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Articles by S Yamashita
Total Records ( 3 ) for S Yamashita
  H Takeshima , S Yamashita , T Shimazu , T Niwa and T. Ushijima

Instructive mechanisms are present for induction of DNA methylation, as shown by methylation of specific CpG islands (CGIs) by specific inducers and in specific cancers. However, instructive factors involved are poorly understood, except for involvement of low transcription and trimethylation of histone H3 lysine 27 (H3K27me3). Here, we used methylated DNA immunoprecipitation (MeDIP) combined with a CGI oligonucleotide microarray analysis, and identified 5510 and 521 genes with promoter CGIs resistant and susceptible, respectively, to DNA methylation in prostate cancer cell lines. Expression analysis revealed that the susceptible genes had low transcription in a normal prostatic epithelial cell line. Chromatin immunoprecipitation with microarray hybridization (CHiP-chip) analysis of RNA polymerase II (Pol II) and histone modifications showed that, even among the genes with low transcription, the presence of Pol II was associated with marked resistance to DNA methylation (OR = 0.22; 95% CI = 0.12–0.38), and H3K27me3 was associated with increased susceptibility (OR = 11.20; 95% CI = 7.14–17.55). The same was true in normal human mammary epithelial cells for 5430 and 733 genes resistant and susceptible, respectively, to DNA methylation in breast cancer cell lines. These results showed that the presence of Pol II, active or stalled, and H3K27me3 can predict the epigenetic fate of promoter CGIs independently of transcription levels.

  S Yamashita , O Katsumata and Y. Okada

We have developed a new standardized method for the post-embedding immunoelectron microscopy using the same fixation, antigen retrieval and image contrasting procedures. Tissues were fixed with 4% formaldehyde containing 2.5 mM CaCl2, 1.25 mM MgCl2 in a 0.1 M 4-(2-hydroxyethyl)-piperazineethanesulfonic acid (HEPES) buffer (pH 7.4) for 2 h and then with the same fixative composition in 0.1 M HEPES buffer (pH 8.5) overnight at room temperature. Vehicle osmolarity of fixatives was adjusted to 300–330 mOsm by adding glucose. The specimens were dehydrated with dimethylformamide on ice and embedded in LR-White resin. Ultrathin sections were heated in a 20 mM Tris–HCl buffer (pH 9.0) for 1–2 h at 95°C. After immuno-gold labeling, the sections were treated with 2% glutaraldehyde containing 0.05% tannic acid in a 0.1 M phosphate buffer (pH 5.5) for 5 min and with a 1% OsO4/0.1 M phosphate buffer (pH 7.4) for 5 min, and then they were double stained with uranyl acetate and lead citrate. The standardized method yielded strong and reproducible immunoreactions for soluble, membrane-bound and filamentous proteins showing an excellent image contrast without destruction of the fine structures.

  Y Sekitani , N Hayashida , I. V Karevskaya , O. A Vasilitsova , A Kozlovsky , M Omiya , S Yamashita and N. Takamura

To determine the current risk of internal radiation exposure after the Chernobyl accident, the 137Cs body burden of the inhabitants of Bryansk Oblast, Russian Federation was evaluated, from 1998 to 2008. The study population is composed of 84 666 people who visited Bryansk No. 2 Hospital. A whole-body counter was used for measurement of 137Cs body burden. 137Cs concentration was significantly higher in the late period during the study and showed seasonal variation, suggesting that inhabitants may have consumed contaminated forest products. However, people with high annual exposure doses were quite rare during all years of the study. In conclusion, although internal radiation exposure from 137Cs continues to this day in Bryansk Oblast, the annual exposure dose is low in almost all inhabitants. Because of the long half-life of 137Cs, the long-term follow-up is necessary to monitor the health status and relieve the anxiety of the inhabitants around Chernobyl.

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