Improved Culture-Based Isolation of Differentiating Endothelial Progenitor Cells from Mouse Bone Marrow Mononuclear Cells

Numerous endothelial progenitor cell (EPC)-related investigations have been performed in mouse experiments. However, defined characteristics of mouse cultured EPC have not been examined. We focused on fast versus slow adherent cell population in bone marrow mononuclear cells (BMMNCs) in culture and examined their characteristics. After 24 h-culture of BMMNCs, attached (AT) cells and floating (FL) cells were further cultured in endothelial differentiation medium separately. Immunological and molecular analyses exhibited more endothelial-like and less monocyte/macrophage-like characteristics in FL cells compared with AT cells. FL cells formed thick/stable tube and hypoxia or shear stress overload further enhanced these endothelial-like features with increased angiogenic cytokine/growth factor mRNA expressions. Finally, FL cells exhibited therapeutic potential in a mouse myocardial infarction model showing the specific local recruitment to ischemic border zone and tissue preservation. These findings suggest that slow adherent (FL) but not fast attached (AT) BMMNCs in culture are EPC-rich population in mouse

Cultivation of Research Literacy in the First Year of Graduate School: A Report on the Educational Practices of "Introduction to International Cultural Studies" in the Master's Course of the Graduate School of International Cultural Studies

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Fgf16 is required for specification of GABAergic neurons and oligodendrocytes in the zebrafish forebrain.

Fibroblast growth factor (Fgf) signaling plays crucial roles in various developmental processes including those in the brain. We examined the role of Fgf16 in the formation of the zebrafish brain. The knockdown of fgf16 decreased cell proliferation in the forebrain and midbrain. fgf16 was also essential for development of the ventral telencephalon and diencephalon, whereas fgf16 was not required for dorsoventral patterning in the midbrain. fgf16 was additionally required for the specification and differentiation of γ-aminobutyric acid (GABA)ergic interneurons and oligodendrocytes, but not for those of glutamatergic neurons in the forebrain. Cross talk between Fgf and Hedgehog (Hh) signaling was critical for the specification of GABAergic interneurons and oligodendrocytes. The expression of fgf16 in the forebrain was down-regulated by the inhibition of Hh and Fgf19 signaling, but not by that of Fgf3/Fgf8 signaling. The fgf16 morphant phenotype was similar to that of the fgf19 morphant and embryos blocked Hh signaling. The results of the present study indicate that Fgf16 signaling, which is regulated by the downstream pathways of Hh-Fgf19 in the forebrain, is involved in forebrain development

A new hierarchically porous Pd@HSQ monolithic catalyst for Mizoroki–Heck cross-coupling reactions

Pore architecture of catalyst supports is an important factor facilitating accessibility of reactants to catalytic sites. This holds the key to improving catalytic activities. Amongst various catalytic reactions, supported Pd nanoparticles-catalyzed C–C cross-coupling reactions have been attracting a great deal of attention in the last decade. Although various supports have been examined, applications of hierarchically porous monolithic materials have never been reported, mainly because of difficulties in multistep synthesis of catalysts. We herein report a novel on-site reduction-based methodology using hierarchically porous hydrogen silsesquioxane (HSQ) monoliths for one-step synthesis of Pd nanoparticles-embedded monoliths (Pd@HSQ). Characterization of these monoliths evidences the on-site reduction, i.e. formation of Pd nanoparticles and conversion of Si–H present in the monolith to Si–O . Fast, quantitative reduction of Pd[2+] to Pd(0) to form supported Pd nanoparticles is achieved with preservation of the porous structure of the original monolith, which makes this material attractive as a catalyst for C–C cross-coupling reactions. The obtained Pd@HSQ catalyst has been employed in the Mizoroki–Heck cross-coupling reaction. High accessibility of reactant molecules, undetectable leaching of Pd nanoparticles and easy separation of the monolith from liquid media provide high catalytic activity, reusability and easy handling

Overexpression of the protein disulfide isomerase AtCYO1 in chloroplasts slows dark-induced senescence in Arabidopsis

Abstract Background Chlorophyll breakdown is the most obvious sign of leaf senescence. The chlorophyll catabolism pathway and the associated proteins/genes have been identified in considerable detail by genetic approaches combined with stay-green phenotyping. Arabidopsis CYO1 (AtCYO1), a protein disulfide reductase/isomerase localized in the thylakoid membrane, is hypothesized to assemble the photosystem by interacting with cysteine residues of the subunits. Results In this study, we report that ectopic overexpression of AtCYO1 in leaves induces a stay-green phenotype during darkness, where oxidative conditions favor catabolism. In AtCYO1ox leaves, Fv/Fm and both chlorophyll a and chlorophyll b content remained high during dark-induced senescence. The thylakoid ultrastructure was preserved for a longer time in AtCYO1ox leaves than in wild type leaves. AtCYO1ox leaves maintained thylakoid chlorophyll-binding proteins associated with both PSII (D1, D2, CP43, CP47, LHCB2, and Cyt f) and PSI (PSA-A/B), as well as stromal proteins (Rubisco and ferredoxin-NADP+ reductase). AtCYO1ox did not affect senescence-inducible gene expression for chlorophyll catabolism or accumulation of chlorophyll catabolites. Conclusions Our results suggest that ectopic overexpression of AtCYO1 had a negative impact on the initiation of chlorophyll degradation and proteolysis within chloroplasts. Our findings cast new light on the redox regulation of protein disulfide bonds for the maintenance of functional chloroplasts

Comparison of cell proliferation and cell death patterns in control embryos and <i>fgf16</i> morphants.

<p>(A, B) Control embryos (A) and embryos injected with <i>fgf16</i> MO (B) were stained using an anti-H3P antibody. Panels show representative horizontal sections of the head region at 24 hpf. (C, D) The percentage of proliferating cells labelled with the anti-pH3 antibody in the forebrain (C) and midbrain (D) of control embryos and embryos injected with <i>fgf16</i> MO. Results are the mean ± S.D. for three independent sections from three embryos. The significance of differences in mean values was assessed with the Student’s <i>t</i>-test. Asterisks indicate significant differences from the control (*<i>P</i><0.05). The forebrain (fb) and midbrain (mb) regions, which we defined in the sections, are separated by black lines. Scale bar: 25 µm.</p

Gene expression in the midbrain and MHB of the <i>fgf16</i> morphants.

<p>The expression of <i>fgf8</i> (A, B), <i>otx2</i> (C, D), <i>pax7a</i> (E, F), and <i>nkx6.2</i> (G, H) in wild-type embryos (A, C, E, G) and <i>fgf16</i> morphants (B, D, F, H) at 24 hpf. Lateral views with anterior to the left and dorsal to the top.</p

Interactions between <i>fgf16</i> and Hh signaling in the forebrain and midbrain.

<p>The expression of <i>fgf16</i> at 16 (A, B) and 24 (C, D) hpf in wild-type embryos treated with 0.95% ethanol (A, C) or cyclopamine (B, D). Arrows in panels A and C indicate <i>fgf16</i> expression in the telencephalon. The arrowhead in panel C indicates <i>fgf16</i> expression in the midbrain. Lateral views with anterior to the left and dorsal to the top.</p

Expression pattern of <i>fgf16</i> in the brain during zebrafish embryonic development.

<p>(A–D) Expression pattern of <i>fgf16</i> in zebrafish embryos at the indicated stages as detected by whole-mount <i>in</i><i>situ</i> hybridization. Lateral views with anterior to the left and dorsal to the top. Arrows and arrowheads indicate the pituitary gland and epiphysis, respectively. di, diencephalon; mb, midbrain; te, telencephalon.</p