bluuebunny/crossref_metadata_2025_split huggingface dataset:
Isolation of K88 Antigen Variants (ab, ac, ad) from Porcine Enterotoxigenic Escherichia coli Belonging to Different Serotypes
Authors: González Enrique A. , Vázquez Froilán , Garabal José Ignacio , Blanco Jorge 
Published: November 2013

AbstractFimbriae isolation by means of thermal shock was applied to fifteen K88‐positive (three K88ab, nine K88ac and three K88ad) Escherichia coli reference strains belonging to serotypes O8:K87, O32, O45, O138:K81, O141:K85, O147:K89, O149:K91, and O157, as well as to ten K88‐positive enterotoxigenic strains isolated from porcine diarrhea in Spain, all of them belonging to the O149 serogroup. Fimbriae were removed from the bacterial cells by thermal shock at 60 C and then precipitated using ammonium sulfate. The final amount of K88 antigen and the purification degree were not related to the serogroup of the bacteria or to the antigen variant but were related to the buffer used for isolation. Phosphate buffer containing urea was shown to be more effective than Tris‐HCl for isolation of K88 antigen. The molecular weights by SDS‐PAGE for K88ab, K88ac, and K88ad were 28.5, 29.2, and 31.0 kDa, respectively. All enterotoxigenic E. coli strains isolated in Spain showed the K88ac variant.
Evaluation of Antiherpetic Compounds Using a Gastric Cancer Cell Line: Pronounced Activity of BVDU against Herpes Simplex Virus Replication
Authors: Kodama Eiichi , Igarashi Akira , Mori Shuichi , Hashimoto Kou‐Ichi , Suzuki Toshimitsu , De Clercq Erik , Shigeta Shiro 
Published: November 2013

AbstractWe developed a rapid and simple method for the screening of antiviral agents against herpes simplex virus (HSV) in a model of gastrointestinal herpetic infection in vitro. This method was based on inhibition of HSV‐induced cytopathogenicity in gastric adenocarcinoma MKN‐28 cells, as monitored by an MTT colorimetric assay. From the various compounds that were evaluated for their activity against HSV‐1 and HSV‐2, brivudine (BVDU) emerged as the most effective. When the 50% effective concentration (EC50) values of the antiherpes agents in MKN‐28 cells were compared with those in human embryo lung MRC‐5 cells, all compounds, except for BVDU, showed higher EC50 values in MKN‐28 cells. For BVDU the EC50 values in MKN‐28 cells were 0.8 (HSV‐1) and 0.036 (HSV‐2) times the EC50 values in MRC‐5 cells. Thus BVDU was 27.5 times more active against HSV‐2 in MKN‐28 cells than in MRC‐5 cells. The MKN‐28 gastric cancer cells may be useful for the rapid screening of anti‐HSV agents and, in particular, those that may be useful in therapy of gastrointestinal HSV infections in gastrointestinal herpetic infection.
Stability of Hemagglutinating Activity of Extracellular and Intracellular Forms of Japanese Encephalitis Virus Exposed to Acidic pH
Authors: Nawa Masaru 
Published: November 2013

AbstractThe hemagglutinating (HA) activity of extracellular and intracellular forms of Japanese encephalitis (JE) virus was comparatively titrated by exposure to acidic pH below 7.0. A pH‐dependent irreversible loss in titer was observed with the virus grown in both C6/36 and BHK 21 (BHK) cells maintained in the pH range of 5.8 to 7.0 for 10 min at 37 C. The HA activity of intracellular virus was relatively more stable than that of extracellular virus in the pH range of 5.8 to 6.4. Virion structural components, envelope glycoprotein (E), capsid (C), and membrane (M) proteins in extracellular virus and E, C, and the precursor form of M (prM) proteins in intracellular virus were detected by SDS‐polyacrylamide gel electrophoresis (SDS‐PAGE) and immunoblotting. A panel of monoclonal antibody (mAb) directed for nine antigenic epitopes on the JE virus E protein molecule was used for the analysis of antigenic reactivity of E protein after treatment at pH 6.0. The reaction between the extracellular virus and three HA‐inhibiting (HI) mAbs was significantly reduced after acid treatment; however, the antigenic reactivity of intracellular virus was much more stable with a 100‐ to 1,000‐fold difference. Infectivity titers of extracellular and intracellular viruses in Vero cells were reduced by 1/24,100 and 1/21,666 after acidic treatment at pH 6.0. In contrast, the infectivity of intracellular viruses was more stable, with residual infectivity of 1/182 and 1/340 for BHK and C6/36 cell‐grown virus, respectively. Acidic treatment of JE virus not only resulted in the irreversible loss of its HA activity but also affected the antigenic reactivity of HI epitopes on its E protein molecule.
In Vitro Effects of Immunosuppressive Agents on Cytokine Production by HTLV‐I‐Infected T Cell Clones Derived from the Ocular Fluid of Patients with HTLV‐I Uveitis
Authors: Sagawa Kimitaka , Mochizuki Manabu , Katagiri Kazuko , Tsuboi Izumi , Sugita Sunao , Mukaida Naofumi , Itoh Kyogo 
Published: November 2013

AbstractThe present study was designed to investigate the in vitro effects of potential therapeutic agents on cytokine production by five HTVL‐I‐infected T cell clones (TCC) established from the ocular fluid of patients with HTLV‐I uveitis. Each of the five HTLV‐I‐infected TCC was cultured at 1 × 106 cells/ml with or without an immunosuppressive agent (hydrocortisone, FK506, rapamycin, indomethacin, or prostaglandin E2) for 22 hr in humidified 5% CO2 in air at 37 C. The production of various cytokines in the culture supernatant from each TCC was measured by ELISA. The HTLV‐I‐infected TCC produced high amounts of IL‐1α, IL‐3, IL‐6, IL‐8, TNF‐α, IFN‐γ, and GM‐CSF, and low but significant levels of IL‐2 and IL‐10 without any stimuli. Hydrocortisone severely depressed the production by these TCC of all the cytokines except for IL‐2, which was slightly increased. Prostaglandin E2 depressed the production of IL‐1α, while it up‐regulated the production of IL‐6, TNF‐α, and IFN‐γ. Rapamycin depressed the production of IL‐6 and TNF‐α, and FK506 depressed the production of TNF‐α. Hydrocortisone also severely depressed the cytokine production by PHA‐stimulated peripheral blood mononuclear cells obtained from healthy volunteers. Of the immunosuppressive agents tested, hydrocortisone exhibited the strongest suppression of cytokine production by HTLV‐I‐infected TCC. This result was in agreement with the in vivo effects of hydrocortisone in patients with HTLV‐I uveitis. These TCC will be useful in investigating the effects of potential therapeutic agents for HTLV‐I uveitis in vitro.
Differential Regulation of HLA‐DR Expression and Antigen Presentation in Toxoplasma gondii‐Infected Melanoma Cells by Interleukin 6 and Interferon γ
Authors: Yang Tian‐Hui , Aosai Fumie , Norose Kazumi , Ueda Masakatsu , Yano Akihiko 
Published: November 2013

AbstractCD4+ cytotoxic T lymphocytes (CTL) clones, YT‐4 and YT‐9, specific for Toxoplasma gondii (T. gondii)‐infected melanoma SK‐MEL 28 (P36), were generated from the peripheral blood lymphocytes (PBL) of a patient with chronic toxoplasmosis. These CTL clones were shown to secrete significant amounts of interleukin 6 (IL‐6) and interferon γ (IFN‐γ) upon antigen (Ag)‐specific stimulation. Downregulation of human leukocyte antigen (HLA)‐DR surface expression and HLA‐DR mRNA levels in P36 cells were observed when P36 cells were infected with T. gondii. Such downregulated HLA‐DR expressions of 71 gondii‐infected P36 cells were upregulated by treatment with both recombinant IL‐6 (rIL‐6) and recombinant IFN‐γ (rIFN‐γ). The antigen‐presenting ability of T. gondii‐infected P36 cells to T. gondii‐infected cell‐specific CTL was enhanced by rIFN‐γ but not by rIL‐6. The present study reveals the existence of differential regulation of HLA‐DR expression and Ag presentation in T. gondii‐infected melanoma cells by IL‐6 and IFN‐γ.
Hepatitis B Virus with X Gene Mutation Is Associated with the Majority of Serologically “Silent” Non‐B, Non‐C Chronic Hepatitis
Authors: Fukuda Ryo , Ishimura Norihisa , Kushiyama Yoshinori , Moriyama Nobuyuki , Ishihara Shunji , Chowdhury Aktaruzzaman , Tokuda Alejandro , Sakai Shino , Akagi Shuji , Watanabe Makoto , Fukumoto Shiro 
Published: November 2013

AbstractHepatitis B virus (HBV) with X gene mutations has been a putative pathogen of chronic hepatitis without serological markers of known hepatitis viruses. The aim of this study was to reconfirm whether the HBV with the X gene mutation is associated with these serologically “silent” non‐B, non‐C (NBNC) chronic hepatitis, alcoholic liver disease (ALD) and autoimmune hepatitis (AIH). HBV DNA was amplified from serum and sequenced in 30 patients with NBNC chronic hepatitis in comparison with 20 patients with ALD and 5 patients with AIH. HBV DNA was identified in 21 patients (70%) in NBNC chronic hepatitis by nested polymerase chain reaction while only one patient (5%) in ALD and none in AIH showed HBV DNA. Eighteen (85.7%) of the 21 identified HBV DNAs had an identical 8‐nucleotide deletion mutation at the distal part of the X region. This mutation affected the core promoter and the enhancer II sequence of HBV DNA and created a translational stop codon which truncated the X protein by 20 amino acids from the C‐terminal end. All the HBV DNAs had a precore mutation at the 83rd nucleotide resulting in disruption of HBe antigen synthesis. These results indicate that HBV mutants are closely associated with the majority of serologically “silent” NBNC chronic hepatitis cases and the population of such mutant HBV DNAs is not uniform.
Quantification of the CD55 and CD59, Membrane Inhibitors of Complement on HIV‐1 Particles as a Function of Complement‐Mediated Virolysis
Authors: Nakamura Mariko , Okada Hidechika , Sasaki Hiroyuki , Yoshida Kiyotsugu , Kamada Minori , Okada Noriko , Terada Masaki , Ohno Tsuneya 
Published: November 2013

AbstractPrevious studies have demonstrated that the murine monoclonal antibody (MoAb) NM‐01 activates the human complement classical pathway resulting in lysis of human immunodeficiency virus (HIV). The present study was performed to determine the availability of the V3‐loop of gp120 relative to the complement regulatory proteins, CD55 (DAF) and CD59 (HRF20) molecules on HIV. The results demonstrate that CD55 and CD59 exist on HIV virions, along with gp120 molecules. These findings suggest that activation of human complement on free viral particles is induced by MoAb NM‐01 and that this occurs regardless of the presence of CD55 and CD59 molecules. The destruction of viral particles was demonstrated by a decrease in infectivity. The involvement of human complement in this process was confirmed with an immunoelectron microscopy technique by the presence of a human C9 to prove membrane attack complex (MAC). The results indicate that NM‐01 can induce complement activation because of the ratios of CD55 and CD59 to gp120 molecules on HIV virions. The availability of the gp120 V3 domain on the virion is sufficient for binding of NM‐01 and thereby the formation of MAC that results in virolysis.
Does Coinfection ofBartonella henselaeand FIV Induce Clinical Disorders in Cats?
Authors: Ueno Hiroshi , Hohdatsu Tsutomu , Muramatsu Yasukazu , Koyama Hiroyuki , Morita Chiharu 
Published: November 2013

AbstractIt was found thatBartonella henselae (B. henselae)may induce clinical disorders in cats in natural conditions from a comparison of the serological status forB. henselaewith the serostatus for feline immunodeficiency virus (FIV) and several clinical characteristics in 170 domestic cats. Seropositivity forB. henselaewas not significantly different between FIV antibody‐positive and ‐negative cats (18.4% vs 16.0%). The incidence of clinical characteristics were compared among four cat groups distinguished by the reactivity of sera againstB. henselaeand FIV. The incidence of lymph node swelling was lower in only FIV antibody‐positive cats (3.0%), but higher inB. henselaeantibody‐positive cats (13.6%) and significantly higher in bothB. henselaeand FIV antibody‐positive cats (42.9%) compared with the incidence of lymph node swelling in cats which were negative for both antibodies (5.5%). The same relation was also observed for the incidence of gingivitis among the 4 cat groups, suggesting that coinfection ofB. henselaeand FIV may be associated with gingivitis and lymphadenopathy in cats.
Lipopolysaccharides of Escherichia coli K12 Strains That Express Cloned Genes for the Ogawa and Inaba Antigens of Vibrio cholerae O1: Identification of O‐Antigenic Factors
Authors: Hisatsune Kazuhito , Kondo Seiichi , Iguchi Takehiro , Ito Teruyo , Hiramatsu Keiichi 
Published: November 2013

AbstractStructural and serological studies were performed with the lipopolysaccharide (LPS) expressed by Escherichia coli K12 strains No. 30 and No. 64, into which cosmid clones derived from Vibrio cholerae O1 NIH 41 (Ogawa) and NIH 35A3 (Inaba) had been introduced, respectively. The two recombinant strains, No. 30 (Ogawa) and No. 64 (Inaba), produced LPS that included, in common, the O‐polysaccharide chain composed of an α(1 → 2)‐linked N‐(3‐deoxy‐L‐glycero‐tetronyl)‐D‐perosamine (4‐amino‐4,6‐dideoxy‐D‐manno‐pyranose) homopolymer attached to the core oligosaccharide of the LPS of E. coli K12. Structural analysis revealed the presence of N‐(3‐deoxy‐L‐glycero‐tetronyl)‐2‐O‐methyl‐D‐perosamine at the non‐reducing terminus of the O‐polysaccharide chain of LPS from No. 30 (Ogawa) but not from No. 64 (Inaba). Serological analysis revealed that No. 30 (Ogawa) and No. 64 (Inaba) LPS were found to share the group antigen factor A of V. cholerae O1. They were distinguished by presence of the Ogawa antigen factor B [co‐existing with relatively small amounts of the Inaba antigen factor (c)] in the former LPS and the Inaba antigen factor C in the latter LPS. It appears, therefore, that No. 30 (Ogawa) and No. 64 (Inaba) have O‐antigenic structures that are fully consistent with the AB(c) structure for the Ogawa and the AC structure for the Inaba O‐forms of V. cholerae O1, respectively. Thus, the present study clearly confirmed our previous finding that the Ogawa antigenic factor B is substantially related to the 2‐O‐methyl group at the non‐reducing terminus of the α(1 → 2)‐linked N‐(3‐deoxy‐L‐glycero‐tetronyl)‐D‐perosamine homopolymer that forms the O‐polysaccharide chain of LPS of V. cholerae O1 (Ogawa).
Kinetics of Organ‐Associated Natural Killer Cells and Intermediate CD3 Cells during Pulmonary and Hepatic Granulomatous Inflammation Induced by Mycobacterial Cord Factor
Authors: Tabata Akihiro , Kaneda Kenji , Watanabe Hisami , Abo Toru , Yano Ikuya 
Published: November 2013

AbstractWe investigated here the kinetics of natural killer (NK) cells and extrathymic T cells, which include intermediate CD3 cells and γδ T cells, in the cord factor‐induced granulomatous inflammation of the lungs and liver. In Balb/c mice, pulmonary inflammation elevated the proportion of NK cells and that of extrathymic T cells to mononuclear cells in the lungs. C3H/He mice exhibited shorter‐term inflammation of the lungs than Balb/c mice and accordingly showed a smaller increase in the proportions of pulmonary NK cells and intermediate CD3 cells. In the liver of Balb/c mice, hepatic NK cells increased as well with the granulomatous changes, while intermediate CD3 cells exhibited a transient decrease before they increased. The present study has demonstrated that granulomatous inflammation is accompanied by the increase of lung‐associated NK cells and extrathymic T cells and that there exists a difference between these two mouse strains in the induction of these lymphocyte subsets by cord factor.
Purified Protein from Salmonella typhimurium Inhibits the Interleukin‐2 Response of Murine Splenic T‐Lymphocytes Activated with Anti‐CD3 Antibody
Authors: Matsui Katsuhiko 
Published: November 2013

AbstractPreviously, we demonstrated that the immunosuppression induced by a purified preparation of Salmonella typhimurium‐derived inhibitor of T‐cell proliferation (STI) can be observed in terms of suppression of the proliferation of murine spleen cells stimulated with a mitogenic lectin. In the present study, I observed that STI inhibited the interleukin‐2 (IL‐2) response of purified murine splenic T lymphocytes stimulated with anti‐CD3 antibody. The flow cytometric analysis of IL‐2 receptor (IL‐2R) expression on T cells showed that STI specifically suppressed the expression of IL‐2Rβ and IL‐2Rγ. Furthermore, when the IL‐2‐dependent T‐cell line CTLL‐2 was incubated with STI, the growth of CTLL‐2 cells was significantly inhibited. These results suggest that the target cells for STI are T cells themselves, and that the suppression of T‐cell proliferation induced by STI might involve a defect in the IL‐2 receptor (IL‐2R) function of T cells.
Adhesiveness for Extracellular Matrices and Lysosomal Enzyme Release from Normal andβ2Integrin‐Deficient Bovine Neutrophils
Authors: Nagahata Hajime , Higuchi Hidetoshi , Noda Hiroshi , Tamoto Koichi , Kuwabara Mikinori 
Published: November 2013

AbstractThe adhesiveness of control and CD18‐deficient bovine neutrophils on culture plates precoated with collagen I, collagen IV, fibronectin and laminin was measured to evaluate the possible factors for adherence to extracellular matrices. The release ofN‐acetyl‐β‐D‐glucosaminidase (NAGase) from control and CD18‐deficient neutrophils stimulated with complement receptor type 3 (CR3) or Fc receptor dependent stimuli was also evaluated. The adhesive activities of CD18‐deficient neutrophils to collagen I, collagen IV and fibronectin were significantly diminished (P< 0.05); however, similar adhesion to laminin was observed in CD18‐deficient neutrophils and control neutrophils. The adhesive activity of control neutrophils on uncoated plates increased 2.5 times (P< 0.05) with the presence of PMA. The mean activities for NAGaserelease from CD18‐deficient neutrophils stimulated with opsonized zymosan and aggregated bovine immunoglobulin G (Agg‐IgG) were 46.7 and 82.7% that of the control neutrophils, respectively. The Agg‐IgG‐induced NAGaserelease from control and CD18‐deficient neutrophils was eliminated by H7, a protein kinase C inhibitor. These results support that an association between CR3 and Fc receptors on neutrophils appears to play an essential role in neutrophil functions.
Coxsackievirus B3‐Induced Chronic Myocarditis in Mouse: Use of Whole Blood Culture to Study the Activation of TNFα‐Producing Cells
Authors: Hober Didier , Andreoletti Laurent , Shen Lu , Copin Marie‐Christine , Desmidt Angelique , Wattré Pierre 
Published: November 2013

AbstractThe pathogenesis of CVB3‐induced chronic myocarditis remains unknown. Activated monocytes and macrophages may maintain ongoing inflammation during a persistent CVB3 infection and possibly represent the major mechanism leading to chronic myocarditis. We decided to study the activation status of cells by studying TNFα secretion in vitro using whole blood culture in CVB3‐induced murine chronic myocarditis. Seven DBA/2 +/+ mice and 18 NMRI nu/nu mice were inoculated intraperitoneally with 5 × 105 pfu of CVB3, and mice were mock‐infected. Thirty‐one days post‐infection, all mice were sacrificed, blood samples were obtained from the heart, and the heart was removed. Enteroviral genomic detection by RT‐PCR, virus isolation and histological analysis of heart samples were performed. Heparinized whole blood (25 μl) was cultured for 4 hr and 24 hr in sterile 96 well‐plate containing 225 μl RPMI in the presence or the absence of activators (LPS + PHA). The TNFα levels in the whole blood from mock‐infected DBA/2 (n = 4) and NMRI nu/nu mice (n = 5) were not different. A moderate increase of TNFα was observed in three out of five DBA/2 mice with negative CVB3 that had no histological abnormalities in myocardium. An increased level of TNFα was found in the sole DBA/2 mouse with positive CVB3 detection and chronic myocarditis. An increased level of TNFα was found in one out of nine NMRI nu/nu mice with positive CVB3 detection and chronic myocarditis and in one out of seven mice with positive CVB3 detection exempt of lesions in myocardium. In other infected mice, the level of TNFα was normal. Enteroviral genome was not detected in the blood from infected mice at 31 days post‐infection. The increased TNFα level in some mice may be designed for a beneficial inflammatory and immune response, however, an exaggerated release may be associated with an adverse effect. The normal TNFα level in whole blood cultures from mice with chronic myocarditis does not exclude enhanced cytokine production at infected loci such as myocardial tissue. This is the first report to use whole blood cultures to study the production of cytokines in virus‐induced disease in a small animal model.
Molecular Epidemiology of Japanese Encephalitis Virus in Okinawa
Authors: Ma Shao‐Ping , Arakaki Sakae , Makino Yoshihiro , Fukunaga Toshihiko 
Published: November 2013

AbstractIn order to elucidate the molecular characteristics of Japanese encephalitis (JE) virus in Okinawa, 23 strains of JE virus isolated in a 25‐year span were sequenced for the 240 nucleotides of the C‐preM junction region and 111 nucleotides of the E gene region and compared with those of reference strains isolated in mainland Japan. The results of phylogenic analysis showed that although all the Okinawan isolates showed more than 96% homology in the nucleotide sequence in each region, they were chronologically divided into two groups: the old group (nine strains) and a new group (14 strains). On the other hand, in a comparison with reference strains in mainland Japan, the Okinawan isolates showed more than 94% nucleotide sequence homology in both regions, indicating that the Okinawan strains belong to the same genotype as that of JE strains in mainland Japan. The nucleotide homology of the old group was relatively higher than that of the new group. Among the 14 strains in the new group, 13 strains were isolated from mosquitoes collected from a pig farm from 1986 through 1992. These strains showed higher nucleotide divergence than the old group strains, isolated from mosquitoes and swine sera collected at several sites, in both regions. A nucleotide substitution at the position 1920 in the E gene was identified in three isolates. This substitution generated an asparagine‐proline‐threonine sequence capable of serving as an attachment site of carbohydrate.
The UL2 Open Reading Frame of Bovine Herpesvirus 1 Encodes a Uracil‐DNA Glycosylase
Authors: Chung Yang‐Tsung , Hsu Weili 
Published: November 2013

AbstractSequence analysis within the unique long segment of the bovine herpesvirus 1 (BHV‐1) genome previously identified an open reading frame (ORF), designated UL2, whose deduced polypeptide of 204 amino acids contained a consensus uracil‐DNA glycosylase (UDGase) signature sequence. To determine whether the BHV‐1 UL2 ORF product has UDGase activity, we positioned the UL2 sequence downstream of the T7 promoter on the vector pET‐28b(+) and expressed it in Escherichia coli. Upon induction with isopropyl β‐D‐thiogalactopyranoside these cells produced a 23‐kDa protein, the molecular mass of which was in accordance with the prediction from the nucleotide sequence. A one‐step purification procedure using nickel‐chelating affinity chromatography resulted in a homogeneous preparation of this protein, which displayed specific UDGase activity in an in vitro enzyme assay. These results provide evidence that the BHV‐1 UL2 gene does encode a UDGase.
Proliferation and Teratogenicity of Aino Virus in Chick Embryos
Authors: Kitano Yoshio , Ohzono Hiroyuki , Shimizu Tsutomu 
Published: November 2013

AbstractAino virus (AIV; JaNAr 28 strain) 103 TCID50/0.2 ml was inoculated in the yolk sac of 8‐day‐old chick embryos. Recovery and titration of the virus from various organs including the central nervous system (CNS) and skeletal muscle were performed at 2, 4, 7, 10 and 13 days after inoculation (PI). AIV was systemically disseminated and proliferated even 2 days PI. The titers of the recovered virus from the CNS and from skeletal muscle was the highest at 4 days PI and declined with time, whereas hydranencephaly, arthrogryposis and cerebellar hypoplasia developed at 7 days PI and gradually progressed until 13 days PI.