Researcher Profile and Settings

Master

Affiliation (Master)

• Faculty of Veterinary Medicine　Veterinary Medicine　Clinical Sciences

Affiliation (Master)

• Faculty of Veterinary Medicine　Veterinary Medicine　Clinical Sciences

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Profile and Settings

Degree

• PhD(Veterinary Medicine)（Hokkaido University）
• Master of Veterinary Medical Science（Hokkaido University）

Profile and Settings

• Name (Japanese)

  Takiguchi

• Name (Kana)

  Mitsuyoshi

• Name

  200901022044782571

Achievement

Research Interests

• Chronic inflammation   自己免疫疾患   超音波診断   画像診断   Veterinary Diagnostic imaging   Autoimmune Diseases   Diagnostic Ultrasound   Diagnostic imaging   

Research Areas

• Life sciences / Radiology
• Life sciences / Allergies and connective tissue disease
• Life sciences / Veterinary medicine

Research Experience

• 2021/04 Hokkaido University Faculty of Veterinary Medicine
• 2017/04 北海道大学大学院獣医学研究院 教授
• 2007/10 Hokkaido University Graduate School of Veterinary Medicine Professor
• 2004/10 - 2007/09 Rakuno Gakuen University School of Veterinary Medicine Professor
• 2004/06 Hokkaido University Graduate School of Veterinary Medicine
• 2000 - 2004 Lecturer, Graduate School of Veterinary Medicine,
• 2004 - Professor, School of Veterinary Medicine, Rakuno gakuen Univ.
• 2004 - Assistant Professor, Graduate School of Veterinary Medicine,
• 2000/07 Hokkaido University Graduate School of Veterinary Medicine
• 1995 - 2000 Instractor, Graduate School of Veterinary Medicine
• 1996/03 - 1996/08 文部省在外研究員
• 1995/04 Hokkaido University Graduate School of Veterinary Medicine
• 1993 - 1995 Instractor, Faculty of Veterinary Medicine, Hokkaido Univ.
• 1993/10 Hokkaido University School of Veterinary Medicine
• 1992/06 The Ohio State University
• 1989/04 - 1992/05 Yokohama Animal Medical Center
• Hokkaiddo Univ.

Education

• 1987/04 - 1989/03  Hokkaido University  Graduate School of Veterinary Medicine
• 1983/04 - 1987/03  Hokkaido University  School of Veterinary Medicine  Veterinary Medicine

Awards

• 2004 日本小動物獣医学会北海道獣医師会長賞
  
• 1999 平成11年度日本小動物獣医学会(北海道)獣医師会長賞
  
• 1999 日本獣医学会大会長賞 獣医学奨励賞
  
• 1998 日本獣医学会大会長賞 獣医学奨励賞
  
• 1997 平成9年度日本小動物獣医学会(北海道)地区学会長賞
  

Published Papers

• Reference intervals for urinary metanephrines to creatinine ratios in dogs in Japan.

  Noriyuki Nagata, Yoshinori Ikenaka, Hiryu Sawamura, Noboru Sasaki, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 2024/09/03 

  The urinary normetanephrine to creatinine ratio (uNMN/Cr) and urinary metanephrine to creatinine ratio (uMN/Cr) are commonly evaluated for the diagnosis of pheochromocytomas (PCC) in dogs. This study aimed to establish reference intervals for uNMN/Cr and uMN/Cr in 56 healthy dogs in Japan and to investigate the effect of urine collection methods on these measurements in 15 non-PCC dogs. The upper limits of reference intervals for uNMN/Cr and uMN/Cr were 124.4 nmol/mmol (90% confidence interval [CI] 107.7-137.0) and 121.1 nmol/mmol (90% CI 102.8-145.1), respectively. Both uNMN/Cr and uMN/Cr were significantly higher when urine was collected in the hospital compared to at home. Several factors, including the method of urine collection, should be considered when utilizing these reference intervals.
• Case report: Echinococcus multilocularis infection in a dog showing gastrointestinal signs in Hokkaido, Japan

  Izumi Kida, Naoki Hayashi, Nozomu Yokoyama, Noriyuki Nagata, Kazuyoshi Sasaoka, Noboru Sasaki, Keitaro Morishita, Kensuke Nakamura, Hirokazu Kouguchi, Kinpei Yagi, Ryo Nakao, Mitsuyoshi Takiguchi, Nariaki Nonaka

  Frontiers in Veterinary Science 11 2024/06/13 [Refereed][Not invited]
   

  Echinococcus multilocularis is a cestode that causes human alveolar echinococcosis, a lethal zoonotic disease distributed in the northern hemisphere. The life cycle of this parasite is maintained in nature by voles as intermediate hosts and foxes as definitive hosts in Hokkaido, Japan. Although dogs are also susceptible to the parasite, the infection has been considered typically asymptomatic. We report the detection of E. multilocularis eggs in the diarrheal feces of a dog with chronic gastrointestinal signs, which disappeared after anthelmintic treatment. The mitochondrial genome sequence constructed by sequencing of the overlapping PCRs using DNA from the eggs was identical to the most predominant haplotype previously reported in red foxes in Hokkaido. This case highlights that Echinococcus infection should be considered as a differential diagnosis for diarrheal dogs in the disease endemic areas. Further efforts are needed to accumulate parasite genotypes in domestic dogs as well as humans to assess the risk of human infection from dogs.
• Urinary steroid profiling using liquid chromatography-tandem mass spectrometry for the diagnosis of canine Cushing's syndrome.

  N Nagata, H Sawamura, Y Ikenaka, K Morishita, K Hosoya, N Sasaki, K Nakamura, M Takiguchi

  Veterinary journal (London, England : 1997) 106151 - 106151 2024/05/29 [Refereed][Not invited]
   

  Serum cortisol measurements by chemiluminescence enzyme immunoassay (CLEIA) are widely used to diagnose hypercortisolism (HC) or Cushing's syndrome in dogs. However, they are associated with problems such as the need for multiple blood collections under stressful conditions or cross-reactivity between hormones. Therefore, a less invasive and more accurate diagnostic method is required. This study aimed to develop a urinary steroid profile analysis method using liquid chromatography-tandem mass spectrometry (LC/MS/MS) and to evaluate its clinical usefulness. Sixty-five healthy dogs and 38 dogs with suspected HC were included in the study. Using LC/MS/MS, the levels of 11 steroid hormones in the urine were determined. We established the upper limit of the reference interval for each urinary steroid-to-creatinine ratio and evaluated their diagnostic performances. The levels of the five steroid hormones were significantly higher in the 14 dogs with HC than in the 24 dogs with mimicking HC and 65 healthy dogs. The urinary corticosterone-to-creatinine ratio showed the highest diagnostic accuracy (area under the curve, 0.96). A significant correlation was seen between urinary cortisol concentrations measured by LC/MS/MS and CLEIA (rs = 0.88, P <0.001), although the CLEIA measurements were significantly higher than the LC/MS/MS measurements (P <0.001). LC/MS/MS-based urinary steroid profiles are a promising tool for diagnosing canine HC.
• Antimicrobial resistance and self-reported hand hygiene awareness before and after an infection prevention and control programme: a 7-year analysis in a small animal veterinary teaching hospital

  K. Sasaoka, T. Sato, K. Morishita, K. Hosoya, N. Yokoyama, T. Sato, M. Horiuchi, M. Takiguchi

  The Veterinary Journal 106154 - 106154 1090-0233 2024/05 [Refereed][Not invited]
  
• A de novo nonsense variant in the DMD gene associated with X-linked dystrophin-deficient muscular dystrophy in a cat.

  Nozomu Yokoyama, Yuki Matsumoto, Takahisa Yamaguchi, Kazuki Okada, Ryohei Kinoshita, Genya Shimbo, Hisashi Ukawa, Ryuga Ishii, Kensuke Nakamura, Jumpei Yamazaki, Mitsuyoshi Takiguchi

  Journal of veterinary internal medicine 2024/04/13 [Refereed]
   

  BACKGROUND: X-linked dystrophin-deficient muscular dystrophy (MD) is a form of MD caused by variants in the DMD gene. It is a fatal disease characterized by progressive weakness and degeneration of skeletal muscles. HYPOTHESIS/OBJECTIVES: Identify deleterious genetic variants in DMD by whole-genome sequencing (WGS) using a next-generation sequencer. ANIMALS: One MD-affected cat, its parents, and 354 cats from a breeding colony. METHODS: We compared the WGS data of the affected cat with data available in the National Center for Biotechnology Information database and searched for candidate high-impact variants by in silico analyses. Next, we confirmed the candidate variants by Sanger sequencing using samples from the parents and cats from the breeding colony. We used 2 genome assemblies, the standard felCat9 (from an Abyssinian cat) and the novel AnAms1.0 (from an American Shorthair cat), to evaluate genome assembly differences. RESULTS: We found 2 novel high-impact variants: a 1-bp deletion in felCat9 and an identical nonsense variant in felCat9 and AnAms1.0. Whole genome and Sanger sequencing validation showed that the deletion in felCat9 was a false positive because of misassembly. Among the 357 cats, the nonsense variant was only found in the affected cat, which indicated it was a de novo variant. CONCLUSION AND CLINICAL IMPORTANCE: We identified a de novo variant in the affected cat and next-generation sequencing-based genotyping of the whole DMD gene was determined to be necessary for affected cats because the parents of the affected cat did not have the risk variant.
• Interventricular inflow time difference assessed by dual pulsed-wave Doppler echocardiography in dogs with myxomatous mitral valve disease

  T. Morita, K. Nakamura, T. Osuga, S. Kawamoto, M. Takiguchi

  Journal of Veterinary Cardiology 51 24 - 34 1760-2734 2024/02 [Refereed][Not invited]
  
• Use of genome-wide DNA methylation analysis to identify prognostic CpG site markers associated with longer survival time in dogs with multicentric high-grade B-cell lymphoma.

  Yong Bin Teoh, Teita Ishizaki, Yumiko Kagawa, Shoko Yokoyama, Jaroslav Jelinek, Yuki Matsumoto, Hirotaka Tomiyasu, Hajime Tsujimoto, Mitsuyoshi Takiguchi, Jumpei Yamazaki

  Journal of veterinary internal medicine 38 (1) 316 - 325 2024 [Refereed]
   

  BACKGROUND: DNA methylation analysis might identify prognostic CpG sites in CHOP-treated dogs with multicentric high-grade B-cell lymphoma (MHGL) with heterogenous prognosis. OBJECTIVE: To identify prognostic CpG sites of MHGL through genome-wide DNA methylation analysis with pyrosequencing validation. ANIMALS: Test group: 24 dogs. Validation group: 100 dogs. All client-owned dogs were diagnosed with MHGL and treated with CHOP chemotherapy. METHODS: Cohort study. DNA was extracted from lymph node samples obtained via FNA. Genome-wide DNA methylation analysis using Digital Restriction Enzyme Analysis of Methylation (DREAM) was performed on the test group to identify differentially methylated CpG sites (DMCs). Bisulfite pyrosequencing was used to measure methylation status of candidate DMCs in the validation group. Median survival times (MST) were analyzed using Kaplan-Meier (log-rank) product limit method. RESULTS: DREAM analyzed 101 576 CpG sites. Hierarchical clustering of 16 262 CpG sites in test group identified group with better prognosis (MST = 55-477 days vs 10-301 days, P = .007). Volcano plot identified 1371 differentially methylated CpG sites (DMCs). DMC near the genes of FAM213A (DMC-F) and PHLPP1 (DMC-P) were selected as candidates. Bisulfite-pyrosequencing performed on validation group showed group with methylation level of DMC-F < 40% had favorable prognosis (MST = 11-1072 days vs 8-1792 days, P = .01), whereas group with the methylation level combination of DMC-F < 40% plus DMC-P < 10% had excellent prognosis (MST = 18-1072 days vs 8-1792 days, P = .009). CONCLUSION AND CLINICAL IMPORTANCE: Methylation status of prognostic CpG sites delineate canine MGHL cases with longer MST, providing owners with information on expectations of potential improved treatment outcomes.
• Adrenocortical hypoperfusion detected by contrast-enhanced ultrasound in a dog with trilostane-induced hypoadrenocorticism.

  N Nagata, K Aoshima, K Nakamura, M Takiguchi

  The Journal of small animal practice 64 (11) 722 - 726 2023/11 [Refereed]
   

  A 12-year-old neutered male Chihuahua dog was diagnosed with pituitary-dependent hypercortisolism and treated with trilostane. Eighty-nine days later, the dog showed lethargy accompanied by hyponatraemia and hyperkalaemia. Hypoadrenocorticism due to trilostane was suspected, but the result of the adrenocorticotropic hormone stimulation test was not conclusive. Contrast-enhanced ultrasound showed loss of adrenocortical blood flow in both adrenal glands, indicating adrenocortical hypoperfusion and isolated hypoadrenocorticism. Treatment with fludrocortisone acetate improved the condition and electrolyte abnormalities. Thirteen months later, the dog showed alopecia, and an adrenocorticotropic hormone stimulation test revealed increased cortisol concentration, indicating hypercortisolism recurrence. The dog died due to progressive deterioration 22 months after the initial presentation. Post-mortem examination revealed focally extensive necrosis with marked calcification in the parenchyma of the adrenal glands and regeneration of the cells in the zona fasciculata with severe fibrosis. Adrenocortical hypoperfusion detected by contrast-enhanced ultrasound can support the diagnosis of adrenal necrosis and hypoadrenocorticism.
• The role of heat shock protein 90 in the proliferation of Babesia gibsoni in vitro.

  Hiroka Tsutsumi, Moeko Abe, Naohiro Uchida, Mitsuyoshi Takiguchi, Masahiro Yamasaki

  Experimental parasitology 251 108567 - 108567 2023/08 [Refereed]
   

  The present study investigated the role of heat shock protein 90 (HSP90) in the proliferation and survival of Babesia gibsoni in vitro. To detect the effect on the entry of B. gibsoni into host erythrocytes, the parasite was incubated with an antibody against B. gibsoni HSP90 (BgHSP90) for 24 h. The results of this experiment demonstrated that both the incorporation of [3H]hypoxanthine into the nucleic acids of B. gibsoni and the number of parasites were not altered, indicating that an anti-BgHSP90 antibody did not directly inhibit the entry of the parasite into erythrocytes. Moreover, two HSP90 inhibitors, geldanamycin (GA) and tanespimycin (17-AAG), were used to evaluate the function of BgHSP90. GA and 17-AAG decreased both the incorporation of [3H]hypoxanthine and the number of infected erythrocytes, suggesting that BgHSP90 plays important roles in DNA synthesis and the proliferation of B. gibsoni. The effect of 17-AAG on the parasites was weaker than that of GA. Additionally, the effect of GA on the survival and superoxide generation of canine neutrophils was assessed. The survival of canine neutrophils was not affected. The superoxide generation was strongly suppressed by GA. This result indicated that GA inhibited the function of canine neutrophils. Additional studies are necessary to elucidate the role of BgHSP90 in the proliferation of the parasite.
• Diverse genome-wide DNA methylation alterations in canine hepatocellular tumours.

  Yu Asari, Jumpei Yamazaki, Oo Thandar, Tamami Suzuki, Keisuke Aoshima, Kyosuke Takeuchi, Ryohei Kinoshita, Sangho Kim, Kenji Hosoya, Teita Ishizaki, Yumiko Kagawa, Jaroslav Jelinek, Shoko Yokoyama, Noboru Sasaki, Hiroshi Ohta, Kensuke Nakamura, Mitsuyoshi Takiguchi

  Veterinary medicine and science 2023/07/22 [Refereed]
   

  BACKGROUND: Canine hepatocellular tumours (HCTs) are common primary liver tumours. However, the exact mechanisms of tumourigenesis remain unclear. Although some genetic mutations have been reported, DNA methylation alterations in canine HCT have not been well studied. OBJECTIVES: In this study, we aimed to analyse the DNA methylation status of canine HCT. METHODS: Tissues from 33 hepatocellular carcinomas, 3 hepatocellular adenomas, 1 nodular hyperplasia, 21 non-tumour livers from the patients and normal livers from 5 healthy dogs were used. We analysed the DNA methylation levels of 72,367 cytosine-guanine dinucleotides (CpG sites) in all 63 samples. RESULTS AND CONCLUSIONS: Although a large fraction of CpG sites that were highly methylated in the normal liver became hypomethylated in tumours from most patients, we also found some patients with less remarkable change or no change in DNA methylation. Hierarchical clustering analysis revealed that 32 of 37 tumour samples differed from normal livers, although the remaining 5 tumour livers fell into the same cluster as normal livers. In addition, the number of hypermethylated genes in tumour livers varied among tumour cases, suggesting various DNA methylation patterns in different tumour groups. However, patient and clinical parameters, such as age, were not associated with DNA methylation status. In conclusion, we found that HCTs undergo aberrant and diverse patterns of genome-wide DNA methylation compared with normal liver tissue, suggesting a complex epigenetic mechanism in canine HCT.
• An inflammatory bowel disease-associated SNP increases local thyroglobulin expression to develop inflammation in miniature dachshunds

  Yong Bin Teoh, Jing-Jing Jiang, Takeshi Yamasaki, Noriyuki Nagata, Toshiki Sugawara, Rie Hasebe, Hiroshi Ohta, Noboru Sasaki, Nozomu Yokoyama, Kensuke Nakamura, Yumiko Kagawa, Mitsuyoshi Takiguchi, Masaaki Murakami

  Frontiers in Veterinary Science 10 2023/07/14 [Refereed]
   

  Inflammatory colorectal polyp (ICRP) in miniature dachshunds (MDs) is a chronic inflammatory bowel disease (IBD) characterized by granulomatous inflammation that consists of neutrophil infiltration and goblet cell hyperplasia in the colon. Recently, we identified five MD-associated single-nucleotide polymorphisms (SNPs), namely PLG, TCOF1, TG, COL9A2, and COL4A4, by whole-exome sequencing. Here, we investigated whether TG c.4567C&gt;T (p.R1523W) is associated with the ICRP pathology. We found that the frequency of the T/T SNP risk allele was significantly increased in MDs with ICRP. In vitro experiments showed that TG expression in non-immune cells was increased by inducing the IL-6 amplifier with IL-6 and TNF-α. On the other hand, a deficiency of TG suppressed the IL-6 amplifier. Moreover, recombinant TG treatment enhanced the activation of the IL-6 amplifier, suggesting that TG is both a positive regulator and a target of the IL-6 amplifier. We also found that TG expression together with two NF-κB targets, IL6 and CCL2, was increased in colon samples isolated from MDs with the T/T risk allele compared to those with the C/C non-risk allele, but serum TG was not increased. Cumulatively, these results suggest that the T/T SNP is an expression quantitative trait locus (eQTL) of TG mRNA in the colon, and local TG expression triggered by this SNP increases the risk of ICRP in MDs via the IL-6 amplifier. Therefore, TG c.4567C&gt;T is a diagnostic target for ICRP in MDs, and TG-mediated IL-6 amplifier activation in the colon is a possible therapeutic target for ICRP.
• Successful treatment of sclerosing encapsulating peritonitis in a cat using bioresorbable hyaluronate-carboxymethylcellulose membrane after surgical adhesiolysis and long-term prednisolone

  Nozomu Yokoyama, Ryohei Kinoshita, Hiroshi Ohta, Kazuki Okada, Genya Shimbo, Kazuyoshi Sasaoka, Noriyuki Nagata, Noboru Sasaki, Keitaro Morishita, Kensuke Nakamura, Yumiko Kagawa, Mitsuyoshi Takiguchi

  Journal of Feline Medicine and Surgery Open Reports 9 (2) 2055-1169 2023/07 [Refereed]
   

  Case summary An 8-year-old neutered male domestic shorthair indoor cat was presented with an 8-week history of intermittent vomiting, anorexia and weight loss that had been unresponsive to supportive treatment. Abdominal ultrasound revealed plication of the small intestine and fluid accumulation proximal to the lesion, and a linear foreign body was suspected. An exploratory celiotomy showed cocoon-like encapsulation of the entire intestine. Surgical adhesiolysis and full-thickness biopsy were performed, and histopathologic examination revealed mild thickening of the visceral peritoneum with fibrin deposition, as well as mild neutrophil and lymphocyte infiltration. These findings were compatible with sclerosing encapsulating peritonitis (SEP). The cat recovered well postoperatively and was discharged the next day. Prednisolone was administered for 7 weeks to prevent recurrence of SEP. Five months after surgery, the cat was re-presented with anorexia and chronic vomiting. Based on the clinical examination findings, recurrent SEP was suspected. At the second surgery, surgical adhesiolysis was repeated and a bioresorbable hyaluronate-carboxymethylcellulose membrane was used to cover the serosal surface and thus prevent adhesion formation. Histopathologic findings of the peritoneal biopsy specimen confirmed SEP. Long-term prednisolone treatment (1 mg/kg for the first dose and 0.5 mg/kg every 48 h for maintenance) was administered postoperatively. The cat survived for more than 1239 days without recurrence. Relevance and novel information To our knowledge, this is the first report of SEP in a cat with long-term survival. The use of a bioresorbable hyaluronate-carboxymethylcellulose membrane and long-term prednisolone treatment may have prevented short-term and long-term recurrence, respectively, in this case.
• Investigation of the therapeutic effects, predictors, and complications of long-term immunosuppressive therapy in dogs with precursor-targeted immune-mediated anemia.

  Mei Sugawara-Suda, Keitaro Morishita, Yuto Iwanaga, Jumpei Yamazaki, Yumiko Kagawa, Nozomu Yokoyama, Noboru Sasaki, Hiroshi Ohta, Kensuke Nakamura, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 85 (7) 695 - 701 2023/07/01 [Refereed]
   

  Dogs with precursor-targeted immune-mediated anemia (PIMA) are commonly treated with immunosuppressive therapy, but information on predictors of treatment response and response time is limited. Therefore, we retrospectively investigated predictive factors that influenced the treatment response and duration required to observe a response in dogs with PIMA receiving continuous immunosuppressive therapies for more than 105 days. Of 50 client-owned dogs that developed PIMA, 27 were included in this study, of which 18 were responders and 9 were non-responders to immunosuppressive therapies. Sixteen of the 18 responders responded to treatment within 60 days and the remaining 2 responded at 93 and 126 days, respectively. We found that an erythroid-maturation ratio of <0.17 may be a useful predictor for treatment response. In addition, complications of immunosuppressive therapies were investigated further in 50 dogs. Pancreatitis (n=4) and pneumonia (3) occurred over the entire treatment period, and infections such as abscesses (3) tended to be more common in dogs on an extended period of immunosuppressive therapy. These findings may be helpful when planning for the initial treatment and may provide evidence for informed consent about potential comorbidities throughout the treatment course.
• Effectiveness of 2‐dimensional shear wave elastography for noninvasive and reliable estimation of right atrial pressure in dogs with induced volume overload

  Masahiro Tamura, Hiroshi Ohta, Tatsuyuki Osuga, Mitsuyoshi Takiguchi

  Journal of Veterinary Internal Medicine 0891-6640 2023/04/10 [Refereed]
  
• Evaluation of responses to immunosuppressive therapy in dogs with suspected non‐regenerative immune‐mediated anaemia: 11 cases (2012‐2018)

  K. Morishita, M. Sugawara‐Suda, J. Yamazaki, N. Sasaki, K. Nakamura, H. Ohta, M. Takiguchi

  Journal of Small Animal Practice 64 527  0022-4510 2023/04/07 [Refereed]
  
• Zoobiquity experiments show the importance of the local MMP9-plasminogen axis in inflammatory bowel diseases in both dogs and patients.

  Takeshi Yamasaki, Noriyuki Nagata, Toru Atsumi, Rie Hasebe, Yuki Tanaka, Izuru Ohki, Shimpei Kubota, Yuta Shinohara, Yong Bin Teoh, Nozomu Yokoyama, Noboru Sasaki, Kensuke Nakamura, Hiroshi Ohta, Takehiko Katsurada, Yoshihiro Matsuno, Shintaro Hojyo, Shigeru Hashimoto, Mitsuyoshi Takiguchi, Masaaki Murakami

  International immunology 35 (7) 313 - 326 2023/03/18 [Refereed]
   

  Using a zoobiquity concept, we directly connect animal phenotypes to a human disease mechanism: the reduction of local plasminogen levels caused by matrix metalloproteinase-9 (MMP9) activity is associated with the development of inflammation in the intestines of dogs and patients with inflammatory bowel disease. We first investigated inflammatory colorectal polyps (ICRPs), which are a canine gastrointestinal disease characterized by the presence of idiopathic chronic inflammation, in Miniature Dachshund (MD), and found 31 missense disease-associated SNPs by whole-exome sequencing. We sequenced them in 10 other dog breeds and found five, PLG, TCOF1, TG, COL9A2, and COL4A4, only in MD. We then investigated two rare and breed-specific missense SNPs (T/T SNPs), PLG: c.477G>T and c.478A>T, and found that ICRPs with the T/T SNP risk-alleles showed less intact plasminogen and plasmin activity in the lesions compared to ICRPs without the risk-alleles but no differences in serum. Moreover, we show that MMP9, which is a NF-κB target, caused the plasminogen reduction and that intestinal epithelial cells expressing plasminogen molecules were colocalized with epithelial cells expressing MMP9 in normal colons with the risk-alleles. Importantly, MMP9 expression in patients with ulcerous colitis or Crohn's disease also colocalized with epithelial cells showing enhanced NF-κB activation and less plasminogen expression. Overall, our zoobiquity experiments showed that MMP9 induces the plasminogen reduction in intestine, contributing to the development of local inflammation and suggesting the local MMP9-plasminogen axis is a therapeutic target in both dogs and patients. Therefore, zoobiquity-type experiments could bring new perspectives for biomarkers and therapeutic targets.
• Prognostic value of left atrial stiffness estimated using echocardiography in dogs with myxomatous mitral valve disease

  T. Osuga, K. Kuroda, T. Morita, N. Sasaki, K. Nakamura, M. Takiguchi

  Journal of Veterinary Cardiology 45 15 - 26 1760-2734 2023/02 [Refereed]
   

  Introduction: No studies have investigated the prognostic value of left atrial stiffness (LASt) estimated using echocardiography in dogs with myxomatous mitral valve disease (MMVD). Animals: 72 dogs had MMVD and 46 dogs were healthy dogs. Materials and Methods: Clinical retrospective cohort study. The survival information of MMVD dogs that underwent echocardiographic examination was obtained. The peak velocities of early diastolic transmitral flow (E) and mitral annular motion as determined by pulsed wave Doppler (E′) were determined. The left atrial reservoir strain (εS) was determined by two-dimensional speckle tracking echocardiography of the left atrium. The LASt was estimated by the formula: E/E′/εS. The 95% prediction interval of LASt was computed from the data of the healthy dogs. Results: 17 dogs having MMVD died of cardiac-related causes with 55 MMVD dogs censored. The MMVD dogs with LASt > its 95% upper prediction limit (LASt > 0.56; n = 26; median survival time, 484 days; 95% confidence interval, 283 days-indeterminable) had shorter survival times (P<0.001) than those with LASt ≤ its 95% upper prediction limit (LASt ≤ 0.56; n = 46; median survival time, >1112 days; 95% confidence interval, indeterminable). Multivariable Cox's proportional hazard analysis demonstrated that the ratio of the left atrial dimension to the aortic annulus dimension and LASt were independent predictors of cardiac-related death among conventional echocardiographic indices, εS and LASt in MMVD dogs. Conclusions: In dogs with MMVD, increased LASt estimated using echocardiography is an independent predictor of cardiac-related death, and LASt can be more useful for prognostication than εS.
• Transcriptome and proteome analysis of dogs with precursor targeted immune-mediated anemia treated with splenectomy.

  Mei Sugawara-Suda, Keitaro Morishita, Osamu Ichii, Takashi Namba, Keisuke Aoshima, Yumiko Kagawa, Sangho Kim, Kenji Hosoya, Nozomu Yokoyama, Noboru Sasaki, Kensuke Nakamura, Jumpei Yamazaki, Mitsuyoshi Takiguchi

  PloS one 18 (5) e0285415  2023 [Refereed]
   

  Precursor-targeted immune-mediated anemia (PIMA) in dogs is characterized by persistent non-regenerative anemia and ineffective erythropoiesis, and it is suspected to be an immune-mediated disease. Most affected dogs respond to immunosuppressive therapies; however, some are resistant. In this study, we carried out splenectomy as an alternative therapy for refractory PIMA in dogs, and analyzed gene expression levels in the spleen of dogs with or without PIMA and in serum before and after splenectomy. A total of 1,385 genes were found to express differentially in the spleens from dogs with PIMA compared with healthy dogs by transcriptome analysis, of which 707 genes were up-regulated, including S100A12, S100A8, and S100A9 that are linked directly to the innate immune system and have been characterized as endogenous damage-associated molecular patterns. Furthermore, immunohistochemistry confirmed that S100A8/A9 protein expression levels were significantly higher in dogs with PIMA compared with those in healthy dogs. A total of 22 proteins were found to express differentially between the serum samples collected before and after splenectomy by proteome analysis, of which 12 proteins were up-regulated in the samples before. The lectin pathway of complement activation was identified by pathway analysis in pre-splenectomy samples. We speculated that S100A8/9 expression may be increased in the spleen of dogs with PIMA, resulting in activation of the lectin pathway before splenectomy. These findings further our understanding of the pathology and mechanisms of splenectomy for PIMA.
• Biological effects related to exposure to polychlorinated biphenyl (PCB) and decabromodiphenyl ether (BDE-209) on cats

  Khidkhan K, Mizukawa H, Ikenaka Y, Nakayama SMM, Nomiyama K, Yokoyama N, Ichii O, Takiguchi M, Tanabe S, Ishizuka M

  PLOS ONE 18 (1) e0277689  2023/01 [Refereed]
   

  As an animal familiar to humans, cats are considered to be sensitive to chemicals; cats may be exposed to polychlorinated biphenyls (PCBs) and decabromodiphenyl ether (BDE-209) from indoor dust, household products, and common pet food, leading to adverse endocrine effects, such as thyroid hormone dysfunction. To elucidate the general biological effects resulting from exposure of cats to PCBs and PBDEs, cats were treated with a single i.p. dose of a principal mixture of 12 PCBs and observed for a short-term period. Results revealed that the testis weight, serum albumin, and total protein of the treated group decrease statistically in comparison with those in the control group. The negative correlations suggested that the decrease in the total protein and albumin levels may be disturbed by 4'OH-CB18, 3'OH-CB28 and 3OH-CB101. Meanwhile, the serum albumin level and relative brain weight decreased significantly for cats subjected to 1-year continuous oral administration of BDE-209 in comparison to those of control cats. In addition, the subcutaneous fat as well as serum high-density lipoprotein (HDL) and triglycerides (TG) levels increased in cats treated with BDE-209 and down-regulation of stearoyl-CoA desaturase mRNA expression in the liver occurred. These results suggested that chronic BDE-209 treatment may restrain lipolysis in the liver, which is associated with lipogenesis in the subcutaneous fat. Evidence of liver and kidney cell damage was not observed as there was no significant difference in the liver enzymes, blood urea nitrogen and creatinine levels between the two groups of both experiments. To the best of our knowledge, this is the first study that provides information on the biochemical effects of organohalogen compounds in cats. Further investigations on risk assessment and other potential health effects of PCBs and PBDEs on the reproductive system, brain, and lipid metabolism in cats are required.
• An experimental challenge model for Leishmania donovani in beagle dogs, showing a similar pattern of parasite burden in the peripheral blood and liver.

  Hiroya Konno, Nozomu Yokoyama, Yu Tamura, Keisuke Aoshima, Ryo Nakao, Mitsuyoshi Takiguchi, Ken Katakura

  Parasitology research 121 (12) 3569 - 3579 2022/12 [Refereed]
   

  Leishmania donovani and Leishmania infantum are closely related species. However, the former is considered the causative agent for anthroponotic visceral leishmaniasis (AVL), while the latter is known to be responsible for zoonotic visceral leishmaniasis (ZVL) with dogs as the main reservoir host. Although molecular detection of L. donovani from naturally infected dogs has been reported in AVL endemic areas, the experimental infection of dogs with this species is very limited. Here, we constructed an experimental canine visceral leishmaniasis (CVL) model with L. donovani infection using beagle dogs. During an observation period of 8 months after parasite inoculation, few clinical symptoms were observed in the three inoculated dogs. The overall hematological and biochemical data of the dogs showed normal levels, and there were no remarkable changes in the peripheral CD4+, CD8+, CD25+, or FoxP3+ T cell populations. Liver biopsy sampling was conducted to monitor the parasite burden in the liver. A similar pattern of the amount of mitochondrial kinetoplast DNA was observed in the peripheral blood and liver by real-time PCR analysis. In addition, parasite antigens were detected from the liver biopsy sections by immunohistochemical analysis, further supporting the existence of parasites in the liver. These results showed a subclinical CVL model for L. donovani in beagle dogs with a similar kinetics of parasite burden in the peripheral blood and liver.
• Health impact assessment of pet cats caused by organohalogen contaminants by serum metabolomics and thyroid hormone analysis.

  Kei Nomiyama, Yasuo Yamamoto, Akifumi Eguchi, Hiroyuki Nishikawa, Hazuki Mizukawa, Nozomu Yokoyama, Osamu Ichii, Mitsuyoshi Takiguchi, Shouta M M Nakayama, Yoshinori Ikenaka, Mayumi Ishizuka

  The Science of the total environment 842 156490 - 156490 2022/10/10 [Refereed]
   

  Companion animals are in close contact with the human surroundings, and there is growing concern about the effects of harmful substances on the health of pet cats. In this study, we investigated the potential health effects of organohalogen compounds (OHCs) on thyroid hormone (TH) homeostasis and metabolomics in Japanese pet cats. There was a significant negative correlation between concentrations of several contaminants, such as polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), hydroxylated PCBs (OH-PCBs), hydroxylated PBDEs (OH-PBDEs), and THs in cat serum samples. These results suggested that exposure to OHCs causes a decrease in serum TH levels in pet cats. In this metabolomics study, each exposure level of parent compounds (PCBs and PBDEs) and their hydroxylated compounds (OH-PCBs and OH-PBDEs) were associated with their own unique primary metabolic pathways, suggesting that parent and phenolic compounds exhibit different mechanisms of action and biological effects. PCBs were associated with many metabolic pathways, including glutathione and purine metabolism, and the effects were replicated in in-vivo cat PCB administration studies. These results demonstrated that OHC exposure causes chronic oxidative stress in pet cats. PBDEs were positively associated with alanine, aspartate, and glutamate metabolism. Due to the chronic exposure of cats to mixtures of these contaminants, the combination of their respective metabolic pathways may have a synergistic effect.
• Local cerebral blood flow assessment using transcranial Doppler ultrasonography in a dog with brain infarction in the right middle cerebral artery territory.

  Kazuyoshi Sasaoka, Hiroshi Ohta, Tomohito Ishizuka, Tatsuyuki Osuga, Keitaro Morishita, Noboru Sasaki, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 84 (10) 1385 - 1390 2022/10/01 [Refereed]
   

  A 12-year-old neutered male Chihuahua was diagnosed with acute brain infarction in the right middle cerebral artery (MCA) territory. Transcranial Doppler ultrasonography (TCD) was performed to assess the local cerebral blood flow at the time of diagnosis and after 4 and 31 hr. Initially, the right MCA retained blood flow but with a lower cerebral blood flow velocity (CBFV; 14.9 cm/sec) than the left MCA (27.9 cm/sec). The TCD vascular resistance variables were higher in the right than in the left MCA. An increase in the CBFV and a decrease in TCD vascular resistance variables were observed, consistent with improvements in neurological symptoms. TCD can be a non-invasive, and easy-to-use modality for bedside monitoring of cerebral edema and infarction.
• Malignant oligoastrocytoma in the spinal cord of a cat.

  Dai Hasegawa, Keisuke Aoshima, Kazuyoshi Sasaoka, Atsushi Kobayashi, Mitsuyoshi Takiguchi, Takashi Kimura

  The Journal of veterinary medical science 84 (9) 1277 - 1282 0916-7250 2022/09/12 [Refereed]
   

  A 12-year and 3-month spayed female mixed cat was presented with severe lumbar pain. Magnetic resonance imaging and postmortem examination revealed a swollen lesion in the spinal cord at L3 level. Histologic examination identified extensive neoplastic cell proliferation with massive necrosis in the tumor tissue. Two types of neoplastic cells were recognized. One type of neoplastic cells were large cells characterized by round to polygonal shape and abundant eosinophilic cytoplasm (referred to as "large cells"). The other neoplastic cells were small, densely proliferated, and had round to irregular shape and scant eosinophilic cytoplasm (referred to as "small cells"). Both types of cells were positive for oligodendrocyte transcription factor 2 and SRY-box transcription factor 10. Glial fibrillary acidic protein was positive in large cells but negative in most small cells. Digital analysis for Ki-67-stained tumor tissues found that total 21.1% ± 6.5% of tumor cells were positive for Ki-67. Based on these findings, we diagnosed malignant oligoastrocytoma in the spinal cord.
• Urinary corticoid to creatinine ratios using IMMULITE 2000 XPi for diagnosis of canine hypercortisolism.

  Noriyuki Nagata, Hiryu Sawamura, Keitaro Morishita, Kenji Hosoya, Nozomu Yokoyama, Kazuyoshi Sasaoka, Noboru Sasaki, Kensuke Nakamura, Yoshinori Ikenaka, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 84 (7) 954 - 959 2022/07/10 [Refereed]
   

  The urinary corticoid to creatinine ratio (UCCR) is one of the most commonly used screening tests for canine hypercortisolism (HC). In this study, a reference interval was established for UCCR using IMMULITE 2000 XPi, the latest chemiluminescence enzyme immunoassay. The diagnostic performance of this method for UCCR in canine HC was also evaluated. The median UCCR was 1.06 × 10-5 (range: 0.28-2.49) for 58 healthy dogs, and an upper reference limit of 1.98 × 10-5 (90% confidence interval: 1.76-2.15) was determined. The median UCCR in the 12 dogs with HC (7.38 × 10-5, range 1.86-29.98) was significantly higher than that in the 16 dogs with mimic-HC (1.59 × 10-5, range 0.47-3.42, P<0.001). The area under the curve for UCCR to differentiate HC dogs from mimic-HC dogs was 0.971, with a sensitivity of 91.7% and specificity of 100% when the cut-off value was set at 3.77 × 10-5. The UCCR of 16 paired urine samples collected at home and in hospital showed that the UCCR of samples collected in the hospital was significantly higher than that of samples collected at home (mean difference 3.30 × 10-5, 95% confidence interval: 0.70-5.90, P=0.001). In summary, we established the upper reference limit for UCCR using IMMULITE 2000 XPi in dogs and confirmed that UCCR is a useful diagnostic test for HC in dogs if urine samples are collected at home.
• Characterization of mucin gene expression and goblet cell proportion in inflammatory colorectal polyps in miniature dachshunds.

  Noriyuki Nagata, Hiroshi Ohta, Nozomu Yokoyama, Yong Bin Teoh, Noboru Sasaki, Kensuke Nakamura, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 84 (6) 872 - 876 2022/06/18 [Refereed]
   

  Hyperplastic goblet cells and abundant mucus are significant characteristics of inflammatory colorectal polyps (ICRPs) in miniature dachshunds. In this study, selected mucin gene expressions and goblet cell proportions were evaluated in miniature dachshunds with ICRPs and in healthy dogs. Mucin 2 (MUC2) gene expression was not significantly different among the groups, whereas mucin 5AC (MUC5AC) gene expression was significantly higher in the polypoid lesions than in healthy colonic mucosa. Although the percentage of goblet cells in the upper crypt regions did not significantly differ between the groups, that in the lower crypt regions was significantly decreased in polypoid lesions. In conclusion, increased MUC5AC gene expression and goblet cell proportion changes may be associated with the pathogenesis of ICRPs.
• Incremental predictive value of echocardiographic indices of right ventricular function in the assessment of long-term prognosis in dogs with myxomatous mitral valve disease

  T. Morita, K. Nakamura, T. Osuga, M. Takiguchi

  Journal of Veterinary Cardiology 39 51 - 62 1760-2734 2022/02 [Refereed]
   

  Introduction: Few studies have evaluated the utility of echocardiographic indices of right ventricular (RV) function in predicting prognosis in dogs with myxomatous mitral valve disease (MMVD). Animals: Sixty-seven client-owned dogs were diagnosed with MMVD. Materials and methods: clinical cohort study. Dogs diagnosed with American college of veterinary internal medicine (ACVIM) stage B2, C or D between April 2014 and March 2017 were enrolled. Long-term outcomes were assessed by telephone or from the medical record. The primary end-point was defined as cardiac-related death. Echocardiographic indices of RV function, including the RV Tei index, free wall and septal RV longitudinal strain, were obtained. Univariable and multivariable Cox proportional hazard analyses were used to identify variables predictive of cardiac-related death. Results: Twenty-four dogs died during the follow-up period. The median follow-up time was 482 days, and the median survival time for dogs with cardiac-related death was 230 days. For cardiac-related death, peak early diastolic mitral inflow velocity, ACVIM stage C or D, tricuspid regurgitation velocity, RV Tei index and RV end-diastolic area were predictors in univariable Cox proportional hazard analysis. In multivariable Cox proportional analysis adjusted for the left atrial to aorta ratio, peak early diastolic mitral inflow velocity and ACVIM stage, an increase in the Tei index by 0.1 increased the hazard ratio of cardiac-related death by 33% (95% confidence interval, 16–70%; P = 0.002). Conclusions: In dogs with MMVD, RV dysfunction assessed by the Tei index is an independent predictor of cardiac-related death.
• Transcranial Doppler ultrasonography detects the elevation of cerebral blood flow during ictal-phase of pentetrazol-induced seizures in dogs.

  Kazuyoshi Sasaoka, Hiroshi Ohta, Tomohito Ishizuka, Kazuki Kojima, Noboru Sasaki, Mitsuyoshi Takiguchi

  American journal of veterinary research 83 (4) 331 - 338 2022/01/28 [Refereed]
   

  OBJECTIVE: To investigate the association between changes in cerebral blood flow and electrographic epileptic seizure in dogs using transcranial Doppler ultrasonography (TCD). ANIMALS: 6 healthy Beagle dogs. PROCEDURES: Each dog was administered pentetrazol (1.5 mg/kg/min) or saline (0.9% NaCl) solution under general anesthesia with continuous infusion of propofol. Both pentetrazol and saline solution were administered to all 6 dogs, with at least 28 days interval between the experiments. Blood flow waveforms in the middle cerebral artery and the basilar artery were obtained using TCD at baseline, after pentetrazol administration, and after diazepam administration. TCD velocities, including peak systolic velocity, end-diastolic velocity, and mean velocity and resistance variables, were determined from the Doppler waveforms. RESULTS: During ictal-phase of pentetrazol-induced seizures, the TCD velocities significantly increased in the basilar and middle cerebral arteries while TCD vascular resistance variables did not change in either artery. The TCD velocities significantly decreased after diazepam administration. Systemic parameters, such as the heart rate, mean arterial pressure, systemic vascular resistance, cardiac index, end-tidal carbon dioxide, oxygen saturation, and body temperature, did not change significantly during seizures. CLINICAL RELEVANCE: This study showed that cerebral blood flow, as obtained from TCD velocities, increased by 130% during ictal-phase of pentetrazol-induced seizures in dogs. The elevated velocities returned to baseline after seizure suppression. Thus, TCD may be used to detect electrographic seizures during the treatment of status epilepticus in dogs, and further clinical studies clarifying the association between changes in cerebral blood flow and non-convulsive seizure cases are needed.
• Serum steroid profiling of hepatocellular carcinoma associated with hyperadrenocorticism in dogs: A preliminary study.

  Thandar Oo, Noboru Sasaki, Yoshinori Ikenaka, Takahiro Ichise, Noriyuki Nagata, Nozomu Yokoyama, Kazuyoshi Sasaoka, Keitaro Morishita, Kensuke Nakamura, Mitsuyoshi Takiguchi

  Frontiers in veterinary science 9 1014792 - 1014792 2022 [Refereed]
   

  BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common primary liver tumors in humans and dogs. Excessive adrenocortical hormone exposure may cause steroid hepatopathy, which may develop into HCC. In our previous study, hyperadrenocorticism (HAC) was a highly concurrent disease in dogs with HCC. Therefore, this study hypothesized that adrenal steroid alterations might be involved in hepatocarcinogenesis and aimed to specify the relationship between HAC and HCC in dogs. MATERIALS AND METHODS: This study included 46 dogs brought to the Hokkaido University Veterinary Teaching Hospital between March 2019 and December 2020. Owners gave their signed consent for blood collection on their first visit. A total of 19 steroids (14 steroids and 5 metabolites) in the baseline serum of 15 dogs with HCC, 15 dogs with HAC, and 10 dogs with both diseases were quantitatively measured using the developed liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS) method. RESULTS: In each group, 11 steroids were detected higher than 50%. The detection rate of steroid hormones did not significantly differ between the groups (p > 0.05). Principle component analysis (PCA) showed that the steroid profiles of the three groups were comparable. Median steroid hormone concentrations were not significantly different between the study diseases (p > 0.05). CONCLUSION: The developed LC/MS/MS was useful for measuring steroid hormones. Although it was clear that HAC was concurrent in dogs with HCC, none of the serum steroids was suggested to be involved in HCC.
• Evaluation of Right Ventricular Function and Dyssynchrony in a Dog Model of Acute Pulmonary Embolism: Diagnostic Utility and Reversibility.

  Tomoya Morita, Kensuke Nakamura, Tatsuyuki Osuga, Sei Kawamoto, Shingo Miki, Mitsuyoshi Takiguchi

  Frontiers in veterinary science 9 861064 - 861064 2022 [Refereed]
   

  Background: The diagnosis of acute pulmonary thromboembolism is challenging in dogs. Little has been reported on changes in echocardiographic indices in dogs with acute pulmonary thromboembolism. The objective of this study was to validate the relationship between echocardiographic indices and right heart catheterization variables in dogs with acute pulmonary embolism and to identify a useful echocardiographic index for diagnosing acute pulmonary embolism. Materials and Methods: Six healthy laboratory beagles were included in the study. Echocardiography and right heart catheterization were performed in a dog model of acute pulmonary embolism produced by microsphere injection. Echocardiographic indices, including the right ventricular (RV) Tei index, RV longitudinal strain, and the dyssynchrony index using speckle tracking echocardiography, transmitral flow, and eccentricity index, were measured. Results: The mean pulmonary arterial pressure increased (22.2 ± 1.2 mmHg) and the blood pressure decreased after microsphere injection. Although the mean pulmonary arterial pressure remained elevated, the blood pressure recovered 2 days after the microsphere injection. Most echocardiographic indices of RV function were significantly impaired following microsphere injection and recovered after 2 days. In contrast, the RV Tei index was significantly impaired after microsphere injection and the impairment persisted after 2 days. Multivariable analysis revealed that the RV Tei index was an independent echocardiographic predictor of pulmonary vascular resistance (β = 0.88, P < 0.001), and transmitral early diastolic wave was an independent predictor of the cardiac index (β = -0.86, P = 0.001). Conclusions: The RV Tei index is a useful echocardiographic index for diagnosing acute pulmonary embolism. Ventricular interdependence may be an important factor causing low cardiac output in dogs with acute pulmonary embolism.
• Case Report: Usefulness of Drip Infusion Cholangiography With Computed Tomography for the Diagnosis of Biloma in a Dog.

  Masahiro Tamura, Hiroshi Ohta, Nene Hasegawa, Kenji Hosoya, Mitsuyoshi Takiguchi

  Frontiers in veterinary science 9 854042 - 854042 2022 [Refereed]
   

  Bilomas are encapsulated collections of bile outside or inside the biliary tract within the abdominal cavity. For diagnostic and therapeutic approaches, it is important to identify the origin of bile leaks from the biliary tract. This case report describes the usefulness of drip infusion cholangiography with computed tomography (DIC-CT) for detecting the site of bile leakage in a dog with biloma. A 10-year-old, castrated male Pomeranian dog was referred to our department for gastrointestinal signs. Abdominal ultrasonography detected gallbladder mucocele without evidence of defect on the wall and well-defined anechoic localized fluid accumulation around the right division of the liver. On the other hand, there was only a small amount of ascites in the abdominal cavity. The accumulated fluid collected through abdominocentesis had a bilirubin concentration of 11.4 mg/dl, which was more than twice as high as that in serum (0.4 mg/dl), but had absence of pyogenic bacteria. The DIC-CT with meglumine iotroxate showed two well-defined large fluid collections: one between right medial and lateral lobe and the other between the right lateral lobe and caudate process of caudate lobe. Three-dimensional DIC-CT views that the former was enhanced by the contrast agent and that it communicated with an intrahepatic bile duct of the right lateral lobe. Moreover, the DIC-CT images confirmed communication with each fluid collections. After 6 days of hospitalization, a decrease in the amount of accumulated fluid was confirmed, after which cholecystectomy was performed. The dog was discharged from the hospital without complications. No signs of bile leakage were observed on follow-up imaging on postoperative day 10. According to authors knowledge, this has been the first report to show that DIC-CT can be useful for determining the origin of bile leakage in dogs with bilomas.
• Safety Assessment of Ultrasound-Assisted Intravesical Chemotherapy in Normal Dogs: A Pilot Study.

  Noboru Sasaki, Yoshinori Ikenaka, Keisuke Aoshima, Teiichiro Aoyagi, Nobuki Kudo, Kensuke Nakamura, Mitsuyoshi Takiguchi

  Frontiers in pharmacology 13 837754 - 837754 2022 [Refereed]
   

  Intravesical chemotherapy after transurethral resection is a treatment option in patients with non-muscle invasive bladder cancer. The efficacy of intravesical chemotherapy is determined by the cellular uptake of intravesical drugs. Therefore, drug delivery technologies in the urinary bladder are promising tools for enhancing the efficacy of intravesical chemotherapy. Ultrasound-triggered microbubble cavitation may enhance the permeability of the urothelium, and thus may have potential as a drug delivery technology in the urinary bladder. Meanwhile, the enhanced permeability may increase systemic absorption of intravesical drugs, which may increase the adverse effects of the drug. The aim of this preliminary safety study was to assess the systemic absorption of an intravesical drug that was delivered by ultrasound-triggered microbubble cavitation in the urinary bladder of normal dogs. Pirarubicin, a derivative of doxorubicin, and an ultrasound contrast agent (Sonazoid) microbubbles were administered in the urinary bladder. Ultrasound (transmitting frequency 5 MHz; pulse duration 0.44 μsec; pulse repetition frequency 7.7 kHz; peak negative pressure -1.2 MPa) was exposed to the bladder using a diagnostic ultrasound probe (PLT-704SBT). The combination of ultrasound and microbubbles did not increase the plasma concentration of intravesical pirarubicin. In addition, hematoxylin and eosin staining showed that the combination of ultrasound and microbubble did not cause observable damages to the urothelium. Tissue pirarubicin concentration in the sonicated region was higher than that of the non-sonicated region in two of three dogs. The results of this pilot study demonstrate the safety of the combination of intravesical pirarubicin and ultrasound-triggered microbubble cavitation, that is, ultrasound-assisted intravesical chemotherapy.
• Prognostic value of left ventricular-arterial coupling estimated using echocardiography in dogs with myxomatous mitral valve disease.

  Tatsuyuki Osuga, Tomoya Morita, Noboru Sasaki, Keitaro Morishita, Hiroshi Ohta, Mitsuyoshi Takiguchi

  Journal of veterinary internal medicine 35 (6) 2607 - 2615 2021/11/06 [Refereed]
   

  BACKGROUND: The interaction between the left ventricle (LV) and systemic arterial systems, known as left ventricular-arterial coupling (VAC), has been evaluated based on the effective arterial elastance (Ea) to LV end-systolic elastance (Ees) ratio (Ea/Ees). The Ea reflects the total arterial load of LV, whereas Ees reflects the LV systolic function. A recent study found that inappropriate VAC based on increased Ea/Ees estimated by echocardiography is associated with advanced disease severity in dogs with myxomatous mitral valve disease (MMVD). HYPOTHESIS: Inappropriate VAC assessed by echocardiographic estimation of Ea/Ees is associated with a worse prognosis in dogs with MMVD. ANIMALS: Eighty-nine dogs with MMVD. METHODS: Prospective cohort study. Dogs underwent echocardiographic examinations at enrollment. The Ea was estimated using the formula: mean blood pressure/(forward stroke volume/body weight). The Ees was estimated using the formula: mean blood pressure/(LV end-systolic volume/body weight). The Ea/Ees was calculated. RESULTS: By end of study, 22 dogs died of cardiac-related causes with 67 dogs censored. Dogs with increased Ea/Ees (Ea/Ees >0.34; median survival time, 527 days; 95% confidence interval [CI], 322 days-not determinable) had a shorter survival time (P < .0001) than those without increased Ea/Ees (Ea/Ees ≤0.34; median survival time, >1112 days; 95% CI, not determinable). Multivariate Cox proportional hazard analysis showed that Ea/Ees, body weight, peak systolic mitral annular velocity, and the peak early diastolic transmitral velocity-to-peak early diastolic mitral annular velocity ratio were independent predictors of cardiac-related death among echocardiographic indices. CONCLUSIONS AND CLINICAL IMPORTANCE: Inappropriate VAC assessed based on echocardiographically-estimated Ea/Ees is associated with a worse prognosis in dogs with MMVD.
• The use of 4-dimensional magnetic resonance angiography as a noninvasive procedure to diagnose subclavian steal syndrome in a dog.

  Genya Shimbo, Ryosuke Echigo, Mitsuyoshi Takiguchi

  Journal of the American Veterinary Medical Association 259 (9) 1040 - 1042 2021/11/01 [Refereed]
   

  CASE DESCRIPTION: A 5-year-old 4.1-kg (9.0-lb) spayed female Toy Poodle was referred because of a 6-month history of sporadic signs of neck pain. CLINICAL FINDINGS: Diagnostic imaging with MRI and CT revealed a dilated radicular artery connecting the right and left vertebral arteries and causing mild compression of the spinal cord. The left subclavian artery caudal to the origin of the left vertebral artery was absent. Subclavian steal syndrome (SSS) was suspected. TREATMENT AND OUTCOME: The owner declined surgical treatment; thus, the dog was treated conservatively with glucocorticoids and analgesics. Eight months later, the dog's clinical signs were unchanged but palliated with the administration of glucocorticoids and analgesics, and 4-dimensional (4-D) magnetic resonance angiography (MRA) revealed that the left vertebral artery received blood supply from the right vertebral artery through the dilated radicular artery and that the left vertebral artery caudal to this site had retrograde flow and drained into the left subclavian artery, confirming the diagnosis of SSS. The owner again declined surgery, and conservative treatment continued. The dog's condition was unchanged at the last follow-up communication 11 months after 4-D MRA. CLINICAL RELEVANCE: Subclavian steal syndrome is an extremely rare condition in dogs, and our findings suggested that 4-D MRA could be used to definitively diagnose SSS in dogs.
• Effects of intravenous administration of pimobendan on hemodynamic indices and indices of left atrial longitudinal strain derived from speckle-tracking echocardiography in healthy dogs.

  Shingo Miki, Kensuke Nakamura, Tomoya Morita, Tatsuyuki Osuga, Sei Kawamoto, Noboru Sasaki, Mitsuyoshi Takiguchi

  American journal of veterinary research 82 (10) 795 - 801 2021/10 [Refereed]
   

  OBJECTIVE: To determine the effects of IV administration of pimobendan on hemodynamic indices and indices of left atrial (LA) longitudinal strain by speckle-tracking echocardiography in healthy dogs. ANIMALS: 6 healthy Beagles. PROCEDURES: After the dogs were anesthetized, the right heart was catheterized and 2-D conventional and speckle-tracking echocardiography were performed before and after IV administration of 0.15 mg of pimobendan/kg. Speckle-tracking echocardiography was performed to assess the 3 LA phasic functions through LA deformation (longitudinal strain and strain rate) and volumetric analyses. RESULTS: Pimobendan significantly increased stroke volume and cardiac output and decreased systemic vascular resistance. Pimobendan significantly improved left ventricular function assessed by the Tei index and LA booster pump function assessed by LA longitudinal strain and change in fractional volume during atrial systole. Indices of LA reservoir and conduit function were unchanged. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy dogs, IV administration of pimobendan improved the indices of LA booster pump function but had no effect on the indices of LA reservoir and conduit functions. Further studies are needed to validate whether these results can be extrapolated to dogs with myxomatous mitral valve disease.
• Genome-wide DNA methylation analysis of dogs with high lead exposure living near a lead mining area in Kabwe, Zambia.

  Jumpei Yamazaki, Haruya Toyomaki, Shouta M M Nakayama, John Yabe, Kaampwe Muzandu, Jaroslav Jelinek, Shoko Yokoyama, Yoshinori Ikenaka, Mitsuyoshi Takiguchi, Mayumi Ishizuka

  Environmental pollution (Barking, Essex : 1987) 286 117229 - 117229 2021/10/01 [Refereed]
   

  Lead (Pb) is a heavy metal that has been proven to be toxic to both animals and humans. Genom-wide DNA methylation in domestic dogs exposed to high levels of Pb in Kabwe, Zambia was analyzed in this study. Using next-generation sequencing on samples from 20 domestic dogs (mean blood Pb concentration: 43.6 μg/dL and 7.2 μg/dL in the high and low exposure groups), a digital restriction enzyme analysis of methylation was performed to identify the genomic locations of differentially methylated CpG sites. A validation study on an additional 20 dogs followed (blood Pb concentration: 4.9-29.7 μg/dL). The cluster analysis resolved two broad clusters indicating high and low Pb exposure. The study identified 827 (1.2%) CpG sites with differences in methylation (101 CpG sites were hypermethylated in the low exposure group and 726 were hypermethylated in the high exposure group). The sites corresponded to 26 genes with differentially methylated CpG sites at their promoter regions, including the NGF gene. The methylation of four CpG sites was validated using bisulfite pyrosequencing. The results indicate that aberrant hypermethylation is prevalent in dogs exposed to Pb. The altered DNA methylation of the genes identified in this study contributes to a greater understanding of the epigenetic changes caused by Pb exposure and highlights novel biomarker discoveries across species.
• Obese status is associated with accelerated DNA methylation change in peripheral blood of senior dogs.

  Jumpei Yamazaki, Shinji Meagawa, Jaroslav Jelinek, Shoko Yokoyama, Noriyuki Nagata, Masashi Yuki, Mitsuyoshi Takiguchi

  Research in veterinary science 139 193 - 199 2021/10 [Refereed]
   

  Obesity and its associated comorbidities constitute a major and growing health problem worldwide not only involved with people but also dogs and cats. Although few genetic mutations have been associated with obesity in dogs, molecular mechanism remains to be clearly understood. Given the fact that DNA methylation leads to gene expression variability and has plasticity affected by metabolic phenotypes such as obesity in human, the objective of this study is to identify obesity-associated differentially methylated cytosine-phosphate-guanine (CpG) dinucleotide sites in dogs. With genome-wide DNA methylation analysis using next-generation sequencing for blood samples from fourteen Miniature dachshunds with body condition score (BCS) 4-5 and BCS ≥6, over 100,000 sites could be analysed to identify genomic locations of differentially methylated CpG sites. As a result, 191 differentially methylated CpG sites (89 CpG sites were hypermethylated in BCS ≥6 and 102 were hypermethylated in BCS 4-5) were identified. These sites included promoter regions of Kisspeptin receptor (KISS1R) and Calcyphosine 2 (CAPS2) genes which were subsequently validated by bisulfite-pyrosequencing for another set of 157 dog blood samples. KISS1R methylation levels were found to be higher in BCS ≥6 group than BCS 4-5 in senior (>84 months) dogs. Especially male dogs but not female dogs as well as uncastrated male dogs but not castrated male dogs showed this trend. DNA methylation of KISS1R gene will be useful for understanding of comprehensive epigenetic change in obese dogs.
• Simultaneous steroids measurement in dogs with hyperadrenocorticism using a column-switching liquid chromatography-tandem mass spectrometry method.

  Noboru Sasaki, Thandar Oo, Yoshikazu Yasuda, Takahiro Ichise, Noriyuki Nagata, Nozomu Yokoyama, Kazuyoshi Sasaoka, Keitaro Morishita, Shouta Mm Nakayama, Mayumi Ishizuka, Kensuke Nakamura, Mitsuyoshi Takiguchi, Yoshinori Ikenaka

  The Journal of veterinary medical science 83 (11) 1634 - 1642 2021/09/17 [Refereed]
   

  We developed an analytical method using an on-line column-switching liquid chromatography with triple quadrupole mass spectrometry (LC/MS/MS) for quantifying multiple steroids in serum. Using the developed method, we evaluated the serum concentration of nine steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 21-deoxycortisol, deoxycorticosterone, progesterone, 17α-OH-progesterone and aldosterone) in dogs with hyperadrenocorticism (HAC). Serum was mixed with stable isotope internal standards and thereafter purified by the automated column-switching system. The limit of detection ranged 2-16 pg/ml for nine steroids. In the baseline samples, five steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, and 17α-OH-progesterone) were detected in all dogs. The concentrations of cortisone, 11-deoxycortisol, and 17α-OH-progesterone in dogs with HAC (n=19) were significantly higher those in dogs without HAC (n=15, P<0.02). After the adrenocorticotropic hormone stimulation test, six steroids (cortisol, corticosterone, cortisone, 11-deoxycortisol, 17α-OH-progesterone, and deoxycorticosterone) were above the limit of quantification in all dogs. Cortisol, corticosterone, cortisone, and deoxycorticosterone concentrations of dogs with HAC were significantly higher than those of dogs without HAC (P<0.02). In addition, 11-deoxycortisol and 17α-OH-progesterone concentration was higher in dogs with HAC than in dogs without HAC (P=0.044 and P=0.048, respectively). The on-line column-switching LC/MS/MS would be feasible for measuring multiple steroids in dog serum. The results suggest that cortisone, 11-deoxycortisol, and 17α-OH-progesterone would be related to HAC. Further studies are warranted to assess the clinical feasibility of steroid profile in dogs with HAC.
• Genome-wide DNA methylation profile in feline haematological tumours: A preliminary study.

  Jumpei Yamazaki, Jaroslav Jelinek, Shoko Yokoyama, Mitsuyoshi Takiguchi

  Research in veterinary science 140 221 - 228 2021/09/08 [Refereed]
   

  Although DNA methylation has been analysed in few studies for a limited number of loci in cats with diseases, genome-wide profile of DNA methylation has never been addressed. The hypothesis for this study is that next-generation sequencing with sequential digestion of genomic DNA with SmaI and XmaI enzymes could provide highly quantitative information on methylation levels in cats. Using blood from four healthy control cats and two disease cats as well as three feline lymphoma/leukemia cell lines, approximately 74-94 thousand CpG sites across the cat genome could be analysed. CpG sites in CpG island (CGI) were broadly either methylated or unmethylated in normal blood, while CpG sites in non-CpG islands (NCGI) are largely methylated. Lymphoma cell lines showed thousands of CpG sites with gain of methylation at normally unmethylated CGI sites and loss of methylation at normally methylated NCGI sites. Hypermethylated CpG sites located at promoter regions included genes annotated with 'developmental process' and 'anatomical structure morphogenesis' such as HOXD10. This highly quantitative method would be suitable for studies of DNA methylation changes not only in cancer but also in other common diseases in cats.
• Systemic mucoid degeneration of the arterial tunica intima in a young dog.

  Yoshiki Ichikawa, Mizuki Heishima, Kristin Vyhnal, Keisuke Aoshima, Kazuyoshi Sasaoka, Ryohei Kinoshita, Atsushi Kobayashi, Mitsuyoshi Takiguchi, Takashi Kimura

  Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 34 (1) 10406387211042587 - 10406387211042587 2021/09/04 [Refereed]
   

  A 27-mo-old, spayed female mixed-breed dog was presented with left forelimb pain, which progressed to full thickness necrosis of the soft tissues of multiple limbs. Clinical imaging and postmortem examination suggested multiple large arterial thromboemboli. Histologic examination of vascular lesions revealed markedly thickened tunica intima with polypoid intraluminal projections, which partially to entirely occluded the arterial lumen. The expanded tunica intima was comprised of intimal accumulation of Alcian blue-positive matrix with scattered spindle-to-satellite cells. These cells were positive for von Willebrand factor and vimentin but negative for α-smooth muscle actin, suggesting endothelial origin. Deposition of the intimal mucoid matrix was observed in the elastic and muscular arteries associated with regional ischemic changes. Mucoid emboli, likely from fragmentation of proliferative intimal tissue, were identified in smaller vessels supplied by affected arteries. Based on these findings, we diagnosed systemic mucoid degeneration of the arterial tunica intima. Such systemic arterial degeneration characterized by deposition of mucoid matrix in the tunica intima has not been reported previously in dogs, to our knowledge, and should be distinguished from thromboembolism and other degenerative vascular diseases.
• Cardiovascular effects of dose escalating of norepinephrine in healthy dogs anesthetized with isoflurane.

  Kazuki Kojima, Tomohito Ishizuka, Noboru Sasaki, Kensuke Nakamura, Mitsuyoshi Takiguchi

  Veterinary anaesthesia and analgesia 48 (5) 654 - 662 2021/09 [Refereed]
   

  OBJECTIVE: To evaluate the systemic cardiovascular effects of dose escalating administration of norepinephrine in healthy dogs anesthetized with isoflurane. STUDY DESIGN: Experimental study. ANIMALS: A total of six adult laboratory Beagle dogs, 10.5 (9.2-12.0) kg [median (range)]. METHODS: Each dog was anesthetized with isoflurane at an end-tidal concentration of 1.7%, mechanically ventilated and administered a continuous rate infusion of rocuronium (0.5 mg kg-1 hour-1). Each dog was administered incremental dose rates of norepinephrine (0.05, 0.125, 0.25, 0.5, 1.0 and 2.0 μg kg-1 minute-1), and each dose was infused for 15 minutes. Cardiovascular variables were recorded before administration and at the end of each infusion period. RESULTS: Norepinephrine infusion increased mean arterial pressure (MAP), cardiac output (CO) and oxygen delivery in a dose-dependent manner. Systemic vascular resistance did not significantly change during the experiment. Stroke volume increased at the lower dose rates and heart rate increased at the higher dose rates. Oxygen consumption and lactate concentrations did not significantly change during infusions. CONCLUSIONS: In dogs anesthetized with isoflurane, norepinephrine increased MAP by increasing the CO. CO increased with a change in stroke volume at lower dose rates of norepinephrine. At higher dosage, heart rate also contributed to an increase in CO. Norepinephrine did not cause excessive vasoconstriction that interfered with the CO during this study. CLINICAL RELEVANCE: Norepinephrine can be useful for treating hypotension in dogs anesthetized with isoflurane.
• Peliosis Hepatis with Chylous Ascites in a Dog.

  Kevin Christian Montecillo Gulay, Noriyuki Nagata, Keisuke Aoshima, Nozomi Shiohara, Atsushi Kobayashi, Mitsuyoshi Takiguchi, Takashi Kimura

  Journal of comparative pathology 187 63 - 67 2021/08 [Refereed]
   

  A 6-year-old spayed female Toy Poodle dog was referred to the Hokkaido University Veterinary Teaching Hospital for abdominal distension. Abdominocentesis yielded ascitic fluid that had a mildly increased total protein concentration and a 2.7-fold higher triglyceride concentration than plasma, and was interpreted as chylous ascites. The patient had an enlarged liver, which contained multiple, small, nodular masses and cyst-like structures. Microscopically, these lesions were multifocal dilated spaces containing lymphocytes, endothelial cells, fibrin and islands of hepatocytes. Increased α-smooth muscle actin-positive cells were observed in hepatic sinusoids. Based on these findings, we diagnosed peliosis hepatis with chylous ascites, which is likely to have been due to lymphangiectasia and disrupted hepatic sinusoids. Neither Bartonella spp DNA nor mutations in ACVRL1 and MTM1 genes were detected, although there was a 47-fold increase in hepatic ACVRL1 expression compared with age-matched control liver. To the authors' knowledge, this is the first report of chylous ascites resulting from peliosis hepatis in any species.
• CT morphology of anomalous systemic arterial supply to normal lung in dogs.

  Genya Shimbo, Mitsuyoshi Takiguchi

  Veterinary radiology & ultrasound : the official journal of the American College of Veterinary Radiology and the International Veterinary Radiology Association 62 (6) 657 - 665 2021/07/11 [Refereed]
   

  Anomalous systemic arterial supply to the normal lung (ASANL) is a rare congenital anomaly in humans, in which the systemic arteries supply the basal segments of the lower lobe. It has a normal bronchial connection, but lacks a normal pulmonary artery. This anomaly has not been previously reported in the veterinary literature. The objectives of this retrospective descriptive study were to characterize the CT findings and clinical features of ASANL , and to determine the breed predisposition in a population of referral canine cases. Thoracic CT images, in which the caudal lung lobes were fully inflated and the pulmonary artery could be traced to the periphery, were reviewed. A total of 1,950 dogs were enrolled, and the aberrant vasculature equivalent to ASANL in humans was detected in 48 dogs. Shetland Sheepdogs (7/48, odds ratio [OR] = 8.0, P < 0.00001), Miniature Dachshunds (19/48, OR = 3.9, P < 0.00001), and Labrador Retrievers (6/48, OR = 4.5, P = 0.0009) were over-represented. The affected lung lobes were the right caudal lobe (24/48, 50%), the left caudal lobe (21/48, 43.8%), and bilateral caudal lobes (3/48, 6.3%). The aberrant vessels originated from the left gastric artery (14/48), descending thoracic aorta (8/48), celiac artery (6/48), and splenic artery (1/48). In the remaining 19 cases, the origin of the aberrant vessels could not be determined. Although the clinical significance of ASANL in dogs remains unclear, surgeons should be aware of this finding prior to lobectomy of the caudal lung lobes to avoid intraoperative systemic arterial bleeding.
• Morphometric evaluation of canine hepatocellular carcinoma using computed tomography: a promising tool for predicting malignancy.

  Rommaneeya Leela-Arporn, Hiroshi Ohta, Genya Shimbo, Noboru Sasaki, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 83 (9) 1459 - 1464 2021/07/06 [Refereed]
   

  The size of canine focal liver lesions (FLLs) is known to be one of the predicting criteria for malignancy. However, there are discrepancies for the measurement of maximum lesion size, resulting in contradicting results among studies and incidences of false positive outcomes. Thus far, the morphometric changes of FLLs for distinguishing malignancy from benignancy remains undocumented. This study aimed to investigate morphometric characteristics of FLLs using computed tomography (CT). CT images of 40 dogs with histopathological confirmation of 49 liver lesions, including 39 hepatocellular carcinomas and 10 nodular hyperplasias were retrospectively reviewed. The morphometric parameters including size (long and short axis diameters measured on transverse image), shape (measured by long to short axis (L/S) ratio), volume, and surface appearance of a liver lesion were evaluated using univariate and stepwise multivariate analyses, respectively. The results of univariate analysis showed that long and short axis diameters, L/S ratio, volume, and surface appearance of a lesion were significantly different between hepatocellular carcinomas and nodular hyperplasias. Multivariate analysis revealed that short axis diameter (>3.30 cm; odds ratio (OR): 36.1, 95% confidence interval (CI): 3.36-387.05, P=0.0031) and L/S ratio (>1.23; OR: 18.1, 95% CI: 1.61-205.12, P=0.0191) were independent predictors of malignancy, with the area under the curve of 0.9154. These results suggest that the combination of short axis diameter and L/S ratio is a promising tool for predicting liver malignancy with outstanding discriminating ability.
• Extrahepatic biliary obstruction can interfere with hepatic fibrosis prediction using two-dimensional shear wave elastography in dogs.

  Masahiro Tamura, Hiroshi Ohta, Tatsuyuki Osuga, Noboru Sasaki, Keitaro Morishita, Mitsuyoshi Takiguchi

  Veterinary radiology & ultrasound : the official journal of the American College of Veterinary Radiology and the International Veterinary Radiology Association 62 (4) 483 - 489 2021/07 [Refereed]
   

  Two-dimensional shear wave elastography (2D-SWE) can be used to quantitatively evaluate the elastic modulus of the liver as shear wave velocity (SWV), which can noninvasively predict clinically relevant hepatic fibrosis in both dogs and humans. However, extrahepatic biliary obstruction (EHBO), regardless of the presence of clinically relevant hepatic fibrosis, can influence SWVs in humans and thus may interfere with hepatic fibrosis prediction using 2D-SWE in dogs. The aim of this prospective, observational, and one-group pretest-posttest study is to investigate whether SWV measured by 2D-SWE displays a difference between dogs with and without EHBO. A total of 20 dogs were included (7 with EHBO and 13 with gallbladder pathology but no EHBO) that underwent preoperative SWV measurement using 2D-SWE. In all dogs, stages of hepatic fibrosis were evaluated histopathologically using a scoring scheme. In addition, postoperative SWVs in dogs with EHBO relieved via laparotomy were also evaluated. The median (range) SWVs in the dogs with and without EHBO were 1.91 (1.81-2.54) m/s and 1.57 (1.37-1.64) m/s, respectively. Although there was no significant difference in the histopathological hepatic fibrosis stages between the dogs with and without EHBO, the preoperative SWVs in the dogs with EHBO were significantly higher than in dogs without EHBO (P = .0004), and SWVs were found to decrease significantly after surgery (P = .0097). This study demonstrates that EHBO can increase the SWV of dogs without clinically relevant hepatic fibrosis and can interfere with the prediction of noninvasive hepatic fibrosis using 2D-SWE.
• The effects of fipronil on emotional and cognitive behaviors in mammals.

  Tomohiro Suzuki, Anri Hirai, Kraisiri Khidkhan, Collins Nimako, Takahiro Ichise, Kazuki Takeda, Hazuki Mizukawa, Shouta M M Nakayama, Kei Nomiyama, Nobuhiko Hoshi, Mizuki Maeda, Tetsushi Hirano, Kazuyoshi Sasaoka, Noboru Sasaki, Mitsuyoshi Takiguchi, Mayumi Ishizuka, Yoshinori Ikenaka

  Pesticide biochemistry and physiology 175 104847 - 104847 2021/06 [Refereed]
   

  Fipronil is a phenylpyrazole insecticide that is widely used as a pesticide and a veterinary drug, although studies suggest that it could be toxic to mammals. The objectives of this study were to examine the pharmacokinetic profile of fipronil in mice, dogs, and cats, and to evaluate its effects on emotional and cognitive behaviors of dogs and cats using the data obtained from mice. The assessment of in vivo kinetics of fipronil was conducted in mice and dogs. We also performed behavioral tests (elevated plus-maze and Y-maze) and measured the levels of neurotransmitters in mice exposed to fipronil. In addition, the in vitro metabolism of fipronil were evaluated using liver microsomes of rats, mice, dogs, and cats. The results revealed that fipronil is distributed throughout the body (blood, brain, adipose tissue, and liver) of mice after dermal application. It was metabolized to fipronil sulfone primarily in the liver. The data on kinetics show that both fipronil and fipronil sulfone have a longer half-life in dogs and cats than in mice. The behavioral tests indicated that fipronil and fipronil sulfone could affect emotional and cognitive behaviors and alter the levels of neurotransmitters (dopamine in the striatum and serotonin in the hippocampus) in mice. Furthermore, we found that dogs and cats have a low ability to metabolize fipronil than mice and rats. However, further comprehensive studies are needed to determine whether fipronil affects the emotional and cognitive behaviors when administered to dogs and cats. To the best of our knowledge, this is the first study to examine the pharmacokinetic data and verify the effects of fipronil on emotional and cognitive behaviors of dogs and cats using the data obtained from mice.
• DNA methylation landscape of 16 canine somatic tissues by methylation-sensitive restriction enzyme-based next generation sequencing.

  Jumpei Yamazaki, Yuki Matsumoto, Jaroslav Jelinek, Teita Ishizaki, Shingo Maeda, Kei Watanabe, Genki Ishihara, Junya Yamagishi, Mitsuyoshi Takiguchi

  Scientific reports 11 (1) 10005 - 10005 2021/05/11 [Refereed]
   

  DNA methylation plays important functions in gene expression regulation that is involved in individual development and various diseases. DNA methylation has been well studied in human and model organisms, but only limited data exist in companion animals like dog. Using methylation-sensitive restriction enzyme-based next generation sequencing (Canine DREAM), we obtained canine DNA methylation maps of 16 somatic tissues from two dogs. In total, we evaluated 130,861 CpG sites. The majority of CpG sites were either highly methylated (> 70%, 52.5-64.6% of all CpG sites analyzed) or unmethylated (< 30%, 22.5-28.0% of all CpG sites analyzed) which are methylation patterns similar to other species. The overall methylation status of CpG sites across the 32 methylomes were remarkably similar. However, the tissue types were clearly defined by principle component analysis and hierarchical clustering analysis with DNA methylome. We found 6416 CpG sites located closely at promoter region of genes and inverse correlation between DNA methylation and gene expression of these genes. Our study provides basic dataset for DNA methylation profiles in dogs.
• Urinary free metanephrines measurement in dogs with adrenal gland diseases using a new simple liquid chromatography tandem mass spectrometry method.

  Noboru Sasaki, Yoshinori Ikenaka, Yumiko Inoue, Takahiro Ichise, Noriyuki Nagata, Mayumi Ishizuka, Syouta Mm Nakayama, Kensuke Nakamura, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 83 (4) 648 - 655 2021/04/09 [Refereed]
   

  Measurement of urinary metanephrines in spot samples is used for the diagnosis of canine pheochromocytoma (PC). We describe a simple analytical method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for measuring free metanephrine (MN) and normetanephrine (NMN) in spot urine samples. Using the developed method, we evaluated the stability of urinary free-MN and free-NMN at various storing conditions. In addition, we assessed the feasibility of urinary free-MN and -NMN measurement for diagnosing PC. Urine samples were mixed with stable isotope internal standards and thereafter purified by ultrafiltration. The purified samples were analyzed by LC-MS/MS in the multiple reaction monitoring mode after separation on a multimode octa decyl silyl column. The coefficient of variation of free-MN and -NMN measurement was 7.6% and 5.5%, respectively. The linearity range was 0.5-10 µg/l for both analytes. Degradation was less than 10% for both analytes under any of the storage conditions. The median free-NMN ratio to creatinine of 9 PC dogs (595, range 144-47,961) was significantly higher (P<0.05) than that of 13 dogs with hypercortisolism (125, range 52-224) or 15 healthy dogs (85, range 50-117). The developed method is simple and may not require acidification of spot urine. The results of this preliminary retrospective study suggest that the measurement of urinary free metanephrines is a promising tool for diagnosing canine PC.
• Repeatability, reproducibility, and reference intervals for indices of right atrial longitudinal strain derived from speckle-tracking echocardiography in healthy dogs.

  Tomoya Morita, Kensuke Nakamura, Tatsuyuki Osuga, Mitsuyoshi Takiguchi

  American journal of veterinary research 82 (4) 274 - 279 2021/04 [Refereed]
   

  OBJECTIVE: To determine repeatability, reproducibility, and reference intervals of indices of right atrial longitudinal strain (RALS) derived from speckle-tracking echocardiography (STE) in dogs without heart disease. ANIMALS: 110 client-owned dogs and 10 laboratory Beagles. PROCEDURES: To determine intraobserver within-day (repeatability) and interobserver (reproducibility) coefficients of variation, RALS during ventricular systole (ϵS), ventricular early diastole (ϵE), and atrial systole (ϵA), as derived with STE, were obtained by 2 investigators for 5 randomly selected client-owned dogs and analyzed by linear regression. Reference intervals were estimated from the results of all dogs. Correlations between RALS indices (ϵS, ϵE, and ϵA) and sex, age, body weight, heart rate, and blood pressure were determined. RESULTS: RALS derived from STE showed good intraobserver within-day repeatability and interobserver reproducibility, with coefficients of variation of < 20%. Both ϵS and ϵE were significantly negatively correlated with age, but ϵA was not correlated with age. Indices were not correlated with sex, body weight, or blood pressure. CONCLUSIONS AND CLINICAL RELEVANCE: RALS indices derived from STE were repeatable and reproducible and were affected by the age of dogs without heart disease. Age should be considered in the interpretation of RALS indices in clinical settings. Further studies are needed to apply RALS indices for assessing dogs with heart disease.
• Prognostic value of small intestinal dilatation in dogs with protein-losing enteropathy.

  Hiroshi Ohta, Noriyuki Nagata, Nozomu Yokoyama, Tatsuyuki Osuga, Noboru Sasaki, Keitaro Morishita, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 83 (3) 378 - 384 2021/03/11 [Refereed]
   

  To date, little is known about the prognostic significance of ultrasonographic findings in dogs with protein-losing enteropathy (PLE). The aim of this retrospective study was to examine the prognostic value of ultrasonographic findings in dogs with PLE. A total of 26 dogs with PLE were included: 20 dogs with chronic enteropathy and 6 dogs with gastrointestinal lymphoma. The presence of small intestinal dilatation was associated with shorter survival time in dogs with PLE (P=0.003). The presence of hyperechoic intestinal mucosal striations was associated with longer survival time in dogs with PLE (P=0.0085). The results of the current study indicate that the presence of small intestinal dilatation might be associated with poor prognosis in dogs with PLE.
• Reduced expression levels of heat shock protein 90 in a diminazene aceturate-resistant Babesia gibsoni isolate.

  Masahiro Yamasaki, Natsuki Idaka, Moeko Abe, Mitsuyoshi Takiguchi

  Experimental parasitology 221 108050 - 108050 2021/02 [Refereed]
   

  Heat shock protein 90 (HSP90) is a molecular chaperon and an essential component for stage differentiation and intracellular growth inside the host cells of many protozoans. HSP90 of Babesia gibsoni (BgHSP90) was suggested to function in the development of diminazene aceturate (DA)-resistance. Therefore, we examined the expression level of BgHSP90 in a DA-resistant B. gibsoni isolate. Transcription of the BgHSP90 gene in the DA-resistant isolate and wild-type B. gibsoni was assessed by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). As a result, the copy number and relative amount of BgHSP90 transcripts in the DA-resistant isolate were significantly lower than those in the wild-type. Moreover, a rabbit anti-recombinant BgHSP90 antibody was developed, and the protein synthesis of BgHSP90 in the DA-resistant isolate was compared with that in the wild-type by Western blot analysis and indirect fluorescence assay. There was significantly less BgHSP90 protein than in the wild-type. Additionally, the relative intensity of BgHSP70 in DA-resistant isolate was also lower than that in the wild-type. This suggested that the expression of BgHSP90 and BgHSP70 in the DA-resistant B. gibsoni isolate was suppressed and that the reduced amount of BgHSP90 and BgHSP70 might cause the weak proliferation of the DA-resistant isolate. Further studies are necessary to elucidate the function of BgHSP90.
• Echocardiographic estimation of left ventricular-arterial coupling in dogs with myxomatous mitral valve disease.

  Tatsuyuki Osuga, Tomoya Morita, Noboru Sasaki, Keitaro Morishita, Hiroshi Ohta, Mitsuyoshi Takiguchi

  Journal of veterinary internal medicine 35 (1) 78 - 87 2021/01 [Refereed]
   

  BACKGROUND: The effective arterial elastance (Ea) to left ventricular (LV) end-systolic elastance (Ees) ratio (Ea/Ees) is an index of the interaction between LV and systemic arterial systems, left ventricular-arterial coupling (VAC). The Ea is an index of total arterial load of the LV, whereas Ees is an index of LV systolic function. In humans, inappropriate VAC based on increased Ea/Ees estimated using echocardiography is associated with more advanced heart disease severity. HYPOTHESIS: Left ventricular-arterial coupling assessed by echocardiographic estimation of Ea/Ees is associated with disease severity in dogs with myxomatous mitral valve disease (MMVD). ANIMALS: Ninety MMVD dogs and 61 healthy dogs. METHODS: Prospective cross-sectional study. The MMVD dogs were classified into stages B1, B2, or C according to American College of Veterinary Internal Medicine guidelines. Effective arterial elastance was echocardiographically estimated using the formula: mean blood pressure/(forward stroke volume/body weight). End-systolic elastance was echocardiographically estimated using the formula: mean blood pressure/(LV end-systolic volume/body weight). The ratio Ea/Ees was calculated. RESULTS: The ratio Ea/Ees was higher in stage B2 dogs than in healthy dogs and dogs stage B1 (both P < .0001), and higher in stage C dogs than in healthy dogs and dogs in the other 2 stages (healthy vs C and B1 vs C, P < .0001; B2 vs C, P = .0005). Multivariable logistic regression analysis showed that Ea/Ees and the peak velocity of early diastolic transmitral flow to isovolumic relaxation time ratio were independent predictors of stage C among echocardiographic indices in MMVD dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: Inappropriate VAC assessed by echocardiographically estimated Ea/Ees is associated with advanced disease severity in dogs with MMVD.
• Acute effects of intravenous pimobendan administration in dog models of chronic precapillary pulmonary hypertension

  T. Morita, K. Nakamura, T. Osuga, S. Kawamoto, S. Miki, K. Sasaoka, M. Takiguchi

  Journal of Veterinary Cardiology 32 16 - 27 1760-2734 2020/12 

  © 2020 Elsevier B.V. Introduction: Pulmonary hypertension (PH) is a progressive and potentially life-threatening disease. Several drugs are used for the treatment of dogs with precapillary PH. Pimobendan is an inotropic drug with phosphodiesterase 3 inhibitory and calcium-sensitizing effects. Pimobendan administration improved right ventricular (RV) function and lowered pulmonary arterial pressure in some human patients with precapillary PH. However, the efficacy of pimobendan in dogs with precapillary PH is unknown. Animals, material, and methods: An implantable port device was percutaneously placed in the cranial vena cava of five laboratory beagles. Chronic embolic precapillary PH was induced via the repeated injection of microspheres every 1–2 days. Microsphere injection was continued until systolic pulmonary arterial pressure reached 50 mmHg. Right heart catheterization and echocardiography were performed at baseline and after injections of placebo and pimobendan (0.15 mg/kg). Results: Repeated injections of microspheres caused an increase in pulmonary vascular resistance, a decrease in stroke volume, RV dilation, left ventricular (LV) and RV dysfunction, and RV dyssynchrony as assessed using echocardiography. Compared with placebo, pimobendan improved LV and RV function based on the LV Tei index from 0.48 to 0.38 (p=0.002) and the RV Tei index from 0.76 to 0.61 (p=0.008), as well as the stroke volume index from 29.4 to 36.7 ml/m2 (p=0.012), respectively. Conclusions: In dog models of chronic PH, intravenous pimobendan effectively improved RV and LV function and increased stroke volume. However, pimobendan administration did not decrease pulmonary arterial pressure or produce hypotension.
• Long interspersed nucleotide element-1 hypomethylation in canine malignant mucosal melanoma.

  Teita Ishizaki, Jumpei Yamazaki, Shinji Meagawa, Nozomu Yokoyama, Keisuke Aoshima, Mitsuyoshi Takiguchi, Takashi Kimura

  Veterinary and comparative oncology 18 (4) 854 - 860 2020/12 [Refereed]
   

  Canine malignant melanoma is a common cancer with a high mortality rate and is a clinically important disease. DNA methylation has been considered to be a potential tumorigenic mechanism through aberrant DNA methylation at promoter region which represses gene transcription. Global hypomethylation could also facilitate chromosome instability. There are few reports regarding DNA methylation in canine malignant melanoma; therefore, the purpose of this study was to examine DNA methylation status of long interspersed nucleotide element-1 (LINE-1) to be a surrogate marker of genome-wide methylation changes in this disease. We measured levels of DNA methylation of two adjacent cytosine-guanine sites on CpG island (CGI) at the putative promoter of canine LINE-1 sequence by bisulphite-pyrosequencing in 41 canine melanoma patient samples as well as six cell lines compared with normal mucosae. The survival rates were obtained from owners or medical records. We found DNA methylation levels of LINE-1 in normal mucosae were methylated. Interestingly, both melanoma cell lines and clinical melanoma samples showed remarkable hypomethylation. In addition, patients with lower LINE-1 methylation showed worse prognosis than those with higher LINE-1 methylation, though the difference did not reach statistical significance (P = .09). Here, we demonstrate that hypomethylation of LINE-1 is an epigenetically aberrant feature in canine melanoma with possible prognostic value.
• Wobbly hedgehog syndrome with disseminated histiocytic sarcoma and lateral ventricular meningioma in an African pygmy hedgehog.

  Lesa A Thompson, Atsuya Morita, Shoko Murakami, Noboru Sasaki, Miou Murashita, Ryou Yamazaki, Atsushi Kobayashi, Takashi Kimura, Mitsuyoshi Takiguchi

  Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 32 (6) 953 - 956 2020/11 [Refereed]
   

  An 8-mo-old male African pygmy hedgehog was anorectic and ataxic; physical examination revealed tetraparesis and a gangrenous left hindlimb. Analgesic and supportive care were administered, but the animal died 3 d after presentation. Postmortem examination revealed a histiocytic sarcoma in a mesenteric lymph node with metastasis to several organs, multifocal vacuolation in the cerebral and cerebellar white matter, and a meningioma in the left lateral ventricle. We diagnosed wobbly hedgehog syndrome (WHS) with disseminated histiocytic sarcoma and lateral ventricular meningioma. Ventricular meningioma, a rare neoplasm in veterinary and human patients, has not been reported previously in hedgehogs, to our knowledge. The neurologic signs in our case were probably caused by the WHS-related vacuolar lesions and are consistent with those of reported WHS cases. Duration of illness was shorter than is typical of WHS cases, which might be related to the disseminated histiocytic sarcoma. Clinical relevance of the lateral ventricular meningioma was not evident because the ventricular mass was localized and not invasive.
• Gene expression of leucine-rich alpha-2 glycoprotein in the polypoid lesion of inflammatory colorectal polyps in miniature dachshunds.

  Hiroshi Ohta, Yu Tamura, Nozomu Yokoyama, Noriyuki Nagata, Tatsuyuki Osuga, Noboru Sasaki, Yumiko Kagawa, Keitaro Morishita, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 82 (10) 1445 - 1449 2020/10/20 [Refereed]
   

  Inflammatory colorectal polyps (ICRPs) in miniature dachshunds (MDs) are speculated to be a breed-specific inflammatory bowel disease (IBD). Leucine-rich alpha-2 glycoprotein (LRG) has been identified as a novel biomarker of human IBD. The aim of this study was to examine LRG gene expression in the polypoid lesions of ICRPs. Polypoid lesion specimens were collected from 24 MDs with ICRPs. Nonpolypoid colonic mucosa was collected from 18 MDs with ICRPs and 10 controls. The gene expression of LRG, interleukin (IL)-1β, IL-6, tumor necrosis factor-α, and IL-22 was examined. The expression of LRG gene was significantly increased in the polypoid lesions of ICRPs and correlated with that of the four cytokines. In conclusion, the LRG gene was expressed within the polypoid lesions of ICRPs and might be associated with local cytokine expression.
• Altered hepatic cytochrome P450 expression in cats after chronic exposure to decabromodiphenyl ether (BDE-209).

  Kraisiri Khidkhan, Hazuki Mizukawa, Yoshinori Ikenaka, Shouta M M Nakayama, Kei Nomiyama, Nozomu Yokoyama, Osamu Ichii, Mitsuyoshi Takiguchi, Shinsuke Tanabe, Mayumi Ishizuka

  The Journal of veterinary medical science 82 (7) 978 - 982 2020/07/31 [Refereed]
   

  The knowledge of cytochrome P450 (CYP) expression involved in chemical exposure are necessary in clinical applications for the medication and prediction of adverse effects. The aim of this study was to evaluate the mRNA expression of CYP1-CYP3 families in cats exposed to BDE-209 for one year. All selected CYP isoforms showed no significant difference in mRNA expressions between control and exposure groups, however, CYP3A12 and CYP3A131 revealed tend to be two times higher in the exposure group compared to control group. The present results indicate that the chronic exposure of BDE209 could not alter CYP expression in the liver of cats. This result considered caused by the deficiency of CYP2B subfamily which is major metabolism enzyme of polybrominated diphenyl ethers (PBDEs) in cat.
• An area ratio of thyroid gland to common carotid artery for evaluating the thyroid gland size.

  Noboru Sasaki, Noriyuki Nagata, Keitaro Morishita, Tatsuyuki Osuga, Kazuyoshi Sasaoka, Nozomu Yokoyama, Hiroshi Ohta, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 82 (7) 1012 - 1016 2020/07/31 

  The feasibility of ultrasonographic measurement of thyroid gland area to common carotid artery (TG:CCA) was investigated. Twenty-one healthy, 12 hypothyroid and 18 non-thyroid illness (NTI) dogs were evaluated. The area of thyroid lobe and common carotid artery in right and left sides were measured using the same ultrasonographic images in transverse plane. The average of the right and left ratio was calculated as TG:CCA. The median TG:CCA of 21 healthy dogs was 1.53, and it did not correlate either body weight or age. The median TG:CCA of 12 hypothyroid dogs was 0.81, which was significantly lower than that of 18 NTI dogs (1.81, P<0.001). If the cut off value <1.12 was used, TG:CCA indicated hypothyroidism with a sensitivity of 100%, specificity of 83%, and accuracy of 90%. Our data indicated that TG:CCA was independent of both body weight, which may contribute to consistent measurement of thyroid size. The results of this study suggest that TG:CCA is a promising tool for diagnosing canine hypothyroidism.
• Activities of matrix metalloproteinase-2, matrix metalloproteinase-9, and serine proteases in samples of the colorectal mucosa of Miniature Dachshunds with inflammatory colorectal polyps.

  Noriyuki Nagata, Hiroshi Ohta, Arisa Yamada, Yong Bin Teoh, Osamu Ichii, Keitaro Morishita, Noboru Sasaki, Mitsuyoshi Takiguchi

  American journal of veterinary research 81 (7) 572 - 580 2020/07 [Refereed][Not invited]
   

  OBJECTIVE: To investigate the activities of gelatinases (matrix metalloproteinase [MMP]-2 and MMP-9) and serine proteases in the colorectal mucosa of Miniature Dachshunds (MDs) with inflammatory colorectal polyps (ICRPs). ANIMALS: 15 MDs with ICRPs and 5 dogs with non-ICRP-related large bowel diarrhea (controls). PROCEDURES: Zymographic methods were used to evaluate the activities of MMP-2, MMP-9, latent forms of MMP-2 and MMP-9 (pro-MMP-2 and pro-MMP-9), and serine proteases in inflamed and noninflamed tissue samples from MDs with ICRPs and in noninflamed tissue samples from control dogs. The associations of serine protease activities with MMP-2 or MMP-9 activity were also analyzed. RESULTS: Activities of pro-MMP-2 and pro-MMP-9 were detected in most tissue samples, regardless of the tissue type, whereas activities of MMP-2 and MMP-9 were not detected in control tissue samples. In the inflamed tissue samples from MDs with ICRPs, the activities of MMP-2, pro-MMP-9, and MMP-9 were significantly higher than those in the noninflamed tissue samples from those dogs. Serine protease activities were significantly higher in the inflamed and noninflamed tissue samples from MDs with ICRP, compared with findings for control tissue samples. A weak correlation was detected between serine protease activities and MMP-9 activity. CONCLUSIONS AND CLINICAL RELEVANCE: Study results suggested that gelatinase and serine protease activities are upregulated in the colorectal mucosa of MDs with ICRPs, possibly contributing to the pathogenesis of this disease through the functions of these enzymes in degradation of extracellular matrix and promotion of inflammatory cell migration and inflammatory responses.
• Wobbly hedgehog syndrome with disseminated histiocytic sarcoma and lateral ventricular meningioma in an African pygmy hedgehog

  Lesa A. Thompson, Atsuya Morita, Shoko Murakami, Noboru Sasaki, Miou Murashita, Ryou Yamazaki, Atsushi Kobayashi, Takashi Kimura, Mitsuyoshi Takiguchi

  J Vet Diagn Invest 2020/06 [Refereed][Not invited]
  
• Altered hepatic cytochrome P450 expression in cats after chronic exposure to decabromodiphenyl ether (BDE-209).

  Kraisiri Khidkhan, Hazuki Mizukawa, Yoshinori Ikenaka, Shouta M M Nakayama, Kei Nomiyama, Nozomu Yokoyama, Osamu Ichii, Mitsuyoshi Takiguchi, Shinsuke Tanabe, Mayumi Ishizuka

  The Journal of veterinary medical science 82 (7) 978 - 982 2020/05/21 [Refereed][Not invited]
   

  The knowledge of cytochrome P450 (CYP) expression involved in chemical exposure are necessary in clinical applications for the medication and prediction of adverse effects. The aim of this study was to evaluate the mRNA expression of CYP1-CYP3 families in cats exposed to BDE-209 for one year. All selected CYP isoforms showed no significant difference in mRNA expressions between control and exposure groups, however, CYP3A12 and CYP3A131 revealed tend to be two times higher in the exposure group compared to control group. The present results indicate that the chronic exposure of BDE209 could not alter CYP expression in the liver of cats. This result considered caused by the deficiency of CYP2B subfamily which is major metabolism enzyme of polybrominated diphenyl ethers (PBDEs) in cat.
• Genome-wide DNA methylation analysis in canine gastrointestinal lymphoma.

  Hiroshi Ohta, Jumpei Yamazaki, Jaroslav Jelinek, Teita Ishizaki, Yumiko Kagawa, Nozomu Yokoyama, Noriyuki Nagata, Noboru Sasaki, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 82 (5) 632 - 638 2020/03/25 [Refereed][Not invited]
   

  DNA methylation is the covalent modification of methyl groups to DNA mostly at CpG dinucleotides and one of the most studied epigenetic mechanisms that leads to gene expression variability without affecting the DNA sequence. Genome-wide analysis of DNA methylation identified the signatures that could define subtypes of human lymphoma patients. The objective of this study was to conduct the genome-wide analysis of DNA methylation in dogs with gastrointestinal lymphoma (GIL). Genomic DNA was extracted from endoscopic biopsies from 10 dogs with GIL. We performed Digital Restriction Enzyme Assay of DNA Methylation (DREAM) for genome-wide DNA methylation analysis that could provide highly quantitative information on DNA methylation levels of CpG sites across the dog genome. We successfully obtained data of quantitative DNA methylation level for 148,601-162,364 CpG sites per GIL sample. Next, we analyzed 83,132 CpG sites to dissect the differences in DNA methylation between GIL and normal peripheral blood mononuclear cells (PBMCs). We found 383-3,054 CpG sites that were hypermethylated in GIL cases compared to PBMCs. Interestingly, 773 CpG sites including promoter regions of 61 genes were identified to be commonly hypermethylated in more than half of the cases, suggesting conserved DNA methylation patterns that are abnormal in GIL. This study revealed that there was a large number of hypermethylated sites that are common in most of canine GIL. These abnormal DNA methylation could be involved in tumorigenesis of the canine GIL.
• Long interspersed nucleotide element-1 hypomethylation in canine malignant mucosal melanoma.

  Teita Ishizaki, Jumpei Yamazaki, Shinji Meagawa, Nozomu Yokoyama, Keisuke Aoshima, Mitsuyoshi Takiguchi, Takashi Kimura

  Veterinary and comparative oncology 18 (4) 854 - 860 2020/03/18 [Refereed][Not invited]
   

  Canine malignant melanoma is a common cancer with a high mortality rate and is a clinically important disease. DNA methylation has been considered to be a potential tumorigenic mechanism through aberrant DNA methylation at promoter region which represses gene transcription. Global hypomethylation could also facilitate chromosome instability. There are few reports regarding DNA methylation in canine malignant melanoma; therefore, the purpose of this study was to examine DNA methylation status of long interspersed nucleotide element-1 (LINE-1) to be a surrogate marker of genome-wide methylation changes in this disease. We measured levels of DNA methylation of two adjacent cytosine-guanine sites on CpG island (CGI) at the putative promoter of canine LINE-1 sequence by bisulphite-pyrosequencing in 41 canine melanoma patient samples as well as six cell lines compared with normal mucosae. The survival rates were obtained from owners or medical records. We found DNA methylation levels of LINE-1 in normal mucosae were methylated. Interestingly, both melanoma cell lines and clinical melanoma samples showed remarkable hypomethylation. In addition, patients with lower LINE-1 methylation showed worse prognosis than those with higher LINE-1 methylation, though the difference did not reach statistical significance (P = .09). Here, we demonstrate that hypomethylation of LINE-1 is an epigenetically aberrant feature in canine melanoma with possible prognostic value.
• Clinical characteristics of dogs with food-responsive protein-losing enteropathy.

  Noriyuki Nagata, Hiroshi Ohta, Nozomu Yokoyama, Yong Bin Teoh, Khoirun Nisa, Noboru Sasaki, Tatsuyuki Osuga, Keitaro Morishita, Mitsuyoshi Takiguchi

  Journal of veterinary internal medicine 34 (2) 659 - 668 2020/03 [Refereed][Not invited]
   

  BACKGROUND: In dogs with protein-losing enteropathy (PLE), data on the clinical characteristics of food-responsive PLE (FR-PLE) remain scarce. OBJECTIVE: To determine the clinical characteristics of FR-PLE in dogs responsive to ultralow-fat diet (ULFD) management. ANIMALS: Thirty-three dogs diagnosed with PLE based on standard diagnostic criteria. METHODS: Retrospective review of medical records. Clinical findings were compared between dogs with FR-PLE (FR-PLE group) and those with immunosuppressant-responsive PLE (IR-PLE) or nonresponsive PLE (NR-PLE) (IR/NR-PLE group). The area under the curve (AUC) of a receiver operating characteristic curve was used to evaluate the ability of factors to differentiate the FR-PLE and IR/NR-PLE groups. Survival time was compared between the FR-PLE and IR/NR-PLE groups. RESULTS: Twenty-three dogs responded to ULFD management and were diagnosed with FR-PLE. The canine chronic enteropathy clinical activity index (CCECAI) was significantly lower in the FR-PLE group than in the IR/NR-PLE group (P < .001). The AUC of CCECAI for differentiating the FR-PLE group was 0.935 (95% confidence interval [CI], 0.845-1.000) with an optimal cutoff value of 8 (sensitivity, 0.826; specificity, 0.889). Survival times were significantly longer in the FR-PLE group (median, not reached) than in the IR/NR-PLE group (median, 432 days; P < .001). CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs that respond to ULFD management and are diagnosed with FR-PLE are expected to have a favorable prognosis. Clinical scores, specifically the CCECAI, could be useful for differentiating FR-PLE from IR-PLE or NR-PLE.
• Computed tomographic features for differentiating benign from malignant liver lesions in dogs.

  Rommaneeya Leela-Arporn, Hiroshi Ohta, Genya Shimbo, Kiwamu Hanazono, Tatsuyuki Osuga, Keitaro Morishita, Noboru Sasaki, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 81 (12) 1697 - 1704 2019/12/18 [Refereed]
   

  Thus far, there are few computed tomography (CT) characteristics that can distinguish benign and malignant etiologies. The criteria are complex, subjective, and difficult to use in clinical applications due to the high level of experience needed. This study aimed to identify practical CT variables and their clinical relevance for broadly classifying histopathological diagnoses as benign or malignant. In this prospective study, all dogs with liver nodules or masses that underwent CT examination and subsequent histopathological diagnosis were included. Signalments, CT findings and histopathological diagnoses were recorded. Seventy liver nodules or masses in 57 dogs were diagnosed, comprising 18 benign and 52 malignant lesions. Twenty-three qualitative and quantitative CT variables were evaluated using univariate and stepwise multivariate analyses, respectively. Two variables, namely, the postcontrast enhancement pattern of the lesion in the delayed phase (heterogeneous; odds ratio (OR): 14.7, 95% confidence interval (CI): 0.82-262.03, P=0.0429) and the maximal transverse diameter of the lesion (>4.5 cm; OR: 33.3, 95% CI: 2.29-484.18, P=0.0006), were significantly related to the differentiation of benign from malignant liver lesions, with an area under the curve of 0.8910, representing an accuracy of 88.6%. These findings indicate that features from triple-phase CT can provide information for distinguishing pathological varieties of focal liver lesions and for clinical decision making. Evaluations of the maximal transverse diameter and postcontrast enhancement pattern of the lesion included simple CT features for predicting liver malignancy with high accuracy in clinical settings.
• Tissue distribution and characterization of feline cytochrome P450 genes related to polychlorinated biphenyl exposure.

  Khidkhan K, Mizukawa H, Ikenaka Y, Nakayama SMM, Nomiyama K, Yokoyama N, Ichii O, Darwish WS, Takiguchi M, Tanabe S, Ishizuka M

  Comparative biochemistry and physiology. Toxicology & pharmacology : CBP 226 108613 - 108613 1532-0456 2019/12 [Refereed][Not invited]
   

  Cats have been known to be extremely sensitive to chemical exposures. To understand these model species' sensitivity to chemicals and their toxicities, the expression profiles of xenobiotic-metabolizing enzymes should be studied. Unfortunately, the characterization of cytochrome P450 (CYP), the dominant enzyme in phase I metabolism, in cats has not extensively been studied. Polychlorinated biphenyls (PCBs) are known as CYP inducers in animals, but the information regarding the PCB-induced CYP expression in cats is limited. Therefore, in the present study, we aimed to elucidate the mRNA expression of the CYP1–CYP3 families in the cat tissues and to investigate the CYP mRNA expression related to PCB exposure. In cats, the greatest abundance of CYP1–CYP3 (CYP1A2, CYP2A13, CYP2C41, CYP2D6, CYP2E1, CYP2E2, CYP2F2, CYP2F5, CYP2J2, CYP2U1, and CYP3A132) was expressed in the liver, but some extrahepatic isozymes were found in the kidney (CYP1A1), heart (CYP1B1), lung (CYP2B11 and CYP2S1) and small intestine (CYP3A131). In cats, CYP1A1, CYP1A2 and CYP1B1 were significantly upregulated in the liver as well as in several tissues exposed to PCBs, indicating that these CYPs were distinctly induced by PCBs. The strong correlations between 3,3′,4,4′-tetrachlorobiphenyl (CB77) and CYP1A1 and CYP1B1 mRNA expressions were noted, demonstrating that CB77 could be a potent CYP1 inducer. In addition, these CYP isoforms could play an essential role in the PCBs biotransformation, particularly 3–4 Cl-PCBs, because a high hydroxylated metabolite level of 3–4 Cl-OH-PCBs was observed in the liver.
• Computed tomographic features for differentiating benign from malignant liver lesions in dogs.

  Leela-Arporn R, Ohta H, Shimbo G, Hanazono K, Osuga T, Morishita K, Sasaki N, Takiguchi M

  The Journal of veterinary medical science 81 (12) 1697 - 1704 0916-7250 2019/12 [Refereed][Not invited]
  
• Effects of PCB exposure on serum thyroid hormone levels in dogs and cats.

  Takaguchi K, Nishikawa H, Mizukawa H, Tanoue R, Yokoyama N, Ichii O, Takiguchi M, Nakayama SMM, Ikenaka Y, Kunisue T, Ishizuka M, Tanabe S, Iwata H, Nomiyama K

  The Science of the total environment 688 1172 - 1183 0048-9697 2019/10 [Refereed][Not invited]
   

  Polychlorinated biphenyls (PCBs) and their hydroxylated metabolites (OH-PCBs) might disrupt thyroid function. However, there is no clear evidence of PCB exposure disrupting thyroid hormone (TH) homeostasis in dogs and cats. The present study conducted in vivo experiments to evaluate the effects of a mixture of 12 PCB congeners (CB18, 28, 70, 77, 99, 101, 118, 138, 153, 180, 187 and 202, each congener 0.5 mg/kg BW, i.p. administration) on serum TH levels in male dogs (Canis lupus familiaris) and male cats (Felis silvestris catus). In PCB-exposed dogs, the time courses of higher-chlorinated PCBs and L-thyroxine (T4)-like OH-PCBs (4-OH-CB107 and 4-OH-CB202) concentrations were unchanged or tended to increase, whereas those of lower-chlorinated PCBs and OH-PCBs tended to decrease after 24 h. In PCB-exposed cats, concentrations of PCBs increased until 6 h and then remained unchanged. The levels of lower-chlorinated OH-PCBs including 4'-OH-CB18 increased until 96 h and then decreased. In PCB-exposed dogs, free T4 concentrations were higher than those in the control group at 48 and 96 h after PCB administration and positively correlated with the levels of T4-like OH-PCBs, suggesting competitive binding of T4 and T4-like OH-PCBs to a TH transporter, transthyretin. Serum levels of total T4 and total 3,3',5-triiodo-L-thyronine (T3) in PCB-exposed dogs were lower than in the control group at 24 and 48 h and negatively correlated with PCB concentrations, implying that PCB exposure enhanced TH excretion by increasing TH uptake and TH conjugation enzyme activities in the dog liver. In contrast, no obvious changes in TH levels were observed in PCB-exposed cats. This could be explained by the lower levels of T4-like OH-PCBs and lower hepatic conjugation enzyme activities in cats compared with dogs. Different effects on serum TH levels in PCB-exposed dogs and cats are likely to be attributable to species-specific PCB and TH metabolism.
• Change in right ventricular function in an American cocker spaniel with acute pulmonary thromboembolism.

  Tomoya Morita, Kensuke Nakamura, Tatsuyuki Osuga, Kiwamu Hanazono, Keitaro Morishita, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 81 (9) 1259 - 1265 0916-7250 2019/09/03 [Refereed][Not invited]
   

  A 12-year-old neutered female American cocker spaniel weighing 9.9 kg was presented for evaluation with a 2-day history of dyspnea and anorexia. Echocardiography revealed severe pulmonary hypertension (estimated systolic pulmonary arterial pressure, 93.4 mmHg) with right heart enlargement, pulmonary arterial dilation, and right ventricular dysfunction. The dilation of left heart and congenital cardiac shunt were not observed. Pulmonary thromboembolism (PTE) was confirmed by computed tomographic angiography. After treatment with antiplatelet and anticoagulant, the clinical sign and the echocardiographic abnormality of right heart were improved. These echocardiographic findings are not specific for PTE, but it can be useful as a rule-in test for PTE when other causes of pulmonary hypertension are excluded and a monitor of therapeutic efficacy.
• Usefulness of noninvasive shear wave elastography for the assessment of hepatic fibrosis in dogs with hepatic disease.

  Tamura M, Ohta H, Shimbo G, Osuga T, Sasaki N, Morishita K, Kagawa Y, Takiguchi M

  Journal of veterinary internal medicine 33 (5) 2067 - 2074 0891-6640 2019/09 [Refereed][Not invited]
  
• Effect of ultra low fat diet on regulatory T cells of dogs with intestinal limphangiectasia secondary to chronic enteropathy: A pilot study

  Khoirun Nisa, Hiroshi Ohta, Nozomu Yokoyama, Masahiro Tamura, Noriyuki Nagata, Kazuyoshi Sasaoka, Angkhana Dermlim, Rommaneeya Leela-Arporn, Tomoya Morita, Tatsuyuki Osuga, Noboru Sasaki, Keitaro Morishita, Kensuke Nakamura, Yumiko Kagawa, Mitsuyoshi Takiguchi

  JAPANESE JOURNAL OF VETERINARY RESEARCH 67 (3) 221 - 227 0047-1917 2019/08 [Refereed][Not invited]
   

  Ultra low fat diet (ULFD) has shown a good therapeutic effect in dogs with intestinal lymphangiectasia secondary to CE (IL-CE). As we hypothesized that the mechanism of ULFD may involve the resolution of immunological impairment in IL-CE dogs, our study aimed to investigate the effect of ULFD on the circulating regulatory T cell (Treg) of IL-CE dogs. Treg frequency of pre- and post-ULFD blood samples from dogs with IL-CE (n = 3) were measured using flow cytometry. Treg frequency in IL-CE group before ULFD treatment showed lower tendency when compared to healthy control (mean +/- SD; 2.0 +/- 1.6% and 4.3 +/- 1.4%, respectively), and showed increased tendency after ULFD (mean +/- SD; 4.5 +/- 3.5%). The results suggested that ULFD might restore circulating Treg frequency of IL-CE dogs.
• Findings of contrast-enhanced ultrasonography with Sonazoid for cholangiocellular adenoma in three dogs.

  Tamura M, Nakamura K, Osuga T, Shimbo G, Sasaki N, Morishita K, Ohta H, Takiguchi M

  The Journal of veterinary medical science 81 (8) 1104 - 1108 0916-7250 2019/08 [Refereed][Not invited]
  
• Plasma amino acid profiles in dogs with inflammatory bowel disease.

  Yu Tamura, Hiroshi Ohta, Yumiko Kagawa, Tatsuyuki Osuga, Keitaro Morishita, Noboru Sasaki, Mitsuyoshi Takiguchi

  Journal of veterinary internal medicine 33 (4) 1602 - 1607 2019/07 

  BACKGROUND: Lymphocytic-plasmacytic enteritis is the common form of idiopathic inflammatory bowel disease (IBD) in dogs. In human IBD, disturbances of amino acid metabolism have been demonstrated to be involved in the pathophysiology of IBD. Therefore, plasma amino acid profile might represent a novel marker of human IBD. OBJECTIVES: To determine the plasma amino acid profiles of dogs with IBD and its usefulness as a novel marker of IBD in dogs. ANIMALS: Fasting blood plasma was obtained from 10 dogs with IBD and 12 healthy dogs. METHODS: All IBD dogs were prospectively included in this study, and heparinized blood samples were collected. The plasma concentrations of 21 amino acids were determined using the ninhydrin method. The relationships among the plasma amino acid concentrations and plasma C-reactive protein (CRP) concentration, canine chronic enteropathy clinical activity index (CCECAI), and overall World Small Animal Veterinary Association (WSAVA) score were investigated. RESULTS: Median concentration (nmol/mL) of methionine [46.2; range, 30.0-59.3], proline [119.4; range, 76.7-189.2], serine [115.1; range, 61.4-155.9], and tryptophan [17.4; range, 11.9-56.3]) were significantly lower than in control dogs [62.6; range, 51.0-83.6, 199.1; range, 132.5-376.7, 164.3; range, 124.7-222.9, and 68.3; range, 35.7-94.8, respectively]. A negative correlation was identified between the plasma serine concentration and CCECAI (r s = -.67, P = .03), but there were no correlations between plasma amino acid concentrations and CRP concentration or overall WSAVA score. CONCLUSIONS AND CLINICAL IMPORTANCE: Plasma serine concentration might represent a novel maker of IBD in dogs.
• Plasma-free amino acid profiles in dogs with hepatocellular carcinoma.

  Leela-Arporn R, Ohta H, Tamura M, Nagata N, Sasaoka K, Dermlim A, Nisa K, Osuga T, Morishita K, Sasaki N, Takiguchi M

  Journal of veterinary internal medicine 33 (4) 1653 - 1659 0891-6640 2019/07 [Refereed][Not invited]
  
• Effect of acute volume loading on left atrial strain values derived from two-dimensional speckle tracking echocardiography in dogs.

  Dermlim A, Osuga T, Nakamura K, Morita T, Nisa K, Sasaoka K, Leela-Arporn R, Nagata N, Tamura M, Sasaki N, Ohta H, Takiguchi M

  The Journal of veterinary medical science 81 (7) 949 - 957 0916-7250 2019/07 [Refereed][Not invited]
  
• Epidemiology of massive hepatocellular carcinoma in dogs: A 4-year retrospective study.

  Leela-Arporn R, Ohta H, Nagata N, Sasaoka K, Tamura M, Dermlim A, Nisa K, Morishita K, Sasaki N, Nakamura K, Takagi S, Hosoya K, Takiguchi M

  Veterinary journal (London, England : 1997) 248 74 - 78 1090-0233 2019/06 [Refereed][Not invited]
  
• Right ventricular function and dyssynchrony measured by echocardiography in dogs with precapillary pulmonary hypertension

  T. Morita, K. Nakamura, T. Osuga, K. Morishita, N. Sasaki, H. Ohta, M. Takiguchi

  Journal of Veterinary Cardiology Elsevier {BV} 23 1 - 14 1760-2734 2019/06 [Refereed][Not invited]
  
• The repeatability and left atrial strain analysis obtained via speckle tracking echocardiography in healthy dogs.

  Dermlim, A, Nakamura, K, Morita, T, Osuga, T, Nisa, K, Sasaoka, K, Leela-arporn, R, Sasaki, N, Ohta, H, Takiguchi, M

  Journal of Veterinary Cardiology 23 (1) 69 - 80 1760-2734 2019/06 [Refereed][Not invited]
  
• Predictive factors of malignancy in dogs with focal liver lesions using clinical data and ultrasonographic features.

  Leela-Arporn R, Ohta H, Tamura M, Nagata N, Sasaoka K, Dermlim A, Nisa K, Morishita K, Sasaki N, Nakamura K, Takagi S, Hosoya K, Takiguchi M

  The Journal of veterinary medical science 81 (5) 723 - 729 0916-7250 2019/05 [Refereed][Not invited]
  
• Plasma amino acid profiles in dogs with inflammatory bowel disease.

  Tamura Y, Ohta H, Kagawa Y, Osuga T, Morishita K, Sasaki N, Takiguchi M

  Journal of veterinary internal medicine 0891-6640 2019/05 [Refereed][Not invited]
  
• Evaluation of liver and spleen stiffness of healthy dogs by use of two-dimensional shear wave elastography.

  Tamura M, Ohta H, Nisa K, Osuga T, Sasaki N, Morishita K, Takiguchi M

  American journal of veterinary research 80 (4) 378 - 384 0002-9645 2019/04 [Refereed][Not invited]
  
• Evaluation of duodenal perfusion by contrast-enhanced ultrasonography in dogs with chronic inflammatory enteropathy and intestinal lymphoma.

  Nisa K, Lim SY, Shinohara M, Osuga T, Yokoyama N, Tamura M, Nagata N, Sasaoka K, Dermlim A, Leela-Arporn R, Morita T, Sasaki N, Morishita K, Nakamura K, Ohta H, Takiguchi M

  Journal of Veterinary Internal Medicine 33 (2) 559 - 568 1939-1676 2019/03 [Refereed][Not invited]
   

  Background: Contrast-enhanced ultrasonography (CEUS) can be used to evaluate intestinal perfusion in healthy dogs. It is helpful for diagnosing and monitoring inflammatory bowel disease in humans and could be useful for dogs with chronic intestinal diseases. Objectives: To examine duodenal perfusion in dogs with chronic inflammatory enteropathy (CIE) and intestinal lymphoma. Animals: Client-owned dogs with CIE (n = 26) or intestinal lymphoma (n = 7) and dogs with gastrointestinal signs but histopathologically normal duodenum (controls, n = 14). Methods: In this cross-sectional study, dogs with CIE were classified into remission (n = 16) and symptomatic (n = 10) groups based on clinical scores determined at the time of CEUS. The duodenum was scanned after IV injection of Sonazoid® (0.01 mL/kg). CEUS-derived perfusion parameters, including time-to-peak, peak intensity (PI), area under the curve (AUC), and wash-in and wash-out rates were evaluated. Results: The PI was significantly higher in the symptomatic CIE group (median (range); 105.4 (89.3-128.8) MPV) than in the control group (89.9 (68.5-112.2) MPV). The AUC was significantly higher in the symptomatic CIE group (4847.9 (3824.3-8462.8) MPV.sec) than in the control (3448.9 (1559.5-4736.9) MPV.sec) and remission CIE (3862.3 (2094.5-6899.0) MPV.sec) groups. The PI and clinical score were positively correlated in the CIE group. No significant differences in perfusion parameters were detected between the lymphoma and CIE groups or the lymphoma and control groups. Conclusions and Clinical Importance: The PI and AUC can detect duodenal inflammation and hence are potentially useful for excluding a diagnosis of CIE.
• Contrast-enhanced ultrasonography is a feasible technique for quantifying hepatic microvascular perfusion in dogs with extrahepatic congenital portosystemic shunts.

  Tamura M, Ohta H, Nisa K, Osuga T, Sasaki N, Morishita K, Takiguchi M

  Veterinary radiology & ultrasound : the official journal of the American College of Veterinary Radiology and the International Veterinary Radiology Association 60 (2) 192 - 200 1058-8183 2019/03 [Refereed][Not invited]
  
• Evaluation of duodenal perfusion by contrast-enhanced ultrasonography in dogs with chronic inflammatory enteropathy and intestinal lymphoma.

  Nisa K, Lim SY, Shinohara M, Osuga T, Yokoyama N, Tamura M, Nagata N, Sasaoka K, Dermlim A, Leela-Arporn R, Morita T, Sasaki N, Morishita K, Nakamura K, Ohta H, Takiguchi M

  Journal of veterinary internal medicine 33 (2) 559 - 568 0891-6640 2019/03 [Refereed][Not invited]
   

  Background: Contrast-enhanced ultrasonography (CEUS) can be used to evaluate intestinal perfusion in healthy dogs. It is helpful for diagnosing and monitoring inflammatory bowel disease in humans and could be useful for dogs with chronic intestinal diseases. Objectives: To examine duodenal perfusion in dogs with chronic inflammatory enteropathy (CIE) and intestinal lymphoma. Animals: Client-owned dogs with CIE (n = 26) or intestinal lymphoma (n = 7) and dogs with gastrointestinal signs but histopathologically normal duodenum (controls, n = 14). Methods: In this cross-sectional study, dogs with CIE were classified into remission (n = 16) and symptomatic (n = 10) groups based on clinical scores determined at the time of CEUS. The duodenum was scanned after IV injection of Sonazoid® (0.01 mL/kg). CEUS-derived perfusion parameters, including time-to-peak, peak intensity (PI), area under the curve (AUC), and wash-in and wash-out rates were evaluated. Results: The PI was significantly higher in the symptomatic CIE group (median (range); 105.4 (89.3-128.8) MPV) than in the control group (89.9 (68.5-112.2) MPV). The AUC was significantly higher in the symptomatic CIE group (4847.9 (3824.3-8462.8) MPV.sec) than in the control (3448.9 (1559.5-4736.9) MPV.sec) and remission CIE (3862.3 (2094.5-6899.0) MPV.sec) groups. The PI and clinical score were positively correlated in the CIE group. No significant differences in perfusion parameters were detected between the lymphoma and CIE groups or the lymphoma and control groups. Conclusions and Clinical Importance: The PI and AUC can detect duodenal inflammation and hence are potentially useful for excluding a diagnosis of CIE.
• Low-intensity pulsed ultrasound modifies effects of a myelin-related growth inhibitor and enhances neurite re-growth

  Noboru Sasaki, Nobuki Kudo, Hiroshi Ohta, Mitsuyoshi Takiguchi

  2019 IEEE INTERNATIONAL ULTRASONICS SYMPOSIUM (IUS) 1308 - 1310 1948-5719 2019 

  Axon regeneration is crucial for the functional recovery after spinal cord injury. Myelin-related growth inhibitors, such as Nogo-A, inhibit neurites regrowth and elongation. Low-intensity pulsed ultrasound (LIPUS) is known to show neuroprotective effects in neurodegenerative diseases. In the present study, we evaluated the feasibility of LIPUS for enhancing axon regeneration in the presence of Nogo-A. Low-intensity pulsed ultrasound was exposed to rat cortical neurons in vitro after Nogo-A treatment. The total length of neurites per cell in LIPUS group was significantly larger than that of non-LIPUS group. There was no significant difference in the number of neurites. The results suggest that LIPUS may enhance the elongation of neurites in the presence of Nogo-A while may not affect on neurites sprouting and branching. This preliminary study implies the feasibility of LIPUS for axonal regeneration.
• Corrigendum: Urinary Exosome-Derived microRNAs Reflecting the Changes in Renal Function in Cats.

  Ichii O, Ohta H, Horino T, Nakamura T, Hosotani M, Mizoguchi T, Morishita K, Nakamura K, Sasaki N, Takiguchi M, Sato R, Oyamada K, Elewa YHA, Kon Y

  Frontiers in veterinary science 6 2  2019 [Refereed][Not invited]
  
• Effect of acute volume overload on echocardiographic indices of right ventricular function and dyssynchrony assessed by use of speckle tracking echocardiography in healthy dogs.

  Morita T, Nakamura K, Osuga T, Yokoyama N, Morishita K, Sasaki N, Ohta H, Takiguchi M

  American journal of veterinary research 80 (1) 51 - 60 0002-9645 2019/01 [Refereed][Not invited]
   

  OBJECTIVE To determine the relationship between acute volume overload and echocardiographic indices of right ventricular (RV) function and dyssynchrony in healthy dogs. ANIMALS 7 healthy Beagles. PROCEDURES Right heart catheterization and echocardiography were performed in 7 healthy anesthetized Beagles at baseline and after induction of volume overload. Volume overload was induced by IV infusion of lactated Ringer solution (150 mL/kg/h for 90 minutes). Echocardiographic indices of RV function, including peak velocity of systolic tricuspid annular motion, tricuspid annulus plane systolic excursion, fractional area change, RV Tei index, RV longitudinal strain (RVLS), and systolic RV longitudinal strain rate (RVLSR), were obtained by use of speckle tracking echocardiography (STE). In addition, SD of the systolic shortening time of the right ventricle for the 6 segments (RV-SD6) was determined with STE. RESULTS Volume overload significantly increased the RV end-diastolic pressure, compared with the baseline value. Echocardiographic indices of RV function, except for septal RVLSR, were significantly enhanced by volume overload. In contrast, RV-SD6 did not change with volume overload. Although echocardiographic indices of RV function, except for septal RVLSR, were correlated with RV end-diastolic pressure, RV-SD6 was not correlated. CONCLUSIONS AND CLINICAL RELEVANCE Echocardiographic indices of RV function, including RVLS and RVLSR, were affected by acute short-term volume overload. Therefore, results for assessment of RV function by use of STE in dogs with clinical conditions associated with right-sided chronic volume overload, such as tricuspid and pulmonic regurgitation, should be interpreted with caution.
• Effects of immunosuppressive prednisolone therapy on pancreatic tissue and concentration of canine pancreatic lipase immunoreactivity in healthy dogs.

  Ohta H, Kojima K, Yokoyama N, Sasaki N, Kagawa Y, Hanazono K, Ishizuka T, Morishita K, Nakamura K, Takagi S, Takiguchi M

  Canadian journal of veterinary research = Revue canadienne de recherche veterinaire 82 (4) 278 - 286 0830-9000 2018/10 [Refereed][Not invited]
   

  The objective of this study was to examine the effects of immunosuppressive prednisolone therapy on pancreatic tissue and the concentration of serum canine pancreatic lipase immunoreactivity (cPLI) in healthy dogs. Six healthy beagle dogs were subcutaneously administered an immunosuppressive dose of prednisolone [4 mg/kg body weight (BW)] once daily for either 2 or 3 weeks. Serum cPLI concentration was measured before and after treatment. Ultrasonographic examination of the pancreas and laparoscopic biopsy and histopathological examination of the right pancreatic lobe and the liver were also conducted before and after treatment. The expression of pancreatic lipase messenger ribonucleic acid (mRNA) in the pancreas and liver was examined by polymerase chain reaction (PCR). Although the serum cPLI concentration was significantly higher on day 14 and on the day of the second laparoscopy than before treatment, it was classified as normal (<= 200 mu g/L) in 5 dogs and as abnormal (>= 400 mu g/L) in only 1 dog. None of the 6 dogs showed clinical signs of pancreatitis during the study period. After treatment, ultrasonographic examination of the pancreas showed no changes except for a hypoechoic pancreas in 1 dog. Histopathological examination of the right pancreatic lobe in all dogs showed no evidence of pancreatitis after treatment. Pancreatic lipase mRNA expression was detected in the pancreas, but not in the liver, before and after treatment. The administration of 4 mg/kg BW per day of prednisolone for 2 or 3 weeks increased the serum cPLI concentration without clinical signs of pancreatitis, although an abnormal cPLI concentration (>= 400 mu g/L) was observed in only 1 dog. No ultrasonographic or histological evidence of pancreatitis was observed in any of the dogs.
• ECG of the Month.

  Osuga T, Nakamura K, Morita T, Watanabe K, Takiguchi M

  Journal of the American Veterinary Medical Association 253 (4) 417 - 420 0003-1488 2018/08 [Refereed][Not invited]
  
• Pulmonary hypertension due to unclassified interstitial lung disease in a Pembroke Welsh corgi.

  Morita T, Nakamura K, Osuga T, Kobayashi A, Ichii O, Yabuki A, Takiguchi M

  The Journal of veterinary medical science 80 (6) 939 - 944 0916-7250 2018/06 [Refereed][Not invited]
  
• The effect of sedation with a combination of butorphanol and midazolam on quantitative contrast-enhanced ultrasonography of duodenum in healthy dogs.

  Nisa K, Lim SY, Osuga T, Yokoyama N, Tamura M, Nagata N, Sasaoka K, Dermlim A, Leela-Arporn R, Morita T, Sasaki N, Morishita K, Nakamura K, Ohta H, Takiguchi M

  The Journal of Veterinary Medical Science 80 (3) 453 - 459 0021-5295 2018/03 [Refereed][Not invited]
   

  Quantitative contrast-enhanced ultrasonography (CEUS) enables non-invasive and objective evaluation of intestinal perfusion by quantifying the intensity of enhancement on the intestine after microbubble contrast administration. During CEUS scanning, sedation is sometimes necessary to maintain animal cooperation. Nevertheless, the effect of sedative administration on the canine intestinal CEUS is unknown. This study aimed to investigate the effect of sedation with a combination of butorphanol and midazolam on the duodenal CEUS-derived perfusion parameters of healthy dogs. For this purpose, duodenum was imaged following contrast administration (Sonazoid®, 0.01 ml/kg) in six healthy beagles before and after intravenous injection of a combination of butorphanol (0.2 mg/kg) and midazolam (0.1 mg/kg). Furthermore, hemodynamic parameters including blood pressure and heart rate were recorded during the procedure. Five CEUS derived perfusion parameters including time-to-peak (TTP), peak intensity (PI), area under the curve (AUC), wash-in and wash-out rates (WiR and WoR, respectively) before and after sedation were statistically compared. The result showed that no significant change was detected in any of perfusion parameters. Systolic and mean arterial pressures significantly reduced after sedative administration, but diastolic arterial pressure and heart rate did not significantly change. Moreover, no significant partial correlation was observed between perfusion parameters and hemodynamic parameters. Thus, we concluded that the combination did not cause significant influence in duodenal CEUS perfusion parameters and could be a good option for sedation prior to duodenal CEUS in debilitated dogs.
• The effect of sedation with a combination of butorphanol and midazolam on quantitative contrast-enhanced ultrasonography of duodenum in healthy dogs

  Nisa K, Lim S.Y, Osuga T, Yokoyama N, Tamura M, Nagata N, Sasaoka K, Dermlim A, Leela-Arporn R, Morita T, Sasaki N, Morishita K, Nakamura K, Ohta H, Takiguchi M

  Journal of Veterinary Medical Science 80 (3) 453 - 459 0916-7250 2018/03 [Refereed][Not invited]
   

  Quantitative contrast-enhanced ultrasonography (CEUS) enables non-invasive and objective evaluation of intestinal perfusion by quantifying the intensity of enhancement on the intestine after microbubble contrast administration. During CEUS scanning, sedation is sometimes necessary to maintain animal cooperation. Nevertheless, the effect of sedative administration on the canine intestinal CEUS is unknown. This study aimed to investigate the effect of sedation with a combination of butorphanol and midazolam on the duodenal CEUS-derived perfusion parameters of healthy dogs. For this purpose, duodenum was imaged following contrast administration (Sonazoid®, 0.01 ml/kg) in six healthy beagles before and after intravenous injection of a combination of butorphanol (0.2 mg/kg) and midazolam (0.1 mg/kg). Furthermore, hemodynamic parameters including blood pressure and heart rate were recorded during the procedure. Five CEUS derived perfusion parameters including time-to-peak (TTP), peak intensity (PI), area under the curve (AUC), wash-in and wash-out rates (WiR and WoR, respectively) before and after sedation were statistically compared. The result showed that no significant change was detected in any of perfusion parameters. Systolic and mean arterial pressures significantly reduced after sedative administration, but diastolic arterial pressure and heart rate did not significantly change. Moreover, no significant partial correlation was observed between perfusion parameters and hemodynamic parameters. Thus, we concluded that the combination did not cause significant influence in duodenal CEUS perfusion parameters and could be a good option for sedation prior to duodenal CEUS in debilitated dogs.
• Urinary Exosome-Derived microRNAs Reflecting the Changes in Renal Function in Cats.

  Ichii O, Ohta H, Horino T, Nakamura T, Hosotani M, Mizoguchi T, Morishita K, Nakamura K, Sasaki N, Takiguchi M, Sato R, Oyamada K, Elewa YHA, Kon Y

  Frontiers in veterinary science 5 289  2018 [Refereed][Not invited]
  
• Transcranial Doppler Ultrasound Examination in Dogs with Suspected Intracranial Hypertension Caused by Neurologic Diseases

  Sasaoka K, Nakamura K, Osuga T, Morita T, Yokoyama N, Morishita K, Sasaki N, Ohta H, Takiguchi M

  Journal of Veterinary Internal Medicine 32 (1) 314 - 323 0891-6640 2018 [Refereed][Not invited]
   

  Background: Transcranial Doppler ultrasound examination (TCD) is a rapid, noninvasive technique used to evaluate cerebral blood flow and is useful for the detection of intracranial hypertension in humans. However, the clinical usefulness of TCD in diagnosing intracranial hypertension has not been demonstrated for intracranial diseases in dogs. Objectives: To determine the association between the TCD variables and intracranial hypertension in dogs with intracranial diseases. Animals: Fifty client-owned dogs with neurologic signs. Methods: Cross-sectional study. All dogs underwent TCD of the basilar artery under isoflurane anesthesia after magnetic resonance imaging (MRI). Dogs were classified into 3 groups based on MRI findings: no structural diseases (group I), structural disease without MRI evidence of intracranial hypertension (group II), and structural disease with MRI evidence of intracranial hypertension (group III). The TCD vascular resistance variables (resistive index [RI], pulsatility index [PI], and the ratio of systolic to diastolic mean velocity [Sm/Dm]) were measured. Results: Fifteen, 22, and 13 dogs were classified into groups I, II, and III, respectively. Dogs in group III had significantly higher Sm/Dm (median, 1.78; range, 1.44–2.58) than those in group I (median, 1.63; range, 1.43–1.75) and group II (median, 1.62; range, 1.27–2.10). No significant differences in RI and PI were identified among groups. Conclusions and Clinical Importance: Our findings suggest that increased Sm/Dm is associated with MRI findings of suspected intracranial hypertension in dogs with intracranial diseases and that TCD could be a useful tool to help to diagnose intracranial hypertension.
• Intracellular diminazene aceturate content and adenosine incorporation in diminazene aceturate-resistant Babesia gibsoni isolate in vitro

  Masahiro Yamasaki, Nao Watanabe, Natsuki Idaka, Tohru Yamamori, Ken-ichi Otsuguro, Naohiro Uchida, Aiko Iguchi, Hiroshi Ohta, Mitsuyoshi Takiguchi

  EXPERIMENTAL PARASITOLOGY 183 92 - 98 0014-4894 2017/12 [Refereed][Not invited]
   

  The mechanism of the development of diminazene aceturate (DA) resistance in Babesia gibsoni is still unknown even though DA-resistant B. gibsoni isolate was previously developed in vitro. To clarify the mechanisms of DA-resistance in B. gibsoni, we initially examined the intracellular DA content in the DA resistant isolate using high-performance liquid chromatography, and compared it with that in the wild type. As a result, the intracellular DA content in the DA-resistant isolate was significantly lower than that in the wild-type, suggesting that the decreased DA content may contribute to DA-resistance. Additionally, the glucose consumption of the DA-resistant isolate was significantly higher than that of the wild-type, indicating that a large amount of glucose is utilized to maintain DA-resistance. It is possible that a large amount of energy is utilized to maintain the mechanisms of DA-resistance. It was reported that as the structure of DA is similar with that of adenosine, DA may be taken up by the P2 transporter, which contributes to the uptake of adenosine, in Trypanosoma brucei brucei, and that the uptake of adenosine is decreased in DA-resistant T brucei brucei. In the present study, the adenosine incorporation in the DA-resistant B. gibsoni isolate was higher than in the wild-type. Moreover, the adenosine incorporation in the wild-type was not inhibited by the presence of DA. These results suggest that adenosine transport in B. gibsoni is not affected by DA and may not mediate DA-resistance. To clarify the mechanism of the development of DA resistance in B. gibsoni, we should investigate the cause of the decreased DA content in the DA-resistant isolate in the future. (C) 2017 Elsevier Inc. All rights reserved.
• Case report: Congenital methemoglobinemia in a cat with the reduced NADH-cytochrome b5 reductase 3 activity and missense mutations in CYB5R3

  Akiyoshi Tani, Jumpei Yamazaki, Kensuke Nakamura, Mitsuyoshi Takiguchi, Mutsumi Inaba

  Japanese Journal of Veterinary Research Faculty of Veterinary Medicine, Hokkaido University 65 (4) 201 - 206 0047-1917 2017/11 [Refereed][Not invited]
   

  © 2017, Hokkaido University. All rights reserved. A one-year-old castrated male mixed-breed cat was referred for detailed examination of long-term pale mucous membrane without any clinical episodes that may cause cyanosis. While no causative abnormalities were detected in thoracic radiography and echocardiography, and arterial partial pressure of oxygen was within normal value, methemoglobin concentration of the cat was increased to 30% and cytochrome b5 reductase activity, which converts methemoglobin to hemoglobin, was reduced to 2.0 IU/gHb (13.4 ± 1.7 IU/gHb in control cats, n = 3), indicating congenital methemoglobinemia. Sequence analysis of CYB5R3, which codes cytochrome b5 reductase, showed two missense mutations found in the patient, one of which was predicted to affect protein function.
• Spatial and temporal profile of cisplatin delivery by ultrasound-assisted intravesical chemotherapy in a bladder cancer model

  Noboru Sasaki, Kazuhiro Ishi, Nobuki Kudo, Shouta M. M. Nakayama, Kensuke Nakamura, Keitaro Morishita, Hiroshi Ohta, Mayumi Ishizuka, Mitsuyoshi Takiguchi

  PLOS ONE 12 (11) e0188093  1932-6203 2017/11 [Refereed][Not invited]
   

  Non-muscle invasive bladder cancer is one of the most common tumors of the urinary tract. Despite the current multimodal therapy, recurrence and progression of disease have been challenging problems. We hereby introduced a new approach, ultrasound-assisted intravesical chemotherapy, intravesical instillation of chemotherapeutic agents and microbubbles followed by ultrasound exposure. We investigated the feasibility of the treatment for nonmuscle invasive bladder cancer. In order to evaluate intracellular delivery and cytotoxic effect as a function to the thickness, we performed all experiments using a bladder cancer mimicking 3D culture model. Ultrasound-triggered microbubble cavitation increased both the intracellular platinum concentration and the cytotoxic effect of cisplatin at the thickness of 70 and 122 mu m of the culture model. The duration of enhanced cytotoxic effect of cisplatin by ultrasound-triggered microbubble cavitation was approximately 1 hr. Based on the distance and duration of delivery, we further tested the feasibility of repetition of the treatment. Triple treatment increased the effective distance by 1.6-fold. Our results clearly showed spatial and temporal profile of delivery by ultrasound-triggered microbubble cavitation in a tumor-mimicking structure. Furthermore, we demonstrated that the increase in intracellular concentration results in the enhancement of the cytotoxic effect in a structure with the certain thickness. Repetition of ultrasound exposure would be treatment of choice in future clinical application. Our results suggest ultrasound-triggered microbubble cavitation can be repeatable and is promising for the local control of non-muscle invasive bladder cancer.
• Diastolic heart failure associated with hemangiosarcoma infiltrating left ventricular walls in a dog

  Osuga T, Nakamura K, Morita T, Kagawa Y, Ohta H, Takiguchi M

  Canadian Veterinary Journal 58 (11) 1167 - 1170 0008-5286 2017/11 [Refereed][Not invited]
  
• Repeatability and reproducibility of quantitative contrast-enhanced ultrasonography for assessing duodenal perfusion in healthy dogs.

  Nisa K, Lim SY, Shinohara M, Nagata N, Sasaoka K, Dermlim A, Leela-Arporn R, Morita T, Yokoyama N, Osuga T, Sasaki N, Morishita K, Nakamura K, Ohta H, Takiguchi M

  The Journal of Veterinary Medical Science 79 (9) 1585 - 1590 0021-5295 2017/09 [Refereed][Not invited]
   

  Contrast-enhanced ultrasonography (CEUS) with microbubbles as a contrast agent allows the visualization and quantification of tissue perfusion. The assessment of canine intestinal perfusion by quantitative CEUS may provide valuable information for diagnosing and monitoring chronic intestinal disorders. This study aimed to assess the repeatability (intraday variability) and reproducibility (interday variability) of quantitative duodenal CEUS in healthy dogs. Six healthy beagles underwent CEUS three times within one day (4-hr intervals) and on two different days (1-week interval). All dogs were sedated with a combination of butorphanol (0.2 mg/kg) and midazolam (0.1 mg/kg) prior to CEUS. The contrast agent (Sonazoid®) was administered using the intravenous bolus method (0.01 ml/kg) for imaging of the duodenum. Time-intensity curves (TIC) were created by drawing multiple regions of interest (ROIs) in the duodenal mucosa, and perfusion parameters, including the time-to-peak (TTP), peak intensity (PI), area under the curve (AUC), and wash-in and wash-out rates (WiR and WoR, respectively), were generated. Intraday and interday coefficients of variation (CVs) for TTP, PI, AUC, WiR and WoR were <25% (range, 2.27-23.41%), which indicated that CEUS was feasible for assessing duodenal perfusion in healthy sedated dogs. A further study of CEUS in dogs with chronic intestinal disorders is necessary to evaluate its clinical applicability.
• Repeatability and reproducibility of quantitative contrast-enhanced ultrasonography for assessing duodenal perfusion in healthy dogs

  Nisa K, Lim S.Y, Shinohara M, Nagata N, Sasaoka K, Dermlim A, Leela-Arporn R, Morita T, Yokoyama N, Osuga T, Sasaki N, Morishita K, Nakamura K, Ohta H, Takiguchi M

  Journal of Veterinary Medical Science 79 (9) 1585 - 1590 0916-7250 2017/09 [Refereed][Not invited]
   

  Contrast-enhanced ultrasonography (CEUS) with microbubbles as a contrast agent allows the visualization and quantification of tissue perfusion. The assessment of canine intestinal perfusion by quantitative CEUS may provide valuable information for diagnosing and monitoring chronic intestinal disorders. This study aimed to assess the repeatability (intraday variability) and reproducibility (interday variability) of quantitative duodenal CEUS in healthy dogs. Six healthy beagles underwent CEUS three times within one day (4-hr intervals) and on two different days (1-week interval). All dogs were sedated with a combination of butorphanol (0.2 mg/kg) and midazolam (0.1 mg/kg) prior to CEUS. The contrast agent (Sonazoid®) was administered using the intravenous bolus method (0.01 ml/kg) for imaging of the duodenum. Time-intensity curves (TIC) were created by drawing multiple regions of interest (ROIs) in the duodenal mucosa, and perfusion parameters, including the time-to-peak (TTP), peak intensity (PI), area under the curve (AUC), and wash-in and wash-out rates (WiR and WoR, respectively), were generated. Intraday and interday coefficients of variation (CVs) for TTP, PI, AUC, WiR and WoR were <25% (range, 2.27-23.41%), which indicated that CEUS was feasible for assessing duodenal perfusion in healthy sedated dogs. A further study of CEUS in dogs with chronic intestinal disorders is necessary to evaluate its clinical applicability.
• Brain micro-inflammation at specific vessels dysregulates organ-homeostasis via the activation of a new neural circuit

  Yasunobu Arima, Takuto Ohki, Naoki Nishikawa, Kotaro Higuchi, Mitsutoshi Ota, Yuki Tanaka, Junko Nio-Kobayashi, Mohamed Elfeky, Ryota Sakai, Yuki Mori, Tadafumi Kawamoto, Andrea Stofkova, Yukihiro Sakashita, Yuji Morimoto, Masaki Kuwatani, Toshihihiko Iwanaga, Yoshichika Yoshioka, Naoya Sakamoto, Akihiko Yoshimura, Mitsuyoshi Takiguchi, Saburo Sakoda, Marco Prinz, Daisuke Kamimura, Masaaki Murakami

  ELIFE 6 2050-084X 2017/08 [Refereed][Not invited]
   

  Impact of stress on diseases including gastrointestinal failure is well-known, but molecular mechanism is not understood. Here we show underlying molecular mechanism using EAE mice. Under stress conditions, EAE caused severe gastrointestinal failure with high-mortality. Mechanistically, autoreactive-pathogenic CD4+ T cells accumulated at specific vessels of boundary area of third-ventricle, thalamus, and dentate-gyrus to establish brain micro-inflammation via stress gateway reflex. Importantly, induction of brain micro-inflammation at specific vessels by cytokine injection was sufficient to establish fatal gastrointestinal failure. Resulting micro-inflammation activated new neural pathway including neurons in paraventricular-nucleus, dorsomedial-nucleus-ofhypothalamus, and also vagal neurons to cause fatal gastrointestinal failure. Suppression of the brain micro-inflammation or blockage of these neural pathways inhibited the gastrointestinal failure. These results demonstrate direct link between brain micro-inflammation and fatal gastrointestinal disease via establishment of a new neural pathway under stress. They further suggest that brain micro-inflammation around specific vessels could be switch to activate new neural pathway(s) to regulate organ homeostasis.
• Changes in right ventricular function assessed by echocardiography in dog models of mild RV pressure overload

  Morita T, Nakamura K, Osuga T, Yokoyama N, Morishita K, Sasaki N, Ohta H, Takiguchi M

  Echocardiography 34 (7) 1040 - 1049 0742-2822 2017/07 [Refereed][Not invited]
   

  BACKGROUND: The assessment of hemodynamic change by echocardiography is clinically useful in patients with pulmonary hypertension. Recently, mild elevation of the mean pulmonary arterial pressure (PAP) has been shown to be associated with increased mortality. However, changes in the echocardiographic indices of right ventricular (RV) function are still unknown. The objective of this study was to validate the relationship between echocardiographic indices of RV function and right heart catheterization variables under a mild RV pressure overload condition. METHODS AND RESULTS: Echocardiography and right heart catheterization were performed in dog models of mild RV pressure overload induced by thromboxane A2 analog (U46619) (n=7). The mean PAP was mildly increased (19.3±1.1 mm Hg), and the cardiac index was decreased. Most echocardiographic indices of RV function were significantly impaired even under a mild RV pressure overload condition. Multivariate analysis revealed that the RV free wall longitudinal strain (RVLS), standard deviation of the time-to-peak longitudinal strain of RV six segments (RV-SD) by speckle-tracking echocardiography, and Tei index were independent echocardiographic predictors of the mean PAP (free wall RVLS, β=-0.60, P<.001; RV-SD, β=0.40, P=.011), pulmonary vascular resistance (free wall RVLS, β=-0.39, P=.020; RV-SD, β=0.47, P=.0086; Tei index, β=0.34, P=.047), and cardiac index (Tei index, β=-0.65, P<.001). CONCLUSIONS: Free wall RVLS, RV-SD, and Tei index are useful for assessing the hemodynamic change under a mild RV pressure overload condition.
• Stimulation of colorectal biopsies from miniature dachshunds with inflammatory colorectal polyps with toll-like receptor ligands: A pilot study

  Yokoyama N, Ohta H, Kagawa Y, Nagata N, Nisa K, Morita T, Osuga T, Sasaki N, Morishita K, Nakamura K, Takiguchi M

  Veterinary Immunology and Immunopathology 188 78 - 83 0165-2427 2017/06 [Refereed][Not invited]
   

  Inflammatory colorectal polyps (ICRPs) in miniature dachshunds (MDs) are a possible novel form of breed-specific canine inflammatory bowel disease (IBD). In this pilot study, we investigated the effects of different Toll like receptor (TLR2, TLR4, TLR5 and TLR9) ligands on pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α) gene expression in ex vivo-cultured colorectal samples from four MDs with ICRPs and four healthy MDs. At baseline, no significant difference in the mRNA expression levels of TLRs and pro-inflammatory cytokines was observed between cases and control MDs. After 4-h incubation, the relative ratios of TNF-α mRNA expression in the TLR2- or TLR4-stimulated colorectal samples, and IL-1β mRNA expression in the TLR9-stimulated colorectal samples form cases showed higher tendency compared with healthy MDs (P<0.05), although statistically not significant. The results of this pilot study using small number of cases indicated that reactivity against TLR2, TLR4 or TLR9 ligand in the production of pro-inflammatory cytokines might be enhanced in the colorectal mucosa of ICRPs. Further research is needed to perform the functional analysis of TLRs in the sole cell population using intestinal epithelial primary culture and the mononuclear cells isolated from colonic mucosa.
• Washout Ratio in the Hepatic Vein Measured by Contrast-Enhanced Ultrasonography to Distinguish Between Inflammatory and Noninflammatory Hepatic Disorders in Dogs

  K. Morishita, A. Hiramoto, A. Michishita, S. Takagi, T. Osuga, S. Y. Lim, K. Nakamura, N. Sasaki, H. Ohta, M. Takiguchi

  JOURNAL OF VETERINARY INTERNAL MEDICINE 31 (3) 770 - 777 1939-1676 2017/05 [Refereed][Not invited]
   

  Background: Perflubutane microbubbles, a second-generation ultrasound contrast agent, are phagocytized by Kupffer cells. This characteristic may be useful to differentiate diffuse hepatic diseases in dogs. Hypothesis/objectives: To determine whether the washout ratio in the hepatic vein (HV) measured by contrast-enhanced ultrasonography (CEUS) can distinguish between inflammatory and noninflammatory hepatic disorders in dogs. Animals: Forty-one client-owned dogs with hepatic disorders including 14 with hepatitis, 7 with primary hypoplasia of the portal vein (PHPV), 9 with congenital portosystemic shunt (cPSS), and 11 with other hepatopathy were enrolled. Six dogs without hepatic disease also were evaluated as healthy controls. Methods: Dogs with hepatic disorders were prospectively included. Contrast-enhanced ultrasonography of the HV was performed for 2 minutes. Washout ratio was defined as the attenuation rate from peak intensity to the intensity at the end of the CEUS study. Results: Washout ratio in the hepatitis group (median, 18.0%; range, 2.0-37.0%) was significantly lower than that of the PHPV (median, 52.2%; range, 11.5-86.3%), cPSS (median, 60.0%; range, 28.6-77.4%), other hepatopathy (median, 70.5%; range, 26.6-88.4%), and normal (median, 78.0%; range, 60.7-91.7%) groups. The area under the receiver operating characteristic curve for hepatitis was 0.960, with a 95% confidence interval (CI) of 0.853-0.990. Washout ratio <= 37.1% resulted in a sensitivity of 100% (95% CI, 78.5-100%) and specificity of 85.2% (95% CI, 67.5-94.1%) for the prediction of hepatitis. Conclusions and Clinical Importance: Washout ratio can distinguish hepatitis from the other noninflammatory disorders with high accuracy. This result might reflect impaired Kupffer cell phagocytosis in dogs with hepatitis.
• Washout Ratio in the Hepatic Vein Measured by Contrast-Enhanced Ultrasonography to Distinguish Between Inflammatory and Noninflammatory Hepatic Disorders in Dogs.

  Morishita K, Hiramoto A, Michishita A, Takagi S, Osuga T, Lim SY, Nakamura K, Sasaki N, Ohta H, Takiguchi M

  Journal of veterinary internal medicine 31 (3) 770 - 777 0891-6640 2017/05 [Refereed][Not invited]
  
• Assessment of contrast-enhanced ultrasonography of the hepatic vein for detection of hemodynamic changes associated with experimentally induced portal hypertension in dogs

  Keitaro Morishita, Akira Hiramoto, Asuka Michishita, Satoshi Takagi, Yuki Hoshino, Takaharu Itami, Sue Yee Lim, Tatsuyuki Osuga, Sayuri Nakamura, Kenji Ochiai, Kensuke Nakamura, Hiroshi Ohta, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  AMERICAN JOURNAL OF VETERINARY RESEARCH 78 (4) 465 - 471 0002-9645 2017/04 [Refereed][Not invited]
   

  OBJECTIVE To assess the use of contrast-enhanced ultrasonography (CEUS) of the hepatic vein for the detection of hemodynamic changes associated with experimentally induced portal hypertension in dogs. ANIMALS 6 healthy Beagles. PROCEDURES A prospective study was conducted. A catheter was surgically placed in the portal vein of each dog. Hypertension was induced by intraportal injection of microspheres (10 to 15 mg/kg) at 5-day intervals via the catheter. Microsphere injections were continued until multiple acquired porto-systemic shunts were created. Portal vein pressure (PVP) was measured through the catheter. Contrast-enhanced ultrasonography was performed before and after establishment of hypertension. Time-intensity curves were generated from the region of interest in the hepatic vein. Perfusion variables measured for statistical analysis were hepatic vein arrival time, time to peak, time to peak phase (TTPP), and washout ratio. The correlation between CEUS variables and PVP was assessed by use of simple regression analysis. RESULTS Time to peak and TTPP were significantly less after induction of portal hypertension. Simple regression analysis revealed a significant negative correlation between TTPP and PVP. CONCLUSIONS AND CLINICAL RELEVANCE CEUS was useful for detecting hemodynamic changes associated with experimentally induced portal hypertension in dogs, which was characterized by a rapid increase in the intensity of the hepatic vein. Furthermore, TTPP, a time-dependent variable, provided useful complementary information for predicting portal hypertension. IMPACT FOR HUMAN MEDICINE Because the method described here induced presinusoidal portal hypertension, these results can be applied to idiopathic portal hypertension in humans.
• Expression of apical junction complex proteins in colorectal mucosa of miniature dachshunds with inflammatory colorectal polyps

  Yokoyama N, Ohta H, Kagawa Y, Leela-Arporn R, Dermlim A, Nisa K, Morita T, Osuga T, Sasaki N, Morishita K, Nakamura K, Takiguchi M

  Journal of Veterinary Medical Science 79 (3) 456 - 463 0916-7250 2017/03 [Refereed][Not invited]
   

  We examine the expression of tight junction and adherence junction proteins in the colorectal mucosa of miniature dachshunds (MDs) with inflammatory colorectal polyps (ICRPs). Colorectal mucosa samples were endoscopically obtained from 8 MDs with ICRPs and 8 control dogs for immunoblotting. Paraffin-embedded tissues of surgically resected inflamed lesions from another 5 MDs with ICRPs and full-thickness colorectal specimens from 5 healthy beagles were obtained for immunohistochemistry. The expression patterns of claudin-1, -2, -3, -4, -5, -7 and -8, E-cadherin and β-catenin were analyzed in the non-inflamed mucosa and inflamed mucosa of ICRPs and colorectal mucosa of control dogs by immunoblotting. The localization of these proteins in the inflamed lesions was analyzed by immunohistochemistry. The expressions of each of claudin, E-cadherin and β-catenin were not significantly different between control dogs and non-inflamed colonic mucosa from MDs with ICRPs. In contrast, only E-cadherin and β-catenin were detected in the inflamed lesions of MDs with ICRPs. By immunohistochemistry, claudin-2, -3, -4, -5 and -7, E-cadherin and β-catenin were expressed in the colorectal epithelium within the inflamed mucosa, but not in granulation tissue. Distributions of claudin-2, -3, -4, -5, and -7, E-cadherin and β-catenin in the colonic epithelium were not different between MDs with ICRPs and control dogs. These results indicated that no significant alteration was detected in several tight junction or adherence junction proteins expression in the colorectal epithelium of ICRPs.
• Contrast-enhanced ultrasonography of the hepatic vein in normal dogs.

  Morishita K, Hiramoto A, Osuga T, Lim SY, Khoirun N, Sasaki N, Nakamura K, Ohta H, Yamasaki M, Takiguchi M

  The Journal of veterinary medical science 78 (12) 1753 - 1758 0916-7250 2017/01 [Refereed][Not invited]
   

  Contrast-enhanced ultrasonography (CEUS) of the hepatic vein can assess intrahepatic hemodynamic changes and has been studied as a noninvasive method to assess the severity of portal hypertension and hepatic fibrosis in humans. However, few reports have described its usefulness in veterinary medicine. The purpose of this study was to characterize CEUS findings of the hepatic vein in normal dogs and assess the repeatability of this method both in a conscious group (n=6) and a sedated group (n=6). Sonazoid® (0.01 ml/kg) was used as a contrast agent, and scanning of the hepatic vein was performed for 2 min. Time-intensity curves were generated from regions of interest in the hepatic vein. Four perfusion parameters were measured for quantitative analysis: hepatic vein arrival time (HVAT), time to peak (TTP), time to peak phase (TTPP) and wash-out ratio (WR). CEUS examinations were performed three times in each dog. The median (range) values of HVAT, TTP, TTPP and WR in the conscious group were 13.5 sec (9-22 sec), 12.5 sec (6-24 sec), 8 sec (6-13 sec) and 78.0% (60.7-91.7%), respectively. Median (range) values of HVAT, TTP, TTPP and WR in the sedated group were 12 sec (8-17 sec), 12.5 sec (9-17 sec), 9 sec (7-13 sec) and 84.1% (63.0-94.4%), respectively. The coefficients of variation of these parameters in the conscious and sedated groups were 7.6-29.7% and 11.8-14.8%, respectively.
• Impact of surgical margins on survival of 37 dogs with massive hepatocellular carcinoma

  A. Matsuyama, S. Takagi, K. Hosoya, Y. Kagawa, K. Nakamura, T. Deguchi, M. Takiguchi

  NEW ZEALAND VETERINARY JOURNAL 65 (5) 227 - 231 0048-0169 2017 [Refereed][Not invited]
   

  AIMS: To compare the survival of dogs with completely resected massive hepatocellular carcinoma (HCC) with that of dogs in which HCC were incompletely excised. METHODS: A retrospective cohort study was conducted. Dogs that underwent surgical excision of massive HCC between November 2006 and April 2015 were included. Dogs that died in the perioperative period or were lost to follow-up within 2 months after surgery were excluded. Data were collected from the medical records and a single pathologist examined all available histology slides to confirm the diagnosis of HCC. Surgical margins were defined as complete if no neoplastic cells were seen at the edge of excised tissues, based on original histopathology reports. Progression-free survival (PFS) and overall survival (OS) were compared between dogs with complete surgical margins (CM) and those with incomplete margins (IM) using a log-rank test. RESULTS: Of the 37 dogs included in the study, 25 were allocated to the CM group and 12 to the IM group. Progressive local disease developed after surgery in three dogs in the CM group and seven dogs in the IM group. Three dogs in the CM group and five dogs in the IM group died due to tumour progression. Median PFS was longer for dogs in the CM group (1,000 (95% CI=562-1,438) days) compared to dogs in the IM group (521 (95% CI=243-799) days; p=0.007). OS was also longer for dogs in the CM group (>1,836 days) compared to those in the IM group (median 765 (95% CI=474-1,056) days; p=0.02). CONCLUSIONS AND CLINICAL RELEVANCE: Compared with complete resection, incomplete resection decreased PFS and OS in dogs with massive HCC. Dogs with incompletely excised HCC should be closely monitored for local recurrence, although median OS was >2 years following incomplete excision. Further prospective studies are warranted to confirm these findings.
• Urinary exosome-derived microRNAs reflecting the changes of renal function and histopathology in dogs

  Osamu Ichii, Hiroshi Ohta, Taro Horino, Teppei Nakamura, Marina Hosotani, Tatsuya Mizoguchi, Keitaro Morishita, Kensuke Nakamura, Yuki Hoshino, Satoshi Takagi, Noboru Sasaki, Mitsuyoshi Takiguchi, Ryo Sato, Kazuhisa Oyamada, Yasuhiro Kon

  SCIENTIFIC REPORTS 7 40340  2045-2322 2017/01 [Refereed][Not invited]
   

  MicroRNAs act as post-transcriptional regulators, and urinary exosome (UExo)-derived microRNAs may be used as biomarkers. Herein, we screened for UExo-derived microRNAs reflecting kidney disease (KD) status in dogs. Examined dogs were divided into healthy kidney control (HC) and KD groups according to renal dysfunction. We confirmed the appearance of UExo having irregular globe-shapes in a dog by immunoblot detection of the exosome markers, TSG101 and CD9. Based on our previous data using KD model mice and the data obtained herein by next generation sequencing of UExo-derived microRNAs in dogs, miR-26a, miR-146a, miR-486, miR-21a, and miR-10a/b were selected as candidate microRNAs. In particular, UExo-derived miR-26a and miR-10a/b were significantly decreased in KD dogs, and miR-26a levels negatively correlated with deteriorated renal function compared to the other miRNAs. UExo-derived miR-21a levels corrected or not to that of internal control microRNAs in UExo, miR-26a and miR-191, significantly increased with renal dysfunction. In kidney tissues, the decrease of miR-26a and miR-10a/ b in the glomerulus and miR-10b in the tubulointerstitium negatively correlated with deteriorated renal function and histopathology. Increased miR-21a in the tubulointerstitium rather than in the glomerulus correlated with deteriorated renal histopathology. In conclusion, microRNAs reflecting the changes in renal function and histopathology in dogs were identified in this study.
• Serial measurement of pancreatic lipase immunoreactivity concentration in dogs with immune-mediated disease treated with prednisolone

  Ohta H, Morita T, Yokoyama N, Osuga T, Sasaki N, Morishita K, Nakamura K, Takiguchi M

  Journal of Small Animal Practice 58 (6) 342 - 347 0022-4510 2017 [Refereed][Not invited]
   

  Objectives: In this pilot study, serum canine pancreatic lipase immunoreactivity was measured repeatedly in dogs with various immune-mediated diseases that were treated with immunosuppressive doses of prednisolone. Methods: Ten client-owned dogs with newly diagnosed immune-mediated disease that had normal canine pancreatic lipase immunoreactivity concentrations (≤200 µg/l) were treated with 2 to 2.2 mg/kg prednisolone orally once daily as the initial treatment. Serum samples were obtained from each of the dogs prior to treatment and at 1- to 4-week intervals during immunosuppressive treatment. The highest canine pancreatic lipase immunoreactivity concentration detected during immunosuppressive treatment was defined as the peak canine pancreatic lipase immunoreactivity. Results: Peak canine pancreatic lipase immunoreactivity concentrations were classified as normal in two dogs, questionable (201 to 399 µg/l) in three dogs, and abnormal (≥400 µg/l) in five dogs. Peak canine pancreatic lipase immunoreactivity concentrations were significantly higher than baseline canine pancreatic lipase immunoreactivity concentrations but there was no evidence of clinical pancreatitis. Clinical Significance: It remains unclear whether the five of 10 dogs with elevated canine pancreatic lipase immunoreactivity during prednisone treatment had subclinical pancreatitis or whether the abnormal results were a consequence of prednisolone administration.
• Localization of Toll-like Receptor (TLR) 2 and TLR4 mRNA in the Colorectal Mucosa of Miniature Dachshunds with Inflammatory Colorectal Polyps

  Yokoyama N, Ohta H, Yamazaki J, Kagawa Y, Ichii O, Khoirun N, Morita T, Osuga T, Lim S.Y, Sasaki N, Morishita K, Nakamura K, Takiguchi M

  Journal of Comparative Pathology 156 (2-3) 183 - 190 0021-9975 2017 [Refereed][Not invited]
   

  Inflammatory colorectal polyps (ICRPs) are characterized by the formation of multiple or solitary polyps with marked neutrophil infiltration in the colorectal area, and are speculated to be a novel form of breed-specific canine idiopathic inflammatory bowel disease (IBD). In human IBD, toll-like receptor (TLR) 2 and TLR4 have been reported to be involved in the pathogenesis of the disease. The aim of this study was to evaluate the expression of TLR2 and TLR4 mRNA in the colorectal mucosa of dogs with ICRPs by in-situ hybridization using an RNAscope assay. Samples of inflamed colorectal mucosa (n = 5) and non-inflamed mucosa (n = 5) from miniature dachshunds (MDs) with ICRPs and colonic mucosa from healthy beagles (n = 5) were examined. TLR2 and TLR4 hybridization signals were localized to the colorectal epithelium, inflammatory cells and fibroblasts in the inflamed colorectal mucosa of affected dogs. The signals were significantly greater in inflamed colorectal epithelium compared with non-inflamed epithelium of MDs with ICRPs and healthy beagles (P <0.05). These results suggest that increased expression of TLR2 and TLR4 mRNA in the inflamed colorectal mucosa results from not only inflammatory cell infiltration, but also the upregulation of TLR2 and TLR4 mRNA in the colonic epithelium.
• Left Atrial Strain at Different Stages of Myxomatous Mitral Valve Disease in Dogs

  Nakamura K, Kawamoto S, Osuga T, Morita T, Sasaki N, Morishita K, Ohta H, Takiguchi M

  Journal of Veterinary Internal Medicine 31 (2) 316 - 325 2017 [Refereed][Not invited]
  
• The repeatability and characteristics of right ventricular longitudinal strain imaging by speckle-tracking echocardiography in healthy dogs

  Morita T, Nakamura K, Osuga T, Yokoyama N, Khoirun N, Morishita K, Sasaki N, Ohta H, Takiguchi M

  Journal of Veterinary Cardiology 19 (4) 351 - 362 1760-2734 2017 [Refereed][Not invited]
   

  Introduction To assess the repeatability and characteristics of echocardiographic indices of the right ventricular (RV) function derived from speckle-tracking echocardiography. Animals Fourteen laboratory Beagles and 103 privately owned dogs without cardiac disease were involved in this study. Materials and methods Right ventricular longitudinal strain, strain rate, and a strain-related index for assessing RV dyssynchrony derived from speckle-tracking echocardiography were obtained by two different observers using five Beagles. Within-day, between-day, and interobserver coefficients of variation and the intraclass correlation coefficient of speckle-tracking echocardiography indices were determined. Both speckle-tracking echocardiography and conventional indices of RV function, including the peak velocity of systolic tricuspid annular motion, tricuspid annulus plane systolic excursion, fractional area change, and the Tei index, were obtained from 14 Beagles and 103 privately owned dogs. Relationships between echocardiographic indices and the body weight, heart rate, age, and sex were estimated by regression analysis. Results Speckle-tracking echocardiographic indices showed good within-day repeatability, between-day and interobserver repeatability were moderate to good. In large dogs, RV longitudinal strain, strain rate, and fractional area change were significantly decreased, while the index of RV dyssynchrony, systolic tricuspid annular motion, tricuspid annulus plane systolic excursion, and the Tei index were increased. All speckle-tracking and conventional echocardiographic indices were correlated with the body weight. Discussion and conclusions The speckle-tracking echocardiography indices were highly repeatable and body weight affected speckle-tracking echocardiography indices in dogs. Further studies are needed to apply speckle-tracking echocardiography indices in dogs with cardiac disease.
• Molecular cloning, phylogenetic analysis and heat shock response of Babesia gibsoni heat shock protein 90.

  Masahiro Yamasaki, Yoshihiro Tsuboi, Yusuke Taniyama, Naohiro Uchida, Reeko Sato, Kensuke Nakamura, Hiroshi Ohta, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 78 (8) 1355 - 60 0916-7250 2016/09/01 [Refereed][Not invited]
   

  The Babesia gibsoni heat shock protein 90 (BgHSP90) gene was cloned and sequenced. The length of the gene was 2,610 bp with two introns. This gene was amplified from cDNA corresponding to full length coding sequence (CDS) with an open reading frame of 2,148 bp. A phylogenetic analysis of the CDS of HSP90 gene showed that B. gibsoni was most closely related to B. bovis and Babesia sp. BQ1/Lintan and lies within a phylogenetic cluster of protozoa. Moreover, mRNA transcription profile for BgHSP90 exposed to high temperature were examined by quantitative real-time reverse transcription-polymerase chain reaction. BgHSP90 levels were elevated when the parasites were incubated at 43°C for 1 hr.
• Effects of experimental cardiac volume loading on left atrial phasic function in healthy dogs

  Osuga T, Nakamura K, Morita T, Nisa K, Yokoyama N, Sasaki N, Morishita K, Ohta H, Takiguchi M

  American Journal of Veterinary Research 77 (9) 952 - 960 0002-9645 2016/09 [Refereed][Not invited]
   

  OBJECTIVE To elucidate the relationship between acute volume overload and left atrial phasic function in healthy dogs. ANIMALS 6 healthy Beagles. PROCEDURES Dogs were anesthetized. A Swan-Ganz catheter was placed to measure mean pulmonary capillary wedge pressure (PCWP). Cardiac preload was increased by IV infusion with lactated Ringer solution at 150 mL/kg/h for 90 minutes. Transthoracic echocardiography was performed before (baseline) and at 15, 30, 45, 60, 75, and 90 minutes after volume loading began. At each echocardiographic assessment point, apical 4-chamber images were recorded and analyzed to derive time-left atrial area curves. Left atrial total (for reservoir function), passive (for conduit function), and active (for booster-pump function) fractional area changes were calculated from the curves. RESULTS Volume overload resulted in a significant increase from baseline in PCWP from 15 to 90 minutes after volume loading began. All fractional area changes at 15 to 90 minutes were significantly increased from baseline. In multiple regression analysis, quadratic regression models were better fitted to the relationships between PCWP and each of the total and active fractional area changes than were linear regression models. A linear regression model was better fitted to the relationship between PCWP and passive fractional area change. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that left atrial phasic function assessed on the basis of left atrial phasic areas was enhanced during experimental cardiac volume loading in healthy dogs. The effect of volume load should be considered when evaluating left atrial phasic function by indices derived from left atrial phasic sizes.
• Hypercalcemia in a Dog with Chronic Ingestion of Maxacalcitol Ointment.

  Nakamura K, Tohyama N, Yamasaki M, Ohta H, Morishita K, Takiguchi M

  Journal of the American Animal Hospital Association 52 (4) 256 - 258 0587-2871 2016/07 [Refereed][Not invited]
  
• Repeatability and reproducibility of right ventricular Tei index valves derived from three echocardiographic methods for evaluation of cardiac function in dogs

  Morita T, Nakamura K, Osuga T, Lim S.Y, Yokoyama N, Morishita K, Ohta H, Takiguchi M

  American Journal of Veterinary Research 77 (7) 715 - 720 0002-9645 2016/07 [Refereed][Not invited]
   

  OBJECTIVE To evaluate repeatability and reproducibility of right ventricular Tei index (RTX) values derived from dual pulsed-wave Doppler, conventional pulsed-wave Doppler, and tissue Doppler echocardiography and to investigate relationships and repeatability among the 3 methods in healthy dogs. ANIMALS 6 healthy adult Beagles. PROCEDURE Echocardiography was performed on each dog on different days for 2 weeks (3 times/d) by 2 echocardiographers. Intraobserver within- and between-day and interobserver coefficients of variation (CVs) and intraclass correlation coefficients (ICCs) for RTXs derived from dual pulse-waved Doppler (RTXDPD), conventional pulsed-wave Doppler (RTXPD), and tissue Doppler (RTXTD) methods were determined. Degrees of agreement among RTX values derived from the 3 methods were assessed by modified Bland-Altman analysis. RESULTS Least squares mean (95% confidence interval) RTXTD was 0.50 (0.46 to 0.54), which was significantly higher than that for RTXDPD (0.27 [0.23 to 0.31]) and RTXPD (0.25 [0.21 to 0.29]). Agreement between RTXDPD and RTXPD was good (bias [mean difference], 0.04 [95% confidence interval, –0.03 to 0.10]). The RTXDPD had high within-day (CV, 6.1; ICC, 0.77) and interobserver (CV, 3.5; ICC, 0.83) repeatability, but between-day repeatability was not high. The RTXTD had high within-day repeatability (CV, 6.0; ICC, 0.80), but between-day and interobserver repeatability were not high. Within-day, between-day, and interobserver repeatability of RTXPD were not high. CONCLUSIONS AND CLINICAL RELEVANCE RTXDPD measurement was a repeatable and reproducible method of cardiac evaluation in healthy dogs. The RTXTD values were significantly higher than the RTXDPD and RTXPD values; therefore, RTX values derived from different echocardiographic methods should be interpreted with caution.
• Prognostic Value of Right Ventricular Tei Index in Dogs with Myxomatous Mitral Valvular Heart Disease

  Nakamura K, Morita T, Osuga T, Morishita K, Sasaki N, Ohta H, Takiguchi M

  Journal of Veterinary Internal Medicine Wiley-Blackwell 30 (1) 69 - 75 0891-6640 2016 [Refereed][Not invited]
   

  Background: The right ventricular (RV) Tei index (TX) has a significant correlation with the severity of pulmonary hypertension. However, the role of RV dysfunction in dogs with myxomatous mitral valvular heart disease (MMVD) has not been addressed. Objectives: To investigate the correlation between right ventricular Tei-index (RVTX) and the prognosis for dogs with MMVD. Animals: Thirty client-owned dogs with MMVD. Methods: Clinical cohort study. Dogs were divided into two groups on the basis of the onset of cardiac-related death within 1 year of the first echocardiographic examination. Physical examination and echocardiographic variables were compared between the groups. Receiver operating characteristic (ROC) curves and multivariate logistic analysis were used to assess the comparative accuracy when identifying dogs with cardiac-related death. Results: The highest accuracy was obtained for RVTX with an area under the ROC curve (AUC) of 0.95 (95% confidence interval [CI] 0.81-0.99) followed by the left atrial to aortic root ratio with an AUC of 0.91 (95% CI 0.74-0.98), peak early diastolic mitral inflow velocity with an AUC of 0.84 (95% CI 0.64-0.94), and Doppler estimates of systolic pulmonary artery pressure with an AUC of 0.84 (95% CI 0.61-0.95). According to the multivariate logistic regression analysis, RVTX was the only independent correlate of cardiac-related death within 1 year. Conclusions and Clinical Importance: Right ventricular Tei-index has a strong correlation with the prognosis for dogs with MMVD. The most significant independent predictor of death was RVTX in this study.
• Effects of various cardiovascular drugs on indices obtained with two-dimensional speckle tracking echocardiography of the left atrium and time-left atrial area curve analysis in healthy dogs

  Tatsuyuki Osuga, Kensuke Nakamura, Tomoya Morita, Sue Yee Lim, Nozomu Yokoyama, Keitaro Morishita, Hiroshi Ohta, Mitsuyoshi Takiguchi

  AMERICAN JOURNAL OF VETERINARY RESEARCH 76 (8) 702 - 709 0002-9645 2015/08 [Refereed][Not invited]
   

  OBJECTIVE To evaluate the effects of dobutamine, esmolol, milrinone, and phenylephrine on left atrial phasic function of healthy dogs. ANIMALS 9 healthy Beagles. PROCEDURES Following sedation with propofol on each of 4 experimental days, dogs were administered a constant rate infusion of dobutamine (5 mu g/kg/min), esmolol (500 mu g/kg/min), milrinone (25 mu g/kg, IV bolus, followed by 0.5 mu g/kg/min), or phenylephrine (2 mu g/kg/min). There was at least a 14-day interval between experimental days. Each drug was administered to 6 dogs. Conventional and 2-D speckle tracking echocardiography were performed before (baseline) and after administration of the cardiovascular drug, and time-left atrial area curves were derived to calculate indices for left atrial reservoir, conduit, and booster pump functions (left atrial phasic function) and left ventricular contractility and lusitropy. RESULTS Compared with baseline values, indices for left atrial reservoir and booster pump functions and left ventricular contractility and lusitropy were significantly increased following dobutamine administration; indices for left atrial phasic function and left ventricular lusitropy were changed insignificantly, and indices for left ventricular contractility were significantly impaired following esmolol administration; indices for left atrial phasic function and left ventricular relaxation were changed insignificantly, and indices for left ventricular systolic function were significantly augmented following milrinone administration; and indices for left atrial phasic function and left ventricular lusitropy were changed insignificantly, and indices of ventricular contractility were significantly impaired following phenylephrine administration. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that, following administration of dobutamine, esmolol, milrinone, or phenylephrine to healthy dogs, left atrial phasic function indices were fairly stable and did not parallel changes in left ventricular function indices.
• Quantitative contrast-enhanced ultrasonographic assessment of naturally occurring pancreatitis in dogs.

  Lim SY, Nakamura K, Morishita K, Sasaki N, Murakami M, Osuga T, Yokoyama N, Ohta H, Yamasaki M, Takiguchi M

  Journal of veterinary internal medicine / American College of Veterinary Internal Medicine 29 (1) 71 - 78 0891-6640 2015/01 [Refereed][Not invited]
   

  Background: Quantitative contrast-enhanced ultrasonography (CEUS) can detect pancreatic perfusion changes in experimentally induced canine pancreatitis. However, its usefulness in detecting perfusion changes in naturally occurring pancreatitis is unclear. Hypothesis/Objectives: To determine the feasibility of using CEUS to detect pancreatic and duodenal perfusion changes in naturally occurring canine pancreatitis. Animals: Twenty-three client-owned dogs with pancreatitis, 12 healthy control dogs. Methods: Dogs diagnosed with pancreatitis were prospectively included. CEUS of the pancreas and duodenum were performed. Time-intensity curves were created from regions of interest in the pancreas and duodenum. Five perfusion parameters were obtained for statistical analyses: time to initial up-slope, peak time (Tp), time to wash-out (TTW), peak intensity (PI), and area under the curve (AUC). Results: For the pancreas, Tp of the pancreatitis group was prolonged when compared to controls (62 ± 11 seconds versus 39 ± 13 seconds; P < .001). TTW also was prolonged but not significantly (268 ± 69 seconds versus 228 ± 47 seconds; P = .47). PI and AUC were increased when compared to controls (95 ± 15 versus 78 ± 13 MPV; P = .009 and 14,900 ± 3,400 versus 11,000 ± 2,800 MPV*s; P = .013, respectively). For the duodenum, PI and AUC were significantly increased in the pancreatitis group when compared to controls. Conclusions and Clinical Importance: Contrast-enhanced ultrasonography can detect pancreatic perfusion changes in naturally occurring canine pancreatitis characterized by delayed peak with prolonged hyperechoic enhancement of the pancreas on CEUS. Additionally, duodenal perfusion changes secondary to pancreatitis were observed.
• Vitamin D Status in Different Stages of Disease Severity in Dogs with Chronic Valvular Heart Disease

  Osuga T, Nakamura K, Morita T, Lim S.Y, Nisa K, Yokoyama N, Sasaki N, Morishita K, Ohta H, Takiguchi M

  Journal of Veterinary Internal Medicine 29 (6) 1518 - 1523 0891-6640 2015 [Refereed][Not invited]
   

  Background: In humans with heart disease, vitamin D deficiency is associated with disease progression and a poor prognosis. A recent study showed that serum 25-hydroxyvitamin D [25(OH)D] concentration, the hallmark of vitamin D status, was lower in dogs with heart failure than in normal dogs, and a low concentration was associated with poor outcome in dogs with heart failure. Objectives: To elucidate the vitamin D status of dogs with chronic valvular heart disease (CVHD) at different stages of disease severity. Animals: Forty-three client-owned dogs with CVHD. Methods: In this cross-sectional study, dogs were divided into 3 groups (14 dogs in Stage B1, 17 dogs in Stage B2, and 12 dogs in Stage C/D) according to ACVIM guidelines. Dogs underwent clinical examination including echocardiography. Serum 25(OH)D concentrations were measured in each dog. Results: Serum 25(OH)D concentration was significantly lower in Stage B2 (median, 33.2 nmol/L; range, 4.9-171.7 nmol/L) and C/D (13.1 nmol/L; 4.9-58.1 nmol/L) than in Stage B1 (52.5 nmol/L; 33.5-178.0 nmol/L) and was not significantly different between Stage B2 and Stage C/D. Among clinical variables, there were significant negative correlations between 25(OH)D concentration and both left atrial-to-aortic root ratio and left ventricular end-diastolic diameter normalized for body weight. Conclusions and Clinical Importance: These results indicate that vitamin D status is associated with the degree of cardiac remodeling, and the serum 25(OH)D concentration begins to decrease before the onset of heart failure in dogs with CVHD.
• CONTRAST-ENHANCED ULTRASONOGRAPHIC FINDINGS IN THREE DOGS WITH PANCREATIC INSULINOMA

  Kensuke Nakamura, Sue-Yee Lim, Kenji Ochiai, Masahiro Yamasaki, Hiroshi Ohta, Keitaro Morishita, Satoshi Takagi, Mitsuyoshi Takiguchi

  VETERINARY RADIOLOGY & ULTRASOUND 56 (1) 55 - 62 1058-8183 2015/01 [Refereed][Not invited]
   

  Abdominal ultrasonography is one of the most common diagnostic imaging modalities used for dogs with suspected insulinoma; however, pancreatic masses are clearly identified in fewer than half of affected dogs and benign pancreatic nodules can be difficult to differentiate from malignant ones. The purpose of this prospective study was to describe contrast-enhanced ultrasonography (CEUS) characteristics of confirmed pancreatic insulinoma in a group of dogs. Inclusion criteria were as follows: (1) repeated hypoglycemia (blood glucose levels <60 mg/dl, twice or more); (2) elevated blood insulin levels with hypoglycemia; (3) pancreatic nodules detected with conventional ultrasonography; and (4) histological confirmation of pancreatic islet cell carcinoma. Immediately following conventional ultrasonography of the entire abdomen, CEUS of the pancreatic nodule and adjacent parenchyma was performed using contrast-specific technology pulse inversion imaging and perflubutane microbubble contrast agent. Three dogs met inclusion criteria. Pancreatic nodules in all the three dogs became more clearly demarcated after injection of the contrast agent. Each nodule showed different enhancement patterns: markedly hyperechoic for 5 s, slightly hyperechoic for 1 s, and clearly hypoechoic for over 30 s. These results were not in complete agreement with previously reported CEUS findings in human patients with insulinoma. All nodules were surgically resected and histopathologically confirmed as malignant insulinomas. Findings from the current study indicated that contrast-enhanced ultrasound may help to increase conspicuity of pancreatic insulinomas in dogs and that enhancement characteristics may be more variable in dogs than in humans. (C) 2014 American College of Veterinary Radiology.
• Lesion Type Classification by Applying Machine-Learning Technique to Contrast-Enhanced Ultrasound Images

  Kazuya Takagi, Satoshi Kondo, Kensuke Nakamura, Mitsuyoshi Takiguchi

  IEICE TRANSACTIONS ON INFORMATION AND SYSTEMS E97D (11) 2947 - 2954 1745-1361 2014/11 [Refereed][Not invited]
   

  One of the major applications of contrast-enhanced ultrasound (CEUS) is lesion classification. After contrast agents are administered, it is possible to identify a lesion type from its enhancement pattern. However, CEUS image reading is not easy because there are various types of enhancement patterns even for the same type of lesion, and clear classification criteria have not yet been defined. Some studies have used conventional time intensity curves (TICs), which show the vessel dynamics of a lesion. It is possible to predict lesion type from the TIC parameters, such as the coefficients obtained by curve fitting, peak intensity, flow rate and time to peak. However, these parameters are not always provide sufficient accuracy. In this paper, we prepare 1D Haar-like features which describe intensity changes in a TIC and adopt the Adaboost machine learning technique, which eases understanding of which features are useful. Hyperparameters of weak classifiers, e.g., the step size of a Haar-like filter length and threshold for output of the filter, are optimized by searching for those parameters that give the best accuracy. We evaluate the proposed method using 36 focal splenic lesions in canines 16 of which were benign and 20 malignant. The accuracies were 91.7% (33/36) when inspected by an experienced veterinarian, 75.0% (27/36) by linear discriminant analysis (LDA) using conventional three TIC parameters: time to peak, area under curve and peak intensity, and 91.7% (33/36) using our proposed method. McNemar testing shows the p-value to be less than 0.05 between the proposed method and LDA. This result shows the statistical significance of differences between the proposed method and the conventional TIC analysis method using LDA.
• Evaluation of Selected Cytokine Gene Expression in Colonic Mucosa from Dogs with Idiopathic Lymphocytic-plasmacytic Colitis

  Yu Tamura, Hiroshi Ohta, Nozomu Yokoyama, Sue Yee Lim, Tatsuyuki Osuga, Keitaro Morishita, Kensuke Nakamura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  JOURNAL OF VETERINARY MEDICAL SCIENCE 76 (10) 1407 - 1410 0916-7250 2014/10 [Refereed][Not invited]
   

  Lymphocytic-plasmacytic colitis (LPC) is a common form of inflammatory bowel disease (IBD) affecting the canine large intestine. Cytokines are thought to be involved in the pathogenesis of IBD. However, to date, few studies have investigated cytokine mRNA expression in dogs with LPC. In this study, we investigated mRNA transcription levels of T helper cell cytokines, such as IFN-gamma, IL-4, IL-17 and IL-10 and pro-inflammatory cytokines, such as IL-1 beta, IL-6, TNF-alpha., IL-8, IL-12 and IL-23, in colonic mucosa from LPC dogs by quantitative real-time RT-PCR. No significant differences were detected in cytokine mRNA expressions between dogs with LPC and controls, except for IL-23p19. Dogs with LPC failed to express a predominant cytokine profile in inflamed colonic mucosa as opposed to human IBD.
• In vitro and in vivo safety and efficacy Studies of amphotericin B on Babesia gibsoni

  Masahiro Yamasaki, Eriko Harada, Yu Tamura, Sue Yee Lim, Tatsuyuki Ohsuga, Nozomu Yokoyama, Keitaro Morishita, Kensuke Nakamura, Hiroshi Ohta, Mitsuyoshi Takiguchi

  VETERINARY PARASITOLOGY 205 (3-4) 424 - 433 0304-4017 2014/10 [Refereed][Not invited]
   

  Babesia gibsoni is a causative pathogen of canine babesiosis, which is commonly treated with anti-babesial drugs; however, the development of novel, more effective anti-babesial drugs is necessary because the currently used drugs cannot remove the parasites from dogs. Therefore we investigated the anti-babesial effect of amphotericin B (AmB), a membrane-active polyene macrolide antibiotic. The interaction of such compounds with sterols in bilayer cell membranes can lead to cell damage and ultimately cell lysis. AmB exhibits in vitro activity against B. gibsoni in normal canine erythrocytes within 12 h. We also studied liposomal AmB (L-AmB), a liposomal formulation of AmB that required a longer incubation period to reduce the number of parasites. However, L-AmB completely inhibited the invasion of free parasites into erythrocytes. These results indicated that free parasites failed to invade erythrocytes in the presence of L-AmB. Both AmB and L-AmB induced mild hemolysis of erythrocytes. Moreover, the methemoglobin level and the turbidity index of erythrocytes were significantly increased when erythrocytes were incubated with AmB, suggesting that AmB induced oxidative damage in erythrocytes. Finally, the anti-babesial activity of AmB in vivo was observed. When experimentally B. gibsoni-infected dogs were administered 0.5 and 1 mg/kg AmB by the intravenous route, the number of parasites decreased; however, recurrence of parasitemia was observed, indicating that AmB did not eliminate parasites completely. Blood urea nitrogen and creatinine of dogs were abnormally elevated after the administration of 1 mg/kg AmB. These results indicate that AmB has in vivo activity against B. gibsoni; however, it does not eliminate parasites from infected dogs and affects kidney function at a high dose. (C) 2014 Elsevier B.V. All rights reserved.
• Case report: Protein-losing enteropathy caused by Mesocestoides vogae (syn. M. corti) in a dog

  Yu Tamura, Hiroshi Ohta, Takuya Kashiide, Jun Matsumoto, Tatsuya Sakurai, Nozomu Yokoyama, Keitaro Morishita, Kensuke Nakamura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  VETERINARY PARASITOLOGY 205 (1-2) 412 - 415 0304-4017 2014/09 [Refereed][Not invited]
   

  An eight-year-old, neutered, female Shetland Sheepdog presented with a 6-week history of small intestinal diarrhea. Regenerative anemia, hypoproteinemia, and an increased plasma C-reactive protein concentration were detected On blood examination. Fecal examination and abdominal radiography were unremarkable. Abdominal ultrasonography showed diffusely hyperechoic mucosa in the small intestine. Gastroduodenoscopy, performed under general anesthesia, revealed mucosal edema and increased granularity in the duodenum and jejunum. Histopathological examination of the endoscopically biopsied small intestinal mucosa revealed tapeworm infection. A single administration of a combined anthelmintic drug (5 mg/kg praziquantel, 14.4 mg/kg pyrantel pamoate, and 15 mg/kg febantel) was successful for deworming, and the dog fully recovered. The parasites were removed from stored frozen duodenal mucosa and morphologically identified as Mesocestoides sp. immature adult worms. Mitochondrial (mt) 12S rDNA and mt cytochrome c oxide subunit I genes were amplified from the parasites. DNA sequence analysis showed that the genes shared 100% identity with those of reported M. vogae (syn. M. corti). This is the first reported case of protein-losing enteropathy caused by M. vogae in a dog. (C) 2014 Elsevier B.V. All rights reserved.
• First detection of Echinococcus multilocularis infection in two species of nonhuman primates raised in a zoo: A fatal case in Cercopithecus diana and a strongly suspected case of spontaneous recovery in Macaca nigra

  Kimiaki Yamano, Hirokazu Kouguchi, Kohji Uraguchi, Takeshi Mukai, Chikako Shibata, Hideaki Yamamoto, Noboru Takaesu, Masaki Ito, Yoshinori Makino, Mitsuyoshi Takiguchi, Kinpei Yagi

  PARASITOLOGY INTERNATIONAL 63 (4) 621 - 626 1383-5769 2014/08 [Refereed][Not invited]
   

  The causative parasite of alveolar echinococcosis, Echinococcus multilocularis, maintains its life cycle between red foxes (Vulpes vulples, the definitive hosts) and voles (the intermediate hosts) in Hokkaido, Japan. Primates, including humans, and some other mammal species can be infected by the accidental ingestion of eggs in the feces of red foxes. In August 2011, a 6-year-old zoo-raised female Diana monkey (Cercopithecus diana) died from alveolar echinococcosis. E. multilocularis infection was confirmed by histopathological examination and detection of the E. multilocularis DNA by polymerase chain reaction (PCR). A field survey in the zoo showed that fox intrusion was common, and serodiagnosis of various nonhuman primates using western blotting detected a case of a 14-year-old female Celebes crested macaque (Macaca nigra) that was weakly positive for E. multilocularis. Computed tomography revealed only one small calcified lesion (approximately 8 mm) in the macaque's liver, and both western blotting and enzyme-linked immunosorbent assay (ELISA) showed a gradual decline of antibody titer. These findings strongly suggest that the animal had recovered spontaneously. Until this study, spontaneous recovery from E. multilocularis infection in a nonhuman primate had never been reported. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
• Expression of apical junction complex proteins in duodenal mucosa of dogs with inflammatory bowel disease

  Hiroshi Ohta, Yuji Sunden, Nozomu Yokoyama, Tatsuyuki Osuga, Sue Yee Lim, Yu Tamura, Keitaro Morishita, Kensuke Nakamura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  AMERICAN JOURNAL OF VETERINARY RESEARCH 75 (8) 746 - 751 0002-9645 2014/08 [Refereed][Not invited]
   

  Objective-To determine the expression of tight junction and adherens junction proteins in duodenal mucosa samples of dogs with inflammatory bowel disease (IBD). Animals-12 dogs with IBD and 6 healthy control Beagles. Procedures-Duodenal mucosa biopsy samples were endoscopically obtained from dogs with IBD and healthy control Beagles. The expression of claudin-1, -2, -3, -4, -5, -7, and -8; E-cadherin; and beta-catenin in the duodenal mucosa samples was determined by means of immunoblotting. The subcellular localization of E-cadherin in the duodenal mucosa samples was determined with immunofluorescence microscopy. Results-The expression of each claudin and beta-catenin was not significantly different between control dogs and dogs with IBD. However, expression of E-cadherin was significantly lower in duodenal mucosa samples of dogs with IBD than it was in samples obtained from healthy control dogs. Results of immunofluorescence microscopy indicated decreased intensity of E-cadherin labeling in the tips of villi in duodenal mucosa samples obtained from 6 dogs with IBD, compared with staining intensity for other dogs. Conclusions and Clinical Relevance-Results of this study indicated expression of claudin-1, -2, -3, -4, -5, -7, and -8 and beta-catenin was not significantly different between duodenal mucosa samples obtained from control dogs and those obtained from dogs with IBD. However, E-cadherin expression was significantly lower in the villus epithelium in duodenal mucosa samples obtained from dogs with IBD versus samples obtained from control dogs, which suggested that decreased expression of that protein has a role in the pathogenesis of IBD in dogs.
• Qualitative and Quantitative Contrast-Enhanced Ultrasonographic Assessment of Cerulein-Induced Acute Pancreatitis in Dogs

  S. Y. Lim, K. Nakamura, K. Morishita, N. Sasaki, M. Murakami, T. Osuga, H. Ohta, M. Yamasaki, M. Takiguchi

  JOURNAL OF VETERINARY INTERNAL MEDICINE 28 (2) 496 - 503 0891-6640 2014/03 [Refereed][Not invited]
   

  Background Acute pancreatitis (AP) is the most common disease of the canine exocrine pancreas, and accurate noninvasive diagnosis is challenging. Hypothesis/Objectives To determine the feasibility of using quantitative contrast-enhanced ultrasonography (CEUS) to detect pancreatic perfusional changes in cerulein-induced AP in dogs. Animals Six adult female Beagles. Methods Each dog received 2hours of IV infusion with 7.5 mu g/kg/h of cerulein diluted in saline. As control, all dogs received 2hours of IV infusion of saline 2weeks before cerulein infusion. CEUS of the pancreas and duodenum were performed before (0hour), and at 2, 4, 6, and 12hours after saline and cerulein infusion. Time-intensity curves were created from regions of interest in the pancreas and duodenum. Five perfusional parameters were measured for statistical analysis: time to initial up-slope, peak time, time to wash-out, peak intensity (PI), and area under the curve (AUC). Results In cerulein-induced AP, pancreatic PI increased at 2 and 4hours when compared to 0hour, and at 2, 4, and 6hours when compared to control. AUC increased at 4hours when compared to 0hour, and at 2 and 4hours when compared to control. Time to wash-out was prolonged at 4hours when compared to control. For saline control, peak time was faster at 2hours when compared to 0hour. Conclusions and Clinical Importance CEUS parameters PI and AUC can provide useful information in differentiating acute pancreatitis from normal pancreas. Cerulein-induced AP was characterized by prolonged hyperechoic enhancement on CEUS.
• CD4(+) T Cell Cytokine Gene and Protein Expression in Duodenal Mucosa of Dogs with Inflammatory Bowel Disease

  Hiroshi Ohta, Kanae Takada, Yuji Sunden, Yu Tamura, Tatsuyuki Osuga, Sue Yee Lim, Masahiro Murakami, Noboru Sasaki, Bandula Kumara Wickramasekara Rajapakshage, Kensuke Nakamura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  JOURNAL OF VETERINARY MEDICAL SCIENCE 76 (3) 409 - 414 0916-7250 2014/03 [Refereed][Not invited]
   

  Inflammatory bowel disease (IBD) is a common cause of chronic gastrointestinal signs in dogs. In humans, T helper cells have important roles in the pathogenesis of IBD. In contrast, no specific involvement of a distinct T cell subset has been described in canine IBD. The present study evaluated the gene and protein expression of cytokines of T cell subsets in duodenal mucosa from dogs with IBD. Relative quantification of interleukin (IL)-17A, interferon (IFN)-gamma, IL-4 and IL-10 mRNA transcription was performed using duodenal mucosa from 27 IBD dogs and 8 controls. Duodenal mucosal IL-17A, IFN-gamma and IL-10 protein levels were determined by ELISA in 15 IBD dogs and 8 controls. There was no significant difference in each cytokines mRNA transcription level between groups. There was no significant difference in IL-17A, IFN-gamma and IL-10 protein expression levels between groups. Thus, there is no clear evidence for the involvement of distinct Th cytokine in the pathogenesis of canine IBD.
• Ultrasound image-guided therapy enhances antitumor effect of cisplatin.

  Noboru Sasaki, Nobuki Kudo, Kensuke Nakamura, Sue Yee Lim, Masahiro Murakami, W R Bandula Kumara, Yu Tamura, Hiroshi Ohta, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  Journal of medical ultrasonics (2001) 41 (1) 11 - 21 2014/01 [Refereed][Not invited]
   

  PURPOSE: The aim of this study was to clarify whether ultrasound image-guided cisplatin delivery with an intratumor microbubble injection enhances the antitumor effect in a xenograft mouse model. METHODS: Canine thyroid adenocarcinoma cells were used for all experiments. Before in vivo experiments, the cisplatin and microbubble concentration and ultrasound exposure time were optimized in vitro. For in vivo experiments, cells were implanted into the back of nude mice. Observed by a diagnostic ultrasound machine, a mixture of cisplatin and ultrasound contrast agent, Sonazoid, microbubbles was injected directly into tumors. The amount of injected cisplatin and microbubbles was 1 μg/tumor and 1.2 × 10(7) microbubbles/tumor, respectively, with a total injected volume of 20 μl. Using the same diagnostic machine, tumors were exposed to ultrasound for 15 s. The treatment was repeated four times. RESULTS: The combination of cisplatin, microbubbles, and ultrasound significantly delayed tumor growth as compared with no treatment (after 18 days, 157 ± 55 vs. 398 ± 49 mm(3), P = 0.049). Neither cisplatin alone nor the combination of cisplatin and ultrasound delayed tumor growth. The treatment did not decrease the body weight of mice. CONCLUSION: Ultrasound image-guided anticancer drug delivery may enhance the antitumor effects of drugs without obvious side effects.
• Prognostic Value of Left Atrial Function in Dogs with Chronic Mitral Valvular Heart Disease

  Nakamura K, Osuga T, Morishita K, Suzuki S, Morita T, Yokoyama N, Ohta H, Yamasaki M, Takiguchi M

  Journal of Veterinary Internal Medicine Wiley-Blackwell 28 (6) 1746 - 1752 0891-6640 2014 [Refereed][Not invited]
   

  Background: A strong correlation between left atrial (LA) dysfunction and the severity of cardiac disease has been described in human patients with various cardiac diseases. The role of LA dysfunction in dogs with chronic mitral valvular heart disease (CMVHD) has not been addressed. Objectives: To investigate the correlation between LA function and the prognosis of dogs with CMVHD. Animals: Thirty-eight client-owned dogs with CMVHD. Methods: Prospective clinical cohort study. Dogs were divided into 2 groups (survivors and nonsurvivors) based on the onset of cardiac-related death within 1year. Physical examination and echocardiographic variables were compared between the groups. For the assessment of the comparative accuracy in identifying patients with cardiac-related death, receiver operating characteristic (ROC) curves and multivariate logistic analysis were used. Results: The highest accuracy was obtained for the LA active fractional area change (LA-FAC(act)), with an area under the ROC curve (AUC) of 0.95, followed by the left atrial to aortic root ratio (LA/Ao), with an AUC of 0.94; peak early diastolic mitral inflow velocity (E), with an AUC of 0.85; and LA total fractional area change (LA-FAC(total)), with an AUC of 0.85. In the multivariate logistic regression analysis, LA-FAC(act) emerged as the only independent correlate of cardiac-related death within 1year (odds ratio=1.401, P=.002). Conclusions and Clinical Importance: Regarding both the size and function, the LA has a strong correlation with the prognosis of dogs with CMVHD. The most significant independent predictor of mortality in this study was LA-FAC(act).
• Qualitative and Quantitative Contrast Enhanced Ultrasonography of the Pancreas Using Bolus Injection and Continuous Infusion Methods in Normal Dogs

  Sue Yee Lim, Kensuke Nakamura, Keitaro Morishita, Noboru Sasaki, Masahiro Murakami, Tatsuyuki Osuga, Hiroshi Ohta, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  JOURNAL OF VETERINARY MEDICAL SCIENCE 75 (12) 1601 - 1607 0916-7250 2013/12 [Refereed][Not invited]
   

  Quantitative contrast enhanced ultrasound is a major breakthrough for ultrasound imaging in recent years. However, contrast enhancement of the pancreas is brief with bolus injection. To assess if continuous infusion of Sonazoid (R) can prolong the duration of pancreatic enhancement over bolus injections, eight adult dogs received bolus injection and continuous infusion of Sonazoid (R) on separate days. Contrast enhanced ultrasound of the pancreatic parenchyma and proximal descending duodenum was performed, and time intensity curves reflecting tissue perfusions were generated. Perfusion parameters- time to initial upslope, peak time, time to wash-out and peak intensity were calculated and evaluated. Fast wash-in to intense peak, followed by rapid wash-out was observed for time intensity curves of bolus injection. With continuous infusion, contrast wash-in to peak intensity was gradual, followed by long plateau and slow wash-out. Median contrast enhancement durations of the pancreas and duodenum were significantly prolonged by continuous infusion from 11 sec (range, 10 to 23 sec) and 16 sec (range, 3 to 43 sec) at bolus injection to 205 sec (range, 170 to 264 sec, P<0.01) and 193 sec (range, 169 to 216 sec, P<0.05), respectively. Median peak intensity of the pancreas was 100.9 MPV (range, 80.2 to 124.3 MPV) at bolus injection and 77.6 MPV (range, 58.2 to 99.5 MPV, P<0.05) at continuous infusion. Prolonged continuous imaging is afforded by continuous infusion of contrast agent. Peak intensity of the pancreas was slightly diminished in continuous infusion, but offered adequate imaging subjectively.
• LC-MS/MS analysis of canine lipoproteins fractionated using the ultracentrifugation-precipitation method.

  Asuka Suto, Masahiro Yamasaki, Yukari Takasaki, Yasuyuki Fujita, Riichiro Abe, Hiroshi Shimizu, Hiroshi Ohta, Mitsuyoshi Takiguchi

  The Journal of veterinary medical science 75 (11) 1471 - 7 0916-7250 2013/11 [Refereed][Not invited]
   

  Due to the lack of a gold standard method in canine lipoprotein analysis, it is unclear whether canine high-density lipoprotein (HDL) and low-density lipoprotein (LDL) can be accurately evaluated by the lipoprotein analysis methods used for dogs. This study investigated whether the ultracentrifugation-precipitation (U-P) method was suitable as a gold standard method for analyzing canine lipoprotein. First, the U-P method was compared with a gel permeation high-performance liquid chromatography system (GP-HPLC). The concentrations of canine HDL cholesterol (HDL-C) and LDL cholesterol (LDL-C) determined by the U-P method correlated closely with those determined by GP-HPLC. However, the canine HDL-C concentration determined by the U-P method was lower than that determined by GP-HPLC, and the canine LDL-C concentration determined by the U-P method was higher than that determined by GP-HPLC. This study showed that some canine HDL could be precipitated with heparin manganese chloride solution. Second, the HDL and LDL fractions separated by the U-P method were analyzed by LC-MS/MS. The HDL fraction was found to contain only apolipoprotein A-I, which is an apolipoprotein of HDL, whereas the LDL fraction contained both apolipoprotein A-I and apolipoprotein B-100, which is an apolipoprotein of LDL. This data showed that a certain lipoprotein that includes apolipoprotein A-I might precipitate with canine LDL when using heparin manganese chloride solution. These results indicated that the U-P method is not currently a gold standard method for analyzing canine lipoproteins.
• Markedly increased expression of interleukin-8 in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds

  Yu Tamura, Hiroshi Ohta, Shidow Torisu, Masashi Yuki, Nozomu Yokoyama, Masahiro Murakami, Sue Yee Lim, Tatsuyuki Osuga, Keitaro Morishita, Kensuke Nakamura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 156 (1-2) 32 - 42 0165-2427 2013/11 [Refereed][Not invited]
   

  Inflammatory colorectal polyps (ICRPs) in miniature dachshunds were recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and/or space-occupying large polyps in the colorectal area and are thought to be a novel form of inflammatory bowel disease (IBD). To explore key mediators in the pathogenesis of ICRPs, we analyzed several pro-inflammatory cytokine (IL-1 beta, IL-6, TNF-alpha, IL-8, IL-12p35, IL-12/23p40, and IL-23p19) mRNA expressions in colorectal polyps in ICRP dogs by quantitative PCR. Among these cytokines, IL-8 mRNA expression was markedly up-regulated in large polyps. To examine IL-8 protein expression, we analyzed IL-8 protein level and its location in colorectal mucosal specimens of ICRP dogs by ELISA and immunofluorescence microscopy. IL-8 protein was significantly increased in large polyps and serum in dogs with ICRPs compared to controls. By immunofluorescence microscopy, IL-8 was only localized in macrophages, but not in mucosal epithelial cells or neutrophils. IL-8-positive macrophages were significantly increased in large polyps compared to controls. These results suggest that IL-8 is produced mainly by macrophages and may induce neutrophil infiltration in the colorectal area of ICRP dogs. (C) 2013 Elsevier B.V. All rights reserved.
• Expression of CD4(+) T cell cytokine genes in the colorectal mucosa of inflammatory colorectal polyps in miniature dachshunds

  Hiroshi Ohta, Kanae Takada, Shidow Torisu, Masashi Yuki, Yu Tamura, Nozomu Yokoyama, Tatsuyuki Osuga, Sue Yee Lim, Masahiro Murakami, Noboru Sasaki, Kensuke Nakamura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 155 (4) 259 - 263 0165-2427 2013/10 [Refereed][Not invited]
   

  Inflammatory colorectal polyps (ICRPs) in miniature dachshunds are recently recognized as a major cause of large bowel diarrhea in this dog breed in Japan. ICRPs are characterized by the formation of multiple small polyps and a space-occupying large polyp in the colorectal area, and are thought to be a novel form of inflammatory bowel disease (IBD). In humans, specific cytokine patterns attributed to T helper (Th)1, Th17 and regulatory T cells have important roles in the pathogenesis of IBD. Thus, the aim of the present study was to assess the gene expression of cytokines of T cell subsets in the colorectal mucosa from dogs with ICRPs. Colorectal mucosal specimens from 10 dogs with ICRPs and 14 control dogs were used in this study. Interferon (IFN)-gamma, interleukin (IL)-4, IL-17A and IL-10 mRNA expression was assessed using quantitative real-time PCR. IL-17A mRNA expression was significantly increased in large polyps compared to small polyps and controls. IFN-gamma and IL-10 mRNA expression in large polyps were significantly higher than in controls. There was no significant difference in IL-4 mRNA expression among the three groups. IL-17A is thought to play important roles in the pathogenesis of ICRPs. IL-10 up-regulation could oppose the proinflammatory function of IL-17A. (C) 2013 Elsevier B.V. All rights reserved.
• Increased intratumoral fluorothymidine uptake levels following multikinase inhibitor sorafenib treatment in a human renal cell carcinoma xenograft model

  Masahiro Murakami, Songji Zhao, Yan Zhao, Wenwen Yu, Chowdhury Nusrat Fatema, Ken-Ichi Nishijima, Masahiro Yamasaki, Mitsuyoshi Takiguchi, Nagara Tamaki, Yuji Kuge

  Oncology Letters 6 (3) 667 - 672 1792-1074 2013/09 [Refereed][Not invited]
   

  An early identification of the tumor response to sorafenib treatment is indispensable for selecting optimal personalized treatment strategies. However, at present, no reliable predictors are clinically available. 18F-fluorothymidine (18F-FLT) positron emission tomography (PET) is used to assess tumor proliferation, since the FLT uptake level reflects thymidine kinase-1 (TK-1) activity. Thus, the present study determined whether FLT was able to evaluate the early tumor response to sorafenib treatment in a human renal cell carcinoma (RCC A498) xenograft in comparison with the tumor proliferation marker, Ki-67. Mice bearing A498 tumors were assigned to the control and sorafenib-treated groups and the tumor volume was measured every day. [Methyl-3H(N)]-3′-fluoro-3′-deoxythymidine (3H-FLT) was injected 2 h prior to the sacrifice of the mice on days three and seven following the treatment. 3H-FLT autoradiography (ARG) and Ki-67 immunohistochemistry (IHC) were performed using tumoral 3H-FLT uptake level diffusely increased following the treatment, while no significant changes were observed in Ki-67 IHC. The intratumoral 3H-FLT uptake levels significantly increased by 2.7- and 2.6-fold on days three and seven following the treatment, while the tumor volume and Ki-67 index did not significantly change. Thus, an increased FLT uptake level was demonstrated following the treatment, which may indicate the suppression of thymidylate synthase (TS) and the compensatory upregulation of TK-1 activity by sorafenib.
• Repeatability and reproducibility of measurements obtained via two-dimensional speckle tracking echocardiography of the left atrium and time-left atrial area curve analysis in healthy dogs

  Tatsuyuki Osuga, Kensuke Nakamura, Sue Yee Lim, Yu Tamura, Wickramasekara Rajapakshage Bandula Kumara, Masahiro Murakami, Noboru Sasaki, Keitaro Morishita, Hiroshi Ohta, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  AMERICAN JOURNAL OF VETERINARY RESEARCH 74 (6) 864 - 869 0002-9645 2013/06 [Refereed][Not invited]
   

  Objective-To evaluate left atrial phasic function in healthy dogs by means of 2-D speckle tracking echocardiography with time-left atrial area curve analysis and to assess repeatability and reproducibility of obtained measurements. Animals-6 healthy Beagles. Procedures-Each dog underwent echocardiography twice on different days (3 nonconsecutive examinations/d). Images were analyzed with offline software; area of the left atrium was automatically calculated in each frame throughout the cardiac cycle to derive time-left atrial area curves. Variables used to assess left atrial phasic function (total, passive, and active emptying area and emptying fractions and mean active and total emptying rates) were calculated. Agreement between variables measured via speckle tracking echocardiography and a manual tracing method was assessed with modified Bland-Altman analysis. Withinday and between-day coefficients of variation were determined. Results-Mean +/- SD total, passive, and active emptying fractions of the left atrium were 49.8 +/- 3.5%, 27.7 +/- 4.0%, and 30.5 +/- 4.3%, respectively. Mean +/- SD total and active emptying rates were 16.0 +/- 2.5 cm(2)/s and 25.1 +/- 4.9 cm(2)/s, respectively. Within-day and between-day coefficients of variation were < 20% (range, 0.41% to 16.4%) for all variables except mean active emptying rate (between-day coefficient of variation, 29.2%). Agreement between variables measured via speckle tracking echocardiography and the manual tracing method was good, and differences between methods were nonsignificant. Conclusions and Clinical Relevance-Evaluation of left atrial phasic function via speckle tracking echocardiography was feasible; repeatability and reproducibility of measurements were adequate in healthy dogs. Studies are needed to determine clinical applicability in canine patients. (Am J Vet Res 2013;74:864-869)
• Evaluation of changes in the tumor microenvironment after sorafenib therapy by sequential histology and F-18-fluoromisonidazole hypoxia imaging in renal cell carcinoma

  Masahiro Murakami, Songji Zhao, Yan Zhao, Nusrat Fatema Chowdhury, Wenwen Yu, Ken-Ichi Nishijima, Mitsuyoshi Takiguchi, Nagara Tamaki, Yuji Kuge

  INTERNATIONAL JOURNAL OF ONCOLOGY 41 (5) 1593 - 1600 1019-6439 2012/11 [Refereed][Not invited]
   

  The mechanistic dissociation of 'tumor starvation' versus 'vascular normalization' following anti-angiogenic therapy is a subject of intense controversy in the field of experimental research. In addition, accurately evaluating changes of the tumor microenvironment after anti-angiogenic therapy is important for optimizing treatment strategy. Sorafenib has considerable anti-angiogenic effects that lead to tumor starvation and induce tumor hypoxia in the highly vascularized renal cell carcinoma (RCC) xenografts. F-18-fluoromisonidazole (F-18-FMISO) is a proven hypoxia imaging probe. Thus, to clarify early changes in the tumor microenvironment following anti-angiogenic therapy and whether F-18-FMISO imaging can detect those changes, we evaluated early changes in the tumor microenvironment after sorafenib treatment in an RCC xenograft by sequential histological analysis and F-18-FMISO autoradiography (ARG). A human RCC xenograft (A498) was established in nude mice, for histological studies and ARG, and further assigned to the control and sorafenib-treated groups (80 mg/kg, per os). Mice were sacrificed on Days 1, 2, 3 and 7 in the histological study, and on Days 3 and 7 in ARG after sorafenib treatment. Tumor volume was measured every day. F-18-FMISO and pimonidazole were injected intravenously 4 and 2 h before sacrifice, respectively. Tumor sections were stained with hematoxylin and eosin and immunohistochemically with pimonidazole and CD31. Intratumoral F-18-FMISO distribution was quantified in ARG. Tumor volume did not significantly change on Day 7 after sorafenib treatment. In the histological study, hypoxic fraction significantly increased on Day 2, mean vessel density significantly decreased on Day 1 and necrosis area significantly increased on Day 2 after sorafenib treatment. Intratumoral F-18-FMISO distribution significantly increased on Days 3 (10.2-fold, p<0.01) and 7 (4.1-fold, p<0.01) after sorafenib treatment. The sequential histological evaluation of the tumor microenvironment clarified tumor starvation in A498 xenografts treated with sorafenib. F-18-FMISO hypoxia imaging confirmed the tumor starvation. F-18-FMISO PET may contribute to determine an optimum treatment protocol after anti-angiogenic therapy.
• Link between inflammation and aquaporin-5 distribution in submandibular gland in sjögren's syndrome?

  M. S. Soyfoo, A. Konno, N. Bolaky, J. S. Oak, D. Fruman, C. Nicaise, M. Takiguchi, C. Delporte

  Oral Diseases 18 (6) 568 - 574 1354-523X 2012/09 [Refereed][Not invited]
   

  Oral Diseases (2012) 18, 568-574 Objective: To determine whether a link exists between inflammation and aquaporin-5 distribution in submandibular glands from three animal models for Sjögren's syndrome: IQI/JIC, r1ΔT/r2n and non-obese diabetic mice. Methods: Mice of different ages were used. Inflammatory infiltrates were quantified using the focus score. Acinar aquaporin-5 subcellular distribution was determined by immunohistochemistry and quantified using labelling indices. Results: Minor inflammatory infiltrates were present in r1f/r2n mice. Massive inflammatory infiltrates and acinar destruction were observed in 24-week-old non-obese diabetic mice, 10-and 13-month-old IQI/JIC mice and some r1ΔT/r2n mice. Aquaporin-5 immunoreactivity was primarily apical in submandibular glands from 8- and 24-week-old Balb/C mice, 8-week-old non-obese diabetic mice, 2-, 4- and 7-month-old IQI/JIC mice and r1f/r2n mice. In contrast, decreased apical aquaporin-5 labelling index with concomitant increased apical-basolateral, apical-cytoplasmic and/or apical-basolateral-cytoplasmic aquaporin-5 labelling indices was observed in 24-week-old non-obese diabetic, 10- and 13-month-old IQI/JIC and r1ΔT/r2n mice with a focus score≥1. Conclusions: Altered aquaporin-5 distribution in submandibular acinar cells from IQI/JIC, non-obese diabetic and r1ΔT/r2n mice with a focus score≥1 appears to be concomitant to the presence of inflammatory infiltrates and acinar destruction. © 2012 John Wiley & Sons A/S.
• Involvement of Mitochondrial Genes of Babesia gibsoni in Resistance to Diminazene Aceturate

  Bandula Kumara Wickramasekara Rajapakshage, Masahiro Yamasaki, Shiang-Jyi Hwang, Noboru Sasaki, Masahiro Murakami, Yu Tamura, Sue Yee Lim, Kensuke Nakamura, Hiroshi Ohta, Mitsuyoshi Takiguchi

  JOURNAL OF VETERINARY MEDICAL SCIENCE 74 (9) 1139 - 1148 0916-7250 2012/09 [Refereed][Not invited]
   

  The stability of the characteristics of the diminazene aceturate (DA)-resistant B. gibsoni isolate was initially determined in vitro. Part of the DA-resistant B. gibsoni isolate was cultured without DA for 4 weeks, and then newly exposed to 200 ng/m/ DA. As a result, this isolate could proliferate the same as the DA-resistant isolate, indicating that the characteristic of DA resistance was stable in the DA-resistant isolate. Additionally, the level of parasitemia in the DA-resistant isolate was comparatively lower than in the wild-type, suggesting that the proliferation potential of the DA-resistant isolate would be lower than that of the wild-type. Subsequently, to investigate the involvement of mitochondrial DNA (mtDNA) in DA resistance in B. gibsoni, the nucleotide sequences and deduced amino acid sequences of mitochondrial genes such as COXI, COXIII, and CYTb genes of the DA-resistant isolate, were compared with those of the wild-type. As a result, these three genes were not altered in the DA-resistant B. gibsoni isolate. Moreover, the transcription levels of COXI,COXIII, and CYTb genes were observed by semi-quantitative RT-PCR. As a result, the gene transcription of those genes in the DA-resistant isolate was not significantly altered. These results indicated that DA did not affect mtDNA directly in DA-resistant B. gibsoni. Thus, it is suggested that mtDNA should not be deeply involved in DA resistance in B. gibsoni.
• Bilateral Segmental Aplasia with Unilateral Uterine Horn Torsion in a Pomeranian Bitch

  Kensuke Nakamura, Masahiro Yamasaki, Tomohiro Osaki, Hiroshi Ohta, Noboru Sasaki, Keisuke Aoshima, Takashi Kimura, Mitsuyoshi Takiguchi

  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION 48 (5) 327 - 330 0587-2871 2012/09 [Refereed][Not invited]
   

  Bilateral segmental aplasia of the uterine horns with unilateral pyometra and uterine horn torsion were diagnosed in a Pomeranian bitch that presented with chronic abdominal distension and an acute onset of anorexia and lethargy. Because radiographic and ultrasonographic findings revealed the presence of markedly enlarged bilateral uterine horns filled with fluid in the caudal abdomen, a tentative diagnosis of either pyometra or hydrometra with uterine horn torsion was made. Exploratory laparotomy showed bilateral, segmentally distended uterine horns with unilateral uterine horn torsion. Ovariohysterectomy was performed, and bilateral segmental aplasia of the uterine horns with the development of unilateral uterine horn torsion was diagnosed histopathologically. To the authors' knowledge, this is the first report of uterine horn torsion in conjunction with segmental aplasia of the uterine horn in a bitch. (J Am Anim Hosp Assoc 2012; 48:327-330. DOI 10.5326/JAAHA-MS-5771)
• Analysis of energy generation and glycolysis pathway in diminazene aceturate-resistant Babesia gibsoni isolate in vitro

  Bandula Kumara Wickramasekara Rajapakshage, Masahiro Yamasaki, Masahiro Murakami, Yu Tamura, Sue Yee Lim, Tatsuyuki Osuga, Noboru Sasaki, Kensuke Nakamura, Hiroshi Ohta, Mitsuyoshi Takiguchi

  JAPANESE JOURNAL OF VETERINARY RESEARCH 60 (2-3) 51 - 61 0047-1917 2012/08 [Refereed][Not invited]
   

  In our previous study, the level of parasitemia of the diminazene aceturate (DA)-resistant B. gibsoni isolate was continuously lower than that of the wild-type, indicating the possible alteration of energy metabolism in that isolate. Therefore, in the present study, the concentrations of ATP, glucose, lactate, and pyruvate, and the activities of lactate dehydrogenase and pyruvate kinase in the wild-type and DA-resistant isolate of B. gibsoni were measured and compared to investigate the amount of energy generation and the activity of the glycolysis pathway. As a result, the intracellular ATP and glucose concentrations in the DA-resistant B. gibsoni isolate were significantly higher than those in the wild-type. Meanwhile, the concentrations of lactate and pyruvate and the activities of lactate dehydrogenase and pyruvate kinase in the DA-resistant B. gibsoni isolate were not different from those in the wild-type. These results indicated that the DA-resistant B. gibsoni isolate contained a higher ATP concentration than the wild-type, but the activity of the glycolysis pathway was not altered in the DA-resistant B. gibsoni isolate. However, we could not determine the mechanism of the high energy production of the DA-resistant B. gibsoni isolate. Further studies on the energy metabolism of B. gibsoni are necessary to clarify the mechanism of the high energy production in the DA-resistant B. gibsoni isolate.
• Enhanced Cytotoxic Effect of Cisplatin Using Diagnostic Ultrasound and Microbubbles In Vitro

  Sasaki Noboru, Nakamura Kensuke, Murakami Masahiro, Lim Sue Yee, Ohta Hiroshi, Yamasaki Masahiro, Takiguchi Mitsuyoshi

  11TH INTERNATIONAL SYMPOSIUM ON THERAPEUTIC ULTRASOUND 1481 307 - 310 0094-243X 2012 [Refereed][Not invited]
  
• ACTIVATION OF MICROBUBBLES BY SHORT-PULSED ULTRASOUND ENHANCES THE CYTOTOXIC EFFECT OF CIS-DIAMMINEDICHLOROPLATINUM (II) IN A CANINE THYROID ADENOCARCINOMA CELL LINE IN VITRO

  Noboru Sasaki, Nobuki Kudo, Kensuke Nakamura, Sue Yee Lim, Masahiro Murakami, W. R. Bandula Kumara, Yu Tamura, Hiroshi Ohta, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  ULTRASOUND IN MEDICINE AND BIOLOGY 38 (1) 109 - 118 0301-5629 2012/01 [Refereed][Not invited]
   

  Ultrasound targeted microbubble destruction has succeeded in delivering drugs and genes. This study was designed to explore characteristics of ultrasound targeted microbubble destruction using short-pulsed diagnostic ultrasound. Canine thyroid adenocarcinoma cells were exposed to short-pulsed diagnostic ultrasound in the presence of cis-diamminedichloroplatinum (II) (cisplatin) and ultrasound contrast agent Sonazoid (R) microbubbles. The cytotoxic effect of cisplatin was enhanced by short-pulsed diagnostic ultrasound and microbubbles. Incubation time with microbubbles influenced the cytotoxic effect of cisplatin. However, exposure duration did not affect the cytotoxic effect of cisplatin. Therefore, short-pulsed diagnostic ultrasound may activate microbubbles near cells and deliver cisplatin into cells. In addition, activation of microbubbles may be concluded in a short time. Our results suggest that short exposure duration could be potentially sufficient to induce efficient drug delivery by ultrasound targeted microbubble destruction using short-pulsed diagnostic ultrasound. (E-mail: mtaki@vetmed.hokudai.ac.jp) (C) 2012 World Federation for Ultrasound in Medicine & Biology.
• Effects and Mechanisms of Action of Polyene Macrolide Antibiotic Nystatin on Babesia gibsoni In Vitro

  Masahiro Yamasaki, Norihisa Tamura, Kensuke Nakamura, Noboru Sasaki, Masahiro Murakami, Wickramasekara Rajapakshage, Bandula Kumara, Yu Tamura, Sue Yee Lim, Hiroshi Ohta, Mitsuyoshi Takiguchi

  JOURNAL OF PARASITOLOGY 97 (6) 1190 - 1192 0022-3395 2011/12 [Refereed][Not invited]
   

  Nystatin is a membrane-active polyene macrolide antibiotic and a channel-forming ionophore. Nystatin exhibits in vitro activity against Babesia gibsoni infecting normal canine erythrocytes containing low potassium (LK) and high sodium concentrations, i.e., LK erythrocytes. The calculated IC(50) value of nystatin against B. gibsoni infecting LK erythrocytes was 31.96 mu g/ml. The anti-babesial activity of nystatin disappeared when B. gibsoni in LK erythrocytes were incubated in culture media containing high potassium concentrations (HK). Moreover, when the parasites were harbored in canine HK erythrocytes, which contained high potassium and low sodium concentrations as a result of high Na-K-ATPase activity, the in vitro anti-babesial activities of nystatin also disappeared, apparently due to protection by HK erythrocytes. This suggested that nystatin could show in vitro anti-babesial activity against B. gibsoni by its ionophorous activity, the same as other ionophores such as valinomycin. Subsequently, the effects of nystatin on the host cells were observed. Nystatin could not modify the intracellular concentrations of potassium, sodium, adenosine triphosphate, or glucose in either LK or HK erythrocytes, although it caused weak hemolysis in HK erythrocytes. In addition, nystatin did not affect the survival of canine peripheral polymorphonuclear leukocytes. In conclusion, nystatin destroyed B. gibsoni by ionophorous activity but did not affect either canine erythrocytes or leukocytes in vitro.
• Effects of sildenafil citrate on five dogs with Eisenmenger's syndrome

  K. Nakamura, M. Yamasaki, H. Ohta, N. Sasaki, M. Murakami, W. R. Bandula Kumara, M. Takiguchi

  JOURNAL OF SMALL ANIMAL PRACTICE 52 (11) 595 - 598 0022-4510 2011/11 [Refereed][Not invited]
   

  Objectives: To determine the effect of sildenafil for dogs with Eisenmeger's syndrome and secondary erythrocytosis.
• Pathological correlations between podocyte injuries and renal functions in canine and feline chronic kidney diseases.

  Ichii O, Yabuki A, Sasaki N, Otsuka S, Ohta H, Yamasaki M, Takiguchi M, Namiki Y, Hashimoto Y, Endoh D, Kon Y

  Histology and histopathology Universidad de Murcia 26 (10) 1243 - 1255 0213-3911 2011/10 [Refereed][Not invited]
   

  Podocytes cover the glomerulus and their adjacent foot processes form a principal barrier called the slit diaphragm. Podocyte dysfunctions, including podocyte loss and slit diaphragm disruptions, induce chronic kidney diseases (CKD). In this study, we analyzed the correlations between podocyte injuries and renal dysfunctions in domestic carnivores. Dogs and cats were divided into normal and CKD groups according to renal histopathology and plasma creatinine values. Immunostaining results showed that linear reactions of slit diaphragm molecules, e.g., nephrin, podocin, and ACTN4, were parallel to glomerular capillaries in all animals. However, in dogs, reactions of nephrin and ACTN4 were changed to a granular pattern in the CKD group, and their intensities significantly decreased with the number of podocytes in the glomerulus. Moreover, the expression of nephrin and ACTN4 negatively correlated with creatinine. Real-time PCR analysis showed that nephrin mRNA expression in the kidneys of CKD dogs was significantly lower than that in normal animals, and negatively correlated with creatinine. Although no significant correlation between renal dysfunction and podocyte injury was detected in cats, histoplanimetric scores of tubulointerstitial lesions in CKD cats were higher than those in both normal cats and diseased dogs. Furthermore, mRNAs of WT1 and SD molecules were detected in urine from CKD animals. In conclusion, podocyte injuries such as podocytopenia and decreased expression of nephrin and ACTN4 in the glomerulus were more strongly correlated with renal dysfunction in dogs than in cats. These findings suggest that the CKD pathogenesis, especially susceptibilities to podocyte injuries, differed between dogs and cats.
• Detection of anti-Babesia gibsoni heat shock protein 70 antibody and anti-canine heat shock protein 70 antibody in sera from Babesia gibsoni-infected dogs

  Masahiro Yamasaki, Mikiko Ishida, Kensuke Nakamura, Noboru Sasaki, Masahiro Murakami, Wickramasekara Rajapakshage Bandula Kumara, Yu Tamura, Sue Yee Lim, Hiroshi Ohta, Mitsuyoshi Takiguchi

  VETERINARY PARASITOLOGY 180 (3-4) 215 - 225 0304-4017 2011/08 [Refereed][Not invited]
   

  Antibodies that recognized either Babesia gibsoni or canine red blood cell (RBC) 70-kilodalton (kDa) protein were detected in serum from acutely and chronically B. gibsoni-infected. In those sera, antibodies that reacted with recombinant B. gibsoni and canine heat shock protein 70 (rBgHsp70 and rcHsp70) were detected; therefore, B. gibsoni and canine RBC 70-kDa proteins seemed to be BgHsp70 and cHsp70, respectively. In infected dogs, the amounts of these antibodies increased after infection. Interestingly, polyclonal antibody raised against rBgHsp70 in two rabbits reacted not only with rBgHsp70 but also with rcHsp70 and native cHsp70 from canine RBCs. Because BgHsp70 showed high homology with cHsp70 (70.8%), anti-rBgHsp70 antibody might cross-react with cHsp70. Additionally, the localizations of both BgHsp70 and cHsp70 were observed by indirect fluorescence assay. As a result, cHsp70 was not found on the membrane surface of erythrocytes, suggesting that erythrocytes would not be targets of anti-cHsp70 antibody. Meanwhile, only exoerythrocytic parasites were stained by anti-rBgHsp70 antibody. This result showed that BgHsp70 would be expressed on the surface of parasites during the exoerythrocytic stage. These results indicated that BgHsp70 was a highly immunogenic protein in canine B. gibsoni infection, and that exoerythrocytic parasites might be targets of anti-BgHsp70 antibody. (C) 2011 Elsevier B.V. All rights reserved.
• Expression and subcellular localization of apical junction proteins in canine duodenal and colonic mucosa

  Hiroshi Ohta, Tomoki Yamaguchi, B. K. Wickramasekara Rajapakshage, Masahiro Murakami, Noboru Sasaki, Kensuke Nakamura, Shiang-Jyi Hwang, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  AMERICAN JOURNAL OF VETERINARY RESEARCH 72 (8) 1046 - 1051 0002-9645 2011/08 [Refereed][Not invited]
   

  Objective-To examine the expression and distribution of tight junction (TJ) and adherens junction (AM proteins in canine duodenal and colonic mucosa. Sample-Mucosa obtained from 4 healthy Beagles. Procedures-Biopsy specimens of the duodenum and colon were obtained via endoscopy from 4 healthy dogs. The expression patterns and subcelluar localization of claudin-1, -2, -3, -4, -5, -7, and -8; E-cadherin; and beta-catenin in the duodenum and colon were analyzed by use of immunoblotting and immunofluorescence microscopy. Results-In the duodenum, there was clear expression of claudin-3 and -5, E-cadherin, and beta-catenin proteins and weak expression of claudin-7 protein. In contrast, there was clear expression of claudin-2 and -3, E-cadherin, and beta-catenin proteins and weak expression of claudin-5 and -7 proteins in the colon, as determined by use of immunoblotting. As determined by the use of immunofluorescence microscopy, the duodenum and colon had staining for claudin-3 and -5, E-cadherin, and beta-catenin in the most apical region and staining for claudin-7 in the basolateral region. Staining for claudin-2 was also observed in the colon. Conclusions and Clinical Relevance-Information was provided about the expression patterns of TJ and AJ proteins in the duodenum and colon of clinically normal dogs. These results may provide valuable information for use in evaluating the importance of these TJ and AJ proteins in the pathogenesis of inflammatory bowel disease in dogs. (Am J Vet Res 2011;72:1046-1051)
• Altered expression of miR-146a correlates with the development of chronic renal inflammation.

  Ichii O, Yabuki A, Sasaki N, Otsuka S, Ohta H, Yamasaki M, Takiguchi M, Namiki Y, Hashimoto Y, Endoh D, Kon Y

  Kidney Int. 81 (3) 280 - 292 2011 [Refereed][Not invited]
  
• Babesia gibsoni: Detection in blood smears and formalin-fixed, paraffin-embedded tissues using deoxyribonucleic acid in situ hybridization analysis

  Masahiro Yamasaki, Yusuke Kobayashi, Kensuke Nakamura, Noboru Sasaki, Masahiro Murakami, Bandula Kumara Wickramasekara Rajapakshage, Hiroshi Ohta, Osamu Yamato, Yoshimitsu Maede, Mitsuyoshi Takiguchi

  EXPERIMENTAL PARASITOLOGY 127 (1) 119 - 126 0014-4894 2011/01 [Refereed][Not invited]
   

  In this study, we attempted to detect Babesia gibsoni in blood smears and formalin-fixed, paraffin-embedded tissues obtained from B. gibsoni-infected dogs using in situ hybridization. Using a digoxigenin-conjugated deoxyribonucleic acid (DNA) probe, both intraerythrocytic and exoerythrocytic parasites in the culture could be specifically stained in blood smears fixed with 4% phosphate-buffered paraformaldehyde. This indicated that genomic DNA extracted from the parasites could be detected using in situ hybridization. Moreover, the parasite could be specifically stained in paraffin-embedded spleen, lymph node, and kidney sections using in situ hybridization. Infected erythrocytes in blood vessels in the spleen and kidney, hemosiderin-laden macrophages in the spleen, and phagocytized erythrocytes, which seemed to be infected with the parasites, in lymph nodes were also specifically stained. This suggests that in situ hybridization can be utilized to investigate both the life cycle of B. gibsoni and the pathological condition of canine babesiosis. (C) 2010 Elsevier Inc. All rights reserved.
• Contrast-enhanced ultrasonography for characterization of focal splenic lesions in dogs.

  Nakamura K, Sasaki N, Murakami M, Bandula Kumara WR, Ohta H, Yamasaki M, Takagi S, Osaki T, Takiguchi M

  Journal of veterinary internal medicine / American College of Veterinary Internal Medicine 6 24 (6) 1290 - 1297 0891-6640 2010/11 [Refereed][Not invited]
   

  Nakamura K, Sasaki N, Murakami M, Bandula Kumara WR, Ohta H, Yamasaki M, Takagi S, Osaki T, Takiguchi M, <i>Journal of veterinary internal medicine / American College of Veterinary Internal Medicine</i>, 2010, vol. 24, no. 6, pp. 1290-1297
• Reduced transcript levels of the heat shock protein 70 gene in diminazene aceturate-resistant Babesia gibsoni variants under low concentrations of diminazene aceturate

  Shiang-Jyi Hwang, Masahiro Yamasaki, Kensuke Nakamura, Noboru Sasaki, Masahiro Murakami, Bandula Kumara Wickramasekara Rajapakshage, Hiroshi Ohta, Yoshimitsu Maede, Mitsuyoshi Takiguchi

  JAPANESE JOURNAL OF VETERINARY RESEARCH 58 (3-4) 155 - 164 0047-1917 2010/11 [Refereed][Not invited]
   

  In our previous report, we developed a diminazene aceturate (DA)-resistant Babesia gibsoni strain that was maintained in culture with 200 ng/ml DA. While developing this strain, we also obtained DA-resistant B. gibsoni variants, which were maintained in culture with DA from 1 to 175 ng/ml for more than 8 weeks. Because heat shock protein 70 (Hsp70) seems to play important roles in adaptation to a stress environment in protozoan parasites, in the present study, we examined the copy number of B. gibsoni Hsp70 (BgHsp70) transcripts of those DA-resistant variants using quantitative real-time reverse transcription-polymerase chain reaction. We found that when wild-type B. gibsoni was exposed to 1 ng/ml DA, the level of BgHsp70 transcripts was decreased at day 14. The copy number of BgHsp70 transcripts in the DA-resistant variant cultured with 1 ng/ml DA was significantly lower than in wild-type B. gibsoni, while those in DA-resistant variants increased with escalating doses of DA from 1 to 75 ng/ml, although they were lower than in wild-type B. gibsoni. However, those in DA-resistant variants cultured with > 125 ng/ml DA were almost the same as wild-type B. gibsoni. These results indicated that the transcript levels of the BgHsp70 gene might be reduced when the parasites are exposed to a low concentration of DA, and then might recover to the normal level after achieving resistance against DA. We expect that further study of the function of BgHsp70 will elucidate the mechanism of drug resistance against DA in B. gibsoni.
• Development and Characterization of a Strain of Babesia gibsoni Resistant to Diminazene Aceturate In Vitro

  Shiang-Jyi Hwang, Masahiro Yamasaki, Kensuke Nakamura, Noboru Sasaki, Masahiro Murakami, Bandula Kumara Wickramasekara Rajapakshage, Hiroshi Ohta, Yoshimitsu Maede, Mitsuyoshi Takiguchi

  JOURNAL OF VETERINARY MEDICAL SCIENCE 72 (6) 765 - 771 0916-7250 2010/06 [Refereed][Not invited]
   

  We attempted to develop a strain of Babesia gibsoni resistant to diminazene aceturate (DA), an anti-babesial drug, in vitro. Since the DA-sensitive B. gibsoni strain could survive and proliferate in culture medium containing 1 ng/ml DA, the concentration of DA was gradually increased from 1 to 200 ng/ml. The results showed that the parasites could survive and proliferate in the medium containing 200 ng/ml DA, which was much higher than the 50% inhibitory concentration (IC(50)) of DA for B. gibsoni. Subsequently, these parasites were removed from erythrocytes and exposed directly to 200 ng/ml DA. They were able to survive and invade fresh erythrocytes, though the DA-sensitive B. gibsoni strain did not survive. Based on these results, the parasites cultured within 200 ng/ml DA were determined to be a DA-resistant B. gibsoni strain. In addition, the IC(50) levels of clindamycin, doxycycline and pentamidine for the DA-resistant B. gibsoni strain were determined. The IC(50) levels of clindamycin, doxycycline and pentamidine for the DA-resistant strain were higher than those for the DA-sensitive strain. The IC(50) of pentamidine for the resistant strain was much greater than that for the DA-sensitive strain. These results indicated that the DA-resistant B. gibsoni strain could have resistance not only to DA, but also to other anti-babesial drugs. In conclusion, we successfully developed a DA-resistant B. gibsoni strain in vitro.
• Local overexpression of interleukin-1 family, member 6 relates to the development of tubulointerstitial lesions

  Osamu Ichii, Saori Otsuka, Nobuya Sasaki, Akira Yabuki, Hiroshi Ohta, Mitsuyoshi Takiguchi, Yoshiharu Hashimoto, Daiji Endoh, Yasuhiro Kon

  LABORATORY INVESTIGATION 90 (3) 459 - 475 0023-6837 2010/03 [Refereed][Not invited]
   

  Identification of factors that exacerbate a disease is important for the development of biomarkers. In this study, we discovered ectopic overexpression of interleukin-1 family, member-6 (IL-1F6) in several murine renal diseases. IL-1F6 participates in cytokine/chemokine production in the epithelium. In PCR array analysis for inflammatory mediators, Il1f6 showed the highest expression in the kidney of the B6. MRLc1 glomerulonephritis model. IL-1F6 was localized in the epithelium from the DCTs to CCDs, which showed tubular dilations or epithelial deciduations. Ultrastructual examination of the epithelial cells revealed that IL-1F6 was localized on the cytoplasmic ribosome, vesicles, and nucleus. In and around these tubules, we found infiltrations of CD3-positive T-cells and nestin- or alpha-smooth-muscle actin-positive mesenchymal cells. Expression of the IL-1F6 protein and Il1f6 mRNA in the kidney was increased by the development of TILs in the B6. MRLc1 model and in lupus (BXSB, NZB/WF1, and MRL/lpr), nephrotic syndrome (ICGN), and streptozotocin-induced diabetic models. IL-1F6 was also detected in the epithelia having squamous or deciduous contours in other organs such as the skin, esophagus, thymus, or uterus. In vitro analysis using M-1 cells from the murine collecting duct revealed that Il1f6 mRNA induction was related to the upregulation of IL-6, TGF-beta receptor-1, and mesenchymal markers and to the downregulation of epithelial markers and changes in the squamous cells of the epithelium. Interestingly, urine Il1f6 mRNA expression was detected earlier than renal dysfunctions in these mouse models. Ectopic overexpression of IL-1F6 in kidneys is associated with TILs and especially with cell infiltrations and changes in epithelial morphology. We propose that local overexpression of IL-1F6 is related to the development of TILs.
• CONTRAST-ENHANCED ULTRASONOGRAPHY FOR CHARACTERIZATION OF CANINE FOCAL LIVER LESIONS

  Kensuke Nakamura, Satoshi Takagi, Noboru Sasaki, Wickramasekara Rajapakshage Bandula Kumara, Masahiro Murakami, Hiroshi Ohta, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  VETERINARY RADIOLOGY & ULTRASOUND 51 (1) 79 - 85 1058-8183 2010/01 [Refereed][Not invited]
   

  In six normal beagles and 27 dogs with spontaneous focal or multifocal liver lesions, contrast-enhanced ultrasonography using Sonazoid (R) was performed. Sonazoid (R) is a newly developed second-generation contrast agent with the ability to be used for real-time contrast imaging along with parenchymal imaging. An appropriate protocol for the evaluation of all three phases (arterial, portal, and parenchymal) was established based on the results for normal beagles. By evaluation of the echogenicity of hepatic nodules during the arterial and parenchymal phases it was possible to differentiate malignant tumors from benign nodules with very high accuracy. In 15 of 16 dogs diagnosed as malignant tumors, nodules were clearly hypoechoic to the surrounding normal liver during the parenchymal phase. Additionally, malignant tumors had different echogenicity compared with the surrounding normal liver during the arterial phase in 14 of 15 dogs. In the portal phase, there were no characteristic findings. Contrast-enhanced ultrasonography with Sonazoid (R) appears to improve the characterization of canine focal and multifocal hepatic lesions.
• EFFECTS AND MECHANISMS OF ACTION OF IONOPHOROUS ANTIBIOTICS VALINOMYCIN AND SALINOMYCIN-NA ON BABESIA GIBSONI IN VITRO

  Masahiro Yamasaki, Kensuke Nakamura, Norihisa Tamura, Shiang-Jyi Hwang, Muneyoshi Yoshikawa, Noboru Sasaki, Hiroshi Ohta, Osamu Yamato, Yoshimitsu Maede, Mitsuyoshi Takiguchi

  JOURNAL OF PARASITOLOGY 95 (6) 1532 - 1538 0022-3395 2009/12 [Refereed][Not invited]
   

  Valinomycin and salinomycin-Na, 2 ionophorous antibiotics, exhibited in vitro antibabesial activities against Babesia gibsoni that infected normal canine erythrocytes containing low potassium (LK) and high sodium concentrations, i.e., LK erythrocytes, which completely lack Na,K-ATPase activity. The level of parasitemia of B. gibsoni was significantly decreased when the parasites were incubated in culture medium containing either 10(-1) ng/ml valinomycin or 10(2) ng/ml salinomycin-Na for 24 hr. Four-hour incubation in the culture medium containing 5 mu g/ml salinomycin-Na led to the destruction of most parasites. In contrast, when the parasites infected canine erythrocytes containing high potassium (HK) and low sodium concentrations, i.e., HK erythrocytes, the in vitro antibabesial activities of both ionophorous antibiotics seemed to be weakened, apparently due to the protection by the host cells. Therefore, differential influences of ionophorous antibiotics on LK and HK erythrocytes were observed. In LK erythrocytes, the intracellular concentrations of potassium, sodium, and adenosine triphosphate (ATP) were not modified, and hemolysis was not observed after incubation in the medium containing each ionophorous antibiotic. These results suggested that these ionophorous antibiotics did not affect cells without Na,K-ATPase, and directly affected B. gibsoni. In HK erythrocytes, the ionophorous antibiotics increased the intracellular sodium concentration, and decreased the intracellular potassium and ATP concentrations, causing obvious hemolysis. Additionally, the decrease of the intracellular ATP concentration and the hemolysis in HK erythrocytes caused by valinomycin disappeared when the activity of Na,K-ATPase was inhibited by ouabain. These results indicate that modification of the intracellular cation concentrations by the ionophorous antibiotics led to the activation of Na,K-ATPase and increased consumption of intracellular ATP, and that the depletion of intracellular ATP resulted in hemolysis in HK erythrocytes. Moreover, the antibabesial activity of valinomycin disappeared when B. gibsoni in LK erythrocytes were incubated in culture media containing high potassium concentrations. This showed that the intracellular cation concentration in the parasites was not modified in those media and would remain the same.
• Primary Splenic Torsion in a Boston Terrier

  Hiroshi Ohta, Satoshi Takagi, Masahiro Murakami, Noboru Sasaki, Muneyoshi Yoshikawa, Kensuke Nakamura, Shiang-Jyi Hwang, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  JOURNAL OF VETERINARY MEDICAL SCIENCE 71 (11) 1533 - 1535 0916-7250 2009/11 [Refereed][Not invited]
   

  A 7-year-old female Boston terrier was referred to Hokkaido University Veterinary Teaching Hospital with a history of hemoglobinuria and anemia for several days. Abdominal radiographs showed splenomegaly, and Ultrasonography revealed a hypoechoic splenic parenchyma with interspersed linear echoes consistent with the ultrasonographic appearance of splenic torsion. Ultrasonography and computed tomography (CT) indicated a C-shaped spleen. Exploratory laparotomy confirmed the diagnosis of splenic torsion. A splenectomy was performed, and the dog recovered well Without complications. This is the first report of splenic torsion in Boston terriers, and the usefulness of ultrasonographic and CT findings of the splenic torsion was also confirmed.
• QUANTITATIVE CONTRAST-ENHANCED ULTRASONOGRAPHY OF CANINE SPLEEN

  Kensuke Nakamura, Noboru Sasaki, Muneyoshi Yoshikawa, Hiroshi Ohta, Shiang-Jyi Hwang, Takahiro Mimura, Masahiro Yamasaki, Mitsuyoshi Takiguchi

  VETERINARY RADIOLOGY & ULTRASOUND 50 (1) 104 - 108 1058-8183 2009/01 [Refereed][Not invited]
   

  Contrast-enhanced ultrasonography has an important role in the detection of tumors in humans. The second-generation contrast agent Sonazoid has the ability of real-time contrast imaging along with parenchymal imaging. The purposes of this study were to determine the effect and duration of Sonazoid on the changes in gray-scale enhancement of canine spleen and to establish an appropriate protocol for contrast-enhanced ultrasonography of canine spleen. Six healthy beagles were injected with an intravenous bolus of Sonazoid. In the spleen parenchyma, the enhancement was maintained up to 30 min after injection. Moreover, for 5-22 s after injection, gray-scale enhancement of splenic arteries afforded arterial imaging. Perfusion of the kidney may be investigated from 3.6 s to 3.5 min after injection of Sonazoid. These results suggest that Sonazoid is applicable to canine spleen parenchymal imaging and that the optimal time for the parenchymal imaging is 7-30 min after injection. The findings of this quantitative study should prove useful in the evaluation of diffuse or focal splenic and renal diseases in dogs.
• Flow cytometry to evaluate the level of Babesia gibsoni parasitemia in vivo and in vitro by using the fluorescent nucleic acid stain SYTO16

  Masahiro Yamasaki, Shiang-Jyi Hwang, Hiroshi Ohta, Osamu Yamato, Yoshimitsu Maede, Mitsuyoshi Takiguchi

  JAPANESE JOURNAL OF VETERINARY RESEARCH 55 (4) 129 - 136 0047-1917 2008/02 [Refereed][Not invited]
   

  In the present study, we employed flow cytometry to evaluate the level of parasitemia of Babesia gibsoni infecting canine erythrocytes in vivo and in vitro by using fluorescent nucleic acid staining. Peripheral blood samples from a R gibsoni-infected dog and cultured B. gibsoni parasitizing in canine erythrocytes were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16. In this study, we utilized normal canine erythrocytes (LK erythrocytes) and canine erythrocytes containing high concentrations of potassium, reduced glutathione, and some free amino acids (HK erythrocytes) as host cells for culture. Parasitized cells in vivo were discriminated completely from unparasitized cells and a correlation (r = 0.998) between the percentage of SYTO16-positive cells and parasitemia in vivo was observed. On the other hand, erythrocytes in vitro could not be divided clearly into parasitized and unparasitized cells. However, when LK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost the same as, and was well correlated (r = 0.932) with, the level of parasitemia. When HK erythrocytes were used as host cells, the percentage of SYTO16-positive cells was almost half of, but was correlated (r = 0.982) with, the level of parasitemia. Therefore, we attempted to observe the changes in the percentage of parasitized cells after treatment with antiprotozoal drug or mitochondria. inhibitors by using flow cytometry. The changes in the percentage of SYTO16-positive cells corresponded well with the changes of the level of parasitemia when the parasites in HK erythrocytes were cultured with each compound. The present results suggest that flow cytometric detection using SYTO16 is a rapid and reliable method for monitoring parasitemia both in vivo and in vitro.
• Identification of a quantitative trait locus regulating B cell-dominant infiltration into autoimmune sialitis lesions of the IQI mouse model of primary Sjogren's syndrome

  Akihiro Konno, Mitsuyoshi Takiguchi, Kensuke Takada, Takeshi Usami, Kaoru Azumi, Hisayo Kubota, Mutsumi Inaba, Junzo Saegusa, Yasuhiro Kon

  IMMUNOGENETICS 59 (11) 853 - 859 0093-7711 2007/11 [Refereed][Not invited]
   

  Sjogren's syndrome (SS) is caused by an autoimmune sialodacryoadenitis, and up to 5% of patients with SS develop malignant B cell growth. The IQI mouse is a spontaneous model of primary SS in which B cells are the dominant cellular subpopulation among mononuclear infiltrates in sialitis lesions. Understanding the genetic control of aberrant B cell growth in IQI mice may help elucidate the genetic mechanisms involved in B-lineage hyperplasia leading to malignant transformation in human SS. B cell-dominant infiltration in the submandibular glands of 6-month-old IQI and C57BL/6 (B6) mice and their F1 and F2 progenies was quantified as B-lymphocytic sialitis score, and a genome-wide scan of 179 (IQI x B6) F2 females was performed to identify a quantitative trait locus (QTL) controlling this phenotype. A QTL significantly associated with variance in B-lymphocytic sialitis score was mapped to the D6Mit138 marker (position of 0.68cM) on proximal chromosome 6, with a logarithm of odds score of 4.3 (p=0.00005). This QTL, named autoimmune sialitis in IQI mice, associated locus 1 (Asq1), colocalized with Islet cell autoantigen 1 (Ica1), which encodes a target protein of the immune processes that define the pathogenesis of primary SS in humans and in the nonobese diabetic mouse model.
• Restricted localization of claudin-16 at the tight junction in the thick ascending limb of Henle's loop together with claudins 3, 4, and 10 in bovine nephrons

  H Ohta, H Adachi, M Takiguchi, M Inaba

  JOURNAL OF VETERINARY MEDICAL SCIENCE 68 (5) 453 - 463 0916-7250 2006/05 [Refereed][Not invited]
   

  Claudin-16 is one of the tight junction protein claudins and has been shown to contribute to reabsorption of divalent cations in the human kidney. In cattle, total deficiency of claudin-16 causes severe renal tubular dysplasia without aberrant metabolic changes of divalent cations, suggesting that bovine claudin-16 has some roles in renal tubule formation and paracellular transport that are somewhat different from those expected from the pathology of human disease. As the first step to clarify these roles, we examined the expression and distribution of claudin-16 and several other major claudin subtypes. claudins 1-4 and 10. in bovine renal tubular segments by immunofluorescence microscopy. Claudin-16 was exclusively distributed to the tight junction in the tubular segment positive for Tamm-Horsfall glycoprotein, the thick ascending limb (TAL) of Henle's loop. and was found colocalized with claudins 3. 4. and 10. This study also demonstrates that bovine kidneys possess segment-specific expression patterns for claudins 2-1 and 10 that are different from those reported for mice. Particularly, distribution of claudin-4 in the TAL and distal convoluted tubules was characteristic of bovine nephrons as were differences in the expression patterns of claudins 2 and 3. These findings demonstrate that the total lack of claudin-16 in the TAL segment is the sole cause of renal tubular dysplasia in cattle and suggest that the tight junctions in distinct tubular segments including the TAL have barrier functions in paracellular permeability that are different among animal species.
• Magnetic resonance imaging of alveolar echinococcosis experimentally induced in the rat lung

  T Asanuma, T Kawahara, O Inanami, M Nakao, K Nakaya, A Ito, M Takiguchi, A Hashimoto, M Kuwabara

  JOURNAL OF VETERINARY MEDICAL SCIENCE 68 (1) 15 - 20 0916-7250 2006/01 [Refereed][Not invited]
   

  Pulmonary alveolar echinococcosis (AE) caused by the metacestode of Echinococcus multilocularis is a lethal zoonosis and 14 is a lesion secondarily induced by hematogenous dissemination from hepatic AE lesions. In the present study, a hematogenous pulmonary AE model was experimentally induced in rats by the injection of echinococcal larval tissue homogenate to the tail vein, and then the pathological and diagnostic aspects of pulmonary AE were examined by magnetic resonance imaging (MRI). Histological primary, mature and degenerated AE lesions were observed 5, 18 and 50 weeks after injection, respectively. These lesions were discriminated as signal-void, hypointense and hyperintense regions in T1-weighted MRI (T1WI), respectively. The change in signal intensity in T1WI 1 might reflect the content of proteinaceous fluid as a result of AE cyst degeneration. Western blot analysis of sera with antibodies of two epitopes (Em18 and Em16) of E. multilocularis provided evidence for AE infection in the early stage. T1WI in combination with Western blot analysis could possibility become definitive and early signs of hematogenous pulmonary A-E infection.
• Different effects on the inflammatory lesions in the lacrimal and salivary glands after neonatal thymectomy in IQI/Jic mice, a model for Sjogren's syndrome

  K Takada, M Takiguchi, M Inaba

  JOURNAL OF VETERINARY MEDICAL SCIENCE 67 (9) 955 - 957 0916-7250 2005/09 [Refereed][Not invited]
   

  Thymectomy on day 3 after birth (D3Tx) is understood to eliminate CD4(+)CD25(+) regulatory T cells (T-reg) from the peripheral T cell repertoire in rodents, leading to the activation of autoreactive T cells. Herein, D3Tx was performed in IQI/Jic mice, a model for Sjogren's syndrome characterized by autoimmune infiltrations into the lacrimal and salivary glands. At the age of 16 weeks, very severe lesions were observed in lacrimal tissues from thymectornized mice, suggesting that T-reg preserve their immunoregulatory function in young IQI/Jic mice. In contrast, salivary lesions were comparable in the D3Tx and control groups, raising the possibility that either salivary-specific T-reg escaped elimination in thymectornized mice or spontaneous lesions in IQI/Jic mice developed independently of the tolerance through T-reg.
• Autoimmunity against a tissue kallikrein in IQI/Jic mice - A model for Sjogren's syndrome

  K Takada, M Takiguchi, A Konno, A Inaba

  JOURNAL OF BIOLOGICAL CHEMISTRY 280 (5) 3982 - 3988 0021-9258 2005/02 [Refereed][Not invited]
   

  We have recently characterized IQI/Jic mice as a model for Sjogren's syndrome (SS), a chronic autoimmune disease in humans. In SS, local lymphocytic infiltrations into salivary and lacrimal glands frequently develop to the involvement of systemic exocrine and nonexocrine organs, and the mechanism for progression of this disease remains obscure. Herein, we report identification of an autoantigen shared by various target organs in IQI/Jic mice. Polypeptides identified based on immunorecognition by autoantibodies in sera from IQI/ Jic mice affected with autoimmune disease (> 12 weeks of age) were tissue kallikrein (Klk)-1 and -13 and were cross-reactive to the autoantibodies. Interestingly, Klk-13, but not Klk-1, caused a proliferative response of splenic T cells from IQI/Jic mice from the age of 4 weeks onward. In addition, remarkably enhanced expression of Klk-13 was observed in the salivary glands of the mice in accordance with the development of inflammatory lesions. These results indicate that Klk-13 acts as an autoantigen and may increase T cells responsive to organs commonly expressing Klk-13, playing a pivotal role in the etiology of progression of disease in IQI/Jic mice. Our findings provide insights into the contributions of autoantigens shared by multiple organs in the progress of SS from an organ-specific to a systemic disorder.
• Diagnostic ultrasound of polypoid cystitis in dogs

  M Takiguchi, M Inaba

  JOURNAL OF VETERINARY MEDICAL SCIENCE 67 (1) 57 - 61 0916-7250 2005/01 [Refereed][Not invited]
   

  Polypoid cystitis is a rare disease of the urinary bladder in dogs characterized by chronic inflammation, epithelial proliferation, and development of a polypoid mass or masses without histopathologic evidence of neoplasia. The ultrasonographic appearances of eight dogs with polypoid cystitis are described. Ultrasonography confirmed the presence of a bladder mass or masses in all patients. Ultrasonographic findings are mucosal projections and a polypoid to pedunculated mass of variable size and shape. Although a polypoid mass tends to be located in the cranioventral bladder mucosa, the polyps also could arise in the craniodorsal bladder mucosa. Ultrasonographic images are well correlated with contrast radiographic studies and gross morphological appearance. Ultrasound is a non-invasive, very useful diagnostic tool for detecting bladder polyps, but histopathology is required for definitive diagnosis.
• Magnetic resonance imaging and immunoblot analyses in rats with experimentally induced cerebral alveolar echinococcosis

  T Asanuma, Y Matsumoto, M Takiguchi, O Inanami, M Nakao, K Nakaya, A Ito, A Hashimoto, M Kuwabara

  COMPARATIVE MEDICINE 53 (6) 649 - 656 1532-0820 2003/12 [Refereed][Not invited]
   

  The early stage of experimentally induced secondary cerebral alveolar echinococcosis (AE) in rats was investigated by use of magnetic resonance imaging (MRI) and immunoblot (western blot) analyses. Thirty-six female Wistar rats (6 to 8 weeks old) were injected intracranially with a 10% homogenate of echinococcal larval tissues in which the concentrations of microvesicles and protoscolices were estimated to be 3.8 and 1.5 x 10(4)/ml, respectively. To observe the fine structure of the rat brain, MRI was performed under a high magnetic field of 7.05 T. Histologic examination also was performed. The T2-weighted MR images revealed a hyperintense region in the cerebral cortex at two weeks after injection of the homogenate. At three weeks after injection, this region was found to have cysts on the basis of results of histologic examination. Signal-void regions corresponding to hyperplasia and the subsequent calcification of the cuticle layer at six and 13 weeks after injection, respectively, were observed in T2-weighted and proton density MR images. On the other hand, at nine weeks after injection, AE was discernible by use of western blot analysis of sera with antibodies of two epitopes (Em18 and Em16) of E. multilocularis. Using this secondary cerebral AE animal model, it was concluded that the MRI method was suitable for early detection of secondary cerebral AE.
• Presence of B7-2+ dendritic cells and expression of Th1 cytokines in the early development of sialodacryoadenitis in the IQI/Jic mouse model of primary Sjögren's syndrome

  Akihiro Konno, Kensuke Takada, Junzo Saegusa, Mitsuyoshi Takiguchi

  Autoimmunity 36 (4) 247 - 254 0891-6934 2003/06 [Refereed][Not invited]
   

  Subpopulations of infiltrating lymphocytes, professional antigen-presenting cells (APCs), and Th1/Th2 cytokines that could initiate an autoimmune sialodacryoadenitis were studied in the IQI/Jic mouse model of primary Sjogren's syndrome. Although lymphocytic infiltrations were first seen in submandibular glands (SMGs) of females and in lacrimal glands (LGs) of males at 8 weeks of age, clusters of MHC class II+, GD11c+, B7-2+ dendritic cells (DCs) were already localized in these tissues at 4 weeks. At 8 weeks, the infiltrating lymphocytes consisted of almost equal numbers of B cells and CD4+ T cells. In the inflammatory foci, MHC class II+, CD11c+, B7-2+ DCs formed network-like structures. Duct cells in the lesions showed immunoreactivities for MHC class II and ALCAM (a costimulatory adhesion molecule). IL-12 and IFN-γ transcripts were detected by RT-PCR in SMGs of females and in LGs of males at 8-12 weeks. These results suggest that the clustered DCs might play an important role in the initiation of the adenitis, and further suggest that the DCs and epithelial cells may participate in the activation of CD4+ T cells. It is also likely that Th1 cytokines mediate the functional interactions between the APCs and CD4+ T cells in the early lesions.
• Virologic and serologic identification of minute virus of canines (canine parvovirus type 1) from dogs in Japan

  M Mochizuki, M Hashimoto, T Hajima, M Takiguchi, A Hashimoto, Y Une, F Roerink, T Ohshima, CR Parrish, LE Carmichael

  JOURNAL OF CLINICAL MICROBIOLOGY 40 (11) 3993 - 3998 0095-1137 2002/11 [Refereed][Not invited]
   

  Minute virus of canines (MVC), also known as canine parvovirus type 1, was initially believed to be a nonpathogenic agent, since it was first isolated from canine fecal specimens in the late 1960s. However, subsequent pathological as well as epidemiological studies suggested that MVC is a pathogen of neonatal puppies and is widely distributed among domestic dogs in the United States. The virus also has been shown to cause fetal deaths. Nevertheless, the virus was not detected in dogs outside the United States until recently, presumably because of a lack of widespread availability of the only susceptible canine cell line, WRCC/3873D, used for MVC isolation. We examined 470 clinical specimens from 346 dogs by PCR and detected MVC-specific gene fragments from four diseased puppies (positive rate, 1.2%). Viruses were recovered from three PCR-positive rectal specimens by using WRCC/3873D and MDCK cells. The isolates possessed antigenic and genomic properties similar to those of the U.S. reference strain GA3 and were identified as MVC. In addition, seroepidemiological evidence that 5.0% of dogs possessed anti-MVC antibodies also indicated the presence of MVC infection among dogs in Japan. From this study and several recent European reports describing MVC field cases, it is evident that MVC is distributed among domestic dogs worldwide.
• Detection of Canine Herpesvirus DNA in the Ganglionic Neurons and the Lymph Node Lymphocytes of Latently Infected Dogs

  Masahiro Miyoshi, Yuki Ishii, Mitsuyoshi Takiguchi, Ayato Takada, Jun Yasuda, Akira Hashimoto, Katsunori Okazaki, Hiroshi Kida

  Journal of Veterinary Medical Science 61 (4) 375 - 379 1782-155X 1999/04 [Refereed][Not invited]
   

  To determine the site of latent infection of canine herpesvirus (CHV), tissues from dogs convalescent from acute infection with CHV were examined for the presence of viral genome DNA by the nested polymerase chain reaction. CHV DNA was detected in the trigeminal ganglia and the retropharyngeal lymph nodes. In situ hybridization study of the tissues revealed that CHV genome persisted in the nuclei of ganglionic neurons and lymphocytes.
• イヌヘルペスウイルスの神経節神経細胞及びリンパ節リンパ球における潜伏感染

  三好 正浩, 石井 由紀, 滝口 満喜, 高田 礼人, 安田 隼, 橋本 晃, 岡崎 克則, 喜田 宏

  The Journal of Veterinary Medical Science 61 (4) 375 - 379 0916-7250 1999/04 [Refereed][Not invited]
  

MISC

• 乳び胸の原因としてリンパ管腫症が疑われた犬の1例

  早坂 郁人, 新坊 弦也, 細谷 謙次, 三木 伸悟, 滝口 満喜  日本獣医画像診断学会抄録  73回-  np176  -np176  2023/12
• 犬の腎うっ血の評価におけるShear wave elastographyの有用性の検討

  塩原 希, 中村 健介, 横山 望, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  67-  (8)  316  -316  2023/08
• 拘束型心筋症に伴う左室内血栓により大動脈血栓塞栓に陥った猫の1例

  立花 圭, 中村 健介, 塩原 希, 川上 侑記, 滝口 満喜  北海道獣医師会雑誌  67-  (8)  318  -318  2023/08
• コンパニオンアニマルのPFAS汚染:イヌ・ネコの種差要因と汚染の地域差

  佐藤 愛佳, 佐藤 楓夏, 水川 葉月, 池中 良徳, 田上 瑠美, 横山 望, 滝口 満喜, 中津 賞, 野見山 桂  環境化学物質3学会合同大会要旨集  2回-  305  -306  2023/05
• 胃・小腸の拡張—特集 画像所見からみる診断のアプローチ

  横山 望, 滝口 満喜  伴侶動物画像診断 = Diagnostic imaging of companion animal  8-  (2)  6  -32  2023/04
• 造影超音波検査で副腎皮質の血流消失を確認した医原性副腎皮質機能低下症の犬の1例

  永田矩之, 前田貞俊, 青島圭佑, 中村健介, 滝口満喜  日本産業動物獣医学会(中部)・日本小動物獣医学会(中部)・日本獣医公衆衛生学会(中部)  2023-  2023
• 犬の発作性ジスキネジア23例の回顧的検討

  川上侑記, 笹岡一慶, 大田寛, 山根由久, 新坊弦也, 横山望, 佐々木東, 森下啓太郎, 中村健介, 滝口満喜  北海道獣医師会雑誌  67-  (8)  2023
• 犬尿路上皮癌におけるPDXモデルの樹立

  森下 大暉, 木之下 怜平, 青島 圭佑, 細谷 謙次, 山崎 淳平, 滝口 満喜  日本獣医学会学術集会講演要旨集  165回-  [B2P  -03]  2022/09
• 容量過負荷状態の犬における心エコー図法による左心房stiffnessの推定

  大菅 辰幸, 田村 昌大, 佐々木 東, 滝口 満喜  日本獣医学会学術集会講演要旨集  165回-  [HS1P  -05]  2022/09
• 2次診療動物病院に来院した犬の糞便真核生物叢解析

  横山 望, 川口 剛, 中尾 亮, 森下 啓太郎, 細谷 謙次, 永田 矩之, 笹岡 一慶, 新坊 弦也, 佐々木 東, 中村 健介, 滝口 満喜  日本獣医学会学術集会講演要旨集  165回-  [HS1P  -16]  2022/09
• 犬尿路上皮癌におけるPDXモデルの樹立

  森下 大暉, 木之下 怜平, 青島 圭佑, 細谷 謙次, 山崎 淳平, 滝口 満喜  日本獣医学会学術集会講演要旨集  165回-  [B2P  -03]  2022/09
• 低コレステロール血症は犬の膵外分泌不全を予測できるのか?

  横山 望, 大田 寛, 永田 矩之, 笹岡 一慶, 新坊 弦也, 佐々木 東, 森下 啓太郎, 中村 健介, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  284  -284  2022/08
• 洞停止を認めた咳嗽失神の犬の1例

  幸野 瑠美, 中村 健介, 笹岡 一慶, 塩原 希, 小幡 祐路, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  284  -284  2022/08
• 不完全型房室中隔欠損症の外科的整復時に両側房室弁の弁輪形成術を行った犬の1例

  川元 誠, 中村 健介, 森 拓也, 古越 真耶, 大橋 美里, 三木 悠矢, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  285  -285  2022/08
• 甲状腺機能低下症の治療により失神の消失を認めた犬の1例

  塩原 希, 中村 健介, 幸野 瑠美, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  285  -285  2022/08
• 臨床的に間質性肺疾患と診断された犬25頭の回顧的検討

  川上 侑記, 中村 健介, 新坊 弦也, 山根 由久, 大菅 辰幸, 竹内 恭介, 木之下 怜平, 細谷 謙次, 永田 矩之, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  286  -286  2022/08
• 犬尿中コルチゾール/クレアチニン比の基準値の確立とクッシング症候群の診断への応用

  永田 矩之, 澤村 飛龍, 五日市 大介, 森下 啓太郎, 細谷 謙次, 横山 望, 笹岡 一慶, 佐々木 東, 中村 健介, 池中 良徳, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  286  -286  2022/08
• 院内湿潤環境に対する注意喚起 北大附属動物病院における環境サーベイランス結果

  笹岡 一慶, 新田 優里香, 佐藤 豊孝, 堀内 基広, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  287  -287  2022/08
• 磁気共鳴スペクトロスコピーは犬の起源不明髄膜脳脊髄炎の予後指標となり得るか

  山口 敬央, 笹岡 一慶, 新坊 弦也, 山根 由久, 永田 矩之, 横山 望, 森下 啓太郎, 佐々木 東, 中村 健介, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  287  -287  2022/08
• 造影CT検査後に非心原性肺水腫を呈した褐色細胞腫を疑う犬の1例

  吉田 紫乃, 田村 純, 永田 矩之, 大山 紀彦, 新坊 弦也, 池中 良徳, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  289  -289  2022/08
• 麻酔中パルスオキシメーター測定異常から舌局所の灌流障害を疑った犬の1例

  田村 純, 湯本 優希, 大山 紀彦, 新坊 弦也, 滝口 満喜  北海道獣医師会雑誌  66-  (8)  289  -289  2022/08
• 日本のペット動物における有機フッ素化合物の汚染実態

  野見山 桂, 佐藤 楓夏, 水川 葉月, 池中 良徳, 田上 瑠美, 横山 望, 滝口 満喜  環境化学物質3学会合同大会要旨集  2022年-  195  -196  2022/06
• 門脈静脈連結部での選択的結紮に術中超音波ガイドが有用であった右肝区域肝内門脈体循環短絡症の犬の1例

  松本創, 細谷謙次, 細谷謙次, 大脇稜, 大山紀彦, 竹内恭介, 木之下怜平, 金尚昊, 滝口満喜, 奥村正裕  日本獣医麻酔外科学会学術集会抄録集(Web)  105th-  2022
• 「新型コロナ感染症(COVID-19)下での参加型臨床実習の実施 2020年度北海道大学動物医療センターにおける参加型臨床実習の実際

  滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSS  -2]  2021/09
• メタボロミクスおよび甲状腺ホルモン分析による有機ハロゲン汚染物質への曝露によって引き起こされたペット猫(Felis silvestris catus)の健康影響評価

  野見山 桂, 山本 恭央, 江口 哲史, 西川 博之, 水川 葉月, 横山 望, 市居 修, 滝口 満喜, 中山 翔太, 池中 良徳, 石塚 真由美, 国末 達也  環境ホルモン学会研究発表会要旨集  23回-  55  -55  2021/09
• 「新型コロナ感染症(COVID-19)下での参加型臨床実習の実施 2020年度北海道大学動物医療センターにおける参加型臨床実習の実際

  滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSS  -2]  2021/09
• 犬の僧帽弁疾患における2D Speckle Tracking心エコー図法により推定した左心房stiffnessと予後の関係

  大菅 辰幸, 黒田 杏子, 森田 智也, 中村 健介, 佐々木 東, 滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSO  -8]  2021/09
• Comprehensive analysis of steroid profile in dogs with Cushing' syndrome and hepatocellular carcinoma(和訳中)

  Oo Thandar, 佐々木 東, 池中 良徳, 永田 矩之, 一瀬 貴大, Bin Teoh Yong, 川元 誠, 小島 一輝, 菅原 芽伊, 中山 翔太, 石塚 真由美, 中村 健介, 滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSO  -18]  2021/09
• イヌの肝細胞腫瘍におけるDNAメチル化の網羅的解析

  淺利 友, 山崎 淳平, Oo Thander, 細谷 謙次, 金 尚昊, 木之下 怜平, 竹内 恭介, 賀川 由美子, 佐々木 東, 中村 健介, 滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSO  -30]  2021/09
• Identification of pathogenic Single Nucleotide Polymorphism in Inflammatory Colorectal Polyps of Miniature Dachshunds(和訳中)

  Bin Teoh Yong, 永田 矩之, 大田 寛, 佐々木 東, 中村 健介, 村上 正晃, 滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSO  -32]  2021/09
• 非再生性免疫介在性貧血の犬における脾臓摘出術前後の血清中蛋白の網羅的解析

  菅原 芽伊, 山崎 淳平, 森下 啓太郎, 金 尚昊, 細谷 謙次, 佐々木 東, 中村 健介, 滝口 満喜  日本獣医学会学術集会講演要旨集  164回-  [HSO  -60]  2021/09
• 重度肝線維化を伴う好酸球性肉芽腫を認めた好酸球増多症候群の若齢小型犬の1例

  篠原 正義, 田村 昌大, 金 尚昊, 賀川 由美子, 滝口 満喜  北海道獣医師会雑誌  65-  (8)  262  -262  2021/08
• 長期生存している特発性被嚢性腹膜硬化症の猫の1例

  前田 晴香, 横山 望, 木之下 怜平, 大田 寛, 岡田 一喜, 賀川 由美子, 滝口 満喜  北海道獣医師会雑誌  65-  (8)  263  -263  2021/08
• 左室壁肥厚と動的流出路閉塞を呈した犬の3例

  湯本 優希, 中村 健介, 志波 一恵, 寺澤 元子, 早坂 郁人, 滝口 満喜  北海道獣医師会雑誌  65-  (8)  263  -263  2021/08
• 重度肝線維化を伴う好酸球性肉芽腫を認めた好酸球増多症候群の若齢小型犬の１例

  篠原正義, 田村昌大, 金 尚昊, 賀川由美子, 滝口満喜  北海道獣医師会雑誌  65-  (8)  46  -46  2021/08
• 非造影4D-MRAにより診断した鎖骨下動脈盗血症候群の犬の1症例

  新坊弦也, 越後良介, 越後良介, 滝口満喜  北海道獣医師会雑誌  65-  (8)  268  -268  2021/08
• 超音波で生体の壁を超える 超音波と気泡を併用する膀胱内注入療法の犬における安全性検討

  佐々木 東, 青柳 貞一郎, 池中 良徳, 工藤 信樹, 中村 健介, 滝口 満喜  超音波医学  48-  (Suppl.)  S195  -S195  2021/04
• 超音波で生体の壁を超える 超音波と気泡を併用する膀胱内注入療法の犬における安全性検討

  佐々木 東, 青柳 貞一郎, 池中 良徳, 工藤 信樹, 中村 健介, 滝口 満喜  超音波医学  48-  (Suppl.)  S195  -S195  2021/04
• Analysis of variation of serum metabolites in cats by BDE209 long-term exposure test using Smart Metabolite Database

  野見山桂, 林多聞, 江口哲史, 水川葉月, 高口倖暉, 横山望, 市居修, 滝口満喜, 笹岡一慶, 中山翔太, 池中良徳, 石塚真由美, 国末達也  環境化学討論会要旨集(CD-ROM)  29th-  2021
• Elucidation of the pathophysiology of hyperadrenocortical hyperfunction (Cushing’s syndrome) by simultaneous analysis of steroid hormones using LC-MS/MS

  一瀬貴大, 安田淑一, 佐々木東, 滝口満喜, 中山翔太, 石塚真由美, 池中良徳  JSBMS Letters  46-  (Supplement)  75  -75  2021
• メラトニンの経口投与が著効した両側性副腎腫大を伴う全身性脱毛症のフェレットの1例

  BIN Teoh Yong, 亀山健吾, 永田矩之, 佐々木東, 中村健介, 滝口満喜  北海道獣医師会雑誌  65-  (8)  265  -265  2021
• Safety aspects of ultrasound-assisted intravesical chemotherapy in dogs

  佐々木東, 青柳貞一郎, 池中良徳, 工藤信樹, 中村健介, 滝口満喜  超音波医学 Supplement  48-  2021
• 診療力が試される症例 Vet Board QUIZ 肝・胆道系・膵外分泌 若齢で腹水を認めたボーダー・コリー

  田村 昌大, 滝口 満喜, 中村 健介  Veterinary Board  2-  (12)  64  -69  2020/12
• 最近の超音波による癌治療 その可能性と課題

  近藤 隆, 鈴木 亮, 佐々木 東, 滝口 満喜  Thermal Medicine  36-  (3)  75  -90  2020/09  

  超音波の作用は熱作用と非熱作用に大別され、非熱作用はキャビテーション作用と非キャビテーション作用に分けられる。以前、超音波による癌治療としては、1)超音波単独(主にハイパーサミア治療)、2)放射線治療との併用、および3)化学療法との併用があった。近年、超音波に関する生物学・基礎医学に関して、細胞死、遺伝子発現、遺伝子導入、化学物質との相互作用、薬物送達等の研究が進展してきた。とりわけ、微小気泡は診断のみならず治療のツールとして注目され、その研究は大きく発展してきた。臨床応用も集束超音波治療、低強度パルス波超音波治療、加えて血液-脳-関門の開口に関する技術の進展が著しい。本総説では、超音波による癌治療に関して、歴史的経緯を述べるとともに最近の熱作用および非熱作用を利用した癌治療についてその可能性と課題について考察する。(著者抄録)
• 非再生性貧血のミニチュア・ダックスフンド11例に対する脾臓摘出術の治療成績

  菅原 芽伊, 森下 啓太郎, 今井 健友, 山崎 淳平, 佐々木 東, 大田 寛, 細谷 謙次, 滝口 満喜  SA Medicine  22-  (3)  74  -75  2020/06
• イソフルラン麻酔下においてノルアドレナリンが全身循環に及ぼす影響

  小島 一輝, 石塚 友人, 佐藤 敬近, 滝口 満喜  日本獣医麻酔外科学雑誌  50-  (Suppl.2)  285  -285  2019/12
• Two-dimensional shear wave elastographyによる非侵襲的な犬の肝線維化診断

  田村 昌大, 大田 寛, 大菅 辰幸, 新坊 弦也, 佐々木 東, 森下 啓太郎, 賀川 由美子, 滝口 満喜  日本獣医麻酔外科学雑誌  50-  (Suppl.2)  315  -315  2019/12
• Two-dimensional shear wave elastographyによる非侵襲的な犬の肝線維化診断

  田村 昌大, 大田 寛, 大菅 辰幸, 新坊 弦也, 佐々木 東, 森下 啓太郎, 賀川 由美子, 滝口 満喜  日本獣医麻酔外科学雑誌  50-  (Suppl.2)  315  -315  2019/12  [Not refereed][Not invited]
  
• LC‐MS/MSを用いたイヌ尿中メタネフリンおよびノルメタネフリン測定

  井上裕美子, 佐々木東, 池中良徳, 一瀬貴大, 笹岡一慶, 大菅辰幸, 中山翔太, 森下啓太郎, 大田寛, 石塚真由美, 滝口満喜  日本獣医学会学術集会講演要旨集  162nd-  470  2019/08/20  [Not refereed][Not invited]
  
• LC‐MS/MS法を用いたイヌ血清中ステロイドホルモンの分析法の確立と内分泌疾患モデルを用いた実測

  白尾大樹, 池中良徳, 一瀬貴大, 中山翔太, 佐々木東, 永田矩之, 水川葉月, 滝口満喜, 石塚真由美  日本獣医学会学術集会講演要旨集  162nd-  470  2019/08/20  [Not refereed][Not invited]
  
• 【部位別の基本描出シリーズ 循環器編(3)心臓 各論 後天性疾患】超音波検査

  大菅 辰幸, 滝口 満喜  伴侶動物画像診断  4-  (4)  28  -71  2019/08
• LC-MS/MSを用いたイヌ尿中メタネフリンおよびノルメタネフリン測定

  井上 裕美子, 佐々木 東, 池中 良徳, 一瀬 貴大, 笹岡 一慶, 大菅 辰幸, 中山 翔太, 森下 啓太郎, 大田 寛, 石塚 真由美, 滝口 満喜  日本獣医学会学術集会講演要旨集  162回-  470  -470  2019/08
• LC-MS/MS法を用いたイヌ血清中ステロイドホルモンの分析法の確立と内分泌疾患モデルを用いた実測

  白尾 大樹, 池中 良徳, 一瀬 貴大, 中山 翔太, 佐々木 東, 永田 矩之, 水川 葉月, 滝口 満喜, 石塚 真由美  日本獣医学会学術集会講演要旨集  162回-  470  -470  2019/08
• 北大附属動物病院における薬剤耐性モニタリング調査結果2018

  笹岡 一慶, 森下 啓太郎, 細谷 謙次, 滝口 満喜  北海道獣医師会雑誌  63-  (8)  334  -334  2019/08
• ネコのアクロメガリー関連インスリン抵抗性糖尿病のコントロールを目的とした成長ホルモン受容体/Fc融合タンパク質の作製

  鈴木 理紗, 和泉 雄介, 前川 直也, 今内 覚, 高木 哲, 森下 啓太郎, 細谷 謙次, 滝口 満喜  日本獣医学会学術集会講演要旨集  162回-  469  -469  2019/08
• 部位別の基本描出シリーズ 循環器編 3)心臓 各論 後天性心疾患

  大菅辰幸, 滝口満喜  伴侶動物画像診断  4-  (4)  28  -71  2019/08  [Not refereed][Invited]
  
• 詳細な病理学的解析に基づいて治療方針を決定した免疫複合体性糸球体腎炎の犬の1例

  新井田 篤, 大田 寛, 笹岡 一慶, 細谷 謙次, 市居 修, 賀川 由美子, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  63-  (8)  336  -336  2019/08  [Not refereed][Not invited]
  
• 過去3年間の輸血後副反応に関する回顧的研究

  細田 千尋, 森下 啓太郎, 山崎 淳平, 高田 健介, 細谷 謙次, 金 尚昊, 佐々木 東, 大田 寛, 滝口 満喜  北海道獣医師会雑誌  63-  (8)  337  -337  2019/08  [Not refereed][Not invited]
  
• 経皮的ドレナージで良好に経過した胃排出障害を伴う膵仮性嚢胞の犬の1例

  坂本 暁彦, 大田 寛, 杉浦 岳, 康井 翔平, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  63-  (8)  341  -341  2019/08  [Not refereed][Not invited]
  
• 肝皮症候群を疑い血漿アミノ酸解析を行った肝硬変の犬の1例

  上條 太貴, 大田 寛, 賀川 由美子, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  63-  (8)  341  -341  2019/08  [Not refereed][Not invited]
  
• 犬の先天性肝外門脈体循環シャントの術後合併症率と危険因子に関する回顧的研究

  鈴木 理紗, 細谷 謙次, 金 尚昊, 森下 啓太郎, 大田 寛, 滝口 満喜, 奥村 正裕  北海道獣医師会雑誌  63-  (8)  349  -349  2019/08  [Not refereed][Not invited]
  
• 犬の僧帽弁疾患における2D Speckle Tracking心エコー図法を用いた左心房stiffnessの推定

  黒田 杏子, 大菅 辰幸, 笹岡 一慶, 森下 啓太郎, 佐々木 東, 大田 寛, 滝口 満喜  日本獣医学会学術集会講演要旨集  162回-  464  -464  2019/08  [Not refereed][Not invited]
  
• 猫の新規炎症マーカー、ロイシンリッチα2糖蛋白質(LRG)の測定系構築の試み

  坂本 暁彦, 大田 寛, 好井 健太朗, 矢吹 悠久美, 笹岡 一慶, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  日本獣医学会学術集会講演要旨集  162回-  475  -475  2019/08  [Not refereed][Not invited]
  
• PCBs曝露によるネコ肝トランスクリプトームへの影響

  水川葉月, NGYUEN Hoa Thanh, 岩田久人, 野見山桂, 池中良徳, 中山翔太, 横山望, 市居修, 滝口満喜, KHIDKHAN Kraisiri, 田辺信介, 石塚真由美  環境化学討論会要旨集(CD-ROM)  28th-  ROMBUNNO.1C‐08  2019/06/11  [Not refereed][Not invited]
  
• デカブロモジフェニルエーテル(BDE209)の長期曝露がイエネコへ及ぼす影響

  田中啓介, 野見山桂, 水川葉月, 水川葉月, 高口倖暉, 田上瑠美, 横山望, 市居修, 滝口満喜, 笹岡一慶, 中山翔太, 池中良徳, 石塚真由美, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  28th-  ROMBUNNO.1C‐09  2019/06/11  [Not refereed][Not invited]
  
• 【部位別の基本描出シリーズ 循環器編(2)心臓 各論 先天性疾患】超音波検査

  大菅 辰幸, 滝口 満喜  伴侶動物画像診断  4-  (3)  24  -66  2019/06
• 部位別の基本描出シリーズ 循環器編 2)心臓 各論 先天性心疾患

  大菅辰幸, 滝口満喜  伴侶動物画像診断  4-  (3)  24  -66  2019/06  [Not refereed][Invited]
  
• 慢性僧帽弁疾患犬において心室間流入時間差は有用な心機能評価指標である

  森田 智也, 中村 健介, 大菅 辰幸, 滝口 満喜  日本獣医麻酔外科学雑誌  50-  (Suppl.1)  322  -322  2019/06  [Not refereed][Not invited]
  
• 閉塞性黄疸はShear Wave Elastographyによる肝硬度測定に影響を与える

  田村 昌大, 大田 寛, 新坊 弦也, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  日本獣医麻酔外科学雑誌  50-  (Suppl.1)  333  -333  2019/06  [Not refereed][Not invited]
  
• 大学動物病院の活動の現状とさらなる発展を目指して 北海道大学動物医療センターの取組みと課題

  滝口 満喜  日本獣医師会雑誌  72-  (5)  240  -242  2019/05
• 蚕白喪失性腸症の犬40例からの腸リンパ管拡張症の診断と治療に関する考察

  永田 矩之, 大田 寛, 横山 望, 佐々木 東, 大菅 辰幸, 森下 啓太郎, 滝口 満喜  日本獣医師会雑誌  72-  (4)  234  -235  2019/04
• 【部位別の基本描出シリーズ 循環器編(1) 心臓 総論】超音波検査

  大菅 辰幸, 滝口 満喜  伴侶動物画像診断  4-  (2)  27  -68  2019/04
• 部位別の基本描出シリーズ 循環器編 1)心臓 総論 超音波検査

  大菅辰幸, 滝口満喜  伴侶動物画像診断  4-  (2)  27  -68  2019/04  [Not refereed][Invited]
  
• 蚕白喪失性腸症の犬40例からの腸リンパ管拡張症の診断と治療に関する考察

  永田 矩之, 大田 寛, 横山 望, 佐々木 東, 大菅 辰幸, 森下 啓太郎, 滝口 満喜  日本獣医師会雑誌  72-  (4)  234  -235  2019/04  [Not refereed][Not invited]
  
• 低強度パルス超音波照射による毛髪細胞刺激

  佐々木 東, 工藤 信樹, 大田 寛, 滝口 満喜  超音波医学  46-  (Suppl.)  S579  -S579  2019/04  [Not refereed][Not invited]
  
• 蛋白喪失性腸症の犬40例からの腸リンパ管拡張症の診断と治療に関する考察

  永田矩之, 大田寛, 横山望, 佐々木東, 大菅辰幸, 森下啓太郎, 滝口満喜  日本獣医師会獣医学術学会年次大会講演要旨集  2018-  2019
• 慢性僧帽弁疾患犬において心室間流入時間差は有用な心機能評価指標である

  森田智也, 中村健介, 大菅辰幸, 大菅辰幸, 滝口満喜  日本獣医麻酔外科学雑誌(Web)  50-  (Supplement 1)  2019
• ピモベンダンは慢性塞栓性肺高血圧症において圧を上げずに拍出量を上げる

  森田 智也, 中村 健介, 大菅 辰幸, 滝口 満喜  日本獣医麻酔外科学雑誌  49-  (Suppl.2)  242  -242  2018/12  [Not refereed][Not invited]
  
• 犬の僧帽弁疾患における心エコー図法により推定した左心室-動脈カップリングとステージの関係

  大菅 辰幸, 森田 智也, 佐々木 東, 森下 啓太郎, 大田 寛, 滝口 満喜  日本獣医麻酔外科学雑誌  49-  (Suppl.2)  249  -249  2018/12  [Not refereed][Not invited]
  
• Shear Wave Elastographyによる犬の肝硬度評価の試み

  田村 昌大, 森下 啓太郎, 新坊 弦也, 佐々木 東, 大田 寛, 滝口 満喜  日本獣医画像診断学会抄録  64回-  13  -13  2018/11
• やっぱり難しい膵炎診断 症状、膵リパーゼ、画像をどう評価するか? 膵炎の画像診断を中心に

  滝口 満喜  動物臨床医学会年次大会プロシーディング  39回-  (1)  77  -77  2018/11
• 超音波造影を含めた超音波検査の将来

  滝口 満喜  日本獣医画像診断学会抄録  64回-  9  -9  2018/11
• Shear Wave Elastographyによる犬の肝硬度評価の試み

  田村 昌大, 森下 啓太郎, 新坊 弦也, 佐々木 東, 大田 寛, 滝口 満喜  日本獣医画像診断学会抄録  64回-  13  -13  2018/11  [Not refereed][Not invited]
  
• 犬の非再生性免疫介在性貧血の臨床病理所見と治療反応に関する回顧的研究

  岩永優斗, 森下啓太郎, 細谷謙次, 大菅辰幸, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  62-  (8)  334  2018/08/24  [Not refereed][Not invited]
  
• シスプラチン膀胱内ソノポレーションの前臨床試験的検討

  佐々木東, 中山翔太, 大田寛, 大菅辰幸, 森下啓太郎, 水川葉月, 池中良徳, 石塚真由美, 滝口満喜  日本獣医学会学術集会講演要旨集  161st-  429  2018/08/21  [Not refereed][Not invited]
  
• 犬の非再生性免疫介在性貧血の臨床病理所見と治療反応に関する回顧的研究

  岩永 優斗, 森下 啓太郎, 細谷 謙次, 大菅 辰幸, 佐々木 東, 大田 寛, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  334  -334  2018/08
• シスプラチン膀胱内ソノポレーションの前臨床試験的検討

  佐々木 東, 中山 翔太, 大田 寛, 大菅 辰幸, 森下 啓太郎, 水川 葉月, 池中 良徳, 石塚 真由美, 滝口 満喜  日本獣医学会学術集会講演要旨集  161回-  429  -429  2018/08
• 術前からダイナミックな血圧変動を呈した褐色細胞腫を良好に麻酔管理した犬の1例

  小島 一輝, 石塚 友人, 金 尚昊, 竹内 恭介, 細谷 謙次, 滝口 満喜, 奥村 正裕  北海道獣医師会雑誌  62-  (8)  336  -336  2018/08
• 蛋白喪失性腸症の犬33例からの腸リンパ管拡張症の診断と治療に関する考察

  永田 矩之, 大田 寛, 横山 望, ニサ・ホイルン, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  334  -334  2018/08  [Not refereed][Not invited]
  
• 神経筋接合部検査としてアイスパックテストを試みた重症筋無力症の犬の1例

  山田 有紗, 笹岡 一慶, 大田 寛, 立花 由莉加, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  335  -335  2018/08  [Not refereed][Not invited]
  
• 犬の粘液腫様変性性僧帽弁疾患における心エコー図法による左心室-大動脈カップリングの推定

  大菅 辰幸, 佐々木 東, 森下 啓太郎, 大田 寛, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  335  -335  2018/08  [Not refereed][Not invited]
  
• 神経症状を主訴とした悪性中皮腫の猫の1例

  大村 侑己, 大田 寛, 小林 篤史, 笹岡 一慶, 道下 明日香, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  340  -340  2018/08  [Not refereed][Not invited]
  
• 経頭蓋超音波ドプラ法により局所的な脳循環動態を評価し得た右中大脳動脈梗塞の犬の1例

  笹岡 一慶, 大田 寛, 石塚 友人, 大菅 辰幸, 立花 由莉加, 森下 啓太郎, 佐々木 東, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  341  -341  2018/08  [Not refereed][Not invited]
  
• 鼻腔内真菌感染症に対しクロトリマゾール灌流および注入療法を実施した犬の1例

  大谷 豪, 大田 寛, 岩木 有香, 細谷 謙次, 新坊 弦也, 佐々木 東, 森下 啓太郎, 滝口 満喜  北海道獣医師会雑誌  62-  (8)  344  -344  2018/08  [Not refereed][Not invited]
  
• 健康な犬における二次元スペックルトラッキング心エコーによる左心室長軸方向ストレイン解析に与える心臓容積負荷の影響(Effect of cardiac volume loading on left atrial longitudinal strain analysis derived from two-dimensional Speckle Tracking Echocardiography in healthy dogs)

  Dermlim Angkhana, 中村 健介, 森田 智也, 大菅 辰幸, Nisa Khoirun, Leela-arporn Rommaneeya, 笹岡 一慶, 永田 矩之, 田村 昌大, 佐々木 東, 大田 寛, 森下 啓太郎, 滝口 満喜  日本獣医学会学術集会講演要旨集  161回-  435  -435  2018/08  [Not refereed][Not invited]
  
• 蛋白喪失性腸症の犬33例からの腸リンパ管拡張症の診断と治療に関する考察

  永田矩之, 大田寛, 横山望, ニサ ホイルン, 佐々木東, 森下啓太郎, 滝口満喜  北海道獣医師会雑誌  62-  (8)  334  -334  2018/08  [Not refereed][Not invited]
  
• 神経筋接合部検査としてアイスパックテストを試みた重症筋無力症の犬の1例

  山田有紗, 笹岡一慶, 大田寛, 立花由莉加, 大菅辰幸, 佐々木東, 森下啓太郎, 滝口満喜  北海道獣医師会雑誌  62-  (8)  335  -335  2018/08  [Not refereed][Not invited]
  
• 経頭蓋超音波ドプラ法により局所的な脳循環動態を評価し得た右中大脳動脈梗塞の犬の1例

  笹岡一慶, 大田寛, 石塚友人, 大菅辰幸, 立花由莉加, 森下啓太郎, 佐々木東, 滝口満喜  北海道獣医師会雑誌  62-  (8)  341  -341  2018/08  [Not refereed][Not invited]
  
• 神経症状を主訴とした悪性中皮腫の猫の1例

  大村侑己, 大田寛, 小林篤史, 笹岡一慶, 道下明日香, 大菅辰幸, 佐々木東, 森下啓太郎, 滝口満喜  北海道獣医師会雑誌  62-  (8)  340  -340  2018/08  [Not refereed][Not invited]
  
• 犬の粘液腫様変性性僧帽弁疾患における心エコー図法による左心室‐大動脈カップリングの推定

  大菅辰幸, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  北海道獣医師会雑誌  62-  (8)  335  -335  2018/08  [Not refereed][Not invited]
  
• 鼻腔内真菌感染症に対しクロトリマゾール潅流および注入療法を実施した犬の1例

  大谷豪, 大田寛, 岩木有香, 細谷謙次, 新坊弦也, 佐々木東, 森下啓太郎, 滝口満喜  北海道獣医師会雑誌  62-  (8)  344  -344  2018/08  [Not refereed][Not invited]
  
• 術前からダイナミックな血圧変動を呈した褐色細胞腫を良好に麻酔管理した犬の1例

  小島 一輝, 石塚 友人, 金 尚昊, 竹内 恭介, 細谷 謙次, 滝口 満喜, 奥村 正裕  北海道獣医師会雑誌  62-  (8)  336  -336  2018/08  [Not refereed][Not invited]
  
• Shear Wave Elastography(SWE)を用いた犬の肝硬度測定

  田村 昌大, 大田 寛, 佐々木 東, 賀川 由美子, 滝口 満喜  獣医畜産新報  71-  (7)  496  -497  2018/07  [Not refereed][Not invited]
  
• Shear Wave Elastography(SWE)を用いた犬の肝硬度測定

  田村昌大, 大田寛, 佐々木東, 賀川由美子, 滝口満喜  獣医畜産新報  71-  (7)  496  -497  2018/07  [Not refereed][Not invited]
  
• 慢性僧帽弁疾患犬において右室strainによる右心室機能評価は優れた予後指標となり得るか

  森田 智也, 中村 健介, 大菅 辰幸, 森下 啓太郎, 佐々木 東, 大田 寛, 滝口 満喜  SA Medicine  20-  (3)  68  -70  2018/06
• ポリ塩化ビフェニル曝露によるイヌ・ネコの甲状腺ホルモン恒常性への影響

  高口 倖暉, 野見山 桂, 西川 博之, 水川 葉月, 田上 瑠美, 草木 桃子, 横山 望, 市居 修, 滝口 満喜, 中山 翔太, 池中 良徳, 石塚 真由美, 岩田 久人, 国末 達也, 田辺 信介  The Journal of Toxicological Sciences  43-  (Suppl.)  S245  -S245  2018/06
• 肺塞栓症において認められる心室中隔の奇異性運動は左右心室間同期障害によって生じる

  中村健介, 中村健介, 森田智也, 大菅辰幸, 池田正悟, DERMLIM Angkhana, 滝口満喜  日本獣医麻酔外科学雑誌  49-  (Suppl.1)  301  -301  2018/06  [Not refereed][Not invited]
  
• 慢性僧帽弁疾患犬において右室strainによる右心室機能評価は優れた予後指標となり得るか

  森田智也, 中村健介, 大菅辰幸, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  SA Medicine  20-  (3)  68‐70  2018/06/01  [Not refereed][Not invited]
  
• 有機ハロゲン化合物曝露によるネコの生化学マーカーの変化および甲状腺機能に及ぼす影響評価

  水川葉月, KHIDKHAN Kraisiri, 高口倖暉, 横山望, 市居修, 滝口満喜, 笹岡一慶, 池中良徳, 池中良徳, 中山翔太, TSEND‐AYUSH Sainnoxoi, 野見山桂, 田辺信介, 石塚真由美  環境化学討論会要旨集(CD-ROM)  27th-  ROMBUNNO.2A‐10  2018/05/11  [Not refereed][Not invited]
  
• ネコのin vivo BDE209長期曝露試験による体内動態の解明および甲状腺ホルモンへの影響評価

  田中啓介, 野見山桂, 水川葉月, 高口倖暉, 田上瑠美, 横山望, 市居修, 滝口満喜, 笹岡一慶, 中山翔太, 池中良徳, 石塚真由美, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  27th-  ROMBUNNO.1A‐03(PA‐13)  2018/05/11  [Not refereed][Not invited]
  
• 犬の蛋白漏出性腸症の予後因子解析 超音波検査所見の有用性の検討

  大田 寛, 賀川 由美子, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  日本獣医師会雑誌  71-  (4)  206  -207  2018/04
• 【部位別の基本描出シリーズ 内分泌器官編 甲状腺・上皮小体】超音波検査

  滝口 満喜  伴侶動物画像診断  3-  (2)  54  -62  2018/04
• ロジカルに修得する超音波検査(第1回) 総論 超音波診断装置とその調整

  滝口 満喜  伴侶動物画像診断  3-  (2)  98  -101  2018/04
• 犬の蛋白漏出性腸症の予後因子解析 超音波検査所見の有用性の検討

  大田 寛, 賀川 由美子, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  日本獣医師会雑誌  71-  (4)  206  -207  2018/04  [Not refereed][Not invited]
  
• 低強度パルス超音波による神経軸索の伸展制御の試み

  佐々木東, 工藤信樹, 大田寛, 滝口満喜  超音波医学  45-  (Suppl.)  S575  -S575  2018/04  [Not refereed][Not invited]
  
• 血清中メタボローム解析によるペットネコ(Felis catus)の有機ハロゲン化合物毒性影響評価

  野見山桂, 江口哲史, 西川博之, 水川葉月, 山本恭央, 滝口満喜, 中山翔太, 池中良徳, 石塚真由美, 田辺信介  環境化学討論会要旨集(CD-ROM)  27th-  2018
• JCVIM 2018 臨床研究アワード 慢性僧帽弁疾患犬において右室strainによる右心室機能評価は優れた予後指標となり得るか (第14回日本獣医内科学アカデミー学術大会(JCVIM 2018) 〈臨床研究アワード〉日本獣医内科学アカデミー/インターズー学術奨励賞受賞作品ダイジェスト版)

  森田 智也, 中村 健介, 大菅 辰幸, 森下 啓太郎, 佐々木 東, 大田 寛, 滝口 満喜  SA medicine : Journal of small animal medicine  24-  (3)  68  -70  2018  [Not refereed][Not invited]
  
• Effects of PCB-exposure on thyroid hormone homeostasis in dog (canis lupus familiaris) and cat (felis catus)

  TAKAGUCHI Kohki, NAKAYAMA Shouta, IKENAKA Yoshinori, ISHIZUKA Mayumi, IWATA Hisato, KUNISUE Tatsuya, TANABE Shinsuke, NOMIYAMA Kei, NISHIKAWA Hiroyuki, MIZUKAWA Hazuki, TANOUE Rumi, KUSAKI Momoko, YOKOYAMA Nozomu, ICHII Osamu, TAKIGUCHI Mitsuyoshi  Annual Meeting of the Japanese Society of Toxicology  45-  (0)  P  -72  2018  [Not refereed][Not invited]
   

  <p>[in Japanese] </p>
• 内分泌性脱毛症の2例

  島崎 博美, 飯田 大起, 佐々木 東, 滝口 満喜, 藤井 義晴  (一社)比較統合医療学会学術大会・日本補完代替医療学会学術集会プログラム・抄録集  60回・20回-  57  -58  2017/12
• ポリ塩化ビフェニル(PCBs)がネコの甲状腺機能に及ぼす影響

  水川 葉月, 高居 名菜, 野見山 桂, 高口 倖暉, 横山 望, 市居 修, 滝口 満喜, 池中 良徳, 中山 翔太, 石塚 真由美  環境ホルモン学会研究発表会要旨集  20回-  78  -78  2017/12
• ポリ塩化ビフェニル(PCBs)がネコの甲状腺機能に及ぼす影響

  水川葉月, 高居名菜, 野見山桂, 高口倖暉, 横山望, 市居修, 滝口満喜, 池中良徳, 池中良徳, 中山翔太, 石塚真由美  日本内分泌かく乱化学物質学会研究発表会要旨集  20th-  78  -78  2017/12  [Not refereed][Not invited]
  
• 消化管エコーのノウハウ

  滝口 満喜  動物臨床医学会年次大会プロシーディング  38回-  (1)  91  -91  2017/11
• すぐに診療で活かせる超音波画像診断の臨床

  滝口 満喜  動物臨床医学会年次大会プロシーディング  38回-  (3)  205  -205  2017/11
• 生前診断に苦慮した化膿性腸炎の猫の1例

  森下 啓太郎, 中島 彩乃, 賀川 由美子, 華園 究, 佐々木 東, 大田 寛, 滝口 満喜  日本獣医画像診断学会抄録  62回-  6  -6  2017/09
• 生前診断に苦慮した化膿性腸炎の猫の1例

  森下啓太郎, 中島彩乃, 賀川由美子, 華園究, 佐々木東, 大田寛, 滝口満喜  日本獣医画像診断学会抄録(Web)  62nd-  6 (WEB ONLY)  2017/09  [Not refereed][Not invited]
  
• 超音波と膀胱内注入療法を組み合わせた膀胱癌治療法の開発

  佐々木東, 中山翔太, 森下啓太郎, 中村健介, 大田寛, 水川葉月, 池中良徳, 石塚真由美, 滝口満喜  日本獣医学会学術集会講演要旨集  160th-  461  2017/08/30  [Not refereed][Not invited]
  
• 犬の右肝区域におけるCT画像解析を用いた静脈血管走行の多様性調査

  西野綾乃, 華園究, 石塚友人, 星野有希, 森下啓太郎, 滝口満喜, 高木哲  日本獣医学会学術集会講演要旨集  160th-  455  2017/08/30  [Not refereed][Not invited]
  
• 犬の右肝区域におけるCT画像解析を用いた静脈血管走行の多様性調査

  西野 綾乃, 華園 究, 石塚 友人, 星野 有希, 森下 啓太郎, 滝口 満喜, 高木 哲  日本獣医学会学術集会講演要旨集  160回-  455  -455  2017/08
• 超音波と膀胱内注入療法を組み合わせた膀胱癌治療法の開発

  佐々木 東, 中山 翔太, 森下 啓太郎, 中村 健介, 大田 寛, 水川 葉月, 池中 良徳, 石塚 真由美, 滝口 満喜  日本獣医学会学術集会講演要旨集  160回-  461  -461  2017/08
• 造影超音波検査により計測する肺通過時間は肺動脈塞栓モデル犬において短縮する

  中村 健介, 森田 智也, 大菅 辰幸, 滝口 満喜  日本獣医学会学術集会講演要旨集  160回-  465  -465  2017/08  [Not refereed][Not invited]
  
• 副腎偶発腫から診断した犬のサブクリニカルクッシング症候群の臨床的特徴

  永田矩之, 大田寛, 佐々木東, 森下啓太郎, 中村康介, 中村康介, 細谷謙次, 高木哲, 滝口満喜  北海道獣医師会雑誌  61-  (8)  326  -326  2017/08  [Not refereed][Not invited]
  
• 組織球性大腸炎のフレンチ・ブルドッグの1例

  立花由莉加, 大田寛, 賀川由美子, 佐々木東, 森下啓太郎, 滝口満喜  北海道獣医師会雑誌  61-  (8)  348  -348  2017/08  [Not refereed][Not invited]
  
• 後天性門脈体循環シャントを合併した先天性肝線維症の猫の1例

  田村昌大, 森下啓太郎, 高木哲, 早川小百合, 賀川由美子, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  61-  (8)  345  -345  2017/08  [Not refereed][Not invited]
  
• 市販フードの給餌により栄養性二次性上皮小体機能亢進症を生じた犬の2例

  道下明日香, 森下啓太郎, 橋場香野, 田村昌大, 金尚昊, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  61-  (8)  327  -327  2017/08  [Not refereed][Not invited]
  
• 犬の蛋白漏出性腸症の予後因子解析:超音波検査所見の有用性の検討

  大田寛, 賀川由美子, 佐々木東, 森下啓太郎, 滝口満喜  北海道獣医師会雑誌  61-  (8)  347  -347  2017/08  [Not refereed][Not invited]
  
• 犬の原発性免疫介在性血小板減少症(pIMT)再発症例の回顧的研究

  高村大地, 森下啓太郎, 中島彩乃, 竹内恭介, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  61-  (8)  332  -332  2017/08  [Not refereed][Not invited]
  
• 犬慢性腸症の十二指腸粘膜におけるロイシンリッチα2糖蛋白の発現とその臨床的意義

  奥朋哉, 大田寛, 田村昌大, 永田矩之, 笹岡一慶, LEELA‐ARPORN Rommaneeya, DERMLIM Angkhana, NISA Khoirun, 森田智也, 大菅辰幸, 佐々木東, 森下啓太郎, 滝口満喜  日本獣医学会学術集会講演要旨集  160回-  477  -477  2017/08  [Not refereed][Not invited]
  
• 全エクソームシーケンスを用いたミニチュア・ダックスフントの炎症性結直腸ポリープ感受性遺伝子の探索

  永田矩之, 大田寛, 横山望, 田村昌大, 笹岡一慶, LEELA‐ARPORN Rommaneeya, DERMLIM Angkhana, NISA Khoirun, 森田智也, 大菅辰幸, 佐々木東, 森下啓太郎, 滝口満喜  日本獣医学会学術集会講演要旨集  160回-  477  -477  2017/08  [Not refereed][Not invited]
  
• 犬虚血性脳血管障害における脳底動脈血流の経頭蓋超音波ドプラ法による評価

  笹岡一慶, 大田寛, 田村昌大, 永田矩之, LEELA‐ARPORN Rommaneeya, DERMLIM Angkhana, KHOIRUN Nisa, 森田智也, 佐々木東, 森下啓太郎, 滝口満喜  日本獣医学会学術集会講演要旨集  160回-  465  -465  2017/08  [Not refereed][Not invited]
  
• 慢性肺動脈塞栓性肺高血圧症モデル犬を用いたピモベンダンの作用に関する検討

  森田智也, 中村健介, 大菅辰幸, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  日本獣医学会学術集会講演要旨集  160回-  465  -465  2017/08  [Not refereed][Not invited]
  
• 慢性腸疾患の犬における十二指腸の定量的造影超音波法(Quantitative contrast-enhanced ultrasonography of duodenum in dogs with chronic enteropathy)

  Khoirun Nisa, 大田 寛, 横山 望, 田村 昌大, 永田 矩之, 笹岡 一慶, Dermlim Angkhana, Leela-Arporn Rommaneeya, 森田 智也, 大菅 辰幸, 佐々木 東, 森下 啓太郎, 滝口 満喜  日本獣医学会学術集会講演要旨集  160回-  477  -477  2017/08  [Not refereed][Not invited]
  
• ソノポレーションを用いた膀胱内注入療法の開発 (特集 進化する超音波治療技術の最前線(1))

  佐々木 東, 工藤 信樹, 中山 翔太, 滝口 満喜  超音波techno  29-  (4)  27  -30  2017/07  [Not refereed][Not invited]
  
• ポリ塩化ビフェニル(PCBs)暴露によるネコ甲状腺機能への影響評価~甲状腺ホルモンの脱ヨード化と代謝能の解析~

  水川葉月, 高居名菜, 横山望, 市居修, 滝口満喜, 野見山桂, 高口倖暉, 西川博之, 池中良徳, 池中良徳, 中山翔太, SAINNOXOI Tsend‐ayush, 田辺信介, 石塚真由美  環境化学討論会要旨集(CD-ROM)  26th-  ROMBUNNO.2A‐01  2017/06/05  [Not refereed][Not invited]
  
• PCBs曝露がイヌ・ネコの甲状腺ホルモン恒常性へ及ぼす影響

  高口倖暉, 野見山桂, 西川博之, 水川葉月, 田上瑠美, 芳之内結加, 横山望, 市居修, 滝口満喜, 中山翔太, 池中良徳, 石塚真由美, 岩田久人, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  26th-  ROMBUNNO.2A‐02  2017/06/05  [Not refereed][Not invited]
  
• イエネコのin vivo長期曝露試験:デカブロモジフェニルエーテル(BDE209)は甲状腺機能亢進症を引き起こすのか?

  水川葉月, 野見山桂, 池中良徳, 池中良徳, 中山翔太, SAENGTIENCHAI Aksorn, SAINNOXOI Tsendayush, 横山望, 笹岡一慶, 市居修, KHIDKHAN Kraisiri, 高居名菜, 滝口満喜, 田辺信介, 石塚真由美  環境化学討論会要旨集(CD-ROM)  26th-  ROMBUNNO.2A‐03  2017/06/05  [Not refereed][Not invited]
  
• イエネコのin vivo長期曝露試験:デカブロモジフェニルエーテル(BDE209)の生体内変化

  田中啓介, 野見山桂, 水川葉月, 高口倖暉, 田上瑠美, SAENGTIENCHAI Aksorn, SAINNOXOI Tsend‐ayush, 横山望, 市居修, 滝口満喜, 中山翔太, 池中良徳, 石塚真由美, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  26th-  ROMBUNNO.2A‐04  2017/06/05  [Not refereed][Not invited]
  
• 血清中メタボローム解析によるイエネコのPCBs毒性影響評価

  野見山桂, 西川博之, 水川葉月, 高口倖暉, SAINNOXOI Tsend‐ayush, 横山望, 市居修, 滝口満喜, 石塚真由美, 池中良徳, 中山翔太, 江口哲史, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  26th-  ROMBUNNO.2A‐05  2017/06/05  [Not refereed][Not invited]
  
• ネコの甲状腺機能に対するポリ塩化ビフェニル(PCBs)の影響評価 甲状腺ホルモンの脱ヨード化と代謝能の解析

  水川 葉月, 高居 名菜, 横山 望, 市居 修, 滝口 満喜, 野見山 桂, 高口 倖暉, 西川 博之, 池中 良徳, 中山 翔太, Sainnoxoi Tsend-ayush, 田辺 信介, 石塚 真由美  The Journal of Toxicological Sciences  42-  (Suppl.)  S236  -S236  2017/06
• 正常犬ならびに慢性僧帽弁疾患犬におけるピモベンダン投与による左心房機能の変化

  三木伸悟, 中村健介, 大菅辰幸, 森田智也, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  48-  (Suppl.1)  348  -348  2017/06  [Not refereed][Not invited]
  
• 急性圧負荷が右室にもたらす影響:心エコー図検査は何を反映しているか

  森田智也, 中村健介, 大菅辰幸, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  48-  (Suppl.1)  336  -336  2017/06  [Not refereed][Not invited]
  
• 先天性門脈体循環シャントに伴う肝肺症候群および門脈肺高血圧症を疑った1例

  道下明日香, 中村健介, 森下啓太郎, 佐々木東, 大田寛, 細谷謙次, 賀川由美子, 滝口満喜  日本獣医麻酔外科学雑誌  48-  (Suppl.1)  220  -220  2017/06  [Not refereed][Not invited]
  
• ネコの甲状腺機能に対するポリ塩化ビフェニル(PCBs)の影響評価~甲状腺ホルモンの脱ヨード化と代謝能の解析~

  水川葉月, 高居名菜, 横山望, 市居修, 滝口満喜, 野見山桂, 高口倖暉, 西川博之, 池中良徳, 池中良徳, 中山翔太, SAINNOXOI Tsend‐Ayush, 田辺信介, 石塚真由美  Journal of Toxicological Sciences  42-  (Supplement)  S236  2017/06  [Not refereed][Not invited]
   

  有機ハロゲン化合物の一種であるポリ塩化ビフェニル（PCBs）は甲状腺ホルモン（THs）の恒常性かく乱が懸念されており、実際、ヒトやげっ歯類においてはPCBs暴露により血中THs濃度の低下が認められている。ヒトの身近な動物であるネコは餌などを介してPCBsに暴露していると懸念され、THs恒常性に何らかの悪影響をもたらすことが予想される。本研究では、ネコの甲状腺機能に対するPCBsの影響を解析するため、in vivo投与試験したネコを用いて甲状腺機能への影響を解析した。血中のTHs濃度（総T4、総T3、遊離T4、遊離T3、リバースT3）を解析した結果、対照群と暴露群の間に有意な差は見られなかった。また、THsの恒常性に関与する遺伝子（甲状腺ホルモン受容体αおよびβ、脱ヨード酵素type1）の肝臓中発現量やTHs代謝に関わる酵素であるグルクロン酸抱合酵素（UGT1A1）の活性を調べたところ、対照群と暴露群の間に有意な差は見られなかった。
  本実験結果では、ネコにおいてPCBs短期暴露によるTHsへの影響が認められなかったが、日本のペットネコにおけるモニタリング調査では、血中のTHs濃度とPCBs濃度との間に負の相関が見られており、有機ハロゲン化合物によるTHsかく乱が懸念されている。また、本研究試料のメタボローム解析結果では酸化ストレスの発現、抗酸化物質、電子伝達物質の生産阻害などが示唆されており、PCBsや水酸化代謝物による内分泌撹乱作用が心配される。THsの恒常性は非常に複雑な機構のもと維持されていることが知られており、負のフィードバック機構に加え、THsの活性化や代謝、排泄の調整にも多くの遺伝子やトランスポーター、酵素など様々な要因が関わっていることから、ネコは長期的かつ慢性的に化学物質に暴露されることで、甲状腺機能などへの悪影響が現れると考えられ、内分泌撹乱作用の作用機序について解析を進めることが重要である。
• Thyroid hormone effects of polychlorinated biphenyls (PCBs) in cats-Analysis of deiodination and metabolism-

  MIZUKAWA Hazuki, NAKAYAMA Shouta M.M, SAINNOXOI Tsend-ayush, TANABE Shinsuke, ISHIZUKA Mayumi, TAKAI Nana, YOKOYAMA Nozomu, ICHII Osamu, TAKIGUCHI Mitsuyoshi, NOMIYAMA Kei, TAKAGUCHI Kohki, NISHIKAWA Hiroyuki, IKENAKA Yoshinori  Annual Meeting of the Japanese Society of Toxicology  44-  (0)  P  -55  2017/06  [Not refereed][Not invited]
   

  有機ハロゲン化合物の一種であるポリ塩化ビフェニル（PCBs）は甲状腺ホルモン（THs）の恒常性かく乱が懸念されており、実際、ヒトやげっ歯類においてはPCBs暴露により血中THs濃度の低下が認められている。ヒトの身近な動物であるネコは餌などを介してPCBsに暴露していると懸念され、THs恒常性に何らかの悪影響をもたらすことが予想される。本研究では、ネコの甲状腺機能に対するPCBsの影響を解析するため、<i>in vivo</i>投与試験したネコを用いて甲状腺機能への影響を解析した。血中のTHs濃度（総T4、総T3、遊離T4、遊離T3、リバースT3）を解析した結果、対照群と暴露群の間に有意な差は見られなかった。また、THsの恒常性に関与する遺伝子（甲状腺ホルモン受容体αおよびβ、脱ヨード酵素type1）の肝臓中発現量やTHs代謝に関わる酵素であるグルクロン酸抱合酵素（UGT1A1）の活性を調べたところ、対照群と暴露群の間に有意な差は見られなかった。<br>本実験結果では、ネコにおいてPCBs短期暴露によるTHsへの影響が認められなかったが、日本のペットネコにおけるモニタリング調査では、血中のTHs濃度とPCBs濃度との間に負の相関が見られており、有機ハロゲン化合物によるTHsかく乱が懸念されている。また、本研究試料のメタボローム解析結果では酸化ストレスの発現、抗酸化物質、電子伝達物質の生産阻害などが示唆されており、PCBsや水酸化代謝物による内分泌撹乱作用が心配される。THsの恒常性は非常に複雑な機構のもと維持されていることが知られており、負のフィードバック機構に加え、THsの活性化や代謝、排泄の調整にも多くの遺伝子やトランスポーター、酵素など様々な要因が関わっていることから、ネコは長期的かつ慢性的に化学物質に暴露されることで、甲状腺機能などへの悪影響が現れると考えられ、内分泌撹乱作用の作用機序について解析を進めることが重要である。
• 低強度パルス超音波による毛包構成細胞への刺激

  佐々木東, 工藤信樹, 中村健介, 森下啓太郎, 大田寛, 滝口満喜  再生医療  16-  348  2017/02/01  [Not refereed][Not invited]
  
• 毛包を構成する細胞に対する低強度超音波刺激

  佐々木東, 工藤信樹, 森下啓太郎, 大田寛, 滝口満喜  日本超音波医学会光超音波画像研究会プログラム・抄録集  2017-  (2nd)  66‐67  2017  [Not refereed][Not invited]
  
• 犬の甲状腺機能低下症を誤診しないように 画像診断

  滝口 満喜  動物臨床医学会年次大会プロシーディング  37回-  (1)  194  -194  2016/11
• もう一度見直す超音波検査法 基本テクニックと各種疾患の診断・評価法 腹部エコー診断がどこまで治療に役立つか

  滝口 満喜  動物臨床医学会年次大会プロシーディング  37回-  (1)  341  -341  2016/11
• Post‐capillary肺高血圧症ではPre‐capillary肺高血圧症ほど右室機能指標は減弱しない

  森田智也, 中村健介, 大菅辰幸, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  47-  (Suppl.2)  228  -228  2016/11  [Not refereed][Not invited]
  
• 3次元培養でのシスプラチンと超音波 マイクロバブルの併用

  佐々木 東, Moonen Chrit, 滝口 満喜  超音波医学  43-  (5)  673  -673  2016/09
• 超音波による細胞周期変動を利用した増感放射線療法の可能性

  藤沢 眞代, 佐々木 東, 安井 博宣, 工藤 信樹, 滝口 満喜  超音波医学  43-  (5)  679  -679  2016/09
• 膀胱癌を模した3次元培養系でのシスプラチンデリバリー

  佐々木 東, 工藤 信樹, 滝口 満喜  超音波医学  43-  (5)  679  -679  2016/09
• 3次元培養でのシスプラチンと超音波 マイクロバブルの併用

  佐々木 東, Moonen Chrit, 滝口 満喜  超音波医学  43-  (5)  673  -673  2016/09  [Not refereed][Not invited]
  
• 超音波による細胞周期変動を利用した増感放射線療法の可能性

  藤沢 眞代, 佐々木 東, 安井 博宣, 工藤 信樹, 滝口 満喜  超音波医学  43-  (5)  679  -679  2016/09  [Not refereed][Not invited]
  
• 膀胱癌を模した3次元培養系でのシスプラチンデリバリー

  佐々木 東, 工藤 信樹, 滝口 満喜  超音波医学  43-  (5)  679  -679  2016/09  [Not refereed][Not invited]
  
• 心エコー図検査にて急性期および回復後の変化を捉えた急性肺血栓塞栓症の犬の1症例

  森田智也, 中村健介, 森下啓太郎, 竹内恭介, 大菅辰幸, 那須香菜子, 華園究, 滝口満喜  北海道獣医師会雑誌  60-  (8)  417  -417  2016/08/10  [Not refereed][Not invited]
  
• 好塩基球性白血病の犬の1症例

  渡邉清正, 森下啓太郎, 中村健介, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  60-  (8)  406  -406  2016/08  [Not refereed][Not invited]
  
• 大腸血管拡張症の犬の1症例

  竹内恭介, 大田寛, 高木哲, 中島彩乃, 佐々木東, 森下啓太郎, 中村健介, 滝口満喜  北海道獣医師会雑誌  60-  (8)  418  -418  2016/08  [Not refereed][Not invited]
  
• ミニチュア・ダックスフンドの炎症性結直腸ポリープにおけるCOX‐2発現とToll‐like receptorとの関連性の解析

  長谷川純也, 大田寛, 横山望, 笹岡一慶, NISA Khoirun, 森田智也, 大菅辰幸, 佐々木東, 森下啓太郎, 中村健介, 滝口満喜  日本獣医学会学術集会講演要旨集  159回-  456  -456  2016/08  [Not refereed][Not invited]
  
• 犬の免疫介在性関節炎の疫学的および臨床的特徴

  永田 矩之, 中村 健介, 森下 啓太郎, 越後 良介, 奥村 正裕, 佐々木 東, 大田 寛, 滝口 満喜  北海道獣医師会雑誌  60-  (8)  416  -416  2016/08  [Not refereed][Not invited]
  
• ネコにおけるポリ塩化ビフェニル(PCBs)の異物代謝機構解明と甲状腺ホルモンへの影響評価

  水川葉月, 前原美咲, 横山望, 市居修, 滝口満喜, 野見山桂, 西川博之, 池中良徳, 中山翔太, 高口倖暉, 田辺信介, 石塚真由美  環境化学討論会要旨集(CD-ROM)  25th-  ROMBUNNO.1A‐13  2016/06/06  [Not refereed][Not invited]
  
• メタボローム解析を用いたイエネコにおけるPCBsの毒性影響評価

  西川博之, 野見山桂, 水川葉月, 横山望, 市居修, 滝口満喜, 石塚真由美, 池中良徳, 中山翔太, 江口哲史, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  25th-  ROMBUNNO.1A‐12  2016/06/06  [Not refereed][Not invited]
  
• 犬猫の脳疾患における造影FLAIRの使用経験

  華園究, 大田寛, 山根由久, 那須香菜子, 細谷兼次, 石塚友人, 伊丹貴晴, 滝口満喜  日本獣医麻酔外科学雑誌  47-  (Supplement 1)  335  -335  2016/06/05  [Not refereed][Not invited]
  
• ポリ塩化ビフェニル(PCBs)のin vivo暴露によるネコの異物代謝解明と甲状腺ホルモンへの影響評価

  水川 葉月, 前原 美咲, 横山 望, 市居 修, 滝口 満喜, 野見山 桂, 西川 博之, 池中 良徳, 中山 翔太, 高口 倖暉, 田辺 信介, 石塚 真由美  The Journal of Toxicological Sciences  41-  (Suppl.)  S215  -S215  2016/06
• Parametric Micro flow imaging法を用いて肝実質の動脈化を検出した門脈血栓症の犬の1例

  田村昌大, 中村健介, 森俊輔, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  47-  (Suppl.1)  335  -335  2016/06  [Not refereed][Not invited]
  
• 間質性肺疾患による重度肺高血圧症を呈したウェルシュ・コーギーの2例

  森田智也, 中村健介, 大菅辰幸, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  47-  (Suppl.1)  235  -235  2016/06  [Not refereed][Not invited]
  
• 造影超音波検査により計測した肺循環時間と慢性房室弁疾患の重症度との関係

  池田正悟, 中村健介, 大菅辰幸, 森田智也, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  47-  (Suppl.1)  323  -323  2016/06  [Not refereed][Not invited]
  
• In vivo analysis of metabolic capacities and effects on the thyroid hormone and in cats treated with polychlorinated biphenyls

  MIZUKAWA Hazuki, MAEHARA Misaki, YOKOYAMA Nozomu, ICHII Osamu, TAKIGUCHI Mitsuyoshi, NOMIYAMA Kei, NISHIKAWA Hiroyuki, IKENAKA Yoshinori, NAKAYAMA Shouta M.M, TAKAGUCHI Kohki, TANABE Shinsuke, ISHIZUKA Mayumi  Annual Meeting of the Japanese Society of Toxicology  43-  (0)  P  -6  2016/06  [Not refereed][Not invited]
   

  ポリ塩化ビフェニル（PCBs）の水酸化代謝物であるOH-PCBsは、肝臓内で薬物代謝酵素より生成され、その後体外へ排泄される。しかしながら、一部の水酸化代謝物は甲状腺ホルモン（TH）と類似の構造をもつため、THの恒常性を撹乱することが危惧されている。これまでに、多様な陸棲哺乳類の血中OH-PCBsを分析したところ、種間でOH-PCBsの組成に差異が認められ、中でも、ネコのOH-PCBs残留パターンは他種と大きく異なることから、本種は特異な代謝機能を有することが示唆された。しかし、ネコの異物代謝能の研究は僅かであり、化学物質暴露による毒性影響も不明な点が多い。本研究では、ネコにおけるPCBs <i>in vivo</i>暴露試験を実施し、体内動態および代謝に関与する酵素活性や遺伝子を解析するとともに、化学物質の暴露評価に繋がる基盤的情報の収集を目的とした。<br>コーン油に溶解した12異性体のPCBsを腹腔内投与し、経時採血した血清中PCBsおよびOH-PCBs濃度について同条件で実施したイヌの<i>in vivo</i>試験と比較した結果、異性体の残留パターンや体内動態にイヌとネコで種差が観察された。とくにネコでは低塩素化体の残留が顕著であった。また、代謝酵素活性および遺伝子解析の結果、PCBs暴露によりEROD、MROD、PROD活性は上昇するものの、第2相抱合酵素（UGTやSULT）活性は変化せず、PCBs暴露による抱合酵素活性への影響もみられなかった。また、<i>CYP1A1</i>および<i>CYP1A2</i>遺伝子の発現量の上昇も認められた。<br> 本研究により、ネコのPCBs吸収・代謝・排泄能はイヌと異なることが示唆され、とくに低塩素化OH-PCBsの毒性リスクは高いことが予想された。低塩素化OH-PCBsは血中でTH輸送タンパクとの競合結合や、THの硫酸抱合排泄の阻害、TH起因性遺伝子の転写抑制などが報告されており、ネコの甲状腺機能障害が懸念される。
• 犬の膵炎診断における客観的で簡便な指標による超音波検査の有用性の検討

  竹内 恭介, 中村 健介, 森下 啓太郎, 大田 寛, 佐々木 東, 滝口 満喜  日本獣医師会雑誌  69-  (5)  292  -292  2016/05  [Not refereed][Not invited]
  
• 右心機能の低下は僧帽弁疾患犬の生存期間を短縮する

  森田 智也, 中村 健介, 大菅 辰幸, 佐々木 東, 大田 寛, 滝口 満喜  日本獣医師会雑誌  69-  (5)  294  -294  2016/05  [Not refereed][Not invited]
  
• 日本の専門家が結集して作る獣医臨床腫瘍学テキストブック 消化器系(食道〜結腸・直腸、肛門) 概要

  滝口 満喜  Veterinary Oncology  3-  (2)  78  -84  2016/04
• シスプララチンデリバリーの効果が及ぶ距離

  佐々木東, 工藤信樹, 中村健介, 森下啓太郎, 大田寛, 滝口満喜  超音波医学  43-  (Suppl.)  S544  -S544  2016/04  [Not refereed][Not invited]
  
• 有機ハロゲン化合物によるコンパニオンアニマルの汚染実態と甲状腺ホルモンへの影響評価

  野見山桂, 山本恭央, 水川葉月, 江口哲史, 滝口満喜, 中山翔太, 池中良徳, 石塚真由美, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  25th-  2016
• 本当にソノポレーションか?

  佐々木東, 工藤信樹, 中山翔太, 滝口満喜  日本超音波医学会光超音波画像研究会プログラム・抄録集  2016-  (2nd)  22‐24  2016  [Not refereed][Not invited]
  
• 犬の急性および慢性膵炎の診断治療の最前線 膵炎の画像診断と生検診断

  滝口 満喜  動物臨床医学  24-  (4)  153  -154  2015/12
• 右心室‐左心室‐左心房interaction:犬急性肺高血圧症モデルにおける検討

  大菅辰幸, 中村健介, 森田智也, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  46-  (Suppl.2)  251  -251  2015/12  [Not refereed][Not invited]
  
• 診察中に左房破裂による心タンポナーデを発症したものの内科療法のみで長期生存中の犬の1例

  川元誠, 中村健介, 高瀬奈美, 森田智也, 大菅辰幸, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  46-  (Suppl.2)  241  -241  2015/12  [Not refereed][Not invited]
  
• 二次診療施設において診断・治療に苦慮した消化器疾患の症例カンファレンス 膵臓疾患編

  滝口 満喜  動物臨床医学会年次大会プロシーディング  36回-  (1)  125  -125  2015/11
• 胆管原発カルチノイドによる肝外胆管閉塞と診断した猫の1例

  森俊輔, 中村健介, 高木哲, 賀川由美子, 森下啓太郎, 大田寛, 滝口満喜  動物臨床医学会年次大会プロシーディング  36回-  (2)  283  -284  2015/11  [Not refereed][Not invited]
  
• 犬の非再生性免疫介在性貧血(NRIMA)に対する治療の前向き検討

  森下啓太郎, 竹内恭介, 森俊輔, 早川小百合, 中村健介, 佐々木東, 大田寛, 滝口満喜  動物臨床医学会年次大会プロシーディング  36回-  (2)  367  -368  2015/11  [Not refereed][Not invited]
  
• 犬のびまん性肝疾患に対する造影超音波検査の応用の試み

  道下 明日香, 森下 啓太郎, 華園 究, 中村 健介, 滝口 満喜, 高木 哲  日本獣医画像診断学会抄録  58回-  12  -12  2015/09
• ミニチュア・ダックスフンドの炎症性結直腸ポリープ症例の結直腸粘膜におけるToll‐like receptor2,4の局在解析

  横山望, 大田寛, 賀川由美子, 山崎淳平, 市居修, KHOIRUN Nisa, 森田智也, 大菅辰幸, 佐々木東, 森下啓太郎, 中村健介, 滝口満喜  日本獣医学会学術集会講演要旨集  158th-  402  -402  2015/08/30  [Not refereed][Not invited]
  
• 胸腺腫に続発した後天性重症筋無力症の猫の1例

  早川小百合, 大田寛, 細谷謙次, 賀川由美子, 華園究, 田淵祐司, 滝口満喜  北海道獣医師会雑誌  59-  (8)  403  -403  2015/08/10  [Not refereed][Not invited]
  
• 【一目でわかる症候シリーズvol.4 嘔吐(1)】胃の腫瘍

  滝口 満喜  SA Medicine  17-  (4)  18  -21  2015/08
• (パート1)参加型臨床実習をどうするか 目前にせまった参加型臨床実習の内容、実施方法を議論する 伴侶動物の参加型臨床実習 国立大学編国立大学の参加型臨床実習の実施方針、実際の実施計画(伴侶動物)「国際認証を目指す伴侶動物の参加型臨床実習」

  滝口 満喜  日本獣医学会学術集会講演要旨集  158回-  139  -139  2015/08
• 汎用性が高く臨床的にも有用な犬の左心房機能評価法の検討;僧帽弁逆流の重症度評価について

  川元誠, 中村健介, 大菅辰幸, 森田智也, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  北海道獣医師会雑誌  59-  (8)  390  -390  2015/08  [Not refereed][Not invited]
  
• 右心機能の低下は僧帽弁疾患犬の生存期間を短縮する

  森田智也, 中村健介, 大菅辰幸, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  59-  (8)  389  -389  2015/08  [Not refereed][Not invited]
  
• 超低脂肪食で治療した犬慢性腸症18例の回顧的検討

  横山望, 大田寛, 佐々木東, 森下啓太郎, 中村健介, 滝口満喜  北海道獣医師会雑誌  59-  (8)  394  -394  2015/08  [Not refereed][Not invited]
  
• 犬の膵炎診断における客観的で簡便な指標による超音波検査の有用性の検討

  竹内恭介, 中村健介, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  北海道獣医師会雑誌  59-  (8)  396  -396  2015/08  [Not refereed][Not invited]
  
• 心エコー図検査による右心機能指標と心臓カテーテル検査との関連:急性肺高血圧モデル犬を用いた検討

  森田智也, 中村健介, 大菅辰幸, 森下啓太郎, 佐々木東, 大田寛, 滝口満喜  日本獣医学会学術集会講演要旨集  158回-  403  -403  2015/08  [Not refereed][Not invited]
  
• 免疫抑制治療中に合併したRgizopus microsporus感染に伴う胃穿孔の犬の1例

  那須 香菜子, 中村 健介, 高木 哲, 賀川 由美子, 青島 圭佑, 甲斐 千暁, 後藤 正光, 滝口 満喜  北海道獣医師会雑誌  59-  (8)  392  -392  2015/08  [Not refereed][Not invited]
  
• 【副腎腫瘍】副腎腫瘍の内科

  滝口 満喜  Joncol: Japanese Journal of Veterinary Clinical Oncology  11-  (2)  22  -24  2015/07
• Dynamic CTによる肝細胞性腫瘤の良悪性鑑別

  那須香菜子, 中村健介, 高木哲, 細谷謙次, 華園究, 賀川由美子, 森下啓太郎, 滝口満喜  日本獣医麻酔外科学雑誌  46-  (Supplement 1)  302  -302  2015/06/10  [Not refereed][Not invited]
  
• 右室Tei‐indexは僧帽弁疾患犬の1年生存を規定する

  森田智也, 中村健介, 大菅辰幸, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  46-  (Suppl.1)  280  -280  2015/06  [Not refereed][Not invited]
  
• 僧帽弁疾患犬におけるうっ血性心不全の発症はLA strain法によるブースターポンプ機能指標と関連する

  川元誠, 中村健介, 大菅辰幸, 森田智也, 佐々木東, 森下啓太郎, 大田寛, 滝口満喜  日本獣医麻酔外科学雑誌  46-  (Suppl.1)  281  -281  2015/06  [Not refereed][Not invited]
  
• 肝静脈造影超音波検査による肝疾患罹患犬の肝内血行動態の評価

  森下 啓太郎, 平本 彰, 大菅 辰幸, 中村 健介, 大田 寛, 滝口 満喜  日本獣医師会雑誌  68-  (5)  326  -327  2015/05  [Not refereed][Not invited]
  
• ドラッグデリバリーを可視化する培養系の開発

  佐々木 東, 滝口 満喜, Bos Clemens, Moonen Chrit  超音波医学  42-  (Suppl.)  S476  -S476  2015/04
• ドラッグデリバリーを可視化する培養系の開発

  佐々木 東, 滝口 満喜, Bos Clemens, Moonen Chrit  超音波医学  42-  (Suppl.)  S476  -S476  2015/04  [Not refereed][Not invited]
  
• 肝静脈造影超音波検査による肝疾患罹患犬の肝内血行動態の評価

  森下啓太郎, 平本彰, 大菅辰幸, 中村健介, 大田寛, 滝口満喜  日本獣医師会獣医学術学会年次大会講演要旨集  2014-  137  2015/01/30  [Not refereed][Not invited]
  
• ペットネコにおける有機ハロゲン化合物および水酸化代謝物の汚染実態と甲状腺ホルモン恒常性への影響

  山本恭央, 野見山桂, 水川葉月, 滝口満喜, 石塚真由美, 国末達也, 田辺信介  環境化学討論会要旨集(CD-ROM)  24th-  2015
• ペット動物における有機ハロゲン化合物および水酸化代謝物の曝露実態と甲状腺ホルモン恒常性へ及ぼす影響

  山本恭央, 野見山桂, 水川葉月, 滝口満喜, 石塚真由美, 国末達也, 田辺信介  日本内分泌かく乱化学物質学会研究発表会要旨集  18th-  124  -124  2015
• 犬におけるSpeckle tracking法を用いた右室strain解析の再現性

  MORITA TOMOYA, NAKAMURA KENSUKE, OSUGA TATSUYUKI, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  45-  (Suppl.2)  216  -216  2014/12  [Not refereed][Not invited]
  
• 犬の急性および慢性膵炎の診断治療の最前線―膵炎の画像診断と生検診断―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  35th-  (1)  209  -210  2014/11/14  [Not refereed][Not invited]
  
• うっ血性左心不全を呈した原発性心臓血管肉腫の犬の1例

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, TOOYAMA NOBUO, MORITA TOMOYA, KAGAWA YUMIKO, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  35回-  (2)  33  -34  2014/11  [Not refereed][Not invited]
  
• 免疫抑制量のプレドニゾロンが奏功した重度肺高血圧症を伴う間質性肺疾患を疑ったウェルシュコーギーの1例

  MORITA TOMOYA, NAKAMURA KENSUKE, OSUGA TATSUYUKI, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  35回-  (2)  103  -104  2014/11  [Not refereed][Not invited]
  
• NSAIDsが原因と考えられた骨髄壊死の犬の1例

  MORISHITA KEITARO, DEGUCHI TATSUYA, KAGAWA YUMIKO, NAKAMURA KENSUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  35回-  (2)  93  -94  2014/11  [Not refereed][Not invited]
  
• 犬慢性消化器疾患における十二指腸造影超音波検査の有用性の検討

  篠原 正義, リン・スーイー, 森下 啓太郎, 横山 望, 大菅 辰幸, 中村 健介, 賀川 由美子, 大田 寛, 滝口 満喜  日本獣医画像診断学会抄録  56回-  24  -24  2014/09  [Not refereed][Not invited]
  
• 参加型臨床実習 小動物

  TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  157th-  192  -192  2014/08/11  [Not refereed][Not invited]
  
• 参加型臨床実習 小動物

  TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  398  -398  2014/08/10  [Not refereed][Not invited]
  
• プロカルバジンが奏功した肉芽腫性髄膜脳脊髄炎を疑う犬の1例

  YAMAGUCHI TOMOO, OTA HIROSHI, MATSUMOTO MASANORI, MORISHITA KEITARO, NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  365  -365  2014/08  [Not refereed][Not invited]
  
• 肺高血圧症の犬に対するDual pulsed‐wave Doppler法を用いた右室Tei‐index測定の有用性

  MORITA TOMOYA, NAKAMURA KENSUKE, OSUGA TATSUYUKI, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  369  -369  2014/08  [Not refereed][Not invited]
  
• 尿管結石による水腎症から膿腎症に至った犬の1例

  HAYAKAWA SAYURI, NAKAMURA KENSUKE, TAKAGI SATOSHI, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  376  -376  2014/08  [Not refereed][Not invited]
  
• 二次的な巨大食道を伴った鼻咽頭ポリープの猫の1例

  NASU KANAKO, NAKAMURA KENSUKE, TAKAGI SATOSHI, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  374  -374  2014/08  [Not refereed][Not invited]
  
• 活性型ビタミンD₃外用薬の慢性摂取により高カルシウム血症が認められた犬の2例

  SHIMOIDE ARISA, NAKAMURA KENSUKE, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  379  -379  2014/08  [Not refereed][Not invited]
  
• 肝静脈造影超音波検査による肝疾患罹患犬の肝内血行動態の評価

  MORISHITA KEITARO, HIRAMOTO AKIRA, RIN SU・I, OSUGA TATSUYUKI, NAKAMURA KENSUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  58-  (8)  370  -370  2014/08  [Not refereed][Not invited]
  
• 左心房機能は心血管薬によりいかに変化するか:正常犬における2D Speckle Tracking法を用いた検討

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, MORITA TOMOYA, YEE LIM SUE, YOKOYAMA NOZOMU, MORISHITA KEITARO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  157回-  479  -479  2014/08  [Not refereed][Not invited]
  
• ミニチュア・ダックスフンドの炎症性結直腸ポリープ症例の結直腸粘膜におけるclaudinの蛋白質発現解析

  YOKOYAMA NOZOMU, OTA HIROSHI, MORITA TOMOYA, OSUGA TATSUYUKI, LIM SUE YEE, MORISHITA KEITARO, NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  157回-  491  -491  2014/08  [Not refereed][Not invited]
  
• 犬バベハア症治療における熱ショック蛋白質90の薬剤標的としての可能性

  ABE MOEKO, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  157回-  361  -361  2014/08  [Not refereed][Not invited]
  
• 犬の非感染性髄膜脳炎症例の脳脊髄液中におけるサイトカインの発現解析

  OTA HIROSHI, KAMADA MAYUMI, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医神経病学会  40th-  19  2014/07/05  [Not refereed][Not invited]
  
• 犬胃癌10症例における超音波検査所見

  YOKOYAMA NOZOMI, OTA HIROSHI, NAKAMURA KENSUKE, MORISHITA KEITARO, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  45-  (Suppl.1)  261  -261  2014/06  [Not refereed][Not invited]
  
• 造影超音波検査による腎機能評価の基礎的検討

  MORISHITA KEITARO, RIN SU・I, OSUGA TATSUYUKI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  45-  (Suppl.1)  262  -262  2014/06  [Not refereed][Not invited]
  
• 徐脈性不整脈に対してピモベンダンを投与し失神の改善を認めた犬の1例

  MORITA TOMOYA, SUZUKI SHUJI, NAKAMURA KENSUKE, OSUGA TATSUYUKI, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  45-  (Suppl.1)  248  -248  2014/06  [Not refereed][Not invited]
  
• 2D Speckle Tracking法を用いた左心房機能評価による犬心筋症の重症度評価

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, SUZUKI SHUJI, MORISHITA KEITARO, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医師会雑誌  67-  (5)  367  -367  2014/05/20  [Not refereed][Not invited]
  
• プレドニゾロンを主とする免疫抑制療法実施症例における犬膵特異的リパーゼの推移

  OTA HIROSHI, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医師会雑誌  67-  (5)  369  -369  2014/05  [Not refereed][Not invited]
  
• セルレイン誘発膵炎犬における造影超音波検査の定量的および定性的評価

  LIM SUE YEE, NAKAMURA KENSUKE, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医畜産新報  67-  (4)  286  -288  2014/04  [Not refereed][Not invited]
   

  本研究ではコレシストキニン類似体セルレインによる急性膵炎モデル犬を使用して、急性膵炎の診断における造影超音波検査の有用性を検討することを目的とした。その結果、造影超音波検査は微小血管の変化を検出することが可能であり、これにより膵臓における炎症の存在を検出できる可能性が示された。特に、セルレイン誘発膵炎においては持続した強い造影増強が認められることが特徴的な所見であることが示された。(著者抄録)
• 北海道での小動物臨床での放射線利用の広がり

  TAKIGUCHI MITSUYOSHI  日本アイソトープ協会放射線安全取扱部会年次大会要旨集  2014-  57  2014  [Not refereed][Not invited]
  
• LC-MS/MS Analysis of Canine Lipoproteins Fractionated Using the Ultracentrifugation-Precipitation Method (vol 75, pg 1471, 2013)

  Asuka Suto, Masahiro Yamasaki, Yukari Takasaki, Yasuyuki Fujita, Riichiro Abe, Hiroshi Shimizu, Hiroshi Ohta, Mitsuyoshi Takiguchi  JOURNAL OF VETERINARY MEDICAL SCIENCE  75-  (12)  1678  -1678  2013/12  [Not refereed][Not invited]
  
• Dual pulsed‐wave doppler法による右室Tei‐index測定の再現性

  MORITA TOMOYA, NAKAMURA KENSUKE, OSUGA TATSUYUKI, SUZUKI SHUJI, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  44-  (Suppl.2)  250  -250  2013/12  [Not refereed][Not invited]
  
• 猫心筋症において2D Speckle Tracking法を用いた左心房機能評価は可能であるのか

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, SUZUKI SHUJI, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  44-  (Suppl.2)  249  -249  2013/12  [Not refereed][Not invited]
  
• 両側性尿管閉塞を呈した犬の1例

  TOOYAMA NOBUO, NAKAMURA KENSUKE, KAGAWA YUMIKO, MORISHITA KEITARO, OTA HIROSHI, TAKAGI SATOSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  34回-  (2)  3  -4  2013/11  [Not refereed][Not invited]
  
• 非典型的溶血性尿毒症症候群(D‐HUS)が疑われた犬の1例

  MORISHITA KEITARO, TOOYAMA NOBUO, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  34回-  (2)  141  -142  2013/11  [Not refereed][Not invited]
  
• 慢性特発性偽性腸閉塞症が疑われた犬の1例

  YOKOYAMA NOZOMU, OTA HIROSHI, HASHIBA KANO, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OCHIAI KENJI, TERAMOTO HIDESHI, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  34回-  (2)  59  -60  2013/11  [Not refereed][Not invited]
  
• 市販フードの給餌によって病的骨折を起こした栄養性二次性上皮小体機能亢進症の犬の1例

  HASHIBA KANO, MORISHITA KEITARO, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  34回-  (2)  119  -120  2013/11  [Not refereed][Not invited]
  
• 組織奇形が疑われた膀胱粘膜下嚢胞の犬の1例

  MORISHITA KEITARO, HOSOYA KENJI, OSUGA TATSUYUKI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  34回-  (2)  9  -10  2013/11  [Not refereed][Not invited]
  
• 超音波検査により診断が可能であった子宮捻転の犬の1例

  中村 健介, 山崎 真大, 大田 寛, 森下 啓太郎, 木村 享史, 滝口 満喜  日本獣医画像診断学会抄録  54回-  28  -28  2013/09  [Not refereed][Not invited]
  
• 胆嚢十二指腸吻合および胆嚢肝管吻合を行った胆管閉塞の猫の1症例

  ANEGAWA HIROFUMI, TAKAGI SATOSHI, HASHIBA KANO, HOSOYA KENJI, HOSHINO YUKI, TAKIGUCHI MITSUYOSHI, OKUMURA MASAHIRO  北海道獣医師会雑誌  57-  (8)  400  -400  2013/08/10  [Not refereed][Not invited]
  
• プレドニゾロンを主とする免疫抑制療法実施症例における犬膵特異的リパーゼの推移

  OTA HIROSHI, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  57-  (8)  402  -402  2013/08  [Not refereed][Not invited]
  
• 輪状咽頭アカラシアの犬の1例

  UENO TAKASHI, OTA HIROSHI, TAKAGI SATOSHI, HOSHI KIYOTAKA, MORISHITA KEITARO, NAKAMURA TAKESHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  57-  (8)  401  -401  2013/08  [Not refereed][Not invited]
  
• 2D Speckle Tracking法を用いた左心房機能評価による犬心筋症の重症度評価

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, HOSOYA KENJI, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  57-  (8)  396  -396  2013/08  [Not refereed][Not invited]
  
• 非典型的溶血性尿毒症症候群(D‐HUS)が疑われた犬の1例

  TOOYAMA NOBUO, MORISHITA KEITARO, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  57-  (8)  395  -395  2013/08  [Not refereed][Not invited]
  
• 閉鎖式持続吸引ドレーンにて良好にコントロールできた膵膿瘍の犬の1例

  YAMAGUCHI TOMOO, MORISHITA KEITARO, HOSOYA KENJI, MUKAIDANI RYUSUKE, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  57-  (8)  400  -400  2013/08  [Not refereed][Not invited]
  
• ジミナゼン耐性Babesia gibsoniの熱ショック蛋白質90遺伝子発現量の測定

  IDAKA NATSUKI, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  156回-  236  -236  2013/08  [Not refereed][Not invited]
  
• 特発性収縮性心膜炎の犬の1例

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, HOSOYA KENJI, MORITA TOMOYA, KAGAWA YUMIKO, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  44-  (Suppl.1)  245  -245  2013/06  [Not refereed][Not invited]
  
• QUALITATIVE AND QUANTITATIVE CONTRAST ENHANCED ULTRASONOGRAPHIC ASSESSMENT OF CERULEIN-INDUCED PANCREATITIS IN DOGS

  S. Y. Lim, K. Nakamura, K. Morishita, H. Ohta, M. Yamasaki, M. Takiguchi  JOURNAL OF VETERINARY INTERNAL MEDICINE  27-  (3)  707  -707  2013/05  [Not refereed][Not invited]
  
• MARKED INCREASED EXPRESSION OF INTERLEUKIN 8 IN THE COLORECTAL MUCOSA OF DOGS WITH INFLAMMATORY COLORECTAL POLYPS BUT NOT WITH LYMPHOCYTIC-PLASMACYTIC COLITIS

  Y. Tamura, H. Ohta, N. Yokoyama, K. Morishita, K. Nakamura, M. Yamasaki, M. Takiguchi  JOURNAL OF VETERINARY INTERNAL MEDICINE  27-  (3)  706  -706  2013/05  [Not refereed][Not invited]
  
• モバイル超音波時代の幕開け

  TAKIGUCHI MITSUYOSHI  MVM  22-  (4)  61  -63  2013/05/01  [Not refereed][Not invited]
  
• 肝静脈血流波形解析による右心不全の重症度評価

  NAKAMURA KENSUKE, OSUGA TATSUYUKI, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医師会雑誌  66-  (5)  346  -347  2013/05  [Not refereed][Not invited]
  
• 内科領域の取り組みと課題

  TSUJIMOTO HAJIME, OKUDA MASARU, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  155th-  150  -150  2013/03/04  [Not refereed][Not invited]
  
• ミニチュア・ダックスフンドの炎症性結直腸ポリープの病態

  OTA HIROSHI, TAMURA YU, YOKOYAMA NOZOMI, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  155回-  153  -153  2013/03  [Not refereed][Not invited]
  
• Babesia gibsoni熱ショック蛋白質90の遺伝子クローニングおよび系統樹解析

  YAMASAKI MASAHIRO, TSUBOI YOSHIHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本寄生虫学会大会プログラム・抄録集  82nd-  84  2013/02  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第12回 膵臓を極める

  TAKIGUCHI MITSUYOSHI  SA Med  15-  (1)  18  -22  2013/02/01  [Not refereed][Not invited]
  
• 開業獣医師のための画像診断再入門 実例から学ぶ画像診断ピットフォール

  TAKIGUCHI MITSUYOSHI  MVM  22-  (1)  31  -36  2013/01/01  [Not refereed][Not invited]
  
• ―免疫抑制剤の理論と実践―炎症性腸疾患の概要と免疫抑制剤使用の考え方

  TAKIGUCHI MITSUYOSHI  CAP (Companion Anim Pract)  27-  (282)  19  -23  2012/12/01  [Not refereed][Not invited]
  
• 末梢肺動脈狭窄症と診断した高齢猫の1例

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  43-  (Suppl.2)  213  -213  2012/12  [Not refereed][Not invited]
  
• 最近よく診る消化器疾患の診断と治療―IBD,胃運動性障害等の対策について―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  33rd-  (3)  223  -225  2012/11/16  [Not refereed][Not invited]
  
• Babesia gibsoniにおけるdiminazene aceturate耐性の性質の安定性

  B.K.・ウィクラマセカラ・ラジャパクシャゲ, 山崎 真大, S.J.・ファン, 中村 健介, 佐々木 東, 村上 正紘, 田村 悠, S.Y.・リン, 大田 寛, 滝口 満喜  獣医寄生虫学会誌  11-  (1)  35  -35  2012/10  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第11回 内分泌疾患を超音波で診る(2)甲状腺・上皮小体

  TAKIGUCHI MITSUYOSHI  SA Med  14-  (5)  81  -87  2012/10/01  [Not refereed][Not invited]
  
• ソナゾイド造影超音波検査と機械学習を組み合わせた脾臓腫瘤の自動良悪性鑑別の試み

  中村 健介, 滝口 満喜, 高木 一也, 近藤 敏志  日本獣医画像診断学会抄録  52回-  16  -16  2012/09
• 北海道の飼育犬におけるCapnocytophaga属菌の侵淫調査

  HATANAKA YUI, TAKIGUCHI MITSUYOSHI, GOTO MASAMITSU, MURAMATSU YASUKAZU  日本獣医学会学術集会講演要旨集  154th-  270  -270  2012/08/31  [Not refereed][Not invited]
  
• 披裂軟骨背側に形成された炎症性腫瘤病変を喉頭切開により摘出した猫の1例

  IWANO KIMI, TAKAGI SATOSHI, HOSHINO YUKI, HOSOYA KENJI, TAKIGUCHI MITSUYOSHI, OKUMURA MASAHIRO  北海道獣医師会雑誌  56-  (8)  411  -411  2012/08/10  [Not refereed][Not invited]
  
• 好酸球増多症候群を伴った消化器型リンパ腫の猫の1例

  TOOYAMA NOBUO, HOSOYA KENJI, NAKAMURA KENSUKE, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  56-  (8)  399  -399  2012/08  [Not refereed][Not invited]
  
• 下垂体性副腎皮質機能亢進症とクロム親和性細胞腫を併発した犬の1例

  KANAZAWA TOMONORI, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, TAKAGI SATOSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  56-  (8)  410  -410  2012/08  [Not refereed][Not invited]
  
• 麻酔後の全脳虚血が疑われた2症例のMRI所見

  OTA HIROSHI, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  56-  (8)  429  -429  2012/08  [Not refereed][Not invited]
  
• 肝静脈血流波形解析による右心不全の重症度評価

  NAKAMURA KENSUKE, OSUGA TATSUYUKI, MORISHITA KEITARO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  56-  (8)  409  -409  2012/08  [Not refereed][Not invited]
  
• 両側性子宮分節性無形成から片側性子宮角捻転に至った犬の1例

  YOKOYAMA NOZOMI, NAKAMURA KENSUKE, KIMURA TAKASHI, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  56-  (8)  418  -418  2012/08  [Not refereed][Not invited]
  
• 下垂体性副腎皮質機能亢進症と中枢性尿崩症を併発し,放射線治療を行った下垂体腫瘍の犬の1例

  TAMURA YU, YAMASAKI MASAHIRO, MORISHITA KEITARO, NAKAMURA KENSUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  56-  (8)  410  -410  2012/08  [Not refereed][Not invited]
  
• Diminazene aceturate耐性Babesia gibsoniの核酸取り込み量の測定

  WATANABE NATSUO, YAMASAKI MASAHIRO, KUMARA W, R. BANDULA, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  154回-  215  -215  2012/08  [Not refereed][Not invited]
  
• ミニチュア・ダックスフンドの炎症性結直腸ポリープ病変における炎症性サイトカインの発現解析

  TAMURA YU, OTA HIROSHI, MURAKAMI MASAHIRO, BANDULA KUMARA WR, SUE YEE LIM, OSUGA TATSUYUKI, MORISHITA KEITARO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  154回-  307  -307  2012/08  [Not refereed][Not invited]
  
• 腎細胞癌移植マウスにおけるマルチキナーゼ阻害剤Sorafenib治療後のFLTによる治療反応評価

  MURAKAMI MASAHIRO, CHO SHOKICHI, CHO GEN, CHODORI NUSURATTOFATIMA, CHODORI NUSURATTOFATIMA, U BUMBUN, NISHIJIMA KEN'ICHI, KUBO NAOKI, TAKIGUCHI MITSUYOSHI, TAMAKI NAGARA, KUGE YUJI  核医学  49-  (3)  S246  -S246  2012/08  [Not refereed][Not invited]
  
• 犬の頭蓋内疾患診断における経頭蓋超音波ドプラ法の有用性の検討

  SASAOKA KAZUYOSHI, NAKAMURA KENSUKE, MORISHITA KEITARO, OSUGA TATSUYUKI, LIM SUE YEE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  43-  (Suppl.1)  270  -270  2012/06  [Not refereed][Not invited]
  
• CT検査を実施した肝葉捻転の犬の2例

  MORISHITA KEITARO, HOSOYA KENJI, TAKAGI SATOSHI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  43-  (Suppl.1)  267  -267  2012/06  [Not refereed][Not invited]
  
• 房室ブロックの犬2例に認められた低速僧帽弁逆流と高速左室駆出血流

  NAKAMURA KENSUKE, TOOYAMA NOBUO, UENO TAKASHI, MORISHITA KEITARO, OSUGA TATSUYUKI, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  43-  (Suppl.1)  245  -245  2012/06  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第10回 内分泌疾患を超音波で診る(1)副腎を極める

  TAKIGUCHI MITSUYOSHI  SA Med  14-  (3)  68  -73  2012/06/01  [Not refereed][Not invited]
  
• LEFT ATRIAL FUNCTION ANALYSIS BY LEFT ATRIAL TRACKING METHOD IN CANINE CHRONIC MITRAL VALVULAR HEART DISEASE.

  K. Nakamura, T. Osuga, M. Yamasaki, H. Ohta, M. Takiguchi  JOURNAL OF VETERINARY INTERNAL MEDICINE  26-  (3)  720  -720  2012/05  [Not refereed][Not invited]
  
• INTERLEUKIN-17A GENE EXPRESSION IN THE INTESTINAL MUCOSA IN DOGS WITH LYMPHOCYTIC-PLASMACYTIC ENTERITIS AND INFLAMMATORY COLORECTAL POLYP

  H. Ohta, K. Takada, Y. Tamura, K. Nakamura, M. Yamasaki, M. Takiguchi  JOURNAL OF VETERINARY INTERNAL MEDICINE  26-  (3)  772  -773  2012/05  [Not refereed][Not invited]
  
• ¹⁸F×FMISOを用いた腎細胞癌移植モデルにおける抗血管新生療法(sorafenicの早期治療反応の評価

  MURAKAMI MASAHIRO, CHO GEN, NUSURATTOFATIMA CHODORI, U MOMBUN, NISHIJIMA KEN'ICHI, TAKIGUCHI MITSUYOSHI, TAMAKI NAGARA, KUGE YUJI  JSMI Rep  5-  (2)  117  -117  2012/05  [Not refereed][Not invited]
  
• パルス超音波を用いたソノポレーションの実現に向けたいくつかの基礎的検討

  KUDO NOBUKI, SASAKI NOBORU, TAKIGUCHI MITSUYOSHI  超音波医学  39-  (Suppl.)  S174  -S174  2012/04/15  [Not refereed][Not invited]
  
• 診断装置によるソノポレーションの特徴と条件設定

  SASAKI NOBORU, NAKAMURA KENSUKE, KUDO NOBUKI, TAKIGUCHI MITSUYOSHI  超音波医学  39-  (2)  226  -227  2012/03/15  [Not refereed][Not invited]
  
• ジミナゼン耐性株および野生株におけるBabesia gibsoniミトコンドリアDNAの塩基配列の比較

  ウイクラマセカラ B.K., ラジャパク・シャゲ, 山崎 真大, 中村 健介, 佐々木 東, 村上 正紘, 田村 悠, リン S.Y., 大田 寛, 滝口 満喜  獣医寄生虫学会誌  10-  (1-2)  34  -34  2012/03  [Not refereed][Not invited]
  
• Babesia gibsoniにおけるdiminazene aceturate抵抗性の安定性(Stability of characteristic of the diminazene aceturate resistance in Babesia gibsoni)

  Rajapakshage Bandula Kumara Wickramasekara, 山崎 真大, Hwang Shiang-Jyi, 中村 健介, 佐々木 東, 村上 正紘, 田村 悠, Lim Sue Yee, 大田 寛, 滝口 満喜  日本獣医学会学術集会講演要旨集  153回-  221  -221  2012/03  [Not refereed][Not invited]
  
• 臨床の立場から臨床薬理学に期待するもの

  TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  153rd-  147  -147  2012/03/01  [Not refereed][Not invited]
  
• 診断シリーズ vol.1 肝胆道系疾患 X線検査

  TAKIGUCHI MITSUYOSHI  SA Med  14-  (1)  37  -39  2012/02/01  [Not refereed][Not invited]
  
• 診断シリーズ vol.1 肝胆道系疾患 超音波検査

  TAKIGUCHI MITSUYOSHI  SA Med  14-  (1)  40  -44  2012/02/01  [Not refereed][Not invited]
  
• ここまでは押さえておこう! 腹部エコー検査

  滝口 満喜  日本獣医画像診断学会抄録  50回-  59  -59  2012/01
• 麻酔後の全脳虚血が疑われた猫2例のMRI所見

  大田 寛, 山崎 真大, 滝口 満喜  日本獣医画像診断学会抄録  50回-  94  -94  2012/01  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第9回 嘔吐の原因を超音波で探る(4)腸管を極める その2

  TAKIGUCHI MITSUYOSHI  SA Med  13-  (6)  55  -58  2011/12/01  [Not refereed][Not invited]
  
• 心エコー図検査にて心内膜心筋線維症と診断した猫4例

  NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, MURAKAMI MASAHIRO, MATSUYAMA ARATA, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  42-  (Suppl.2)  233  -233  2011/12  [Not refereed][Not invited]
  
• 活性型ビタミンD₃外用薬(マキサカルシトール軟膏)常用患者の飼育犬に認められた高カルシウム血症

  TOOYAMA NOBUO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  32nd-  (2)  149  -150  2011/11/18  [Not refereed][Not invited]
  
• 最近よく診る消化器疾患の傾向と対策―確実な診断と治療を行うには―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  32nd-  (3)  185  -185  2011/11/18  [Not refereed][Not invited]
  
• 内分泌疾患の画像診断

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  32nd-  (1)  100  -100  2011/11/18  [Not refereed][Not invited]
  
• 小型犬における下垂体性副腎皮質機能亢進症の超音波診断基準

  MATSUYAMA ARATA, TOOYAMA NOBUO, MURAKAMI MASAHIRO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  32nd-  (3)  14  -18  2011/11/18  [Not refereed][Not invited]
  
• ポリエン系抗生物質アムホテリシンBのイヌバベシア症治療薬としての応用に関する研究

  YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  32nd-  (3)  91  -94  2011/11/18  [Not refereed][Not invited]
  
• 超音波診断装置および超音波造影剤による抗がん剤の抗腫瘍効果の増強

  SASAKI NOBORU, NAKAMURA KENSUKE, KUDO NOBUKI, TAKIGUCHI MITSUYOSHI  超音波医学  38-  (6)  674  -674  2011/11/15  [Not refereed][Not invited]
  
• 最近よく診る消化器疾患の診断と治療

  TAKIGUCHI MITSUYOSHI  MVM  20-  (7)  82  -87  2011/11/01  [Not refereed][Not invited]
  
• 犬の肝臓腫瘤診断におけるソナゾイド造影超音波検査―4年間の検討―

  NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, HOSOYA KENJI, TAKAGI SATOSHI, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  32回-  (3)  6  -9  2011/11  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第8回 嘔吐の原因を超音波で探る(3)腸管を極める その1

  TAKIGUCHI MITSUYOSHI  SA Med  13-  (5)  79  -83  2011/10/01  [Not refereed][Not invited]
  
• セフォベシンが有効であった巨大食道症に併発した誤嚥性肺炎の犬の一例

  TAKIGUCHI MITSUYOSHI  CAP (Companion Anim Pract)  26-  (268)  78  -80  2011/10/01  [Not refereed][Not invited]
  
• 犬の嘔吐の治療および管理の実際と対応 制吐剤の効果的な使い方

  滝口 満喜, 大野 耕一, 竹内 和義, 峯岸 則之, 米元 仁  CLINIC NOTE  7-  (9)  77  -83  2011/09
• 病理組織検査にて角層下および真皮への多数の好酸球の浸潤が認められた犬の2症例

  CHIBA ERI, YAMASAKI MASAHIRO, OTA HIROSHI, NAKAMURA KENSUKE, SASAKI NOBORU, MURAKAMI MASAHIRO, TAMURA YU, YEE LIM SUE, KUMARA WICKRAMASEKARA R.B, HONDA HIDETAKA, NASUNO TOYOHIKO, OCHIAI KENJI, TAKIGUCHI MITSUYOSHI  獣医臨床皮膚科  17-  (3)  177  -182  2011/09  [Not refereed][Not invited]
  
• ジミナゼンアセチュレート耐性Babesia gibsoni株と野生型におけるミトコンドリアDNA配列の比較(Comparison of the sequence of mitochondrial DNA from diminazene aceturate resistance Babesia gibsoni strain with those from wild type)

  Rajapakshage Bandula Kumara Wickramasekara, 山崎 真大, 中村 健介, 佐々木 東, 村上 正紘, 田村 悠, Lim Sue Yee, 大田 寛, 滝口 満喜  日本獣医学会学術集会講演要旨集  152回-  203  -203  2011/08  [Not refereed][Not invited]
  
• セフォベシンが有効であった好中球性胆管炎の猫の一例

  TAKIGUCHI MITSUYOSHI  CAP (Companion Anim Pract)  26-  (266)  84  -86  2011/08/01  [Not refereed][Not invited]
  
• ”診る目を養う”超音波検査 第7回 嘔吐の原因を超音波で探る(2)胃の超音波検査 その2

  TAKIGUCHI MITSUYOSHI  SA Med  13-  (4)  70  -73  2011/08/01  [Not refereed][Not invited]
  
• 犬の僧房弁閉鎖不全症におけるLeft atrial tracking法を用いた左心房機能評価の有用性

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  55-  (8)  386  -386  2011/08  [Not refereed][Not invited]
  
• 後鼻孔領域に炎症性ポリープを認めた若齢犬の1例

  WATANABE TAKESHI, OTA HIROSHI, SONO FUMIO, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  55-  (8)  387  -387  2011/08  [Not refereed][Not invited]
  
• 気腫性胆嚢炎の犬の1例

  TAMURA YU, YAMASAKI MASAHIRO, NAKAMURA KENSUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  55-  (8)  386  -386  2011/08  [Not refereed][Not invited]
  
• 胃癌の犬5例における超音波検査および内視鏡検査所見の特徴

  TOOYAMA NOBUO, OTA HIROSHI, TAKAGI SATOSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  55-  (8)  397  -397  2011/08  [Not refereed][Not invited]
  
• 心エコー図検査とD‐dimer測定により診断し得た肺血栓塞栓症の犬の1例

  OGI NAOKO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  55-  (8)  385  -385  2011/08  [Not refereed][Not invited]
  
• 超音波診断装置および超音波造影剤による抗がん剤のソノポレーションの抗腫瘍効果

  SASAKI NOBORU, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, BANDULA KUMARA WR, LIM SUE YEE, TAMURA YU, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  152回-  302  -302  2011/08  [Not refereed][Not invited]
  
• 犬の慢性腸炎症例の腸粘膜におけるIL‐17A遺伝子の発現解析

  TAKADA KANAE, OTA HIROSHI, NAKAMURA KENSUKE, SASAKI NOBORU, MURAKAMI MASAHIRO, KUMARA W.R. BANDULA, TAMURA YU, LIM SUE YEE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  152回-  298  -298  2011/08  [Not refereed][Not invited]
  
• プロテオーム解析を用いた犬リポ蛋白質測定法の評価

  SUDO ASUKA, YAMASAKI MASAHIRO, NAKAMURA KENSUKE, SASAKI NOBORU, MURAKAMI MASAHIRO, KUMARA W.R. BANDULA, TAMURA YU, LIM SUE YEE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  152回-  291  -291  2011/08  [Not refereed][Not invited]
  
• 犬の慢性腸炎症例の結腸粘膜における細胞接着分子クローディンの発現変動

  OTA HIROSHI, TAMURA YU, LIM SUE YEE, KUMARA WR BANDULA, MURAKAMI MASAHIRO, SASAKI NOBORU, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  152回-  298  -298  2011/08  [Not refereed][Not invited]
  
• 門脈体循環シャント(前編)門脈体循環シャントの画像診断:超音波検査

  TAKIGUCHI MITSUYOSHI, NAKAMURA KENSUKE  Surgeon  15-  (4)  33  -38  2011/07/01  [Not refereed][Not invited]
  
• ”診る目を養う”超音波検査 第6回 嘔吐の原因を超音波で探る(1)胃の超音波検査

  TAKIGUCHI MITSUYOSHI, NAKAMURA KENSUKE  SA Med  13-  (3)  72  -78  2011/06/01  [Not refereed][Not invited]
  
• 治療シリーズ Vol.10―私はこうしている―「肝胆道系・膵外分泌疾患」胆嚢炎

  TAKIGUCHI MITSUYOSHI  SA Med  13-  (3)  50  -51  2011/06/01  [Not refereed][Not invited]
  
• 犬の心疾患におけるLeft atrial tracking法を用いた左心房機能評価の有用性

  OSUGA TATSUYUKI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  42-  (Suppl.1)  243  -243  2011/06  [Not refereed][Not invited]
  
• 治療シリーズ Vol.10―私はこうしている―「肝胆道系・膵外分泌疾患」胆石症

  TAKIGUCHI MITSUYOSHI  SA Med  13-  (3)  51  -52  2011/06/01  [Not refereed][Not invited]
  
• 仮想気管支鏡検査が診断に有用であった肺葉捻転の犬の2例

  MURAKAMI MASAHIRO, NAKAMURA KENSUKE, SASAKI NOBORU, MATSUYAMA SHIN, TAMURA YU, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  42-  (Suppl.1)  264  -264  2011/06  [Not refereed][Not invited]
  
• 犬の肝カルチノイド4症例におけるソナゾイド造影超音波検査所見

  NAKAMURA KENSUKE, HOSOYA KENJI, YAMASAKI MASAHIRO, OTA HIROSHI, YEE LIM SUE, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  42-  (Suppl.1)  261  -261  2011/06  [Not refereed][Not invited]
  
• EFFECTS OF SILDENAFIL CITRATE ON DOGS WITH EISENMENGER'S SYNDROME

  K. Nakamura, M. Yamasaki, H. Ohta, M. Takiguchi  JOURNAL OF VETERINARY INTERNAL MEDICINE  25-  (3)  660  -660  2011/05  [Not refereed][Not invited]
  
• パルス超音波と微小気泡によるシスプラチンの抗腫瘍効果増強の基礎的知見

  SASAKI NOBORU, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, OTA HIROSHI, YAMASAKI MASAHIRO, KUDO NOBUKI, TAKIGUCHI MITSUYOSHI  超音波医学  38-  (3)  338  -338  2011/05  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第5回 門脈シャントを超音波で診断する

  NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  SA Med  13-  (2)  86  -90  2011/04/01  [Not refereed][Not invited]
  
• 診断用超音波とマイクロバブルによるドラッグデリバリーのin vitroでの最適化

  SASAKI NOBORU, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, KUMARA BADULA, OTA HIROSHI, YAMASAKI MASAHIRO, KUDO NOBUKI, TAKIGUCHI MITSUYOSHI  超音波医学  38-  (Suppl.)  S316  -S316  2011/04  [Not refereed][Not invited]
  
• シトシンアラビノシドにより良好に維持されている特発性脳炎の犬の2例

  OTA HIROSHI, SASAKI NOBORU, MURAKAMI MASAHIRO, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医皮膚科学会学術大会・総会  14th-  (Vol.1)  117  2011/03/11  [Not refereed][Not invited]
  
• 二次性赤血球増加症を伴ったEisenmenger症候群の犬に対するシルデナフィルの有効性の検討

  NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医皮膚科学会学術大会・総会  14th-  (Vol.1)  175  2011/03/11  [Not refereed][Not invited]
  
• 内分泌疾患の画像診断

  TAKIGUCHI MITSUYOSHI  日本獣医皮膚科学会学術大会・総会  14th-  (Vol.2)  272  2011/03/11  [Not refereed][Not invited]
  
• 超音波とマイクロバブルによるドラッグデリバリーに薬剤濃度とバブル濃度が及ぼす影響

  SASAKI NOBORU, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, KUMARA BANDULA, TAMURA YU, YEE LIM SUE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医皮膚科学会学術大会・総会  14th-  (Vol.1)  183  2011/03/11  [Not refereed][Not invited]
  
• アムホテリシンBによるイヌバベシア症の治療に関する研究

  YAMASAKI MASAHIRO, O KOJAKU, BANDULA KUMARA W. R, TANIYAMA YUSUKE, TSUBOI YOSHIHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  151回-  216  -216  2011/03  [Not refereed][Not invited]
  
• 甲状腺の外科(外科適応となる)甲状腺疾患の病態と診断

  TAKIGUCHI MITSUYOSHI  Surgeon  15-  (2)  8  -17  2011/03/01  [Not refereed][Not invited]
  
• 第二世代超音波造影剤ソナゾイドを用いた造影超音波検査による犬の肝臓および脾臓腫瘤の診断に関する研究

  NAKAMURA KENSUKE, TAKAGI SATOSHI, YAMASAKI MASAHIRO, OTA HIROSHI, SASAKI NOBORU, MURAKAMI MASAHIRO, TAMURA YU, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  151回-  82  -82  2011/03  [Not refereed][Not invited]
  
• アムホテリシンBのin vivoにおける抗バベシア効果に関する研究

  YAMASAKI MASAHIRO, HWANG SHIANG-JYI, KUMARA W.R.BANDULA, TANIYAMA YUSUKE, TSUBOI YOSHIHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本寄生虫学会大会プログラム・抄録集  80th-  73  2011/02  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第4回 肝臓腫瘤を超音波で鑑別する

  NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  SA Med  13-  (1)  101  -107  2011/02/01  [Not refereed][Not invited]
  
• シトシンアラビノシドが奏効した肉芽腫性髄膜脳脊髄炎疑いの犬の1例

  OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  55-  (1)  9  -11  2011/01  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第3回 黄疸を超音波で鑑別する(2)肝実質の超音波検査

  NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  SA Med  12-  (6)  65  -69  2010/12/01  [Not refereed][Not invited]
  
• 急性膵炎とどう戦う―急性膵炎の病態生理と診断―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  31st-  (3)  152  -153  2010/11/19  [Not refereed][Not invited]
  
• 腹部エコーのブラッシュ・アップ―上腹部を極める―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  31st-  (1)  1  -1  2010/11/19  [Not refereed][Not invited]
  
• 消化器疾患の内科学的アプローチ―診断技術の向上と的確な治療を目指して―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  31st-  (3)  187  -187  2010/11/19  [Not refereed][Not invited]
  
• ガストリン産生性カルチノイドに対し徐放性オクトレオチド製剤を使用した犬の1例

  細谷 謙次, 金子 一成, 滝口 満喜, 中村 健介, 高木 哲, 奥村 正裕  動物臨床医学会年次大会プロシーディング  31回-  (2)  193  -194  2010/11  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第2回 黄疸を超音波で鑑別する(1)胆道系の超音波検査

  TAKIGUCHI MITSUYOSHI, NAKAMURA KENSUKE  SA Med  12-  (5)  79  -85  2010/10/01  [Not refereed][Not invited]
  
• 教育の質保証と獣医学モデル・コア・カリキュラムおよび獣医学共用試験の役割 「eラーニングコンテンツ作成の試み」内科学

  滝口 満喜  日本獣医学会学術集会講演要旨集  150回-  131  -131  2010/09
• 微小気泡存在下での診断用超音波照射によるシスプラチンの抗腫瘍効果増強の検討

  SASAKI AZUMA, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, BANDULA KUMARA, TAMURA YU, OTA HIROSHI, YAMASAKI MASAHIRO, KUDO NOBUKI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  150回-  297  -297  2010/09  [Not refereed][Not invited]
  
• 犬のガストリノーマに対してサンドスタチンLARを投与した1例

  金子 一成, 細谷 謙次, 滝口 満喜, 中村 健介, 北村 二郎, 高木 哲, 奥村 正裕  北海道獣医師会雑誌  54-  (8)  450  -450  2010/08  [Not refereed][Not invited]
  
• “診る目を養う”超音波検査 第1回 腹部エコー:基本と上達のコツ

  TAKIGUCHI MITSUYOSHI  SA Med  12-  (4)  67  -71  2010/08/01  [Not refereed][Not invited]
  
• 造影超音波検査により病変の特定が可能であったインスリノーマの犬2例

  NAKAMURA KENSUKE, TAKAGI SATOSHI, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  460  -460  2010/08  [Not refereed][Not invited]
  
• 胆嚢粘液嚢腫の犬19症例の回顧的解析

  MURATA NAOMITSU, TAKAGI SATOSHI, HOSOYA KENJI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI, SATO KOTA, OKUMURA MASAHIRO  北海道獣医師会雑誌  54-  (8)  462  -462  2010/08  [Not refereed][Not invited]
  
• 細菌性心内膜炎の犬の1例

  MORI SHINSUKE, OTA HIROSHI, SAKAI HIDEFUMI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  459  -459  2010/08  [Not refereed][Not invited]
  
• 犬の皮膚無菌性肉芽腫/化膿性肉芽腫症候群と診断した1症例

  WAKAKI NOZOMI, YAMASAKI MASAHIRO, OTA HIROSHI, NASUNO TOYOHIKO, OCHIAI KENJI, TAKEBAYASHI KENSAKU, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  464  -464  2010/08  [Not refereed][Not invited]
  
• リンパ管拡張症を疑う胸水貯留に続発して肺葉捻転を起こした犬の1例

  MURAKAMI MASAHIRO, NAKAMURA KENSUKE, SASAKI AZUMA, OTA HIROSHI, YAMASAKI MASAHIRO, TAKAGI SATOSHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  464  -464  2010/08  [Not refereed][Not invited]
  
• 造影3DCTにより確定診断し得た犬の重複大動脈弓の1例

  MATSUYAMA SHIN, KANEKO KAZUNARI, YAMANE YOSHIHISA, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  458  -458  2010/08  [Not refereed][Not invited]
  
• 心房中隔欠損症と僧帽弁閉鎖不全症を併発した犬の1例

  OGI NAOKO, NAKAMURA KENSUKE, MATSUYAMA SHIN, HASHIBA MINORU, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  458  -458  2010/08  [Not refereed][Not invited]
  
• 経皮的エコーガイド下胆嚢穿刺により確定診断した細菌性胆嚢炎の犬と猫の2例

  UKAI AYAKO, KANEKO KAZUNARI, OKAWA MINAKO, OTA HIROSHI, KOBAYASHI HIDEAKI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  54-  (8)  461  -461  2010/08  [Not refereed][Not invited]
  
• 二次性赤血球増加症を伴ったEisenmenger症候群の犬に対するシルデナフィルの有効性の検討

  NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  41-  (Suppl.1)  294  -294  2010/07  [Not refereed][Not invited]
  
• CONTRAST-ENHANCED ULTRASONOGRAPHY WITH SONAZOID (R) FOR CHARACTERIZATION OF FOCAL SPLENIC LESIONS

  K. Nakamura, H. Ohta, M. Yamasaki, M. Takiguchi  JOURNAL OF VETERINARY INTERNAL MEDICINE  24-  (3)  733  -734  2010/05  [Not refereed][Not invited]
  
• 培養Babesia gibsoniの末梢血単核球への侵入と増殖の検討

  TANIYAMA YUSUKE, YAMASAKI MASAHIRO, HWANG SHIANG-JYI, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本寄生虫学会大会プログラム・抄録集  79th-  70  2010/05  [Not refereed][Not invited]
  
• ビジュアルで学ぶ!内視鏡検査と治療 第7回 手技編 下部消化管の内視鏡検査

  TAKIGUCHI MITSUYOSHI  MVM  19-  (4)  68  -69  2010/05/01  [Not refereed][Not invited]
  
• 超音波診断装置と微小気泡によるシスプラチンの抗腫瘍効果の増強

  SASAKI NOBORU, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, KUMARA BANDULA, OTA HIROSHI, YAMASAKI MASAHIRO, KUDO NOBUKI, TAKIGUCHI MITSUYOSHI  超音波医学  37-  (Suppl.)  S322  -S322  2010/04  [Not refereed][Not invited]
  
• Diminazene aceturate耐性Babesia gibsoni株に対する他の抗バベシア原虫薬の効果

  黄 香寂, 谷山 裕介, 山崎 真大, 大田 寛, 滝口 満喜  獣医寄生虫学会誌  8-  (2)  149  -149  2010/03  [Not refereed][Not invited]
  
• イヌおよびネコの腎疾患における病態差―腎機能とスリット膜関連分子の動態―

  ICHII OSAMU, YABUKI AKIRA, SASAKI NOBUYA, OTA HIROSHI, TAKIGUCHI MITSUYOSHI, OTSUKA SAORI, HASHIMOTO YOSHIHARU, ENDO DAIJI, KON YASUHIRO  日本獣医学会学術集会講演要旨集  149回-  213  -213  2010/03  [Not refereed][Not invited]
  
• 犬の肝臓腫瘤30例に対するソナゾイド造影超音波検査

  NAKAMURA KENSUKE, TAKAGI SATOSHI, YAMASAKI MASAHIRO, OTA HIROSHI, SASAKI NOBORU, TAKIGUCHI MITSUYOSHI  日本獣医師会三学会年次大会講演要旨集  2009-  237  2010/01/07  [Not refereed][Not invited]
  
• 診断と内科治療

  TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  40-  (Supplement 2)  99  -99  2009/12/10  [Not refereed][Not invited]
  
• 外科手術を実施した犬の肝細胞癌17症例の検討

  TAKAGI SATOSHI, TAKIGUCHI MITSUYOSHI, YAMASAKI MASAHIRO, OTA HIROSHI, NAKAMURA KENSUKE, OSAKI TOMOHIRO, HOSOYA YOSHITSUGU, OKUMURA MASAHIRO  獣医麻酔外科学雑誌  40-  (Suppl.2)  196  -196  2009/12  [Not refereed][Not invited]
  
• 犬の消化器疾患の内科学的アプローチ―最新の診断技術と治療法の紹介―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  30th-  (3)  247  -247  2009/11/20  [Not refereed][Not invited]
  
• 腹部エコー:基本と上達への道

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  30th-  (1)  9  -9  2009/11/20  [Not refereed][Not invited]
  
• 犬のリンパ球プラズマ細胞性腸炎における細胞接着分子クローディンの発現変動

  OTA HIROSHI, MURAKAMI MASAHIRO, KUMARA BANDURA, SASAKI NOBORU, NAKAMURA KENSUKE, KO KOJAKU, YAMAZAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  148回-  261  -261  2009/09  [Not refereed][Not invited]
  
• Diminazene aceturate耐性Babesia gibsoni株に対する他の抗バベシア原虫薬の効果

  KO KOJAKU, TANIYAMA YUSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  148回-  188  -188  2009/09  [Not refereed][Not invited]
  
• 犬の脾臓腫瘤におけるソナゾイド造影超音波検査の有用性

  NAKAMURA KENSUKE, YAMASAKI MASAHIRO, OTA HIROSHI, SASAKI NOBORU, MURAKAMI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  148回-  268  -268  2009/09  [Not refereed][Not invited]
  
• 犬の肝臓腫瘤28結節に対するソナゾイド造影超音波検査

  NAKAMURA KENSUKE, TAKAGI SATOSHI, SASAKI AKIRA, MURAKAMI MASAHIRO, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  53-  (8)  101  -101  2009/08  [Not refereed][Not invited]
  
• 胆嚢粘液嚢腫の犬5例における周術期管理

  MITSUDA KENTA, TAKAGI SATOSHI, SUNAGA TAKAFUMI, TAKIGUCHI MITSUYOSHI, OTA HIROSHI, SATO KOTA, OSAKI TOMOHIRO, OKUMURA MASAHIRO  北海道獣医師会雑誌  53-  (8)  93  -93  2009/08  [Not refereed][Not invited]
  
• 大脳辺縁系の壊死が疑われた猫の1症例

  KAMIJO HIROMI, OTA HIROSHI, MATSUMOTO MASANORI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  53-  (8)  103  -103  2009/08  [Not refereed][Not invited]
  
• 筋炎の併発が疑われた劇症型重症筋無力症の犬の1例

  UKAI AYAKO, YOSHIKAWA MUNEYOSHI, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, NAKAO KIICHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  53-  (8)  98  -98  2009/08  [Not refereed][Not invited]
  
• 消化器系の診療手技 消化管の超音波検査

  TAKIGUCHI MITSUYOSHI  SA Med  11-  (4)  24  -33  2009/08/01  [Not refereed][Not invited]
  
• 本態性高Na血症が疑われた神経症状を示した犬と猫の2例

  MURAKAMI MASAHIRO, OTA HIROSHI, NAKAMURA KENSUKE, SASAKI AKIRA, YAMASAKI MASAHIRO, ETO MASAYUKI, KOSEKI YUMI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  53-  (8)  97  -97  2009/08  [Not refereed][Not invited]
  
• 気管支内視鏡検査で気管支狭窄を認めた慢性発咳の犬の1症例

  SASAKI AKIRA, OTA HIROSHI, ANDO REMI, NAKAMURA KENSUKE, MURAKAMI MASAHIRO, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  53-  (8)  88  -88  2009/08  [Not refereed][Not invited]
  
• ボストンテリアの脾臓捻転の1症例

  OTA HIROSHI, TAKAGI SATOSHI, NAKAZAWA SHIGEKI, MURAKAMI MASAHIRO, SASAKI AKIRA, NAKAMURA KENSUKE, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  53-  (8)  89  -89  2009/08  [Not refereed][Not invited]
  
• 腹部エコーの基礎とブラッシュ・アップ

  TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  40-  (Supplement 1)  150  -150  2009/06/20  [Not refereed][Not invited]
  
• 腎疾患の新しい診断指標―Interleukin 1 family,member 6は尿細管傷害マーカーになり得る―

  ICHII OSAMU, KON YASUHIRO, SASAKI NOBUYA, HASHIMOTO YOSHIHARU, OTA HIROSHI, TAKIGUCHI MITSUYOSHI, YABUKI AKIRA, ENDO DAIJI  日本獣医学会学術集会講演要旨集  147回-  79  -79  2009/03  [Not refereed][Not invited]
  
• 培養Babesia gibsoniのストレス環境下におけるheat shock protein 70の役割に関する研究

  YAMASAKI MASAHIRO, HWANG SHIANG-JYI, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本寄生虫学会大会プログラム・抄録集  78th-  86  2009/02  [Not refereed][Not invited]
  
• SLEが疑われた血球貪食症候群の猫の1例

  YOSHIKAWA MUNEYOSHI, NAKAMURA KENSUKE, SASAKI AZUMA, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医師会三学会年次大会講演要旨集  2008-  296  2008/12/25  [Not refereed][Not invited]
  
• ポリエン系抗生物質アムホテリシンBの抗Babesia gibsoni効果

  原田 絵梨子, 山崎 真大, 黄 寂香, 大田 寛, 滝口 満喜  獣医寄生虫学会誌  7-  (2)  90  -90  2008/12  [Not refereed][Not invited]
  
• Diminazene aceturate耐性Babesia gibsoni株の作成

  黄 香寂, 山崎 真大, 大田 寛, 滝口 満喜  獣医寄生虫学会誌  7-  (2)  92  -92  2008/12  [Not refereed][Not invited]
  
• 初心者のための基本的な消化管の画像診断(X線,超音波,内視鏡)

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  29th-  (1)  110  -110  2008/11/14  [Not refereed][Not invited]
  
• 簡便な犬リポ蛋白質の測定法の開発と臨床例への応用

  TAKASAKI YUKARI, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  29回-  (3)  37  -40  2008/11  [Not refereed][Not invited]
  
• 超音波検査により求めた正常なビーグル犬の甲状腺体積と体重の相関

  SASAKI NOBORU, NAKAMURA KENSUKE, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  29回-  (3)  29  -32  2008/11  [Not refereed][Not invited]
  
• 犬の肝臓における造影超音波検査の有用性の検討

  NAKAMURA KENSUKE, SASAKI HIGASHI, YOSHIKAWA MUNEYOSHI, O KOJAKU, OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  146回-  265  -265  2008/09  [Not refereed][Not invited]
  
• 犬の消化菅粘膜におけるクローディン分子群の発現:十二指腸を中心として

  YAMAGUCHI TOMOO, OTA HIROSHI, NAKAMURA KENSUKE, O KOJAKU, SASAKI HIGASHI, YOSHIKAWA MUNEYOSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  146回-  258  -258  2008/09  [Not refereed][Not invited]
  
• Diminazene aceturate耐性Babesia gibsoni株の作成

  O KOJAKU, YAMASAKI MASAHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  146回-  173  -173  2008/09  [Not refereed][Not invited]
  
• ポリエン系抗生物質アムホテリシンBの抗Babesia gibsoni効果

  HARADA ERIKO, YAMASAKI MASAHIRO, O KOJAKU, OTA HIROSHI, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  146回-  172  -172  2008/09  [Not refereed][Not invited]
  
• 抗リン脂質抗体症候群が疑われた不明熱のミニチュア・ダックスフンドの1例

  NAKAMURA KENSUKE, SASAKI NOBORU, YOSHIKAWA MUNEYOSHI, OTA HIROSHI, YAMAZAKI MASAHIRO, OBA MASAYO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  52-  (8)  410  -410  2008/08  [Not refereed][Not invited]
  
• 犬の無菌性好酸球性膿疱症と診断した2症例

  MINAMIYAMA ERI, YAMAZAKI MASAHIRO, OTA HIROSHI, HONDA HIDETAKA, NASUNO TOYOHIKO, OCHIAI KENJI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  52-  (8)  413  -413  2008/08  [Not refereed][Not invited]
  
• 気胸を併発した先天性肺葉性肺気腫の犬の1例

  KAMIJO HIROMI, TAKAGI SATOSHI, OTA HIROSHI, YAMAZAKI MASAHIRO, KIMURA TAKASHI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  52-  (8)  420  -420  2008/08  [Not refereed][Not invited]
  
• エリスロポエチン依存性の二次性多血症が疑われた腎リンパ腫の犬の1例

  SASAKI AZUMA, YOSHIKAWA MUNEYOSHI, NAKAMURA KENSUKE, OTA HIROSHI, YAMAZAKI MASAHIRO, SATO KOTA, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  52-  (8)  415  -415  2008/08  [Not refereed][Not invited]
  
• SLEが疑われた血球貪食症候群の猫の1例

  YOSHIKAWA MUNEYOSHI, SHODA MASATO, NAKAMURA KENSUKE, SASAKI NOBORU, OTA HIROSHI, YAMAZAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  52-  (8)  410  -410  2008/08  [Not refereed][Not invited]
  
• 犬の炎症性腸疾患:リンパ球形質細胞性腸炎・結腸炎を中心に

  OTA HIROSHI, YAMASAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  52-  (7)  263  -265  2008/07  [Not refereed][Not invited]
  
• 第二世代超音波造影剤ソナゾイドによる肝臓腫瘤の診断

  NAKAMURA KENSUKE, OTA HIROSHI, YAMAZAKI MASAHIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  39-  (Suppl.1)  227  -227  2008/06  [Not refereed][Not invited]
  
• 腹部エコー検査のススメ―小動物臨床における超音波造影剤の可能性

  NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  CAP (Companion Anim Pract)  23-  (225)  23  -28  2008/03/01  [Not refereed][Not invited]
  
• クエン酸シルデナフィルが奏功した逆行性動脈管開存症のイヌの2例

  NAKAMURA KENSUKE, MORISHITA KEITARO, NAKADE TETSUYA, TAKIGUCHI MITSUYOSHI  日本獣医師会三学会年次大会講演要旨集  2007-  245  2008  [Not refereed][Not invited]
  
• 第二世代超音波造影剤ソナゾイドの小動物臨床への応用に関する基礎的検討

  TAKIGUCHI MITSUYOSHI, NAKAMURA KENSUKE  獣医麻酔外科学雑誌  38-  (Supplement 2)  290  -291  2007/11/20  [Not refereed][Not invited]
  
• 犬の膵疾患における超音波ヒストグラム解析の有用性に関する基礎的研究

  NAKAMURA KENSUKE, MORISHITA KEITARO, TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  144th-  160  -160  2007/08/27  [Not refereed][Not invited]
  
• クエン酸シルデナフィルが奏功した逆行性動脈管開存症の犬の2例

  NAKAMURA KENSUKE, MORISHITA KEITARO, NAKAIDE TETSUYA, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  51-  (8)  333  -333  2007/08/01  [Not refereed][Not invited]
  
• メソトレキセートが奏功したステロイド抵抗性炎症性腸疾患の犬の1例

  MORISHITA KEITARO, NAKAMURA KENSUKE, HIRAYAMA KAZUKO, TANIYAMA HIROYUKI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  51-  (8)  335  -335  2007/08/01  [Not refereed][Not invited]
  
• デジタルX線(DR)撮影下における非選択的血管造影法により逆行性動脈管開存症と診断された犬の2例

  MORISHITA KEITARO, NAKAMURA KENSUKE, NAKAIDE TETSUYA, TAKIGUCHI MITSUYOSHI  日本獣医循環器学会抄録  86th-  (Suppl.1)  203  -203  2007  [Not refereed][Not invited]
  
• 十二指腸に閉塞性病変を伴った炎症性陽疾患の犬の1例

  中村 健介, 森下 啓太郎, 廉澤 剛, 平山 和子, 谷山 弘行, 滝口 満喜  北海道獣医師会雑誌  50-  (8)  79  -79  2006/08
• I型偽性低アルドステロン症が疑われた犬の1例

  MORISHITA KEITARO, NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  50-  (8)  336  -80  2006/08/01  [Not refereed][Not invited]
  
• 犬の慢性リンパ性白血病:メルファラン継続投与による良好な長期コントロールの1例

  HIROSE TAKESHI, NAKAMURA KENSUKE, MORISHITA KEITARO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  50-  (8)  346  -90  2006/08/01  [Not refereed][Not invited]
  
• デジタルX線(DR)撮影下における非選択的血管造影法により逆行性動脈管開存症と診断された犬の1例

  TOOYAMA NOBUO, NAKAMURA KENSUKE, MORISHITA KEITARO, NAKAIDE TETSUYA, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  50-  (8)  355  -99  2006/08/01  [Not refereed][Not invited]
  
• 十二指腸に閉塞性病変を伴った炎症性腸疾患の犬の1例

  NAKAMURA KENSUKE, MORISHITA KEITARO, RENZAWA TSUYOSHI, HIRAYAMA KAZUKO, TANIYAMA HIROYUKI, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  50-  (8)  335  2006/08/01  [Not refereed][Not invited]
  
• 肉芽腫性胃炎により幽門狭窄症を呈したイヌの一例

  NAKAMURA KENSUKE, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  37-  (Supplement 1)  207  -207  2006/06/20  [Not refereed][Not invited]
  
• 壊死性髄膜脳炎(NME)が強く疑われた犬8例と抗アストロサイト抗体

  NAKADE TETSUYA, MATSUKI NAOAKI, INOUE AIKA, TAKIGUCHI MITSUYOSHI, NIIYAMA MASAYOSHI, IZUMISAWA YASUHARU, TANADA ATSUSHI, YOSHIOKA YU, YOSHIOKA HINATSU, WATANABE TAKUO, TERASAWA ASAKO, ENOMOTO KOICHI, ISHIZUKA KISEKO, TAMAI SATOSHI  動物臨床医学会年次大会プロシーディング  26th-  (2)  195  -196  2005/11/18  [Not refereed][Not invited]
  
• 臨床的に線維軟骨性塞栓症と診断されたイヌのMRI所見

  MIYOSHI KENJIRO, NAKADE TETSUYA, IZUMISAWA YASUHARU, TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  26th-  (3)  26  -29  2005/11/18  [Not refereed][Not invited]
  
• 食道狭窄:とくに猫,バルーンなどを使った治療

  TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  36-  (Supplement 2)  73  -73  2005/10/01  [Not refereed][Not invited]
  
• 私立および国立獣医大学における臨床講座の違いとそれぞれの教育・研究のあり方

  TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  140th-  30  -30  2005/08/31  [Not refereed][Not invited]
  
• 犬の明らかな血小板増多を伴った赤芽球癆

  町田 竜彦, 内田 英二, 伊藤 真之, 木村 真, 滝口 満喜, 谷山 弘行  北海道獣医師会雑誌  49-  (8)  71  -71  2005/08
• 眼底出血から副腎腫瘍が疑われた犬の1例

  若生 晋輔, 滝口 満喜, 前原 誠也, 久保 明, 都築 圭子, 伊藤 典彦, 泉澤 康晴  北海道獣医師会雑誌  49-  (8)  73  -73  2005/08
• 犬猫の消化管内異物の超音波診断

  TAKIGUCHI MITSUYOSHI  獣医画像診断  (16/17)  3  -8  2005/08  [Not refereed][Not invited]
  
• 眼底出血から副腎腫ようが疑われた犬の1例

  WAKO SHINSUKE, TAKIGUCHI MITSUYOSHI, MAEHARA MASAYA, KUBO AKIRA, TSUZUKI KEIKO, ITO NORIHIKO, IZUMISAWA YASUHARU  北海道獣医師会雑誌  49-  (8)  289  2005/08/01  [Not refereed][Not invited]
  
• 頻回の内視鏡下バルーン拡張術が功を奏した食道狭窄の猫の1例

  MIWA ERIKO, MIYATA KAYOKO, MACHIDA TATSUHIKO, NAKAMURA KENSUKE, NAKAIDE TETSUYA, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  49-  (8)  276  -60  2005/08/01  [Not refereed][Not invited]
  
• 犬の明らかな血小板増多を伴った赤芽球ろう

  MACHIDA TATSUHIKO, UCHIDA EIJI, ITO MASAYUKI, KIMURA MAKOTO, TAKIGUCHI MITSUYOSHI, TANIYAMA HIROYUKI  北海道獣医師会雑誌  49-  (8)  287  2005/08/01  [Not refereed][Not invited]
  
• 超音波検査により粥状動脈硬化症と診断された犬の1例

  NAKAMURA KENSUKE, KITAMURA JIRO, TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  49-  (8)  293  -77  2005/08/01  [Not refereed][Not invited]
  
• 血管超音波検査によりアテローム性動脈硬化症が疑われた犬の一例

  中村 健介, 北村 二郎, 滝口 満喜  日本獣医循環器学会抄録  82-  249  -249  2005/06/25
• 最近の腹部超音波診断の有効性

  TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  36-  (Supplement 1)  120  -120  2005/06/20  [Not refereed][Not invited]
  
• 血管超音波検査によりアテローム性動脈硬化症が疑われた犬の一例

  NAKAMURA KENSUKE, KITAMURA JIRO, TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  36-  (Supplement 1)  240  -240  2005/06/20  [Not refereed][Not invited]
  
• Autoimmunity against a tissue kallikrein in IQI/Jic mice - A model for Sjogren's syndrome

  K Takada, M Takiguchi, A Konno, A Inaba  JOURNAL OF BIOLOGICAL CHEMISTRY  280-  (5)  3982  -3988  2005/02  [Not refereed][Not invited]
   

  We have recently characterized IQI/Jic mice as a model for Sjogren's syndrome (SS), a chronic autoimmune disease in humans. In SS, local lymphocytic infiltrations into salivary and lacrimal glands frequently develop to the involvement of systemic exocrine and nonexocrine organs, and the mechanism for progression of this disease remains obscure. Herein, we report identification of an autoantigen shared by various target organs in IQI/Jic mice. Polypeptides identified based on immunorecognition by autoantibodies in sera from IQI/ Jic mice affected with autoimmune disease (> 12 weeks of age) were tissue kallikrein (Klk)-1 and -13 and were cross-reactive to the autoantibodies. Interestingly, Klk-13, but not Klk-1, caused a proliferative response of splenic T cells from IQI/Jic mice from the age of 4 weeks onward. In addition, remarkably enhanced expression of Klk-13 was observed in the salivary glands of the mice in accordance with the development of inflammatory lesions. These results indicate that Klk-13 acts as an autoantigen and may increase T cells responsive to organs commonly expressing Klk-13, playing a pivotal role in the etiology of progression of disease in IQI/Jic mice. Our findings provide insights into the contributions of autoantigens shared by multiple organs in the progress of SS from an organ-specific to a systemic disorder.
• 紐状異物による十二指腸の穿孔に対して腸管内側から修復を行った犬の1例

  HOSHINO YUKI, KADOSAWA TSUYOSHI, TAKIGUCHI MITSUYOSHI, OKUMURA MASAHIRO, FUJINAGA TOORU  獣医麻酔外科学雑誌  36-  (1)  19  -22  2005/01/31  [Not refereed][Not invited]
  
• 犬のポリープ様ぼうこう炎の超音波画像所見

  TAKIGUCHI MITSUYOSHI  日本産業動物獣医学会・日本小動物獣医学会・日本獣医公衆衛生学会年次大会  2004-  295  2005  [Not refereed][Not invited]
  
• 胸部,腹部の超音波診断法

  TAKIGUCHI MITSUYOSHI  日本産業動物獣医学会・日本小動物獣医学会・日本獣医公衆衛生学会年次大会  2004-  212  -215  2005  [Not refereed][Not invited]
  
• 腎臓の障害をいかにして知るか―腎疾患の画像診断―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  25th-  (1)  326  -326  2004/11/19  [Not refereed][Not invited]
  
• 犬と猫の急性すい炎のすべてがわかるセミナー―急性すい炎の画像診断―

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  25th-  (3)  240  -241  2004/11/19  [Not refereed][Not invited]
  
• 脳溢血と診断した小型初老犬の1例

  天内 満雄, 滝口 満喜, 黒田 敏  北海道獣医師会雑誌  48-  (10)  4  -6  2004/10
• IVRによりコイル栓塞術を実施したイヌ動脈管開存症の1例

  NAKADE TETSUYA, RENZAWA TAKESHI, TAKIGUCHI MITSUYOSHI, ASANO KAZUYUKI  獣医麻酔外科学雑誌  35-  (Supplement 2)  61  -61  2004/10/01  [Not refereed][Not invited]
  
• 脳溢血と診断した小型初老犬の1例

  AMAUCHI MITSUO, TAKIGUCHI MITSUYOSHI, KURODA SATOSHI  北海道獣医師会雑誌  48-  (10)  422  -424  2004/10/01  [Not refereed][Not invited]
  
• リパーゼ産生性すい臓腫ようによる無菌性脂肪織炎が疑われた犬の1例

  SEKIGUCHI MICHIKO, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  北海道獣医師会雑誌  48-  (8)  305  -305  2004/08/01  [Not refereed][Not invited]
  
• ホジキン型リンパ腫が疑われた猫の1例

  OKAZAKI KAZUKO, TAKIGUCHI MITSUYOSHI, KIMURA TAKAFUMI, MATSUDA EIICHIRO, INABA MUTSUMI  北海道獣医師会雑誌  48-  (8)  319  -319  2004/08/01  [Not refereed][Not invited]
  
• 新生子牛のグロビンスイッチングにおける転写因子erythroid Kruppel‐like factorの役割

  ARASHIKI NOBUTO, ITO DAISUKE, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  138th-  148  -148  2004/08/01  [Not refereed][Not invited]
  
• 反すう動物赤血球膜タンパク質gp155の分子実態はMRP4である

  HIRAI MAYUMI, ITO DAISUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  138回-  178  -178  2004/08  [Not refereed][Not invited]
  
• クローディン‐16欠損胎子腎の組織構築とタイトジャンクション構成分子の発現動態

  OTA HIROSHI, ITO DAISUKE, KAGEYAMA SOICHI, HIRAYAMA HIROKI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  138回-  148  -148  2004/08  [Not refereed][Not invited]
  
• 血尿を主訴とした雌ウサギの子宮疾患例

  NAGANO MASASHI, KATAGIRI SEIJI, TAKIGUCHI MITSUYOSHI, KOMATSU TOMOHIKO, NAGASHIMA TARO, IKAWA SHIN'ICHI, TAKAHASHI YOSHIYUKI  北海道獣医師会雑誌  48-  (8)  318  -318  2004/08/01  [Not refereed][Not invited]
  
• 犬のポリープ様ぼうこう炎の超音波画像

  TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  北海道獣医師会雑誌  48-  (8)  307  -307  2004/08/01  [Not refereed][Not invited]
  
• シェーグレン症候群モデルマウス・IQI/Jicにおける自己抗原の同定

  TAKADA KENSUKE, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  138th-  160  -160  2004/08/01  [Not refereed][Not invited]
  
• バンド3の細胞膜移行におけるC末端細胞質領域の重要性

  ITO DAISUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  138回-  178  -178  2004/08  [Not refereed][Not invited]
  
• Spontaneous development of multiple glandular and extraglandular lesions in aged IQI/Jic mice: a model for primary Sjogren's syndrome

  K Takada, M Takiguchi, A Konno, M Inaba  RHEUMATOLOGY  43-  (7)  858  -862  2004/07  [Not refereed][Not invited]
   

  Objective. In primary Sjogren's syndrome (SS), systemic exocrine and non-exocrine organs are frequently affected, in addition to the major target tissues of the lacrimal and salivary glands. This study aimed to examine whether the IQI/Jic mouse, an animal model of SS whose autoimmune dacryoadenitis and sialoadenitis have been documented, develops inflammatory lesions in multiple organs as in primary SS. Methods. Systemic histopathological analysis was performed on IQI/Jic mice at various ages. Phenotypes of infiltrated lymphocytes were determined using immunohistochemical techniques. Results. Inflammatory lesions were observed not only in the lacrimal and salivary glands, but also in multiple organs, including the lung, pancreas and kidney at advanced ages, and were mainly composed of CD4(+) T cells and B cells. The incidence and severity of the inflammatory lesions increased with age in all these organs. The histological appearance and spreading of lesions were similar to those in human primary SS. Conclusions. IQI/Jic mice spontaneously develop inflammatory cellular infiltrates in multiple exocrine and non-exocrine organs. This characteristic distinguishes IQI/Jic mice from other murine models, making them favourable for studies on the pathogenesis of systemic involvement in primary SS.
• Spontaneous development of multiple glandular and extraglandular lesions in aged IQI/Jic mice: a model for primary Sjogren's syndrome

  K Takada, M Takiguchi, A Konno, M Inaba  RHEUMATOLOGY  43-  (7)  858  -862  2004/07  [Not refereed][Not invited]
   

  Objective. In primary Sjogren's syndrome (SS), systemic exocrine and non-exocrine organs are frequently affected, in addition to the major target tissues of the lacrimal and salivary glands. This study aimed to examine whether the IQI/Jic mouse, an animal model of SS whose autoimmune dacryoadenitis and sialoadenitis have been documented, develops inflammatory lesions in multiple organs as in primary SS. Methods. Systemic histopathological analysis was performed on IQI/Jic mice at various ages. Phenotypes of infiltrated lymphocytes were determined using immunohistochemical techniques. Results. Inflammatory lesions were observed not only in the lacrimal and salivary glands, but also in multiple organs, including the lung, pancreas and kidney at advanced ages, and were mainly composed of CD4(+) T cells and B cells. The incidence and severity of the inflammatory lesions increased with age in all these organs. The histological appearance and spreading of lesions were similar to those in human primary SS. Conclusions. IQI/Jic mice spontaneously develop inflammatory cellular infiltrates in multiple exocrine and non-exocrine organs. This characteristic distinguishes IQI/Jic mice from other murine models, making them favourable for studies on the pathogenesis of systemic involvement in primary SS.
• 乳腺上皮細胞におけるタイトジャンクション構成タンパク質の発現動態

  OTA HIROSHI, KOMABAYASHI KEN'ICHI, ITO DAISUKE, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本膜学会年会講演要旨集  26th-  56  2004/04/30  [Not refereed][Not invited]
  
• 牛遺伝性バンド3欠損症ホモ接合型胎子における組織病態

  ITO DAISUKE, KAGEYAMA SOICHI, SAWAI KEN, NAGATA AKIHIRO, OTA HIROSHI, TAKIGUCHI MITSUYOSHI, BAN AKIRA, WATANABE DAISAKU, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  137回-  136  -136  2004/03  [Not refereed][Not invited]
  
• 乳腺管腔上皮におけるタイトジャンクション構成蛋白質の発現動態

  OTA HIROSHI, KOMABAYASHI KEN'ICHI, ITO DAISUKE, WATANABE ATSUSHI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  137回-  152  -152  2004/03  [Not refereed][Not invited]
  
• Autoimmunity against a tissue kallikrein in IQI/Jic mice, a model for Sjogren's syndrome 'jointly worked'

  J. Biol. Chem..  (in press)-  2004  [Not refereed][Not invited]
  
• Diagnostic ultrasound of polypoid cystitis in the dog 'jointly worked'

  J. Vet. Med. Sci.  (in press)-  2004  [Not refereed][Not invited]
  
• Diagnostic ultrasound of polypoid cystitis in the dog 'jointly worked'

  J. Vet. Med. Sci.  (in press)-  2004  [Not refereed][Not invited]
  
• 腎疾患の新たな診断と治療法を目指して 腎疾患の画像診断

  TAKIGUCHI MITSUYOSHI  獣医畜産新報  57-  (990)  35  -40  2004/01/01  [Not refereed][Not invited]
  
• Magnetic resonance imaging and immunoblot analyses in rats with experimentally induced cerebral alveolar echinococcosis

  T Asanuma, Y Matsumoto, M Takiguchi, O Inanami, M Nakao, K Nakaya, A Ito, A Hashimoto, M Kuwabara  COMPARATIVE MEDICINE  53-  (6)  649  -656  2003/12  [Not refereed][Not invited]
   

  The early stage of experimentally induced secondary cerebral alveolar echinococcosis (AE) in rats was investigated by use of magnetic resonance imaging (MRI) and immunoblot (western blot) analyses. Thirty-six female Wistar rats (6 to 8 weeks old) were injected intracranially with a 10% homogenate of echinococcal larval tissues in which the concentrations of microvesicles and protoscolices were estimated to be 3.8 and 1.5 x 10(4)/ml, respectively. To observe the fine structure of the rat brain, MRI was performed under a high magnetic field of 7.05 T. Histologic examination also was performed. The T2-weighted MR images revealed a hyperintense region in the cerebral cortex at two weeks after injection of the homogenate. At three weeks after injection, this region was found to have cysts on the basis of results of histologic examination. Signal-void regions corresponding to hyperplasia and the subsequent calcification of the cuticle layer at six and 13 weeks after injection, respectively, were observed in T2-weighted and proton density MR images. On the other hand, at nine weeks after injection, AE was discernible by use of western blot analysis of sera with antibodies of two epitopes (Em18 and Em16) of E. multilocularis. Using this secondary cerebral AE animal model, it was concluded that the MRI method was suitable for early detection of secondary cerebral AE.
• Magnetic resonance imaging and immunoblot analyses in rats with experimentally induced cerebral alveolar echinococcosis

  T Asanuma, Y Matsumoto, M Takiguchi, O Inanami, M Nakao, K Nakaya, A Ito, A Hashimoto, M Kuwabara  COMPARATIVE MEDICINE  53-  (6)  649  -656  2003/12  [Not refereed][Not invited]
   

  The early stage of experimentally induced secondary cerebral alveolar echinococcosis (AE) in rats was investigated by use of magnetic resonance imaging (MRI) and immunoblot (western blot) analyses. Thirty-six female Wistar rats (6 to 8 weeks old) were injected intracranially with a 10% homogenate of echinococcal larval tissues in which the concentrations of microvesicles and protoscolices were estimated to be 3.8 and 1.5 x 10(4)/ml, respectively. To observe the fine structure of the rat brain, MRI was performed under a high magnetic field of 7.05 T. Histologic examination also was performed. The T2-weighted MR images revealed a hyperintense region in the cerebral cortex at two weeks after injection of the homogenate. At three weeks after injection, this region was found to have cysts on the basis of results of histologic examination. Signal-void regions corresponding to hyperplasia and the subsequent calcification of the cuticle layer at six and 13 weeks after injection, respectively, were observed in T2-weighted and proton density MR images. On the other hand, at nine weeks after injection, AE was discernible by use of western blot analysis of sera with antibodies of two epitopes (Em18 and Em16) of E. multilocularis. Using this secondary cerebral AE animal model, it was concluded that the MRI method was suitable for early detection of secondary cerebral AE.
• Feline leptin: Immunogenic and biological activities of the recombinant protein, and its measurement by ELISA

  H Shibata, N Sasaki, T Honjoh, Ohishi, I, M Takiguchi, K Ishioka, M Ahmed, M Soliman, K Kimura, M Saito  JOURNAL OF VETERINARY MEDICAL SCIENCE  65-  (11)  1207  -1211  2003/11  [Not refereed][Not invited]
   

  Leptin is a protein synthesized and secreted primarily by adipose tissue. The blood leptin concentration is known to reflect body fat content in rodents, humans and dogs, and thereby is useful for quantitative assessment of obesity. In the present study, we produced recombinant feline leptin in Escherichia coli transfected with feline leptin cDNA we cloned previously. The recombinant feline leptin with a molecular weight of 16 kDa induced phosphorylation of the signal transducers and activators of transcription 3 (STAT3) protein in the cells expressing rat leptin receptor. The anti-feline leptin antibody raised in rabbits reacted well to feline and human leptin and less to rodents' leptin in Western blot analysis. Sandwich enzyme-linked immunosorbent assay (ELISA) was developed, using rabbit anti-feline leptin antibody and recombinant feline leptin as a standard. In this ELISA system, cross-reactivity to human, rat and mouse leptin was 30.7%, 69.5% and 66.6%, respectively. The plasma leptin levels of 24 healthy cats were in a range from 0.3 to 29.7 ng/ml with the mean +/- SEM of 4.5 +/- 1.3 ng/ml, being positively proportional to body fat content. These results indicate that our ELISA system may be useful for assessment of obesity in cats.
• Feline leptin: Immunogenic and biological activities of the recombinant protein, and its measurement by ELISA

  H Shibata, N Sasaki, T Honjoh, Ohishi, I, M Takiguchi, K Ishioka, M Ahmed, M Soliman, K Kimura, M Saito  JOURNAL OF VETERINARY MEDICAL SCIENCE  65-  (11)  1207  -1211  2003/11  [Not refereed][Not invited]
   

  Leptin is a protein synthesized and secreted primarily by adipose tissue. The blood leptin concentration is known to reflect body fat content in rodents, humans and dogs, and thereby is useful for quantitative assessment of obesity. In the present study, we produced recombinant feline leptin in Escherichia coli transfected with feline leptin cDNA we cloned previously. The recombinant feline leptin with a molecular weight of 16 kDa induced phosphorylation of the signal transducers and activators of transcription 3 (STAT3) protein in the cells expressing rat leptin receptor. The anti-feline leptin antibody raised in rabbits reacted well to feline and human leptin and less to rodents' leptin in Western blot analysis. Sandwich enzyme-linked immunosorbent assay (ELISA) was developed, using rabbit anti-feline leptin antibody and recombinant feline leptin as a standard. In this ELISA system, cross-reactivity to human, rat and mouse leptin was 30.7%, 69.5% and 66.6%, respectively. The plasma leptin levels of 24 healthy cats were in a range from 0.3 to 29.7 ng/ml with the mean +/- SEM of 4.5 +/- 1.3 ng/ml, being positively proportional to body fat content. These results indicate that our ELISA system may be useful for assessment of obesity in cats.
• 紐状異物による十二指腸の線状穿孔に対して腸管内側から修復を行った犬の一例

  HOSHINO YUKI, KADOSAWA TAKESHI, TAKIGUCHI MITSUYOSHI, MATSUHISA YUKI, OKUMURA MASAHIRO, FUJINAGA TOORU  獣医麻酔外科学雑誌  34-  (Supplement 2)  99  -100  2003/09/20  [Not refereed][Not invited]
  
• 調節帯異常により左室機能障害を呈した猫の超音波検査

  HAMAMOTO KOHEI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  136th-  205  -205  2003/09/10  [Not refereed][Not invited]
  
• シェーグレン症候群モデルマウス・IQI/Jicにおける多発性自己免疫病変の組織学的解析および自己抗原の同定

  TAKADA KENSUKE, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  136th-  218  -218  2003/09/10  [Not refereed][Not invited]
  
• 副腎性性ホルモン機能こう進症とポメラニアンの脱毛症X

  TAKIGUCHI MITSUYOSHI, TAKADA KENSUKE  日本獣医学会学術集会講演要旨集  136th-  54  -54  2003/09/10  [Not refereed][Not invited]
  
• 腎疾患の画像診断

  TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  136th-  50  -50  2003/09/10  [Not refereed][Not invited]
  
• 牛腎尿細管においてクローディン16と共存するパートナークローディンの解析と遺伝子の単離

  OTA HIROSHI, ITO DAISUKE, KOMABAYASHI KEN'ICHI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  136回-  196  -196  2003/09  [Not refereed][Not invited]
  
• バンド3欠損症変異バンド3認識・分解の分子機構―R664X変異周辺領域認識の検証―

  ITO DAISUKE, OTA HIROSHI, TAKIGUCHI MITSUYOSHI, INABA MUTSUMI  日本獣医学会学術集会講演要旨集  136回-  217  -217  2003/09  [Not refereed][Not invited]
  
• 右大動脈弓遺残と動脈管開存が併発した犬の1例

  KAWAMURA TAKAO, TAKIGUCHI MITSUYOSHI, HAMAMOTO KOHEI, HASEBE RIE, RENZAWA TAKESHI, INABA MUTSUMI  北海道獣医師会雑誌  47-  (8)  316  -316  2003/08/01  [Not refereed][Not invited]
  
• 甲状腺ホルモン製剤補充療法により病態が改善された拡張型心筋症の犬の1例

  OKAZAKI KAZUKO, TAKIGUCHI MITSUYOSHI, HAMAMOTO KOHEI, MURAOKA NOBUYOSHI, INABA ATSUSHI  北海道獣医師会雑誌  47-  (8)  295  -295  2003/08/01  [Not refereed][Not invited]
  
• 超音波検査によりしん出性収縮性心膜炎と診断された犬の1例

  HAMAMOTO KOHEI, TAKIGUCHI MITSUYOSHI, ITO AKIRA, KIMURA TAKASHI, RENZAWA TAKESHI, INABA MUTSUMI  北海道獣医師会雑誌  47-  (8)  315  -315  2003/08/01  [Not refereed][Not invited]
  
• 小動物の紐状異物の超音波画像診断

  TAKIGUCHI MITSUYOSHI, HAMAMOTO KOHEI, INABA MUTSUMI  北海道獣医師会雑誌  47-  (8)  317  -317  2003/08/01  [Not refereed][Not invited]
  
• 悪性組織球症が疑われた犬の2例

  ITO KEI, TAKIGUCHI MITSUYOSHI, HAMAMOTO KOHEI, NAKAO YOSHIKAZU, INABA ATSUSHI  北海道獣医師会雑誌  47-  (8)  291  -291  2003/08/01  [Not refereed][Not invited]
  
• 無菌性結節性脂肪織炎が疑われた犬の2例

  SEKIGUCHI MICHIKO, TAKIGUCHI MITSUYOSHI, INABA ATSUSHI  北海道獣医師会雑誌  47-  (8)  302  -302  2003/08/01  [Not refereed][Not invited]
  
• Presence of B7-2(+) dendritic cells and expression of Th1 cytokines in the early development of sialodacryoadenitis in the IQI/Jic mouse model of primary Sjogren's syndrome

  A Konno, K Takada, J Saegusa, M Takiguchi  AUTOIMMUNITY  36-  (4)  247  -254  2003/07  [Not refereed][Not invited]
   

  Subpopulations of infiltrating lymphocytes, professional antigen-presenting cells (APCs), and Th1/Th2 cytokines that could initiate an autoimmune sialodacryoadenitis were studied in the IQI/Jic mouse model of primary Sjogren's syndrome. Although lymphocytic infiltrations were first seen in submandibular glands (SMGs) of females and in lacrimal glands (LGs) of males at 8 weeks of age, clusters of MHC class II+ , CD11c(+) , B7-2(+) dendritic cells (DCs) were already localized in these tissues at 4 weeks. At 8 weeks, the infiltrating lymphocytes consisted of almost equal numbers of B cells and CD4(+) T cells. In the inflammatory foci, MHC class II+ , CD11c(+) , B7-2(+) DCs formed network-like structures. Duct cells in the lesions showed immunoreactivities for MHC class II and ALCAM (a costimulatory adhesion molecule). IL-12 and IFN-gamma transcripts were detected by RT-PCR in SMGs of females and in LGs of males at 8-12 weeks. These results suggest that the clustered DCs might play an important role in the initiation of the adenitis, and further suggest that the DCs and epithelial cells may participate in the activation of CD4(+) T cells. It is also likely that Th1 cytokines mediate the functional interactions between the APCs and CD4(+) T cells in the early lesions.
• Presence of B7-2(+) dendritic cells and expression of Th1 cytokines in the early development of sialodacryoadenitis in the IQI/Jic mouse model of primary Sjogren's syndrome

  A Konno, K Takada, J Saegusa, M Takiguchi  AUTOIMMUNITY  36-  (4)  247  -254  2003/07  [Not refereed][Not invited]
   

  Subpopulations of infiltrating lymphocytes, professional antigen-presenting cells (APCs), and Th1/Th2 cytokines that could initiate an autoimmune sialodacryoadenitis were studied in the IQI/Jic mouse model of primary Sjogren's syndrome. Although lymphocytic infiltrations were first seen in submandibular glands (SMGs) of females and in lacrimal glands (LGs) of males at 8 weeks of age, clusters of MHC class II+ , CD11c(+) , B7-2(+) dendritic cells (DCs) were already localized in these tissues at 4 weeks. At 8 weeks, the infiltrating lymphocytes consisted of almost equal numbers of B cells and CD4(+) T cells. In the inflammatory foci, MHC class II+ , CD11c(+) , B7-2(+) DCs formed network-like structures. Duct cells in the lesions showed immunoreactivities for MHC class II and ALCAM (a costimulatory adhesion molecule). IL-12 and IFN-gamma transcripts were detected by RT-PCR in SMGs of females and in LGs of males at 8-12 weeks. These results suggest that the clustered DCs might play an important role in the initiation of the adenitis, and further suggest that the DCs and epithelial cells may participate in the activation of CD4(+) T cells. It is also likely that Th1 cytokines mediate the functional interactions between the APCs and CD4(+) T cells in the early lesions.
• 超音波検査を過小評価してはいないか?

  TAKIGUCHI MITSUYOSHI  日本獣医循環器学会抄録  78th-  126  -126  2003/06/21  [Not refereed][Not invited]
  
• 最新の画像診断装置の獣医臨床応用の可能性‐超音波検査を過小評価してはいないか?

  TAKIGUCHI MITSUYOSHI  獣医麻酔外科学雑誌  34-  (Supplement 1)  140  -140  2003/06/10  [Not refereed][Not invited]
  
• Moderator band cardiomyopathyを疑った猫の一例

  濱本 耕平, 滝口 満喜, 古橋 由美子, 高橋 徹  日本獣医循環器学会抄録  78回-  210  -211  2003/06
• Cloning and polymorphic analysis of canine 21-hydroxyrase gene: a candidate gene for alopecia X in dogs

  TAKADA KENSUKE, KITAMURA HIROSHI, TAKIGUCHI MITSUYOSHI, SAITO MASAYUKI, HASHIMOTO AKIRA, INABA MUTSUMI  獣医生化学  40-  (1)  21  -27  2003/04/30  [Not refereed][Not invited]
  
• イヌ脱毛症Xの原因候補遺伝子：21-ヒドロキシラーゼ遺伝子のクローニングと多型解析（共著）

  獣医生化学  40(1): 21-27-  2003  [Not refereed][Not invited]
  
• 犬の消化管内異物の超音波画像診断

  Small Animal Clinic  133: 22-25-  2003  [Not refereed][Not invited]
  
• Cloning of canine 21-hydroxylase gene and its polymorphic analysis as a candidate gene for congenital adrenal hyperplasia-like syndrome in Pomeranians

  K Takada, H Kitamura, M Takiguchi, M Saito, A Hashimoto  RESEARCH IN VETERINARY SCIENCE  73-  (2)  159  -163  2002/10  [Not refereed][Not invited]
   

  A Strong breed predisposition to canine congenital adrenal hyperplasia-like syndrome (CAH-LS) exists and implies there is an hereditary cause. Pomeranians, one of the most predisposed breeds, have been reported to have partial adrenal dysfunction that could be associated with the pathogenesis of CAH-LS. In this study, the full-length complementary DNA (CDNA) of canine 21-OH, whose mutations are most likely responsible for the adrenal dysfunction in Pomeranians, was initially obtained from adrenal gland RNA by reverse transcription-polymerase chain reaction (RT-PCR), and its nucleotide sequence was determined. Genomic DNA samples from 16 Pomeranians and 30 control dogs, of seven different breeds, were tested for 21-OH gene polymorphisms by direct sequencing. No mutations affecting the primary structure and the transcriptional level of the 21-OH enzyme were found in Pomeranians. Our results indicate that the adrenal dysfunction in Pomeranians is not due to mutations of the 21-OH gene. This contrasts with the human form of CAH where 21-OH gene mutations are the major cause of the disease. (C) 2002 Elsevier Science Ltd. All rights reserved.
• Cloning of canine 21-hydroxylase gene and its polymorphic analysis as a candidate gene for congenital adrenal hyperplasia-like syndrome in Pomeranians

  K Takada, H Kitamura, M Takiguchi, M Saito, A Hashimoto  RESEARCH IN VETERINARY SCIENCE  73-  (2)  159  -163  2002/10  [Not refereed][Not invited]
   

  A Strong breed predisposition to canine congenital adrenal hyperplasia-like syndrome (CAH-LS) exists and implies there is an hereditary cause. Pomeranians, one of the most predisposed breeds, have been reported to have partial adrenal dysfunction that could be associated with the pathogenesis of CAH-LS. In this study, the full-length complementary DNA (CDNA) of canine 21-OH, whose mutations are most likely responsible for the adrenal dysfunction in Pomeranians, was initially obtained from adrenal gland RNA by reverse transcription-polymerase chain reaction (RT-PCR), and its nucleotide sequence was determined. Genomic DNA samples from 16 Pomeranians and 30 control dogs, of seven different breeds, were tested for 21-OH gene polymorphisms by direct sequencing. No mutations affecting the primary structure and the transcriptional level of the 21-OH enzyme were found in Pomeranians. Our results indicate that the adrenal dysfunction in Pomeranians is not due to mutations of the 21-OH gene. This contrasts with the human form of CAH where 21-OH gene mutations are the major cause of the disease. (C) 2002 Elsevier Science Ltd. All rights reserved.
• 急性特発性多発性神経根炎が疑われた犬の一例

  ITO HIROSHI, HAMAMOTO KOHEI, TAKIGUCHI MITSUYOSHI, OCHIAI KENJI  北海道獣医師会雑誌  46-  (8)  292  -292  2002/08/01  [Not refereed][Not invited]
  
• The Immediate Early Gene of Canine Herpesvirus is Transcribed through Early and Late Phases

  MIYOSHI Masahiro, OKAZAKI Katsunori, TAKIGUCHI Mitsuyoshi, KIDA Hiroshi, HASHIMOTO Akira  Journal of Veterinary Medical Science  64-  (7)  627  -631  2002/07/25  [Not refereed][Not invited]
   

  The immediate early (IE) gene of canine herpesvirus (CHV), homologue of the infected cell protein 4 (ICP4) gene of herpes simplex virus 1, is transcribed as a 4.9kb mRNA during IE phase. The IE gene was further transcribed as a 4.8kb mRNA through early (E) and late (L) phases of productive infection. Transcription of the 4.8kb mRNA initiated from downstream of the TATA box in an intron which was spliced out during IE phase. The reverse transcription-polymerase chain reaction revealed that the IE promoter was turned off during L phase at a permissive temperature. We, thus, propose to redesignate the IE gene of CHV as CICP4 gene.
• The immediate early gene of canine herpesvirus is transcribed through early and late phases

  M Miyoshi, K Okazaki, M Takiguchi, H Kida, A Hashimoto  JOURNAL OF VETERINARY MEDICAL SCIENCE  64-  (7)  627  -631  2002/07  [Not refereed][Not invited]
   

  The immediate early (IE) gene of canine herpesvirus (CHV), homologue of the infected cell protein 4 (ICP4) gene of herpes simplex virus 1, is transcribed as a 4.9kb mRNA during IE phase. The IE gene was further transcribed as a 4.8kb mRNA through early (E) and late (L) phases of productive infection. Transcription of the 4.8kb mRNA initiated from downstream of the TATA box in an intron which was spliced out during IE phase. The reverse transcription-polymerase chain reaction revealed that the IE promoter was turned off during L phase at a permissive temperature. We, thus, propose to redesignate the IE gene of CHV as CICP4 gene.
• Rhabdomyosarcoma (botryoid sarcoma) of the urinary bladder in a Maltese

  M Takiguchi, T Watanabe, H Okada, T Kudo, K Yamada, J Yasuda, A Hashimoto  JOURNAL OF SMALL ANIMAL PRACTICE  43-  (6)  269  -271  2002/06  [Not refereed][Not invited]
   

  A urinary bladder tumour was diagnosed in a two-year-old female Maltese with haematuria and pollakluria on the basis of ultrasonography and pneumocystography findings. The mass was resected, and the bladder was preserved at surgery. Histological and immunohistochemical examination confirmed the tumour to be a rhabdomyosarcoma, which has rarely been reported in small breeds of dog. There was no recurrence of the tumour at the original site in the urinary bladder two months later, when the dog died due to metastasis to the liver. This is believed to be the first report of bladder rhabdomyosarcoma in a Maltese.
• Rhabdomyosarcoma (botryoid sarcoma) of the urinary bladder in a Maltese

  M Takiguchi, T Watanabe, H Okada, T Kudo, K Yamada, J Yasuda, A Hashimoto  JOURNAL OF SMALL ANIMAL PRACTICE  43-  (6)  269  -271  2002/06  [Not refereed][Not invited]
   

  A urinary bladder tumour was diagnosed in a two-year-old female Maltese with haematuria and pollakluria on the basis of ultrasonography and pneumocystography findings. The mass was resected, and the bladder was preserved at surgery. Histological and immunohistochemical examination confirmed the tumour to be a rhabdomyosarcoma, which has rarely been reported in small breeds of dog. There was no recurrence of the tumour at the original site in the urinary bladder two months later, when the dog died due to metastasis to the liver. This is believed to be the first report of bladder rhabdomyosarcoma in a Maltese.
• 犬微小ウイルス(犬パルボウイルス1型)分離株の性状と血清疫学

  HASHIMA TAKAYUKI, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA, UNE YUMI, SASAKI NOBUO, TOOYA YUKINOBU, ONISHI TAKAFUMI, MAEDA TAKESHI, CARMICHAEL L E  日本獣医学会学術集会講演要旨集  133rd-  106  -106  2002/03/01  [Not refereed][Not invited]
  
• 肝皮膚症候群と診断された犬の肝臓超音波画像

  Small Animal Clinic  129: 29-31-  2002  [Not refereed][Not invited]
  
• Association of canine minute virus(canine parvovirus type Ⅰ)with diseased puppies in Japan.

  J. Clin. Microbiol.  40-  3993  -3998  2002  [Not refereed][Not invited]
  
• Association of canine minute virus(canine parvovirus type Ⅰ)with diseased puppies in Japan. "jointly worked"

  J. Clin. Microbiol.  40(11);3993-3998-  2002  [Not refereed][Not invited]
  
• 犬のすい臓疾患におけるトリプシン様免疫活性(Trypsin‐like immunoreactivity;TLI)測定の診断的意義 II 急性すい炎の病態とTLI

  TAKIGUCHI MITSUYOSHI  動物臨床医学会年次大会プロシーディング  22nd-  (3)  181  -183  2001/11/16  [Not refereed][Not invited]
  
• A serological survey of minute virus of canines (MVC; Canine parvovirus type-1) in dogs in the Tokai area of Japan

  A Hashimoto, M Takiguchi, K Hirai, H Kida, LE Carmichael  JAPANESE JOURNAL OF VETERINARY RESEARCH  49-  (3)  249  -253  2001/11  [Not refereed][Not invited]
   

  A serological survey for antibodies to minute virus of canines (WC) by use of a hemagglutination-inhibition (HI) test was performed on sera collected from dogs in the Tokai area of Japan. Forty-one of 266 (15.4%) sera had positive titers of 1 : 40 or higher against the MVC. Results suggest that WC may have been present in dogs in Japan since, at least, 1990. From this serosurvey, WC appears to be established in the dog population in Japan. MVC may have a role as a newly recognized viral pathogen of dogs in Japan.
• A serological survey of minute virus of canines (MVC; Canine parvovirus type-1) in dogs in the Tokai area of Japan

  A Hashimoto, M Takiguchi, K Hirai, H Kida, LE Carmichael  JAPANESE JOURNAL OF VETERINARY RESEARCH  49-  (3)  249  -253  2001/11  [Not refereed][Not invited]
   

  A serological survey for antibodies to minute virus of canines (WC) by use of a hemagglutination-inhibition (HI) test was performed on sera collected from dogs in the Tokai area of Japan. Forty-one of 266 (15.4%) sera had positive titers of 1 : 40 or higher against the MVC. Results suggest that WC may have been present in dogs in Japan since, at least, 1990. From this serosurvey, WC appears to be established in the dog population in Japan. MVC may have a role as a newly recognized viral pathogen of dogs in Japan.
• Bovine serum adenosine deaminase activity and inflammatory change of the liver.

  YASUDA JUN, TAKUMA SHUNSUKE, TAKIGUCHI MITSUYOSHI, OKADA KEIJI, HASHIMOTO AKIRA  獣医生化学  38-  (2)  33  -37  2001/10/31  [Not refereed][Not invited]
  
• ウマヘルペスウイルス9型(EHV‐9)経鼻接種によるシリアンハムスター脳炎モデルのMRIによる経時的観察

  SUZUKI KENGO, ASANUMA TAKETOSHI, FUKUSHI HIDETO, HIRAI KATSUYA, TAKIGUCHI MITSUYOSHI, KUWAHARA MIKINORI, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  132nd-  221  -221  2001/09/07  [Not refereed][Not invited]
  
• 非定形型副腎皮質機能低下症の犬の一例

  ITO KEI, TAKIGUCHI MITSUYOSHI, TAKADA KENSUKE, HAMAMOTO KOHEI, HASHIMOTO AKIRA  北海道獣医師会雑誌  45-  (8)  272  -272  2001/08/01  [Not refereed][Not invited]
  
• 副腎皮質機能低下の猫の一例

  WATANABE MINAKO, TAKIGUCHI MITSUYOSHI, TAKADA KENSUKE, HAMAMOTO KOHEI, HASHIMOTO AKIRA  北海道獣医師会雑誌  45-  (8)  272  -272  2001/08/01  [Not refereed][Not invited]
  
• Malignant seminoma with systemic metastases in a dog

  M Takiguchi, T Iida, T Kudo, A Hashimoto  JOURNAL OF SMALL ANIMAL PRACTICE  42-  (7)  360  -362  2001/07  [Not refereed][Not invited]
   

  A three-year-old male crossbred dog was presented with multiple cutaneous nodules on the dorsum. Unilateral testicular enlargement ensued. Histopathological examination of testicular tissue and the skin lesions revealed a malignant seminoma. Despite the initiation of adjuvant chemotherapy, the dog died three months later due to systemic metastases of the seminoma to the skin, scrotum, eyes, liver, kidney and peritoneum.
• Malignant seminoma with systemic metastases in a dog

  M Takiguchi, T Iida, T Kudo, A Hashimoto  JOURNAL OF SMALL ANIMAL PRACTICE  42-  (7)  360  -362  2001/07  [Not refereed][Not invited]
   

  A three-year-old male crossbred dog was presented with multiple cutaneous nodules on the dorsum. Unilateral testicular enlargement ensued. Histopathological examination of testicular tissue and the skin lesions revealed a malignant seminoma. Despite the initiation of adjuvant chemotherapy, the dog died three months later due to systemic metastases of the seminoma to the skin, scrotum, eyes, liver, kidney and peritoneum.
• ぼうこう・尿道疾患のX線・超音波診断

  TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA  獣医麻酔外科学雑誌  32-  (Supplement 1)  67  -68  2001/07/01  [Not refereed][Not invited]
  
• 犬の副腎褐色細胞腫の超音波診断

  TAKIGUCHI MITSUYOSHI, HAMAMOTO KOHEI, TAKADA KENSUKE, HASHIMOTO AKIRA  獣医麻酔外科学雑誌  32-  (Supplement 1)  291  -292  2001/07/01  [Not refereed][Not invited]
  
• 自己免疫疾患の遺伝子治療

  TAKIGUCHI MITSUYOSHI  日本獣医学会学術集会講演要旨集  131st-  37  -37  2001/03/05  [Not refereed][Not invited]
  
• すい管結さつマウスモデルを用いたすい臓におけるc‐metの発現動態

  HAMAMOTO KOHEI, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  131st-  142  -142  2001/03/05  [Not refereed][Not invited]
  
• 犬微小ウイルス(犬パルボウイルス1型)の遺伝子の検出とウイルス分離

  MOCHIZUKI MASAMI, HASHIMOTO MICHIRU, HASHIMOTO AKIRA, TAKIGUCHI MITSUYOSHI, CARMICHAEL L E, PARRISH C R  日本獣医学会学術集会講演要旨集  131st-  81  -81  2001/03/05  [Not refereed][Not invited]
  
• 膵管結紮マウスモデルを用いた膵臓におけるc-metの発現動態

  濱本 耕平, 滝口 満喜, 橋本 晃  日本獣医学会学術集会講演要旨集  131回-  142  -142  2001/03  [Not refereed][Not invited]
  
• An improved intravesical model using human bladder cancer cell lines to optimize gene and other therapies

  T Watanabe, N Shinohara, A Sazawa, T Harabayashi, Y Ogiso, T Koyanagi, M Takiguchi, A Hashimoto, N Kuzumaki, M Yamashita, M Tanaka, HB Grossman, WF Benedict  CANCER GENE THERAPY  7-  (12)  1575  -1580  2000/12  [Not refereed][Not invited]
   

  Orthotopic implantation of human bladder cancer cells into immunodeficient mice is an important tool for studying the biology and effects of therapy. Nevertheless, the incidence of tumor implantation and growth by transurethral instillation of the human bladder cancer cells into murine bladders has been low or not reproducible. However, using a modified intravesical technique and the human bladder cancer cell lines, KU-7 and UM-UC-2, we have been able to obtain a high and reproducible incidence of superficial bladder tumors. Furthermore, intravesical administration of the LacZ adenovirus vector resulted in significant beta -galactosidase expression in these bladder tumors as well as the normal urothelium, which was associated with the removal of the glycosoaminoglycan layer. Because this modified technique produces a high incidence of superficial human tumor growth acid allows the efficacy of ge ne transfer to be evaluated, it should be a useful model for the study of intravesical gene therapy for human bladder cancer.
• An improved intravesical model using human bladder cancer cell lines to optimize gene and other therapies

  T Watanabe, N Shinohara, A Sazawa, T Harabayashi, Y Ogiso, T Koyanagi, M Takiguchi, A Hashimoto, N Kuzumaki, M Yamashita, M Tanaka, HB Grossman, WF Benedict  CANCER GENE THERAPY  7-  (12)  1575  -1580  2000/12  [Not refereed][Not invited]
   

  Orthotopic implantation of human bladder cancer cells into immunodeficient mice is an important tool for studying the biology and effects of therapy. Nevertheless, the incidence of tumor implantation and growth by transurethral instillation of the human bladder cancer cells into murine bladders has been low or not reproducible. However, using a modified intravesical technique and the human bladder cancer cell lines, KU-7 and UM-UC-2, we have been able to obtain a high and reproducible incidence of superficial bladder tumors. Furthermore, intravesical administration of the LacZ adenovirus vector resulted in significant beta -galactosidase expression in these bladder tumors as well as the normal urothelium, which was associated with the removal of the glycosoaminoglycan layer. Because this modified technique produces a high incidence of superficial human tumor growth acid allows the efficacy of ge ne transfer to be evaluated, it should be a useful model for the study of intravesical gene therapy for human bladder cancer.
• Canine extracutaneous mast-cell tumours consisting of connective tissue mast cells

  N Iwata, K Ochiai, T Kadosawa, M Takiguchi, T Umemura  JOURNAL OF COMPARATIVE PATHOLOGY  123-  (4)  306  -310  2000/11  [Not refereed][Not invited]
   

  (T)wo cases of canine extracutaneous mast-cell tumours were encountered, originating from the mucosa of either the oral cavity or the small intestine. The dogs had no neoplasms in the skin. Immunohistochemical and ultrastructural studies demonstrated that the neoplastic cells had the features of connective tissue mast cells. It would seem, therefore, that at least some extracutaneous forms of the neoplasm originate from connective tissue mast cells. Heparin was a useful cytological marker to diagnose this type of mast-cell tumour. (C) 2000 Harcourt Publishers Ltd.
• Canine extracutaneous mast-cell tumours consisting of connective tissue mast cells

  N Iwata, K Ochiai, T Kadosawa, M Takiguchi, T Umemura  JOURNAL OF COMPARATIVE PATHOLOGY  123-  (4)  306  -310  2000/11  [Not refereed][Not invited]
   

  (T)wo cases of canine extracutaneous mast-cell tumours were encountered, originating from the mucosa of either the oral cavity or the small intestine. The dogs had no neoplasms in the skin. Immunohistochemical and ultrastructural studies demonstrated that the neoplastic cells had the features of connective tissue mast cells. It would seem, therefore, that at least some extracutaneous forms of the neoplasm originate from connective tissue mast cells. Heparin was a useful cytological marker to diagnose this type of mast-cell tumour. (C) 2000 Harcourt Publishers Ltd.
• イヌの成長ホルモン反応性脱毛症における21‐ヒドロキシラーゼ遺伝子の検討

  TAKADA KENSUKE, KITAMURA HIROSHI, TAKIGUCHI MITSUYOSHI, SAITO MASAYUKI, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  130th-  123  2000/09/07  [Not refereed][Not invited]
  
• 新しいぼうこう癌同所性移植モデルを用いた経尿道的アデノウイルスベクター注入による遺伝子治療の有用性の検討

  WATANABE TAKAFUMI, SHINOHARA NOBUO, SAZAWA YO, OGISO YOSHIFUMI, TAKIGUCHI MITSUYOSHI, KUZUMAKI SUSUMU, YAMASHITA MOTOYUKI, TANAKA MOTOMIKI, GROSSMAN H B  日本獣医学会学術集会講演要旨集  130th-  27  2000/09/07  [Not refereed][Not invited]
  
• 蛋白喪失性腸症に対する酢酸オクトレオチドによる新しい治療法の試み

  TAKIGUCHI MITSUYOSHI, HAMAMOTO KOHEI, HASHIMOTO AKIRA  北海道獣医師会雑誌  44-  (7)  257  -257  2000/07/01  [Not refereed][Not invited]
  
• 2‐クロロデオキシアデノシン(2‐CdA)を用いたネコ縦隔型リンパ腫に対する新しい治療法の可能性

  FUJIKI MEGUMI, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA  北海道獣医師会雑誌  44-  (7)  244  -244  2000/07/01  [Not refereed][Not invited]
  
• CTLA4IgG gene delivery prevents autoantibody production and lupus nephritis in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, A Hashimoto, T Uede  FASEB JOURNAL  14-  (6)  A1207  -A1207  2000/04  [Not refereed][Not invited]
  
• Significance of the Grb2 and Son of sevenless (Sos) proteins in human bladder cancer cell lines

  T Watanabe, N Shinohara, K Moriya, A Sazawa, Y Kobayashi, Y Ogiso, M Takiguchi, J Yasuda, T Koyanagi, N Kuzumaki, A Hashimoto  IUBMB LIFE  49-  (4)  317  -320  2000/04  [Not refereed][Not invited]
   

  The epidermal growth factor (EGF) receptor has been suggested to have an important role in tumor initiation and progression of human bladder cancers. Grb2 protein, which is the downstream effector of the EGF receptor, acts as an adaptor protein between the EGF receptor and the Ras guanine-nucleotide exchange factor, son of sevenless (Sos) protein. Sos protein regulates the action of Ras protein by promoting the exchange of GDP for GTP, However, the significance of Grb2 and Sos proteins, which is related to EGF-triggered Ras activation, has not, been elucidated in human bladder cancer. The aim of the present study is to clarify the significance of these proteins in human bladder cancer cell lines. In the present study, we used four human bladder cancer cell lines (T24, KU-7, UMUC-2, UMUC-6) and two kinds of cultured normal urothelial cells (HMKU-1, HMKU-2) isolated from patients with no malignancy. We examined the expression of EGF receptor, Grb2, and Sos proteins in these cells by Western blot analysis. Furthermore, the bladder cancer cell lines were subjected to sequence analysis to identify a point mutation in the c-H-ras gene at codon 12. There was no marked difference in the expression of the EGF receptor between human bladder cancer cell lines and cultured normal urothelial cells. On the other hand, expression of Grb2 and Sos proteins was substantially increased in all human bladder cancer cell lines examined in comparison with cultured normal urothelial cells, whether codon 12 of H-ras was mutated or not. These results suggest that the amplification of both Grb2 and SOS proteins plays an important role in the carcinogenesis of human bladder cancer.
• Significance of the Grb2 and Son of sevenless (Sos) proteins in human bladder cancer cell lines

  T Watanabe, N Shinohara, K Moriya, A Sazawa, Y Kobayashi, Y Ogiso, M Takiguchi, J Yasuda, T Koyanagi, N Kuzumaki, A Hashimoto  IUBMB LIFE  49-  (4)  317  -320  2000/04  [Not refereed][Not invited]
   

  The epidermal growth factor (EGF) receptor has been suggested to have an important role in tumor initiation and progression of human bladder cancers. Grb2 protein, which is the downstream effector of the EGF receptor, acts as an adaptor protein between the EGF receptor and the Ras guanine-nucleotide exchange factor, son of sevenless (Sos) protein. Sos protein regulates the action of Ras protein by promoting the exchange of GDP for GTP, However, the significance of Grb2 and Sos proteins, which is related to EGF-triggered Ras activation, has not, been elucidated in human bladder cancer. The aim of the present study is to clarify the significance of these proteins in human bladder cancer cell lines. In the present study, we used four human bladder cancer cell lines (T24, KU-7, UMUC-2, UMUC-6) and two kinds of cultured normal urothelial cells (HMKU-1, HMKU-2) isolated from patients with no malignancy. We examined the expression of EGF receptor, Grb2, and Sos proteins in these cells by Western blot analysis. Furthermore, the bladder cancer cell lines were subjected to sequence analysis to identify a point mutation in the c-H-ras gene at codon 12. There was no marked difference in the expression of the EGF receptor between human bladder cancer cell lines and cultured normal urothelial cells. On the other hand, expression of Grb2 and Sos proteins was substantially increased in all human bladder cancer cell lines examined in comparison with cultured normal urothelial cells, whether codon 12 of H-ras was mutated or not. These results suggest that the amplification of both Grb2 and SOS proteins plays an important role in the carcinogenesis of human bladder cancer.
• ぼうこう内腫りゅう性病変の画像所見を示したぼうこう血腫の1例

  WATANABE TAKAFUMI, TAKIGUCHI MITSUYOSHI, KUDO TADAAKI, HASHIMOTO AKIRA  北海道獣医師会雑誌  44-  (4)  104  -106  2000/04/01  [Not refereed][Not invited]
  
• イヌヘルペスウイルス(CHV)感染新生子犬の脳病変におけるウイルス抗原及びウイルスゲノムの分布

  NISHINO AKI, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA, OKAZAKI KATSUNORI, KIDA HIROSHI  日本獣医学会学術集会講演要旨集  129回-  47  -47  2000/03  [Not refereed][Not invited]
  
• CTLA4IgG gene delivery prevents autoantibody production and lupus nephritis in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, Saito, I, A Hashimoto, T Uede  LIFE SCIENCES  66-  (11)  991  -1001  2000/02  [Not refereed][Not invited]
   

  MRL/MP-lpr/lpr (MRL/lpr) mice spontaneously develop an autoimmune syndrome closely resembling systemic lupus erythematosus (SLE) in humans, characterized by hypergammaglobulinemia, various autoantibody production, and the development of fatal glomerulonephritis. We have previously demonstrated that systemic administration of soluble form of CTLA4IgG prevented autoantibody-related diseases in MRL/lpr mice. To test the potential protective effects of CTLA4IgG gene delivery on the development of lupus nephritis, we injected MRL/lpr mice with, a recombinant adenovirus vector containing CTLA4IgG gene, Adex1CACTLA4IgG (AdCTLA4IgG). It was demonstrated that a single administration of intravenous injection of AdCTLA4IgG into MRL/lpr mice resulted in almost complete amelioration of lupus nephritis.
• Suppressive effects of dominant negative ras mutant N116Y on transformed phenotypes of human bladder cancer cells

  T Watanabe, N Shinohara, A Sazawa, Y Kobayashi, Y Ogiso, T Kimura, M Takiguchi, J Yasuda, A Hashimoto, T Koyanagi, N Kuzumaki  CANCER LETTERS  149-  (1-2)  195  -202  2000/02  [Not refereed][Not invited]
   

  To investigate the suppressive effect of dominant negative H-ras mutant N116Y on transformed phenotypes, we established two N116Y ras mutant stable transfectant clones (C5, C13) of human bladder cancer cell line, UMUC-2. These N116Y ras mutant transfectants, especially the C5 cells, showed a dramatic change of cellular morphology and significantly reduced growth in soft agar compared to their control. Furthermore, phosphorylation of the Jun NH2-terminal kinase (JNK) was significantly decreased in these transfectants compared to the control. These results suggest that the N116Y-induced suppression of transformed phenotypes in UMUC-2 cells is associated with inhibition of JNK phosphorylation, (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.
• CTLA4IgG gene delivery prevents autoantibody production and lupus nephritis in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, Saito, I, A Hashimoto, T Uede  LIFE SCIENCES  66-  (11)  991  -1001  2000/02  [Not refereed][Not invited]
   

  MRL/MP-lpr/lpr (MRL/lpr) mice spontaneously develop an autoimmune syndrome closely resembling systemic lupus erythematosus (SLE) in humans, characterized by hypergammaglobulinemia, various autoantibody production, and the development of fatal glomerulonephritis. We have previously demonstrated that systemic administration of soluble form of CTLA4IgG prevented autoantibody-related diseases in MRL/lpr mice. To test the potential protective effects of CTLA4IgG gene delivery on the development of lupus nephritis, we injected MRL/lpr mice with, a recombinant adenovirus vector containing CTLA4IgG gene, Adex1CACTLA4IgG (AdCTLA4IgG). It was demonstrated that a single administration of intravenous injection of AdCTLA4IgG into MRL/lpr mice resulted in almost complete amelioration of lupus nephritis.
• Suppressive effects of dominant negative ras mutant N116Y on transformed phenotypes of human bladder cancer cells

  T Watanabe, N Shinohara, A Sazawa, Y Kobayashi, Y Ogiso, T Kimura, M Takiguchi, J Yasuda, A Hashimoto, T Koyanagi, N Kuzumaki  CANCER LETTERS  149-  (1-2)  195  -202  2000/02  [Not refereed][Not invited]
   

  To investigate the suppressive effect of dominant negative H-ras mutant N116Y on transformed phenotypes, we established two N116Y ras mutant stable transfectant clones (C5, C13) of human bladder cancer cell line, UMUC-2. These N116Y ras mutant transfectants, especially the C5 cells, showed a dramatic change of cellular morphology and significantly reduced growth in soft agar compared to their control. Furthermore, phosphorylation of the Jun NH2-terminal kinase (JNK) was significantly decreased in these transfectants compared to the control. These results suggest that the N116Y-induced suppression of transformed phenotypes in UMUC-2 cells is associated with inhibition of JNK phosphorylation, (C) 2000 Elsevier Science Ireland Ltd. All rights reserved.
• 膀胱内腫瘤性病変の画像所見を示した膀胱血腫の1例(共著)

  渡邉 孝文, 滝口 満喜, 工藤 忠明, 橋本 晃  北海道獣医師会雑誌  44-  (4)  104  -106  2000  [Not refereed][Not invited]
  
• Involvement of CD28/CTLA4-B7 costimulatory pathway in the development of lymphadenopathy and splenomegaly in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, MM Rashid, N Tosa, S Chikuma, A Hashimoto, T Uede  JOURNAL OF VETERINARY MEDICAL SCIENCE  62-  (1)  29  -36  2000/01  [Not refereed][Not invited]
   

  MRL/lpr mouse is an established animal model which develops autoimmune diseases including glomerulonephritis, sialoadenitis, hepatitis and inflammatory lung disease. Additionally, it has been reported that lpr strains uniquely accumulate CD3(+)CD4(-)CD8(-)B220(+) (double negative, DN) T cells in lymphoid organs leading to lymphadenopathy and splenomegaly. To investigate the role of CD28/CTLA4-B7 pathway in the development of lymphadenopathy and splenomegaly, MRL/lpr mice were treated with soluble form of CTLA4 molecules. CTLA4IgG, which efficiently blocks this pathway. It was demonstrated that (i) the development of DN T cells was independent of the CD28/CTLA4-B7 pathway, (ii) the CD28/CTLA4-B7 pathway was required for the development of lymphadenopathy and splenomegaly, (iii) the CD28/CTLA4-B7 pathway was important for the accumulation of various cell populations in the lymph node and spleen, (iv) composition of the accumulating cell populations was not altered by CTLA4IgG treatment, and (v) activation of conventional T cells and IL-4 production from conventional T cells were the CD28/CTLA4-B7 pathway dependent. Thus, we concluded that the CD28/CTLA4-B7 pathway was required for the development of full-blown lymphadenopathy and splenomegaly in MRL/lpr mice.
• Structure of the infected cell protein 0 gene of canine herpesvirus

  M Miyoshi, M Takiguchi, J Yasuda, A Hashimoto, A Takada, K Okazaki, H Kida  ARCHIVES OF VIROLOGY  145-  (8)  1715  -1723  2000  [Not refereed][Not invited]
   

  The canine herpesvirus infected cell protein 0 (CICP0) gene was sequenced. The CICP0 gene was transcribed as a 1.4 kb mRNA from the end of the unique long region nearby the internal repeat during early phase of productive infection of the virus. An open reading frame of the gene encodes a polypeptide of 333 amino acids. The RING finger domain and acidic transcriptional activation domain were found at the N-terminus and within the middle region in the deduced amino acid sequence, respectively, suggesting that the CICP0, like the ICP0 of herpes simplex virus 1, is a transactivating protein.
• Involvement of CD28/CTLA4-B7 costimulatory pathway in the development of lymphadenopathy and splenomegaly in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, MM Rashid, N Tosa, S Chikuma, A Hashimoto, T Uede  JOURNAL OF VETERINARY MEDICAL SCIENCE  62-  (1)  29  -36  2000/01  [Not refereed][Not invited]
   

  MRL/lpr mouse is an established animal model which develops autoimmune diseases including glomerulonephritis, sialoadenitis, hepatitis and inflammatory lung disease. Additionally, it has been reported that lpr strains uniquely accumulate CD3(+)CD4(-)CD8(-)B220(+) (double negative, DN) T cells in lymphoid organs leading to lymphadenopathy and splenomegaly. To investigate the role of CD28/CTLA4-B7 pathway in the development of lymphadenopathy and splenomegaly, MRL/lpr mice were treated with soluble form of CTLA4 molecules. CTLA4IgG, which efficiently blocks this pathway. It was demonstrated that (i) the development of DN T cells was independent of the CD28/CTLA4-B7 pathway, (ii) the CD28/CTLA4-B7 pathway was required for the development of lymphadenopathy and splenomegaly, (iii) the CD28/CTLA4-B7 pathway was important for the accumulation of various cell populations in the lymph node and spleen, (iv) composition of the accumulating cell populations was not altered by CTLA4IgG treatment, and (v) activation of conventional T cells and IL-4 production from conventional T cells were the CD28/CTLA4-B7 pathway dependent. Thus, we concluded that the CD28/CTLA4-B7 pathway was required for the development of full-blown lymphadenopathy and splenomegaly in MRL/lpr mice.
• Structure of the infected cell protein O gene of canine herpesvirus. "jointly worked"

  Arch. Virol.  145-  (8)  1715  -1723  2000  [Not refereed][Not invited]
  
• Development of an Ultrasonic Washing Machine for Animals and its Clinical Application to Chronic Skin Diseases of Dogs.

  YASUDA JUN, NAKAO KIICHI, SHIMIZU KYOKO, KAMATA SATORU, KITAMURA YUKITOSHI, TAKIGUCHI MITSUYOSHI  獣医畜産新報  52-  (939)  801  -804  1999/10/01  [Not refereed][Not invited]
  
• Dominant negative ras mutant N116Y発現アデノウイルスベクターを用いたぼうこう癌に対する遺伝子治療の基礎的研究

  WATANABE TAKAFUMI, SHINOHARA NOBUO, SAZAWA AKIRA, TAKIGUCHI MITSUYOSHI, KOYANAGI TOMOHIKO, KATSUMAKI SUSUMU, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  128th-  186  -186  1999/09/01  [Not refereed][Not invited]
  
• 脳包虫症ラットモデルのMRI画像観察

  MATSUMOTO YUKARI, ASANUMA TAKETOSHI, NAKAO MINORU, NAKATANI KAZUHIRO, TAKIGUCHI MITSUYOSHI, ITO RYO, KUWABARA MIKINORI, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  128th-  167  -167  1999/09/01  [Not refereed][Not invited]
  
• CTLA4IgG発現アデノウイルスベクターを用いたループス腎炎への遺伝子治療の試み

  TAKIGUCHI MITSUYOSHI, MURAKAMI MASAAKI, NAKAGAWA IZUMI, SAITO IZUMI, HASHIMOTO AKIRA, KAMIDE TOSHIMITSU  日本獣医学会学術集会講演要旨集  128th-  20  -20  1999/09/01  [Not refereed][Not invited]
  
• 犬副腎髄質褐色細胞腫の画像診断と尿中カテコールアミン代謝産物の診断的意義

  滝口 満喜, 宮田 佳代子, 橋本 晃  北海道獣医師会雑誌  43-  (8)  256  -256  1999/08
• インスリン療法により重度の溶血性貧血を呈した糖尿病性ケトアシドーシスの猫の1例

  濱本 耕平, 滝口 満喜, 橋本 晃  北海道獣医師会雑誌  43-  (8)  253  -253  1999/08
• Combination of CTLA41GG and FTY720 suppresses development of obliterative bronchiolitis and respiratory epithelial injury in a murine heterotopic airway model

  K Konishi, M Murakami, A Yamada, M Takiguchi, M Morikawa, T Omura, S Todo, T Uede  TRANSPLANTATION  67-  (9)  S605  -S605  1999/05  [Not refereed][Not invited]
  
• Detection of canine herpesvirus DNA in the ganglionic neurons and the lymph node lymphocytes of latently infected dogs

  M Miyoshi, Y Ishii, M Takiguchi, A Takada, J Yasuda, A Hashimoto, K Okazaki, H Kida  JOURNAL OF VETERINARY MEDICAL SCIENCE  61-  (4)  375  -379  1999/04  [Not refereed][Not invited]
   

  TO determine the site of latent infection of canine herpesvirus (CHV), tissues from dogs convalescent from acute infection with CHV were examined for the presence of viral genome DNA by the nested polymerase chain reaction. CHV DNA was detected in the trigeminal ganglia and the retropharyngeal lymph nodes. In situ hybridization study of the tissues revealed that CHV genome persisted in the nuclei of ganglionic neurons and lymphocytes.
• Blockade of CD28/CTLA4-B7 pathway prevented autoantibody-related diseases but not lung disease in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, A Yamada, S Chikuma, Y Kawaguchi, A Hashimoto, T Uede  LABORATORY INVESTIGATION  79-  (3)  317  -326  1999/03  [Not refereed][Not invited]
   

  We studied the role of CD28/CTLA4 costimulatory T-cell activation pathway on the pathogenesis of MRL/Ipr mice. Administration of CTLA4IgG from day 0 significantly inhibited autoantibody production such as anti-double-stranded DNA antibody and rheumatoid factor. In addition, end-organ diseases in kidney, salivary gland, and liver were significantly improved. Improvement of pathologic findings coincided with a significant improvement in survival. At 350 days of age, 90% of mice treated with CTLA4IgG from day 0 were still alive, compared with none of mice treated with hIgG. As expected, activation of conventional T cells was significantly inhibited after CTLA4IgG treatment. However, lung disease that was characterized by perivascular accumulation and interstitial infiltration of lymphocytes and macrophages was not inhibited. Even after CTLA4IgG treatment from day 0, pathologic findings of lung disease were not improved. Additionally, the expression of activation markers such as B7-1, B7-2, CD71, ICAM1, and LFA1 on Mac1(+) fraction in both spleen and lung and the concentration of TNF alpha in bronchoalveolar lavage fluid were not significantly suppressed. These results demonstrated that lung disease was independent of the CD28/CTLA4-B7 pathway. Thus, this study emphasizes the differential dependence of the CD28/CTLA4-B7 pathway in development of diseases in MRL/Ipr mice.
• Blockade of CD28/CTLA4-B7 pathway prevented autoantibody-related diseases but not lung disease in MRL/lpr mice

  M Takiguchi, M Murakami, Nakagawa, I, A Yamada, S Chikuma, Y Kawaguchi, A Hashimoto, T Uede  LABORATORY INVESTIGATION  79-  (3)  317  -326  1999/03  [Not refereed][Not invited]
   

  We studied the role of CD28/CTLA4 costimulatory T-cell activation pathway on the pathogenesis of MRL/Ipr mice. Administration of CTLA4IgG from day 0 significantly inhibited autoantibody production such as anti-double-stranded DNA antibody and rheumatoid factor. In addition, end-organ diseases in kidney, salivary gland, and liver were significantly improved. Improvement of pathologic findings coincided with a significant improvement in survival. At 350 days of age, 90% of mice treated with CTLA4IgG from day 0 were still alive, compared with none of mice treated with hIgG. As expected, activation of conventional T cells was significantly inhibited after CTLA4IgG treatment. However, lung disease that was characterized by perivascular accumulation and interstitial infiltration of lymphocytes and macrophages was not inhibited. Even after CTLA4IgG treatment from day 0, pathologic findings of lung disease were not improved. Additionally, the expression of activation markers such as B7-1, B7-2, CD71, ICAM1, and LFA1 on Mac1(+) fraction in both spleen and lung and the concentration of TNF alpha in bronchoalveolar lavage fluid were not significantly suppressed. These results demonstrated that lung disease was independent of the CD28/CTLA4-B7 pathway. Thus, this study emphasizes the differential dependence of the CD28/CTLA4-B7 pathway in development of diseases in MRL/Ipr mice.
• MRL/lprマウス肺病変形成におけるCD28/CTLA4‐B7補助シグナルの役割

  TAKIGUCHI MITSUYOSHI, MURAKAMI MASAAKI, NAKAGAWA IZUMI, YASUDA JUN, HASHIMOTO AKIRA, KAMIDE TOSHIMITSU  日本獣医学会学術集会講演要旨集  127th-  68  -68  1999/03/01  [Not refereed][Not invited]
  
• 動物用超音波洗浄装置を用いた犬の慢性皮膚疾患の治療について

  NAKAO KIICHI, YASUDA JUN, SHIMIZU KYOKO, KITAMURA YUKITOSHI, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA  日本産業動物獣医学会・日本小動物獣医学会・日本獣医公衆衛生学会年次大会  1998-  362  1999/02  [Not refereed][Not invited]
  
• X線および超音波検査で副腎髄質腫ようが疑われた犬の一例

  MIYATA KAYOKO, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA  日本産業動物獣医学会・日本小動物獣医学会・日本獣医公衆衛生学会年次大会  1998-  354  1999/02  [Not refereed][Not invited]
  
• 犬の皮膚病に対する動物用超音波洗浄装置の応用(共著)

  安田 準, 中尾 喜一, 清水 恭子, 釜田 悟, 北村 之利, 滝口 満喜  獣医畜産新報  52-  (10)  801  -804  1999  [Not refereed][Not invited]
  
• Structure of the immediate early gene of canine herpesvirus - Brief report

  M Miyoshi, M Takiguchi, J Yasuda, A Hashimoto, A Takada, K Okazaki, H Kida  ARCHIVES OF VIROLOGY  144-  (2)  407  -420  1999  [Not refereed][Not invited]
   

  The nucleotide sequence of the immediate early (IE) gene of canine herpesvirus was determined. This gene was located in the inverted repeat regions, encoding a polypeptide of 1,383 amino acids. The predicted amino acid sequence was most closely related to that of the feline herpesvirus 1 IE protein among those of other alphaherpesviruses. DNA binding and transcriptional activation domains were found in the IE protein. A spliced region of the IE gene transcript was determined in its 5' non-coding region.
• Preliminary study on clinical application of endoscopic ultrasonography (EUS) in small animal practice: EUS findings of experimentally-induced cystic lesion in the gastric wall in dogs (in Japanese with English Summary) 'jointly worked'

  Jpn. J. Vet. Imag  10-  37  -40  1999  [Not refereed][Not invited]
  
• Structure of immediate early gene of canine herpesvirus. "jointly worked"

  Arch. Virol.  144-  (2)  407  -420  1999  [Not refereed][Not invited]
  
• Detection of canine herpesvirns DNA in the ganglionic neurons and the lymph node lymphocytes of latently infected dogs. "jointly worked"

  Masahiro Miyoshi, Yuki Ishii, Mitsuyoshi Takiguchi, Ayato Takada, Jun Yasuda, Akira Hashimoto, Katsunori Okazaki, Hiroshi Kida  J. Ved. Med. Sci.  61-  (4)  375  -379  1999  [Not refereed][Not invited]
  
• Preliminary study on clinical application of endoscopic ultrasonography (EUS) in small animal practice: EUS findings of experimentally-induced cystic lesion in the gastric wall in dogs (in Japanese with English Summary) 'jointly worked'

  Jpn. J. Vet. Imag  10-  37  -40  1999  [Not refereed][Not invited]
  
• Laboratory of Pathobiology

  HASHIMOTO Akira, TAKIGUCHI Mitsuyoshi  Japanese journal of veterinary research  46-  (2)  90  -91  1998/11/30
• Endoscopic ultrasonography of the pancreas in the dog

  Y Morita, M Takiguchi, J Yasuda, T Kitamura, M Syakalima, KD Eom, A Hashimoto  VETERINARY RADIOLOGY & ULTRASOUND  39-  (6)  552  -556  1998/11  [Not refereed][Not invited]
   

  Endoscopic ultrasonography was done in 12 normal adult dogs to investigate its efficacy in visualization of the pancreas. The endoscopic ultrasonographic device used in the present study had a curved-array ultrasound transducer mounted in front of the objective lens. The tip of the ultrasonic endoscope was inserted into the stomach, and all examinations of the pancreas were performed from within the stomach. Endoscopic ultrasoraography provided good images of most parts of the pancreas except for the ends of each lobe, Useful information about the pancreatic parenchyma, including pancreatic lobular structure, pancreatic duct, and vessels of the pancreas was obtained by endoscopic ultrasonography, Blood flow within vessels was detected using color Doppler and pulsed-wave Doppler examination.. These results suggest that endoscopic ultrasonography is available as an effective diagnostic modality in small animal practice.
• Endoscopic ultrasonography of the pancreas in the dog

  Y Morita, M Takiguchi, J Yasuda, T Kitamura, M Syakalima, KD Eom, A Hashimoto  VETERINARY RADIOLOGY & ULTRASOUND  39-  (6)  552  -556  1998/11  [Not refereed][Not invited]
   

  Endoscopic ultrasonography was done in 12 normal adult dogs to investigate its efficacy in visualization of the pancreas. The endoscopic ultrasonographic device used in the present study had a curved-array ultrasound transducer mounted in front of the objective lens. The tip of the ultrasonic endoscope was inserted into the stomach, and all examinations of the pancreas were performed from within the stomach. Endoscopic ultrasoraography provided good images of most parts of the pancreas except for the ends of each lobe, Useful information about the pancreatic parenchyma, including pancreatic lobular structure, pancreatic duct, and vessels of the pancreas was obtained by endoscopic ultrasonography, Blood flow within vessels was detected using color Doppler and pulsed-wave Doppler examination.. These results suggest that endoscopic ultrasonography is available as an effective diagnostic modality in small animal practice.
• Endoscopic ultrasonography of the pancreas in the dog

  Y Morita, M Takiguchi, J Yasuda, T Kitamura, M Syakalima, KD Eom, A Hashimoto  VETERINARY RADIOLOGY & ULTRASOUND  39-  (6)  552  -556  1998/11  [Not refereed][Not invited]
   

  Endoscopic ultrasonography was done in 12 normal adult dogs to investigate its efficacy in visualization of the pancreas. The endoscopic ultrasonographic device used in the present study had a curved-array ultrasound transducer mounted in front of the objective lens. The tip of the ultrasonic endoscope was inserted into the stomach, and all examinations of the pancreas were performed from within the stomach. Endoscopic ultrasoraography provided good images of most parts of the pancreas except for the ends of each lobe, Useful information about the pancreatic parenchyma, including pancreatic lobular structure, pancreatic duct, and vessels of the pancreas was obtained by endoscopic ultrasonography, Blood flow within vessels was detected using color Doppler and pulsed-wave Doppler examination.. These results suggest that endoscopic ultrasonography is available as an effective diagnostic modality in small animal practice.
• Endoscopic ultrasonographic findings of the pancreas after pancreatic duct ligation in the dog

  Y Morita, M Takiguchi, J Yasuda, KD Eom, A Hashimoto  VETERINARY RADIOLOGY & ULTRASOUND  39-  (6)  557  -562  1998/11  [Not refereed][Not invited]
   

  Endoscopic ultrasonographic evaluation and gray-scale histogram analysis of pancreatic atrophy after pancreatic duct ligation were performed in four normal adult dogs. Using endoscopic ultrasonography, markedly dilated pancreatic ducts were visualized, and the pancreas became gradually atrophied with a hyperechoic parenchyma, In gray-scale histogram analysis of the pancreas, mean brightness increased gradually until 8 weeks, then decreased temporally. Standard deviation of the histogram increased markedly and then fluctuated until the 4th week, after which the mean brightness and standard deviation became stable. At 4 weeks postoperatively, collapse of most pancreatic acinar structures were observed, and each atrophic lobule was associated with a significantly large amount of interstitial fibrous tissue at histopathology, At 12 weeks postoperatively, most exocrine tissue had decreased and was partly replaced by fibrous and fatty tissue, These changes of mean brightness and standard deviation reflected the histologic analysis. These findings indicated that endoscopic ultrasonography is a useful technique to image such atrophic disorders of the pancreas as chronic pancreatitis. Furthermore, gray-scale histogram analysis provides helpful information for ultrasonographic tissue characterization of the pancreas.
• 1prマウス肺病変形成におけるCD28/CTLA4-B7補助シグナルの役割

  滝口 満喜, 村上 正晃, 中川 泉, 橋本 晃, 上出 利光  日本臨床免疫学会会誌  (26回抄録集)  376  -376  1998/10
• lprマウス肺病変形成におけるCD28/CTLA4‐B7補助シグナルの役割

  TAKIGUCHI MITSUYOSHI, MURAKAMI MASAAKI, NAKAGAWA IZUMI, HASHIMOTO AKIRA, UEDE TOSHIMITSU  日本免疫学会総会・学術集会記録  28-  376  1998/10  [Not refereed][Not invited]
  
• Isoenzyme Diagnosis in Veterinary Small Animal Medicine.

  YASUDA JUN, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA, SYAKALIMA M S  動物臨床医学  7-  (2)  57  -65  1998/09  [Not refereed][Not invited]
  
• 犬の血中アデノシンデアミナーゼ活性の臨床的意義について

  安田 準, 高橋 恵理子, 滝口 満喜, 橋本 晃  北海道獣医師会雑誌  42-  (8)  290  -290  1998/08
• Pathohistological finding supplement of pancreas acinus atrophy of dog.

  AZUMA YOKO, TAKIGUCHI MITSUYOSHI, YASUDA JUN, OCHIAI KENJI, KUDO TADAAKI, IWANAGA TOSHIHIKO, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  126th-  263  -263  1998/08  [Not refereed][Not invited]
  
• Concern of CD28/CTLA4-B7 support signal in pathogeny of MRL/lpr mouse.

  TAKIGUCHI MITSUYOSHI, MURAKAMI MASAAKI, NAKAGAWA IZUMI, YASUDA JUN, HASHIMOTO AKIRA, KAMIIDE TOSHIMITSU  日本獣医学会学術集会講演要旨集  126th-  22  -22  1998/08  [Not refereed][Not invited]
  
• Endoscopic and transcutaneous ultrasonographic findings and grey-scale histogram analysis in dogs with caerulein-induced pancreatitis

  Y Morita, M Takiguchi, J Yasuda, T Kitamura, M Syakalima, A Hashimoto  VETERINARY QUARTERLY  20-  (3)  89  -92  1998/07  [Not refereed][Not invited]
   

  Endoscopic ultrasonography (EUS), grey-scale histogram analysis of EUS images, and transcutaneous ultrasonography (TUS) were done in four dogs with caerulein-induced pancreatitis. One other dog was subjected to laparotomy and biopsy specimens were collected for histopathology, By EUS, the pancreatic lesions were first detected at 60 minutes after the start of caerulein infusion. They were detected after 120 to 150 minutes when using TUS, EUS findings included swelling, a more distinct lobular pattern, subcapsular hypoechoic areas, and anechoic stripes through the pancreatic tissue. No marked changes in the histogram analysis was seen until 30 minutes. From 30 to 60 minutes, a decrease in the mean brightness of the pancreatic tissue was observed. These changes in mean brightness reflected histopathological findings showing vacuolization of acinar cells and interstitial oedema of the pancreas. These findings indicated that EUS can detect slight and diffuse changes in pancreatic tissue. Furthermore, grey-scale histogram analysis detects histopathological changes more sensitively than endoscopic ultrasound images.
• Endoscopic and transcutaneous ultrasonographic findings and grey-scale histogram analysis in dogs with caerulein-induced pancreatitis

  Y Morita, M Takiguchi, J Yasuda, T Kitamura, M Syakalima, A Hashimoto  VETERINARY QUARTERLY  20-  (3)  89  -92  1998/07  [Not refereed][Not invited]
   

  Endoscopic ultrasonography (EUS), grey-scale histogram analysis of EUS images, and transcutaneous ultrasonography (TUS) were done in four dogs with caerulein-induced pancreatitis. One other dog was subjected to laparotomy and biopsy specimens were collected for histopathology, By EUS, the pancreatic lesions were first detected at 60 minutes after the start of caerulein infusion. They were detected after 120 to 150 minutes when using TUS, EUS findings included swelling, a more distinct lobular pattern, subcapsular hypoechoic areas, and anechoic stripes through the pancreatic tissue. No marked changes in the histogram analysis was seen until 30 minutes. From 30 to 60 minutes, a decrease in the mean brightness of the pancreatic tissue was observed. These changes in mean brightness reflected histopathological findings showing vacuolization of acinar cells and interstitial oedema of the pancreas. These findings indicated that EUS can detect slight and diffuse changes in pancreatic tissue. Furthermore, grey-scale histogram analysis detects histopathological changes more sensitively than endoscopic ultrasound images.
• Preliminary studies on clinical application of endoscopic ultrasonography(EUS) in small animal practice: EUS findings of experimentally-induced cystic lesion in the gastric wall in dogs.

  MORITA YUTAKA, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA  獣医画像診断  (10)  37  -40  1998/06  [Not refereed][Not invited]
  
• The canine alkaline phosphatases: A review of the isoenzymes in serum, analytical methods and their diagnostic application

  M Syakalima, M Takiguchi, J Yasuda, A Hashimoto  JAPANESE JOURNAL OF VETERINARY RESEARCH  46-  (1)  3  -11  1998/05  [Not refereed][Not invited]
   

  This paper reviews the alkaline phosphatases in canine serum, the analytical methods used for qualitative and/or quantitative detection of these isoenzymes, and the diagnostic significancy of each of these isoenzymes. The paper further describes some of the latest advances in our knowledge of the canine alkaline phosphatases and possible areas of future research.
• The canine alkaline phosphatases: A review of the isoenzymes in serum, analytical methods and their diagnostic application

  M Syakalima, M Takiguchi, J Yasuda, A Hashimoto  JAPANESE JOURNAL OF VETERINARY RESEARCH  46-  (1)  3  -11  1998/05  [Not refereed][Not invited]
   

  This paper reviews the alkaline phosphatases in canine serum, the analytical methods used for qualitative and/or quantitative detection of these isoenzymes, and the diagnostic significancy of each of these isoenzymes. The paper further describes some of the latest advances in our knowledge of the canine alkaline phosphatases and possible areas of future research.
• Blockade of CD28/CTLA4 pathway prevented almost diseases and lymphadenopathy but not lung disease in MRL/lpr mice.

  M Takiguchi, M Murakami, Nakagawa, I, A Yamada, S Chikuma, T Uede  FASEB JOURNAL  12-  (5)  A1093  -A1093  1998/03  [Not refereed][Not invited]
  
• Ultrasonographic and serologic studies of experimental cysticercosis in rats infected with Taenia taeniaeformis

  A Ito, Y Sakakibara, L Ma, K Asano, M Takiguchi, J Yasuda, A Hashimoto  PARASITE IMMUNOLOGY  20-  (3)  105  -110  1998/03  [Not refereed][Not invited]
   

  Rats experimentally infected with Taenia taeniaeformis were followed-lip until 14 weeks post inoculation with eggs (PIE) by hepatic ultrasonographic (US) image and serum antibody response analyses. Parasitic cysts could be imaged as small (2 mm in diameter) anechoic areas with or without a parenthesis-like echogenic small line from two weeks PIE. Immunoblot analysis using antigens from oncospheres (TtO), 30-day-old (TtM-30) and 300-day-old metacestodes (TtM-300) revealed that: (1) these three different developmental stages showed their own unique patterns suggesting the presence of stage-specific antigens; (2) faint IgM antibody responses to some components of TtO and TtM-30 or TtM-300 could be detected from one and two weeks PIE, respectively, and (3) IgG responses to some major components of both TtO and TtM-300, and TtM-30 were easily detected from four and five weeks PIE onwards, respectively. Both TtO and TtM (especially TtM-300) appeared to be highly useful for detection of antibody responses in experimentally infected mts. Due to the easiness in preparation of antigens, fully developed metacestodes may be the best candidate antigens for serodiagnosis. These results strongly suggest that both US image and antibody analyses using antigens from fully developed metacestodes are useful for detection of the early stage of cysticercosis in laboratory animal model.
• Ultrasonographic and serologic studies of experimental cysticercosis in rats infected with Taenia taeniaeformis

  A Ito, Y Sakakibara, L Ma, K Asano, M Takiguchi, J Yasuda, A Hashimoto  PARASITE IMMUNOLOGY  20-  (3)  105  -110  1998/03  [Not refereed][Not invited]
   

  Rats experimentally infected with Taenia taeniaeformis were followed-lip until 14 weeks post inoculation with eggs (PIE) by hepatic ultrasonographic (US) image and serum antibody response analyses. Parasitic cysts could be imaged as small (2 mm in diameter) anechoic areas with or without a parenthesis-like echogenic small line from two weeks PIE. Immunoblot analysis using antigens from oncospheres (TtO), 30-day-old (TtM-30) and 300-day-old metacestodes (TtM-300) revealed that: (1) these three different developmental stages showed their own unique patterns suggesting the presence of stage-specific antigens; (2) faint IgM antibody responses to some components of TtO and TtM-30 or TtM-300 could be detected from one and two weeks PIE, respectively, and (3) IgG responses to some major components of both TtO and TtM-300, and TtM-30 were easily detected from four and five weeks PIE onwards, respectively. Both TtO and TtM (especially TtM-300) appeared to be highly useful for detection of antibody responses in experimentally infected mts. Due to the easiness in preparation of antigens, fully developed metacestodes may be the best candidate antigens for serodiagnosis. These results strongly suggest that both US image and antibody analyses using antigens from fully developed metacestodes are useful for detection of the early stage of cysticercosis in laboratory animal model.
• Identification of the latent infected part of canine herpesvirus.

  MIYOSHI MASAHIRO, ISHII YUKI, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA, TAKADA AYAHITO, OKAZAKI KATSUNORI, KIDA HIROSHI  日本獣医学会学術集会講演要旨集  125回-  140  -140  1998/03  [Not refereed][Not invited]
  
• Trial of quantitative analysis of alkaline phosphatase isoenzyme in canine.

  SYAKALIMA M, YASUDA JUN, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  125th-  271  -271  1998/03  [Not refereed][Not invited]
  
• Application of serum trypsin-like immunoactivity (TLI) measurement to dog pancreas external hyposecretion.

  森田豊, 滝口満喜, 安田準, 橋本晃  日本産業動物獣医学会・日本小動物獣医学会・日本獣医公衆衛生学会年次大会  1997-  292  -293  1998/02  [Not refereed][Not invited]
  
• 超音波内視鏡検査(EUS)の小動物臨床への応用に関する研究－イヌの実験的胃壁嚢胞性病変のEUS画像(予報)（共著）

  獣医画像診断  10: 37-40-  1998  [Not refereed][Not invited]
  
• 小動物臨床におけるアイソザイムの診断（共著）

  動物臨床医学  7: 57-65-  1998  [Not refereed][Not invited]
  
• Comparison of attenuation and liver-kidney contrast of liver ultrasonographs with histology and biochemistry in dogs with experimentally induced steroid hepatopathy

  M Syakalima, M Takiguchi, J Yasuda, Y Morita, A Hashimoto  VETERINARY QUARTERLY  20-  (1)  18  -22  1998/01  [Not refereed][Not invited]
   

  Digital analysis of liver ultrasound images (USGs) was compared to histological and serum enzyme activity results in dogs with steroid-induced hepatopathy. Steroid hepatopathy was used as a model for diffuse liver diseases. Prednisolone administration resulted in increased acoustic backscatter (hyperechogenicity) of the liver,vith reference to the kidney and significant depth attenuation (hyper-attenuation). Absolute changes were determined by histogram analysis of echo means (Ems) of area samples (1x1cm) of liver and kidney at the depth of 2 cm (liver-kidney contrast) and at 2 ems and 4 cm (depth attenuation). Liver-kidney contrast histograms correlated well with histology but were more sensitive than serum enzyme activity and subjective visual interpretation. Depth attenuation was the earliest detectable acoustic change. These results suggested that depth attenuation is an early and sensitive indicator of steroid hepatopathy. Liver-kidney contrast correlates well with histology and may complement biopsy examination during follow-up studies.
• Comparison of attenuation and liver-kidney contrast of liver ultrasonographs with histology and biochemistry in dogs with experimentally induced steroid hepatopathy

  M Syakalima, M Takiguchi, J Yasuda, Y Morita, A Hashimoto  VETERINARY QUARTERLY  20-  (1)  18  -22  1998/01  [Not refereed][Not invited]
   

  Digital analysis of liver ultrasound images (USGs) was compared to histological and serum enzyme activity results in dogs with steroid-induced hepatopathy. Steroid hepatopathy was used as a model for diffuse liver diseases. Prednisolone administration resulted in increased acoustic backscatter (hyperechogenicity) of the liver,vith reference to the kidney and significant depth attenuation (hyper-attenuation). Absolute changes were determined by histogram analysis of echo means (Ems) of area samples (1x1cm) of liver and kidney at the depth of 2 cm (liver-kidney contrast) and at 2 ems and 4 cm (depth attenuation). Liver-kidney contrast histograms correlated well with histology but were more sensitive than serum enzyme activity and subjective visual interpretation. Depth attenuation was the earliest detectable acoustic change. These results suggested that depth attenuation is an early and sensitive indicator of steroid hepatopathy. Liver-kidney contrast correlates well with histology and may complement biopsy examination during follow-up studies.
• Separation and quantification of corticosteroid-induced, bone and liver alkaline phosphatase isoenzymes in canine serum

  M Syakalima, M Takiguchi, J Yasuda, A Hashimoto  JOURNAL OF VETERINARY MEDICINE SERIES A-PHYSIOLOGY PATHOLOGY CLINICAL MEDICINE  44-  (9-10)  603  -610  1997/12  [Not refereed][Not invited]
   

  Quantifying alkaline phosphatase (ALP) isoenzymes in canine serum would provide a useful index in a clinical laboratory. To achieve this goal, we tested a semi-automatic assay combining wheat germ lectin (WGL) precipitation and chemical inhibition of isoenzymes of the TNS gene with levamisole to quantify bone ALP (BALP) and corticosteroid-induced ALP (CALP), respectively. The liver ALP (LALP) isoenzyme was then calculated from the equation: TALP=BALP+LALP+CALP BALP, LALP and CALP standards from serum of puppies, bile-duct ligated dogs and dogs on 4.4 mg/kg/day prednisolone for 30 days, respectively, were used. The suitability of standard sera was tested by affinity electrophoresis. Levamisole (4.2 mM) inhibits 98% of BALP and LALP but only 42% CALP. Multiplying measured CALP by 1.8 gives the total CALP value in serum. WGL precipitated 92.3% BALP, 23.3% LALP and 26.8% heated CALP standards. These values were used to adjust precipitated ALP ro obtain the exact levels of BALP. WGL was then tested on pooled serum standards in which the relative proportions of all the ALPs were known and controlled. BALP was adequately quantified except when LALP and CALP levels were extremely high. The assay was also applicable under conditions resulting in high ALP. Therefore, combining WGL and levamisole inhibition provides an adequate separation and quantification of canine ALP isoenzymes. The method has great potential for diagnostic use and should be tested further for routine implementation.
• Separation and quantification of corticosteroid-induced, bone and liver alkaline phosphatase isoenzymes in canine serum

  M Syakalima, M Takiguchi, J Yasuda, A Hashimoto  JOURNAL OF VETERINARY MEDICINE SERIES A-PHYSIOLOGY PATHOLOGY CLINICAL MEDICINE  44-  (9-10)  603  -610  1997/12  [Not refereed][Not invited]
   

  Quantifying alkaline phosphatase (ALP) isoenzymes in canine serum would provide a useful index in a clinical laboratory. To achieve this goal, we tested a semi-automatic assay combining wheat germ lectin (WGL) precipitation and chemical inhibition of isoenzymes of the TNS gene with levamisole to quantify bone ALP (BALP) and corticosteroid-induced ALP (CALP), respectively. The liver ALP (LALP) isoenzyme was then calculated from the equation: TALP=BALP+LALP+CALP BALP, LALP and CALP standards from serum of puppies, bile-duct ligated dogs and dogs on 4.4 mg/kg/day prednisolone for 30 days, respectively, were used. The suitability of standard sera was tested by affinity electrophoresis. Levamisole (4.2 mM) inhibits 98% of BALP and LALP but only 42% CALP. Multiplying measured CALP by 1.8 gives the total CALP value in serum. WGL precipitated 92.3% BALP, 23.3% LALP and 26.8% heated CALP standards. These values were used to adjust precipitated ALP ro obtain the exact levels of BALP. WGL was then tested on pooled serum standards in which the relative proportions of all the ALPs were known and controlled. BALP was adequately quantified except when LALP and CALP levels were extremely high. The assay was also applicable under conditions resulting in high ALP. Therefore, combining WGL and levamisole inhibition provides an adequate separation and quantification of canine ALP isoenzymes. The method has great potential for diagnostic use and should be tested further for routine implementation.
• イヌにおける年齢別の血清中ALP分画活性値の定量及びステロイド誘発性ALP分画活性値の診断的意義

  Syakalima Michelo, 滝口 満喜, 安田 準  The Journal of Veterinary Medical Science  59-  (10)  905  -909  1997/10
• The age dependent levels of serum ALP isoenzymes and the diagnostic significance of corticosteroid-induced ALP during long-term glucocorticoid treatment

  M Syakalima, M Takiguchi, J Yasuda, A Hashimoto  JOURNAL OF VETERINARY MEDICAL SCIENCE  59-  (10)  905  -909  1997/10  [Not refereed][Not invited]
   

  Three isoenzymes of total alkaline phosphatase (TALP) are known in canine serum: Bone alkaline phosphatase (BALP), liver alkaline phosphatase (LALP) and corticosteroid-induced alkaline phosphatase (CALF). Using an assay developed by combining selective precipitation of BALP by wheat germ lectin (WGA) and an automated levamisole inhibition method for quantifying CALF, age-related reference ranges of the isoenzymes in 75 canine serum samples were investigated. BALP comprised 96, 38 and 26% of TALP in young, middle aged and old dogs, respectively, and CALF was respectively 12, 11 and 27% of TALP. LALP was less than 10% in the young but represented more than 50% of TALP in middle aged and old dogs. Furthermore, the significance of monitoring LALP and CALF and their relationship to hepatopathy in dogs receiving long term prednisolone therapy was assessed. In this study, TALP increased in all dogs receiving prednisolone. But only LALP wits responsible in dogs with minor vacuolization of the liver, while in severely degenerated cases both LALP and CALF increased. It is concluded that a high TALP due solely to LALP, rather than LALP and CALF represents lesser liver pathologic involvement. Monitoring the 2 isoenzymes has greater significancy in assessing the level of liver damage than relying on an increased TALP value alone. Quantifying the individual isoenzymes may further be useful in assessing the clinical significance of these isoenzymes in various conditions that result in elevated TALP values.
• The age dependent levels of serum ALP isoenzymes and the diagnostic significance of corticosteroid-induced ALP during long-term glucocorticoid treatment

  M Syakalima, M Takiguchi, J Yasuda, A Hashimoto  JOURNAL OF VETERINARY MEDICAL SCIENCE  59-  (10)  905  -909  1997/10  [Not refereed][Not invited]
   

  Three isoenzymes of total alkaline phosphatase (TALP) are known in canine serum: Bone alkaline phosphatase (BALP), liver alkaline phosphatase (LALP) and corticosteroid-induced alkaline phosphatase (CALF). Using an assay developed by combining selective precipitation of BALP by wheat germ lectin (WGA) and an automated levamisole inhibition method for quantifying CALF, age-related reference ranges of the isoenzymes in 75 canine serum samples were investigated. BALP comprised 96, 38 and 26% of TALP in young, middle aged and old dogs, respectively, and CALF was respectively 12, 11 and 27% of TALP. LALP was less than 10% in the young but represented more than 50% of TALP in middle aged and old dogs. Furthermore, the significance of monitoring LALP and CALF and their relationship to hepatopathy in dogs receiving long term prednisolone therapy was assessed. In this study, TALP increased in all dogs receiving prednisolone. But only LALP wits responsible in dogs with minor vacuolization of the liver, while in severely degenerated cases both LALP and CALF increased. It is concluded that a high TALP due solely to LALP, rather than LALP and CALF represents lesser liver pathologic involvement. Monitoring the 2 isoenzymes has greater significancy in assessing the level of liver damage than relying on an increased TALP value alone. Quantifying the individual isoenzymes may further be useful in assessing the clinical significance of these isoenzymes in various conditions that result in elevated TALP values.
• Ultrasonographic appearance of orthotopic ureterocele in a dog

  M Takiguchi, J Yasuda, K Ochiai, Y Morita, A Hashimoto  VETERINARY RADIOLOGY & ULTRASOUND  38-  (5)  398  -399  1997/09  [Not refereed][Not invited]
   

  A three-year-old, intact male, Siberian husky was evaluated for a two day history of dysuria, Sonographically there was an anechoic cyst-like structure in the urinary bladder, The abnormality appeared to be a 'cyst within a cyst', which is a characteristic ultrasonographic feature of ureterocele in humans, Ultrasonography may be a useful means of establishing a diagnosis of ureterocele in dogs.
• Ultrasonographic appearance of orthotopic ureterocele in a dog

  M Takiguchi, J Yasuda, K Ochiai, Y Morita, A Hashimoto  VETERINARY RADIOLOGY & ULTRASOUND  38-  (5)  398  -399  1997/09  [Not refereed][Not invited]
   

  A three-year-old, intact male, Siberian husky was evaluated for a two day history of dysuria, Sonographically there was an anechoic cyst-like structure in the urinary bladder, The abnormality appeared to be a 'cyst within a cyst', which is a characteristic ultrasonographic feature of ureterocele in humans, Ultrasonography may be a useful means of establishing a diagnosis of ureterocele in dogs.
• Structure of the pre-early gene of canine herpesvirus.

  MIYOSHI MASAHIRO, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA, OKAZAKI KATSUNORI, KIDA HIROSHI  日本獣医学会学術集会講演要旨集  124th-  150  1997/09  [Not refereed][Not invited]
  
• MRI image observation of a lung echinococcosis rat model.

  KAWAHARA TOKIKO, ASANUMA TAKETOSHI, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA, KUWAHARA MIKINORI, MA RYO, ITO AKIRA  日本獣医学会学術集会講演要旨集  124th-  43  1997/09  [Not refereed][Not invited]
  
• Insulin stimulated secretion test by the 10% arginine hydrochloride load in the cat.

  KITAMURA TOMISHIGEKO, MORITA YUTAKA, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  124th-  199  1997/09  [Not refereed][Not invited]
  
• On the clinical significance of bovine serum adenosine deaminase activity.

  YASUDA JUN, TAKEMA SHUNSUKE, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  124th-  200  1997/09  [Not refereed][Not invited]
  
• Research on the application of endoscopic ultrasonic inspection (EUS) to small- animal clinic. (II). Hystogram analysis of the pancreas substance echo luminance of a dog having received pancreatic duct ligation.

  MORITA YUTAKA, KITAMURA TOSHIKO, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  124th-  42  1997/09  [Not refereed][Not invited]
  
• Intrahepatic lobular distribution of alkaline phosphatase (ALP) isoenzymes in the dog glucocorticoid induced hepatic disorder.

  MICHERO SHAKARIMA, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  124th-  200  1997/09  [Not refereed][Not invited]
  
• 超音波検査が有用であった犬の腸閉塞の3例

  滝口 満喜  北海道獣医師会雑誌  41-  (8)  276  -276  1997/08
• Short-term hemodialysis treatment in dogs and cats with total uretic obstruction

  T Mashita, J Yasuda, M Iijima, M Takiguchi, T Yamazaki, A Hashimoto  JAPANESE JOURNAL OF VETERINARY RESEARCH  45-  (2)  59  -65  1997/08  [Not refereed][Not invited]
   

  This study evaluated the single-pass system for the short-term dialysis treatment of dogs and cats with experimental renal failure. The hemodialyzer was equipped with a thin and highly permeable Cuprophan membrane. Four animals (two dogs and two cats) with total uretic obstruction were dialyzed twice in a one-week period. The vascular access by venipuncture of external jugular vein delivered more than 5 ml/min/kg/body weight of blood repeatedly, even for the cats. The evaluation of the effects of the blood now, dialysate flow and ultrafiltration pressure revealed that the blood now was the most important factor for effective dialysis. A 300 ml/min dialysate flow provided enough clearance of blood urea nitrogen and creatinine. The ultrafiltration pressure played an important role in ensuring that the fluid removal was constant. Laboratory studies revealed a 50.0% (range 42.0 to 59.3%) reduction of blood urea nitrogen, a 48.7% (range 42.5 to 58.7%) reduction of creatinine, and a 49.8% (range 34.3 to 66.2%) reduction of inorganic phosphate during the dialysis treatment. No dialysis disequilibrium syndrome was shown by the clinical signs. We conclude that this short-term dialysis using a single-pass system for small animals was sufficiently applicable to dogs and cats, and that the optimal duration of the dialysis was 2 hours.
• Short-term hemodialysis treatment in dogs and cats with total uretic obstruction

  T Mashita, J Yasuda, M Iijima, M Takiguchi, T Yamazaki, A Hashimoto  JAPANESE JOURNAL OF VETERINARY RESEARCH  45-  (2)  59  -65  1997/08  [Not refereed][Not invited]
   

  This study evaluated the single-pass system for the short-term dialysis treatment of dogs and cats with experimental renal failure. The hemodialyzer was equipped with a thin and highly permeable Cuprophan membrane. Four animals (two dogs and two cats) with total uretic obstruction were dialyzed twice in a one-week period. The vascular access by venipuncture of external jugular vein delivered more than 5 ml/min/kg/body weight of blood repeatedly, even for the cats. The evaluation of the effects of the blood now, dialysate flow and ultrafiltration pressure revealed that the blood now was the most important factor for effective dialysis. A 300 ml/min dialysate flow provided enough clearance of blood urea nitrogen and creatinine. The ultrafiltration pressure played an important role in ensuring that the fluid removal was constant. Laboratory studies revealed a 50.0% (range 42.0 to 59.3%) reduction of blood urea nitrogen, a 48.7% (range 42.5 to 58.7%) reduction of creatinine, and a 49.8% (range 34.3 to 66.2%) reduction of inorganic phosphate during the dialysis treatment. No dialysis disequilibrium syndrome was shown by the clinical signs. We conclude that this short-term dialysis using a single-pass system for small animals was sufficiently applicable to dogs and cats, and that the optimal duration of the dialysis was 2 hours.
• Circumstances on small animal medical care in U.S.A.

  TAKIGUCHI MITSUYOSHI  北海道獣医師会雑誌  41-  (4)  91  -94  1997/04  [Not refereed][Not invited]
  
• Study of application of ultrasonic autoscope inspection (EUS) to small animal clinic. Histogram analysis of pancreas substance echo luminance with caerulein induced pancreatitis of dog.

  MORITA YUTAKA, KITAMURA TOSHIKO, TAKIGUCHI MITSUYOSHI, YASUDA JUN, HASHIMOTO AKIRA  日本獣医学会学術集会講演要旨集  123rd-  255  1997/03  [Not refereed][Not invited]
  
• Esophageal/gastric adenocarcinoma in a dog

  M Takiguchi, J Yasuda, A Hashimoto, K Ochiai, C Itakura  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION  33-  (1)  42  -44  1997/01  [Not refereed][Not invited]
   

  A nine-year-old, intact female, mixed-breed dog was presented with a history of chronic regurgitation and weight loss. Endoscopy and contrast radiography confirmed the presence of a distal esophageal mass. The dog was euthanized, and histopathological diagnosis of the mass was an esophageal/gastric adenocarcinoma.
• Esophageal/gastric adenocarcinoma in a dog

  M Takiguchi, J Yasuda, A Hashimoto, K Ochiai, C Itakura  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION  33-  (1)  42  -44  1997/01  [Not refereed][Not invited]
   

  A nine-year-old, intact female, mixed-breed dog was presented with a history of chronic regurgitation and weight loss. Endoscopy and contrast radiography confirmed the presence of a distal esophageal mass. The dog was euthanized, and histopathological diagnosis of the mass was an esophageal/gastric adenocarcinoma.
• Ultrasonographic renal changes associated with phenylbutazone administration in three foals

  R Leveille, T Miyabayashi, SE Weisbrode, DS Biller, M Takiguchi, JF Williams  CANADIAN VETERINARY JOURNAL-REVUE VETERINAIRE CANADIENNE  37-  (4)  235  -236  1996/04  [Not refereed][Not invited]
  
• Ultrasonographic renal changes associated with phenylbutazone administration in three foals

  R Leveille, T Miyabayashi, SE Weisbrode, DS Biller, M Takiguchi, JF Williams  CANADIAN VETERINARY JOURNAL-REVUE VETERINAIRE CANADIENNE  37-  (4)  235  -236  1996/04  [Not refereed][Not invited]
  
• Ultrasonic image of liver in cat cestode infected rat.

  SAKAKIBARA YOSHIMI, TAKIGUCHI MITSUYOSHI, HASHIMOTO AKIRA, YASUDA JUN, ITO AKIRA  日本獣医学会学術集会講演要旨集  121st-  109  1996/03  [Not refereed][Not invited]
  
• Ultrasonographic Examination of Cephaloridine (Cer)-Induced Renal Lesions in Rabbits

  Katsuhiko Ishida, Akira Hashimoto, Jun Yasuda, Mitsuyoshi Takiguchi, Kunio Doi  Journal of Toxicologic Pathology  9-  (3)  273  -276  1996  [Not refereed][Not invited]
   

  Serial ultrasonographic examinations of cephaloridine (CER)-induced renal lesions were performed in rabbits. The nephrosonograms in CER-injected rabbits showed enlarged renal echoic area, increased cortical echogenicity, increased medullary hypoechoic area, and enhanced corticomedullary demarcation. The kidneys in CER-injected rabbits showed macroscopically renal enlargement, cortical discoloration, and medullary enlargement and congestion, and showed histopathologically cortical tubular necrosis, urinary casts in the distal nephrons, and congestion in the outer medulla. These results suggest that drug-induced tubular necrosis is indicated by increased renal cortical echogenicity and that ultrasonographic examination is useful for evaluation of drug-induced nephrotoxicity. © 1996, JAPANESE SOCIETY OF TOXICOLOGIC PATHOLOGY. All rights reserved.
• Ultrasonographic Examination of Cephaloridine (Cer)-Induced Renal Lesions in Rabbits

  Katsuhiko Ishida, Akira Hashimoto, Jun Yasuda, Mitsuyoshi Takiguchi, Kunio Doi  Journal of Toxicologic Pathology  9-  (3)  273  -276  1996  [Not refereed][Not invited]
   

  Serial ultrasonographic examinations of cephaloridine (CER)-induced renal lesions were performed in rabbits. The nephrosonograms in CER-injected rabbits showed enlarged renal echoic area, increased cortical echogenicity, increased medullary hypoechoic area, and enhanced corticomedullary demarcation. The kidneys in CER-injected rabbits showed macroscopically renal enlargement, cortical discoloration, and medullary enlargement and congestion, and showed histopathologically cortical tubular necrosis, urinary casts in the distal nephrons, and congestion in the outer medulla. These results suggest that drug-induced tubular necrosis is indicated by increased renal cortical echogenicity and that ultrasonographic examination is useful for evaluation of drug-induced nephrotoxicity. © 1996, JAPANESE SOCIETY OF TOXICOLOGIC PATHOLOGY. All rights reserved.
• RADIOGRAPHIC ANATOMY OF THE MEDIAL CORONOID PROCESS OF DOGS

  T MIYABAYASHI, M TAKIGUCHI, SC SCHRADER, DS BILLER  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION  31-  (2)  125  -132  1995/03  [Not refereed][Not invited]
   

  Mediolateral, flexed mediolateral, mediocaudal-laterocranial 15 degrees oblique (extended and supinated mediolateral), and craniolateral-caudomedial 20 degrees to 30 degrees oblique radiographs of 16 elbow-joint specimens were produced to study the radiographic anatomy of the medial coronoid process. On the mediolateral view, the cranial point of the coronoid process was at the level of the distal one-third of the radial epiphysis. Degree of superimposition of the proximal radius and ulna determined how the medial coronoid process was projected on the radiographs. Mediocaudal-laterocranial oblique radiographs best showed the cranial outline of the medial coronoid process with moderate superimposition of the proximal radius and ulna.
• RADIOGRAPHIC ANATOMY OF THE MEDIAL CORONOID PROCESS OF DOGS

  T MIYABAYASHI, M TAKIGUCHI, SC SCHRADER, DS BILLER  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION  31-  (2)  125  -132  1995/03  [Not refereed][Not invited]
   

  Mediolateral, flexed mediolateral, mediocaudal-laterocranial 15 degrees oblique (extended and supinated mediolateral), and craniolateral-caudomedial 20 degrees to 30 degrees oblique radiographs of 16 elbow-joint specimens were produced to study the radiographic anatomy of the medial coronoid process. On the mediolateral view, the cranial point of the coronoid process was at the level of the distal one-third of the radial epiphysis. Degree of superimposition of the proximal radius and ulna determined how the medial coronoid process was projected on the radiographs. Mediocaudal-laterocranial oblique radiographs best showed the cranial outline of the medial coronoid process with moderate superimposition of the proximal radius and ulna.
• 抗血清およびアシクロビルによる新生仔犬の犬ヘルペスウイルス感染症の発病阻止効果

  浅野真知, 滝口満喜, 安田準, 橋本晃, 喜田宏, 平井克哉  日本獣医学会学術集会講演要旨集  119th-  235  1995/03  [Not refereed][Not invited]
  
• 新しい臨床診断 キモパパインを用いた髄核融解後のMRI像に関する試験的研究

  滝口満喜, 宮林孝仁  日本獣医学会学術集会講演要旨集  118th-  65  1994/08  [Not refereed][Not invited]
  
• 臨床生化学 小動物臨床における尿中酵素の診断的意義

  松岡鎮雄, 真下忠久, 森田豊, 滝口満喜, 安田準, 山崎忠良, SYAKALIMA M, 橋本晃  日本獣医学会学術集会講演要旨集  118th-  25  1994/08  [Not refereed][Not invited]
  
• RADIOGRAPHIC APPEARANCES OF THE NASAL CONCHAE IN DOGS USING DIFFERENT SCREEN-FILM SYSTEMS - A POSTMORTEM STUDY

  T MIYABAYASHI, DS BILLER, PR HAIDER, M TAKIGUCHI  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION  30-  (4)  382  -388  1994/07  [Not refereed][Not invited]
   

  Twelve canine heads were radiographed using different intensifying screen-film combinations before and after gelatin instillation within the nasal cavity. The nasal conchal pattern was seen best in nonscreen, radiotherapy localization films. With a fast screen-film system, the conchal pattern was poorly visualized. After gelatin instillation into the nasal cavity, the conchal pattern still was visible on nonscreen films. However, the conchal pattern was more difficult to visualize on a fast screen-film system. Despite substantially increased radiographic exposures, the high diagnostic quality of nonscreen radiographs appeared to warrant their use in evaluating nasal disease. Radiographic changes on a fast screen-film system should be interpreted cautiously for destructive lesions of the nasal cavity.
• RADIOGRAPHIC APPEARANCES OF THE NASAL CONCHAE IN DOGS USING DIFFERENT SCREEN-FILM SYSTEMS - A POSTMORTEM STUDY

  T MIYABAYASHI, DS BILLER, PR HAIDER, M TAKIGUCHI  JOURNAL OF THE AMERICAN ANIMAL HOSPITAL ASSOCIATION  30-  (4)  382  -388  1994/07  [Not refereed][Not invited]
   

  Twelve canine heads were radiographed using different intensifying screen-film combinations before and after gelatin instillation within the nasal cavity. The nasal conchal pattern was seen best in nonscreen, radiotherapy localization films. With a fast screen-film system, the conchal pattern was poorly visualized. After gelatin instillation into the nasal cavity, the conchal pattern still was visible on nonscreen films. However, the conchal pattern was more difficult to visualize on a fast screen-film system. Despite substantially increased radiographic exposures, the high diagnostic quality of nonscreen radiographs appeared to warrant their use in evaluating nasal disease. Radiographic changes on a fast screen-film system should be interpreted cautiously for destructive lesions of the nasal cavity.
• Degeneration and diseases of intervertebral disk of the dog.

  TAKIGUCHI MITSUYOSHI, MIYABAYASHI TAKAHITO  小動物臨床  13-  (2)  13  -18  1994/03  [Not refereed][Not invited]
  
• SCINTIGRAPHIC AND ULTRASONOGRAPHIC DIAGNOSIS OF SOFT-TISSUE INJURY IN A THOROUGHBRED HORSE

  J WILLIAMS, T MIYABAYASHI, A RUGGLES, J YAMAMOTO, M TAKIGUCHI  JOURNAL OF VETERINARY MEDICAL SCIENCE  56-  (1)  169  -172  1994/02  [Not refereed][Not invited]
   

  A 2.5-year-old female Thoroughbred horse was referred to the veterinary teaching hospital for right front limb lameness of 1 year duration. Physical examination and diagnostic nerve blocks failed to localize the origin of the lameness. Scintigraphy with Tc-99m-MDP suggested increased radionuclide uptake in the palmar metacarpal soft tissues of the right front limb. Ultrasonographic examination revealed hypoechoic lesions in the superficial digital flexor tendon and the suspensory ligament, suggesting tendinitis and desmitis. Combined imaging modalities improved detection and characterization of the cause of a long-standing obscure lameness.
• 犬の椎間板の変性と疾患(共著)

  滝口 満喜, 宮林 孝仁  小動物臨床  13-  (2)  13  -18  1994  [Not refereed][Not invited]
  
• Scintigraphic and ultrasonographic diagnosis of soft tissue injury in a thoroughberd horse. "jointly worked"

  J. Vet. Med. Sci.  53-  (6)  1019  -1024  1994  [Not refereed][Not invited]
  
• アメリカにおける獣医免許制度について

  宮林孝仁, 滝口満喜  小動物臨床  12-  (5)  31  -33  1993/09  [Not refereed][Not invited]
  
• 犬猫の口腔内腫瘍(共著)

  滝口 満喜, 宮林 孝仁  小動物臨床  12-  (1)  11  -14  1993  [Not refereed][Not invited]
  
• Tumor in oral cavity of dog and cat.

  TAKIGUCHI MITSUYOSHI, MIYABAYASHI TAKAHITO  小動物臨床  12-  (1)  11  -14  1993/01  [Not refereed][Not invited]
  
• Acute phase reaction protein of a horse.

  OKUMURA MASAHIRO, YAMASHITA KAZUHITO, TAIRA TOMOKO, ASANO KAZUYUKI, FUKAWA YASUSHI, TAKIGUCHI MITSUYOSHI, FUJINAGA TOORU  家畜生化学研究会報  (29)  9  -18  1992/09  [Not refereed][Not invited]
  
• ウマの急性相蛋白（共著）

  家畜生化学研究会報  29: 9-18-  1992  [Not refereed][Not invited]
  
• SERUM C-REACTIVE PROTEIN (CRP) IN HORSES - THE EFFECT OF AGING, SEX, DELIVERY AND INFLAMMATIONS ON ITS CONCENTRATION

  K YAMASHITA, T FUJINAGA, M OKUMURA, M TAKIGUCHI, N TSUNODA, S MIZUNO  JOURNAL OF VETERINARY MEDICAL SCIENCE  53-  (6)  1019  -1024  1991/12  [Not refereed][Not invited]
   

  The serum concentration of C-reactive protein (CRP) in horses was measured by single radial immunodiffusion (SRID) using rabbit anti-equine CRP serum to evaluate the significance of CRP as an acute-phase reactive protein. In serum samples of clinically normal newborn thoroughbred foals before being given colostrum, serum CRP was not detectable (less-than-or-equal-to 1-mu-g/ml). The serum CRP concentration was found to increase quickly and reach a peak level of approximately 14.1-mu-g/ml in 12-month-old horses. Then, it decreased gradually to reach a low value of 5.4-mu-g/ml in the 4-year-old. In horses over 5-year-old, serum CRP concentration was stable at values of 7 to 8-mu-g/ml. In mares during the peri-natal period, serum CRP concentration decreased at 2 months before delivery, showed moderate changes just before and after delivery, and then increased to relatively high level by 2-months after delivery. The concentration of serum CRP was found to be increased at 24 hours after experimentally inflammatory stimuli in adult horses, and reached peak values of 3 to 6 times as high as the base line values at 3 to 5 days. Serum CRP concentration returned to base line values by 2 to 3 weeks after treatments. It is clear from these data that CRP is one of the acute-phase reactive proteins in horses.
• SERUM C-REACTIVE PROTEIN (CRP) IN HORSES - THE EFFECT OF AGING, SEX, DELIVERY AND INFLAMMATIONS ON ITS CONCENTRATION

  K YAMASHITA, T FUJINAGA, M OKUMURA, M TAKIGUCHI, N TSUNODA, S MIZUNO  JOURNAL OF VETERINARY MEDICAL SCIENCE  53-  (6)  1019  -1024  1991/12  [Not refereed][Not invited]
   

  The serum concentration of C-reactive protein (CRP) in horses was measured by single radial immunodiffusion (SRID) using rabbit anti-equine CRP serum to evaluate the significance of CRP as an acute-phase reactive protein. In serum samples of clinically normal newborn thoroughbred foals before being given colostrum, serum CRP was not detectable (less-than-or-equal-to 1-mu-g/ml). The serum CRP concentration was found to increase quickly and reach a peak level of approximately 14.1-mu-g/ml in 12-month-old horses. Then, it decreased gradually to reach a low value of 5.4-mu-g/ml in the 4-year-old. In horses over 5-year-old, serum CRP concentration was stable at values of 7 to 8-mu-g/ml. In mares during the peri-natal period, serum CRP concentration decreased at 2 months before delivery, showed moderate changes just before and after delivery, and then increased to relatively high level by 2-months after delivery. The concentration of serum CRP was found to be increased at 24 hours after experimentally inflammatory stimuli in adult horses, and reached peak values of 3 to 6 times as high as the base line values at 3 to 5 days. Serum CRP concentration returned to base line values by 2 to 3 weeks after treatments. It is clear from these data that CRP is one of the acute-phase reactive proteins in horses.
• 猫の外傷性口蓋破裂の外科的閉鎖法(共著)

  日本動物病院協会雑誌  22: 12-16-  1991  [Not refereed][Not invited]
  
• ISOLATION, CHARACTERIZATION, AND QUANTITATIVE-ANALYSIS OF C-REACTIVE PROTEIN FROM HORSES

  M TAKIGUCHI, T FUJINAGA, M NAIKI, S MIZUNO, K OTOMO  AMERICAN JOURNAL OF VETERINARY RESEARCH  51-  (8)  1215  -1220  1990/08  [Not refereed][Not invited]
  
• ISOLATION, CHARACTERIZATION, AND QUANTITATIVE-ANALYSIS OF C-REACTIVE PROTEIN FROM HORSES

  M TAKIGUCHI, T FUJINAGA, M NAIKI, S MIZUNO, K OTOMO  AMERICAN JOURNAL OF VETERINARY RESEARCH  51-  (8)  1215  -1220  1990/08  [Not refereed][Not invited]
  
• Massive subcutaneous hemangiosarcoma in a dog.

  FUJINAGA TORU, SUWA TAKAHIKO, NARIKIO MICHIYO, OKUMURA MASAHIRO, INOUE KUMIKO, TAKIGUCHI MITSUYOSHI, MURAKAMI MASAAKI, TAJIMA MOTOSHI, OTOMO KANJURO  日本獣医師会雑誌  42-  (7)  491  -494  1989/07  [Not refereed][Not invited]
  
• ウマの急性相蛋白に関する研究(1) ウマC-反応性蛋白の分離・精製およびその定量

  滝口 満喜  日本獣医学会学術集会講演要旨集  107回-  227  -227  1989/03
• Massive subcutaneous hemangiosarcoma in a dog.

  藤永徹, 諏訪隆彦, 成清美智代, 奥村正裕, 井上久美子, 滝口満喜, 村上正晃, 田島誉士, 大友勘十郎  日本獣医師会雑誌  42: 491-494-  (7)  1989  [Not refereed][Not invited]
  
• Masive subcutaneous hemangiosarcoma in a dog (in Jaoanese with English summary) 'jointly worked'

  J. Jpn. Vet. Assoc.  3-  491  -494  1989  [Not refereed][Not invited]
  
• Isolation, characterization, and quantitative analysis of C-reactive protein from horses. "jointly worked"

  Jap. J. Vet. Res.  37-  (2)  134  1989  [Not refereed][Not invited]
  
• Masive subcutaneous hemangiosarcoma in a dog (in Jaoanese with English summary) 'jointly worked'

  J. Jpn. Vet. Assoc.  3-  491  -494  1989  [Not refereed][Not invited]
  

Association Memberships

• 国際獣医放射線学会   日本小動物獣医学会   日本超音波医学会   日本獣医画像診断学会   日本免疫学会   日本獣医学会   

Research Projects

• 副腎皮質機能亢進症関連肝障害による肝細胞癌発症機序の解明

  日本学術振興会：科学研究費助成事業

  Date (from‐to) : 2023/04 -2026/03 

  Author : 滝口 満喜, 村上 正晃, 佐々木 東, 新坊 弦也, 横山 望
• Targetting of cancer-specific metabolic system for senolytic therapy

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)

  Date (from‐to) : 2020/04 -2025/03 

  Author : 稲波 修, 平岡 和佳子, 安井 博宣, 平田 拓, 滝口 満喜, 岡松 優子

   

  本年度はインビトロの培養細胞の系の実験で、以下の四つの結果が明らかとなった。 1）グルタミン要求性の強いヒト肺がんA549細胞やH460細胞では、放射線照射により、これらの細胞でのグルタミンの取り込みの増大とグルタメートの増大、すなわちグルタミナーゼの活性化を起こす事を明らかにした。さらに、グルタミン代謝阻害剤添加あるいはグルタミン欠乏培地下で培養すると治療線量レベルの放射線照射によって多くの老化様細胞死とSASPの培養液中への分泌を増強させることが可能である事が明らかとなった。 2）グルタミノリシス阻害条件下で放射線による老化様細胞死の誘発機構には活性酸素種の生成量、グルタチオン量の変化DNA損傷の量、さらにはDNA損傷に伴う伴うp53やp21、p16の発現変化などの既知のメカニズムの関与は殆ど無く、未知の機構の関与が示唆されている。 3）更にグルタミノリシス阻害条件下で放射線誘発する老化様細胞死を起こす条件で、Bcl-XL阻害剤ABT-737によって、この増大した老化様細胞死の多くはアポトーシスに変換できること、いわゆるセノリシスを起こせることを明らかにした。 4）一方では、同じ肺がん由来細胞でも786-o細胞ではグルタミン代謝阻害が老化様細胞の増強を起こさず、A549やH460細胞と同様の現象は観察されず、全ての肺がん細胞に於いて同様の機構が働いていない事も明らかとなった。また、脂質代謝阻害剤は老化様細胞増強には関与しなかった。 以上の結果はグルタミン要求性の強い肺がん細胞を中心にその亜致死レベルのグルタミノリシス阻害条件下で治療線量レベルの放射線照射をすると、多量の老化様細胞死の増大が起こす事を示しており、さらに、アポトーシス抑制因子の阻害剤で効率よく老化様細胞死をアポトーシスに変換できることが明確になった。
• 副腎－肝臓連関によるイヌ肝細胞癌発症メカニズムの解明

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  Date (from‐to) : 2020/04 -2023/03 

  Author : 滝口 満喜, 村上 正晃, 山崎 淳平, 池中 良徳

   

  犬の肝細胞癌における、ステロイドホルモン産生過剰を起点とする発がん仮説の検証を目指し、前年度に引き続いた検討を行った。2年目となる本年度は肝細胞癌・副腎皮質機能亢進症罹患犬における血中ステロイドホルモン・代謝産物のプロファイリング、ならびに肝臓組織でのエピゲノム変異の解析を遂行した。 ステロイドプロファイリングでは前年度に確立した測定系を発展拡大させ、新たに6種の副腎皮質ホルモンと4種の代謝産物を合わせた、合計19種類のステロイドプロファイル解析が可能となった。この新たな測定系を用いて、肝細胞癌36症例、副腎皮質機能亢進症15症例、併発11症例、コントロール19症例を対象として、血液中のステロイドプロファイルを解析した。しかし、血液中のステロイドプロファイルに疾患特異性、もしくは疾患群間での差が認められなかった。 DNAメチル化の解析では、実験的ステロイドホルモン誘発性脂肪肝、副腎皮質腫機能亢進症併発肝細胞癌および肝細胞癌単独症例の腫瘍組織と腫瘍近傍組織の、合計3種類の肝臓組織を用い、多段階発癌仮説の検証を試みた。その結果、脂肪肝・近傍組織・腫瘍組織と段階的なメチル化変化が存在し、腫瘍においてメチル化レベルの高い遺伝子が6、低い遺伝子が12、抽出できた。 現時点では血中ステロイドプロファイルに肝細胞癌と副腎皮質機能亢進症との関連を示唆する血中ステロイドプロファイリングは見つかっていない。一方で、ステロイド肝から腫瘍への段階的なDNAメチル化レベルの変化が存在することが明らかになった。犬の肝細胞癌においてステロイドが発症に役割を果たしていることは十分に予想され、本年度の成果を複合的かつ詳細に解析することで、多段階発がん説の検証が進む。
• Sono-mechanotherapy for neurite outgrowth

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Research (Exploratory)

  Date (from‐to) : 2018/06 -2020/03 

  Author : Mitsuyoshi Takiguchi

   

  Axon regeneration is crucial for the functional recovery after spinal cord injury. Myelin-related growth inhibitors, such as Nogo-A, inhibit neurites regrowth and elongation. Low-intensity pulsed ultrasound (LIPUS) is known to show neuroprotective effects in neurodegenerative diseases. In the present study, we evaluated the feasibility of LIPUS for enhancing axon regeneration in the presence of Nogo-A. Low-intensity pulsed ultrasound was exposed to rat cortical neurons in vitro after Nogo-A treatment. The total length of neurites per cell in LIPUS group was significantly larger than that of non-LIPUS group. There was no significant difference in the number of neurites. The results suggest that LIPUS may enhance the elongation of neurites in the presence of Nogo-A while may not affect on neurites sprouting and branching. This preliminary study implies the feasibility of LIPUS for axonal regeneration.
• Development of a novel therapy by targeting for cancer specific glutamine and acetate metabolism

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2017/04 -2020/03 

  Author : Inanami Osamu

   

  In this study, in order to clarify a novel metabolic target of radiotherapy for cancer, we established a mitochondrial-dependent metabolic assay for cancer metabolism using electron spin resonance (ESR) with a new oxygen probe. Using this method, it was clarified that ATP production by not only glucose metabolism but also electron transport system (ETC) was increased by irradiation as a survival strategy of solid tumors. Furthermore, we have found that this radiation-induced energy response can induce strong radiosensitization by glutaminolysis involved in the TCA cycle and mitochondrial inhibitory drugs. In particular, it was demonstrated that glutamine synthase inhibitors and ETC inhibitors can cause a strong radiosensitizing effect.
• Novel cancer therapy for spontaneous animal tumor using ultrasound irradiation with microbubble

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2014/04 -2017/03 

  Author : TAKIGUCHI MITSUYOSHI, KUDO Nobuki

   

  This study revealed that sonoporation, the combination of ultrasound and microbubbles, enhances the antitumor effect of a chemotherapeutic agent with minimum invasiveness in preclinical canine thyroid and bladder cancer models. Moreover, the results of this study clarified the mechanism of sonoporation. This preclinical study may contribute to establish sonoporation as a theranostic tool in cancer therapy.
• The development of a novel sensitizing radiation therapy in conjunction with microbubbles and ultrasound exposure

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2014/04 -2016/03 

  Author : TAKIGUCHI Mitsuyoshi, SASAKI Noboru

   

  The purpose of this study is to clarify the sensitizing effect of irradiation-induced apoptosis after ultrasound exposure in the presence of microbubbles. Mouse sequemous cell carcinoma cell was irradiated right after the ultrasonic exposure with microbubbles resulted in sensitizing effect of 1.1. After considering optimal irradiation timing in the light of the effect of cell cycle variation of ultrasound exposure, it was suggested that 9 hours after the ultrasound exposure was suitable. However, the sensitizing effect of irradiation-induced apoptosis remained 1.1. Further study is needed using 3-dimensional cell culture system.
• 動物自然発生腫瘍に対するマイクロバブル併用超音波照射による実用的な新規がん治療

  文部科学省：科学研究費補助金(基盤研究(B))

  Date (from‐to) : 2014 -2016 

  Author : 滝口 満喜
• The role of Babesia gibsoni heat shock protein 90 on the proliferation and the drug resistance.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2012/04 -2015/03 

  Author : YAMASAKI Masahiro, OHTA Hiroshi, TAKIGUCHI Mitsuyoshi

   

  In this study, Heat shock protein 90 (Hsp90) in Babesia gibsoni, a blood protozoa in dogs, was analyzed. The Babesia gibsoni heat shock protein 90 (BgHsp90) gene was cloned and sequenced. Moreover, anti recombinant BgHsp90 antibody was developed. Additionally, the expression of BgHsp90 was observed. As results, it was shown that BgHsp90 might contribute the proliferation of the parasites, and that the expression of BgHsp90 would be elevated when the parasites were exposed to the high temperature. Meanwhile, Hsp90 inhibitors could suppress the proliferation of B. gibsoni in vitro. This result suggested that the BgHsp90 should be one of the drug targets for the treatment of canine babesiosis.
• Molecular mechanisms of pathogenesis and parasite persistence in canine visceral leishmaniasis

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2012/04 -2015/03 

  Author : KATAKURA Ken, KATO Hirotomo, TAKIGUCHI Mitsuyoshi, ATO Manabu

   

  Visceral leishmaniasis (VL) in humans in the Indian subcontinent is caused by infection with Leishmania donovani and the transmission is thought to be anthroponotic. The prevalence of L. donovani infection and the role of VL transmission in dogs have not been well investigated. The present epidemiological study showed that approximately 10% of stray dogs were serological or PCR positive in the blood in a VL endemic area in Bangladesh, but no severe clinical symptoms were observed. Experimental infection of beagle dogs with L. donovani showed that no remarkable changes were detected in clinical sings, hematological and serum biochemical values, and peripheral T lymphocyte subpopulations. Although these dogs were negative for PCR examinations, antibodies were detected during the experimental period. Taken together, the results suggest that dogs infected with L. donovani maintain the latent infection, but serve as the reservoir animal for the transmission of human VL.
• マイクロバブルと超音波を利用した新しい増感放射線療法の開発

  文部科学省：科学研究費補助金(挑戦的萌芽研究)

  Date (from‐to) : 2014 -2015 

  Author : 滝口 満喜
• バベシア原虫の増殖および薬剤耐性獲得における熱ショックタンパク質９０の役割の解明

  文部科学省：科学研究費補助金(基盤研究(C))

  Date (from‐to) : 2012 -2014 

  Author : 山崎 真大, 大田 寛, 滝口 満喜

   

  犬に感染する赤血球内寄生原虫であるBabesia gibsoniのheat shock protein 90 (hsp90)について、平成25年度は以下の研究を行った。平成24年度には、組み替えhsp90蛋白質の作成のためにプラスミドを作成したが、これを大腸菌内に取り込ませ、大腸菌を用いた組み替えBabesia gibsoni hsp90 (Bghsp90)の発現を実施し、組み替えBghsp90蛋白質の作成に成功した。大腸菌の大量培養を実施することでこれを大量に発現させ、組み替え蛋白質に結合させたHisタグを利用して組み替えBghsp90蛋白質を精製した。さらに、組み替えBghsp90蛋白質をウサギに免疫し、ウサギの体内でこの蛋白質に対する抗血清を作成、さらに抗血清から組み替えBghsp90蛋白質を用いてアフィニティー精製を行ってウサギ抗組み替えBghsp90ポリクローナル抗体を作成した。この抗体がB. gibsoniのhsp90に特異的に結合することを確認した後に、これを用いてウェスタンブロッティングおよび免疫染色を行い、B. gibsoniの持つBghsp90蛋白質を検出、定量することに成功した。そこで、これらの技術を用いて、B. gibsoniの野生株、ならびに抗原虫薬であるジミナゼン製剤に耐性を持つ耐性株のhsp90量を比較し、耐性株ではhsp90発現量が減少していることを明らかにした。この結果は、昨年実施したリアルタイムPCR法を用いた遺伝子発現量の定量を行った実験とよく相関したことから、B. gibsoniのジミナゼン耐性株ではBghsp90の発現量が低下していることが明らかになった。
• Establishment of novel and objective ultrasonographic examination and its application to clinical veterinary medicine

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2011 -2013 

  Author : TAKIGUCHI MITSUYOSHI, YAMASAKI Masahiro, OHTA Hiroshi

   

  To establish novel and objective ultrasonographic examination and apply it to clinical veterinary medicine, fundamental parameters of normal canine pancreatic perfusion were determined by using contrast agent Sonazoid. Then, novel diagnostic critetria for acute pancreatitis focused on pancreatic perfusion changes was established based on analysis of contrast-enhanced ultrasonographic examination in experimentally induced acute pancreatitis model in the dog. In addition, left atrial phasic function in healthy dogs by means of 2-D speckle tracking echocardiography with time-left atrial area curve analysis was evaluated. The repeatability and reproducibility of obtained measurements were adequate. This results indicates that studies are needed to determine clinical applicability in canine patients.
• 客観的病態診断を可能とする新たな超音波検査法の確立と獣医臨床応用の進展

  文部科学省：科学研究費補助金(基盤研究(C))

  Date (from‐to) : 2011 -2013 

  Author : 滝口 満喜, 山崎 真大, 大田 寛

   

  本研究の目的は、伴侶動物における超音波検査による客観的病態診断の確立に向けた基盤を形成し、その臨床応用の進展を検討することである。まず、超音波造影剤ソナゾイドを用いた造影超音波検査の臨床応用の適用拡大を目的として、未だに診断が困難な膵炎に着目し、正常犬の膵臓の灌流状態に関する基礎的知見を得た。次いで、実験的に作出したセルレイン誘発性急性膵炎モデルにおける造影超音波検査所見を解析して、膵臓の血流変化に着目した急性膵炎の新しい診断基準を確立することを目指した。その結果、超音波造影剤ソナゾイドのボーラス投与(0.01ml/kg)では膵臓の至適観察時間が16秒と短かったため、ソナゾイドを0.05ml/kgで持続点滴することで観察時間を150秒に大幅に延長させる造影検査法を確立した(J Vet Med Sci, 75:1601-1607, 2013) 。また、セルレイン誘発浮腫性膵炎では、正常時に比べて膵実質のエコー輝度のピークまでの到達時間が短縮し、最高輝度が上昇することが明らかとなった(J Vet Intern Med, 28:496-503, 2014) 。これまでソナゾイドを用いた犬の膵臓の造影超音波検査に関する報告は皆無であり、本研究成果によって、犬の膵臓の造影超音波検査法が確立され、実験的に作出した浮腫性膵炎における膵臓の灌流状態の変化の特徴が明らかになったことは、現在進行中の膵炎の臨床例での解析に向けてその礎を築くことができた。臨床例での解析結果も実験膵炎と同様の結果が得られ、本年6月のアメリカ獣医内科学会で発表する予定である。加えて、左心房機能に着目した心エコー図検査による新たな心機能評価法の確立を目指し、正常犬の左心房面積の時相による変化を解析して正常値を確立するとともに、本手法の再現性を明らかにした(Am J Vet Res, 74:864-869)。
• Gut microbiota and gut mucosal barrier in hibernating animals

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2011 -2012 

  Author : SONOYAMA Kei, TSUBOTA Toshio, TAKIGUCHI Mitsuyoshi

   

  This study investigated the control of gut microbiota and gut mucosal barrier in hibernating animals. In hibernating Syrian hamsters, we observed higher concentrations of secretory antibodies in the small intestinal contents, suggesting its contribution to protection against infection. Indeed, no bacteria were observed in the mesenteric lymph node of hibernating hamsters. Analysis of lymphocyte profiles in the immune tissues indicated that ability to protect against infection and to regulate immune function may be maintained in the gut of hibernating hamsters. Furthermore, we found that cell renewal of intestinal epithelium is markedly retarded in hibernating hamsters.
• Gut microbiota and gut mucosal barrier in hibernating animals

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(挑戦的萌芽研究)

  Date (from‐to) : 2011 -2012 

  Author : Kei SONOYAMA, Toshio TSUBOTA, Mitsuyoshi TAKIGUCHI

   

  This study investigated the control of gut microbiota and gut mucosal barrier in hibernating animals. In hibernating Syrian hamsters, we observed higher concentrations of secretory antibodies in the small intestinal contents, suggesting its contribution to protection against infection. Indeed, no bacteria were observed in the mesenteric lymph node of hibernating hamsters. Analysis of lymphocyte profiles in the immune tissues indicated that ability to protect against infection and to regulate immune function may be maintained in the gut of hibernating hamsters. Furthermore, we found that cell renewal of intestinal epithelium is markedly retarded in hibernating hamsters.
• 膜透過性ペプチド融合アポトーシス誘導タンパク質を用いた新規放射線がん治療

  日本学術振興会：科学研究費助成事業 挑戦的萌芽研究

  Date (from‐to) : 2009 -2010 

  Author : 稲波 修, 滝口 満喜, 山盛 徹

   

  本研究では、p34^-サイクリンB1によるリン酸化部位である34番目のスレオニン(T)をアラニン(A)へ置換したT34A変異サバイビンのN末端に膜透過性ペプチドTAT配列を付加した融合蛋白質を作成し、腫瘍治療におけるTATペプチドによる蛋白質導入技術の有効性と、T34A変異サバイビンのcolon26マウス移植固形腫瘍モデルへの局所投与と放射線照射の併用による抗腫瘍効果について検討した。TATペプチドを付加したサバイビン蛋白質は、腫瘍組織への直接投与で、短時間に腫瘍細胞内へ導入された。X線2Gyの単独照射は、有意な腫瘍の増殖遅延を示さなかった。また、TAT融合T34A変異サバイビンの腫瘍細胞への単独投与も、有意な増殖遅延を示さなかった。一方、X線2GyとTAT融合T34A変異サバイビンの併用は顕著な増殖遅延を示した。組織学的な検索によりX線2GyとTAT融合T34A変異サバイビンの併用による抗腫瘍効果はアポトーシスの誘導によるものであることが示された。さらに、併用により腫瘍組織中の増殖細胞数の減少が観察されたことより、アポトーシス誘導は増殖細胞に特異的に起こることが示唆された。細胞周期関連蛋白質の検出から放射線照射によるG2/M期停止が示唆された。これらの結果から、TAT融合T34A変異サバイビンとX線2Gyの併用による抗腫瘍効果はX線照射で生じる細胞周期チェックポイントに関連したT34A変異サバイビンによるアポトーシス誘導の増強であると考えられた。以上の結果から、TATペプチドによる蛋白質導入技術が腫瘍治療において有効であり、T34A変異サバイビンの投与は腫瘍の放射線治療において、放射線による抗腫瘍作用を増強することが明らかにされた。
• 膜透過性ペプチド融合アポトーシス誘導タンパク質を用いた新規放射線がん治療

  文部科学省：科学研究費補助金(挑戦的萌芽研究)

  Date (from‐to) : 2009 -2010 

  Author : 稲波 修, 滝口 満喜, 山盛 徹

   

  本研究では、p34^-サイクリンB1によるリン酸化部位である34番目のスレオニン(T)をアラニン(A)へ置換したT34A変異サバイビンのN末端に膜透過性ペプチドTAT配列を付加した融合蛋白質を作成し、腫瘍治療におけるTATペプチドによる蛋白質導入技術の有効性と、T34A変異サバイビンのcolon26マウス移植固形腫瘍モデルへの局所投与と放射線照射の併用による抗腫瘍効果について検討した。TATペプチドを付加したサバイビン蛋白質は、腫瘍組織への直接投与で、短時間に腫瘍細胞内へ導入された。X線2Gyの単独照射は、有意な腫瘍の増殖遅延を示さなかった。また、TAT融合T34A変異サバイビンの腫瘍細胞への単独投与も、有意な増殖遅延を示さなかった。一方、X線2GyとTAT融合T34A変異サバイビンの併用は顕著な増殖遅延を示した。組織学的な検索によりX線2GyとTAT融合T34A変異サバイビンの併用による抗腫瘍効果はアポトーシスの誘導によるものであることが示された。さらに、併用により腫瘍組織中の増殖細胞数の減少が観察されたことより、アポトーシス誘導は増殖細胞に特異的に起こることが示唆された。細胞周期関連蛋白質の検出から放射線照射によるG2/M期停止が示唆された。これらの結果から、TAT融合T34A変異サバイビンとX線2Gyの併用による抗腫瘍効果はX線照射で生じる細胞周期チェックポイントに関連したT34A変異サバイビンによるアポトーシス誘導の増強であると考えられた。以上の結果から、TATペプチドによる蛋白質導入技術が腫瘍治療において有効であり、T34A変異サバイビンの投与は腫瘍の放射線治療において、放射線による抗腫瘍作用を増強することが明らかにされた。
• Genes regulating B cell domionant infiltration into autoimmune sialitis lesions and their intergenic interaction

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2006 -2008 

  Author : TAKIGUCHI Mltsuyoshi

   

  シェーグレン症候群(Sjogren's syndrome ; SS)モデルIQIマウスにおけるB細胞優勢の下顎腺炎(B細胞性下顎腺炎)を遺伝学的に解析した。その結果、B細胞性下顎腺炎の量的形質遺伝子座(QTL)は少なくとも3個有り、単独で効果を発揮する主要QTL(autoimmune sialitis in IQImice, associated locus 1 ; Asq1)が第6染色体前端に、Asq1の効果を増強するエピスタシスQTLが第3染色体後方(Asq2)に、また、逆にAsq1の効果を抑制する抵抗性遺伝子座が第1染色体後方(Asq3)に位置することを明らかになった。Asq1の1LOD区間内に位置するIca1に注目し、そのcDNA塩基配列を対照C57BL/6(B6)と比較した結果、IQI-B6間で6個の一塩基多型があり、2個のアミノ酸多型が生じていた。また、ウエスタンブロット法では、IQエマウスの血清中に抗Ica1自己抗体の存在を示すバンドカミ検出された。以上の結果より、IQIマウスにおいて、Ica1に対するB細胞自己免疫が成立していることが明らかになり、原因として、1ca1の塩基多型の可能性が挙げられた。
• Novel molecular pathogenesis of autoimmune diseases : Self attack as a result of breakdown of central tolerance

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2005 -2006 

  Author : TAKIGUCHI Mitsuyoshi, TANIYAMA Hiroyuki, KONNO Akihiro

   

  The ability to discriminate between self and non-self antigens is pivotal for the immune system to defense the body specifically invading microorganisms. The breakdown of mechanisms to tolerance self-antigens can result in the pathological autoimmunity with the destruction or disruption of the body's own tissues by the immune system. We demonstrated that IQI/Jic mice, an animal model for Sjogren's syndrome, spontaneously develop inflammatory lesions mainly infiltrated with CD4^+ T cells and B cells not only in the lacrimal and salivary glands but also in multiple organs including the lung, pancreas, and kidney at advanced ages. We next identified Kallikrein-13 was an autoantigen associated with systemic autoimmunity in IQI/Jic mice. In addition, we found that IL-2 production of T cells was impaired at the transcriptional level in IQI/Jic mice. It was suggested that defective production of IL-2 resulting from poor activation of T cells could abrogate the self-toleration mechanism through IL-2 and create the basis of autoimmune disease in IQI/Jic mice. Finally, we performed thymectomy on day 3 after birth (D3Tx) in IQI/Jic mice, which is known to eliminate regulatory T cells (Treg) to address whether there is a loss of toleration mechanism through Treg and it contributes to the early development of autoimmune lesions in IQI/Jic mice. It was suggested that spontaneous autoimmune lesions in IQI/Jic mice develop independently of the self-toleration mechanism through Treg. In conclusion, in IQI/Jic mice, dysfunction of activated T cells to produce optimal levels of IL-2 could abrogate the self-toleration mechanisms not through Treg and cause persistent activation of T cells reactive with autoantigens. Moreover, autoimmunity against autoantigens including Klk-13, might be crucial in the etiology of disease progression from salivary gland-specific to systemic disorder. These findings obtained from this work could contribute to a better understanding of the pathogenesis of Sjogren's syndrome in humans and other autoimmune diseases.
• Novel molecular pathogenesis of autoimmune diseases : Self attack as a result of breakdown of central tolerance

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 2005 -2006 

  Author : Mitsuyoshi TAKIGUCHI, Hiroyuki TANIYAMA, Akihiro KONNO

   

  The ability to discriminate between self and non-self antigens is pivotal for the immune system to defense the body specifically invading microorganisms. The breakdown of mechanisms to tolerance self-antigens can result in the pathological autoimmunity with the destruction or disruption of the body's own tissues by the immune system. We demonstrated that IQI/Jic mice, an animal model for Sjogren's syndrome, spontaneously develop inflammatory lesions mainly infiltrated with CD4^+ T cells and B cells not only in the lacrimal and salivary glands but also in multiple organs including the lung, pancreas, and kidney at advanced ages. We next identified Kallikrein-13 was an autoantigen associated with systemic autoimmunity in IQI/Jic mice. In addition, we found that IL-2 production of T cells was impaired at the transcriptional level in IQI/Jic mice. It was suggested that defective production of IL-2 resulting from poor activation of T cells could abrogate the self-toleration mechanism through IL-2 and create the basis of autoimmune disease in IQI/Jic mice. Finally, we performed thymectomy on day 3 after birth (D3Tx) in IQI/Jic mice, which is known to eliminate regulatory T cells (Treg) to address whether there is a loss of toleration mechanism through Treg and it contributes to the early development of autoimmune lesions in IQI/Jic mice. It was suggested that spontaneous autoimmune lesions in IQI/Jic mice develop independently of the self-toleration mechanism through Treg. In conclusion, in IQI/Jic mice, dysfunction of activated T cells to produce optimal levels of IL-2 could abrogate the self-toleration mechanisms not through Treg and cause persistent activation of T cells reactive with autoantigens. Moreover, autoimmunity against autoantigens including Klk-13, might be crucial in the etiology of disease progression from salivary gland-specific to systemic disorder. These findings obtained from this work could contribute to a better understanding of the pathogenesis of Sjogren's syndrome in humans and other autoimmune diseases.
• 特定領域研究「動物疾患ジェノミクス」新規創設を目指す企画と調査

  日本学術振興会：科学研究費助成事業 基盤研究(C)

  Date (from‐to) : 2004 -2004 

  Author : 稲葉 睦, 小川 博之, 国枝 哲夫, 鈴木 勝士, 北川 均, 滝口 満喜

   

  1)学術雑誌等の調査により、過去40年間に国内外で報告された動物の遺伝性疾患、ならびに動物のゲノム解析について、その内容と現在の研究動向を検討した。その概要は、獣医学教科書である「獣医内科学」等に記載のほか、獣医臨床遺伝研究会(平成17年3月)、獣医学会学術集会シンポジウム(平成17年4月)、ならびにHP(平成17年5月開設予定)にて報告する。 2)総括班、調整班が4回の討議を行い、上記を含め以下の合意に達した。 ◆対象とする疾患:特に国内で問題となっている、あるいは特徴的な疾患で、その解析が、産業動物の損耗防止や分子育種につながる牛・豚の疾患、人間社会との共存や福祉の向上に資する犬・猫の疾患、生命科学や基礎医学の基礎理論構築、医療への応用に発展し得る動物疾患。 ◆研究の展開:(1)特定の単一遺伝子疾患の原因遺伝子異常の解析、ならびに発症の分子機構とそれに基づく分子の生理機能の解明。(2)多因子疾患臨床症例の詳細な病態分類と原因となる分子・遺伝子異常の探索。 ◆研究組織:特定領域研究の創設に際しては、臨床知見の集積を含めた研究内容の特性に鑑みて5年間程度の研究展開を目指す。総括班、調整班のもとに上記(1)と(2)の研究展開の基盤となる2〜3の計画研究班を設置して重点的に基礎研究を実施するほか、2年間の公募研究をそれぞれ20件程度ずつ採用する。 2)下記の研究集会、公開シンポジウムを開催し、動物遺伝性疾患概念の周知と研究者-臨床獣医師間交流等を図った。 ◆「近未来の小動物医療-分子医学の潮流-」平成16年9月(札幌)Dr.U.Giger (Univ.Pennsylvania)ら講師4名、参加者約200名。 ◆「臨床遺伝研究会・農水事業・科研費ジョイントフォーラム」平成17年3月(天童)講演者延べ25名、参加者約80名。 これらをもとに、特定領域研究等の具体化を進める。
• 特定領域研究「動物疾患ジェノミクス」新規創設を目指す企画と調査

  文部科学省：科学研究費補助金(基盤研究(C))

  Date (from‐to) : 2004 -2004 

  Author : 稲葉 睦, 小川 博之, 国枝 哲夫, 鈴木 勝士, 北川 均, 滝口 満喜

   

  1)学術雑誌等の調査により、過去40年間に国内外で報告された動物の遺伝性疾患、ならびに動物のゲノム解析について、その内容と現在の研究動向を検討した。その概要は、獣医学教科書である「獣医内科学」等に記載のほか、獣医臨床遺伝研究会(平成17年3月)、獣医学会学術集会シンポジウム(平成17年4月)、ならびにHP(平成17年5月開設予定)にて報告する。2)総括班、調整班が4回の討議を行い、上記を含め以下の合意に達した。◆対象とする疾患:特に国内で問題となっている、あるいは特徴的な疾患で、その解析が、産業動物の損耗防止や分子育種につながる牛・豚の疾患、人間社会との共存や福祉の向上に資する犬・猫の疾患、生命科学や基礎医学の基礎理論構築、医療への応用に発展し得る動物疾患。◆研究の展開:(1)特定の単一遺伝子疾患の原因遺伝子異常の解析、ならびに発症の分子機構とそれに基づく分子の生理機能の解明。(2)多因子疾患臨床症例の詳細な病態分類と原因となる分子・遺伝子異常の探索。◆研究組織:特定領域研究の創設に際しては、臨床知見の集積を含めた研究内容の特性に鑑みて5年間程度の研究展開を目指す。総括班、調整班のもとに上記(1)と(2)の研究展開の基盤となる2〜3の計画研究班を設置して重点的に基礎研究を実施するほか、2年間の公募研究をそれぞれ20件程度ずつ採用する。2)下記の研究集会、公開シンポジウムを開催し、動物遺伝性疾患概念の周知と研究者-臨床獣医師間交流等を図った。◆「近未来の小動物医療-分子医学の潮流-」平成16年9月(札幌)Dr.U.Giger (Univ.Pennsylvania)ら講師4名、参加者約200名。◆「臨床遺伝研究会・農水事業・科研費ジョイントフォーラム」平成17年3月(天童)講演者延べ25名、参加者約80名。これらをもとに、特定領域研究等の具体化を進める。
• 組織の破壊と低形成-細胞間ジャンクションの分子構築破綻と疾患の遺伝的基盤-

  日本学術振興会：科学研究費助成事業 萌芽研究

  Date (from‐to) : 2003 -2004 

  Author : 滝口 満喜, 稲葉 睦, 田島 誉士, 高桑 雄一

   

  本研究は、上皮や実質組織の破壊や低形成をともなう多彩な動物疾患の臨床知見の集積と分子病態解析の成果から派生したアイディアに基づくものである。本計画は、"細胞間接合の遺伝的異常と疾患"という新しい観点から、従来、遺伝性素因の存在を疑われながら病因のとらえどころがなかった組織破壊を伴う疾患や臓器低形成の分子機構を解明することを将来目標としている。本萌芽研究では、上記仮説実証の第一歩として、タイトジャンクション(TJ)に分布するCL分子ファミリーに焦点を絞り、まず正常組織における分子群の臓器、組織分布を明らかにし、次いで腎、肝、消化管、皮膚の組織破壊、あるいは低形成を呈する臨床例におけるCL分子の異常を検証することを目的とした。クローディン(CL)分子ファミリーは現在までにヒト、マウスでいずれもCL-1〜CL-18までの18分子種が知られている。代表者らは、これまで牛CL-16に関する解析を行ってきたが、本研究計画では小動物(犬と猫)を対象とするため、まずこれら18種のCLを各々同定するためのツールとして特異抗体を作製することから着手した。犬、猫、個々のCL分子にそれぞれ対応する抗体を作製するのは煩雑であり、また交差性が期待されるため、まず犬CLsに対する抗体を作出、必要に応じて抗猫CLs抗体を作製することとした。抗原として用いる各CL分子C末端ペプチド(C末端の細胞質ドメイン30-40アミノ酸残基)を調製するために、各CL分子のcDNAをRACE法で単離し、GST-融合タンパク質ベクターにクローン化した。cDNA単離に利用する臓器、あるいはプライマーはヒトでの知見に基づいた。GST-融合タンパク質として得た各分子から必要なC末端ペプチドを精製し、ウサギに免疫して、アフィニティー精製によって特異抗体を得た。次いで、イムノブロッティング、ならびに正常組織を用いた蛍光抗体法で反応性と特異性を検定した。臨床例を用いた検討では、様々な疾患でCL分子の局在や発現量に変化が認められたが、現時点では疾患特異的に変動する分子の特定には至らず、今後、さらなる解析が必要である。
• 組織の破壊と低形成-細胞間ジャンクションの分子構築破綻と疾患の遺伝的基盤-

  文部科学省：科学研究費補助金(萌芽研究)

  Date (from‐to) : 2003 -2004 

  Author : 滝口 満喜, 高桑 雄一, 田島 誉士, 稲葉 睦

   

  本研究は、上皮や実質組織の破壊や低形成をともなう多彩な動物疾患の臨床知見の集積と分子病態解析の成果から派生したアイディアに基づくものである。本計画は、"細胞間接合の遺伝的異常と疾患"という新しい観点から、従来、遺伝性素因の存在を疑われながら病因のとらえどころがなかった組織破壊を伴う疾患や臓器低形成の分子機構を解明することを将来目標としている。本萌芽研究では、上記仮説実証の第一歩として、タイトジャンクション(TJ)に分布するCL分子ファミリーに焦点を絞り、まず正常組織における分子群の臓器、組織分布を明らかにし、次いで腎、肝、消化管、皮膚の組織破壊、あるいは低形成を呈する臨床例におけるCL分子の異常を検証することを目的とした。クローディン(CL)分子ファミリーは現在までにヒト、マウスでいずれもCL-1〜CL-18までの18分子種が知られている。代表者らは、これまで牛CL-16に関する解析を行ってきたが、本研究計画では小動物(犬と猫)を対象とするため、まずこれら18種のCLを各々同定するためのツールとして特異抗体を作製することから着手した。犬、猫、個々のCL分子にそれぞれ対応する抗体を作製するのは煩雑であり、また交差性が期待されるため、まず犬CLsに対する抗体を作出、必要に応じて抗猫CLs抗体を作製することとした。抗原として用いる各CL分子C末端ペプチド(C末端の細胞質ドメイン30-40アミノ酸残基)を調製するために、各CL分子のcDNAをRACE法で単離し、GST-融合タンパク質ベクターにクローン化した。cDNA単離に利用する臓器、あるいはプライマーはヒトでの知見に基づいた。GST-融合タンパク質として得た各分子から必要なC末端ペプチドを精製し、ウサギに免疫して、アフィニティー精製によって特異抗体を得た。次いで、イムノブロッティング、ならびに正常組織を用いた蛍光抗体法で反応性と特異性を検定した。臨床例を用いた検討では、様々な疾患でCL分子の局在や発現量に変化が認められたが、現時点では疾患特異的に変動する分子の特定には至らず、今後、さらなる解析が必要である。
• A clinical study on tumor dormancy therapy by anti-angiogenesis and duff erentiation agents administered to dogs with advanced spontaneous tumors.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2002 -2004 

  Author : KADOSAWA Tsuyoshi, TKIGUCHI Mitsuyoshi, OCHIAI Kenji, KASAOKA Tatsuhiko, OOIZUMI Iwao, KUSAKA Masami

   

  A clinical evaluation of the novel angiogenesis inhibitor TNP 470, Thalidomide and differentiation agent OCT (22-oxa-calcitriol) for tumor dormancy was performed. Dogs with advanced spontaneous malignant tumors and their owners' consent to a pilot study were eligible TNP 470, a dose of 12.5-223mg/m^2., was administered to 47 dogs subcutaneously or intravenously 1-3 times per week. Lameness or shivering were encountered in 9 dogs. Seizure was shown in 4 dogs, and bleeding, anorexia and vomiting were in a dog, respectively. Thalidomide, daily oral dose of 5.0-20mg/kg B.W., was administered to 23 dogs. Lameness, sedation or softer feces were observed in 6 dogs. Though there was a tendency to increase of these clinical signs in higher doses, most were induced by a sleeping effect in itself. OCT, daily oral dose of 0.03-0.12 Lt g/kg B.W. was administered to 31 dogs. In 21 dogs receiving over 2 weeks, 7 dogs showed hypercalcemia. Hypercalcemia resolved, followed by a 7-day rest period. In each drugs, side effects and changes in blood chemical analysis were not serious. Evaluations of anti-tumor effects, such as inhibition of tumor growth and metastasis, were difficult without a control group. But primary or metastatic lesions shrank by these agents in a few dogs. A small number of dogs with highly metastasizing tumors such as osteosarcoma survived for a long time without progressive disease. In this clinical study, dogs with advanced malignant tumors tolerated well to TNP-470, Thalidomide and OCT, and some encouraging anti-tumor effects were shown. Further studies of tumor dormancy therapy by anti-angiogenesis and differentiation agents are warranted.
• A clinical study on tumor dormancy therapy by anti-angiogenesis and duff erentiation agents administered to dogs with advanced spontaneous tumors.

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2002 -2004 

  Author : Tsuyoshi KADOSAWA, 日下 雅実, 笠岡 達彦, Kenji OCHIAI, Mitsuyoshi TKIGUCHI, 大泉 巌雄, 日下 雅美, Tatsuhiko KASAOKA, Iwao OOIZUMI, Masami KUSAKA

   

  A clinical evaluation of the novel angiogenesis inhibitor TNP 470, Thalidomide and differentiation agent OCT (22-oxa-calcitriol) for tumor dormancy was performed. Dogs with advanced spontaneous malignant tumors and their owners' consent to a pilot study were eligibleTNP 470, a dose of 12.5-223mg/m^2., was administered to 47 dogs subcutaneously or intravenously 1-3 times per week. Lameness or shivering were encountered in 9 dogs. Seizure was shown in 4 dogs, and bleeding, anorexia and vomiting were in a dog, respectively. Thalidomide, daily oral dose of 5.0-20mg/kg B.W., was administered to 23 dogs. Lameness, sedation or softer feces were observed in 6 dogs. Though there was a tendency to increase of these clinical signs in higher doses, most were induced by a sleeping effect in itself. OCT, daily oral dose of 0.03-0.12 Lt g/kg B.W. was administered to 31 dogs. In 21 dogs receiving over 2 weeks, 7 dogs showed hypercalcemia. Hypercalcemia resolved, followed by a 7-day rest period. In each drugs, side effects and changes in blood chemical analysis were not serious.Evaluations of anti-tumor effects, such as inhibition of tumor growth and metastasis, were difficult without a control group. But primary or metastatic lesions shrank by these agents in a few dogs. A small number of dogs with highly metastasizing tumors such as osteosarcoma survived for a long time without progressive disease.In this clinical study, dogs with advanced malignant tumors tolerated well to TNP-470, Thalidomide and OCT, and some encouraging anti-tumor effects were shown. Further studies of tumor dormancy therapy by anti-angiogenesis and differentiation agents are warranted.
• Molecular mechanisms for the assembly of the red cell membrane skeleton during erythroid cell development

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2002 -2003 

  Author : INABA Mutsumi, TAKAKUWA Yuichi, SAITO Masayuki, MAEDA Yoshimitsu, INANAMI Osamu, YAMAMOTO Masayuki

   

  The "unit-assembly hypothesis" for red cell membrane skeleton during erythroid development was investigated. Stable transfectants of major constituents of red cell transmembrane proteins were developed in K562 and HEK293 cells using retroviral vectors. Band 3 and glycophorinCweretransportedtotheplasmamembranevia the ER/Golgi pathway. Band 3-binding domain of ankyrin (AnkN90) and protein 4.1 showed membranous localization in the cytoplasm with co-localization with markers for the ER and the Golgi apparatus in band 3-transfeted cells despite of the difference in erythroid differentiation. Endogenous membrane skeletal proteins including ankyrin, protein 4.1, and spectrin also exhibited membranous distribution. These results indicate that the assembly of membrane skeletal proteins occurs at an early stage of erythroidal development on the surface of the ER but not on the inner surface of the plasma membrane. Stable expression of band 3 caused a reduction in a proliferation rate of K562 cells to approximately 70% that of non-transfected cells. Reduced production of hemoglobin suggestive of erythroid differentiation was also observed. However, no significant changes in intracellular localization of skeletal proteins and cell morphology was detected Synthesis and assembly to the ER of skeletal proteins involving spectrin and subsequent synthesis and localization to the plasma membrane of band 3 were observed in erythroid precursor cells from peripheral blood of cattle in two phase liquid culture system, indicating that the assembly of membrane skeletal proteins to the ER occurs in physiolosical erythropoiesis. The present study demonstrated that the assembly of the red cell membrane skeleton is an event which takes place at early stages of erythroid development on the ER. The exact mechanism by which a small unit of the ekeleton formed on intracellular vesicles are integrated in the plasma membrane remained to be clarified.
• 膜タンパク質のポストゴルジ/ERレトリーバル小胞輸送における新規分子機構の構築

  日本学術振興会：科学研究費助成事業 萌芽研究

  Date (from‐to) : 2002 -2003 

  Author : 稲葉 睦, 高桑 雄一, 滝口 満喜

   

  本研究の目的は、牛グライコフォリンC(GP-C)の発現、遺伝子型-表現型連関について従来得た以下の仮説を検証し、S85によるポストゴルジ膜移行、あるいはERレトリーバル制御の分子機構を解明することを目的とする。 <仮説1>S85のリン酸化等の翻訳後修飾がポストゴルジ膜移行を決定づける未知のシグナルとして機能している。あるいは、<仮説2>S85を含む周辺配列が未知の膜移行認識シグナルとなっている。仮説1との相乗作用もあり得る。 1.仮説1を検証するために、GST-fused GP-C(C末端細胞質ドメイン)を調製し、これを基質とするProtein kinaseを解析した。その結果、Casein kinase I (CKI)がGP-CのC-末端細胞質ドメインをリン酸化することが判明した。CKIは、遺伝子多型であるS85、N85のGP-Cの両者をリン酸化することから、S85以外のSer/Thr残基もリン酸化されることが推定された。 2.COS細胞、K562細胞におけるGP-C発現系を構築し、それらのリン酸化動態と細胞膜への移行を解析した。これらの細胞ではGP-Cのリン酸化が観察されたが、少なくともCKI阻害剤存在下でのリン酸化の程度は対照と、あるいはGP-CS85とN85とで相違なく、また細胞膜移行の程度にも顕著な差は認められなかった。 3.翻訳後修飾としてO-GlcNAc付加を推定し、その検証を行ったが、O-GlcNAc存在は検出されなかった。 4.仮説2を検証するためにS85を中心にC末端細胞質ドメインのアミノ酸残基を欠く変異体を作製し、培養細胞に導入してその膜移行を解析したところ、むしろにC末端領域を欠く変異体の膜移行が低下することが判明した。これら変異体は、ERに留まり分解除去を受けた。
• 膜タンパク質のポストゴルジ/ERレトリーバル小胞輸送における新規分子機構の構築

  文部科学省：科学研究費補助金(萌芽研究)

  Date (from‐to) : 2002 -2003 

  Author : 稲葉 睦, 高桑 雄一, 滝口 満喜

   

  本研究の目的は、牛グライコフォリンC(GP-C)の発現、遺伝子型-表現型連関について従来得た以下の仮説を検証し、S85によるポストゴルジ膜移行、あるいはERレトリーバル制御の分子機構を解明することを目的とする。<仮説1>S85のリン酸化等の翻訳後修飾がポストゴルジ膜移行を決定づける未知のシグナルとして機能している。あるいは、<仮説2>S85を含む周辺配列が未知の膜移行認識シグナルとなっている。仮説1との相乗作用もあり得る。1.仮説1を検証するために、GST-fused GP-C(C末端細胞質ドメイン)を調製し、これを基質とするProtein kinaseを解析した。その結果、Casein kinase I (CKI)がGP-CのC-末端細胞質ドメインをリン酸化することが判明した。CKIは、遺伝子多型であるS85、N85のGP-Cの両者をリン酸化することから、S85以外のSer/Thr残基もリン酸化されることが推定された。2.COS細胞、K562細胞におけるGP-C発現系を構築し、それらのリン酸化動態と細胞膜への移行を解析した。これらの細胞ではGP-Cのリン酸化が観察されたが、少なくともCKI阻害剤存在下でのリン酸化の程度は対照と、あるいはGP-CS85とN85とで相違なく、また細胞膜移行の程度にも顕著な差は認められなかった。3.翻訳後修飾としてO-GlcNAc付加を推定し、その検証を行ったが、O-GlcNAc存在は検出されなかった。4.仮説2を検証するためにS85を中心にC末端細胞質ドメインのアミノ酸残基を欠く変異体を作製し、培養細胞に導入してその膜移行を解析したところ、むしろにC末端領域を欠く変異体の膜移行が低下することが判明した。これら変異体は、ERに留まり分解除去を受けた。
• Molecular mechanisms for the assembly of the red cell membrane skeleton during erythroid cell development

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2002 -2003 

  Author : Mutsumi INABA, 滝口 満喜, Osamu INANAMI, Yoshimitsu MAEDA, Masayuki YAMAMOTO, Yuichi TAKAKUWA, Masayuki SAITO

   

  The "unit-assembly hypothesis" for red cell membrane skeleton during erythroid development was investigated.Stable transfectants of major constituents of red cell transmembrane proteins were developed in K562 and HEK293 cells using retroviral vectors. Band 3 and glycophorinCweretransportedtotheplasmamembranevia the ER/Golgi pathway. Band 3-binding domain of ankyrin (AnkN90) and protein 4.1 showed membranous localization in the cytoplasm with co-localization with markers for the ER and the Golgi apparatus in band 3-transfeted cells despite of the difference in erythroid differentiation. Endogenous membrane skeletal proteins including ankyrin, protein 4.1, and spectrin also exhibited membranous distribution. These results indicate that the assembly of membrane skeletal proteins occurs at an early stage of erythroidal development on the surface of the ER but not on the inner surface of the plasma membrane.Stable expression of band 3 caused a reduction in a proliferation rate of K562 cells to approximately 70% that of non-transfected cells. Reduced production of hemoglobin suggestive of erythroid differentiation was also observed. However, no significant changes in intracellular localization of skeletal proteins and cell morphology was detected Synthesis and assembly to the ER of skeletal proteins involving spectrin and subsequent synthesis and localization to the plasma membrane of band 3 were observed in erythroid precursor cells from peripheral blood of cattle in two phase liquid culture system, indicating that the assembly of membrane skeletal proteins to the ER occurs in physiolosical erythropoiesis.The present study demonstrated that the assembly of the red cell membrane skeleton is an event which takes place at early stages of erythroid development on the ER. The exact mechanism by which a small unit of the ekeleton formed on intracellular vesicles are integrated in the plasma membrane remained to be clarified.
• Development of antitumor drugs to enhance apoptotic cell death and clinical application

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2000 -2001 

  Author : INANAMI Osamu, TAKIGUCHI Mitsuyoshi, MATSUDA Akira, KUWABARA Mikinori

   

  Clinically, the combination treatment of solid tumor cells with an anticancer drug and radiation was widely used to enhance cell killing. 1-(3-C-Ethynyl-β-D-rijbo-pentofuranosyl) cytosine (ECyd) was newly developed as an anticancer drug to induce cell death by inhibiting RNA synthesis. In this study, we examined whether the exposure of human gastric adenocarcinoma MKN45 cells to X rays in the presence of ECyd at the limited concentration ranges with no apoptosis induced apoptotic cell death. MKN45 cells were treated with 0.1μM ECyd for 1 h before irradiation. After irradiation with 20 Gy, apoptotic as well as swelling cells were morphologically discriminated by fluorescence microscopy combined with propidium iodide staining or electron microscopy and were scored. When cells were treated with ECyd alone, only 5% of either apoptotic or necrotic cells appeared. When cells were exposed to X rays alone, about 5% of apoptotic and about 45% of swelling cells appeared. However, when cells were exposed to X rays in the presence of 0.1μM ECyd, about 25% of totali cells was apoptotic cells and about 10% of total cells was swelling cells. Apoptosis induced by treating cells with ECyd and X irradiation was significantly reduced by the treatment with Ac-DEVD-CHO (caspase 3 inhibitor) or TPCK (chymotrypsin-like protease inhibitor). These results suggested that caspase 3 and chymotrypsin-like proteases were responsible for apoptosis induced by co-treatment with ECyd and X rays. In this study, it was demonstrated that co-treatment of MKN45 cells with ECyd and X rays activated G2-phase-linked apoptotic signaling associated with caspase 3 and chymotrypsin like protease. These data may provide useful information to develop clinical treatment of radiation combined with anticancer drugs for solid tumors.
• Development of antitumor drugs to enhance apoptotic cell death and clinical application

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 2000 -2001 

  Author : Osamu INANAMI, Mitsuyoshi TAKIGUCHI, Akira MATSUDA, Mikinori KUWABARA

   

  Clinically, the combination treatment of solid tumor cells with an anticancer drug and radiation was widely used to enhance cell killing. 1-(3-C-Ethynyl-β-D-rijbo-pentofuranosyl) cytosine (ECyd) was newly developed as an anticancer drug to induce cell death by inhibiting RNA synthesis. In this study, we examined whether the exposure of human gastric adenocarcinoma MKN45 cells to X rays in the presence of ECyd at the limited concentration ranges with no apoptosis induced apoptotic cell death. MKN45 cells were treated with 0.1μM ECyd for 1 h before irradiation. After irradiation with 20 Gy, apoptotic as well as swelling cells were morphologically discriminated by fluorescence microscopy combined with propidium iodide staining or electron microscopy and were scored. When cells were treated with ECyd alone, only 5% of either apoptotic or necrotic cells appeared. When cells were exposed to X rays alone, about 5% of apoptotic and about 45% of swelling cells appeared. However, when cells were exposed to X rays in the presence of 0.1μM ECyd, about 25% of totali cells was apoptotic cells and about 10% of total cells was swelling cells. Apoptosis induced by treating cells with ECyd and X irradiation was significantly reduced by the treatment with Ac-DEVD-CHO (caspase 3 inhibitor) or TPCK (chymotrypsin-like protease inhibitor). These results suggested that caspase 3 and chymotrypsin-like proteases were responsible for apoptosis induced by co-treatment with ECyd and X rays. In this study, it was demonstrated that co-treatment of MKN45 cells with ECyd and X rays activated G2-phase-linked apoptotic signaling associated with caspase 3 and chymotrypsin like protease. These data may provide useful information to develop clinical treatment of radiation combined with anticancer drugs for solid tumors.
• MRL/lprマウスにおける肺病変発症機序の解明

  日本学術振興会：科学研究費助成事業 奨励研究(A)

  Date (from‐to) : 1999 -2000 

  Author : 滝口 満喜

   

  本研究の目的は、MRL/lprマウスにおけるCD28/CTLA4-B7補助シグナル非依存性の肺病変について、遺伝背景の異なるマウスを用いることで病変形成に関わる原因遺伝子を探索すること、さらに、肺病変発症機序を解明するために、免疫組織化学的および臨床病理学的に詳細に解析して、病変に関わるサイトカインを同定し、抗サイトカイン抗体の投与実験によってその関わりを証明することである。 まず、MRL/lprに加えて、遺伝背景の異なるC57BL6/lprおよびC3H/He/lprマウスにもCTLA4IgGを生後24時間以内から1回100μg、週3回腹腔内に投与し、20週齢で肺の病理組織学的検索を行った。その結果、MRL/lprではCTLA4IgG投与の有無に関わらず強い肺病変が認められた。C57BL6/lprではCTLA4IgG投与により肺病変が有意に抑制され、C3H/He/lprでは対照と同程度の軽度の単核球浸潤が認められた。以上のことから、CD28/CTLA4-B7補助シグナル非依存性に生じる肺病変は、MRL遺伝背景とともにlpr遺伝子つまり、Fas分子の異常によって誘導されることが示唆された。 次に、MRL/lprマウスの肺病変発症機序の解明を目的として、MRL/lprマウスの肺組織から単核球を分離し、フローサイトメトリーによる解析を行った。表面抗原の解析から、CTLA4IgG投与によってT細胞の活性化は抑制されたが、マクロファージの活性化は抑制されないことが判明した。また、細胞内サイトカインの解析から、マクロファージからのインターフェロンガンマの産生が抑制されないことが明らかとなった。以上の事から、MRL/lprマウスにおけるCD28/CTLA4-B7補助シグナル非依存性に生じる肺病変の形成には、マクロファージから産生されるインターフェロンガンマが関わっていることが示唆された。
• MRL/lprマウスにおける肺病変発症機序の解明

  文部科学省：科学研究費補助金(奨励研究(A))

  Date (from‐to) : 1999 -2000 

  Author : 滝口 満喜

   

  本研究の目的は、MRL/lprマウスにおけるCD28/CTLA4-B7補助シグナル非依存性の肺病変について、遺伝背景の異なるマウスを用いることで病変形成に関わる原因遺伝子を探索すること、さらに、肺病変発症機序を解明するために、免疫組織化学的および臨床病理学的に詳細に解析して、病変に関わるサイトカインを同定し、抗サイトカイン抗体の投与実験によってその関わりを証明することである。まず、MRL/lprに加えて、遺伝背景の異なるC57BL6/lprおよびC3H/He/lprマウスにもCTLA4IgGを生後24時間以内から1回100μg、週3回腹腔内に投与し、20週齢で肺の病理組織学的検索を行った。その結果、MRL/lprではCTLA4IgG投与の有無に関わらず強い肺病変が認められた。C57BL6/lprではCTLA4IgG投与により肺病変が有意に抑制され、C3H/He/lprでは対照と同程度の軽度の単核球浸潤が認められた。以上のことから、CD28/CTLA4-B7補助シグナル非依存性に生じる肺病変は、MRL遺伝背景とともにlpr遺伝子つまり、Fas分子の異常によって誘導されることが示唆された。次に、MRL/lprマウスの肺病変発症機序の解明を目的として、MRL/lprマウスの肺組織から単核球を分離し、フローサイトメトリーによる解析を行った。表面抗原の解析から、CTLA4IgG投与によってT細胞の活性化は抑制されたが、マクロファージの活性化は抑制されないことが判明した。また、細胞内サイトカインの解析から、マクロファージからのインターフェロンガンマの産生が抑制されないことが明らかとなった。以上の事から、MRL/lprマウスにおけるCD28/CTLA4-B7補助シグナル非依存性に生じる肺病変の形成には、マクロファージから産生されるインターフェロンガンマが関わっていることが示唆された。
• STUDY ON CLINICAL EVALUATION OF SERUM AND URINE ENZYMES IN ORDER TO IMPROVE ACCURACYIN VETERINARYMEDICINE

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 1997 -1998 

  Author : YASUDA Jun, TAKIGUCHI Mitsuyoshi

   

  Serum alkaline phosphates (ALP) and adenosine deaminase (ADA) were evaluated in the cases of liver and tumor diseases in cows and dogs by using an auto analyzer. We had established a quantification method for that ALP isoenzymes in the serum combined with white germ lectin precipitation and chemical inhibition of bone ALP (BALP) and liver ALP (LALP) isoenzymes with levamisole to quantify BALP and corticosteroid-induced ALP (CALP), respectively. Because CALP and BALP were known, LALP was calculated from the equation : ALP=BALP+LALP+CALP.We discussed with the relationship between serum ALP isoenzymes and the degree of glucocorticoid induced hepatopathy. The changes in ALP, LALP and CALP were compared to changes in vacuolization of the hepatic parenchyma CALP elevation was found to directly correlate with the severity of vacuolization unlike LALP that increased in all cases treated with glucocorticoids and was non-specific to the degree of vacuolization. Bovine serum and liver tissues which were diagnosed as being from abnormal livers were investigated to determine the relationship between the serum ADA and the degree of mononuclear cell infiltration in the liver. The serum ADA was elevated accompanied by an increase of inflammatory cell infiltration in the liver. Thus, serum ADA analysis of cows might be a predictive index of inflammatory change in the liver. It was suggested that the ADA activity in the canine serum is insatable in the condition of preservation. Serum ADA in cases of canine lymphoma, leukeamia, liver tumor and meningioma increased before chemotherapy. The ADA levels tended to decrease after chemotherapy. Its fluctuation has different clinical significance to that of ALP or lactate dehydrogenase (LDH) in the cases of tumor disorders. We suggest that ADA analysis in dogs may provide one predictive index for tumor behavior.
• STUDY ON CLINICAL EVALUATION OF SERUM AND URINE ENZYMES IN ORDER TO IMPROVE ACCURACYIN VETERINARYMEDICINE

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 1997 -1998 

  Author : Jun YASUDA, Mitsuyoshi TAKIGUCHI

   

  Serum alkaline phosphates (ALP) and adenosine deaminase (ADA) were evaluated in the cases of liver and tumor diseases in cows and dogs by using an auto analyzer. We had established a quantification method for that ALP isoenzymes in the serum combined with white germ lectin precipitation and chemical inhibition of bone ALP (BALP) and liver ALP (LALP) isoenzymes with levamisole to quantify BALP and corticosteroid-induced ALP (CALP), respectively. Because CALP and BALP were known, LALP was calculated from the equation : ALP=BALP+LALP+CALP.We discussed with the relationship between serum ALP isoenzymes and the degree of glucocorticoid induced hepatopathy. The changes in ALP, LALP and CALP were compared to changes in vacuolization of the hepatic parenchyma CALP elevation was found to directly correlate with the severity of vacuolization unlike LALP that increased in all cases treated with glucocorticoids and was non-specific to the degree of vacuolization. Bovine serum and liver tissues which were diagnosed as being from abnormal livers were investigated to determine the relationship between the serum ADA and the degree of mononuclear cell infiltration in the liver. The serum ADA was elevated accompanied by an increase of inflammatory cell infiltration in the liver. Thus, serum ADA analysis of cows might be a predictive index of inflammatory change in the liver. It was suggested that the ADA activity in the canine serum is insatable in the condition of preservation. Serum ADA in cases of canine lymphoma, leukeamia, liver tumor and meningioma increased before chemotherapy. The ADA levels tended to decrease after chemotherapy. Its fluctuation has different clinical significance to that of ALP or lactate dehydrogenase (LDH) in the cases of tumor disorders. We suggest that ADA analysis in dogs may provide one predictive index for tumor behavior.
• 自己免疫疾患の分子機構

  Date (from‐to) : 1998
• Molecular Mechanism of Autoimmune disease

  Date (from‐to) : 1998
• 慢性腎疾患の進行に及ぼす摂取蛋白量の影響

  日本学術振興会：科学研究費助成事業 奨励研究(A)

  Date (from‐to) : 1995 -1995 

  Author : 滝口 満喜

   

  本研究の目的は、外科的に作出した慢性腎不全の猫に対する摂取蛋白量の影響を光学および電子顕微鏡を用いて組織学的に検索するとともに、慢性腎不全の進展因子として注目されている各種サイトカインや成長因子の関わりを検証することであった。 慢性腎不全猫の作出に当たっては、右腎摘出に加え左腎動脈を部分結紮する腎亜全摘モデルの作出を試みたが、高度の手術手技を要することに加えて術後の状態が不安定であったことから、猫に与えるストレスが過大すぎると判断し、腎毒性を有する薬物(ピューロマイシン・アミノヌクレオシド)を投与することで腎障害の誘発を試みた。このピューロマイシン・アミノヌクレオシドはラットに特異的に腎障害を引き起こすことが知られているが、猫での報告はない。そのため、投与量および投与間隔を変えて実験を行う必要があった。その結果、ピューロマイシン・アミノヌクレオシドを猫に投与することによって蛋白尿をきたし、一部の糸球体に硬化病変が誘発されることが示唆された。また、電子顕微鏡によって糸球体上皮細胞足突起の癒合が観察された。血清生化学的にはBUNおよびクレアチニン値に上昇が認められた。尿中酵素ではNAGに変化が認められ、尿中サイトカインではIL-6が病変の進行に伴い増加する傾向が伺えた。 以上のことからピューロマイシン・アミノヌクレオシドは猫に腎障害をもたらすことが示唆された。このモデルにおける病変の進展に及ぼす摂取蛋白量の影響については、現在、実験を継続中である。
• 慢性腎疾患の進行に及ぼす摂取蛋白量の影響

  文部科学省：科学研究費補助金(奨励研究(A))

  Date (from‐to) : 1995 -1995 

  Author : 滝口 満喜

   

  本研究の目的は、外科的に作出した慢性腎不全の猫に対する摂取蛋白量の影響を光学および電子顕微鏡を用いて組織学的に検索するとともに、慢性腎不全の進展因子として注目されている各種サイトカインや成長因子の関わりを検証することであった。慢性腎不全猫の作出に当たっては、右腎摘出に加え左腎動脈を部分結紮する腎亜全摘モデルの作出を試みたが、高度の手術手技を要することに加えて術後の状態が不安定であったことから、猫に与えるストレスが過大すぎると判断し、腎毒性を有する薬物(ピューロマイシン・アミノヌクレオシド)を投与することで腎障害の誘発を試みた。このピューロマイシン・アミノヌクレオシドはラットに特異的に腎障害を引き起こすことが知られているが、猫での報告はない。そのため、投与量および投与間隔を変えて実験を行う必要があった。その結果、ピューロマイシン・アミノヌクレオシドを猫に投与することによって蛋白尿をきたし、一部の糸球体に硬化病変が誘発されることが示唆された。また、電子顕微鏡によって糸球体上皮細胞足突起の癒合が観察された。血清生化学的にはBUNおよびクレアチニン値に上昇が認められた。尿中酵素ではNAGに変化が認められ、尿中サイトカインではIL-6が病変の進行に伴い増加する傾向が伺えた。以上のことからピューロマイシン・アミノヌクレオシドは猫に腎障害をもたらすことが示唆された。このモデルにおける病変の進展に及ぼす摂取蛋白量の影響については、現在、実験を継続中である。
• Studies on developmental and manifold applications of diagnostic ultrasound.

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(一般研究(B))

  Date (from‐to) : 1994 -1995 

  Author : Akira HASHIMOTO, Mitsuyoshi TAKIGUCHI, Jun YASUDA

   

  Results obtained from the present research can be summarized as follows :1. Clinical applications of endoscopic ultrasonography (EUS) in small animal medicine.EUS examinations were done on abdominal viscera in normal dogs and on Caerulein-induced acute pancreatitis and hematoma and cystic lesions in the gastric wall experimentally prepared. It was suggested that EUS was highly useful to image the digestive tract, gall bladder and pancreas. The results obtained from experimental study, suggest that EUS provides useful information especially on pancreatic disease and focal lesions in the digestive tract. Therefore, it is concluded that EUS is an effective diagnostic procedure for small animal practice.2. Application of ultrasound-guided biopsy techniques.On the experimental studies of steroid-induced hepatopathy, Puromycin-induced renal damage and experimental fatty liver in cat and pathobiological changes of the prostate for collection of prostatic fluid, ultrasound-guided biopsy was done for collection of the liver, kidney and prostatic specimens, respectively. From the results of these experiments, it is demonstrated that ultrasound-guided biopsy is useful, safety and accurate procedure for collection of tissue specimens in small animal medicine.3. Application of the US for the research procedure of experimental animals.The results of serial US examination of Cephaloridine-induced renal lesions suggest that US examination is useful for evaluation of drug-induced nephrotoxicity. The results of US examination of parasitic cysts of the liver infected with Taenia taeniaeformis suggest that US examination is useful for the diagnosis of rat liver parasitic cysts like hydatid as well as following-up development of metacestodes in the liver. These results strongly suggest that US investigation is useful as a effective analytical procedure of the research of experimental animals.4. Application of ultrasound in diagnosis of neoplasia of small animals.Ultrasonographic findings were obtained from various kind of tumor by US examination. Usefulness of color-doppler ultrasound and ultrasound-guided biopsy are also evaluated for detailed and definite diagnosis of the neoplasia.As a conclusion, the results of the present research may become useful information for development of veterinary diagnostic imaging.
• 小動物の超音波診断

  Date (from‐to) : 1993
• 小動物自然発生疾病の画像診断

  Date (from‐to) : 1993
• Small Animal Dignostic Ultrasound

  Date (from‐to) : 1993
• Small animal Diagnostic Imaging

  Date (from‐to) : 1993
• 自己免疫の分子機構
• 自然発生動物疾病の画像診断
• Molecular pathogenesis of autoimmune diseases
• Diagnostic imaging of spontanous animal diseases