bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Influence of Ovine MHC Class IIDRB1Alleles on Immune Response in Bovine Leukemia Virus Infection
Authors: Konnai Satoru , Takeshima Shin‐nosuke , Tajima Shigeru , Yin Shan Ai , Okada Kosuke , Onuma Misao , Aida Yoko 
Published: November 2013

AbstractWe have reported previously that the alleles of the ovine leukocyte antigen (OLA)‐DRB1gene that encode the Arg‐Lys (RK) motif and the Ser‐Arg (SR) motif at positions β70/71of the OLA‐DRβ1 domain are associated with resistance and susceptibility, respectively, to development of bovine leukemia virus (BLV)‐induced ovine lymphoma. Here, to investigate the different immune response in sheep that carried alleles associated with resistance and susceptible for 30 weeks after infection with BLV, we selected sheep that had the RK/RK or SR/SR genotype among the 52 sheep analyzed by polymerase chain reaction‐restriction fragment length polymorphism and DNA sequencing of PCR product for theOLA‐DRB1exon 2 and infected them with BLV. Although the number of BLV‐infected cells and virus titer had been maintaining low levels throughout the experimental period, the sheep with the RK/RK genotype could induce expansion of CD5−B‐cells and rapid production of neutralizing antibody in the early phase of infection. The level of incorporation of [3H]thymidine by peripheral blood mononuclear cells from the sheep with RK/RK genotype gave a strong response to BLV virion antigen and synthetic antigenic peptides that corresponded to T‐helper epitope of the BLV envelope glycoprotein gp51. In contrast, the sheep with SR/SR genotype showed a strong response to BLV virion antigen and synthetic antigenic peptides that corresponded to T‐cytotoxic and B‐cell epitopes. In such cases, the animals with the RK/RK strongly expressed IFN‐7, the animals with SR/SR genotype strongly expressed IL‐2. To determine the proliferating cells, we tried a blocking assay with monoclonal antibodies such as anti‐CD4, ‐CD8 and ‐DR molecule. We found that these proliferating cells were MHC‐restricted CD4+T‐cells.
Comparison of a Multiplex‐PCR Assay with Mycolic Acids Analysis and Conventional Methods for the Identification of Mycobacteria
Authors: Tanaka Ioshie Ibara , Anno Ivone Shizuko , de Andrade Leite Sergio Roberto , Cooksey Robert C. , Leite Clarice Queico Fujimura 
Published: November 2013

AbstractA fast, sensitive and cost‐effective multiplex‐PCR assay for Mycobacterium tuberculosis complex (MTC) and Mycobacterium avium (M. avium) identification for routine diagnosis was evaluated. A total of 158 isolates of mycobacteria from 448 clinical specimens from patients with symptoms of mycobacterial disease were analyzed. By conventional biochemical methods 151 isolates were identified as M. tuberculosis, five as M. avium and two as Mycobacterium chelonae (M. chelonae). Mycolic acid patterns confirmed these results. Multiplex‐PCR detected only IS6110 in isolates identified as MTC, and IS1245 was found only in the M. avium isolates. The method applied to isolates from two patients, identified by conventional methods and mycolic acid analysis, one as M. avium and other as M. chelonae, resulted positive for IS6110, suggesting co‐infection with M. tuberculosis. These patients were successfully submitted to tuberculosis treatment. The multiplex‐PCR method may offer expeditious identification of MTC and M. avium, which may minimize risks for active transmission of these organisms and provide useful treatment information.
Analysis of the Envelope Proteins of Heat‐Shocked Vibrio parahaemolyticus Cells by Immunoblotting and Biotin‐Labeling Methods
Authors: Wong Hin‐chung , Chen Yu‐Chih 
Published: November 2013

AbstractVibrio parahaemolyticus, a common enteropathogen in tropical and subtropical coastal regions, exhibits significant adaptive acid tolerance response and heat‐shock response, and the envelope proteins induced by stresses are suggested to be associated with virulence. This work examined the heat‐shock proteins located in the envelope of V. parahaemolyticus by two rapid methods; namely, the immunoblotting and biotin‐labeling methods. The bacterial cells were cultured at 25 C and heat shocked at 37 or 42 C for 1 or 2 hr. The cells were first lysed, then proteins were separated by gel electrophoresis and probed with antiserum raised against heat‐shocked cells. Next, the heat‐shocked cells were examined by labeling with water soluble sulfo‐NHS‐LC‐biotin. Proteins of 33, 61, 66, 71, 78, 92 and 101 kDa were induced, while 55, 86, 102, 120 and 160 kDa proteins were markedly enhanced in the envelope of the heat‐shocked V. parahaemolyticus cells. The biotin tagged envelope proteins were purified using a monomeric avidin column, and the N‐terminal sequence was determined and compared with other high identity protein sequences. The sequence results suggest that Vph1 (55 kDa), Vph2 (46 kDa) and Vph3 (42 kDa) are de novo synthesized heat‐shock proteins located in the envelope of this pathogen, and the functions of these proteins in stress protection and virulence have yet to be determined.
Mycobacterial Infection in TLR2 and TLR6 Knockout Mice
Authors: Sugawara Isamu , Yamada Hiroyuki , Li Chuanyou , Mizuno Satoru , Takeuchi Osamu , Akira Shizuo 
Published: November 2013

AbstractTo investigate the role of TLR in the development of murine tuberculosis in vivo, TLR2 and TLR6 knockout (KO) mice were infected with Mycobacterium tuberculosis by placing them in the exposure chamber of an airborne infection apparatus. Both TLR2 and TLR6 KO mice survived until sacrifice at 12 weeks after infection. Infected TLR2 KO mice developed granulomatous pulmonary lesions with neutrophil infiltration, which were slightly larger in size than those in wild‐type mice. Pulmonary levels of the mRNAs for inducible nitric oxide synthase (iNOS), TNF‐α, TGF‐β, IL‐1β, and IL‐2 were significantly lower, but levels of the mRNAs for IL‐4 and IL‐6 were higher, than in wild‐type (WT) mice. No significant difference was recognized in cytokine mRNA expression between TLR2 KO and WT mice at 12 weeks after infection. DNA binding by NF‐κB was low in TLR2 KO mice. On the other hand, TLR6 KO mice were not different from WT mice in terms of pulmonary histopathology, mRNA expression and CFU assay. Therefore, TLR2 does not play an essential role in the pathogenesis of murine tuberculosis, although it is important for defense against mycobacterial infection.
Further Characterization of the Rabies Virus Glycoproteins Produced by Virus‐Infected and G cDNA‐Transfected Cells Using a Monoclonal Antibody, #1‐30‐44, Which Recognizes an Acid‐Sensitive Epitope
Authors: Kankanamge Pushpa Jenette , Irie Takashi , Shoji Jun'ichi , Tochikura Tadafumi S. , Kawai Akihiko 
Published: November 2013

AbstractExpression of rabies virus glycoprotein (G) by G cDNA‐transfected mammalian cells resulted in the production of only a fusion‐negative form. Low pH‐dependent fusion activity, however, was seen when the expression was done under control of the T7 promoter with the help of recombinant vaccinia virus (RVV‐T7) that provided T7 RNA polymerase. Fusion‐inactive G proteins were transported to the cell surface as being detected by a conformational epitope‐specific monoclonal antibody (mAb; #1‐46‐12). The fusion‐inactive G proteins were recognized by most of our 13 conformation‐specific mAbs, except for one mAb, #1‐30‐44, that recognized the low pH‐sensitive conformational epitope. When the G gene expression was done with the help of RVV‐T7, although most G proteins remained in the epitope‐negative form, a small fraction of G gene products were 1‐30‐44 epitope‐positive, and cell fusion activity could be seen when cells were exposed to low pH conditions. From these results, we conclude that acquisition of low pH‐dependent fusion activity is closely related to structural maturation of the G protein to form the low pH‐sensitive 1‐30‐44 epitope. Such maturation seems to be dependent on certain rabies virus‐induced cellular conditions or functions, which might also be provided in part by the vaccinia virus infection. We further assume that expression of G cDNA alone mostly results in the production of mis‐folded and/or differently folded forms of G protein, and only a small fraction is correctly folded even under RVV‐T7‐mediated expression conditions.
T‐Cell Receptor Variable γ Chain Gene Expression in the Interaction between Rat γδ‐Type T Cells and Heat‐Shock Protein 70‐Like Molecule
Authors: Ichinohe Takashi , Ichimiya Shingo , Kishi Akihiko , Tamura Yasuaki , Kondo Nobuhiko , Ueda Gosei , Torigoe Toshihiko , Yamaguchi Akira , Hiratsuka Hiroyoshi , Hirai Itaru , Kohama Genn‐iku , Sato Noriyuki 
Published: November 2013

AbstractWe previously reported that rat T‐cell receptor (TCR) Vδ6 of T‐cell hybridomas was preferentially involved in recognition of the cell surface‐expressed 70 kDa rat heat‐shock cognate (hsc70, a constitutively expressed member of the hsp 70 family) protein‐like molecule (#067 molecule). In the present study, we analyzed usage of the TCR Vγ family of #067‐restricted T‐cell hybridomas. Our data indicated that most of these hybridomas expressed transcripts of TCR Vγ1 and/or Vγ2. However, some of the Vγ2 transcripts were out‐of‐frame, suggesting that the TCR Vγ1 family may be important for the recognition of #067‐defined molecules. TCR Vγ1 transcripts were detected in not only #067‐restricted T‐cell hybridomas, but #067‐non restricted ones as well. However, V‐J nucleotide sequences of #067‐restricted and #067‐non restricted T‐cell hybridomas suggested that #067‐restricted T‐cell hybridomas showed limited insertion of nucleotide stretch as compared with #067‐non restricted ones. In terms of amino acids, only one amino acid was added in #067‐restricted T‐cell hybridomas, whereas two or three amino acids were added in #067‐non restricted ones. These data suggest that the heterodimer of the TCR relatively short stretch form of Vγ1 molecule and TCR Vδ6 may participate in recognition of the #067 molecule.
A Rare Appearance of Influenza A(H1N2) as a Reassortant in a Community Such as Yamagata Where A(H1N1) and A(H3N2) Co‐Circulate
Authors: Mizuta Katsumi , Katsushima Noriko , Ito Sueshi , Sanjoh Kanako , Murata Toshio , Abiko Chieko , Murayama Shoko 
Published: November 2013

AbstractTo find a new influenza subtype A(H1N2), 383 isolates identified as H1 by hemagglutination inhibition test between the 1998–1999 and 2001–2002 seasons in Yamagata, Japan, were screened by reverse transcription polymerase chain reaction. As a result, 3 strains from the 1999–2000 season were identified as possibly being A(H1N2). Although several of their clones were found to be A(H1N2), A(H1N1) and A(H3N2), we could not confirm the origin of the A(H1N2) clones without the original specimens. These results suggest that a reassortment to produce A(H1N2) does not readily occur even when A(H1N1) and A(H3N2) co‐circulate in a community such as Yamagata.
Pathogenicity of Toxoplasma gondii through B‐2 Cell‐Mediated Downregulation of Host Defense Responses
Authors: Mun Hye‐Seong , Aosai Fumie , Chen Mei , Piao Lian‐Xun , Norose Kazumi , Iwakura Yoichiro , Yano Akihiko 
Published: November 2013

AbstractIFN‐γ is the primary mediator of anti‐parasite effector mechanisms against Toxoplasma gondii. After intraperitoneal infection with the Fukaya strain of T. gondii, unirradiated IFN‐γ knock‐out (GKO) mice transferred with wild type (WT) CD8+ effector T cells from infected mice failed to induce the production of IFN‐γ and died, whereas irradiated (IR) GKO mice transferred with WT CD8+ T cells induced IFN‐γ production and survived more than 6 months. IR GKO mice transferred with WT CD8+ T cells together with GKO B‐2 cells died 8 days after infection, whereas those transferred with WT CD8+ T cells together with B‐1a or T cells survived. B‐2 cells of infected GKO mice activated CD11b+ cells for IL‐4 production, and down‐regulated NO release, STAT1 phosphorylation, and interferon regulatory factor‐1 expression in the peritoneal exudates cells of IR GKO mice transferred with WT CD8+ T cells together with GKO B‐2 cells after infection. Thus, B‐2 cells in T. gondii‐infected mice act as suppressor cells in the host defense of infected mice.
Characterization of Epstein‐Barr Virus (EBV)‐Positive NK Cells Isolated from Hydroa Vacciniforme‐Like Eruptions
Authors: Demachi Ayako , Nagata Hiroshi , Morio Tomohiro , Oyoshi Michiko K. , Zhang Yu , Tabata Nobuko , Kimura Nobuhiro , Shimizu Norio , Yamamoto Kohtaro 
Published: November 2013

AbstractRecently, the involvement of Epstein‐Barr virus (EBV) in hydroa vacciniforme (HV)‐like eruptions has been suggested. To elucidate the role of EBV in this disease, we isolated EBV‐infected cell clones from peripheral blood mononuclear cells (PBMC) and the skin lesions of a patient with HV‐like eruptions; cells isolated from PBMC were designated SNK‐12, and those from the eruption SNK‐11. Both cells expressed CD16, CD56, and HLA‐DR and had germline configurations of the T‐cell receptor and the immunoglobulin genes, indicating that the cell clones were of NK cell lineage. The analysis of EBV terminal repeats indicated that the cells were monoclonal, had identical clonality, and originated from EBV‐positive cells in the PBMC and eruption. Both clones expressed EBNA‐1, but not EBNA‐2. Although LMP‐1 was weakly detected in SNK‐11, no LMP‐1 was detected in SNK‐12. Interestingly, EBV‐infected cells required less IL‐2 for in vitro growth in the later phase of this disease and this appeared to correlate with the expression of LMP‐1, suggesting that the proliferative capacity of the EBV‐positive NK cells increased during the time course of the disease, and LMP‐1 expression might be responsible for that. This is the first report of the isolation of EBV‐infected cells from the skin lesions of HV‐like eruptions and strongly suggests that the HV‐like eruption in the patient was caused by clonal NK cells with latent EBV infection.
The Role of Non‐Chromosomal Histones in the Host Defense System
Authors: Kanai Yoshiyuki 
Published: November 2013

AbstractHistone H1 is located at the inter‐nucleosome and more correctly at both ends of the double‐stranded DNA that protrude from the nucleosome unit. It has long been recognized to be localized only inside the nuclei as a constituent for packaging nucleosome into chromatin. Thus, it could be hardly believed that detatched or solubilized histone H1 plays the role of a host defense molecule. Given the old reports on histone‐like basic proteins that show bacteriostatic functions, I herein chose some recent related articles and tried review them. Recent advances in research on the cell death mechanism makes it possible to understand that programmed cell death, (i.e. apoptosis) could serve as a good source of soluble histones. Some forms of them are highly probable to be bacteriostatic.
A Quantitative In Vitro Assay Method for Detecting Biological Activity of Endotoxin Using Prostaglandin E2 Induction in Rabbit Peripheral Blood
Authors: Ochiai Masaki , Kataoka Michiyo , Toyoizumi Hiromi , Yamamoto Akihiko , Kamachi Kazunari , Horiuchi Yoshinobu 
Published: November 2013

AbstractThe pyrogen test and the endotoxin test (the LAL test) have been playing crucial roles in detecting endotoxin in parenteral drugs. The current test methods, however, have disadvantages such as requiring a large number of animals or an inadequacy in evaluation of in vivo endotoxin activity. We attempted to establish a new assay method that can overcome the shortcomings of the current methods. We standardized the in vitro assay method by the use of prostaglandin E2 (PGE2) induction from peripheral blood of rabbits for detecting endotoxin activity. A linear dose‐response regression was attained from approximately 0.15 to 5 endotoxin units/ml of Japanese national reference standard endotoxin by the in vitro assay. The assay showed a fine correlation with the pyrogen test but not with the LAL test, when endotoxins from various bacterial sources were tested. The in vitro assay was also shown to have the capability of detecting a synergistic effect of endotoxin and parenteral drugs. The in vitro PGE2 induction test using rabbit blood was, therefore, suggested to be the appropriate test method for guaranteeing the same level of safety of parenteral drugs as the pyrogen test does.
Antigenic Analysis of Nonstructural Protein (NSP) 4 of Group A Avian Rotavirus Strain PO‐13
Authors: Borgan Mohammed Ali , Mori Yoshio , Ito Naoto , Sugiyama Makoto , Minamoto Nobuyuki 
Published: November 2013

AbstractIn order to analyze the antigenic structure of nonstructural protein (NSP) 4 of group A avian rotavirus strain PO‐13, 25 monoclonal antibodies (MAbs) against NSP4 expressed in Escherichia coli were produced. All MAbs reacted with NSP4 on Western blotting, indicating that they recognized sequential epitopes. To determine the antigenic sites (ASs) recognized by the produced MAbs, seven truncated NSP4s were expressed in E. coli. Western blotting analysis showed that there are at least four major ASs on PO‐13 NSP4, designated as AS I located in amino acids (aa) 151 to 169, AS II (aa 136 to 150), AS III (aa 112 to 133) and AS IV (aa 1 to 24). Two MAbs reacted exclusively with AS III encompassing the region that has been reported to be an enterotoxin domain. MAbs against ASs II, III and IV reacted with all avian rotaviruses tested by indirect immunofluorescent antibody assays. MAbs against AS I reacted with turkey strains, Ty‐1 and Ty‐3, but not with a chicken strain, Ch‐1. Nine of 11 MAbs against AS II cross‐reacted with NSP4 of mammalian rotavirus strains with different NSP4 genotypes. These results suggest that AS II on NSP4 is widely conserved among a variety of rotaviruses.
SATB1 Makes a Complex with p300 and Represses gp91phox Promoter Activity
Authors: Fujii Yoshito , Kumatori Atsushi , Nakamura Michio 
Published: November 2013

AbstractThe expression of gp91phox, the key component of the phagocyte NADPH oxidase, is regulated by various factors binding to its proximal promoter. Two nuclear matrix attachment region (MAR)‐binding proteins, special AT‐rich binding protein 1 (SATB1) and CCAAT displacement protein (CDP), have been reported as rare examples of gp91phox gene repressors. However, their individual roles and interactions with other factors in the promoter have not been elucidated in detail. We have focused on these two repressive proteins recognizing the bp −115 to bp −106 segment of the gene and obtained the following results: 1. SATB1 makes a complex, mainly with p300, regardless of the presence of DNA. 2. SATB1/p300 complex binding to the 5′ upstream AT‐rich region in the bp −115 to bp −106 segment represses the gp91phox promoter activity, and the repressed activity is partially released by CDP binding to the CCAAT element directly downstream of the AT‐rich region. Our findings imply a novel role for p300 in SATB1‐associated global transcription regulation.
Further Characterization of a CD99‐Related 21‐kDa Transmembrane Protein (VAP21) Expressed in Syrian Hamster Cells and Its Possible Involvement in Vesicular Stomatitis Virus Production
Authors: Tochikura Tadafumi S. , Xiao Sa , Ego Takeshi , Sagara Junji , Kawai Akihiko 
Published: November 2013

AbstractThe VAP21, a CD99‐related 21‐kDa transmembrane protein, was first detected in the enveloped virions that were grown in a Syrian hamster‐derived cell line, BHK‐21 (Sagara et al., 1997; Yamamoto et al., 1999). We further tried to elucidate the nature and properties of VAP21. The VAP21 was detected in various organs of the Syrian hamster as well as in the Syrian hamster‐derived cell lines (BHK‐21 and HmLu‐1). We could not detect the VAP21 antigen in other cell lines derived from other animal species we examined, including a Chinese hamster (CHO‐K1), mouse (neuroblastoma C1300, clone NA), dog (MDCK), monkey (COS‐7), and human (HeLa, HepG2). We tried to introduce the VAP21 gene into VAP21‐negative cell lines using a tetracycline‐regulated gene expression system. All of our trials, however, resulted in failure to establish stably positive inducible cell lines. To the contrary, we could easily establish the VAP21‐overexpressing cell lines from the Syrian hamster cell lines, which were successfully grown and maintained without any loss of VAP21 expression even under the induced culture conditions. In such VAP21‐overexpressing cells, production of the vesicular stomatitis virus (VSV) was increased several‐fold, while suppression of the VAP21 expression resulted in reducing the VSV yields. From these results, we conclude that the VAP21 is a physiologically active cell membrane component of some animal species including the Syrian hamster, and might positively be involved in the VSV replication.
Prevalence of Infection and Genotypes of GBV‐C/HGV among Homosexual Men
Authors: Hattori Junko , Ibe Shiro , Nagai Hiromi , Wada Kaoru , Morishita Takayuki , Sato Katsuhiko , Utsumi Makoto , Kaneda Tsuguhiro 
Published: November 2013

AbstractSince the discovery of GB virus‐C (GBV‐C) and hepatitis G virus (HGV), many studies have been performed. These viruses are now known to be parenterally, as well as sexually transmitted. A phylogenetic analysis also revealed that GBV‐C has five major genotypes: type 1 predominates in West Africa, type 2 in Europe and the United States, type 3 in parts of Asia, type 4 in Southeast Asia, and type 5 in South Africa. Despite the number of reports so far, there have been few large‐scale surveys of homosexual men to determine the prevalence of the GBV‐C/HGV infections. We examined the levels of GBV‐C/HGV viremia in 297 homosexual men who attended the Nagoya Lesbian and Gay Revolution held in Nagoya, Japan. Reverse transcription‐polymerase chain reaction (RT‐PCR)/nested PCR of the GBV‐C/HGV 5′‐non‐coding region (NCR), and base sequence analyses showed that the infection rate was 12.5%, and genotypes in this population were classified into type 2 (32%) and type 3 (68%). None were classified as types 1, 4, or 5 in this study. Our results indicate that the GBV‐C/HGV type 2 seen mainly in Europe and the US is spreading widely in Japan, especially in the Nagoya district.
Ability of CD8+ T Cell Anti‐Feline Immunodeficiency Virus Activity Correlated with Peripheral CD4+ T Cell Counts and Plasma Viremia
Authors: Hohdatsu Tsutomu , Yamazaki Ayanori , Yamada Mitsunobu , Kusuhara Hajime , Kaneshima Takashi , Koyama Hiroyuki 
Published: November 2013

AbstractIn the host defense mechanism against feline immunodeficiency virus (FIV) infection, CD8+ T cells specifically attack virus‐infected cells and suppress the replication of the virus in a non‐cytolytic manner by secreting soluble factors. In this study, we measured CD8+ T cell anti‐FIV activity in 30 FIV‐infected cats. We investigated its relationship with the number of peripheral blood lymphocytes, particularly the CD4+ T cell and CD8+ T cell counts, and the relationship between anti‐FIV activity and the number of T cells of CD8α+βlo and CD8α+β− phenotypes. A clearly significant correlation was observed between anti‐FIV activity and the number of CD4+ T cells. A weaker anti‐FIV activity was associated with a greater decrease in the number of CD4+ T cells. However, there was no significant correlation between anti‐FIV activity and the number of B or CD8+ T cells. Compared with SPF cats, FIV‐infected cats had significantly higher CD8α+βlo T cell and CD8α+β− T cell counts, but, no significant correlation was observed between these cell counts and anti‐FIV activity. This anti‐FIV activity significantly correlated with plasma viremia, which was detected in cats with a weak anti‐FIV activity. These results suggest that the anti‐FIV activity of CD8+ T cells plays an important role in plasma viremia and the maintenance of CD4+ T cells in the body. It is unlikely that CD8α+βlo or CD8α+β− T cells appearing after FIV infection represent a phenotype of CD8+ cells with anti‐FIV activity.