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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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