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- 저자 : Nakasaki, Manando (검색결과 5 건)
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Defective natural killer cell activity in a mouse model of eczema herpeticum
Kawakami, Yuko,Ando, Tomoaki,Lee, Jong-Rok,Kim, Gisen,Kawakami, Yu,Nakasaki, Tae,Nakasaki, Manando,Matsumoto, Kenji,Choi, Youn Soo,Kawakami, Toshiaki Elsevier 2017 The journal of allergy and clinical immunology Vol.139 No.3
<P><B>Background</B></P> <P>Patients with atopic dermatitis (AD) are susceptible to several viruses, including herpes simplex virus (HSV). Some patients experience 1 or more episodes of a severe skin infection caused by HSV termed eczema herpeticum (EH). There are numerous mouse models of AD, but no established model exists for EH.</P> <P><B>Objective</B></P> <P>We sought to establish and characterize a mouse model of EH.</P> <P><B>Methods</B></P> <P>We infected AD-like skin lesions with HSV1 to induce severe skin lesions in a dermatitis-prone mouse strain of NC/Nga. Gene expression was investigated by using a microarray and quantitative PCR; antibody titers were measured by means of ELISA; and natural killer (NK) cell, cytotoxic T-cell, regulatory T-cell, and follicular helper T-cell populations were evaluated by using flow cytometry. The role of NK cells in HSV1-induced development of severe skin lesions was examined by means of depletion and adoptive transfer.</P> <P><B>Results</B></P> <P>Inoculation of HSV1 induced severe erosive skin lesions in eczematous mice, which had an impaired skin barrier, but milder lesions in small numbers of normal mice. Eczematous mice exhibited lower NK cell activity but similar cytotoxic T-cell activity and humoral immune responses compared with normal mice. The role of NK cells in controlling HSV1-induced skin lesions was demonstrated by experiments depleting or transferring NK cells.</P> <P><B>Conclusion</B></P> <P>A murine model of EH with an impaired skin barrier was established in this study. We demonstrated a critical role of defective NK activities in the development of HSV1-induced severe skin lesions in eczematous mice.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
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Sang Hagk Kwon,Kiyohiko Nakasaki 한국공업화학회 2015 Journal of Industrial and Engineering Chemistry Vol.21 No.1
Synthetic peptone wastewater was used as a substrate in a laboratory-scale methane fermentationreactor. Methane production ceased when the hydraulic retention time (HRT) was changed from 4 d to1 d. A DNA-based method revealed that the amount of acidogenic bacteria increased, whilemethanogenic archaea decreased under these conditions. The short HRT resulted in a high concentrationof dissolved oxygen in the wastewater, which adversely affected the archaea by stimulating bacterialproduction of high concentrations of volatile fatty acids and ammonium ion, and growth of thebacterium belonging to Desulfoglaeba that competes with methanogens for reducing power.
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( Osamu Ariga ),( Naoki Okamoto ),( Naomi Harimoto ),( Kiyohiko Nakasaki ) 한국미생물 · 생명공학회 2014 Journal of microbiology and biotechnology Vol.24 No.1
α-Neoagarooligosaccharide (α-NAOS) hydrolase was purified from Cellvibrio sp. OA-2007 by using chromatographic techniques after hydroxyapatite adsorption. The molecular masses of α-NAOS hydrolase estimated using SDS-PAGE and gel filtration chromatography were 40 and 93 kDa, respectively, and the optimal temperature and pH for the enzyme activity were 32ºC and 7.0-7.2. α-NAOS hydrolase lost 43% of its original activity when incubated at 35ºC for 30 min. The enzyme hydrolyzed neoagarobiose, neoagarotetraose, and neoagarohexaose to galactose, agarotriose, and agaropentaose, respectively, and produced 3,6-anhydro-L-galactose concomitantly; however, it did not degrade agarose.
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Cloning of Agarase Gene from Non-Marine Agarolytic Bacterium Cellvibrio sp
( Ariga Osamu ),( Takayoshi Inoue ),( Hajime Kubo ),( Kimi Minami ),( Mitsuteru Nakamura ),( Michi Iwai ),( Hironori Moriyama ),( Mitsunori Yanagisawa ),( Kiyohiko Nakasaki ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.9
Agarase genes of non-marine agarolytic bacterium Cellvibrio sp. were cloned into Escherichia coli and one of the genes obtained using HindIII was sequenced. From nucleotide and putative amino acid sequences (713 aa, molecular mass; 78,771 Da) of the gene, designated as agarase AgaA, the gene was found to have closest homology to the Saccharophagus degradans (formerly, Microbulbifer degradans) 2-40 aga86 gene, belonging to glycoside hydrolase family 86 (GH86). The putative protein appears to be a non-secreted protein because of the absence of a signal sequence. The recombinant protein was purified with anion exchange and gel filtration columns after ammonium sulfate precipitation and the molecular mass (79 kDa) determined by SDS-PAGE and subsequent enzymography agreed with the estimated value, suggesting that the enzyme is monomeric. The optimal pH and temperature for enzymatic hydrolysis of agarose were 6.5 and 42.5ºC, and the enzyme was stable under 40ºC. LC-MS and NMR analyses revealed production of a neoagarobiose and a neoagarotetraose with a small amount of a neoagarohexaose during hydrolysis of agarose, indicating that the enzyme is a β-agarase.
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