Original Article

本研究の目的は,健康な小児ががんや白血病といった病名からどのようなイメージを抱くのか,自分が大きな病気になったとしたら,病名や治療などについて教えて欲しいと思っているのかを明らかにすることである。千葉県内の小中高校に通っている小学校5年生から高校3年生までの児童生徒1964名から得られたアンケートの回答を,統計ソフトSPSSにて分析を行い以下の結果を得た。1.健康な小児が抱くがんのイメージは,「死ぬ・治らない」「重い・危険・苦しい」といった悲観的なのものが多く,全体の約65%を占めていた。「聞いたことがない・わからない」と答えたものもおり全体の約15%であり,どの学年でも同様の傾向であった。2.健康な小児が抱く白血病のイメージは「病態や特徴について」「原因や治療について」といったものが,全体の約38%を占めていた。「聞いたことがない・わからない」と答えたものは全体の約33%を占め,小学生では半数以上が「聞いたことがない・わからない」と答えていた。3.自分が病気になったときにされる説明については小学生の80%,中学生の85%,高校生の91%が真実を伝えられることを求めていた。その理由として自分の知る権利,治療に前向きに取り組めるといった姿勢の向上,知らないことが不安になる,残された命を有意義に悔いのないように過ごしたいといったものがあげられていた。The purposes of this study were to identify the images of healthy children on cancer and leukemia, and the way of thinking of truth telling with disease. The number of subjects were 1964. They belonged to between the fifth grade of primary school and the third year in high school. They answered someitem-questionnaire, and the answers were analyzed using SPSS. The results were as follows: 1. Sixty-five percent of the images of cancer were pessimistic, like death or incurable and serious or painful . About fifteen percent were having no images. 2. Thirty-eight percent of the images of leukemia were the feature and the cause or the treatment . About thirty-three percent were having no images. 3. Truth telling was desired by eighty percent of students of primary schools, eighty-five percent of junior high schools and ninety-one percent of a high school. The reasons were the rights to know , to be patient with treatments , to become more anxious without truth telling , and to live the remaining days without regrets

Acetolactate synthase regulatory subunits play divergent and overlapping roles in branched-chain amino acid synthesis and Arabidopsis development

Branched-chain amino acids (BCAAs) are synthesized by plants, fungi, bacteria, and archaea with plants being the major source of these amino acids in animal diets. Acetolactate synthase (ALS) is the first enzyme in the BCAA synthesis pathway. Although the functional contribution of ALS to BCAA biosynthesis has been extensively characterized, a comprehensive understanding of the regulation of this pathway at the molecular level is still lacking

The SKP1-Like Gene Family of Arabidopsis Exhibits a High Degree of Differential Gene Expression and Gene Product Interaction during Development

The Arabidopsis thaliana genome encodes several families of polypeptides that are known or predicted to participate in the formation of the SCF-class of E3-ubiquitin ligase complexes. One such gene family encodes the Skp1-like class of polypeptide subunits, where 21 genes have been identified and are known to be expressed in Arabidopsis. Phylogenetic analysis based on deduced polypeptide sequence organizes the family of ASK proteins into 7 clades. The complexity of the ASK gene family, together with the close structural similarity among its members raises the prospect of significant functional redundancy among select paralogs. We have assessed the potential for functional redundancy within the ASK gene family by analyzing an expanded set of criteria that define redundancy with higher resolution. The criteria used include quantitative expression of locus-specific transcripts using qRT-PCR, assessment of the sub-cellular localization of individual ASK:YFP auto-fluorescent fusion proteins expressed in vivo as well as the in planta assessment of individual ASK-F-Box protein interactions using bimolecular fluorescent complementation techniques in combination with confocal imagery in live cells. The results indicate significant functional divergence of steady state transcript abundance and protein-protein interaction specificity involving ASK proteins in a pattern that is poorly predicted by sequence-based phylogeny. The information emerging from this and related studies will prove important for defining the functional intersection of expression, localization and gene product interaction that better predicts the formation of discrete SCF complexes, as a prelude to investigating their molecular mode of action

Relationship of the <i>ASK</i> gene family in Arabidopsis.

<p>The phylogenetic grouping of <i>ASK</i> genes based on their deduced primary amino acid sequence was calculated using the NJ method described. The genes are grouped into seven distinct clades as denoted by the vertical lines. Numbers at the branches represents percentage bootstrap support calculated for a 1000 replicates. All tree branches are scaled to the number of amino acid substations per site.</p

Relative organ-specific abundance of cDNAs for select ASK genes.

<p>All gene expression was normalized relative to <i>ACTIN2</i> expression. <b>A</b>; Relative abundance of <i>ASK</i> gene cDNAs in flowers from stage 9 plants relative to seedlings. <b>B</b>; Relative abundance of <i>ASK</i> cDNAs in stage 15 flowers relative to seedlings. <b>C</b>; Relative expression of <i>ASK</i> genes in siliques relative to seedlings.</p

Interaction profile of select ASK and F-Box proteins as assessed using BiFC.

<p>Visualization of BiFC-based sub-cellular protein interactions between select ASK and F-Box proteins (TIR1 or UFO) in <i>N. benthamiana</i> epidermal leaf cells: YFP signal indicates a positive interaction; chlorophyll auto-fluorescence is shown in red. <b>A–H</b>; protein interactions between TIR1 and the ASK1,3,4,9 proteins expressed as BiFC fusion expression constructs. <b>I–P</b>; protein interactions between UFO and ASK1,3,4,9 proteins expressed as BiFC fusion expression constructs. <b>Q;</b> The indicated interaction map was developed as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0050984#s2" target="_blank">methods</a>, and summarizes the results obtained following transient expression of BiFC constructs in <i>N. benthamiana</i> leaf epidermal cells. Edge lines joining nodes represent a positive interaction.</p

Confocal imaging and sub-cellular localization of ASK proteins in transgenic Arabidopsis. A&C

<p>; Propidium iodide-stained epidermal root cell walls or leaf epidermal cell walls and guard cell nuclei. <b>B</b>; Chlorophyll auto-fluorescence from the leaf mesophyll cell layer. <b>D–F</b>; stable expression of N-terminal YFP-tagged ASK3 protein in transgenic Arabidopsis. <b>G–I</b>; merged channels corresponding to panels (<b>A–C</b>) and (<b>D–F</b>).</p

Evaluation of oligomerization potential of SCF^TIR1 subunits.

<p><b>(a-d)</b> BiFC-based evaluation of TIR1, CUL1, ASK1 oligomerization and ASK1-TIR1 interaction, respectively, following transient expression in Nicotiana leaves. <b>(a'-d')</b> Propidium iodide staining of the nucleus. <b>(e)</b> Assessment of TIR1-TIR1 protein interaction in a Y2H assay. Images of single colonies expressing the designated constructs and grown on histidine plates (top panel) and test plates containing the indicated concentrations of 3-AT in the absence of histidine (bottom panels). <b>(f)</b> Co-IP-based assessment of TIR1 oligomerization following transient expression in Nicotiana leaves. HA:TIR1 and Myc:TIR1 were co-injected in leaves. Protein extracts were subjected to immunoprecipitation using anti-Myc antibody. Immunoprecipitates were examined by western-blotting using anti-Myc and anti-HA antibodies. (<b>g</b>) Co-IP-based assessment of TIR1 oligomerization in double <i>pTIR1</i>:<i>TIR1–GUS</i> and <i>pTIR1</i>:<i>gTIR1–VENUS</i> transgenic Arabidopsis plants. Protein extracts were subjected to immunoprecipitation using anti-GUS antibody. Immunoprecipitates were examined by western-blotting using anti-GUS and anti-GFP antibodies.</p

A set of spatially-clustered amino acids are critical for TIR1 oligomerization.

<p><b>(a, c, e, g, i</b> and <b>k)</b> BiFC-based assessment of TIR1 oligomerization using wild-type and G142A, F143A, I151A, V117A and G147A mutants, respectively, following transient expression in Nicotiana leaves. <b>(b, d, f, h, j</b> and <b>l)</b> Assessment of TIR1-ASK1 interaction using wild-type and G142A, F143A, I151A, V117A and G147A mutants respectively, following transient expression in Nicotiana leaves. <b>(m</b> and <b>n)</b> Assessment of TIR1 interaction with D170A and D170E mutants using BiFC. <b>(o)</b> Co-IP experiments in Nicotiana leaves. HA-tagged wild-type and mutant Flag tagged TIR1 constructs were co-injected. Protein extracts were subjected to immuno-precipitation using anti-HA conjugated beads. The immune-precipitates were examined by western-blotting using anti-Flag and anti-HA antibodies. <b>(p)</b> Side view of TIR1-IAA7 peptide structure (PDB 2P1Q); residues critical for TIR1 oligomerization are depicted in orange.</p