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| 1.
学位論文 |
学位
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Oishi, Takuma ; Sasaki, Aatsushi ; Hamada, Nobuyuki ; Ishiuchi, Shogo ; Funayama, Tomoo ; Sakashita, Tetsuya ; Kobayashi, Yasuhiko ; Nakano, Takashi ; Nakazato, Yoichi
概要:
application/pdf<br />Thesis or Dissertation<br />Histological analyses of glioblastoma cells after carbon-ion exposure a
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re still limited and ultrastructural characteristics have not been investigated in detail. Here we report the results of morphological and morphometric analyses of a human glioblastoma cell line, CGNH-89, after ionizing radiation to characterize the effect of a carbon-beam on glioblastoma cells. Using CGNH-89 cells exposed to 0–10 Gy of X-ray (140kVp) or carbon-ions (18.3 MeV/nucleon, LET = 108 keV/μm), we performed conventional histology and immunocytochemistry with MIB-1 antibody, transmission electron microscopy, and computer-assisted, nuclear size measurements. CGNH-89 cells with a G to A transition in codon 280 in exon 8 of the TP53 gene had nuclei with pleomorphism, marked nuclear atypia and brisk mitotic activity. After carbon-ion and X-ray exposure, living cells showed decreased cell number, nuclear condensation, increased atypical mitotic figures, and a tendency of cytoplasmic enlargement at the level of light microscopy. The deviation of the nuclear area size increased during 48 hours after irradiation, while the small cell fraction increased in 336 hours. In glioblastoma cells of the control, 5 Gy carbon-beam, and 10 Gy carbon-beam, and MIB-1 labeling index decreased in 24 hours (12%, 11%, 7%, respectively) but increased in 48 hours (10%, 20%, 21%, respectively). Ultrastructurally, cellular enlargement seemed to depend on vacuolation, swelling of mitochondria, and increase of cellular organelles, such as the cytoskeleton and secondary lysosome. We could not observe apoptotic bodies in the CGNH-89 cells under any conditions. We conclude that carbon-ion irradiation induced cell death and senescence in a glioblastoma cell line with mutant TP53. Our results indicated that the increase of large cells with enlarged and bizarre nuclei, swollen mitochondria, and secondary lysosome occurred in glioblastoma cells after carbon-beam exposure.<br />学位記番号：医博甲1096
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| 2.
論文 |
論文
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Ikota, Hayato ; Kinjo, Sawako ; Yokoo, Hideaki ; Nakazato, Yoichi
概要:
application/pdf<br />Journal Article<br />We performed a systematic immunohistochemical study on 378 brain tumors using 37 antibodies and tissue microarray (TMA) technology. The aim of this study was to find new diagnostic
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biomarkers using antibodies established in our laboratory. Our TMA consisted of a grid of 1.5-mm cores that were extracted from individual donor blocks. Staining for each antibody was scored using a three-point system. We used hierarchical clustering analysis to interpret these data, which resulted in separation of all the brain tumors into seven groups. Although there were some exceptions, cases with the same histological diagnosis were generally grouped together. We then carried out statistical analyses to find the most useful antibodies for grouping of brain tumors. Ten antibodies [glial fibrillary acidic protein (GFAP), Olig2, vimentin, epithelial membrane antigen (EMA), cytokeratin (AE1/AE3), alpha-internexin, nestin, pinealocytes PP5, aquaporin-4 (AQP4) M13d and AQP4M13e] discriminated between astrocytomas and oligodendroglial tumors. Six antibodies [EMA, AE1/AE3, TUJ1, nestin, neurofilament protein-MH (NF-MH) and perivascular cells GP-1] showed significant differences between high-grade and low-grade gliomas. Our data have revealed new antibodies with potential diagnostic utility (Olig2, PP5, GP-1) and demonstrate that TMA technology is highly useful for evaluating newly established antibodies in brain-tumor research.<br />The original publication is available at www.springerlink.com
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| 3.
論文 |
論文
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Ito, Yoshikazu Z. ; Hiromachi, Sachiko ; Futawatari, Tamae ; Arai, Michiko ; Nakazato, Yoichi
概要:
application/pdf<br />Departmental Bulletin Paper<br />A tumorigenic epithelial-like cell line of rat prostate origin (PR4) was established in vitro. The cell line originated from a single cell of a previously established rat
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ventral prostate cell line, and has been serially subcultured more than 100 times over 5 years. The doubling time of the cells was 18 hours, and the saturation density was 5 x 10(6) cells/ml. Oval nuclei with dispersed chromatin and well developed cytoplasmic organelles were ultrastructural characteristics of the cells. The PR4 cells were transplanted into nude mice and the histology of the resulting tumors was neopolastic epithelial-like morphology that showed cells arranged in a sheet and cord structure with a solid pattern. Very high activities of prostatic acid phosphatase and beta-glucuronidase were observed in the cytosol of the cells. The PR4 cells did not respond to androgen. The use of the new cell line and other in vitro models of rat prostate as experimental tools for human prostatic cancer research was discussed.
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| 4.
論文 |
論文
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Yamaguchi, Haruyasu ; Haga, Chie ; Hirai, Shunsaku ; Nakazato, Yoichi ; Kosaka, Kenji
概要:
application/pdf<br />Departmental Bulletin Paper<br />We developed a new type of methenamine silver (MS) stain for senil
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e plaques (SP) on paraffin-embedded tissue sections. This method is a modification of both Gomori's methenamine silver nitrate, and Jone's periodic acid-methenamine silver (PAM) stains. We omitted chemical preoxidation by either periodic acid in the PAM stain or chromic acid in Gomori's stain, and determined the best condition of silver solution for SP. This resulted in selective staining of SP and amyloid angiopathy, because capillary basement membranes, corpora amylacea, macrophage granules, and nerve cell pigments remained unstained. The argyrophilia of SP was not affected by pretreatment with aldehyde blocking reagents. In comparison with βprotein immunostain, our MS stain showed almost the same staining pattern as that of the βprotein immunostain. Moreover, the argyrophilia of SP was selectively abolished by formic acid pretreatment, which has been shown to destroy β-pleated sheet structure of amyloid, suggesting that our MS stain impregnates amyloid-related components of SP. Our MS stain is a new, rapid, easy, and reliable method, and sensitive enough for routine or screening studies of the SP.
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| 5.
論文 |
論文
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Yamaguchi, Haruyasu ; Hirai, Shunsaku ; Nakazato, Yoichi
概要:
application/pdf<br />Departmental Bulletin Paper<br />We studied the effect of formic acid pretreatment on silver stains
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and βprotein immunostain on brain tissue samples from patients with the Alzheimer-type dementia (AID). The argyrophilia of the amyloid component of senile plaques (SP), which could be visualized by modified Bielschowsky and periodic acid-methenamine silver (PAM) stains, was not visible when the specimens were pretreated with formic acid. In contrast, the pretreatment highly enhanced the immunoreactivity of this component with βprotein antiserum. Our findings suggest that formic acid pretreatment selectively abolishes the argyrophilia of the amyloid component of SP.
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