Observation of b→dγ and Determination of |Vtd/Vts|

journal articl

Antagonistic effects of ethyl methanesulfonate and maleic hydrazide in inducing somatic mutations in the stamen hairs of Tradescantia clone BNL 4430

Mutagenic interaction between ethyl methanesulfonate (EMS; a monofunctional alkylating agent) and maleic hydrazide (MH; a promutagen activated into a mutagen in plants highly likely by peroxidase) was studied in the stamen hairs of Tradescantia clone BNL 4430, a blue/pink heterozygote. Since EMS has been shown to act synergistically with X rays in inducing mutations, and mutagenic synergisms have also been observed between X rays and MH by exposing to X rays before MH treatments, EMS and MH were expected to act synergistically at least by exposing to EMS before MH treatment. Three different combined treatments were conducted, i.e., by exposing for 4 h to 18.8 mM EMS 44 or 20 h before starting or 20 h after completing 1 mM MK treatments for 4 h. Unexpectedly, however, clear antagonistic effects in inducing somatic pink mutations were detected after all these combined treatments. Especially, the induced mutation frequency by exposing to EMS 44 h before the MH treatment was significantly lower than that induced by MH alone. The clear mutagenic antagonisms observed were thought to have resulted from EMS-caused inhibition of activation of MH by peroxidase, EMS ethylating and thus inactivating this enzyme or its precursors. Decreased peroxidase activities than those after treatments with MH alone were measured after two combined treatments, i.e., 12 h after the one exposing to EMS 44 h before MH and 24 h after the other exposing to EMS 20 h after MH, but the decreases were not large enough or their fluctuations were too large to judge them to be statistically significant. Comparisons of mutation frequencies induced by the combined treatments exposing to EMS before MH with those by MH alone suggest that there are some mechanisms (other than ethylation of peroxidase or its precursors) by which EMS suppresses the activation of MH.textapplication/pdfjournal articl

労働紛争の解決システムに関する日独法比較（解題）

2008-03-25departmental bulletin pape

大城炭坑山計画

東京帝国大学工科大学種別:卒業論文thesi

Regulation of inflorescence architecture by intertissue layer ligand-receptor communication between endodermis and phloem

Multicellular organisms achieve final body shape and size by coordinating cell proliferation, expansion, and differentiation. Loss of function in the Arabidopsis ERECTA (ER) receptor-kinase gene confers characteristic compact inflorescence architecture, but its underlying signaling pathways remain unknown. Here we report that the expression of ER in the phloem is sufficient to rescue compact er inflorescences. We further identified two EPIDERMAL PATTERNING FACTOR-LIKE (EPFL) secreted peptide genes, EPFL4 and EPFL6/CHALLAH (CHAL), as redundant, upstream components of ER-mediated inflorescence growth. The expression of EPFL4 or EPFL6 in the endodermis, a layer adjacent to phloem, is sufficient to rescue the er-like inflorescence of epfl4 epfl6 plants. EPFL4 and EPFL6 physically associate with ER in planta. Finally, transcriptome analysis of er and epfl4 epfl6 revealed a potential downstream component as well as a role for plant hormones in EPFL4/6- and ER-mediated inflorescence growth. Our results suggest that intercell layer communication between the endodermis and phloem mediated by peptide ligands and a receptor kinase coordinates proper inflorescence architecture in Arabidopsis.journal articl

Pharmacokinetics of NKp30-Ig and NKp46D2-Ig fusion proteins <i>in vivo.</i>

<p>Mice were injected i.p. with one dose (5mg/kg) of NKp30-Ig (a) or NKp46D2-Ig (b). Serum sample were collected (at 0, 0.5, 1, 2, 6, 25, 48, 120, 168, 264 and 312 hours after injection) and levels of NKp30-Ig or NKp46D2-Ig were determined in a standard ELISA assay. Figure shows the average amount of fusion proteins detected in the serum of three mice, measure at each time point. Error bars represent mean±s.d of triplicates.</p

Mechanism of NKp30-Ig mediated tumor regression.

<p>(a,b) NKp30-Ig does not induce apoptosis in tumor cells. PC3/<i>luc</i> (a) or DU145 (b) cells were incubated with increasing concentrations of NKp30-Ig, NKp46D2-Ig, control Ig or PBS in the presence of a cross-linking antibody. After 48 hours, the percentage of apoptotic cells was determined by Annexin V and PI staining. The figure shows one out of three experiments performed. (c,d) NKp30-Ig can mediate tumor opsonization by macrophages. Radioactive labeled PC3/<i>Luc</i> (c) or DU145 (d) cells were incubated with LPS-activated macrophages at the indicated E∶T ratios. Specific lysis was determined after 48 hours. Error bars represent mean±s.d of triplicates. Figure represents one out of three experiments performed. (e) Infiltration of macrophages to the tumor tissue. Human prostate tumors (DU145) grown in nude mice were fixed in 10% buffered formalin. Paraffin-embedded sections were stained in Hematoxylin and Eosin. The arrows indicate tumor associated- macrophages (X380). This figure represents one out of 5 sections tested.</p

Expression of NCRs ligands on human prostate cancer.

<p>(a,b) NKp30-Ig and NKp46D2-Ig specifically bind to human prostate cell lines. PC3/<i>Luc</i> (a) and DU145 (b) cell lines were stained with NKp30-Ig, NKp46D2-Ig or control CD99-Ig, followed by PE-conjugated mouse anti-human IgG1 antibody. Grey histograms represent the background staining by the control CD99-Ig fusion protein and the black empty histograms represent the staining by either NKp30-Ig or NKp46D2-Ig, as indicated in the top of each histogram. This figure represents one experiment out of three performed. (c) Immunohistochemical staining of primary human prostate adenocarcinoma and benign prostate hyperplasia (BPH) by NKp30-Ig and NKp46D2-Ig. Cuts from formalin-fixed and paraffin-embedded human prostate adenocarcinoma (upper panel) and BPH (lower panel) were antigen-retrieved by microwave-citrate treatment. Slides were then stained with NKp30-Ig, negative control CD99-Ig or NKp46D2-Ig, followed by biotinylated-goat-anti-human-Fc and avidin-biotin HRP complex. Substrate for HRP was AEC (red color) and slides were counter-stained with Hematoxylin. Figure shows a representative staining at X400 magnification. Arrow in NKp30-Ig staining of adenocarcinoma (top left panel) points to a representative membrane staining. Staining intensity for top left and top right panels is considered as 2 (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002150#s2" target="_blank">Methods</a>). (d) Expression of NKp30 and NKp46 ligands is abundant on malignant prostate tumors. Cuts from different patients suffering from benign (<i>n</i> = 8) or malignant (<i>n</i> = 9) prostate tumors were prepared and stained as above. Staining was performed in triplicates. Analysis of staining intensity (0-3) and percentage of stained tumor cells was performed by two pathologists. Positive staining was defined when staining intensity was above 1 and encompassed at least 50% of the cells, as described in ‘<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002150#s2" target="_blank">material and methods</a>’.</p

Summary of fusion protein treatment.

<p>(a) Visualization of tumor progression and distribution <i>in vivo</i>. The figure shows an image visualization of one representative animal of each treatment. The scale on the right of each figure describes the color map of the photon count. The integrated light emission (‘I’) is indicated in the left of each photo. (b) Summary of treatment effect. Table describes the overall effect of treatments, as shown in details in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002150#pone-0002150-g003" target="_blank">figure 3</a>.</p

Treatment with NKp30-Ig reduces PC3/<i>Luc</i> tumor growth.

<p>Male nude mice were injected with PC3/<i>luc</i> tumor cells (15×10⁶) into the SC left flanks. Three weeks after tumor implantation, mice were injected (i.p.) every second day over a period of one month with 4mg/kg of NKp30-Ig (<i>n</i> = 16) (a), NKp46D2-Ig (<i>n</i> = 9) (b) or PBS (<i>n</i> = 8) (c). Tumor progression was monitored by measuring light emission from each individual mouse in the initiation (‘start point’) and in the end (‘end point’) of the treatment period. Y-axes represent the relative (in percentage) changes in tumor size after treatment, as calculated from the integrated light emission measured in each time point (indicated numbers above columns). Shrinkage of the tumor by 20% or below its original size was referred as ‘efficient treatment’. This figure is a summary of two experiments and includes all the mice that were tested.</p