High-affinity uptake of kynurenine and nitric oxide-mediated inhibition of indoleamine 2,3-dioxygenase in bone marrow-derived myeloid dendritic cells

Indoleamine 2,3-dioxygenase (IDO)-initiated tryptophan metabolism along the kynurenine (Kyn) pathway in some dendritic cells (DC) such as plasmacytoid DC regulates T-cell responses. It is unclear whether bone marrow-derived myeloid DC (BMDC) express functional IDO. The IDO expression was examined in CD11c+CD11b+ BMDC differentiated from mouse bone marrow cells using GM-CSF. CpG oligodeoxynucleotides (CpG) induced the expression of IDO protein with the production of nitric oxide (NO) in BMDC in cultures for 24 hr. In the enzyme assay using cellular extracts of BMDC, the IDO activity of BMDC stimulated with CpG was enhanced by the addition of a NO synthase (NOS) inhibitor, suggesting that IDO activity was suppressed by NO production. On the other hand, the concentration of Kyn in the culture supernatant of BMDC was not increased by stimulation with CpG. Exogenously added Kyn was taken up by BMDC independently of CpG stimulation and NO production, and the uptake of Kyn was inhibited by a transport system L-specific inhibitor or high concentrations of tryptophan. The uptake of tryptophan by BMDC was markedly lower than that of Kyn. In conclusion, IDO activity in BMDC is down-regulated by NO production, whereas BMDC strongly take up exogenous Kyn.journal articl

アジアの日本語幼稚園に通う幼児の異文化体験と異文化適応<教育科学>

Recently, the number of overseas resident businesspeople from Japan in Asian countries has been increasing. This study aims to examine methods to support childcare in families of such businesspeople and collect basic data for planning the content of education at Japanese kindergartens. A questionnaire survey was conducted with parents rearing young children (N = 360, 211 boys and 149 girls, mean age, 4 years and 10 months) attending Japanese kindergartens in Bangkok and Shanghai. The questionnaire included the following items: children’s experiences of different cultures, their feelings toward different cultures, and their cultural adaptation from the perspective of parents, as well as the types of parents’ involvement when children experienced cultural differences. The results indicated the following. Regardless of age, children did not have deep intercultural contacts, whereas they had shallow intercultural experiences such as having a meal at restaurants where local people gathered. On the other hand, they had many contacts with places and things related to Japanese people and Japanese language. These tendencies became weaker as the length of stay became longer. Furthermore, children felt cultural differences in languages, meals, life habits, and atmosphere of the cities. Such feelings were negative in general, though negative feelings became weaker as the length of stay became longer, except in the case of five-year-old children. Children’s positive orientation toward different cultures, which shows adaptation to different cultures, became significantly stronger as they have more intercultural experiences and their positive feelings toward different cultures became stronger. In five-year-old children, the length of stay was related to low adaptation to different cultures. The above results suggest that it would be important to provide educational support for children so that they could have positive intercultural contacts at Japanese kindergartens and in their homes, and they could recognize experiences of cultural differences positively. Furthermore, difficulties in education of five-year-old children in foreign cultures were also suggested. It would be important to establish cooperative systems between Japanese kindergartens and Japanese elementary schools, and between Japanese kindergartens and local kindergartens as well as local elementary schools.textapplication/pdfdepartmental bulletin pape

Upper Bound on the Decay τ→μγ from the Belle Detector

journal articl

Exams test design and number of selected students by course, master/scoring method, examination method and gender.

Exams test design and number of selected students by course, master/scoring method, examination method and gender.</p

Results of multiple linear regression for predicting exam score.

Results of multiple linear regression for predicting exam score.</p

Immunolocalization of overexpressed hClC-6 in SH-SY5Y cells.

<p>Double immunofluorescence confocal images of SH-SY5Y cells, transiently transfected with pcDNA3.1(−)/hClC-6a expression vector. Overexpression levels were very high, so that transfected cells could easily be distinguished from non-transfected cells. Overexpressed hClC-6 (left column) was detected using the polyclonal α-hClC-6 antibody and visualized with anti-rabbit IgG antibodies conjugated to Alexa Fluor 488 (green signal). Markers for different endosomal compartments (middle column) were (A) mouse anti-EEA-1 (an early endosome marker); (B) mouse anti-transferrin receptor (TfR, an early/recycling endosome marker); (C) mouse anti-LAMP-1 (a late endosomal/lysosomal marker). Primary antibodies were visualized using anti-mouse IgG antibodies conjugated to Alexa Fluor 594 (red signal). In the merged pictures (right column) colocalization is indicated by a yellow signal. The scale bars represent 10 µm.</p

Percentage of students that show different answering patterns on at least one multiple-choice question.

Percentage of students that show different answering patterns on at least one multiple-choice question.</p

Student responses on stress-related questions on elimination testing with adapted scoring and negative marking after receiving the exam score.

Student responses on stress-related questions on elimination testing with adapted scoring and negative marking after receiving the exam score.</p

hClC-6 is glycosylated upon overexpression.

<p>(A) Western blots showing the effect of tunicamycin and (a) PNGaseF and (b) EndoH on hClC-6. For tunicamycin COS-1 cells were incubated with tunicamycin (0.05 and 0.1 µg/ml) for 36 hours; for PNGaseF and EndoH membrane fractions of hClC-6 expressing COS-1 cells were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000474#s2" target="_blank">materials and methods</a>. ‘WT’ refers to untreated hClC-6. The small difference in molecular mass between PNGaseF and tunicamycin-treated hClC-6 might be due to the presence of oligosaccharides carrying fucose-linked α1–3 to the GlcNac attached directly to asparagines, which are PNGaseF resistant as described by Dwek <i>et al. </i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000474#pone.0000474-Dwek1" target="_blank">[53]</a>. (B)(a), Model of the hClC multimeric structure of 2 homologous subunits with the possible glycosylation sites marked as spheres. (b), Partial sequence alignment (ClustalW) of all members of the CLC family reveals that Asn residues that possibly participate in glycosylation (marked in red) in hClC-6 are situated in a region that is poorly conserved among the other members of the CLC family and located between predicted helices K and M. (C) Western blots of membrane fractions of COS-1 cells expressing WT or mutant hClC-6. (a), Glycosylation pattern of the triple (AAA: N410A/N422A/N432A) and quadruple (AAAA: N137A/N410A/N422A/N432A) mutant compared to WT and WT with tunicamycin. (b), Glycosylation pattern of the double mutants (AAN: N410A/N422A; ANA: N10A/N432A; NAA: N422A/N432A) compared to glycosylated WT and the triple mutant (AAA). (c), Glycosylation pattern of the single mutants (ANN: N410A, NAN: N422A, NNA: N432A) compared to the glycosylated WT and deglycosylated WT treated with tunicamycin. (d), Effect of PNGaseF treatment on a single (ANN: N410A) and double (AAN: N410A/N422A) mutant compared to WT and triple (AAA) mutant. (e), Effect of EndoH treatment on a single (ANN: N410A) and double (AAN: N410A/N422A) mutant compared to WT and triple (AAA) mutant. Bands marked with an asterisk, occasionally observed in the WT protein and frequently observed in single mutants after sustained exposure represent possible intermediary biosynthetic products, which are PNGaseF-sensitive and EndoH-insensitive as shown in panels (d) and (e).</p

Grade point average over different examination moments and master.

Grade point average over different examination moments and master.</p