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Ohtani, M.,Hikima, J.i.,Kondo, H.,Hirono, I.,Jung, T.S.,Aoki, T. Pergamon Press ; Elsevier Science Ltd ; Pergamon 2011 Developmental and comparative immunology Vol.35 No.5
Cytosolic pattern recognition receptors such as retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) play an important role in sensing viral RNAs. The receptor encoded by melanoma differentiation-associated gene 5 (MDA5), an RLR, recognizes viral RNA in the cytoplasm and enhances antiviral response in host cells. The full-length MDA5 gene in Japanese flounder, Paralichthys olivaceus was cloned and found to have 11,251 nucleotides. MDA5 transcript abundance was significantly increased in whole kidney infected with viral hemorrhagic septicemia virus (VHSV) as well as whole kidney and peripheral blood leukocytes stimulated with poly I:C in vitro. Hirame natural embryo (HINAE) cells overexpressing MDA5 showed a lower cytopathic effect (CPE) against VHSV, hirame rhabdovirus (HIRRV) and infectious pancreatic necrosis virus (IPNV) infection. When infected with VHSV, MDA5-overexpressing HINAE cells had 24-75 fold lower virus titer than normal HINAE cells. These results suggest that Japanese flounder MDA5 is involved in the induction of antiviral response.
Hikima, J.i.,Ohtani, M.,Kondo, H.,Hirono, I.,Jung, T.S.,Aoki, T. Pergamon Press ; Elsevier Science 2011 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol.35 No.3
Both PU.1 and C/EBPα transcription factors play important roles in myeloid development and inflammatory response. These transcripts were cloned from the Japanese flounder (Paralichthys olivaceus) and were highly conserved with those of other vertebrates. PU.1 mRNA was mainly expressed in lymphoid tissues while C/EBPα mRNA was widely expressed in all tissues examined. Higher levels of PU.1 mRNA were expressed in the IgM<SUP>+</SUP> cells of both PBL and KL, while C/EBPα expression was higher only in the IgM<SUP>-</SUP> cells of KL. The expression of C/EBPα mRNA was induced only in KL stimulated with LPS. Interestingly, PU.1 mRNA expression was induced by Edwardsiella tarda, whereas the expression of C/EBPα mRNA was induced by Streptococcus iniae infection. Both PU.1 and C/EBPα drove transcription from the LPS-responsive region of the Japanese flounder TNFα gene, suggesting that both PU.1 and C/EBPα induced by bacterial infection are involved in inflammation mediated through TNFα expression.
InGaAs quantum dot molecules during selective etching using an In droplet mask
Lee, Jihoon,Wang, Zhiming,Hirono, Yusuke,Kim, Eun-Soo,Koo, Sang-Mo,Dorogan, Vitaliy G,Mazur, Yuriy I,Song, Sangmin,Park, Gamyoung,Salamo, Gregory J Institute of Physics [etc.] 2011 Journal of Physics. D, Applied Physics Vol.44 No.2
<P>We investigated the optical transition of InGaAs quantum dot molecules (QDMs) during selective etching of GaAs using In droplets to demonstrate low-density QDMs. During the selective etching, In droplets act as nanoscale masks and only QDMs underneath the droplets survive, by which process low-density QDMs are fabricated. The thickness of selective GaAs etching is systematically varied and a gradual red-shift is observed with the increased etching thickness. The continuing red-shift can be explained by the strain relaxation due to GaAs etching. This technique to achieve low-density QDMs by selective etching using droplets as nanoscale mask is a simple and flexible approach. This study can find applications in single QDM spectroscopy and other spectroscopic techniques.</P>
Ohtani, M.,Hikima, J.i.,Jung, T.S.,Kondo, H.,Hirono, I.,Takeyama, H.,Aoki, T. Academic Press 2013 FISH AND SHELLFISH IMMUNOLOGY Vol.34 No.2
Phage display libraries are used to screen for nucleotide sequences that encode immunoglobulin variable (V) regions that are specific for a target antigen. We previously constructed an immunoglobulin new antigen receptor (IgNAR) phage display library. Here we used this library to obtain an IgNAR V region that is specific for viral hemorrhagic septicemia virus (VHSV). A phage clone (clone 653) was found to be specific for VHSV by the biopanning method. The V region of clone 653 was used to construct a 6 x His tagged recombinant IgNAR-653 V protein (rIgNAR-653) using the Escherichia coli pET system. The rIgNAR-653 protein bound specifically to VHSV, confirming its activity.
Low-Density Quantum Dot Molecules by Selective Etching Using in Droplet as a Mask
Jihoon Lee,Wang, Z M,Hirono, Y,Dorogan, V G,Mazur, Y I,Eun-Soo Kim,Sang-Mo Koo,Seunghyun Park,Sangmin Song,Salamo, G J IEEE 2011 IEEE TRANSACTIONS ON NANOTECHNOLOGY Vol.10 No.3
<P>We demonstrate low-density quantum dot molecules (QDMs) by selective etching using In droplets as a mask. Selective etching is performed with InGaAs QDMs buried underneath GaAs capping layer, on which In droplets are formed by droplet epitaxy using molecular beam epitaxy. During the chemical etching, the droplets act as a mask and QDMs underneath the droplets that only survive. Photoluminescence measurement from the selectively etched QDMs in mesa structures shows a much reduced intensity, which indicates low-density QDMs. This technique provides a simple and flexible method to attain low-density QDMs. The density can be easily modified by the control of the size and density of In droplets, which is suitable for single QDM spectroscopy and for their device applications.</P>
Generation of monoclonal antibodies specific for ORF68 of koi herpesvirus
Aoki, T.,Takano, T.,Unajak, S.,Takagi, M.,Kim, Y.R.,Park, S.B.,Kondo, H.,Hirono, I.,Saito-Taki, T.,Hikima, J.i.,Jung, T.S. Pergamon Press 2011 Comparative immunology, microbiology and infectiou Vol.34 No.3
Outbreaks of koi herpesvirus (KHV) infection in carp are still a serious problem worldwide. KHV is closely related to other two cyprinid herpesviruses, pox herpesvirus (CHV) and haematopoietic necrosis herpesvirus (CyHV-2) in goldfish. In this study, two major KHV antigenic proteins (ORF62 and ORF68) were identified by immunoscreening using a KHV-specific polyclonal antibody, and then monoclonal antibodies were generated for immunodiagnostic studies. After screening hybridoma cells, one mAb against ORF68 (mAb-7C6) was obtained but no mAbs against ORF62. mAb-7C6 specifically reacted with a lysate of KHV-infected koi fin cells (KF-1 cells) but not with lysates of CHV- or CyHV-2-infected KF-1 cells in an immuno-blotting analysis. Similar results were shown in the following tests: (1) a indirect fluorescent antibody test using infected KF-1 cells and (2) an immunohistochemical investigation by fast red stain (infected liver) or FITC detection (infected spleen). These results suggested that mAb-7C6 specifically reacts with KHV ORF68 protein.