TbGT8 is a bifunctional glycosyltransferase that elaborates<em> N</em>-linked glycans on a protein phosphatase AcP115 and a GPI-anchor modifying glycan in <em>Trypanosoma brucei</em>

AbstractThe procyclic form of Trypanosoma brucei expresses procyclin surface glycoproteins with unusual glycosylphosphatidylinositol-anchor side chain structures that contain branched N-acetyllactosamine and lacto-N-biose units. The glycosyltransferase TbGT8 is involved in the synthesis of the branched side chain through its UDP-GlcNAc: βGal β1-3N-acetylglucosaminyltransferase activity. Here, we explored the role of TbGT8 in the mammalian bloodstream form of the parasite with a tetracycline-inducible conditional null T. brucei mutant for TbGT8. Under non-permissive conditions, the mutant showed significantly reduced binding to tomato lectin, which recognizes poly-N-acetyllactosamine-containing glycans. Lectin pull-down assays revealed differences between the wild type and TbGT8 null-mutant T. brucei, notably the absence of a broad protein band with an approximate molecular weight of 110kDa in the mutant lysate. Proteomic analysis revealed that the band contained several glycoproteins, including the acidic ecto-protein phosphatase AcP115, a stage-specific glycoprotein in the bloodstream form of T. brucei. Western blotting with an anti-AcP115 antibody revealed that AcP115 was approximately 10kDa smaller in the mutant. Enzymatic de-N-glycosylation demonstrated that the underlying protein cores were the same, suggesting that the 10-kDa difference was due to differences in N-linked glycans. Immunofluorescence microscopy revealed the colocalization of hemagglutinin epitope-tagged TbGT8 and the Golgi-associated protein GRASP. These data suggest that TbGT8 is involved in the construction of complex poly-N-acetyllactosamine-containing type N-linked and GPI-linked glycans in the Golgi of the bloodstream and procyclic parasite forms, respectively

Complement C9 expression is associated with damaged myocardial cells in pediatric sudden death cases of fulminant myocarditis.

Background:Because disease progression is so fast in sudden death of acute fulminant myocarditis, damage of myocardial cells is not evident in routine hematoxylin and eosin staining. To understand damage to myocardial cells and the mechanism of sudden death, immunohistochemical staining was performed for two forensic autopsy cases.Case presentation:The patients were a healthy 5-year-old girl and 8-year-old boy. They suddenly died within 2 days of appearance of flu-like symptoms. An autopsy showed accumulation of yellowish-clear pericardial fluid containing fibrin deposits, fluid blood in the heart, and congestion of visceral organs. Histologically, minor necrosis or degeneration of myocardial cells with mainly lymphocytic infiltration was observed sometimes in tissue sections. Immunohistochemically, positive complement C9 staining and negative sirtuin 1 staining were found. These findings suggested wide damage of myocardial cells, even in regions with no marked changes in myocardial cells with hematoxylin and eosin staining. These areas corresponded to those with strong accumulation of lymphocytes.Conclusions:Immunohistochemistry for complement C9 and sirtuin 1 might become a new tool for evaluating damage of myocardial cells of fulminant acute myocarditis

BRASSICA NAPUSヒンシュ TOPAS ニ オケル レンゾクテキハイケイセイ オヨビ ニジハイ カラ ノ ショクブツタイサイセイケイ

Brassica napus品種`Topas\u27における連続的な二次胚形成及び植物体再生系の確立を試みた。熱ショックによって未熟種子胚より誘導した二次胚を,植物成長調節物質無添加のB5培地へ継代した。培養後70%の二次胚において,徒長した胚軸の表面に次代の二次胚が直接形成された。これら二次胚を同様に継代培養した結果,約70%の胚より新たな二次胚が形成され,次胚形成能が継代培養を通じて維持されていたことが示された。二次胚に10μMアブシジン酸(ABA)処理をした結果,胚は乾燥耐性を獲得し,60%の二次胚が乾燥後においても生育しその全てが正常に再生した。一方,ABA処理後,乾燥処理を行わなかった胚は胚軸が徒長する異常な生育を示し,ABA無処理胚は乾燥処理によって全て枯死した。A method for continuous secondary embryo formation and plant regeneration in Brassica napus cv. `Topas\u27 is described. Secondary embryos that emerged from immature zygotic embryos via heat shock were separated and subcultured onto B5 plant growth regulator-free medium. Most secondary embryos (i.e. 70%) produced secondary embryos in a subsequent generation directly on the surface of elongated hypocotyls. Similarly, about 70% of the secondary embryos formed newly produced secondary embryos in a subsequent generation after subculturing, and the embryogenic potential of these secondary embryos has been maintained by repetitive subculture. The application of 10μM abscisic acid (ABA) induced desiccation tolerance in secondary embryos. Consequently, 60% of the desiccated embryos grew, all of which regenerated into normal plants. On the other hand, ABA-treated secondary embryos without desiccation treatment grew abnormally having an elongated hypocotyl, and all secondary embryos not treated with ABA lost their viability after desiccation

Enhancing the Therapeutic Efficacy of Bone Marrow-Derived Mononuclear Cells with Growth Factor-Expressing Mesenchymal Stem Cells for ALS in Mice.

Several treatments have been attempted in amyotrophic lateral sclerosis (ALS) animal models and patients. Recently, transplantation of bone marrow-derived mononuclear cells (MNCs) was investigated as a regenerative therapy for ALS, but satisfactory treatments remain to be established. To develop an effective treatment, we focused on mesenchymal stem cells (MSCs) expressing hepatocyte growth factor, glial cell line-derived neurotrophic factor, and insulin-like growth factor using human artificial chromosome vector (HAC-MSCs). Here, we demonstrated the transplantation of MNCs with HAC-MSCs in ALS mice. As per our results, the progression of motor dysfunction was significantly delayed, and their survival was prolonged dramatically. Additional analysis revealed preservation of motor neurons, suppression of gliosis, engraftment of numerous MNCs, and elevated chemotaxis-related cytokines in the spinal cord of treated mice. Therefore, growth factor-expressing MSCs enhance the therapeutic effects of bone marrow-derived MNCs for ALS and have a high potential as a novel cell therapy for patients with ALS

Monolithic electrode for electric double-layer capacitors based on macro/meso/microporous S-Containing activated carbon with high surface area

Macro/meso/microporous carbon monoliths doped with sulfur have been prepared from sulfonated poly(divinylbenzene) networks followed by the activation with CO_2 resulted in the activated carbon monoliths with high surface area of 2400 m^2 g^[−1]. The monolithic electrode of the activated carbon shows remarkably high specific capacitance (175 F g^[−1] at 5 mV_s^[−1] and 206F_g^[−1] at 0.5 Ag^[−1])