bluuebunny/crossref_metadata_2025_split huggingface dataset:
Consensus Sequence on the Genes Encoding the Major Outer Surface Proteins (OspA and OspB) of Borrelia garinii Isolate
Authors: Jianhui Wang , Masuzawa Toshiyuki , Komikado Tetsuro , Yanagihara Yasutake 
Published: November 2013

AbstractJapanese Lyme borrelias classified as ribotype IV is predominant among isolates derived from clinical specimens, reservoir rodents and Ixodes persulcatus ticks, and has been characterized as Borrelia garinii. These B. garinii isolates have antigenic and genetic features apparently different from North American, European and other Asian isolates, especially in major outer surface proteins A (OspA) and B (OspB). In this study, we cloned and sequenced the genes encoding OspA and OspB from B. garinii strain FujiP2 (ribotype IV strain) isolated from I. persulcatus in Shizuoka, Japan. A sequence analysis revealed significant differences to the previously published sequences of ospA and ospB of B. burgdorferi sensu lato. The open reading frames of ospA and ospB consist of 822 and 888 nucleotides corresponding to the proteins of 273 and 295 amino acids, with molecular weights of 29,643 and 31,786 daltons, respectively. The most interesting finding is that the two osp genes share a consensus 282 bp sequence in their carboxy‐terminal portions and that the ospB gene is flanked by a 282 bp‐long direct repeat sequence. The deduced amino‐acid (aa) sequences of OspA and OspB of strain FujiP2 showed 60.1% homology, and have overall similarities of 70.5%, 70.3% and 75.6% to OspAB proteins of B. burgdorferi sensu stricto strain B31, Borrelia afzelii strain ACA1 and Borrelia garinii strain Ip90, respectively.
Characteristics and Pathogenicity of Non‐Melibiose‐Fermenting Strains of Yersinia pseudotuberculosis O3
Authors: Nagano Tetsuji , Ichimura Kazuhisa , Haji Norihiko , Nagao Kazuya , Someya Kazuhiko , Kiyohara Tomoko , Suzuki Kiyomi , Tsubokura Misao , Otsuki Koichi 
Published: November 2013

AbstractThe biological properties of non‐melibiose‐fermenting (NMF) strains of Yersinia pseudotuberculosis 03 were investigated. These strains were clearly distinguished from representative melibiose‐fermenting (MF) strains of Y. pseudotuberculosis 03 by their pathogenicity in mice, sensitivity to some phages, production of catalase, restriction endonuclease analysis of virulence plasmid DNA with BamHI, detection of specific yersinia outer‐membrane proteins with SDS‐PAGE, antigenicity of the outer‐membrane proteins and neutrophil resistance to phagocytosis. The pathogenicity of NMF strains was clearly less than that of MF strains. In addition, the resistance of NMF strains to phagocytosis and catalase activity was evidently weaker than that of MF strains. These results suggested that the difference of pathogenicity was due to the ability of catalase production. Although the relationship between the above characteristics and melibiose‐fermentation was not analysed, the pathogenicity of Y. pseudotuberculosis 03 strains can probably be predicted by testing melibiose‐fermentation and catalase production.
Production of a T‐Cell Clone Which Reacts with Membrane Proteins of Acholeplasma laidlawii
Authors: Kuwano Koichi , Ono Saori , Akashi Akira , Ohishi Masahiro , Shigematsu Hideki , Arai Sumio 
Published: November 2013

AbstractThe role of cellular immunity in mycoplasma infection is not completely understood. In this study, we established mycoplasma‐specific T‐cell clones to evaluate cellular immunity in mycoplasma infection. We developed a T‐cell clone (G‐10) which was stimulated with Acholeplasma laidlawii. The T‐cell clone G‐10, CD4+ and T‐cell receptor (TCR) αβ+ recognized the 42‐ and 65‐kilodalton (kDa) membrane proteins of A. laidlawii and responded to A. hippikon. Hence, the application of mycoplasma‐specific T cells such as G‐10 in animal models may allow the assessment of cellular immune response to mycoplasma infection.
Normal Human Fibroblasts Immortalized by Introduction of Human Papillomavirus Type 16 (HPV‐16) E6‐E7 Genes
Authors: Shiga Takashi , Shirasawa Hiroshi , Shimizu Kumiko , Dezawa Mari , Masuda Yoshiaki , Simizu Bunsiti 
Published: November 2013

AbstractThis report demonstrates that normal human fibroblasts can be immortalized by the introduction of HPV‐16 E6‐E7 genes. We designed zinc‐inducible expression plasmids with HPV‐16 E6, E7 or both. Each plasmid was introduced into normal human fibroblasts (TIG‐3 cells) using lipofection methods. Only trans‐fectants with the HPV‐16 E6‐E7 zinc‐inducible expression plasmid, which were cultured in medium supplemented with 100 μM ZnSO4, overcame crisis and could be cultured over 200 population doubling levels (PDLs). These cell lines showed the reactivation of telomerase after crisis, and morphological alterations were also observed.
Identification of Murine T Cells Reactive with the Bacterial Superantigen Yersinia pseudotuberculosis‐Derived Mitogen (YPM) and Factors Involved in YPM‐Induced Toxicity in Mice
Authors: Miyoshi‐Akiyama Tohru , Fujimaki Wakae , Yan Xao‐Jie , Yagi Junji , Imanishi Ken'ichi , Kato Hidehito , Tomonari Kyuhei , Uchiyama Takehiko 
Published: November 2013

AbstractWe previously reported that Yersinia pseudotuberculosis‐derived mitogen (YPM) acts as a superantigen to human T cells. In this study, we assessed the superantigenicity and toxicity of YPM using murine experimental models. YPM activated T cells to produce interleukin‐2 in a major histocompatibility complex class II molecule‐dependent manner. The T‐cell blasts induced by YPM expressed T‐cell receptor (TCR) β‐chain variable region (Vβ)7, VβS.1, Vβ8.2 and Vβ8.3. The injection of YPM into mice pre‐sensitized with D‐galactosamine induced lethal shock. This shock was blocked by the injection of monoclonal antibodies (mAbs) to CD4, TCR Vβ7 plus Vβ8, tumor necrosis factor‐α (TNF‐α) and interferon‐γ (IFN‐γ), but not by injection to CD8 or unrelated Vβ. These results indicate that YPM‐induced shock requires the presence of CD4+ T cells bearing TCR Vβ7 and Vβ8, and that endogenous TNF‐a and IFN‐γ mediate the lethal effects.
An ATP Bioluminescence Assay Applicable to Rapid Fluconazole Susceptibility Testing of Dermatophytes
Authors: Yoshida Tatsumi , Uchida Katsuhisa , Yamaguchi Hideyo 
Published: November 2013

AbstractAn ATP bioluminescence assay as a rapid reference method for fluconazole (FLCZ) susceptibility testing of dermatophytes, as well as yeasts, was developed and evaluated by comparing it with viability, turbidity and fungal protein content‐based conventional methods. FLCZ susceptibility results obtained with strains of Candida albicans and dermatophytes by the bioluminescence method in high‐resolution medium were well correlated with those obtained by conventional methods currently used in clinical microbiology laboratories or reported previously, including a broth dilution method by the National Committee for Clinical Laboratory Standards (NCCLS). Thus, ATP bioluminescence assay can be used to monitor fungal growth in liquid culture media. The procedure has considerable potential for the rapid testing of FLCZ susceptibility of dermatophytes and other fungi.
Genetic Control of Immune Responses to HIV‐1 env DNA Vaccine
Authors: Ishii Norihisa , Sugita Yasuyuki , Nakajima Hiroshi , Bukawa Hiroki , Asakura Yusuke , Okuda Kenji 
Published: November 2013

AbstractWe investigated the genetic control of immunoglobulin production and the delayed‐type hypersensitivity (DTH) response produced by an HIV‐specific DNA vaccine using several strains of mice. Murine antigen‐specific immunoglobulin production was determined by ELISA. The DTH response was assessed in terms of the footpad swelling reaction. All strains of mice, except for B10.RIII and B10.T(6R), exhibited strong immunoglobulin production and footpad swelling in response to the DNA vaccine. In vitro treatment of lymphoid cells with monoclonal antibodies showed that the footpad swelling response was mediated by CD4+8− and Ia— T cells. However, CD8+ T cells did not suppress footpad swelling. There was no difference in the induction of HIV‐specific immunoglobulin production or DTH response induced by the DNA vaccine among the strains, suggesting that HIV‐specific DNA vaccine is useful for immunizing various populations against HIV‐1.
Levels of Endogenous lnterleukin‐1, Interleukin‐6, and Tumor Necrosis Factor in Congenic Mice Infected with Borrelia garinii
Authors: Isogai Hiroshi , Kimura Koichi , Hayashi Shunji , Kubota Toru , Fujii Nobuhiro , Nishikawa Takeshi , Kotake Satoshi , Isogai Emiko 
Published: November 2013

AbstractThis study describes the levels of interleukin‐1 alpha (IL‐1α), tumor necrosis factor alpha (TNFα) and interleukin‐6 (IL‐6) in the sera and parenchymal organs of various congenic mouse strains infected with Borrelia garinii. A significant elevation of inflammatory cytokine levels was found in the organs of C3H/HeN (H‐2k) and B10.BR (H‐2k) mice but not in those of BALB/c mice (H‐2d). Focally produced cytokines can contribute to antimicrobial defense against these organisms. High levels of IL‐1α were observed in the sera of C3H/HeN, B10.BR and B10 (H‐2b) mice infected with B. garinii and they were associated with the presence of spirochetes in the skin. Thus, susceptible mice demonstrated a stronger cytokine response than resistant mice. This study presents in vivo evidence that B. garinii infection affects the immunopathogenesis of Lyme disease.
Replication of Feline Syncytial Virus in Feline T‐Lymphoblastoid Cells and Induction of Apoptosis in the Cells
Authors: Ikeda Yasuhiro , Itagaki Shin‐lchi , Tsutsui Shigeki , Inoshima Yasuo , Fukasawa Masashi , Tomonaga Keizo , Tohya Yukinobu , Maeda Ken , Mochizuki Masami , Miyazawa Takayuki , Mikami Takeshi 
Published: November 2013

AbstractFeline syncytial virus (FSV) was isolated from feline peripheral blood mononuclear cells of FSV‐seropositive cats. When the susceptibility of feline T‐lymphocytes to FSV was examined using three strains of FSV, FSV antigens were detected in the FSV‐infected T‐lymphoblastoid cells. Further, a diversity of biological properties, including replication kinetics and syncytia formation, was noted among the strains, and condensation of chromatin and the fragmentation of cellular DNA were observed in the infected cells. From these data, we conclude that FSV is lymphotropic and can induce apoptosis in the lymphocytes.
RobA‐lnduced Multiple Antibiotic Resistance Largely Depends on the Activation of the AcrAB Efflux
Authors: Tanaka Toshihiko , Horii Toshinobu , Shibayama Keigo , Sato Kumiko , Ohsuka Shinji , Arakawa Yoshichika , Yamaki Ken‐ichi , Takagi Kenzo , Ohta Michio 
Published: November 2013

AbstractRobA is a member of the XylS/AraC subfamily of DNA binding proteins, and when overexpressed, it induces multiple antibiotic resistance in Escherichia coli. In this study, we introduced a multicopy robA plasmid (pMEP1) and its derivative into OmpF mutants and an AcrAB‐deficient mutant. We found that a decrease in susceptibility to multiple antibiotics in these OmpF mutants when pMEP1 was introduced did not depend on OmpF porin expression. Interestingly, a ΔompF mutant (TK007) became more sensitive when pMEP1 was introduced. Moreover, no effect of RobA on the induction of multiple antibiotic resistance in an acrA1− mutant was observed. Therefore, we conclude that the multiple antibiotic resistance induced by the overexpression of RobA largely depends on the activation of the AcrAB efflux, as well as the activation of micF.
Successful Development of Air‐Dried Microplates (HP‐Plates) for Susceptibility Testing against Helicobacter pylori Isolates
Authors: Kawakami Yoshiyuki , Akahane Takayuki , Gotoh Akira , Okimura Yukie , Oana Kozue , Katsuyama Tsutomu 
Published: November 2013

AbstractWe have successfully developed and evaluated a new susceptibility testing procedure against Helicobacter pylori strains using air‐dried microplates “HP‐Plates” containing eight serially‐diluted anti‐H. pylori agents. HP‐Plate wells were reconstituted by the inoculation of 100 μl of H. pylori cell suspensions. After incubation at 37 C for 48 hr under humidified microaerophilic conditions, HP‐Plates were read visually with a circular mirror. We investigated the within‐day reproducibility tests of HP‐Plates using the six quality control (QC) strains we proposed. Of the 20 testings, determining the minimum inhibitory concentrations (MICs) of all the QC strains fell within ± 1 log2 dilution ranges. When 200 clinical isolates were tested with HP‐Plates and compared with the results obtained with the modified broth macrodilution method of NCCLS, more than 90% of the MICs also fell within ±1 log2 dilution ranges. We concluded that the HP‐Plate susceptibility test method is a practical and easily applicable alternative of susceptibility testing for clinical microbiology laboratories in determining the MICs of H. pylori isolates.
Triazine Dyes Inhibit HIV‐1 Entry by Binding to Envelope Glycoproteins
Authors: Hattori Toshio , Zhang Xiaoyan , Weiss Carol , Xu Younong , Kubo Toru , Sato Yoshimi , Nishikawa Shigemichi , Sakaida Hitoshi , Uchiyama Takashi 
Published: November 2013

AbstractWe have attempted to purify envelope (Env) glycoproteins of human immunodeficiency virus (HIV) from the culture supernatants of CHO‐Sec cells that secreted truncated 140‐kDa precursor and mature 120‐kDa Env glycoproteins. The concentrated culture supernatants were applied to a column coupled with cibacron blue 3GA (CB3GA) to separate albumin from the Env proteins because CB3GA, a triazine dye, has been known to have a high affinity to albumin. Unexpectedly, Env proteins as well as albumin bound to the column, and the bound Env proteins were eluted by increasing the ionic strength using KC1. Gp120 was eluted at 0.5–0.9 m of KC1, while a higher concentration (0.9–1.5 m) was necessary for the elution of gp140. The agarose gel coupled with reactive red 120 (RR120), another triazine dye with similar characteristics, also retained both Env proteins, and the bound Env proteins could be eluted in a similar manner. In addition, these agents inhibited syncytium formation caused by HTLV‐IIIB and HTLV‐IIMN. Inhibition was also seen when a virus‐free fusion assay between Env protein expressed in CHO cells and fluorescent labeled SupT1 cells were used. These findings indicate that triazine dyes bind to the functional regions of Env proteins of HIV‐1 that play important role(s) for HIV infection.
Cloning and Sequencing of the Vibrio parahaemolyticus fur Gene
Authors: Yamamoto Shigeo , Funahashi Tatsuya , Ikai Hisato , Shinoda Sumio 
Published: November 2013

AbstractA ferric uptake regulatory gene (fur) was cloned from Vibrio parahaemolyticus WP1 by a polymerase chain reaction‐based technique followed by functional complementation of a fur mutation in Escherichia coli. A sequence analysis showed that, at the amino acid level, the V. parahaemolyticus Fur protein is 81% identical with the Fur protein from E. coli and over 90% identical with those of the Vibrio species.
Morphological Features of a Filamentous Phage of Vibrio cholerae O139 Bengal
Authors: Shimodori Shoichi , Lida Kenichiro , Kojima Fumiko , Takade Akemi , Ehara Masahiko , Amako Kazunobu 
Published: November 2013

AbstractA filamentous phage was isolated from carrier strain AI‐1841 of Vibrio cholerae O139 Bengal and thus was termed fs phage. The phage was measured to be approximately 1 μm in length and 6 nm in width. One end of the phage was slightly tapered and had a fibrous appendage. The plaques developed on strain AI‐4450 of V. cholerae O139 were small and turbid. The phage grew in strain AI‐4450 and reached a size of 108 to 109 pfu/ml at 5 hr after infection without inducing any lysis of the host bacteria. The group of phages attached on rod‐shaped materials like fimbriae of this bacteria, with their fibrous appendages at the pointed end, were often found in the phage‐infected culture. The anti‐fimbrial serum effectively inhibited the infection of fs phage to the host strain AI‐4450. We thus concluded that the phage can be adsorbed on fimbriae with a fibrous appendage on the pointed end of the phage filament.
Interaction of Drugs for Eradication Therapy against Antibiotic‐Resistant Strains of Helicobacter pylori
Authors: Gotoh Akira , Kawakami Yoshiyuki , Akamatsu Taiji , Katsuyama Tsutomu 
Published: November 2013

AbstractTo clarify the interactions of drugs for combination therapy of Helicobacter pylori infection, especially due to antibiotic‐resistant strains, we have evaluated the in vitro effect of combining different drugs. Using a modified time‐kill assay, we tested the effect of combining 2 drugs from 4 agents; amoxicillin (AMPC), clarithromycin (CAM), metronidazole (MTZ) and lansoprazole (a proton pump inhibitor). The H. pylori in the study consisted of 4 strains sensitive to the all drugs, 2 strains resistant only to CAM, 2 strains resistant only to MTZ, and 2 strains resistant to both CAM and MTZ. From the 6 different drug combinations, synergism was observed for 5 of the combinations, among which the combination of AMPC and CAM revealed such effects most frequently. However, all of the strains which showed synergism were sensitive to both of the drugs. In the case of the strains resistant to CAM and/or MTZ, no synergism was demonstrated in any of the combinations including CAM and/or MTZ. When a strain was resistant to one drug from a combination, no synergism was detected. Thus, the administration of a drug to which the strains are resistant may have no advantage in the eradication therapy of H. pylori. For a more effective and safer therapy, susceptibility testing should be performed before treatment.
Detection of EBV RNA (EBER‐1 and EBER‐2) in Malaria Lymph Nodes by In Situ Hybridization
Authors: Facer Christine , Khan George 
Published: November 2013

AbstractAcute Plasmodium falciparum malaria in African children allows expansion of latent Epstein‐Barr virus infection, leading to colonization of lymph nodes by virus‐infected lymphoblasts in 60% of cases as demonstrated by in situ hybridization for the detection of EBER‐1 and EBER‐2 RNA. This probably arises against a background of malaria‐induced immunosuppression to EBV and concurrent lymphoid activation. The relevance of the results to the pathogenesis of African endemic Burkitt's lymphoma is discussed.
Shotgun Cloning and Characterization of the Thymidylate Synthase‐Encoding Gene from Mycobacterium bovis BCG
Authors: Matsumoto Sohkichi , Yukitake Hideharu , Ohara Naoya , Dairi Tohru , Kanbara Hiroji , Yamada Takeshi 
Published: November 2013

AbstractThe shotgun cloning of a Mycobacterium bovis BCG (BCG) genome into pBluescript SK (+) successfully yielded a 0.9 kbp fragment, confirming the ability of Escherichia coli thyA mutant MH2702 to grow in a thymine‐depleted medium. This DNA fragment contained a gene homologous to the thymidylate synthase (TS)‐encoding genes (thyA) of other organisms. An inverted repeat sequence and open reading frame (ORF) were observed at the upstream region of the thyA. A computer analysis revealed that the protein encoded by this ORF possessed a structure unique for a DNA binding protein.
Biochemical and Immunological Analyses of the Flagellin of Clostridium tyrobutyricum ATCC 25755
Authors: Arnold Françoise , Bédouet Laurent , Batina Pierre , Robreau Georges , Talbot Françoise , Lécher Pierre , Malcoste Roger 
Published: November 2013

AbstractThe monoclonal antibody 21E7‐B12 (IgG3) can be used in a direct method of Clostridium tyrobutyricum detection based on an immunoenzymatic assay. Immunoelectron microscopy demonstrated that the 21E7‐B12 antibody recognized the surface‐exposed epitopes on the flagellar filaments of C. tyrobutyricum. After flagellar extraction, the purified flagellin showed an apparent molecular mass of 46 kDa with an isoelectric point of 3.6. Sugar staining, mild periodate oxidation and é‐elimination experiments showed that the flagellin was glycosylated and that the 21E7‐B12 epitope was located in the sugar moiety. Amino acid composition showed that the flagellar filament protein contained a high percentage of serine and threonine, while proline was absent. The first 23 residues of the N‐terminal were determined and sequence homology with other flagellins was found.
Follow‐Up Study of Hypervariable Region Sequences of the Hepatitis C Virus (HCV) Genome in an Infant with Delayed Anti‐HCV Antibody Responses
Authors: Katayama Yuko , Tajiri Hitoshi , Tada Kanae , Okada Shintaro , Tong Wen‐yan , Ishido Satoshi , Hotta Hak 
Published: November 2013

AbstractAn infant born prematurely and infected with hepatitis C virus (HCV) one month after birth was followed for 4.5 years. The patient did not produce detectable anti‐HCV antibodies until two years after the onset of hepatitis. Before seroconversion, a single clone of HCV, as determined by quasispecies of the hypervariable region (HVR) of the HCV genome, was almost exclusively found in the serum. After seroconversion, however, another distinct lineage of HCV clones replaced it within half a year. As HCV infection persisted further in the presence of anti‐HCV antibodies, many derivatives of both sequence lineages emerged to exhibit the typical quasispecies feature of HVR sequences. Neither seroconversion nor the changes in HVR sequences influenced the serum aminotransferase titers.
Partial Analysis of the Flagellar Antigenic Determinant Recognized by a Monoclonal Antibody to Clostridium tyrobutyricum
Authors: Bédouet Laurent , Arnold Francoise , Robreau Georges , Batina Pierre , Talbot Francoise , Malcoste Roger 
Published: November 2013

AbstractIn order to count Clostridium tyrobutyricum spores in milk after membrane filtration, murine 21E7‐B12 monoclonal antibody was produced. Elution of the monoclonal antibody from this antigen, the flagellar filament protein, by carbohydrate ligands was used to study the epitope structure. A competitive elution of an anti‐dextran monoclonal antibody by carbohydrate ligands served as a control in order to validate the immunological tool applied to flagellin epitope study. The carbohydrate moiety of flagellin contained D‐glucose and N‐acetyl‐glucosamine in a molar ratio of 11:1 as determined by gas‐liquid chromatography and 2 low‐abundancy unidentified compounds. In ELISA, D‐glucose and N‐acetyl‐glucosamine did not dissociate the antibody‐flagellin complex contrary to maltose, maltotriose, maltotetraose and maltopentaose. The efficiency of elution increased from the dimer to the pentamer and became nil for maltohexaose and maltoheptaose. The fact that the hexamer and heptamer could not react with the 21E7‐B12 monoclonal antibody could be explained by a drastic conformational change. The overall stretched maltopentaose switch to a helical‐shaped maltoheptaose which could not fit the 21E7‐B12 monoclonal antibody antigen‐combining site. Thus, flagellin epitope may contain α(1→4) linked glucose residues plus either N‐actyl‐glucosamine or an unidentified compound that maintain it in an extended shape.