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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.
MICROSTRUCTURE AND NOISE CHARACTERISTICS IN Co - Cr BASED ALLOY THIN FILM MAGNETIC RECORDING MEDIA
Y. Koshimoto,S. Hirono,T. Ohkubo,S. Umemura,Y. Maeda 한국자기학회 1995 韓國磁氣學會誌 Vol.5 No.5
Reducing media noise is a key to realizing high areal recording density. However, perpendicularly oriented Co-Cr films, strong candidates for high density recording media, have received little attention in terms of recording noise. We studied the noise characteristics of Co-Cr based alloy films by varying the compositionally separated structure.<br/> Co-Cr films with fine compositionally separated microstructure were observed to have low noise, which recording density dependence were negative or week. Studies on control of the microstructure and magnetic properties in combination with noise analysis are important to realize high density recording media.
Comparative Sequence Analysis of a Multidrug-Resistant Plasmid from Aeromonas hydrophila
del Castillo, Carmelo S.,Hikima, Jun-ichi,Jang, Ho-Bin,Nho, Seong-Won,Jung, Tae-Sung,Wongtavatchai, Janenuj,Kondo, Hidehiro,Hirono, Ikuo,Takeyama, Haruko,Aoki, Takashi American Society for Microbiology 2013 Antimicrobial agents and chemotherapy Vol.57 No.1
<B>ABSTRACT</B><P>Aeromonas hydrophilais a pathogenic bacterium that has been implicated in fish, animal, and human disease. Recently, a multidrug resistance (MDR) plasmid, pR148, was isolated fromA. hydrophilaobtained from a tilapia (Oreochromis niloticus) farm in Thailand. pR148 is a 165,906-bp circular plasmid containing 147 coding regions showing highest similarity to pNDM-1_Dok1, an MDR plasmid isolated from a human pathogen. pR148 was also very similar to other IncA/C plasmids isolated from humans, animals, food, and fish. pR148 contains a mercuric resistance operon and encodes the complete set of genes for the type 4 secretion system. pR148 encodes a Tn<I>21</I>type transposon. This transposon contains the drug resistance genes<I>qacH</I>,<I>bla</I>OXA-10,<I>aadA1</I>, and<I>sul1</I>in a class 1 integron;<I>tetA</I>and<I>tetR</I>in transposon Tn<I>1721</I>; and<I>catA2</I>and a duplicate<I>sul1</I>in a locus showing 100% similarity to IncU plasmids isolated from fish. The<I>bla</I>OXA-10and<I>aadA1</I>genes showed 100% similarity to those from theAcinetobacter baumanniiAYE genome. The similarity of pR148 to a human pathogen-derived plasmid indicates that the plasmids were either transferred between different genera or that they are derived from a common origin. Previous studies have shown that IncA/C plasmids retain a conserved backbone, while the accessory region points to lateral gene transfer. These observations point out the dangers of indiscriminate use of antibiotics in humans and in animals and the necessity of understanding how drug resistance determinants are disseminated and transferred.</P>
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.
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.
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.