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Partial Purification and Characterization of the Rat Parotid Gland Protein Kinase Catalyzing Phosphorylation of Matured Destrin at Ser-2

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Advances in Enzyme Research 2014年第2期2卷 100-112页

作者： Eriko Osumi Chihiro Kondo Mitsumasa Mizuno Takahiro Suzuki Mamoru Matsubara kazuo shimozato Takao Kanamori Department of Biochemistry School of Dentistry Aichi-Gakuin University Nagoya Japan Department of Bioscience and Biotechnology Faculty of Bioenvironmental Science Kyotogakuen University Kyoto Japan Department of Maxillofacial Surgery School of Dentistry Aichi-Gakuin University Nagoya Japan

Destrin, also called actin-depolymerizing factor (ADF), exists in resting parotid tissue as phosphorylated (inactive) and dephosphorylated (active) forms, and β-adrenergic stimulation of this tissue induces dephosphorylation of destrin. It is suggested that destrin dephosphorylation is involved in cortical F-actin disruption observed in parallel with β-agonist-induced amylase secretion. At present, the phosphorylation/dephosphorylation mechanism of destrin in parotid tissue is not known. We previously detected, in a crude rat parotid extract, a constitutively active protein kinase catalyzing phosphorylation of destrin;however, its identification has been hampered by difficulty in its enrichment. The purpose of this study was to explore a simple purification method(s) for this enzyme. To this end, we first developed a high-throughput dot-blot assay for the kinase with an anti-phosphodestrin antibody and then studied its purification by column chromatography on several media. We found that the kinase could be partially purified by sequential chromatography on DEAE-cellulose, phenyl-Sepharose, and hydroxyapatite columns. In each chromatography, however, the kinase could be eluted, at the cost of resolution, only by sharp increases in the elution power of the eluent;gradual increases in the elution power resulted in unacceptably poor recovery. We confirmed that enzymatic properties of the kinase were not basically altered during the purification. Further purification of the kinase was achieved by native polyacrylamide gel electrophoresis (PAGE), which resolved the kinase activity into two bands and separated the activity from most proteins (the kinase activity after PAGE was detected with destrin-coated polyvinylidene difluoride membranes and the anti-phosphodestrin antibody). The two bands seem to constitute the major destrin-phosphorylating activity in the resting rat parotid gland. We here report its partial purification and characterization together with the detecti

关键词： Protein Kinase Purification Protein Kinase Assay Destrin Kinase Actin-Depolymerizing Factor Rat Parotid Gland

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Serum vitamin A, C, E and carotenoid levels and risk of malignant transformation among oral leukoplakia： a prospective study

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中国口腔颌面外科杂志 2008年第B5期6卷 121-122页

作者： Toru Nagao Saman Warnakulasuriya Hideo Fukano Koji Suzuki Shuji Hashimoto kazuo shimozato 不详 Department of Oral and Maxillofacial Surgery Okazaki City Hospital. Okazaki 444-8553 Japan Department of Oral Medicine WHO Collaborating Centre for Oral Cancer/Precancer King's College Dental Institute at Guy's King's & St. Thomas' Hospitals. London UK Department of Maxillofacial Surgery Aichi-Gakuin University School of Dentistry. Nagoya 464-8651 Japan Department of Public Health Fujita Health University School of Health Sciences Department of Hygiene Fujita Health University School of Medicine Toyoake 470-1192 Japan

Purpose: To evaluate any association between serum micronutrients levels and the risk of malignant transformation of oral leukoplakia。Methods: A prospective hospital based study on pathologically diagnosed oral leukoplakia (n=52) was undertaken during 1996 to 2004。Serum vitamin A (retinol), C, E (a -tocopherol) and carotenoid levels were measured at baseline by the HPLC method。Those who were lost to follow up (minimum 12 months) or did not complete blood analysis were excluded。Multivariate adjusted relative risks (RR) for malignant transformation of oral leukoplakia were calculated with the Cox proportional hazard model。Results: 39 patients with leukoplakias (median age 67, range 35-80) were evaluated。During follow-up (median 52 months:range 12-78 months), 7 carcinomas (2 invasive and 1 micro-invasive, 1 verrucous carcinoma and 3 ca in-situ) were detected。Median serum beta-carotene levels in the group with malignant changes was 20.7μg/dl compared with 44.9μg/dl in those that did not transform (P=0.001)。Having a higher serum beta-carotene level had a significantly inverse association with malignant transformation by the Cox proportional hazard model (RR= 0.08, 95%CI 0.01-0.99, P=-0.049)。Vitamin A, C and E were not significant。Conelusiom A high serum beta-carotene level was associated with reduced risk of malignant transformation of oral leukoplakia, and a result was consistent with previous epidemiological findings of dietary protection of beta-carotene for oral cancer。Further evidence for the preventive role of micronutrients for potentially malignant disorders of the oral cavity should be explored。

关键词：口腔黏膜白斑病治疗方法血清维生素A 类胡萝卜素

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Functional reconstruction of maxillary oropharyngeal defect- non-surgical, prosthetic rehabilitation of articulation

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中国口腔颌面外科杂志 2008年第B5期6卷 46-46页

作者： shimozato kazuo Department of Maxillofacial Surgery School of Dentistry A ichi-Gakuin University Nagoya Japan

The prognosis of the maxillary or oropharyngeal cancer patients is now rapidly improving by the progress of surgical techniques itself, also, by multi modal treatment with chemo-, radio-therapy.

关键词：上颌骨口腔癌治疗方法外科手术

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Dose Distributions in Simulated Electron Radiotherapy with Intraoral Cones Using Treatment Planning System

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International Journal of Medical Physics, Clinical Engineering and Radiation Oncology 2017年第3期6卷 280-289页

作者： Tomohiro shimozato Kuniyasu Okudaira Department of Radiological Technology Gifu University of Medical Science Gifu Japan Department of Radiotherapy Nagoya University Hospital Aichi Japan

Aim: This study aims to evaluate the difference between depth data from an intraoral cone and a conventional irradiation tube calculated using a treatment planning system (TPS), and that measured using an intraoral cone for electron radiotherapy. Background: A TPS is only compatible with conventional irradiation tubes. However, such systems are not suitable for determining dose distributions when a special cone is employed. Materials and Methods: Dose distributions were calculated using the beam data for mounted intraoral cones using a TPS. Then, the dose distribution by field size was calculated for a low-melting-point lead alloy using the beam data for a mounted conventional tube. The calculated data were evaluated against the measured intraoral-cone depth data based on the dose and depth differences. Results: The calculated data for the intraoral cone case did not match the measured data. However, the depth data obtained considering the field size determined for the lead alloy using the conventional tube were close to the measured values for the intraoral cone case. The difference in the depth at which the absorbed dose was 50% of the maximum value of the percentage depth dose was less than ±4 mm for the generalized Gaussian pencil beam convolution algorithm and less than ±1 mm for the electron Monte Carlo algorithm. Conclusion: It was found that the measured and calculated dose distributions were in agreement, especially when then electron Monte Carlo algorithm was used. Thus, the TPS can be employed to determine dose distributions for intraoral cone applications.

关键词： Treatment Planning System Electron Radiotherapy Intraoral Cone Depth Dose Algorithm

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Direct Measurement of Medical Linear Accelerator Electron Beam Width at Scattering Foil Position

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International Journal of Medical Physics, Clinical Engineering and Radiation Oncology 2017年第1期6卷 1-7页

作者： Tomohiro shimozato Yuichi Aoyama Department of Radiological Technology School of Health Science Gifu University of Medical Science Gifu Japan Department of Radiotherapy Kobe University Hospital Kobe Japan

Purpose: The aim of this study was to develop a method for the direct measurement of electron beam width and distribution at the scattering foil on the carrousel in a medical linear accelerator gantry head, which differs from an existing indirect method for measuring the focal spot size using a camera or metallic slit located outside the gantry head. Methods: The electron beam emitted by the linear accelerator was used to irradiate radiochromic film mounted on the scattering foil on the carrousel, which was not used for clinical treatment. The electron beam width at the scattering foil position was then evaluated using the full width at half maximum of the Gaussian distribution approximated from each one dimensional distribution of the irradiated radiochromic film. Results: The electron beam width at the scattering foil position was found to be 3.1 to 6.4 mm in the crossline direction and 2.8 to 5.5 mm in the inline direction with electron energy of 4 to 16 MeV. The two-dimensional distribution of the electron beam was therefore elliptical or distorted in shape, not circular. Conclusions: Direct measurement of the electron beam width at the scattering foil in the carrousel of a medical linear accelerator is possible, though the use of lower sensitivity film in addition to indirect methods is expected to bring about better results. However, as this method does not allow for direct measurement of the incident angle of the accelerated electron beam, further improvements and refinements are still needed.

关键词： Medical Linear Accelerator Electron Beam Distribution Radiochromic Film Scattering Foil

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ADAMTS13 and von Willebrand factor are useful biomarkers for sorafenib treatment efficiency in patients with hepatocellular carcinoma

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World Journal of Gastrointestinal Oncology 2019年第5期11卷 424-435页

作者： Hiroaki Takaya Tadashi Namisaki Naotaka shimozato Kosuke Kaji Mitsuteru Kitade Kei Moriya Shinya Sato Hideto Kawaratani Takemi Akahane Masanori Matsumoto Hitoshi Yoshiji Third Department of Internal Medicine Nara Medical University Department of Blood Transfusion Medicine Nara Medical University

BACKGROUND Many advanced hepatocellular carcinoma(HCC) patients are receiving sorafenib treatment. Sorafenib reportedly improves overall survival(OS) significantly in patients with HCC. Prediction of sorafenib response and prognosis in patients with HCC receiving sorafenib treatment are important due to the potentially serious side effects of sorafenib. A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs 13(ADAMTS13) and von Willebrand factor(VWF) are associated with the pathophysiology of liver cirrhosis and HCC through their roles in hypercoagulability; they are also associated with angiogenesis via vascular endothelial growth factor(VEGF). The imbalance between ADAMTS13 and VWF was associated with prognosis of various cancers in patients undergoing *** To investigate ADAMTS13 and VWF as potential biomarkers for sorafenib response and prognosis in patients with HCC receiving sorafenib *** Forty-one patients with HCC receiving sorafenib treatment were included in this study. The initial daily sorafenib dose was 400 mg in all patients. ADAMTS13 activity(ADAMTS13:AC), VWF antigen(VWF:Ag), VEGF levels were determined by enzyme-linked immunosorbent assay. Univariate andmultivariate analyses were used to determine predictive factors for sorafenib response and prognosis in patients with HCC receiving sorafenib *** ADAMTS13:AC was significantly higher in patients with stable disease(SD),partial response(PR), and complete response(CR) than in those with progressive disease(PD)(P < 0.05). In contrast, VWF:Ag and the VWF:Ag/ADAMTS13:AC ratio were significantly lower in patients with SD, PR, and CR than in those with PD(P < 0.05 for both). Multivariate analysis showed that the VWF:Ag/ADAMTS13:AC ratio was the only predictive factor for sorafenib response and ADAMTS13:AC was the only prognostic factor in patients with HCC receiving sorafenib treatment. The patients with a low ADAMTS13:AC(<78.0) had significantly h

关键词： ADAMTS13 Von Willebrand factor Biomarkers Hepatocellular carcinoma Sorafenib

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Periodic Measurements of Passive Proton Beam Width Using Radiochromic Film in Fixed Gantry System

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International Journal of Medical Physics, Clinical Engineering and Radiation Oncology 2019年第4期8卷 193-203页

作者： Tomohiro shimozato Keisuke Yasui Hireto Kinou Fumiaki Komatsu Faculty of Radiological Technology School of Health Sciences Gifu University of Medical Science Seki Japan Faculty of Radiological Technology School of Health Science Fujita Health University Toyoake Japan Nagoya Proton Therapy Center Nagoya City West Medical Center Nagoya Japan Department of Radiotherapy Gifu University Hospital Gifu Japan

Background and Aim: Irradiation methods such as double scattering method and spot scanning method have been used in proton beam treatment devices. In the scattering method, a ridge filter or a range modulation wheel is used to create a spread-out Bragg peak, but the distribution at the patient position may change due to positional deviation of the incident beam. Therefore, assessment of the incident position of the beam is very important even in the scattering method. To investigate the width and distribution of the proton beam before entering the RMW, a radiochromic film was installed at the outlet of the transport pipe and the entrance of the profile-monitoring detector. Methods: In this study, the distributions of the beam at the exit of the transport pipe and the entrance of the monitor detector were measured using films. The beam width was measured from the full width at half maximum of the profile obtained from the distribution. Measurements were conducted every month for 10 months. Results: Beams of widths ranging from 1.82 to 2.30 mm in the horizontal direction and 4.25 to 5.33 mm in the vertical direction were outputted from the exit of the transport pipe. Beams of widths ranging from 2.16 to 2.67 mm in the horizontal direction and 4.06 to 5.31 mm in the vertical direction were outputted from the entrance of the monitor detector. The maximum width fluctuation for 10 months was 0.55 mm in the horizontal direction and 1.26 mm in the vertical direction at the entrance of the monitor detector. Conclusions: The distribution was obtained before the proton beam was scattered by the scatterer, and then we propose a method to periodically measure and monitor the changes in the beam distributions every month.

关键词： Proton Beam Beam Width Radiochromic Film Periodic Measurement

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Acylcarnitine: Useful biomarker for early diagnosis of hepatocellular carcinoma in non-steatohepatitis patients

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World Journal of Gastrointestinal Oncology 2019年第10期11卷 887-897页

作者： Hiroaki Takaya Tadashi Namisaki Mitsuteru Kitade Naotaka shimozato Kosuke Kaji Yuki Tsuji Keisuke Nakanishi Ryuichi Noguchi Yukihisa Fujinaga Yasuhiko Sawada Soichiro Saikawa Shinya Sato Hideto Kawaratani Kei Moriya Takemi Akahane Hitoshi Yoshiji Third Department of Internal Medicine Nara Medical University

BACKGROUND Early diagnosis of hepatocellular carcinoma(HCC)is necessary to improve the prognosis of ***,the currently available tumor biomarkers are insufficient for the early detection of *** is essential in fatty acid metabolic pathways.A recent study reported that a high level of acylcarnitine may serve as a useful biomarker for the early diagnosis of HCC in steatohepatitis(SH)*** contrast,another study reported that the level of acetylcarnitine(AC2)-one of the acylcarnitine species-in non-SH patients with HCC was decreased vs that reported in those without *** To investigate the usefulness of acylcarnitine as a biomarker for the early diagnosis of HCC in non-SH *** Thirty-three non-SH patients(14 with HCC and 19 without HCC)were enrolled in this *** samples were obtained from patients at the time of *** levels of acylcarnitine and AC2 in the serum were determined through tandem mass *** levels of vascular endothelial growth factor(VEGF)and VEGF receptor 2(VEGFR-2)were determined by enzymelinked immunosorbent *** and multivariate analyses were used to determine early diagnostic factors of *** The level of acylcarnitine was significantly lower in non-SH patients with HCC vs those without HCC(P<0.05).In contrast,the level of lens culinaris agglutininreactive fraction ofα-fetoprotein(AFP)-AFP-L3%-was significantly higher in non-SH patients with HCC vs those without HCC(P<0.05).However,the levels of total carnitine,free carnitine,AFP,des-γ-carboxy prothrombin,VEGF,and VEGFR-2 were not different between patients with and without *** multivariate analysis showed that a low level of acylcarnitine was the only independent factor for the early diagnosis of *** patients with a low level of AC2 had a significantly higher level of VEGF vs those with a high level of AC2(P<0.05).CONCLUSION The metabolic pathways of fatty acids may differ between SH HCC and non-SH HCC

关键词： Acylcarnitine Acetylcarnitine Biomarker Hepatocellular carcinoma Angiogenesis Carnitine palmitoyltransferase 1 Oxidative stress

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Association between ADAMTS13 activity-VWF antigen imbalance and the therapeutic effect of HAIC in patients with hepatocellular carcinoma

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World Journal of Gastroenterology 2020年第45期26卷 7232-7241页

作者： Hiroaki Takaya Tadashi Namisaki Kei Moriya Naotaka shimozato Kosuke Kaji Hiroyuki Ogawa Koji Ishida Yuki Tsuji Daisuke Kaya Hirotestu Takagi Yukihisa Fujinaga Norihisa Nishimura Yasuhiko Sawada Hideto Kawaratani Takemi Akahane Masanori Matsumoto Hitoshi Yoshiji Department of Gastroenterology Nara Medical UniversityKashiharaNara 634-8522Japan Department of Blood Transfusion Medicine Nara Medical UniversityKashiharaNara 634-8522Japan

BACKGROUND Prediction of HAIC treatment response is important for improving the prognosis in patients with hepatocellular carcinoma(HCC).The progression of HCC is related to hypercoagulability and *** is known that ADAMTS13 and von Willebrand factor(VWF)are related to *** addition,previous study reported that the association between ADAMTS13 and VWF,and angiogenesis via vascular endothelial growth factor(VEGF).Recently,ADAMTS13 and VWF have been associated with the prognosis in patients with various kinds of cancer undergoing *** To investigate whether ADAMTS13 and VWF become useful biomarkers of treatment response in HCC patients before the initiation of HAIC *** Seventy-two patients were enrolled in this ***13 activity(ADAMTS13:AC),VWF antigen(VWF:Ag)and VEGF levels were determined via enzyme-linked immunosorbent *** and multivariable analyses were performed to determine the predictive factors of treatment response in patients with HCC undergoing HAIC *** ADAMTS13:AC levels in HCC patients with stable disease(SD)+partial response(PR)of HAIC treatment were significantly higher than those with progressive disease(PD)(P2.7)had higher VEGF levels than those with low ratio(≤2.7).Multivariable analysis revealed that VWF:Ag/ADAMTS13:AC ratio was a predictive factor of HAIC treatment *** VWF:Ag/ADAMTS13:AC ratio may become a useful biomarker of treatment response in HCC patients before the initiation of HAIC treatment.

关键词： ADAMTS13 Von Willebrand factor Vascular endothelial growth factor Biomarkers Hepatocellular carcinoma HAIC

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Measurements of Reactivity Worth of Rare-Earth Elements

Measurements of Reactivity Worth of Rare-Earth Elements

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作者： K.shimozato K.Ieyama T.Kitada H.Okochi A.Yamamoto H.Unesaki Graduate School of Engineering Osaka University Graduate School of Engineering Nagoya University

INTRODUCTION:Rare-earth elements are considered as a candidate of advanced burnable ***, few critical experiments have been carried out so far to validate the accuracy of their nuclear *** critical experiments loaded with rare-earth elements(Dy,Ho,Er and Tm) were carried out and their reactivity worth were measured at the KUCA B *** experiments were performed on B3/8"P36EU(3) core and B1/8"P60EU-EU *** critical experiments attempt to estimate the validity of nuclear data of the rare-earth elements.

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