; Y Inoshima; M Shimojima; E Sato; G Inada; T Toyosaki; T Miyazawa; KN Doi; T Mikami, Three specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) strains Petaluma, TM1 and TM2, respectively were observed for over 8 years. Without showing any significant clinical signs of immunodeficiency syndrome (AIDS) For 8 years and 4 months of asymptomatic phase, the Petaluma-infected cat exhibited severe stomatitis/ging/vitis, anorexia, emaciation, hematological and immunological disorders such as severe anemia, lymphopenia, thrombocytopenia, and decrease of CD4/CD8 ratio to 0.075, and finally died with hemoperitoneum at 8 years and 8 months post-infection. Histopathological studies revealed that the cat had systemic lymphoid atrophy and bone marrow disorders indicating acute myelocytic leukemia (aleukemic type). Plasma viral titer of the cat at AIDS phase was considerably high and anti-FIV antibody titer was slightly low as compared with the other FIV-infected cats. In addition, immunoblotting analysis using serially collected serum/plasma samples of these cats revealed that antibodies against FIV proteins were induced in all the infected cats, however in the Petaluma-infected cat anti-Gag antibodies disappeared during the asymptomatic period. These results suggested that plasma viral load and anti-FIV Gag antibody response correlated with disease progression, and supported FIV-infected cats as a suitable animal model of human AIDS., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1998年03月, ［査読有り］

Strain.
KOHMOTO Mariko; UETSUKA Koji; IKEDA Yasuhiro; INOSHIMA Yasuo; SHIMOJIMA Masayuki; SATO Eiji; INADA Gohki; TOYOSAKI Tomoko; MIYAZAWA Takayuki; DOI Kunio; MIKAMI Takeshi, Three specific pathogen-free cats experimentally infected with feline immunodeficiency virus (FIV) strains Petaluma, TM1 and TM2, respectively were observed for over 8 years. Without showing any significant clinical signs of immunodeficiency syndrome (AIDS) for 8 years and 4 months of asymptomatic phase, the Petaluma-infected cat exhibited severe stomatitis/gingivitis, anorexia, emaciation, hematological and immunological disorders such as severe anemia, lymphopenia, thrombocytopenia, and decrease of CD4/CD8 ratio to 0.075, and finally died with hemoperitoneum at 8 years and 8 months post-infection. Histopathological studies revealed that the cat had systemic lymphoid atrophy and bone marrow disorders indicating acute myelocytic leukemia (aleukemic type). Plasma viral titer of the cat at AIDS phase was considerably high and anti-FIV antibody titer was slightly low as compared with the other FIV-infected cats. In addition, immunoblotting analysis using serially collected serum/plasma samples of these cats revealed that antibodies against FIV proteins were induced in all the infected cats, however in the Petaluma-infected cat anti-Gag antibodies disappeared during the asymptomatic period. These results suggested that plasma viral load and anti-FIV Gag antibody response correlated with disease progression, and supported FIV-infected cats as a suitable animal model of human AIDS., JAPANESE SOCIETY OF VETERINARY SCIENCE
Journal of Veterinary Medical Science, 1998年

Cats are protected against feline immunodeficiency virus infection following vaccination with a homologous AP-1 binding site-deleted mutant
M Kohmoto; T Miyazawa; E Sato; K Uetsuka; Y Nishimura; Y Ikeda; G Inada; K Doi; T Mikami, Following establishment, via the vaginal route, of infection with an AP-1 binding-site deleted mutant (Delta AP-1) of feline immunodeficiency virus (FIV), cats were challenged with a homologous intact strain (TM2) of FIV. The cats were observed for 23 weeks to evaluate the efficacy of the Delta AP-1 against the homologous TM2 strain challenge. These two viruses were differentiated by Southern blotting after amplification of proviral DNA by semi-nested polymerase chain reaction in DNAs of peripheral blood mononuclear cells and tissues. A TM2-specific band was detected in one cat exposed to but not infected with Delta AP-1, but not in two Delta AP-1-infected. These results indicate that Delta AP-1 could protect against subsequent challenge with homologous FIV TM2 strain., SPRINGER-VERLAG WIEN
ARCHIVES OF VIROLOGY, 1998年, ［査読有り］

Immunohistochemical study on hepatic component cells and extracellular matrix in Wistar rats with galactosamine-induced acute hepatitis
Michio Suzuki; Koji Uetsuka; Junko Shinozuka; Hiroyuki Nakayama; Kunio Doi, After a single administration of D-galactosamine hydrochloride (GaIN) in Wistar rats, hepatocyte apoptosis was followed by proliferation of oval cells, resulting in formation of small bile duct-like structures. An activation of Ito cells seems to participate in the formation of small bile duct-like structures through the production of laminin and fibronectin which are primary components of basement membrane., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 1998年, ［査読有り］

Rapid induction of liver cirrhosis in mini rats by thioacetamide
Yoshiro Tani; Shinji Murata; Sachiko Kajiwara; Hatsue Mizuno; Junichiro Fukushige; Tsuneraichi Hosokawa; Koji Uetsuka; Hiroyuki Nakayama; Kunio Doi, A histopathological study was carried out on the liver of thioacetamide (TAA)-treated rats of Jcl: Wistar-TgN(ARGHGEN)1Nts strain (Mini rats), in which the expression of growth hormone (GH) gene is suppressed by the presence of antisense transgene, resulting in the reduction of plasma GH levels. Male Wistar (the parental strain of Mini rats) and Mini rats of 4-6 weeks of age were treated with 0.03% TAA in drinking water for up to 8 weeks. The survival rates of Wistar and Mini rats were 91% and 89%, respectively. Plasma alkaline phosphatase activity and total bilirubin concentration in TAA-treated Mini rats increased to a greater extent than those in TAA-treated Wistar rats. Histopathologically, the liver of TAA-treated Mini rats revealed proliferation of 'atypical' hepatocytes throughout the course of the experiment. In addition, fibrosis from the periportal area to the centrilobular region associated with oval cell proliferation was observed at 2 weeks. At 8 weeks, cirrhosis accompanied with proliferation of bile duct epithelial cell-like cells was evident. In the liver of Wistar rats, there were only atypical hepatocytes and slight fibrosis without cirrhotic features even at 8 weeks. These results demonstrate a great advantage in the use of Mini rats to produce liver cirrhosis within a short term and suggest some possible relationships between GH deficiency and experimental cirrhogenesis., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 1998年, ［査読有り］

5-azacytidine-induced apoptosis in the central nervous system of developing rat fetuses.　　　　　　　　　　　　　　　
Lu D-P; Nakayama H; Shinozuka J; Uetsuka K; Taki R; Doi K
Journal of Toxicologic Pathology., 1998年, ［査読有り］

Lectin histochemistry of normal and neoplastic canine mammary gland epithelial cells
Noriko Ejiri; Koji Uetsuka; Etsuko Nishida; Hiroyuki Nakayama; Kunio Doi, Lectin histochemical study was done on canine mammary gland epithelial cells of 6 groups (normal, hyperplasia, adenoma, adenocarcinoma, benign mixed tumor and malignant mixed tumor). Among ten lectins used, three lectins showed an apparent alteration in the binding activity following neoplastic changes. Namely, Dolichos biflorus and Helix pomatia specific for terminal N -acetylgalactosamine residue bound more strongly to the surface of epithelial cells in adenocarcinoma and malignant mixed tumor. In addition, Ulex europaeus specific for terminal fucose residue showed stronger binding activity in mammary neoplasma irrespective of malignancy., Japanese Society of Toxicologic Pathology
Journal of Toxicologic Pathology, 1998年, ［査読有り］

T-2 toxin-induced apoptosis in intestinal crypt epithelial cells of mice
GM Li; J Shinozuka; K Uetsuka; H Nakayama; K Doi, The characteristics of T-2 toxin-induced cell damage in the intestinal crypt epithelia was investigated in mice. Following T-2 toxin-inoculation (0, 2.5, 5 and 10 mg/kg b. w.), dead cells showing pyknosis were sporadically observed in the crypt epithelia, and the nuclei of these cells were strongly stained by the modified TUNEL method which detects fragmented DNA in situ. Electron microscopically, the dead cells were characterized by shrinkage of the cell body and condensation of nuclear chromatin frequently along the nuclear membrane, and such nuclei were sometimes fragmented into small pieces. These morphological characteristics are well consistent with those of apoptosis. The mitotic index in the crypt epithelia drastically decreased at 6 hours after T-2 toxin-inoculation (6 HAI), but thereafter it recovered to almost the same value with that in control mice at 48 HAI. On the other hand, the apoptotic index in the crypt epithelia increased with the lapse of time. Clear mouse strain-and sex-differences were detected in the apoptotic index but not in the mitotic index. This is the first report that T-2 toxin caused apoptotic cell death in the intestinal crypt epithelial cells., GUSTAV FISCHER VERLAG
EXPERIMENTAL AND TOXICOLOGIC PATHOLOGY, 1997年12月, ［査読有り］

Prolonged oval cell proliferation with Ito cell activation and extracellular matrix accumulation in galactosamine-induced acute hepatitis in mini rats
K Uetsuka; M Suzuki; H Nakayama; K Doi, Histopathological and immunohistochemical studies were carried out on D-galactosamine (GalNAc)induced acute hepatitis in rats of the JCl: Wistar TgN (ARGHGEN) 1 Nts strain (Mini rats), in which expression of the growth hormone gene is suppressed by an antisense transgene. Hepatitis characterized by hepatocellular acidophilic necrosis with inflammatory cell infiltration was most prominent at 2 days after GalNAc (100.0mg/kg)-injection, when proliferation of Ito cells and deposition of fibronectin and laminin were found along the sinusoidal linings. At 72 hours after GalNAc-injection, Ito cell proliferation with deposition of laminin and fibronectin became more prominent, and marked proliferation of small epithelial cells was observed in the periportal area. At 7 days after GalNAc-injection, quite a number of alpha-smooth muscle actin-positive Ito cells, surrounded by abundant fibronectin, laminin and type IV collagen, were still observed in close juxtaposition to rapidly proliferating small epithelial cells. The small epithelial cells were found to be positive for both alpha-fetoprotein and cytokeratin 7 and were therefore considered to be so-called oval cells. The results suggest that there may be some relation between oval cell proliferation, Ito cell activation and extracellular matrix accumulation in GalNAc-induced acute hepatitis in Mini rats., F HERNANDEZ
HISTOLOGY AND HISTOPATHOLOGY, 1997年10月, ［査読有り］

Immunohistochemical study on type II collagen-induced arthritis in DBA/1J mice
M Suzuki; K Uetsuka; M Suzuki; J Shinozuka; H Nakayama; K Doi, We performed immunohistochemical examinations on type II collagen-induced arthritis (CIA) mice, focusing attention on the changes in distribution of plasma proteins and extracellular matrix materials (ECM) and in expression of adhesion molecules. The limb joints of male DBA/1J mice immunized with bovine type II collagen were obtained at 6 to 20 weeks after the first immunization. In the early stage of CIA, deposition of fibrin, IgG, von Willebrand factor (vWF) and fibronectin was detected on the surface of the synovial lining layer and articular cartilage and in the articular cavity. In the stage of pannus formation, prominent proliferation of ICAM-1-positive capillaries and marked infiltration of LFA-1-positive neutrophils were observed in the pannus. The superficial portion of the pannus and basement membranes of proliferated capillaries were strongly positive for type IV collagen and laminin. In the late stage, the pannus invaded and destroyed articular cartilage and subchondral bone, and strongly positive immunostainabilities for both lysozyme and fibronectin were observed on the surface of the pannus and at the junctional portion between the pannus and the cartilage. The present immunohistochemical findings on the distribution of plasma proteins and ECM materials and the expression of adhesion molecules in CIA mice were similar to those in rheumatoid arthritis (RAJ in many aspects. This suggests that CIA is a useful model for the investigation of RA., JAPAN EXPER ANIMAL RES ASSN
EXPERIMENTAL ANIMALS, 1997年10月, ［査読有り］

Amyloid deposits in the gastrointestinal tract of aging dogs
Y Tani; K Uchida; K Uetsuka; S Nakamura; H Nakayama; N Goto; K Doi, Pathologic features, immunohistochemical characteristics, and incidence of gastrointestinal amyloid deposition were investigated in 78 canine necropsies, including 43 males and 35 females ranging from 2 months to 20 years of age. In 31 of 78 dogs, amyloid protein was accumulated in or around vessel walls located in submucosal and muscular layers of the alimentary tract and mesenteric tissues. These deposits did not seem to affect the function(s) of the digestive tract histologically. The deposits showed congophilia and green birefringence under polarized microscopy. Transmission electron microscopy revealed randomly arranged amyloid fibrils 6-10 nm in diameter around a blood vessel. Congophilia was retained even after oxidation with potassium permanganate, suggesting that this protein is not the AA form of amyloid. Although immunohistochemical studies were performed using antibodies to canine amyloid apolipoprotein AI and synthetic human beta amyloid peptide, the immunochemical nature of the protein was not determined. The mean ages of both male and female dogs with gastrointestinal amyloid deposition were higher than those of their normal counterparts (P < 0.001), and the incidence of deposition in elderly dogs was higher than that in young dogs (P < 0.005). However, there was no sex predilection with respect to both the mean age of the dogs with deposition and the incidence of occurrence. The mechanism(s) and importance of this form of amyloid deposition remain unclear., SAGE PUBLICATIONS INC
VETERINARY PATHOLOGY, 1997年09月, ［査読有り］

Protective effects of a novel quinone derivative, E3330, on mouse hepatitis virus (MHV)-induced chronic hepatitis in athymic nude mice
K Uetsuka; M Suzuki; C Kai; N Goto, In this experiment, we examined the protective effects of a novel quinone derivative, E3330, on MHV-2cc-induced chronic hepatitis in athymic nude mice for up to 3 weeks after virus infection. The daily dose of 25 mg/kg b.w. suppressed the viral replication in the liver and the progression of hepatic lesions. The expansion of small focal lesions at I week after viral inoculation (WAI) was suppressed at 2 WAI, and the lesions were still small at 3 WAI in E3330-administered group, whereas small focal lesions at I WAI were expanded at 2 WAI to fuse with each other at 3 WAI in the control group. E3330 therefore showed protective effects on MHV-2cc-induced chronic hepatitis in athymic nude mice, but further studies are needed to analyze the mechanism., JAPAN EXPER ANIMAL RES ASSN
EXPERIMENTAL ANIMALS, 1997年07月, ［査読有り］

Immunohistochemical study on galactosamine-induced subacute hepatitis in rats of JCL: Wistar-TGN (ARGHGEN) 1Nts strain (Mini rats)
K Uetsuka; M Suzuki; H Nakayama; K Doi, Immunohistochemical study was carried out on D-galactosamine hydrochloride (GalN)-induced subacute hepatitis in rats of JCL:Wistar-TGN (ARGHGEN) 1Nts strain (Mini rats), in which the expression of growth hormone gene is suppressed by the presence of an antisense transgene. Mini rats were given 1000 mg/kg of GalN once a week for 4 consecutive weeks and killed at 1, 2, 3 and 4 weeks after the first administration. At I week after the first administration, proliferation of small epithelial cells positive for both crfetoprotein and cytokeratin 7, i.e. so-called oval cells, was observed in the whole area of each hepatic lobule, and prominent deposition of fibronectin, laminin and type IV collagen was detected around these oval cells. Together with these extracellular matrix components, many activated Ito cells positive for both desmin and alpha-smooth muscle actin were obsereved With time, most of the oval cells formed duct-like structures and lost their positive stainability for alpha-fetoprotein, and many Ito cells became inactive. Deposition of fibronectin decreased rapidly from 2 weeks after the first administration. At 4 weeks after the first administration, deposition of laminin was detected only around the duct-like structures, where that of type IV collagen was also still prominent. These results suggest that a large population of oval cells differentiated into bile duct epithelial cells and that Ito cells and extracellular matrix components might play a role in this process., JAPAN EXPER ANIMAL RES ASSN
EXPERIMENTAL ANIMALS, 1997年07月, ［査読有り］

SCID-bg mice as xenograft recipients
S Shibata; T Asano; A Ogura; N Hashimoto; J Hayakawa; K Uetsuka; H Nakayama; K Doi, SCID-bg (scid/scid, beige/beige) is a strain of double-mutant mice with impaired lymphoid development and reduced natural killer (NK) cell activity. The present study was undertaken to evaluate the usefulness of SCID-bg mice as xenograft recipients. Fetal guineapig tissues (liver, thymus, spleen) were transplanted under the kidney capsule of the mice and their serum guineapig IgG levels were measured weekly thereafter. C.B.-17-scid and anti-asialo GM(1) antiserum-treated (NK-depleted) C.B.-17-scid (C.B.-17-scid-AGM(1)) mice that received the identical transplants were used as controls. Throughout the experimental period(1, 2, and 3 weeks after transplantation), the average serum guineapig IgG concentrations was highest in C.B.-17-scid-AGM(1) mice followed by SCID-bg mice and lowest in C.B.-17-scid mice without antiserum treatment, though we could not find any statistical significance among these groups. However, SCID-bg mice always showed the smallest within-group variance (individual difference) in the serum guineapig IgG concentrations (P<0.05, versus C.B.-17-scid-AGM(1) mice at 1,2, and 3 weeks and versus C.B.-17-scid mice at 2 weeks). The graft size was not significantly different among these three groups, but the spleen grafts in C.B.-17-scid mice contained fewer nucleate cells than the other two groups. These results indicate that the reduced NK cell activity by beige mutation is not crucial for the success of xenogenic transplantation, though SCID-bg mice may be useful as xenograft recipients with a consistent potential to retain the viability and function of engrafted tissues., ROYAL SOC MEDICINE PRESS LTD
LABORATORY ANIMALS, 1997年04月, ［査読有り］

Process of the development of T-2 toxin-induced apoptosis in the lymphoid organs of mice
J Shinozuka; GM Li; K Uetsuka; H Nakayama; K Doi, Female ICR:CD-1 mice orally treated with 10 mg/kg b.w. of T-2 toxin were killed at 1, 3, 6, 9, 12, 24 and 48 hr after treatment (HAT) and subjected to examination of the process of the development of T-2 toxin-induced apoptosis in the thymus and spleen. The early ultrastructural changes in lymphocytes characterized by shrinkage of the cell body and condensation of nuclear chromatin were detected at 3HAT in the thymus. The number of apoptotic lymphocytes observed by the in situ detection method for fragmented DNA increased drastically from 9 to 24 HAT in the thymus while it began to increase at 12 HAT in the spleen. The DNA ladder was first detected by agarose gel electrophoresis at 9 HAT and became clearer at 12 and 24 HAT in the thymus but was not clearly detected in the spleen throughout the observation period, Thus T-2 toxin-induced apoptosis developed earlier and was apparently severer in the thymus than in the spleen. Apoptois was first detected by electron microscopy, then by the in situ detection method for fragmented DNA, and finally by DNA agarose gel electrophoresis., JAPAN EXPER ANIMAL RES ASSN
EXPERIMENTAL ANIMALS, 1997年04月, ［査読有り］

T-2 toxin-induced apoptosis in lymphoid organs of mice.
Shinozuka J; Li GM; Kiatipattanasakul W; Uetsuka K; Nakayama H; Doi K
Experimental and Toxicologic Pathology., 1997年, ［査読有り］

T-2 toxin-induced apoptosis in intestinal crypt epithelial cells of mice
Guanmin Li; Junko Shinozuka; Koji Uetsuka; Hiroyuki Nakayama; Kunio Doi, The characteristics of T-2 toxin-induced cell damage in the intestinal crypt epithelia was investigated in mice. Following T-2 toxin-inoculation (0, 2.5, 5 and 10 mg/kg b.w.), dead cells showing pyknosis were sporadically observed in the crypt epithelia, and the nuclei of these cells were strongly stained by the modified TUNEL method which detects fragmented DNA in situ. Electron microscopically, the dead cells were characterized by shrinkage of the cell body and condensation of nuclear chromatin frequently along the nuclear membrane, and such nuclei were sometimes fragmented into small pieces. These morphological characteristics are well consistent with those of apoptosis. The mitotic index in the crypt epithelia drastically decreased at 6 hours after T-2 toxin-inoculation (6 HAI), but thereafter it recovered to almost the same value with that in control mice at 48 HAI. On the other hand, the apoptotic index in the crypt epithelia increased with the lapse of time. Clear mouse strain- and sex-differences were detected in the apoptotic index but not in the mitotic index. This is the first report that T-2 toxin caused apoptotic cell death in the intestinal crypt epithelial cells., Elsevier GmbH
Experimental and Toxicologic Pathology, 1997年, ［査読有り］

A case of canine nephroblastoma with unusual neuronal and mesenchymal components.　　　　　　　　　　　　　　　
Lu D-P; Taki R; Uetsuka K; Nakayama H; Doi K
Journal of Toxicologic Pathology., 1997年, ［査読有り］

The Role of Heparan Sulfate Proteoglycan, TGF-β1, and Kupffer Cells in Oval Cell Differentiation after Galactosamine (GaIN) Administration in Mini Rats
Koji Uetsuka; Michio Suzuki; Hiroyuki Nakayama; Kunio Doi, Six-week-old male Mini rats were intraperitoneally given 1,000 mg/kg of D-galactosamine hydrochloride (GaIN) once a week for 4 consecutive weeks and killed at 1, 2, 3, and 5 days (D) and 1, 2, 3, and 4 weeks (W) after the first GalN-administration, respectively. Proliferation of oval cells was observed at and after 3 D. Around them, prominent deposition of heparan sulfate proteoglycans was observed throughout the experimental period. Positive stainability for transforming growth facter (TGF)-β1 was detected in the cytoplasm of both bile duct epithelial cells in the Glisson's capsules and proliferating oval cells throughout the experimental period, and this may suggest an internalization of latent-type TGF-β1 in oval cells as well as bile duct epithelial cells. These results, in turn, indicate the differentiation of oval cells into bile duct epithelial cells. On the other hand, the number of Kupffer cells positive for ED2 peaked at 2D and thereafter decreased gradually, and no spacial relationship was detected between Kupffer cells and proliferating oval cells. © 1997, JAPANESE SOCIETY OF TOXICOLOGIC PATHOLOGY. All rights reserved.
Journal of Toxicologic Pathology, 1997年, ［査読有り］

Liver regeneration after partial hepatectomy in Mini rat.
Uetsuka K; Suzuki M; Ikeda A; Nakayama H; Doi K, Rats of Jcl : Wistar TGN (ARGHGEN) 1 Nts strain (Mini rats) are transgenic animals in which the expression of growth hormone (GH) gene is suppressed by the presence of an antisense transgene. In the present study, we examined hepatocyte regeneration after 2/3 partial hepatectomy (PH) in male Mini rats, using Wistar rats as controls. As a result, the restoration rate of liver weight was similar between the 2 strains, though bromodeoxyuridine (BrdU)- and proliferating cell nuclear antigen (PCNA)-labeling indices (LIs) of hepatocytes were generally higher in Wistar rats than in Mini rats at 1 day after PH. At 2 days, on the contrary to BrdU-LI, PCNA-LI was higher in Mini rats than in Wistar rats, and the difference in the total PCNA-LIs at 1 and 2 days between the 2 strains was not so large. This suggests that hepatocyte proliferation may start more slowly in Mini rats than in Wistar rats but the total proliferation rate during the regenerating process may not be significantly different between the 2 strains as a whole., JAPANESE SOCIETY OF TOXICOLOGIC PATHOLOGY
Journal of Toxicologic Pathology., 1997年, ［査読有り］

T-2 toxin-induced apoptosis in Peyer’s patches of mice.
Li GM; Shinozuka J; Uetsuka K; Nakayama H; Doi K, Lymphocytic apoptosis in Peyer's patches in the small intestine of mice orally inoculated with T-2 toxin (10 mg/ kg b.w.) were examined for up to 48 hours after inoculation (HAI). The number of apoptotic lymphocytes observed by in situ detection method of fragmented DNA increased significantly from 6HAI and reached a plateau at 24HAI. Ultrastructurally, lymphocytes characterized by shrinkage of the cell body and condensation of nuclear chromatin and/or margination of condensed chromatin along the nuclear membrane were observed from 6HAI. Some cell nuclei were fragmented into small pieces, and these small pieces were sometimes ingested by macrophages. © 1997, JAPANESE SOCIETY OF TOXICOLOGIC PATHOLOGY. All rights reserved.
Journal of Toxicologic Pathology., 1997年, ［査読有り］

Renal glomerular fibrosis in a cat
S Nakamura; S Shibata; K Shirota; K Abe; K Uetsuka; H Nakayama; N Goto; K Doi, Renal glomerular fibrosis was observed in a 1-year-old spayed female Japanese domestic cat that showed clinically advanced renal failure. In the glomeruli, increased homogeneous materials were stained strongly with aniline blue by Masson's trichrome and positive for anti-type III collagen antibody by immunohistochemical staining, causing mesangial sclerosis and capillary collapse. By electron microscopy, randomly arranged fibrils were observed in the expanded subendothelial and mesangial areas, and the fibrils showed periodicity characteristic of collagen fibers in longitudinal sections. These findings of glomerular lesions closely resemble those of human ''collagenofibrotic glomerulonephropathy,'' which has recently been described as a new type of glomerulonephropathy., AMER COLL VET PATHOLOGIST
VETERINARY PATHOLOGY, 1996年11月, ［査読有り］

Protective effect of recombinant interferon (IFN)-alpha/beta on MHV-2cc-induced chronic hepatitis in athymic nude mice
K Uetsuka; H Nakayama; N Goto, The protective effects of recombinant IFN-alpha/beta on MHV-2cc-induced chronic and persistent hepatitis in athymic nude mice were examined. The mice intraperitoneally (ip) inoculated with MHV-2cc at day 0 of experiment were divided into 4 groups. Three of them were administered io with recombinant IFN-alpha/beta at a daily dose of 1 x 10(3) IU from-1 (-1D-group), 0 (0D-group), and +1 day of experiment (+1D-group), respectively, for 3 consecutive weeks. The remaining one (control group) was given 0.1 ml/mouse of PBS from + 1 day of the experiment in the same way. Three mice in each group were killed at 1, 2 and 3 weeks post inoculation (WPI) with MHV, respectively. The liver virus titer in the control group increased gradually and maintained high levels throughout the experimental period. In the lFN-groups, particularly in the -1D- and 0D-groups, the virus titers were significantly lower than that in control group. Histopathologically, focal hepatic lesions were observed at 1 WPI and large irregular inflammatory lesions developed at 3WPI in the control group. Similar but somewhat less severe lesions were observed in the +1D-group. In the -1D- and 0D-groups, lesions were not observed at 1 WPI and only small organized lesions with mononuclear cell infiltration were seen at 3WPI. In conclusion, it was clarified in the present study that the progression of MHV-2cc-induced chronic hepatitis in athymic nude mice was effectively prevented by extrinsic IFN-alpha/beta when administered from -1 day and 0 day of the virus infection., JAPAN EXPER ANIMAL RES ASSN
EXPERIMENTAL ANIMALS, 1996年07月, ［査読有り］

Senile plaques in very aged cats
S Nakamura; H Nakayama; W Kiatipattanasakul; K Uetsuka; K Uchida; N Goto, Senile plaques were found in the cerebral cortices of three very aged cats (more than 18 years old). The plaques consisted of a coarse assembly of silver staining positive materials, and was morphologically different from the well-known classical, primitive, and diffuse plaques. Congophilic amyloid angiopathy was observed in a few cortical arterioles of the oldest cat (20 years old). The senile plaques and a few cortical blood vessels were immunopositive for amyloid beta-protein (A beta). A beta-positive materials were also sparsely distributed in the cortical neuropil but did not form senile plaques there. These findings should help to clarify the development of senile plaques and the early stage of A beta deposition., SPRINGER VERLAG
ACTA NEUROPATHOLOGICA, 1996年04月, ［査読有り］

Hepatitogenicity of three plaque purified variants of hepatotropic mouse hepatitis virus, MHV-2 in athymic nude mice
K Uetsuka; H Nakayama; N Goto, Hepatitogenicity of 3 plaque purified variants of hepatotropic mouse hepatitis virus, MHV-2 were examined in athymic BALB/c-nu/nu mice up to 9 weeks post infection (9WPI). All of the MHV-2S- and MHV-2M-infected mice died with severe acute hepatitis in 3WPI. On the other hand, MHI/-2L-infected mice did not die until 9WPI and showed signs of slow-developing chronic hepatitis with persistent infection under low serum virus neutralizing antibody titers. This suggests that MHV-2L-infected athymic nude mice may be useful as a new model of chronic Viral hepatitis., JAPAN EXPER ANIMAL RES ASSN
EXPERIMENTAL ANIMALS, 1996年04月, ［査読有り］

Ultrastructure of cardiac myocyte in the Asian elephant (Elephas maximus)
H Endo; TK Yamada; N Suzuki; G Suwa; K Uetsuka; O Hashimoto; M Kurohmaru; Y Hayashi, Cardiac myocytes of an Asian elephant (Elephas maximus) were observed by transmission electron microscopy. Typical ultrastructural features of cardiac myocytes are exhibited in the musculature of both the left and right atria, and left ventricle of the heart. Myofibrils, mitochondria, T-system and sarcoplasmic reticulum are well-developed within the cytoplasm. Many mitochondria are characteristically concentrated is some myocytes. Cardiac musculature is also distributed in the root of the caudal vena cava. Many atrial granules are detected not only in atrial myocytes, but also in the myocytes of the caudal vena cava. Atrial natriuretic polypeptide may be secreted from the caval venous wall in the elephant., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1995年12月, ［査読有り］

SENILE PLAQUES IN AN AGED 2-HUMPED (BACTRIAN) CAMEL (CAMELUS-BACTRIANUS)
S NAKAMURA; H NAKAYAMA; K UETSUKA; N SASAKI; K UCHIDA; N GOTO, Senile plaques with P-protein as a major constituent are a conspicuous feature in the brains of aged humans, monkeys, dogs, and bears. We found cerebral senile plaques of the diffuse and primitive type, but not the classical type, in an aged female camel of more than 20 years old. The senile plaques and a few cortical capillaries were immunoreactive with anti-beta-protein serum. Congophilic amyloid deposition was detected in a small number of the capillaries, but not in the senile plaques. We believe this to be the first detailed report of senile plaques in a herbivore, and these findings suggest the possibility of senile plaque formation in a wide variety of mammalian species., SPRINGER VERLAG
ACTA NEUROPATHOLOGICA, 1995年10月, ［査読有り］

Severe combined immunodeficiency (SCID) mouse hepatitis experimentally induced with low virulence mouse hepatitis virus
K. Uetsuka; H. Nakayama; N. Goto; K. Fujiwara, Springer New York LLC
Advances in Experimental Medicine and Biology, 1995年

COMPUTED-TOMOGRAPHY AND MAGNETIC-RESONANCE FINDINGS OF MENINGEAL SYNDROME IN A LEUKEMIC CAT
M MOROZUMI; N SASAKI; Y OYAMA; K UETSUKA; H NAKAYAMA; N GOTO, A 3-year-old Japanese domestic cat with a diagnosis of lymphocytic leukemia showed severe generalized seizures in the course of chemotherapy after leukemic condition was improved clinically. Computed tomography (CT) and magnetic resonance (MR) imaging of the brain were carried out. Both contrast procedures disclosed enhancements at the fair cerebri and the margin of cerebral cortex. Among these procedures contrast MR imaging demonstrated the lesion most clearly. Cytological examination of cerebrospinal fluid obtained by spinal puncture showed the infiltration of malignant cells and the diagnosis of meningeal syndrome associated with lymphocytic leukemia was defined. Intrathecal administration of cytosine arabinoside partially improved the neurologic dysfunction. Autopsy and histopathological examination confirmed the infiltration of leukemic cells in the areas of meningeal lesion demonstrated with contrast CT and MR imaging. Thus these imaging techniques, especially contrast MR imaging, are useful tools for rapid and precise diagnosis of meningeal syndrome., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1993年12月, ［査読有り］

SYSTEMIC CEROID-LIPOFUSCINOSIS IN A JAPANESE DOMESTIC CAT
H NAKAYAMA; K UCHIDA; T SHOUDA; K UETSUKA; N SASAKI; N GOTO, An 11-month-old castrated male Japanese domestic cat was euthanized because of neurological symptoms such as shivering and difficulty of walking. Histopathological examination showed glial proliferation and marked deposition of ceroid-lipofuscin in the neuronal and glial cells of the brain. Ceroid-lipofuscin was deposited also in reticuloendothelial cells of the liver, spleen and some lymph nodes., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1993年10月, ［査読有り］

GLOMERULOPATHY WITH IGA DEPOSITION IN THE DOG
Y MIYAUCHI; H NAKAYAMA; K UCHIDA; K UETSUKA; A HASEGAWA; N GOTO, In 100 dogs autopsied, glomerular IgA deposition was examined by the immunofluorescence technique and the histopathological features of glomeruli with IgA deposition were examined by light and electron microscopy. The incidence of the IgA deposition was age-related but there were no sex and breed predisposition. Deposition of IgA was observed mainly in mesangial areas in approximately a half (47%) of dogs examined. IgG, IgM and C3 often co-deposited. Histopathology of the glomeruli with IgA deposition indicated increase of mesangial cells, crescent formation, hemispherical deposits in paramesangial areas and glomerular sclerosis. Ultrastructurally electron dense substances positive for IgA deposited in mesangial and paramesangial areas. The examination to know the relation between the severity of IgA deposition and the number of mesangial cells or percent of the cells to total glomerular cells indicated that mesangial cells increased at the early stage of the disease and subsequently epithelial and endothelial cells proliferated as the increasing amount of IgA. Dogs suffering from enteritis or liver diseases showed high incidence of glomerular IgA deposition., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1992年10月, ［査読有り］

3 CASES OF FELINE SCLEROSING LYMPHOCYTIC CHOLANGITIS
H NAKAYAMA; K UCHIDA; SK LEE; K UETSUKA; A HASEGAWA; N GOTO, JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1992年08月, ［査読有り］

IMMUNOHISTOCHEMICAL STUDIES ON CANINE CEREBRAL AMYLOID ANGIOPATHY AND SENILE PLAQUES
K UCHIDA; Y TANI; K UETSUKA; H NAKAYAMA; N GOTO, Amyloid protein was isolated from the cerebral meninges of 4 aged dogs with cerebral amyloid angiopathy. By immunoblot analysis, antiserum against synthetic oligo-peptide consisting of 1-28 amino acid of amyloid beta protein recognized prominent wide band ranging from 14 to 18 kilodalton (kd). When amyloid samples were solubilized by formic acid, the antiserum recognized lower molecular weight band ranging from 3 to 4 kd. Immunohistochemical studies on cerebral amyloid angiopathy and senile plaques were performed in 17 aged dogs. Anti-amyloid beta protein serum labeled amyloid deposits in cerebral vessel walls and senile plaques. Compact deposits of beta protein were detected in primitive or classical plaques. After using formic acid pretreatment, diffuse deposits of beta protein in the neuropil representing diffuse plaques were detectable. Classical and primitive plaques reacted with antiserum against glial fibrillary acidic protein, while not with antisera against alpha 1-antichymotrypsin, IgG and IgM. Amyloid deposits in the intestines of aged dogs examined, did not react with anti-amyloid beta protein serum., JAPAN SOC VET SCI
JOURNAL OF VETERINARY MEDICAL SCIENCE, 1992年08月, ［査読有り］

Mechanisms of mycotoxin-induced dermal toxicity and tumorigenesis through oxidative stress-related pathways
Kunio Doi; Koji Uetsuka
Journal of Toxicologic Pathology, 2014年

Mechanisms of Mycotoxin-Induced Neurotoxicity through Oxidative Stress-Associated Pathways
Kunio Doi; Koji Uetsuka
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2011年08月, ［査読有り］

動物病理学総論（第４版）
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文永堂出版, 2023年03月
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動物病理学各論（第３版）
日本獣医病理学専門家協会; 相原, 直之
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動物の衛生 第２版　　　　　　　　　　　　　　　
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動物病理カラーアトラス 第２版　　　　　　　　　　　　　　　
日本獣医病理学専門家協会 編, 共著
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動物病理学総論（第３版）　　　　　　　　　　　　　　　
共著
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新獣医学辞典　　　　　　　　　　　　　　　
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動物病理カラーアトラス　　　　　　　　　　　　　　　
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ストランディングフィールドガイド 海の哺乳類　　　　　　　　　　　　　　　
共訳
海游舎, 1996年08月10日
9784905930747

飼育下にある哺乳類、鳥類、爬虫類から分離したEnterococcus faecalis株の系統解析　　　　　　　　　　　　　　　
坂田 友希; 成廣 隆; 上塚 浩司
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佐藤 洸士郎、里見 峻、成廣 隆、上塚 浩司
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里見 峻、佐藤 洸士郎、成廣 隆、上塚 浩司
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日本の野鳥由来腸球菌 Enterococcus faecalis の抗菌薬耐性調査　　　　　　　　　　　　　　　
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日本環境感染学会 第38回大会（パシフィコ横浜）, 2023年07月22日
20230720, 20230722

茨城大学阿見キャンパスにおけるヒヨドリ（Hypsipetes amaurotis）の腸内細菌叢の16S rRNA遺伝子アンプリコンシーケンス解析　　　　　　　　　　　　　　　
君和田 勁、成廣 隆、島 綾香、會津 光博、杉浦 奈都子、上塚 浩司
2021年 日本バイオインフォマティクス学会年会,第10回 生命医薬情報学連合大会, 2021年09月28日, 浜田道昭（早稲田大学理工学術院、産業技術総合研究所、日本医科大学）
20210927, 20210929

腸内細菌叢は宿主動物の健康状態で変化するか？ ー日立市かみね動物園での検証ー　　　　　　　　　　　　　　　
坂場 円香、成廣 隆、飯田 伸弥、生江 信孝、上塚 浩司
2021年 日本バイオインフォマティクス学会年会,第10回 生命医薬情報学連合大会, 2021年09月28日, 浜田道昭（早稲田大学理工学術院、産業技術総合研究所、日本医科大学）
20210927, 20210929

Comparative microbial community analysis of the intestinal flora in wild birds captured at Fukushima-gata lake, Niigata, Japan.　　　　　　　　　　　　　　　
Uetsuka K; Shimizu T; Shima A; Ishihara G; Morimoto G; Deguchi T; Narihiro T
日本バイオインフォマティクス学会 2019年年会,第8回 生命医薬情報学連合大会, 2019年09月10日, 日本バイオインフォマティクス学会、日本オミックス学会

食性の異なる各種動物からのウェルシュ菌の分離調査　　　　　　　　　　　　　　　
清水かな、上塚浩司
日本家畜衛生学会 第90回大会, 2019年07月20日, 日本家畜衛生学会

腸内のウエルシュ菌量の死後変化についての検討モデルの開発（２）　　　　　　　　　　　　　　　
石塚由佳、上塚浩司
日本家畜衛生学会 第89回大会, 2018年12月14日, 日本家畜衛生学会

茨城大学阿見キャンパス内で捕獲した野生ネズミでのウエルシュ菌（Clostridium perfingens）保有調査
〇関 光一朗、成廣 隆、上塚浩司
日本微生物生態学会 第32回大会, 2018年08月12日, 日本微生物生態学会

野生カラスの腸内容物からのClostridium perfringensの分離および分離菌株の遺伝学的解析　　　　　　　　　　　　　　　
鈴木菜月、成廣 隆、〇上塚浩司
日本家畜衛生学会 第88回大会, 2018年07月07日, 日本家畜衛生学会

競争用馬の感染性とみられる皮膚炎からの細菌・真菌の分離同定と薬剤感受性調査　　　　　　　　　　　　　　　
○牛屋 重人、上塚 浩司
日本家畜衛生学会 第87回大会, 2017年12月15日, 日本家畜衛生学会

カラス、ムクドリ、スズメが腸内に保有するウエルシュ菌の調査　　　　　　　　　　　　　　　
鈴木菜月、永井あゆみ、仲本あゆ美、〇上塚浩司
第４回日本獣医病理学専門家協会学術集会, 2017年03月30日, 日本獣医病理学専門家協会

スズメの腸内容物からのCW卵黄寒天培地を用いたウエルシュ菌分離　　　　　　　　　　　　　　　
山本有花、上塚浩司
日本家畜衛生学会第85回大会, 2016年12月02日

野生ネズミの腸内容物と糞でのウエルシュ菌の菌量の対応　　　　　　　　　　　　　　　
関 光一朗、上塚 浩司
日本家畜衛生学会 第85回大会, 2016年12月02日

腸内のウエルシュ菌量の死後変化についての検討モデルの開発－Sprague Dawleyラットを用いた検討　　　　　　　　　　　　　　　
石塚由佳、上塚浩司
日本家畜衛生学会 第85回大会, 2016年12月02日

茨城県内の留鳥の腸内容物からのウエルシュ菌（Clostridium perfringens）の分離頻度、分離菌量、毒素型の調査　　　　　　　　　　　　　　　
鈴木菜月、仲本あゆ美、永井あゆみ、上塚浩司
第159回日本獣医学会学術集会, 2016年09月07日

高齢の台湾小耳種豚でみられた副腎髄質の増殖性病変の病理学的解析　　　　　　　　　　　　　　　
上塚浩司、鈴木敬之、富岡ひとみ、チェンバーズ ジェームズ、内田和幸、中山裕之、土井邦雄、布谷鉄夫
第154回日本獣医学会学術集会, 2012年09月

ガチョウの腹腔内腫瘤（診断：ガチョウの悪性セルトリ細胞腫）　　　　　　　　　　　　　　　
上塚浩司
第51回獣医病理学研修会, 2011年09月

新生豚にみられた先天性過骨症の一例　　　　　　　　　　　　　　　
第148回日本獣医学会学術集会, 2009年09月

豚の回腸（診断：肉芽腫性回腸炎および壊死性出血性結腸炎）　　　　　　　　　　　　　　　
上塚浩司
第49回獣医病理学研修会, 2009年04月

ネコの脂肪肝における伊東細胞の形態　　　　　　　　　　　　　　　
上塚浩司、坂井学、磯部香里、中山裕之
第144回日本獣医学会学術集会, 2007年09月

Histopathological characteristics of hepatic lipogranulomatous lesions in dogs with portosystemic shunt　　　　　　　　　　　　　　　
Uetsuka K; Isobe K; Doi K; Nakayama H
The American College of Veterinary Pathologists (ACVP): The 57th Annual Meeting, 2006年12月

ブタとウシの肝の伊東細胞とクッパー細胞に関する免疫組織化学的性状の解析　　　　　　　　　　　　　　　
上塚浩司、西川智美、中山裕之
第142回日本獣医学会学術集会, 2006年09月

伴侶動物の肝臓脂肪代謝病の病理　　　　　　　　　　　　　　　
上塚浩司
第141回日本獣医学会学術集会,日本獣医病理学会スライドセミナー, 2006年03月

肝再生の可能性：実験病理学的見地から　　　　　　　　　　　　　　　
上塚浩司
第141回日本獣医学会学術集会,臨床分科会シンポジウム, 2006年03月

ネコの肝の伊東細胞とクッパー細胞は特有の特徴を有する　　　　　　　　　　　　　　　
上塚浩司、西川智美、八十島昭、中山裕之、土井邦雄
第140回日本獣医学会学術集会, 2005年09月, 日本獣医学会

db/dbマウスの肝再生障害における非実質細胞の関与についての検討　　　　　　　　　　　　　　　
上塚浩司、山内啓史、白井真人、中山裕之、土井邦雄
第52回日本実験動物学会学術集会, 2005年06月, 日本実験動物学会

マウス3分の2肝部分切除後の肝再生に対する血管新生阻害薬FR118487投与の影響　　　　　　　　　　　　　　　
上塚浩司、中山裕之、土井邦雄
第138回日本獣医学会学術集会, 2004年09月, 日本獣医学会

肝再生における類洞内皮細胞増殖の意義 －脂肪肝モデルと血管新生阻害薬モデル－　　　　　　　　　　　　　　　
上塚浩司
第11回肝細胞研究会, 2004年07月, 肝細胞研究会

Angiopoietin発現アデノウイルスベクターの開発　　　　　　　　　　　　　　　
上塚浩司、中山裕之、土井邦雄
第137回日本獣医学会学術集会, 2004年04月, 日本獣医学会

db/dbマウスにおける肝再生障害メカニズムの探索 (2) －類洞血管からのアプローチ－　　　　　　　　　　　　　　　
上塚浩司、山内啓史、中山裕之、土井邦雄
第136回日本獣医学会学術集会, 2003年10月, 日本獣医学会

3分の2部分切除後のマウス肝再生における類洞内皮細胞についての基礎的解析　　　　　　　　　　　　　　　
上塚浩司
第10回肝細胞研究会, 2003年07月, 肝細胞研究会

肝臓stem cellのin vitroモデルであるRLE細胞を用いたTet-Off Systemでのc-kit/SCFシグナル系の役割の解析　　　　　　　　　　　　　　　
上塚浩司、中山裕之、土井邦雄、Snorri.S.Thorgeirsson
第48回日本実験動物学会学術集会, 2001年05月, 日本実験動物学会

2023年08月, 日本微生物資源学会

2023年04月, 日本乳酸菌学会

2019年04月, 日本バイオインフォマティクス学会

2018年04月, 日本微生物生態学会

2018年04月, 腸内細菌学会

2016年08月, 日本家畜衛生学会

2015年04月, 日本細菌学会

2014年04月, ヒトと動物の関係学会

2008年11月, 人と動物の共通感染症研究会

2008年10月, 鶏病研究会

2008年06月, 日本豚病研究会

2001年10月, 日本獣医病理学専門家協会

1992年03月, 日本獣医学会

農場で発生する腸炎にカラス由来のウエルシュ菌はどのように関与しているか？ －MLST法による遺伝学的解析での検討－　　　　　　　　　　　　　　　
2017年04月 - 2018年03月

βアミロイド沈着と脳老化の比較生物学的研究
基盤研究(B)
東京大学
2005年 - 2007年

母体-胎盤-胎児軸における胎児毒性の発現機構に関する多面的解析
基盤研究(B)
東京大学
2004年 - 2006年

タイ国における飼料中のかび毒自然汚染の家畜家禽疾病発生に及ぼす影響に関する研究
基盤研究(B)
東京大学
2003年 - 2005年

病態のDNAメチル化解析
基盤研究(A)
東京大学
2003年 - 2005年

老齢動物における痴呆関連遺伝子発現についての比較進化医学的研究
基盤研究(B)
東京大学
2002年 - 2004年

環境化学物質と紫外線による皮膚障害の性状と発現機序に関する多面的解析
基盤研究(B)
東京大学
1998年 - 2000年

oval cellの増殖・分化と細胞外基質に関する基礎的研究
奨励研究(A)
東京大学
1998年 - 1999年

神経細胞の発生および老化におけるアポトーシスとDNAメチル化の機序
基盤研究(C)
東京大学
1997年 - 1998年

環境化学物質の皮膚刺激性ならびに紫外線との複合作用に関する基礎的研究
萌芽的研究
東京大学
1997年 - 1997年

病態動物における薬物の毒性発現様式の修飾機構に関する基礎的研究
萌芽的研究
東京大学
1996年 - 1996年

癌原遺伝子C-maf産物タンパク質のニワトリ網膜における発現とその機能
一般研究(C)
東京大学
1995年 - 1995年

人の各型慢性肝炎に対応する動物ウイルス肝炎モデルの開発と応用
一般研究(C)
東京大学
1994年 - 1994年