2022/11/22 更新

写真a

タカハシ マサシ
高橋 雅
TAKAHASHI MASASHI
所属
農水産獣医学域獣医学系 共同獣医学部 附属動物病院 准教授
職名
准教授

学位

  • 博士(獣医学) ( 2014年9月   山口大学 )

研究キーワード

  • 血液疾患

  • 腫瘍

  • 血栓

経歴

  • 鹿児島大学   農水産獣医学域獣医学系 共同獣医学部 附属動物病院   准教授

    2018年10月 - 現在

  • 鹿児島大学   農水産獣医学域獣医学系 共同獣医学部 獣医学科   助教

    2015年5月 - 2018年9月

所属学協会

  • 日本獣医がん学会

    2015年10月 - 現在

 

論文

  • FURUSAWA Yu, TAKAHASHI Masashi, SHIMA-SAWA Mariko, HATAI Hitoshi, MIYOSHI Noriaki, YAMATO Osamu, YABUKI Akira .  Immunocytochemical evaluation of epithelial–mesenchymal transition in epithelial tumors of dogs and cats .  The Journal of Veterinary Medical Science83 ( 9 ) 1363 - 1368   2021年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本獣医学会  

    <p>Epithelial–mesenchymal transition (EMT) plays a crucial role in metastasis of epithelial tumors; however, it is challenging to detect EMT by cytology. In the present study, EMT was visualized by fluorescence-immunocytochemistry (FICC). Air-dried smears from epithelial tumors of dogs (n=22) and cats (n=9) were stained using mouse monoclonal anti-E-cadherin and rabbit monoclonal anti‐vimentin antibodies. Enzymatic immunohistochemistry (IHC) revealed that 51.6% (8/22 in dogs, 8/9 in cats) of the cases showed EMT. In dogs, FICC could detect EMT in 62.5% (5/8) of those cases. In cats, FICC could detect EMT in 100% (8/8) of the cases. In conclusion, the present FICC method could successfully detect EMT using conventional air-dried cytology smear slides.</p>

    DOI: 10.1292/jvms.21-0021

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  • Furusawa Y, Takahashi M, Shima-Sawa M, Yamato O, Yabuki A. .  Argyrophilic nucleolar organizer regions staining for cytology smears in dogs and cats. .  J Vet Med Sci.24 ( 82 ) 1267 - 1270   2020年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本獣医学会  

    <p>The argyrophilic nucleolar organizer regions (AgNORs) are cellular proliferation markers, crucial for predicting the clinical course and aggressiveness of tumors. The purpose of this study was to establish an easy and practical AgNOR staining method in the cytology of dogs and cats. Air-dried cytological slides were prepared from dogs (n=14) and cats (n=12). Acetone, formalin, ethanol and methanol were tested as fixatives for AgNOR staining. Subsequently, various methods of Romanowsky-based counterstains were tested before and after AgNOR staining. Clear and strong AgNOR spots were observed with all fixatives, and post-May–Grünwald staining was the best counterstaining method. The established method showed clear AgNOR spots even in the long-term storage samples and Romanowsky-stained ones.</p>

    DOI: 10.1292/jvms.20-0271.

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  • Furusawa Y, Takahashi M, Iwanaga T, Yabuki A, Kano R, Hatai H, Sueno W, Endo Y, Momoi Y. .  Systemic Inonotus sp. Infection in a dog. .  J Vet Med Sci.24 ( 82 ) 779 - 783   2020年6月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本獣医学会  

    <p>A 3-years-old male golden retriever was presented for decreased activity (lethargy), anorexia, and titubation. Superficial lymph nodes were enlarged, and arrhythmia and tachycardia were auscultated. Fungal hyphae-like structures were detected in the biopsy samples from an enlarged lymph node and spleen. Nucleotide sequence of the internal transcribed spacer region of the fungi amplified by PCR was highly homologous to that of <i>Inonotus pachyphloeus</i>. The dog was treated with antifungal agents such as itraconazole, fluconazole, and voriconazole. Clinical signs resolved for 325 days but the dog died suddenly, possibly because of arrhythmia. Postmortem examination revealed the presence of a disseminated fungal infection. This report describes the case of canine systemic <i>Inonotus</i> sp. infection treated by an antifungal agent.</p>

    DOI: 10.1292/jvms.19-0455.

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  • Kato D, Takahashi M, Yonezawa T, Ohmi A, Takeda S, Nakagawa T, Hosoda S, Kanemoto H, Fukushima K, Ohno K, Matsuki N, Tsujimoto H .  Evaluation of an automated point-of-care test system for measuring thrombin-antithrombin complex in dogs. .  Journal of veterinary emergency and critical care (San Antonio, Tex. : 2001)30 ( 1 ) 102 - 106   2020年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1111/vec.12913

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  • Tomita A. .  Development of canine X-chromosome inactivation pattern analysis for the detection of cell clonality by incorporating the examination of the SLIT and NTRK-like family member 4 (SLITRK4) gene .  Research in Veterinary Science125   170 - 175   2019年8月

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    記述言語:日本語   出版者・発行元:Research in Veterinary Science  

    DOI: 10.1016/j.rvsc.2019.06.004

    Scopus

  • Motegi T, Tomiyasu H, Goto-Koshino Y, Takahashi M, Hiyoshi-Kanemoto S, Fujino Y, Ohno K, Tsuimoto H. .  Prognostic value of CD44 variant isoform expression in dogs with multicentric high-grade B-cell lymphoma. .  Am J Vet Res   2018年9月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

  • Sato M, Mochizuki H, Goto-Koshino Y, Fujiwara-Igarashi A, Takahashi M, Ohno K, Tsujimoto H. .  Prognostic significance of hypermethylation of death-associated protein kinase (DAPK) gene CpG island in dogs with high-grade B-cell lymphoma. .  Vet Comp Oncol.16 ( 3 ) 409 - 415   2018年3月査読

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1111/vco.12395

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  • Yamazaki Hiroki, Miura Naoki, Lai Yu-Chang, Takahashi Masashi, Goto-Koshino Yuko, Yasuyuki Momoi, Nakaichi Munekazu, Tsujimoto Hajime, Setoguchi Asuka, Endo Yasuyuki .  Effects of toceranib phosphate (Palladia) monotherapy on multidrug resistant lymphoma in dogs. .  J Vet Med Sci79 ( 7 ) 1225 - 1229   2017年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本獣医学会  

    We examined whether multidrug resistant (MDR) canine lymphoma increases gene expression for platelet-derived growth factor receptor alpha (PDGFRalpha), vascular endothelial growth factor receptor 2 (VEGFR2), and c-KIT, and whether toceranib phosphate (TOC) has potential as a treatment for MDR canine lymphoma. The clinical data showed that PDGFRalpha expression was higher in canine subjects with MDR T-cell lymphoma than in those with untreated T-cell lymphoma, and that c-KIT expression was greater in subjects with T-cell lymphoma than in those with B-cell lymphoma. TOC monotherapy was well tolerated without serious adverse effects, and two of the five subjects that received TOC exhibited partial responses. The data presented here could contribute to the assessment of TOC-based therapy for dogs with MDR or T-cell lymphoma.

    DOI: 10.1292/jvms.16-0457

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  • Jikuya Mao, Tateno Morihiro, Takahashi Masashi, Endo Yasuyuki .  The presence of tick-borne diseases in domestic dogs and cats living on Iriomote-jima and Tsushima islands. .  J Vet Med Sci79 ( 6 ) 1086 - 1090   2017年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:公益社団法人 日本獣医学会  

    The Iriomote cat and Tsushima leopard cat are endangered wildcats in Japan and inhabit only Iriomote-jima and Tsushima islands, respectively. Domestic dogs and cats living on Iriomote-jima and Tsushima islands were surveyed to clarify the interrelationship between wildcats and domestic animals regarding tick-borne disease transmission. Pathogen-derived DNA in blood samples was detected by polymerase chain reaction. Babesia gibsoni was detected in dogs of Iriomote-jima, and Hepatozoon felis and hemoplasmas were detected in domestic cats of Tsushima. Because the H. felis detected in this study was closely related to that isolated from wildcats, we suspect that common H. felis is harbored and transmitted among wildcats and domestic cats via ticks in Tsushima.

    DOI: 10.1292/jvms.16-0546

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  • Tamamoto T, Ohno K, Takahashi M, Fukushima K, Kanemoto H, Fujino Y, Tsujimoto H .  Plasma matrix metalloproteinase-9 activity in cats with lymphoma. .  Vet Comp Oncol15 ( 1 ) 1 - 5   2017年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In this study, plasma MMP-9 activity was evaluated in cats with lymphoma. Plasma samples were obtained from 26 cats with lymphoma before treatment. From 13 of the included 26 cats, plasma samples were obtained 4 weeks after the initiation of treatment. Plasma samples were also obtained from 10 healthy cats as a control. Plasma MMP-9 activity was examined by gelatin zymography and semi-quantitative value (arbitrary unit; a.u.) for each sample was calculated. Relatively high levels of MMP-9 were observed in cats with lymphoma compared with those in healthy control cats. MMP-9 quantification through zymography showed significantly higher activity in cats with lymphoma (median, 0.63 a.u.; range, 0.23-3.24 a.u.) than in healthy controls (0.22 a.u.; 0.12-0.46 a.u.; P < 0.01). MMP-9 activities were significantly different before (0.73 a.u.; 0.30-3.24 a.u.) and after treatment (0.50 a.u.; 0.14-1.32 a.u.; P = 0.017). Measuring plasma MMP-9 activity in cats with lymphoma may become an appropriate monitoring tool for feline lymphoma.

    DOI: 10.1111/vco.12097

    PubMed

  • Sato Toru, Ohno Koichi, Tamamoto Takashi, Oishi Mariko, Kanemoto Hideyuki, Fukushima Kenjiro, Goto-Koshino Yuko, Takahashi Masashi, Tsujimoto Hajime .  Assesment of severity and changes in C-reactive protein concentration and various biomarkers in dogs with pancreatitis. .  J Vet Med Sci79 ( 1 ) 35 - 40   2017年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Canine pancreatitis is a relatively common disorder, and its mortality rate remains high. However, prognostic factors for pancreatitis based on evidence are limited. Moreover, the relationship between changes in C-reactive protein (CRP) concentration-an important prognostic factor for human patients with acute pancreatitis-and the prognosis of dogs with pancreatitis has not been widely studied. Therefore, we examined prognostic factors for canine pancreatitis during the first medical examination and evaluated the usefulness of serial CRP measurements during hospitalization. Sixty-five dogs met the inclusion criteria, including 22 that were hospitalized and treated. In Study 1, a multivariate analysis revealed that three factors- decreased platelet count and a marked (greater than 1,000 microg/l) elevation of specific canine pancreatic lipase (Spec cPL) concentration at the first medical examination, as well as elevated blood urea nitrogen (BUN) and/or creatinine (CRE) level-were significantly different between the survivors and nonsurvivors. Moreover, CRP concentrations on the third and fourth days were significantly different between the two groups in Study 2. An evaluation of the decreased platelet count, remarkable elevation of Spec cPL concentration at the first medical examination, elevation of BUN and/or CRE as well as serial CRP concentration measurements may be useful for predicting the prognosis of canine pancreatitis.

    DOI: 10.1292/jvms.16-0009

    PubMed

  • Kato Daiki, Takahashi Masashi, Harada Tomoyuki, Tsuboi Masaya, Uchida Kazuyuki, Fukushima Kenjiro, Kanemoto Hideyuki, Fujino Yasuhito, Ohno Koichi, Nakayama Hiroyuki, Tsujimoto Hajime .  Intranuclear inclusions in a dog with B-cell leukemia. .  Vet Clin Pathol45 ( 4 ) 579 - 583   2016年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A 7-year-old Shetland Sheepdog was presented with anorexia. A CBC indicated thrombocytopenia and neutropenia. Bone marrow cytology revealed that 67.7% of all nucleated cells (ANC) were anaplastic large mononuclear cells. These cells were confirmed to be of B-cell origin based on IgH rearrangement, immunohistochemical, and flow cytometric analysis. Microscopic examination revealed that the neoplastic cells had intranuclear inclusions resembling Dutcher bodies. Immunohistochemistry confirmed that the intranuclear inclusions were immunopositive for IgG antibodies. The periodic acid-Schiff reaction was negative for the presence of polysaccharides and related substances. Although the dog achieved complete remission with a multi-drug chemotherapy protocol, it ultimately died because of tumor progression and acute renal insufficiency on day 201. This is the first known case of canine acute B-cell leukemia with intranuclear inclusions resembling Dutcher bodies.

    DOI: 10.1111/vcp.12420

    PubMed

  • Mizutani Noriyuki, Goto-Koshino Yuko, Takahashi Masashi, Uchida Kazuyuki, Tsujimoto Hajime .  Clinical and histopathological evaluation of 16 dogs with T-zone lymphoma. .  J Vet Med Sci78 ( 8 ) 1237 - 1244   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Clinical and histopathological characteristics of 16 dogs with nodal paracortical (T-zone) lymphoma (TZL) were evaluated. At initial examination, generalized lymphadenopathy was found in all dogs, and peripheral lymphocytosis was found in 10 of the 16 dogs. At initial diagnosis or during the disease course, 8 dogs (50%) were affected with demodicosis. Immunohistochemical analysis for CD3, CD20 and CD25 was performed for 6 dogs with TZL; the tumor cells were positive for CD3 and CD25 and negative for CD20. Median overall survival time was 938 days. A watchful waiting approach was adopted for 6 cases (38%), and 5 of the 6 dogs were still alive at the end of follow-up. The clinical course of TZL in dogs is generally indolent; however, many cases develop a variety of infectious and other neoplastic diseases after the diagnosis of TZL.

    DOI: 10.1292/jvms.15-0688

    PubMed

  • Sato Masahiko, Yamazaki Jumpei, Goto-Koshino Yuko, Setoguchi Asuka, Takahashi Masashi, Baba Kenji, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Minimal residual disease in canine lymphoma: An objective marker to assess tumour cell burden in remission. .  Vet J215   38 - 42   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Lymphoma is the most common haematopoietic malignancy in dogs. Since a high proportion of dogs with lymphoma achieve remission soon after initiation of chemotherapy, an objective marker assessing treatment efficacy is required. Following clinical remission, the residual population of tumour cells can be referred to as the minimal residual disease (MRD). MRD traditionally has been detected by cytology and flow cytometry; however, if the burden of malignant cells is low, these methods might not be sufficiently sensitive to detect MRD. As an extension of the development of PCR for antigen receptor gene rearrangements (PARR) in dogs, there has been recent progress in the application of real-time quantitative PCR (RT-qPCR) to canine lymphoma. With the RT-qPCR system, a very high sensitivity (1 cell per 10,000 cells) has been achieved by preparing allele-specific oligonucleotide primers and probes designed from neoplastic clones of each dog. A series of MRD diagnostics studies employing the RT-qPCR system has revealed its usefulness as a prognostic indicator, an objective marker of treatment efficacy and a predictor of relapse for dogs with lymphoma receiving chemotherapy. Introduction of the MRD monitoring system will provide an innovative scientific tool in the development of superior treatments and monitoring strategies for canine lymphoma.

    DOI: 10.1016/j.tvjl.2016.05.012

    PubMed

  • Hiyoshi-Kanemoto Saaya, Goto-Koshino Yuko, Fukushima Kenjiro, Takahashi Masashi, Kanemoto Hideyuki, Uchida Kazuyuki, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Detection of circulating tumor cells using GeneScan analysis for antigen receptor gene rearrangements in canine lymphoma patients. .  J Vet Med Sci78 ( 5 ) 877 - 881   2016年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The presence of circulating tumor cells (CTCs) serves as a prognostic marker and indicator of disease relapse, as well as a means of evaluating treatment efficacy in human and canine lymphoma patients. As an extension of our previous study for the construction of clinically useful GeneScan system, we utilized the GeneScan system for detecting CTCs in canine lymphoma patients. Samples from the primary lesion and peripheral blood mononuclear cells (PBMCs) were obtained from 32 dogs with lymphoma at initial diagnosis. All samples were subjected to polymerase chain reaction (PCR) for antigen receptor gene rearrangements (PARR) followed by GeneScan analysis. Common clonal rearrangements with identical amplified fragments were detected in both the primary lesion and PBMCs in 19 of the 32 dogs (59.4%). However, the detection rate of CTCs varied among the anatomical classification of lymphoma studied. GeneScan analysis following PARR would facilitate studies on determining the clinical significance of CTCs in canine lymphoma patients.

    DOI: 10.1292/jvms.15-0433

    PubMed

  • Fukushima Kenjiro, Eguchi Nozomi, Ohno Koichi, Kanemoto Hideyuki, Takahashi Masashi, Igarashi Hirotaka, Ohmi Aki, Nakashima Ko, Tsujimoto Hajime .  Efficacy of leflunomide for treatment of refractory inflammatory colorectal polyps in 15 Miniature Dachshunds. .  J Vet Med Sci78 ( 2 ) 265 - 269   2016年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Inflammatory colorectal polyp (ICRP), common in miniature dachshunds, presents with hematochezia, tenesmus and mucoid feces. Although an 80% response rate has been reported when treated with prednisolone and cyclosporine, effective treatment is needed for the remaining 20% of ICRP dogs. Leflunomide is an immunosuppressive drug reported as effective in several immune-mediated diseases. In the present study, we retrospectively evaluated the efficacy and adverse effects of leflunomide in 15 ICRP dogs that were refractory to treatment with prednisolone and cyclosporine. Treatment efficacy was assessed by endoscopy, clinical symptoms and rectal palpation. Adverse effects were determined by clinical symptoms and blood testing during follow-up. The leflunomide treatment response rate was 93.3%. The median dosage of leflunomide and the median response time were 3 mg/kg (range: 1.7-4.0 mg/kg) and 35 days (range: 20-119 days), respectively. Adverse effects observed included lethargy (3 dogs), anorexia (1 dog), respiratory symptoms (1 dog), leukocytopenia (2 dogs), thrombocytopenia (1 dog), anemia (1 dog) and liver enzyme elevation (8 dogs). Most of the adverse effects improved with symptomatic treatment and leflunomide discontinuation or dosage reduction. In conclusion, leflunomide treatment is effective in ICRP dogs refractory to treatment with prednisolone and cyclosporine. Because several adverse effects were observed, close monitoring is needed during leflunomide treatment follow-up.

    DOI: 10.1292/jvms.15-0129

    PubMed

  • Fujiwara-Igarashi Aki, Igarashi Hirotaka, Mizutani Noriyuki, Goto-Koshino Yuko, Takahashi Masashi, Ohno Koichi, Tsujimoto Hajime .  Expression profile of circulating serum microRNAs in dogs with lymphoma. .  Vet J205 ( 2 ) 317 - 321   2015年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Serum microRNAs (miRNAs) are mediators of cell-to-cell communication and alter the cellular microenvironment; they are stable for hours under certain conditions in body fluids despite the presence of RNases. Certain miRNAs have been found to be altered in the serum or plasma of humans with various cancers and may represent promising, non-invasive biomarkers for various diseases in humans and animals. The objective of this study was to determine the expression profile of circulating miRNAs in the serum of dogs with lymphoma. Serum samples were obtained from 61 dogs with lymphoma and 40 control dogs, and real-time reverse transcription-polymerase chain reaction was used for miRNA measurement. In order to select candidate genes, a comprehensive expression analysis was undertaken prior to validation of several candidate miRNAs. Of 277 miRNAs, five (let-7b, miR-223, miR-25, miR-92a, and miR-423a) were selected as candidates. The expression levels of four miRNAs (let-7b, miR-223, miR-25, miR-92a) were significantly reduced in the lymphoma group, whereas miR-423a levels were significantly increased compared to the controls. When the lymphoma cases were categorized into high- or low-grade as well as into their anatomic form, miR-25 levels were lower in the serum samples from the lymphoma group compared to those from the control group. Although the biological function of serum miRNAs still remains unclear, determining their functional roles in serum and tissues will contribute not only to the identification of potential biomarkers but also to the elucidation of the pathogenesis of canine lymphoma.

    DOI: 10.1016/j.tvjl.2015.04.029

    PubMed

  • Mochizuki H, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H .  Demonstration of the cell clonality in canine hematopoietic tumors by X-chromosome inactivation pattern analysis. .  Vet Pathol52 ( 1 ) 61 - 69   2015年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    X-chromosome inactivation pattern (XCIP) analysis has been widely used to assess cell clonality in various types of human neoplasms. In this study, a polymerase chain reaction-based canine XCIP analysis of the androgen receptor (AR) gene was applied for the assessment of cell clonality in canine hematopoietic tumors. This XCIP analysis is based on the polymorphic CAG repeats in the AR gene and the difference of methylation status between active and inactive X chromosomes. We first examined the polymorphisms of 2 CAG tandem repeats in the AR gene in 52 male and 150 female dogs of various breeds. The 2 polymorphic CAG repeats contained 9 to 12 and 10 to 14 CAGs in the first and second CAG repeats, respectively. Of the 150 female dogs, 74 (49.3%) were heterozygous for the first and/or second polymorphic CAG tandem repeats, indicating the utility of XCIP analysis in these dogs. Canine XCIP analysis was then applied to clinical samples from female dogs with canine high-grade lymphoma, chronic myelogenous leukemia, acute myelogenous leukemia, and benign lymph node hyperplasia. Of 10 lymphoma cell samples, 9 (90%) showed skewed XCIPs, indicating their clonal origins, whereas all the nonneoplastic lymph node samples showed balanced XCIPs. Moreover, bone marrow specimen from a dog with acute myelogenous leukemia and peripheral leukocyte specimens from 2 dogs with chronic myelogenous leukemia showed skewed XCIPs. XCIP analysis was successfully employed to demonstrate the cell clonality of canine hematopoietic tumors in this study and will be applicable to evaluate the clonality in various proliferative disorders in dogs.

    DOI: 10.1177/0300985814528217

    PubMed

  • Sato Masahiko, Mochizuki Hiroyuki, Goto-Koshino Yuko, Fujiwara-Igarashi Aki, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Hypermethylation of the death-associated protein kinase CpG island in canine B-cell lymphoid tumors. .  Vet Immunol Immunopathol161 ( 3-4 ) 222 - 231   2014年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Death-associated protein kinase (DAPK) is a 160-kD serine/threonine kinase known as a key molecule in interferon-gamma (IFN-gamma)-induced apoptosis and tumor suppression. Hypermethylation of the CpG island in DAPK inactivates the gene in a variety of human malignancies. This study aimed to detect the inactivation of DAPK in canine lymphoid tumor cells. The sequence of canine DAPK cDNA was obtained from normal dog peripheral blood mononuclear cells after reverse transcription polymerase chain reaction (RT-PCR). By rapid amplification of 5'-cDNA ends, the transcription initiation site of the DAPK gene was identified. The CpG island located upstream of the translation initiation site was identified by using a search algorithm. The methylation status of the CpG island was examined using bisulfite sequence analysis and methylation-specific PCR (MSP). The inactivation of DAPK gene was examined in 3 canine lymphoid tumor cell lines, GL-1 (B-cell leukemia), CLBL-1 (B-cell lymphoma), and CL-1 (T-cell lymphoma). DAPK mRNA expression was measured by real-time RT-PCR. IFN-gamma-induced apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. The influence of demethylation was examined with 5-aza-2'-deoxycytidine (5-aza-dC). The methylation status in 14 dogs with various lymphoid tumors was screened by MSP. A 1926-bp CpG island containing 280 CpG repeats was identified upstream of the translation start site of canine DAPK. Bisulfate sequence analysis and MSP revealed hypermethylation of the CpG island in GL-1 cells, but not in CLBL-1 or CL-1 cells. The amount of DAPK mRNA was significantly smaller in GL-1 cells than CLBL-1 and CL-1 cells. IFN-gamma-induced apoptosis was detected in CLBL-1 and CL-1 cells but not in GL-1 cells. Treatment with 5-aza-dC significantly increased the amount of DAPK mRNA and IFN-gamma-induced apoptosis in GL-1 cells. These results revealed the inactivation of DAPK through methylation of its CpG island in GL-1 cells. MSP showed hypermethylation of the DAPK CpG island in 5 of 8 primary B-cell lymphoma samples, but not in any of the 6 primary T-cell lymphoid tumor samples obtained from canine patients. DAPK was inactivated through hypermethylation of its CpG island in canine B-cell lymphoid tumor cells. This study will lead to the use of canine B-cell lymphoid tumors as an animal model to evaluate the efficacy of demethylating agents.

    DOI: 10.1016/j.vetimm.2014.08.011

    PubMed

  • Mochizuki Hiroyuki, Seki Takahiro, Nakahara Yoshitaka, Tomita Akitada, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Chronic myelogenous leukaemia with persistent neutrophilia, eosinophilia and basophilia in a cat. .  J Feline Med Surg16 ( 6 ) 517 - 521   2014年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Chronic myelogenous leukaemia was diagnosed in a 7-year-old male neutered domestic shorthair cat. Leukocytosis (74,900/microl)--mature neutrophilia, eosinophilia and basophilia--was observed. Bone marrow aspiration revealed hypercellularity with proliferation of cells of myeloid lineage. An underlying condition leading to leukocytosis was not identified. The severe leukocytosis did not respond to antibiotic therapy. Based on these findings, chronic myelogenous leukaemia was diagnosed. Because of the absence of clinical signs, the cat was monitored without treatment until 7 months after diagnosis, when it developed pruritic skin lesions. Pruritus was controlled with oral prednisolone. Forty-two months after diagnosis, the cat developed nasal lymphoma, which was treated with radiation therapy, resulting in complete remission. The cat was still in good physical condition 63 months after diagnosis, despite the persistence of marked neutrophilia, eosinophilia and basophilia.

    DOI: 10.1177/1098612X13505576

    PubMed

  • Takahashi Masashi, Goto-Koshino Yuko, Fukushima Kenjiro, Kanemoto Hideyuki, Nakashima Ko, Fujino Yasuhito, Ohno Koichi, Endo Yasuyuki, Tsujimoto Hajime .  Phase I dose-escalation study of nimustine in tumor-bearing dogs. .  J Vet Med Sci76 ( 6 ) 895 - 899   2014年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Nimustine (ACNU) is an alkylating agent of the nitrosourea and can be an antineoplastic agent in dogs. But, there has been no report on its dose-limiting toxicity (DLT) in dogs. This study was a phase I dose-escalation clinical trial to determine the maximum tolerated dose (MTD) and DLT of ACNU in tumor-bearing dogs. The starting dosage was 25 mg/m(2), and subsequent dosages were administered in increments of 5 mg/m(2) in cohort of 3 dogs. Eight dogs were included, the MTD was determined to be 25 mg/m(2), DLT was neutropenia, and the optimal interval was considered to be 21 days. The data herein provide a basis for the subsequent phase II trial of ACNU in dogs.

    PubMed

  • Sato Hirofumi, Fujino Yasuhito, Chino Junko, Takahashi Masashi, Fukushima Kenjiro, Goto-Koshino Yuko, Uchida Kazuyuki, Ohno Koichi, Tsujimoto Hajime .  Prognostic analyses on anatomical and morphological classification of feline lymphoma. .  J Vet Med Sci76 ( 6 ) 807 - 811   2014年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The present study was carried out to analyze the prognosis of 163 cats with lymphoma classified anatomically and cytomorphologically. Anatomically, alimentary lymphoma was the most common form and showed significantly shorter survival than mediastinal and nasal lymphomas in cats. Cytomorphologically, there was no predominant subtype in feline lymphomas. Immunoblastic type (18%), centroblastic type (16%), globule leukocyte type (15%), lymphocytic type (12%), lymphoblastic type (12%), pleomorphic medium and large cell type (10%) and anaplastic large cell type (7%) were relatively common subtypes. Most of the cats with globule leukocyte lymphoma had the alimentary form. Comparing median survival time among classifications, cats with globule leukocyte lymphoma showed significantly shorter survival than those with high-grade and other low-grade lymphomas. Furthermore, cats with high-grade lymphomas showed significantly shorter survival than cats with other low-grade lymphomas. The present study indicated the clinical significance of anatomical and cytomorphological evaluation in feline lymphomas.

    PubMed

  • Takahashi Masashi, Tomiyasu Hirotaka, Hotta Eri, Asada Hajime, Fukushima Kenjiro, Kanemoto Hideyuki, Fujino Yasuhito, Ohno Koichi, Uchida Kazuyuki, Nakayama Hiroyuki, Tsujimoto Hajime .  Clinical characteristics and prognostic factors in dogs with histiocytic sarcomas in Japan. .  J Vet Med Sci76 ( 5 ) 661 - 666   2014年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Canine histiocytic sarcoma (HS) is a rare neoplasm that originates from dendritic cells or macrophages, and there have been a number of cases experienced in Japan. To identify the characteristics and prognostic variables that determine outcome in dogs with HS in Japan, medical records of 73 dogs with HS were retrospectively analyzed. Signalment, clinical signs, complete blood count (CBC), blood chemistry profiles, treatment, response to treatment and overall survival (OS) were analyzed. Diagnosis of HS was determined histologically in 44 cases and cytologically in 29 cases. The most frequently diagnosed breeds were Flat-Coated Retrievers (n=16, odds ratio [OR] 62.0), Pembroke Welsh corgis (n=15, OR 9.7) and Bernese Mountain dogs (n=14, OR 45.0). Median survival time for all dogs in this study was 43 days. In the dogs that received no treatment or only symptomatic treatment, the median OS was 12 days (range 2-254 days) compared with that of dogs that received surgical treatment and/or chemotherapy (85 days, range 4-360 days). Univariate analysis identified anemia, thrombocytopenia, hypoalbuminemia, hypoproteinemia and not receiving antitumor treatment (chemotherapy and/or surgery) as factors significantly associated with shorter OS. Multivariate analysis confirmed that platelet counts, localized/disseminated lesional pattern and whether the dog received antitumor treatment were significantly predictive of survival.

    PubMed

  • Fukushima K, Kanemoto H, Ohno K, Takahashi M, Fujiwara R, Nishimura R, Tsujimoto H .  Computed tomographic morphology and clinical features of extrahepatic portosystemic shunts in 172 dogs in Japan. .  Vet J199 ( 3 ) 376 - 381   2014年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Canine extrahepatic congenital portosystemic shunts (EH-cPSS) are classified into several anatomical types, depending on the origin and termination of the shunt vessel. The aim of this retrospective study was to determine the proportion and clinical features of each anatomical shunt type in a population of dogs presented to a veterinary teaching hospital in Japan. Dogs diagnosed with EH-cPSS using computed tomographic (CT) portography were included (n=172) and shunts were classified based on previous reports. Clinical data were collected from case records and analysed statistically. The most common anatomical type was the spleno-phrenic shunt (n=64), followed by the spleno-azygos (n=38), right gastric-caval (n=29), spleno-caval (n=21), right gastric-caval with caudal loop (n=9), right gastric-phrenic (n=6), colono-caval (n=3), spleno-phrenic and azygos (n=1), and porto-caval (n=1) shunts. Spleno-phrenic and spleno-azygos shunts were diagnosed more frequently in older dogs than right gastric-caval and spleno-caval shunts (P<0.05). The portal vein/aortic (PV/Ao) ratio was significantly larger in dogs with spleno-phrenic shunts than in dogs with spleno-azygos, right gastric-caval or spleno-caval shunts (P<0.05). The PV/Ao ratio was significantly larger in dogs with spleno-azygos shunts than in dogs with right gastric-caval shunts. Dogs with spleno-phrenic shunts had significantly lower serum alkaline phosphatase activities than those with right gastric-caval or spleno-caval shunts. Dogs with spleno-phrenic shunts had significantly lower fasting ammonia concentrations than those with spleno-caval shunts.

    DOI: 10.1016/j.tvjl.2013.11.013

    PubMed

  • Fujiwara-Igarashi Aki, Goto-Koshino Yuko, Sato Masahiko, Maeda Shingo, Igarashi Hirotaka, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Prognostic significance of the expression levels of the p16, p15, and p14 genes in dogs with high-grade lymphoma. .  Vet J199 ( 2 ) 236 - 244   2014年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The prognostic significance of the inactivation of the p16, p15, and p14 genes has been reported in lymphoid malignancies in humans. To evaluate the relationship between inactivation of the p16, p15, and p14 genes and prognosis in canine high-grade lymphoma, primary tumor cell samples obtained from 71 dogs with high-grade lymphoma were examined for the expression levels of these genes. Quantitative and conventional reverse transcriptase-polymerase chain reaction (RT-PCR) analyses were used to measure the amounts of p16, p15, and p14 mRNAs. The methylation status of the CpG island of the p16 gene was evaluated using methylation-specific PCR. Overall survival (OS) was compared using the Kaplan-Meier method. The log-rank test and Cox proportional hazard model were used to evaluate factors that influenced OS. Of 62 dogs examined, p16, p15, and p14 mRNA levels were found to be undetectable in 21, 18, and 10 dogs, respectively. In 20/68 dogs analyzed, the CpG island of the p16 gene was shown to be methylated. The prognostic significance of inactivation of the p16, p15, and p14 genes as well as various conventional factors obtained from medical records was examined. p16 expression status and anatomic form/immunophenotype were found to correlate with OS in the dogs with high-grade lymphoma. p16 mRNA level over its cut-off value correlated with a poor prognosis; however, the expression levels of p15 and p14 mRNAs and p16 methylation status did not influence the prognosis in dogs with high-grade lymphoma.

    DOI: 10.1016/j.tvjl.2013.11.004

    PubMed

  • Fujino Yasuhito, Nakamura Yoichi, Matsumoto Hideaki, Fukushima Kenjiro, Takahashi Masashi, Ohno Koichi, Tsujimoto Hajime .  Development and evaluation of a novel in-clinic automated hematology analyzer, ProCyte Dx, for canine erythrocyte indices, leukogram, platelet counts and reticulocyte counts. .  J Vet Med Sci75 ( 11 ) 1519 - 1524   2013年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A novel hematology analyzer for small animal medicine, ProCyte Dx, was developed from combination of the fluorescence laser flow cytometry and laminar flow impedance technologies, and its accuracy was evaluated by comparing with the conventional impedance-based hematology analyzer, pocH-100iV Diff, or microscopic manual cell counting methods with staining blood smears in the canine blood. Blood samples of 59 dogs were hematologically analyzed and compared by Pearson's correlation coefficients. Analyses between the two analyzers showed excellent correlation in RBC (r=0.998), HGB (r=0.999), HCT (r=0.998), MCV (r=0.994), MCH (r=0.974), MCHC (r=0.906), WBC (r=0.998) and PLT (r=0.993). Analyses between ProCyte Dx and microscopic manual counting results showed excellent correlation in neutrophils (r=0.920), lymphocytes (r=0.913) and reticulocyte percentages (r=0.924), good correlation in eosinophils (r=0.815) and reticulocyte numbers (r=0.850) and fair correlation in monocytes (r=0.770). The present study indicates that ProCyte Dx is acceptably accurate and can be a powerful tool for canine clinical medicine.

    PubMed

  • Takeuchi Yoshinori, Fujino Yasuhito, Watanabe Manabu, Takahashi Masashi, Nakagawa Takayuki, Takeuchi Ayano, Bonkobara Makoto, Kobayashi Tetsuya, Ohno Koichi, Uchida Kazuyuki, Asano Kazushi, Nishimura Ryohei, Nakayama Hiroyuki, Sugano Sumio, Ohashi Yasuo, Tsujimoto Hajime .  Validation of the prognostic value of histopathological grading or c-kit mutation in canine cutaneous mast cell tumours: a retrospective cohort study. .  Vet J196 ( 3 ) 492 - 498   2013年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The objective of this retrospective cohort study was to validate the prognostic value of histological grading of canine cutaneous mast cell tumours (MCTs) according to the Patnaik (grades I-III) and Kiupel (low, high) grading systems, and to confirm the prognostic significance of internal tandem duplications (ITDs) within exon 11 of the c-kit gene (ITD-Exon11). The baseline characteristics and outcome data from 47 dogs diagnosed with cutaneous MCTs were collected and reviewed. Tumours were graded according to both grading systems and the nucleotide sequence of c-kit was evaluated. Results were analyzed to evaluate predictive factors for overall survival (OS) and progression-free survival (PFS). Log-rank tests indicated that dogs with Patnaik grade III MCTs had significantly reduced OS and PFS compared to those with either grade I or II tumours. However, no significant difference in OS or PFS was observed between grade I and II tumours. The dogs with Kiupel high-grade MCTs had significantly shorter OS and PFS than dogs with low-grade MCTs. The presence of ITD-Exon11 was significantly associated with shorter PFS. The result of Cox regression analysis showed that the Kiupel grading system for OS and PFS, and lymph node metastasis for OS, independently predicted prognosis. Kappa statistics confirmed a significantly higher inter-observer consistency for the Kiupel compared to the Patnaik grading system. These findings demonstrate that the Kiupel grading system is a useful prognostic tool for canine cutaneous MCTs in predicting OS and PFS, while the occurrence of ITD-Exon11 appeared to be a useful predictor for PFS.

    DOI: 10.1016/j.tvjl.2012.11.018

    PubMed

  • Tamamoto Takashi, Ohno Koichi, Takahashi Masashi, Nakashima Ko, Fujino Yasuhito, Tsujimoto Hajime .  Serum amyloid A as a prognostic marker in cats with various diseases. .  J Vet Diagn Invest25 ( 3 ) 428 - 432   2013年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Serum amyloid A (SAA) is reported not only as a marker for the presence of inflammation but also as a prognostic indicator in human beings. In cats, however, there is no report on the association between SAA concentration and prognosis. The objective of the current study was to evaluate SAA concentration as a prognostic marker in diseased cats. A total of 175 cats with neoplastic diseases, inflammatory diseases, and other diseases were retrospectively recruited, and the medical records of these cats, including follow-up data on mortality, were reviewed. Cats were divided into 2 groups according to SAA concentration, and differences in survival between each group were assessed. Median survival time of cats in the elevated SAA (>0.82 mg/l) group was significantly shorter than that in the nonelevated SAA (</=0.82 mg/l) group (P < 0.001). Furthermore, by multivariate analysis, SAA concentration was shown as a significant and independent prognostic marker in cats with various diseases (P = 0.015). Serum amyloid A concentration in diseased cats is a useful predictive indicator of prognosis regardless of diagnosis.

    DOI: 10.1177/1040638713486112

    PubMed

  • Maeda Shingo, Fujino Yasuhito, Tamamoto Chie, Suzuki Satoshi, Fujita Atsushi, Takahashi Masashi, Ohno Koichi, Uchida Kazuyuki, Tsujimoto Hajime .  Lymphangiomatosis of the systemic skin in an old dog. .  J Vet Med Sci75 ( 2 ) 187 - 190   2013年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A 13-year-old, neutered male miniature dachshund was presented with a one-month history of bilateral symmetrical swelling in the pinnae and carpal, cubital and tarsal joints, and swelling in the tail. The lesions were histopathologically characterized by multiple dilated lymphatic vessels lined by a single attenuated layer of endothelial cells. The subcutis was predominantly involved. A number of spindle-shaped cells lining the irregular vessels were observed. Morphological atypia was not evident in these cells. Immunohistochemical analyses revealed that the proliferating endothelial cells were positive for factor VIII-related antigen and CD31. Based on the clinical presentation and histopathological features, the dog was diagnosed with lymphangiomatosis. Treatment with anti-inflammatory prednisolone improved the symptoms.

    PubMed

  • Mochizuki H, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H .  X-chromosome inactivation pattern analysis for the assessment of cell clonality in cats. .  Vet Pathol49 ( 6 ) 963 - 970   2012年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    X-chromosome inactivation pattern (XCIP) analysis has been widely used to assess cell clonality in various types of neoplasms in humans. In the present study, a polymerase chain reaction-based feline XCIP analysis using the feline androgen receptor gene was developed. To construct the system of the analysis, polymorphism in CAG tandem repeats within the feline androgen receptor gene was explored using somatic DNAs from 50 male and 103 female cats. CAG tandem repeats in exon 1 of the feline androgen receptor gene were found to be polymorphic, containing 15 to 22 CAG repeats. Of the 103 female cats, 70 (68%) were heterozygous for the number of CAG repeats, indicating the possible usefulness of XCIP analysis in cats. Application of the feline XCIP analysis to 3 feline mammary gland adenocarcinoma cell lines revealed distinctly skewed XCIPs in these cell lines, indicating their clonal origins. Twelve (80%) of the 15 primary tissue/cell samples obtained from cats with various neoplastic diseases showed skewed XCIPs. Moreover, bone marrow samples from 3 cats with myelodysplastic syndrome were also found to have skewed XCIPs. The polymerase chain reaction-based XCIP analysis developed in this study can provide information on cell clonality in female cats, potentially facilitating the differential diagnosis of various disorders in cats.

    DOI: 10.1177/0300985812439216

    PubMed

  • Takeuchi Yoshinori, Takahashi Masashi, Tsuboi Masaya, Fujino Yasuhito, Uchida Kazuyuki, Ohno Koichi, Nakayama Hiroyuki, Tsujimoto Hajime .  Intestinal T-cell lymphoma with severe hypereosinophilic syndrome in a cat. .  J Vet Med Sci74 ( 8 ) 1057 - 1062   2012年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A Japanese domestic long-hair cat of about 8 years of age was presented with vomiting and hematochezia and was found to have significant hypereosinophilia. Bone marrow aspiration revealed moderate increases of eosinophilic lineages. Histopathological examination revealed mild eosinophilic and epitheliotropic T-lymphocytic infiltrations in the duodenum. Although the cat remained asymptomatic with only prednisolone administration, the cat presented with hematemesis, weight loss, and severe anorexia 512 days after the initial presentation. Subsequently, gastrointestinal perforation developed, and the cat died on Day 536. Histopathological examination of autopsy specimens revealed mixed cellular infiltration including eosinophils and neoplastic lymphocytes in the intestinal lymph nodes, intestine, liver, spleen, and pancreas. Immunohistochemical examination supports a diagnosis of intestinal T-cell lymphoma with severe hypereosinophilic syndrome.

    PubMed

  • Tei Meina, Uchida Kazuyuki, Chambers James K, Harada Hiromi, Takahashi Masashi, Nishimura Ryohei, Watanabe Manabu, Nakayama Hiroyuki .  Mammary lipid-rich carcinoma with extensive amyloid deposition in a dog. .  J Vet Med Sci74 ( 6 ) 809 - 811   2012年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A 6-year-old female Labrador retriever presented with a mass in the right mammary gland, and swollen right inguinal and axillary lymph nodes. Fine needle aspiration biopsy suggested a malignant lipid-producing tumor, such as liposarcoma. Histopathologically, the neoplasms were solid, lobulated nests of atypical epithelial cells with a large amount of extracellular deposits of amyloid in both mammary gland and lymph nodes. The proliferating cells contained large cytoplasmic vacuoles, positive for oil red-O. These cells were immunopositive for cytokeratin (AE1/AE3) and beta-casein and negative for SMA. The amyloid deposits were immunopositive for beta-casein. These findings suggested that the proliferating cells secreted beta-casein forming amyloid deposits. This is the first report of mammary lipid-rich carcinoma with extensive amyloid deposition derived from beta-casein.

    PubMed

  • Fukushima Kenjiro, Kanemoto Hideyuki, Ohno Koichi, Takahashi Masashi, Nakashima Ko, Fujino Yasuhito, Uchida Kazuyuki, Fujiwara Reina, Nishimura Ryohei, Tsujimoto Hajime .  CT characteristics of primary hepatic mass lesions in dogs. .  Vet Radiol Ultrasound53 ( 3 ) 252 - 257   2012年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Little information is available on the relationship between computed tomography (CT) imaging findings and the pathologic diagnosis of canine hepatic tumors. Our purpose was to clarify the characteristic features of CT findings in liver tumors in dogs. Data from 33 dogs with either a hepatocellular carcinoma, n = 14, hepatocellular adenoma, n = 14, or nodular hyperplasia, n = 5 were summarized from medical records. CT features for each histologic diagnosis were characterized and analyzed statistically. Common findings in hepatocellular carcinoma included central (79%, P = 0.0030) and marginal enhancement (93%, P = 0.00043) in the arterial phase, cyst-like lesions (93%), capsule formation (93%), and hypoattenuation in the portal (86%), and equilibrium phases (93%). Hepatic adenoma was characterized by a characteristic diffuse enhancement pattern during the arterial phase (57%, P = 0.013), which was also found in nodular hyperplasia (60%), but never in hepatocellular carcinoma. Nodular hyperplasia was less likely to have a capsule structure (20%, P = 0.0087). Mass size was significantly smaller in nodular hyperplasia than in hepatocellular carcinoma and hepatic adenoma (P = 0.0033 and 0.038, respectively). Hyperattenuation in the arterial and the portal phase i.e. contrast retention, was more frequent in hepatic adenoma than in the other groups (P = 0.037 and 0.037, respectively). Nodular hyperplasia was more frequently isoattenuating in the equilibrium phase (P = 0.043).

    DOI: 10.1111/j.1740-8261.2011.01917.x

    PubMed

  • Mochizuki Hiroyuki, Nakamura Kenji, Sato Hirofumi, Goto-Koshino Yuko, Sato Masahiko, Takahashi Masashi, Fukushima Kenjiro, Nakashima Ko, Fujino Yasuhito, Ohno Koichi, Uchida Kazuyuki, Nakayama Hiroyuki, Tsujimoto Hajime .  GeneScan analysis to detect clonality of T-cell receptor gamma gene rearrangement in feline lymphoid neoplasms. .  Vet Immunol Immunopathol145 ( 1-2 ) 402 - 409   2012年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. Polymerase chain reaction (PCR) amplification of the complementarity-determining region (CDR) 3 of the T-cell receptor (TCR) gamma gene can be used to assess clonality of T-cell populations as a supportive diagnostic tool for T-cell neoplasms. Because the length variation in the TCRgamma CDR3 is relatively small, false positive results may occur in non-neoplastic T-cell populations in the absence of high-resolution analytical methods for PCR products. In the present study, a PCR assay system was developed to detect clonal TCRgamma gene rearrangement in feline lymphoid cells using GeneScan analysis. Thirty T-cell neoplasms, 27 B-cell neoplasms, and 34 non-neoplastic tissues were subjected to the newly developed TCRgamma gene rearrangement analysis. Clonal TCRgamma gene rearrangement was detected in 26 of 30 (87%) T-cell neoplasms, 2 of 27 (7%) B-cell neoplasms, and 1 of 34 (3%) non-neoplastic tissues. To compare GeneScan analysis with conventional PAGE and heteroduplex analysis, 20 clonal and 20 polyclonal samples were subjected to both analyses. Most of the results were concordant between the 2 analyses; however, several clonal peaks (bands) appeared as a single band when analyzed via conventional PAGE with heteroduplex analysis in 4 of the 20 (20%) clonal samples as a result of the difference in resolution. The PCR assay system to detect clonal TCRgamma gene rearrangement in feline lymphoid cells, using GeneScan analysis, would be a useful molecular diagnostic tool for feline T-cell neoplasms, with high fidelity.

    DOI: 10.1016/j.vetimm.2011.12.015

    PubMed

  • Sato Masahiko, Kanemoto Hideyuki, Kagawa Yumiko, Kobayashi Tetsuya, Goto-Koshino Yuko, Mochizuki Hiroyuki, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Evaluation of the prognostic significance of BCL6 gene expression in canine high-grade B-cell lymphoma. .  Vet J191 ( 1 ) 108 - 114   2012年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The clinical usefulness of BCL6 gene expression was evaluated as a prognostic indicator in dogs with high-grade B-cell lymphoma. Forty-four dogs were diagnosed with centroblastic or B-cell immunoblastic type lymphoma according to the updated Kiel classification. BCL6 mRNA expression was measured by real-time PCR and its relationship with prognosis was analyzed. Progression-free and overall survival was not significantly different between the high BCL6 expression group (higher than the median) and the low BCL6 expression group (lower than the median) (P=0.99 and P=0.61, respectively). No correlation between BCL6 and prognosis was observed in this study, which is inconsistent with findings reported for human diffuse large B-cell lymphoma. BCL6 protein expression was not detected in the 11 dogs evaluated by immunohistochemistry. Furthermore, BCL6 protein expression was assessed in 13 archived paraffin-embedded high-grade canine lymphoma tissues and all were also negative. The results suggest that most canine high-grade B-cell lymphomas correspond to human diffuse large B-cell lymphoma with no immunohistochemical expression of BCL6.

    DOI: 10.1016/j.tvjl.2010.12.006

    PubMed

  • Tsukagoshi Taro, Ohno Koichi, Tsukamoto Atsushi, Fukushima Kenjiro, Takahashi Masashi, Nakashima Ko, Fujino Yasuhito, Tsujimoto Hajime .  Decreased gallbladder emptying in dogs with biliary sludge or gallbladder mucocele. .  Vet Radiol Ultrasound53 ( 1 ) 84 - 91   2012年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Biliary sludge in dogs is dismissed commonly as an incidental finding. On the other hand, gallbladder mucocele is reported increasingly in dogs and can lead to biliary obstruction or gallbladder rupture. Cholestasis is suspected to play a role in development of sludge and mucoceles, though there are no data in dogs to support this. We investigated gallbladder emptying, a key factor in biliary flow, in dogs with mobile sludge, immobile sludge, or gallbladder mucocele and in healthy controls. Gallbladder ejection fraction estimated by ultrasonography was used as the index of gallbladder emptying. The ejection fraction at 60 min after eating was significantly decreased in all three abnormal groups. Moreover, all dogs with sludge or a mucocele had gallbladder distension. These changes were the greatest in the mucocele group. Thus, biliary stasis occurs not only in dogs with gallbladder mucocele but also in dogs with biliary sludge. Cholestasis may play a role in the pathogenesis or progression of these diseases in dogs.

    DOI: 10.1111/j.1740-8261.2011.01868.x

    PubMed

  • Ohmi Aki, Tsukamoto Atsushi, Ohno Koichi, Uchida Kazuyuki, Nishimura Ryohei, Fukushima Kenjiro, Takahashi Masashi, Nakashima Ko, Fujino Yasuhito, Tsujimoto Hajime .  A retrospective study of inflammatory colorectal polyps in miniature dachshunds. .  J Vet Med Sci74 ( 1 ) 59 - 64   2012年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Medical records of dogs with colorectal polyps were retrospectively reviewed, and clinical presentation of inflammatory colorectal polyps in miniature dachshunds was evaluated. Of 33 dogs found to have colorectal polyps, miniature dachshunds were markedly over-represented with 16 dogs (48%), of which 12 (75%) were found to have inflammatory polyps. Multiple polyps localized between the rectum and the descending colon was the most common finding in miniature dachshunds with inflammatory polyps. Twenty dogs (80%) out of 25 miniature dachshunds with inflammatory colorectal polyps responded to immunosuppressive therapy using prednisolone and cyclosporine. The results of this study indicate that miniature dachshunds are predisposed to develop inflammatory colorectal multiple polyps, for which immunosuppressive therapy may be a treatment option.

    PubMed

  • Mochizuki Hiroyuki, Nakamura Kenji, Sato Hirofumi, Goto-Koshino Yuko, Sato Masahiko, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Uchida Kazuyuki, Nakayama Hiroyuki, Tsujimoto Hajime .  Multiplex PCR and Genescan analysis to detect immunoglobulin heavy chain gene rearrangement in feline B-cell neoplasms. .  Vet Immunol Immunopathol143 ( 1-2 ) 38 - 45   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.

    DOI: 10.1016/j.vetimm.2011.05.030

    PubMed

  • Ishida Asami, Ohno Koichi, Fukushima Kenjiro, Nakashima Ko, Takahashi Masashi, Goto-Koshino Yuko, Fujino Yasuhito, Tsujimoto Hajime .  Plasma high-mobility group box 1 (HMGB1) in dogs with various diseases: comparison with C-reactive protein. .  J Vet Med Sci73 ( 9 ) 1127 - 1132   2011年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    High-mobility group box 1 (HMGB1), a nonhistone chromosomal protein, has recently been suggested as a late mediator of the inflammatory cascade. Blood HMGB1 levels are increased in a number of human diseases, and HMGB1 has been suggested to be a useful marker for disease severity and prognosis. The objective of this study was to assess the clinical usefulness of HMGB1 in dogs. Plasma HMGB1 levels, as well as C-reactive protein (CRP), a typical canine inflammatory marker, were measured in dogs with various diseases, especially systemic inflammatory response syndrome (SIRS), and dogs that had undergone surgery. HMGB1 gradually increased and attained a maximum level 72 hr after surgery, whereas CRP increased rapidly, peaking at 24 hr. Although both HMGB1 and CRP levels were significantly increased in dogs with various diseases compared with the control dogs, no correlation was found between the HMGB1 and CRP values. HMGB1 levels in the SIRS group were significantly elevated compared with those in the non-SIRS group. However, the increase in HMGB1 levels above the reference range was not indicative of SIRS. Instead, the presence of increased HMGB1 and CRP levels above the reference ranges significantly affects the poor outcome of SIRS. The present study indicates that HMGB1 is a novel canine inflammatory marker and is distinct from CRP. However, the additional clinical value of HMGB1 measurement remains unclear, and further studies are warranted.

    PubMed

  • Tomiyasu H, Fujino Y, Takahashi M, Ohno K, Tsujimoto H .  Spontaneous acute erythroblastic leukaemia (AML-M6Er) in a dog. .  J Small Anim Pract52 ( 8 ) 445 - 447   2011年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A five-year-old golden retriever was presented with anaemia, thrombocytopenia, anorexia and lethargy. Peripheral blood cytology showed abnormal cells similar to proerythroblasts with multiple nucleoli and strongly basophilic cytoplasm. Bone marrow cytopathology revealed that the blast cells accounted for more than 80% of all nucleated cells (ANC). These blast cells were confirmed as erythroblastic cells by cytochemistry, polymerase chain reaction for genetic clonality assessment of IgH and TCRgamma, flow cytometry, and transmission electron microscopy. Based on these observations, the dog was diagnosed with acute erythroblastic leukaemia (AML-M6Er). Chemotherapy with cytarabine commenced on day 7 after initial presentation, but the dog died 2 days later. This is the first report of spontaneous AML-M6Er in a dog.

    DOI: 10.1111/j.1748-5827.2011.01096.x

    PubMed

  • Kanemoto H, Ohno K, Sakai M, Nakashima K, Takahashi M, Fujino Y, Tsujimoto H .  Expression of fibrosis-related genes in canine chronic hepatitis. .  Vet Pathol48 ( 4 ) 839 - 845   2011年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Molecular regulation of fibrosis in chronic canine hepatitis is poorly understood. The authors employed quantitative polymerase chain reaction (PCR) to determine the expression levels of genes reported to be related to fibrosis in other species (human, mouse, and rat) and to elucidate the relationship of these genes with the degree of fibrosis and the presence or absence of ascites and/or jaundice in dogs with hepatitis. Nine fibrosis-related genes were assayed: PDGFB, PDGFD, MMP2, TIMP1, THBS1, COL1A1, COL3A1, TGFB1, and TGFB2. Liver samples of 15 dogs with chronic hepatitis and 4 healthy control dogs were obtained via laparoscopic biopsy and subjected to histologic and quantitative PCR analyses. The expression of all 9 genes showed significant positive correlation (P<.01, r>.70) with the degree of fibrosis. Furthermore, the expression levels of all genes except TGFB1 were significantly higher (P<.05) in dogs with hepatic failure-related symptoms (ascites/jaundice). Results suggest that these 9 genes are integral to the development of fibrosis in canine chronic hepatitis.

    DOI: 10.1177/0300985810388523

    PubMed

  • Mochizuki Hiroyuki, Takahashi Masashi, Nishigaki Kazuo, Ide Tetsuya, Goto-Koshino Yuko, Watanabe Shinya, Sato Hirofumi, Sato Masahiko, Kotera Yukiko, Fujino Yasuhito, Ohno Koichi, Uchida Kazuyuki, Tsujimoto Hajime .  Establishment of a novel feline leukemia virus (FeLV)-negative B-cell cell line from a cat with B-cell lymphoma. .  Vet Immunol Immunopathol140 ( 3-4 ) 307 - 311   2011年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We established a novel feline B-cell line, MS4, from the neoplastic pleural effusion of a cat with cutaneous B-cell lymphoma. Immunophenotype staining of the MS4 cells was positive for CD20, CD79alpha, and IgA and negative for CD3, CD4, CD5, CD8alpha, CD18, CD21, CD22, IgM, IgG, Ig light chain, and MHC class II. PCR analysis for immunoglobulin heavy chain gene rearrangements revealed a monoclonal rearrangement, whereas no clonal rearrangement of the T-cell receptor gamma gene was detected. Southern blotting with an exogenous feline leukemia virus (FeLV) U3 probe revealed no integration of exogenous FeLV provirus. The MS4 cell line is the first FeLV-negative feline B-cell lymphoma cell line, and may be used to investigate the pathogenesis of spontaneously occurring feline lymphoma and the development of new therapies.

    DOI: 10.1016/j.vetimm.2010.12.010

    PubMed

  • Sato M, Yamazaki J, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H .  Increase in minimal residual disease in peripheral blood before clinical relapse in dogs with lymphoma that achieved complete remission after chemotherapy. .  J Vet Intern Med25 ( 2 ) 292 - 296   2011年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: We developed previously a minimal residual disease (MRD) monitoring system in dogs with lymphoma by exploring a highly sensitive real-time PCR system. OBJECTIVES: To identify the change in MRD before clinical relapse in dogs with lymphoma that achieved complete remission after chemotherapy. ANIMALS: Twenty dogs with multicentric high-grade B-cell lymphoma. METHODS: MRD levels in peripheral blood mononuclear cells (PBMCs) were measured by real-time PCR amplifying the rearranged immunoglobulin heavy chain gene. MRD measurement and clinical assessment were performed every 2-4 weeks for 28-601 days after completion of chemotherapy. An increase in MRD was defined as an increase by more than 0.5, calculated by log(1)(0) [copy number of MRD per 10(5) PBMCs], based on the uncertainty level observed in a canine lymphoma cell line. RESULTS: During the follow-up period, 15 dogs relapsed in 28-320 days (median, 120 days) after completion of chemotherapy. An increase in MRD was detected 2 weeks or more before relapse in 14 of the 15 dogs, but an increase in MRD before relapse could not be detected in the remaining 1 dog. The time from increased MRD to clinical relapse was 0-63 days (median, 42 days). In contrast, no increase in MRD was detected in 5 dogs that did not experience clinical relapse. CONCLUSION AND CLINICAL IMPORTANCE: An increase in MRD can be detected before clinical relapse in dogs with lymphoma. Application of early reinduction therapy based on an increase in MRD before clinical relapse may improve treatment outcome in canine lymphoma.

    DOI: 10.1111/j.1939-1676.2010.0675.x

    PubMed

  • Sato M, Yamazaki J, Goto-Koshino Y, Takahashi M, Fujino Y, Ohno K, Tsujimoto H .  Evaluation of cytoreductive efficacy of vincristine, cyclophosphamide, and Doxorubicin in dogs with lymphoma by measuring the number of neoplastic lymphoid cells with real-time polymerase chain reaction. .  J Vet Intern Med25 ( 2 ) 285 - 291   2011年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: The cytoreductive efficacy of the individual components of multidrug chemotherapy for canine lymphoma is difficult to evaluate after complete remission. OBJECTIVES: To compare the cytoreductive efficacy of vincristine (VCR), cyclophosphamide (CPA), and doxorubicin (DXR) in dogs that received a 6-month modified version of the University of Wisconsin-Madison chemotherapy protocol (UW-25). ANIMALS: Twenty-nine dogs with high-grade B-cell multicentric lymphoma. METHODS: Rearranged immunoglobulin heavy chain gene fragments from lymphoma cells were amplified by polymerase chain reaction (PCR) and sequenced to prepare clone-specific primers and probes for real-time PCR. The number of lymphoma cells in peripheral blood was measured from diagnosis to week 11 of UW-25. RESULTS: The number of lymphoma cells after the 1st administration of VCR, CPA, and DXR in weeks 1-4 was decreased in 29/29 (100%), 15/29 (51.7%), and 26/27 (96.3%) dogs, respectively. The cytoreductive efficacy of CPA was less than that of VCR and DXR. VCR, CPA, and DXR administered in weeks 6-9 were effective in 5/26 (19.2%), 5/20 (25.0%), and 14/19 (73.7%) dogs, respectively, indicating the sustained cytoreductive efficacy of DXR. CPA nonresponders were heavier and exhibited a shorter 1st remission than CPA responders. CONCLUSION AND CLINICAL IMPORTANCE: When using UW-25 for treatment of canine lymphoma, CPA was found to have less cytoreductive efficacy than VCR and DXR. Real-time PCR-based quantification of tumor cells is an objective marker of the efficacy of chemotherapeutic agents.

    DOI: 10.1111/j.1939-1676.2011.0686.x

    PubMed

  • Ohmi Aki, Ohno Koichi, Uchida Kazuyuki, Nakayama Hiroyuki, Koshino-Goto Yuko, Fukushima Kenjiro, Takahashi Masashi, Nakashima Ko, Fujino Yasuhito, Tsujimoto Hajime .  A retrospective study in 21 Shiba dogs with chronic enteropathy. .  J Vet Med Sci73 ( 1 ) 1 - 5   2011年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We retrospectively studied the clinical and laboratory features and outcomes of chronic enteropathy in Shiba dogs. Among 99 dogs with chronic enteropathy, 21 Shiba dogs (21%) were included in the study (odds ratio, 7.14). No significant differences were seen in signalment, clinical signs, symptoms or laboratory profiles between the Shiba and non-Shiba groups. Severe histopathological lesions in the duodenum were a common finding in the Shiba group. The median overall duration of survival in the Shiba group was 74 days, while that of the dogs in the non-Shiba group could not be determined because more than half of the cases remained alive at the end of this study. The difference between the groups was statistically significant (P<0.0001). The 6-month and 1-year survival rates for the Shiba group were 46% and 31%, respectively. Conversely, the 6-month, 1-year and 3-year survival rates for the non-Shiba group were 83%, 74% and 67%. The results obtained here demonstrated that the Shiba dog is predisposed to chronic enteropathy and shows severe duodenum lesions and poor outcomes, indicating a breed-specific disease.

    PubMed

  • Tomiyasu Hirotaka, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Gastrointestinal and hematologic adverse events after administration of vincristine, cyclophosphamide, and doxorubicin in dogs with lymphoma that underwent a combination multidrug chemotherapy protocol. .  J Vet Med Sci72 ( 11 ) 1391 - 1397   2010年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The present study aimed to objectively evaluate the adverse events after the administration of chemotherapeutic agents used in the University of Wisconsin (UW)-Madison chemotherapy protocol (UW-25) for canine lymphoma, using the Veterinary Co-operative Oncology Group common terminology criteria for adverse events (VCOG-CTCAE). The medical records of 40 dogs with multicentric high-grade lymphoma that underwent UW-25 were reviewed. Gastrointestinal adverse events of grade 2 and above and blood/bone marrow adverse events of all grades were evaluated. Gastrointestinal adverse events occurring at least once during the entire period of UW-25 were observed in 50% (20/40), 17.9% (7/39), and 8.1% (3/37) of the dogs after the administration of vincristine (VCR), cyclophosphamide (CPA), and doxorubicin (DXR), respectively. Blood/bone marrow adverse events occurring at least once during UW-25 were observed in 57.5% (23/40), 41% (16/39), and 8.1% (3/37) of the dogs after the administration of VCR, CPA, and DXR, respectively. The rate of patients that experienced gastrointestinal adverse events was higher after the first administration of VCR than after the first administration of DXR. Findings obtained in this study will be helpful in predicting the adverse events that could occur when dogs with lymphoma are treated with UW-25.

    PubMed

  • Tomiyasu Hirotaka, Fujino Yasuhito, Ugai Jun, Goto-Koshino Yuko, Ide Tetsuya, Takahashi Masashi, Ohno Koichi, Uchida Kazuyuki, Nakayam Hiroyuki, Tsujimoto Hajime .  Eosinophilia and eosinophilic infiltration into splenic B-cell high-grade lymphoma in a dog. .  J Vet Med Sci72 ( 10 ) 1367 - 1370   2010年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A 13-year-old mixed-breed dog showing ascites, anorexia and anemia was found to have leukocytosis with marked eosinophilia, splenomegaly and hepatomegaly. The dog died 4 days after initial presentation and was diagnosed with splenic high-grade B-cell lymphoma at necropsy. Remarkable infiltrations of eosinophils were observed in spleen and liver tissues. The eosinophilia and infiltration of eosinophils into the lesions could have been associated with B-cell lymphoma because causes other than lymphoma were excluded. This is the first report of eosinophilia and eosinophilic infiltrations into neoplastic lesions in a dog with high-grade B-cell lymphoma.

    PubMed

  • Tomiyasu Hirotaka, Goto-Koshino Yuko, Takahashi Masashi, Fujino Yasuhito, Ohno Koichi, Tsujimoto Hajime .  Quantitative analysis of mRNA for 10 different drug resistance factors in dogs with lymphoma. .  J Vet Med Sci72 ( 9 ) 1165 - 1172   2010年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Expression levels of ABCB1, ABCC1, Lung resistance-associated protein (LRP), ABCG2, p53, p21(waf1), Bcl-2, CD40L, glutathione S-transferase alpha (GSTalpha), and O(6)-methylguanine-DNA-methyltransferase (MGMT) genes, and mutation of p53 gene were examined in 23 dogs with multicentric high-grade lymphoma to explore their association with drug resistance of the tumor cells. Dogs were divided into chemotherapy-sensitive (n=13) and -resistant (n=10) groups according to the response to a 6-month modified version of the University of Wisconsin (UW)-Madison chemotherapy protocol (UW-25), and expression levels of these factors and frequency of p53 gene mutation were compared between groups. No significant differences were observed in expression levels of each factor between groups. However, 4 dogs in the chemotherapy-resistant group showed high expression of ABCB1. No significant difference was observed in the frequency of p53 mutation between groups. A possible association of ABCB1 with resistance to UW-25 was shown, but no uniform mechanism associated with drug resistance could be identified in dogs with lymphoma.

    PubMed

  • Yamazaki J, Takahashi M, Setoguchi A, Fujino Y, Ohno K, Tsujimoto H .  Monitoring of minimal residual disease (MRD) after multidrug chemotherapy and its correlation to outcome in dogs with lymphoma: a proof-of-concept pilot study. .  J Vet Intern Med24 ( 4 ) 897 - 903   2010年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Tumor cell burden in dogs with lymphoma cannot be assessed accurately by diagnostic evaluation during clinical complete remission (CR). Recent advances in polymerase chain reaction (PCR)-based methods enabled us to quantify minimal residual disease (MRD) in canine lymphoma. HYPOTHESIS/OBJECTIVES: To quantify MRD in dogs with lymphoma treated with multidrug chemotherapy and to correlate it with remission duration after chemotherapy. ANIMALS: Seventeen dogs with lymphoma that achieved CR by multidrug chemotherapy. METHODS: Rearranged immunoglobulin heavy chain or T-cell receptor gamma chain gene fragments from lymphoma cells were PCR amplified and sequenced to prepare clone-specific primers and probes for real-time PCR to quantify MRD. MRD in the peripheral blood was monitored during and at the end of a 25-week multidrug chemotherapy protocol. Correlation between MRD at the end of chemotherapy and remission duration after chemotherapy was analyzed. RESULTS: MRD gradually decreased after initiation of multidrug chemotherapy, reached a nadir as low as <0.019-1.0 cells/microL at weeks 4-17, and remained low or slightly increased until week 25. MRD at the end of chemotherapy was negatively correlated with remission duration from the end of chemotherapy to relapse. CONCLUSION AND CLINICAL IMPORTANCE: MRD could be an objective marker to indicate tumor cell burden in dogs with lymphoma even in clinical CR. MRD at the end of chemotherapy could be a prognostic factor to predict remission duration after chemotherapy.

    DOI: 10.1111/j.1939-1676.2010.0536.x

    PubMed

  • Tsuboi Masaya, Uchida Kazuyuki, Park Eun Sil, Kotera Yukiko, Seki Takahiro, Takahashi Masashi, Nakayama Hiroyuki .  Systemic T cell large granular lymphocyte lymphoma with multifocal white matter degeneration in the brain of a Japanese domestic cat. .  J Vet Med Sci72 ( 6 ) 795 - 799   2010年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A 10-year-old spayed female Japanese domestic cat exhibited clinical symptoms suggesting pancreatitis. One month later the cat exhibited Horner's syndrome and was euthanized. At necropsy, multiple neoplastic masses were found in the intestines, spleen, kidneys, urinary bladder, and lungs. On cytology, many neoplastic lymphocytic cells had fine to large cytoplasmic granules, suggesting large granular lymphocyte (LGL) lymphoma. Histopathological examinations revealed infiltrative proliferation of the neoplastic cells in almost organs. Immunohistochemically, the neoplastic cells were intensely positive for CD3 and granzyme B. In the brain, there were multifocal white matter lesions characterized by diffuse myelin loss with mild infiltration of the neoplastic cells. Based on these findings, the cat was diagnosed as LGL lymphoma presumptively of intestinal origin with systemic involvement.

    PubMed

  • Ishihara Masahiro, Fujino Yasuhito, Setoguchi Asuka, Takahashi Masashi, Nakashima Ko, Ohno Koichi, Tsujimoto Hajime .  Evaluation of prognostic factors and establishment of a prognostic scoring system for canine primary immune-mediated hemolytic anemia. .  J Vet Med Sci72 ( 4 ) 465 - 470   2010年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Clinical courses of primary immune-mediated hemolytic anemia (pIMHA) in dogs are highly variable, however, limited information is available to predict their accurate prognoses. To evaluate the prognostic significance of clinical factors and to propose a scoring system to predict prognoses, the medical records of seventy-one dogs with pIMHA were reviewed. Overall mortality rate of dogs with pIMHA was 39% and most of the dogs died within 3 months from diagnosis. Sex, body weight, seasonality, packed corpuscular volume (PCV), platelet count (PLT), total plasma protein (TP), blood urea nitrogen, albumin, total bilirubin, sodium ion, prothrombin time, and fibrin/fibrinogen degradation products before immunosuppressive treatment can influence on survival time in dogs with pIMHA. A prognostic scoring system using a combination of sex, seasonality, PCV, PLT and TP can be statistically significant for raising the accuracy of prognostic prediction. Using the scoring system for prognostication in dogs with pIMHA may enable veterinarians to predict a prognosis easily and accurately.

    PubMed

  • Fukushima Kenjiro, Ohno Koichi, Koshino-Goto Yuko, Uchida Kazuyuki, Nomura Kohji, Takahashi Masashi, Nakashima Ko, Fujino Yasuhito, Tsujimoto Hajime .  Sensitivity for the detection of a clonally rearranged antigen receptor gene in endoscopically obtained biopsy specimens from canine alimentary lymphoma. .  J Vet Med Sci71 ( 12 ) 1673 - 1676   2009年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Canine alimentary lymphoma is currently diagnosed on the basis of findings of cytological or histopathological examination. However, it is often difficult to histopathologically distinguish alimentary lymphoma from lymphocytic-plasmacytic enteritis. Recently, the application of polymerase chain reaction for antigen receptor gene rearrangement (PARR) has been reported. In the present study, we assess the sensitivity of PARR analysis in diagnosing canine alimentary lymphoma using endoscopically obtained biopsy specimens from 12 dogs that were histopathologically diagnosed as having lymphoma. The sensitivity of PARR analysis in diagnosing alimentary lymphoma was found to be 66.7%, which was lower than that of other lymphoid malignancies. A combination of histipathological examination and findings of PARR analysis may improve the diagnostic accuracy.

    PubMed

  • Kanemoto Hideyuki, Ohno Koichi, Sakai Manabu, Nakashima Ko, Takahashi Masashi, Fujino Yasuhito, Tsujimoto Hajime .  Blood hyaluronic acid as a marker for canine cirrhosis. .  J Vet Med Sci71 ( 9 ) 1251 - 1254   2009年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Blood hyaluronic acid (HA) concentration was measured in dogs with various liver diseases to determine its relationship with histological fibrosis of the liver. The blood HA concentration significantly increased in dogs with chronic liver diseases compared with extrahepatic diseases and control. Furthermore, the median blood HA concentration in dogs with liver cirrhosis (500 microg/l; range, 151-1970 microg/l) was significantly higher than dogs with non-cirrhotic liver diseases (153 microg/l; range, 15-477 microg/l). In histochemical analysis, HA was distributed primarily in the fibrotic area in dogs with chronic liver diseases. As a conclusion, the blood HA concentration was significantly increased in dogs with chronic liver diseases, especially those with cirrhosis. Measurement of the blood HA levels of dogs with suspected liver disease can be a useful diagnostic aid for canine cirrhosis.

    PubMed

  • Kanemoto Hideyuki, Ohno Koichi, Nakashima Ko, Takahashi Masashi, Fujino Yasuhito, Nishimura Ryohei, Tsujimoto Hajime .  Characterization of canine focal liver lesions with contrast-enhanced ultrasound using a novel contrast agent-sonazoid. .  Vet Radiol Ultrasound50 ( 2 ) 188 - 194   2009年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Contrast-enhanced ultrasound using Sonazoid, a novel contrast medium with a liver-specific Kupffer phase, was evaluated in canine focal liver lesions Twenty-five dogs with a liver mass were given intravenous Sonazoid, and the enhancement pattern in the arterial, portal, and parenchymal phase was characterized. An enhancement defect in the lesion in the parenchymal phase was observed in all malignant lesions, whereas only one of nine benign lesions had a filling defect. The diagnostic value of the presence of a filling defect for malignancy was statistically significant (100% sensitivity, 88.9% specificity, 94.1% positive predictive value, 100% negative predictive value), and was equal to that of hypoenhancement in the portal or delayed phase. The defect pattern (clear or irregular defect) was dependent (P < 0.05) on the types of malignancy (i.e., hepatocellular carcinoma and other types of malignancies). In the arterial phase, five of the six hepatocellular carcinomas had hypervascularity, whereas no other lesion was characterized by hypervascularity. In some dogs, additional lesions that could not be observed with conventional B-mode ultrasonography were detected in the parenchymal phase. The enhancement pattern of Sonazoid, especially in the parenchymal phase, has potential as a diagnostic tool for canine focal liver lesions.

    PubMed

  • Matsuki Naoaki, Takahashi Masashi, Yaegashi Masaya, Tamahara Satoshi, Ono Kenichiro .  Serial examinations of anti-GFAP autoantibodies in cerebrospinal fluids in canine necrotizing meningoencephalitis. .  J Vet Med Sci71 ( 1 ) 99 - 100   2009年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Canine necrotizing meningoencephalitis (NME) is characterized by autoantibodies against glial fibrillary acidic protein (GFAP) in cerebrospinal fluids (CSFs). To clarify the time-course changes in autoantibodies, serial examinations were conducted in three dogs with NME (two Pugs and a Pomeranian) that were treated by immunosuppressive therapy. The Pugs retained high autoantibody titers throughout the observation periods (146 and 813 days) and died with neurological signs. On the other hand, the Pomeranian switched from being positive for autoantibody to negative after day 580, and its NME seemed to be in clinical remission until death on day 1238. Therefore, the anti-GFAP autoantibodies can be detected over time in canine NME even during immunosuppressive therapies. However, the autoantibodies can also disappear within a certain period after onset.

    PubMed

  • Kanemoto Hideyuki, Ohno Koichi, Nakashima Ko, Takahashi Masashi, Fujino Yasuhito, Tsujimoto Hajime .  Vascular and Kupffer imaging of canine liver and spleen using the new contrast agent Sonazoid. .  J Vet Med Sci70 ( 11 ) 1265 - 1268   2008年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The new ultrasound contrast agent Sonazoid was injected in 5 healthy dogs, and the time course enhancement in liver parenchyma, portal vein, spleen, and renal cortex was evaluated. In liver parenchyma and spleen, sustained enhancement was observed from at least 8 to 15 min after injection with the peak at 45 sec (liver) and 20 sec (spleen), whereas in the portal vein and renal cortex, the time course enhancement dramatically decreased after the peak enhancement at 30 sec and 20 sec, respectively. No adverse effect was observed after Sonazoid injection in all the dogs that we examined. Contrast enhanced ultrasound using Sonazoid is believed to be useful for observing the parenchyma of canine liver and spleen.

    PubMed

  • Takeuchi Yoshinori, Matsuura Shinobu, Fujino Yasuhito, Nakajima Mayumi, Takahashi Masashi, Nakashima Ko, Sakai Yusuke, Uetsuka Koji, Ohno Koichi, Nakayama Hiroyuki, Tsujimoto Hajime .  Hypereosinophilic syndrome in two cats. .  J Vet Med Sci70 ( 10 ) 1085 - 1089   2008年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    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.

    PubMed

  • Nakamura Masaaki, Takahashi Masashi, Ohno Koichi, Koshino Akiko, Nakashima Ko, Setoguchi Asuka, Fujino Yasuhito, Tsujimoto Hajime .  C-reactive protein concentration in dogs with various diseases. .  J Vet Med Sci70 ( 2 ) 127 - 131   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    To investigate the clinical utility of C-reactive protein (CRP) determination in dogs, its plasma concentration was measured by a laser nephelometric method in 928 dogs with various diseases, and was compared with other inflammatory parameters. CRP concentration was elevated in various inflammatory diseases, this was most frequently observed in cases with neoplastic and immune-mediated diseases. All cases of pyometra, panniculitis, acute pancreatitis, polyarthritis, and hemangiosarcoma showed significantly elevated CRP concentrations. On the other hand, the CRP concentration was elevated only in few cases of neurological diseases such as epilepsy, meningoencephalitis, and hydrocephalus and endocrine diseases such as hypothyroidism, hyperadrenocorticism, and diabetes mellitus. Only a weak correlation was observed between the CRP concentration and white blood cell (WBC) counts (r=0.44) but no correlation with band neutrophil counts. There was no correlation between the CRP and albumin concentrations, but a weak negative correlation (r=-0.40) when excluding chronic intestinal diseases and nephrotic syndrome, which can cause protein loss. Thus, CRP can be useful to detect inflammations that cannot be detected by WBC and, or band neutrophil counts, suggesting that the examination of CRP concentration is essential as routine diagnostic test.

    PubMed

  • Suzuki Koretoki, Wakabayashi Haruna, Takahashi Masashi, Fukushima Kenjiro, Yabuki Akira, Endo Yasuyuki .  A Possible treatment strategy and clinical factors to estimate the treatment response in Bebesia gibsoni infection. .  J Vet Med Sci69 ( 5 ) 563 - 568   2007年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The effectiveness of combination therapy using clindamycin, metronidazole and doxycycline against canine babesiosis, and the usefulness of platelet count and the plasma C-reactive protein (CRP) concentration as an estimation factor for treatment, were evaluated in four dogs experimentally infected with Babesia gibsoni. The combination therapy successfully eliminated B. gibsoni in peripheral blood in 3 of 4 dogs, however the remaining dog showed obvious uncontrolled relapse after a temporary recovery. In addition, it was shown that CRP levels decreased in an inverse relationship to the recovery of packed cell volume and therefore CRP levels could be used as an optional clinical marker to estimate the response to treatment.

    PubMed

  • Takahashi Masashi, Une Ryoma, Fukushima Kenjiro, Fujiki Makoto, Misumi Kazuhiro, Miyoshi Noriaki, Endo Yasuyuki, Ohishi Akihiro, Akuzawa Masao .  Molecular cloning of canine nm23 cDNAs and their expression in normal and tumor tissues. .  J Vet Med Sci67 ( 8 ) 837 - 841   2005年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Two canine nm-23 cDNAs, designated as nm23-C1 and -C2, were isolated and characterized. Both have a putative open reading frame consisting of 459-bp encoding 152 amino acids and are highly similar to human, mouse and rat homologues. To understand the potential role of nm23-C1 and -C2 in the development of mammary gland tumors (MGT), we analyzed the mRNA expression in 14 MGT samples by RT-PCR. The samples were divided into categories according to their histopathology (benign/malignant) and metastasis. No significant difference in the mRNA expression levels of either nm23-C1 or -C2 were observed between the benign and malignant groups or the metastatic and non-metastatic groups. These results suggest that nm23-C1 and -C2 are not related to the establishment of malignancy and metastatic lesions in canine MGT cases.

    PubMed

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MISC

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)ファームプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)ファームプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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  • 腫瘍別プロトコル解説集 抗がん薬の組み合せを理解する!膀胱・前立腺癌

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  • 腫瘍別プロトコル解説集 抗がん薬の組み合せを理解する!組織球性肉腫

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  • 胸腺腫関連性剥離性皮膚炎が疑われた猫に対しシクロスポリンの単剤投与が有効であった1症例

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  • 肝臓腫瘍徹底攻略シンポジウム(3)肝臓腫瘍の診断と内科治療

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  • 消化器型リンパ腫 消化器型リンパ腫(高悪性度)

    高橋雅

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  • 基礎からおさえる呼吸器診療の最前線 第3回 鼻炎(犬・猫)および猫のウイルス性上部気道疾患

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  • 全血血栓形成観測システム(Total Thrombus Formation System:T-TAS)の血栓性疾患の評価

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  • 一般臨床医が知っておくべき化学療法の知識

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  • 一目でわかる症候シリーズ Vol.21 下痢 2)猫の胃腸のリンパ腫

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  • 新規血液凝固能検査としての全血血栓形成観測システム(Total Thrombus-Formation System:T-TAS)の臨床使用例

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  • 犬における血栓塞栓症の予防

    高橋雅

    MVM   27 ( 4 )   2018年

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  • 血液が固まり過ぎる(血栓症)

    高橋雅

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  • 脾臓摘出後に門脈血栓症を発症した犬の1例

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  • 【治療の前に!腫瘍随伴症候群を知る】おさえておくべき基礎知識

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    記述言語:日本語   出版者・発行元:(株)緑書房  

  • 【もっと知りたい!犬の膀胱移行上皮癌】ここに!注目 内科療法UP TO DATE

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    記述言語:日本語   出版者・発行元:(株)エデュワードプレス  

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    記述言語:日本語   出版者・発行元:(株)ファームプレス  

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    記述言語:日本語   出版者・発行元:(株)ファームプレス  

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  • 新規血液凝固能検査としての全血血栓形成観測システム(Total Thrombus Formation System:T-TAS)の臨床使用例

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  • 血栓塞栓症の診断と治療

    高橋雅

    日本獣医師会獣医学術学会年次大会講演要旨集   2016   2017年

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  • 犬の膀胱移行上皮癌 内科療法UP TO DATE

    高橋雅

    Veterinary Oncology   4 ( 3 )   2017年

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  • 治療の前に!腫瘍随伴症候群を知る おさえておくべき基礎知識

    高橋雅

    月刊CAP   ( 339 )   2017年

     詳細を見る

  • リンパ腫の治療ポイント,失敗する原因

    高橋雅

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  • フローサイトメトリー法を用いた抗赤血球抗体の検出

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講演・口頭発表等

  • 古澤 悠, 高橋 雅, 一二三達郎, 三好宣彰, 大和 修, 矢吹 映 .  犬の尿路移行上皮癌および前立腺癌における細胞診迅速蛍光抗体法を用いたCOX-2の検出 .  日本獣医臨床病理学会  2022年9月  日本獣医臨床病理学会

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    開催年月日: 2022年9月

    開催地:オンライン   国名:日本国  

  • 岩永 朋子, 市川 華帆, 齋藤 靖生, 高橋 雅, 福島 隆治, 三浦 直樹 .  新規血液凝固能検査としての全血血栓形成観測システム(Total Thrombus-Formation System:T-TAS)の臨床使用例 .  日本獣医師会雑誌  2018年11月  (公社)日本獣医師会

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    記述言語:日本語  

  • 岩永 朋子, 齋藤 靖生, 畑井 仁, 永里 朋香, 伊藤 隆史, 福島 隆治, 高橋 雅, 丸山 征郎, 三浦 直樹 .  LPS誘導ウサギDICモデルにおけるDIC初期の血小板の動向 血栓形成の可視化による検索 .  日本獣医学会学術集会講演要旨集  2020年10月  (公社)日本獣医学会

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    記述言語:日本語  

  • 岩永 朋子, 齋藤 靖生, 畑井 仁, 永里 朋香, 伊藤 隆史, 福島 隆治, 高橋 雅, 細川 和也, 三浦 直樹, 丸山 征郎 .  LPS投与ウサギモデルにおけるDIC前段階の血小板と白血球の可視化による解析 .  日本血栓止血学会誌  2021年5月  (一社)日本血栓止血学会

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    記述言語:日本語  

  • 遠藤 水月, 高橋 雅, 西田 英理子, 畑井 仁, 遠藤 泰之 .  赤血球内にカボット環を認めた急性白血病の猫の1例 .  日本獣医師会雑誌  2020年11月  (公社)日本獣医師会

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    記述言語:日本語  

  • 高橋 雅 .  血液が固まり過ぎる(血栓症) .  動物臨床医学会年次大会プロシーディング  2018年11月  動物臨床医学会

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    記述言語:日本語  

  • 高橋 雅 .  中枢神経症状を呈した急性骨髄性白血病の犬の1例 .  動物臨床医学会年次大会プロシーディング  2021年11月  動物臨床医学会

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    記述言語:日本語  

  • 高橋 雅 .  猫のオンコロジックエマージェンシー .  日本獣医がん学会招待

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    記述言語:日本語   会議種別:口頭発表(招待・特別)  

    開催地:オンライン   国名:日本国  

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共同研究・競争的資金等の研究

  • エクソソームとcfDNAを利用した腫瘍の超特異的&超高感度診断法の開発と臨床実証

    研究課題/領域番号:20K21375  2020年7月 - 2022年3月

    日本学術振興会  科学研究費助成事業 挑戦的研究(萌芽)  挑戦的研究(萌芽)

    三浦 直樹, 高橋 雅, 古澤 悠, 畑井 仁, 川原 幸一

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    配分額:6370000円 ( 直接経費:4900000円 、 間接経費:1470000円 )

  • 血球貪食性組織球肉腫は腫瘍なのか?マクロファージの自己認識の破綻からのアプローチ

    研究課題/領域番号:17K15381  2017年4月 - 2019年3月

    日本学術振興会  科学研究費助成事業 若手研究(B)  若手研究(B)

    高橋 雅

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    配分額:2860000円 ( 直接経費:2200000円 、 間接経費:660000円 )

    犬の血球貪食症候群と類似した特徴を有するヒトの血球貪食症候群ではCD47-SIRPαシグナルによる貪食抑制シグナルを破綻させることが発症と関与していることが報告されている。犬において血球貪食症候群の病態を明らかにするため、このCD47-SIRPαシグナルに関する基礎的検討を計画した。健康犬の各血球におけるCD47およびSIRPαの発現分布をフローサイトメトリーにより評価し、それぞれヒトの血球での発現と類似していることを明らかにした。さらに、抗体などの条件検討により免疫組織化学によって脾臓のマクロファージにおけるSIRPαの発現を評価することが可能となった。

  • イヌにおけるDIC診断の新規検査の検討

    研究課題/領域番号:16K08054  2016年10月 - 2020年3月

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    岩永 朋子, 福島 隆治, 齋藤 靖生, 丸山 征郎, 高橋 雅, 三浦 直樹

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    配分額:4160000円 ( 直接経費:3200000円 、 間接経費:960000円 )

    犬の新規血液凝固検査としてTotal Thrombus formation Analysis System(T-TAS)を用いて、初年度は健常コントロール群に対して、その再現性およびIn vivoおよびIn Vitroにて抗凝固薬の影響を判定した。次年度以降は周術期の出血リスクに関して、術前にAPTT,PTおよびT-TASを用いてそのリスクを評価し、周術期の実際の出血リスクを判定した。さらに、膵炎などの全身性炎症性疾患におけるDICリスクを血栓マーカーとの比較をおこなった。さらに循環器疾患における血小板凝集能に関して検討を行った。