「選択可能な社会」と社会経済学

departmental bulletin pape

Production of Prompt Charmonia in e+e- Annihilation at sqrt[s] ≈ 10.6 GeV

journal articl

研究速報 : Anisotropic Deformation and Strength Properties of Wet-Tamped Sand in Plane Strain Compression at Low Pressures (Part 2) : Deformation Characteristics at Extremely Small Strain Level

departmental bulletin pape

The Creation of Local Culture by the Local Government in Japan

application/pdfThere are few of example that creation of local culture in small cities in Japan is being developed as a policy, and Nakaniida town is positioned as a pioneer among other cities. The symbol of cultural city construction at Nakaniida town is "Bach Hall". It is a concert hall of specialized classical music built in 1981. In Nakaniida town, there are various devices to produce more influential side effects from one project. As a result, the citizens have sprouted up their affection and pride for the city identity is being established.departmental bulletin pape

A Car Following Model Incorporating Excess Critical Speed Concept

journal articl

Wild-type or catalytic mutant of EcolCdtB and HducCdtB purified under native condition present a similar plasmid digestion activity.

A. SDS-PAGE analysis of WT and D273R (DR) HducCdtB proteins purified from E. coli NiCo21 (DE3) under native conditions. M: molecular weight marker. B. Kinetics of the plasmid digestion assay in presence of WT or D273R HducCdtB. Agarose gel electrophoresis and quantification of supercoiled plasmid (250 ng) incubated with or without 50 ng of WT or D273R HducCdtB for the indicated time. M: molecular weight marker. C. SDS-PAGE analysis of WT and H153A (HA) EcolCdtB proteins purified from E. coli NiCo21 (DE3) under native conditions. M: molecular weight marker. D. Kinetics of the plasmid digestion assay in presence of WT or H153A EcolCdtB. Agarose gel electrophoresis and quantification of supercoiled plasmid (250 ng) incubated with or without 100 ng of WT or H153A EcolCdtB for the indicated times. M: molecular weight marker. E. Ions content effects on plasmid digestion by CdtB. Agarose gel electrophoresis and quantification of supercoiled plasmid (250 ng) incubated in presence or absence of 50 ng of WT or mutant (Hduc D273R or Ecol H153A) CdtB for 2 h, in presence or absence of Mg2+/Ca2+. M: molecular weight marker. B, D and E: arrows indicate plasmid conformation, either relaxed (R), linear (L) or supercoiled (S). For quantifications, the amount of each plasmid conformation is expressed as proportion of the total plasmid content. Results present the mean ± SD of at least three independent experiments; statistical differences were analysed between every conditions and only mutant vs WT comparisons are shown (ns: not significant).</p

DNA damage induction in HeLa cells after CDT holotoxin treatment or mCherry-CdtB expression.

A. Representative images of γH2AX immunofluorescence and DAPI staining from HeLa cells treated with 35 ng/mL of WT or D273R HducCDT holotoxin for 24 h. Scale bar: 50 μm. B. Representative images of γH2AX immunofluorescence and DAPI staining from HeLa cells treated with 2.5 ng/mL of WT or H153A EcolCDT holotoxin for 24 h. Scale bar: 50 μm. C. Quantification of γH2AX signal in HeLa cells left untreated (NT), treated with 20 ng/mL of WT or D273R HducCDT or with 2.5 ng/mL of WT or H153A EcolCDT for 24 h, represented as the mean fluorescence intensity per cell (normalised to 1 for the untreated condition) or as the proportion of γH2AX positive cells. Results present the mean ± SD of at least three independent experiments; statistical differences were analysed between treated and untreated conditions (**** P < 0.0001). D. Representative images of mCherry-HducCdtB localisation, γH2AX immunofluorescence and DAPI staining from HeLa cells expressing WT or D273R HducCdtB in fusion with mCherry. Immunostaining was performed 11 h after transfection. Cells with high expression (stars) or low expression (arrows) of WT mCherry-HducCdtB are indicated. Scale bar: 20 μm. E. Representative images of mCherry-EcolCdtB localisation, γH2AX immunofluorescence and DAPI staining from HeLa cells expressing WT or H153A EcolCdtB in fusion with mCherry. Immunostaining was performed 11 h after transfection. Cells with high expression (stars) or low expression (arrows) of WT mCherry-EcolCdtB are indicated. Scale bar: 20 μm. F. Quantification of γH2AX positive HeLa cells left untransfected (NT), expressing WT or mutant (HducCdtB D273R or EcolCdtB H153A) CdtB in fusion with mCherry. Immunostaining was performed at the indicated time after transfection. Results present the mean ± SD of at least three independent experiments; statistical differences were analysed between transfected and non-transfected conditions at each time point (*** P < 0.001).</p

DNA damage induction in HeLa cells transfected with purified CdtB.

A. Representative images of γH2AX immunofluorescence and DAPI staining from HeLa cells transfected with 50 nM of HducCdtB WT (fractions C2, C11, D4, E5) or D273R (fraction D2) after SEC for 24 h. Scale bar: 20 μm. B. Representative images of γH2AX immunofluorescence and DAPI staining from HeLa cells transfected with 50 nM of WT or D273R HducCdtB before SEC, with 200 nM of DNase I or with 120 nM of HducCdtC for 24 h. Scale bar: 20 μm. C. Representative images of γH2AX immunofluorescence and DAPI staining from HeLa cells transfected with 120 nM of EcolCdtB WT or H153A for 14 h. Scale bar: 20 μm. D. Representative images of γH2AX and 53BP1 immunofluorescence and DAPI staining from HeLa cells transfected with 5 nM of WT or mutant (Hduc D273R or Ecol H153A) CdtB for 14 h. Scale bar: 20 μm. E. Dose-response analysis of γH2AX induction after CdtB transfection. Quantification of γH2AX signal in HeLa cells left untransfected (NT) or transfected with the indicated concentration of WT or mutant (Hduc D273R or Ecol H153A) CdtB or negative controls (HducCdtC or DNase I) for 14 h, represented as the mean fluorescence intensity per cell (normalised to 1 for the untreated condition) or as the proportion of γH2AX positive cells. Results present the mean ± SD of at least three independent experiments. Statistical differences were analysed between transfected and non-transfected conditions for each CdtB concentration (* P < 0.05; **** P < 0.0001) or between HducCdtB and EcolCdtB (# P < 0.05; ## P < 0.01; #### P < 0.0001). F. Kinetics of γH2AX induction after CdtB transfection. Quantification of γH2AX signal in HeLa cells left untransfected (NT) or transfected with 120 nM of WT EcolCdtB or HducCdtB for the indicated time, represented as the mean fluorescence intensity per cell (normalised to 1 for the untreated condition) or as the proportion of γH2AX positive cells. Results present the mean ± SD of at least three independent experiments; statistical differences were analysed between transfected and non-transfected conditions for each time point (* P < 0.05; ** P < 0.01; ***P < 0.001; **** P < 0.0001) or between HducCdtB and EcolCdtB (# P < 0.05; ## P < 0.01; ### P < 0.001; #### P < 0.0001).</p

Wild-type or mutant HducCdtB proteins purified under denaturing conditions present a similar plasmid digestion activity.

A. SDS-PAGE analysis of WT and D273R (DR) HducCdtB proteins purified from E. coli NiCo21 (DE3) under denaturing conditions. B. Kinetics and CdtB concentration effect on plasmid digestion assay with WT or D273R HducCdtB. Agarose gel electrophoresis and quantification of supercoiled plasmid (250 ng) incubated in presence of the indicated concentration of WT or D273R HducCdtB or bovine DNase I for the indicated times. C. Size exclusion chromatography (SEC) of WT HducCdtB. The curve represents the absorbance at 280 nm of the fractions eluted from the SEC column. The fractions used for further analysis are shown with black arrows. The selected fractions were analysed by Western Blot with anti-HIS antibody. The peak fractions from WT and D273R (DR) HducCdtB were analysed by Western Blot with anti-HIS antibody. D. Plasmid digestion assay with WT or D273R HducCdtB purified from SEC. Agarose gel electrophoresis and quantification of supercoiled plasmid (125 ng) incubated in presence of 1 μg of WT (D4) or D273R (D2) HducCdtB or in control conditions (no protein, with 2 ng of bovine DNase I or with 1 μg of CdtC) for 7 h in presence or in absence of Mg2+/Ca2+ buffer. B and D: arrows indicate plasmid conformation, either relaxed (R), linear (L) or supercoiled (S). For quantifications, the amount of each plasmid conformation is expressed as proportion of the total plasmid content. Results present the mean ± SD of at least three independent experiments, except for the D4 and D2 fractions without ions which were replicated twice; statistical differences were analysed between every conditions and only mutant vs WT comparisons are shown (ns: not significant).</p

Food-grade titanium dioxide translocates across the buccal mucosa in pigs and induces genotoxicity in an <i>in vitro</i> model of human oral epithelium

The whitening and opacifying agent titanium dioxide (TiO2) is used worldwide in various foodstuffs, toothpastes and pharmaceutical tablets. Its use as a food additive (E171 in EU) has raised concerns for human health. Although the buccal mucosa is the first area exposed, oral transmucosal passage of TiO2 particles has not been documented. Here we analyzed E171 particle translocation in vivo through the pig buccal mucosa and in vitro on human buccal TR146 cells, and the effects on proliferating and differentiated TR146 cells. In the buccal floor of pigs, isolated TiO2 particles and small aggregates were observed 30 min after sublingual deposition, and were recovered in the submandibular lymph nodes at 4 h. In TR146 cells, kinetic analyses showed high absorption capacities of TiO2 particles. The cytotoxicity, genotoxicity and oxidative stress were investigated in TR146 cells exposed to E171 in comparison with two TiO2 size standards of 115 and 21 nm in diameter. All TiO2 samples were reported cytotoxic in proliferating cells but not following differentiation. Genotoxicity and slight oxidative stress were reported for the E171 and 115 nm TiO2 particles. These data highlight the buccal mucosa as an absorption route for the systemic passage of food-grade TiO2 particles. The greater toxicity on proliferating cells suggest potential impairement of oral epithelium renewal. In conclusion, this study emphasizes that buccal exposure should be considered during toxicokinetic studies and for risk assessment of TiO2 in human when used as food additive, including in toothpastes and pharmaceutical formulations.</p