bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pi30 DNA Probe May Be Useful for the Identification of Prevotella intermedia at the Species or Strain Level
Authors: Shin Yong Kook , Jeong Seung‐U , Yoo So Young , Kim Mi‐Kwang , Kim Hwa‐Sook , Kim Byung‐Ock , Kim Do Kyung , Hwang Ho‐Keel , Kook Joong‐Ki 
Published: November 2013

AbstractRecently, we introduced a new method for the rapid screening of bacterial species‐ or subspecies‐specific DNA probes, named the “inverted dot blot hybridization screening method.” This method has subsequently been then applied to develop species‐ or strain‐specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In a previous study, the inverted dot blot hybridization data showed that a probe, Pi30, was specific for P. intermedia. In this study, the DNA probe Pi30 was evaluated by Southern blot analysis to determine if it could distinguish P. intermedia from P. nigrescens. The data showed that the probe Pi30 reacted with the genomic DNAs from the reference strains and clinical isolates of both P. intermedia and P. nigrescens, but the size of the signal bands was different. In addition, the probe Pi30 reacted with a 1.4 kbp fragment from the genomic DNAs digested with PstI of the P. intermedia strains but not with any fragments of P. nigrescens strains. The result indicates that the probe Pi30 could be useful for the identification of P. intermedia by restriction fragment length polymorphism (RFLP) at the species or strain level.
Isolation of Mini‐Tn5Km2 Insertion Mutants of Salmonella enterica Serovar Oranienburg Sensitive to NaCl‐Induced Osmotic Stress
Authors: Asakura Hiroshi , Panutdaporn Nantika , Kawamoto Keiko , Igimi Shizunobu , Yamamoto Shigeki , Makino Sou‐ichi 
Published: November 2013

AbstractWe previously reported that the viability of Salmonella Oranienburg strains under NaCl stress was variable and depended on the strain's origin; food strains were resistant and patient strains sensitive to NaCl. Therefore, we mutagenized a food strain with a mini‐Tn5Km2 transposon. Of 2,400 mutants screened, 15 NaCl‐sensitive mutants were isolated, and 7 genes associated with NaCl‐sensitivity were identified. The intact genes complemented their own food‐strain mutants, but not patient‐strain mutants, suggesting that the difference in NaCl‐sensitivity between food and patient S. Oranienburg strains might not arise from a single gene mutation, but from change in multiple osmoregulatory mechanisms in Salmonella.
Detection of Tannerella forsythia and/or Prevotella intermedia Might Be Useful for Microbial Predictive Markers for the Outcome of Initial Periodontal Treatment in Koreans
Authors: Kook Joong‐Ki , Sakamoto Tomonori , Nishi Kazuya , Kim Mi‐Kwang , Seong Jin‐Hyo , Son Young Nam , Kim Dong‐Kie 
Published: November 2013

AbstractA proportion of diseased sites in periodontal disease do not respond to the initial treatment, which might be due in part to the presence of specific microbial pathogens. The aim of this study was to clarify the value of microbial screening for predicting the outcome of periodontal treatment in Koreans using a polymerase chain reaction (PCR). This study enrolled 32 adults with periodontal disease. Microbial and clinical examinations were performed at the baseline and after the initial treatment (professional toothbrushing, scaling, and root planing). Subgingival plaque samples were taken from four sites in each subject (total 128 samples). PCR was used to detect the four putative pathogenic bacteria. There was an improvement in the average of each clinical measurement after the initial treatment. However, approximately half of the sites exhibiting bleeding upon probing (BOP) at the baseline still exhibited bleeding after treatment. There was a close association between the presence of BOP and the presence of Tannerella forsythia (formerly Bacteroides forsythus) and/or Prevotella intermedia. Furthermore, the sites harboring both T. forsythia and P. intermedia at the baseline had a poorer response to treatment than the sites where these two species were not detected. Therefore, microbial screening for T. forsythia and P. intermedia might be useful for predicting the treatment outcome in Koreans.
Induction of Inducible Nitric Oxide (NO) Synthase mRNA and NO Production in Macrophages Infected with Influenza A/PR/8 Virus and Stimulated with Its Ether‐Split Product
Authors: Imanishi Nobuko , Andoh Tsugunobu , Sakai Shinya , Satoh Miyuki , Katada Yuko , Ueda Kyouka , Terasawa Katsutoshi , Ochiai Hiroshi 
Published: November 2013

AbstractWe investigated the inductive activity of infective influenza A/PR/8/34 (PR8) virus and its ether‐split product (ESP) on the expression of inducible nitric oxide (NO) synthase (iNOS) and NO production in RAW264.7 (RAW) cells, a murine macrophage (MΦ) cell line, and thioglycolate‐elicited peritoneal MΦ (TPM). In both cells, PR8 virus infection induced iNOS mRNA between 4 hr and 24 hr, attaining a peak value at 12 hr. In correlation with induction of iNOS mRNA, NO amounts increased significantly from 12 to 24 hr. Moreover, this study demonstrated that ESP with the same hemagglutination titer as PR8 virus could induce iNOS mRNA and NO production, although the inductive activity of ESP was weaker than that of PR8 virus. Considering the dual role (beneficial and detrimental roles) of NO on certain inflammatory disorders and virus infections, the inductive activity of influenza virus on the iNOS‐mediated NO production independent of its infectivity might contribute to a modification of influenza virus infection.
Further Studies on the Soluble Form (Gs) of Rabies Virus Glycoprotein (G): Molecular Structure of Gs Protein and Possible Mechanism of the Shedding
Authors: Thirapanmethee Krit , Ootaki Naohiro , Sakai Mai , Lien Chah Keng , Kawai Akihiko 
Published: November 2013

AbstractIn this study, we investigated the antigenic structures and maturation of some C‐terminal‐deficient derivatives of rabies virus glycoprotein (G). The Gs protein, a soluble form of G protein shed from infected cells, displayed antigenicity to most of our conformational epitope‐specific anti‐G mAbs, but took the 1‐30‐44 epitope‐deficient conformation (termed GC form). (The 1‐30‐44 epitope was acid‐sensitive and dependent on two separate regions, the Lys‐202‐containing and Asn‐336‐containing regions; Kankanamge et al., Microbiol. Immunol., 47: 507–519). Intact G proteins took the 1‐30‐44 epitope‐positive form (referred to as GB form) on the cell surface, but not inside the cell. A deletion mutant G(1–429) (termed GATC), lacking the transmembrane (TM) and cytoplasmic domains, was shown to be accumulated in the rough endoplasmic reticulum (rER) with BiP and did not seem to be shed. Another C‐terminal‐deficient mutant G(1–462) (termed CT1) was deprived of the whole cytoplasmic domain except for a basic amino acid left at the C‐terminus, but was transported to the cell surface, where it showed pH‐dependent cell fusion activity and almost full antigenicity to most of the anti‐G mAbs with the exception of very weak antigenicity to mAb #1‐30‐44. No Gs protein could be detected in the CT1‐producing cultures. Based on these results, we think that the cytoplasmic domain was not necessary for the G protein to be transported to the cell surface, but was necessary to keep its 1‐30‐44 epitope‐positive GB conformation. Gs proteins might have lost the C‐terminal regions during the maturation process after being exported from the rER.
Structural Difference Recognized by a Monoclonal Antibody #404‐11 between the Rabies Virus Nucleocapsid (NC) Produced in Virus Infected Cells and the NC‐Like Structures Produced in the Nucleoprotein (N) cDNA‐Transfected Cells
Authors: Toriumi Harufusa , Kawai Akihiko 
Published: November 2013

AbstractWe investigated structural changes in the rabies virus (HEP‐Flury strain) nucleocapsid (NC) during the virus replication, for which we used two anti‐nucleoprotein (N) monoclonal antibodies (mAbs), #404‐11 (specific for a conformation‐dependently exposed linear epitope) and #1‐7‐11 (specific for a conformational epitope which is exposed after the nucleocapsid formation). Both mAbs recognized the N protein of the viral NC, but not of the RNA‐free N‐P complex. The 1‐7‐11 and 404–11 epitopes could be mapped to the N‐terminal and the C‐terminal regions of N protein, respectively. Immunoprecipitation studies demonstrated that treatment of the NC either with the alkaline phosphatase or sodium deoxycholate (DOC) resulted in dissociation of most P proteins from the NC and in the reduced reactivity to mAb #404‐11, but not to mAb #1‐7‐11. NC‐like structures produced in the N cDNA‐transfected cells displayed strong reactivity to mAb #1‐7‐11; however, reactivity to mAb #404‐11 was very weak. And, co‐expression with viral phosphoprotein (P) resulted in little increase in reactivity to mAb #404‐11 of the NC‐like structures, while the reactivity was significantly increased by cotransfection with P and the viral minigenome whose 3′‐ and 5′‐end structures were derived from the viral genome. From these results, we assume that, although the 404–11 epitope is a linear one, the epitope‐containing region is exposed only when N proteins encapsidate properly the viral RNA in collaboration with the P protein. Further, exposure of the 404–11 epitope region might be function‐related, and be regulated by association and dissociation of the P protein.
Organization of the CTX Prophage in Environmental Isolates of Vibrio mimicus
Authors: Islam Mohammad Sirajul , Rahman Mohammed Ziaur , Khan Sirajul Islam , Mahmud Zahid Hayat , Ramamurthy Thandavarayan , Nair Gopinath Balakrish , Sack Richard Bradley , Sack David Allen 
Published: November 2013

AbstractThe organization of the CTX prophage in environmental strains of Vibrio mimicus was investigated. Sixteen hundred non‐sucrose fermenting vibrios were examined for ctx gene by hybridization. Out of 1,600 isolates, 6 V. mimicus isolates contained ctxA gene. The organization of CTX prophage was determined by RFLP using ctxA probe. The CTX prophage integrated at a single site in V. mimicus genome which was present as a single copy flanked by at least a single RS element. Ribotype pattern revealed that a particular clone of V. mimicus acquired the CTXφ in the aquatic environment. This study demonstrated that V. mimicus could act as a reservoir of CTXφ in the aquatic environment.
Effects of Type II Alveolar Epithelial Cells on T Cell Mitogenic Responses to Concanavalin A and Purified Protein Derivatives
Authors: Sato Katsumasa , Shimizu Toshiaki , Sano Chiaki , Tomioka Haruaki 
Published: November 2013

AbstractTo investigate the role of type II alveolar epithelial cells during the T cell‐dependent host immune response to Mycobacterium tuberculosis (MTB), effects of MTB‐infected A‐549 human type II alveolar epithelial cells (A‐549 cells) on T cell mitogenesis in response to concanavalin A (Con A) and purified protein derivatives (PPD) were studied. Human peripheral blood mononuclear cells (PBMCs) were co‐cultivated with uninfected or MTB‐infected A‐549 cells and Con A‐ and PPD‐induced T cell mitogeneses were examined, and the following findings were obtained. T cell mitogenesis was inhibited by uninfected as well as MTB‐infected A‐549 cells, even when a dual‐chamber culture system was used to prevent direct cell contact between PBMCs and A‐549 cells. Therefore, it appears that A‐549 cells suppress T cell mitogenesis by producing some unknown humoral suppressor factors.
In Silico Analysis of ORF1ab in Coronavirus HKU1 Genome Reveals a Unique Putative Cleavage Site of Coronavirus HKU1 3C‐Like Protease
Authors: Woo Patrick C.Y. , Huang Yi , Lau Susanna K.P. , Tsoi Hoi‐wah , Yuen Kwok‐yung 
Published: November 2013

AbstractRecently we have described the discovery and complete genome sequence of a novel coronavirus associated with pneumonia, coronavirus HKU1 (CoV‐HKU1). In this study, a detailed in silico analysis of the ORF1ab, encoding the 7,182‐amino acid replicase polyprotein in the CoV‐HKU1 genome showed that the replicase polyprotein of CoV‐HKU1 is cleaved by its papain‐like proteases and 3C‐like protease (3CLpro) into 16 polypeptides homologous to the corresponding polypeptides in other coronaviruses. Surprisingly, analysis of the putative cleavage sites of the 3CLpro revealed a unique putative cleavage site. In all known coronaviruses, the P1 positions at the cleavage sites of the 3CLpro are occupied by glutamine. This is also observed in CoV‐HKU1, except for one site at the junction between nsp10 (helicase) and nsp11 (member of exonuclease family), where the P1 position is occupied by histidine. This amino acid substitution is due to a single nucleotide mutation in the CoV‐HKU1 genome, CAG/A to CAT. This probably represents a novel cleavage site because the same mutation was consistently observed in CoV‐HKU1 sequences from multiple specimens of different patients; the P2 and P1′‐P12′ positions of this cleavage site are consistent between CoV‐HKU1 and other coronaviruses; and as the helicase is one of the most conserved proteins in coronaviruses, cleavage between nsp10 and nsp11 should be an essential step for the generation of the mature functional helicase. Experiments, including purification and C‐terminal amino acid sequencing of the CoV‐HKU1 helicase and trans‐cleavage assays of the CoV‐HKU1 3CLpro will confirm the presence of this novel cleavage site.
Legionella pneumophila Serogroup 1 Isolates from Cooling Towers in Japan Form a Distinct Genetic Cluster
Authors: Amemura‐Maekawa Junko , Kura Fumiaki , Chang Bin , Watanabe Haruo 
Published: November 2013

AbstractThirty‐one epidemiologically unrelated Legionella pneumophila serogroup 1 isolates (10 from cooling towers, 10 from public spas and/or hot spring baths, and 11 from patients) were analyzed by pulsed‐field gel electrophoresis (PFGE) and sequence‐based typing (SBT) using 6 loci, flaA, pilE, asd, mip, mompS, and proA. The results of PFGE and SBT analysis indicated that all 10 isolates from cooling towers clustered into a unique type, which was distinct from strains of other environmental sources.
Increased Expression of c‐myc Is Associated with Thymoma in Rats Infected with Murine Leukemia Virus A8
Authors: Hirano Tomoko , Watanabe Rihito , Takase‐Yoden Sayaka 
Published: November 2013

AbstractInfection of rats with Friend murine leukemia virus (Fr‐MLV) clone A8 causes thymoma in all the animals within 7 weeks. The rapid induction of thymoma is associated with a unique enhancer structure in the U3 region of the A8‐LTR. Our Southern blot analyses showed that the thymomas were oligo clonal. The A8‐induced thymomas showed 3‐ to 11‐fold overexpression of c‐myc mRNA. These results suggest that provirus insertion into particular positions of the host genome is correlated with tumorigenesis after A8 infection and that up‐regulation of c‐myc plays an important role in the induction of thymoma.
Production of Autoantibodies Associated with Polyclonal Activation in Yersinia enterocolitica O:8‐Infected Mice
Authors: Ramos Orivaldo Pereira , Silva Eloisa Elena Cangiani , Falcão Deise Pasetto , de Medeiros Beatriz Maria Machado 
Published: November 2013

AbstractPolyclonal lymphocyte stimulation is one of the immunomodulatory mechanisms induced by arthritogenic pathogens. In this study we examined the polyclonal activation potential of a virulent strain of Y. enterocolitica serotype O:8 (WA 2707+) and its plasmidless isogenic pair (WA 2707−). SPF Swiss mice were infected intragastrically and spleen cells were obtained on days 7, 14, 21, 28, 35 and 42 after infection. The number of cells secreting nonspecific immunoglobulins of IgG, IgM and IgA isotypes was determined by the ELISPOT technique. The presence of serum‐specific antibodies was investigated by ELISA and the presence of autoantibodies by dot‐blot assay. Although the patterns of infection of the two bacterial strains were almost the same, only the animals infected with the virulent strain presented clinical anomalies. Neither arthritic nor inflammatory signs were observed in the joints of the infected animals. The greatest activation observed was that of the nonspecific IgM‐secreting cells, and their peak of secretion occurred between the 28th and the 42nd day after infection, for both strains of Y. enterocolitica O:8. Only the animals infected with the virulent strain (WA 2707+) produced IgG‐specific antibodies in the serum, from the 28th day after infection. The serum of animals infected with either strain showed reactivity to all the autologous constituents tested, mainly on the 28th and 42nd day after infection. It was concluded that infection of mice with either the virulent strain of Y. enterocolitica O:8 or with its plasmidless isogenic pair resulted in the polyclonal activation of the splenic B lymphocytes including some autoreactive clones.
Quantification of Aquabirnaviruses Isolated from Different Host Species by Real‐Time RT‐PCR
Authors: Hirayama Takeshi , Imajoh Masayuki , Oshima Syun‐ichirou 
Published: November 2013

AbstractReal‐time reverse transcription (RT)‐PCR was developed to detect and quantify successfully aquatic birnaviruses (ABVs) from fish tissue and seawater samples. This method detected marine birnavirus (MABV) RNA in several samples, and the detection was specific for ABVs. Monitoring the MABV strain Y‐6 RNA quantification by real‐time RT‐PCR showed replication kinetics of MABV after experimental infection in vitro. We found the quantity of ABVs isolated from different host species by using combined virus absorption in cell culture and real‐time RT‐PCR. Although all specimens showed no symptoms of viral infection, ABVs were detected regardless of host species. In conclusion, real‐time RT‐PCR was shown to be a sensitive and reliable tool to detect and quantify ABVs in cultured fish. This method is useful procedure to show details of horizontal or vertical infections by ABVs in the breeding water of aquatic animals.
Practical Range of Effective Dose for Cre Recombinase‐Expressing Recombinant Adenovirus without Cell Toxicity in Mammalian Cells
Authors: Baba Yasuko , Nakano Masakazu , Yamada Yoshitsugu , Saito Izumu , Kanegae Yumi 
Published: November 2013

AbstractThe site‐specific recombinase Cre is valuable for regulation of gene expression not only in vitro but also in vivo. We previously reported that replication‐deficient recombinant adenovirus (rAd) expressing Cre can mediate efficient and strict regulation in 100% of cultured cells. Recently, the constitutive‐expression of Cre using retrovirus or lentivirus vector reportedly inhibited cell‐growth, but the effect of transient Cre expression have not yet been examined. Here we showed that an excess amount of Cre produced from Cre‐expressing rAd caused a deleterious effect in cells even when Cre was transiently expressed. We used three rAds carrying promoters with different activities: the SV40 early promoter (AxSVENCre), the SRα promoter (AxSRCre) and the CAG promoter (AxCANCre). Cell toxicity was clearly caused by Cre itself and was distinguishable from that caused by rAd virions when the cytopathic effects of these rAds were compared with that of a control virus lacking the Cre expression unit. Cre toxicity was strongly correlated with the expression level of Cre. Importantly, AxSRCre and AxCANCre gave a 60‐fold range of effective MOIs (“effective range”) sufficient for gene activation without causing cell toxicity from either the rAd particles or Cre itself, while AxSVENCre failed to give such a range because the expression level of Cre was too low. When Cre was tagged with a nuclear localization signal (NLS), not only its activity but also Cre toxicity was increased fourfold, and the effective range was unchanged. Therefore, AxSRNCre might be more useful to control cell toxicity from the rAd virions than AxSRCre. Cre‐induced cell toxicity can be avoided by pre‐examining the “effective range” using the purpose cell lines before starting experiments utilizing the experiment of Cre‐expressing rAd.
Application of Real‐Time PCR for Quantitative Detection of Clostridium botulinum Type A Toxin Gene in Food
Authors: Yoon So‐Yeon , Chung Gyung Tae , Kang Do‐Hyun , Ryu Chunsun , Yoo Cheon‐Kwon , Seong Won‐Keun 
Published: November 2013

AbstractThe TaqMan real‐time PCR method for the quantitative detection of C. botulinum type A was developed based on sequence‐specific hybridization probes. The validity of this assay was verified by using 10 genera of 20 strains, including reference strains of C. botulinum types A, B, C, D, E and F. The detection limit of this assay was evaluated on C. botulinum type A, using a 10‐fold dilution series of DNA and spores. The DNA and spores were detected up to level of 0.1 ng/ml and 102 spores/ml, respectively. Spore spiked food sample preparation prior to the real‐time PCR was performed by two methods, heat treatment and GuSCN. The detection limits after heat treatment showed 102 spores/ml for spiked sausage slurry, and 103 spores/ml for spiked canned corn slurry, while detection limits after GuSCN precipitation showed 102 spores/ml in both sausage and canned corn. Therefore the real‐time PCR assay after GuSCN precipitation is useful for the quantification of C. botulinum type A because it showed identical CT values in both pure spore solutions and food slurries. We suggest that quantitative analysis of C. botulinum type A by TaqMan real‐time PCR can be a rapid and accurate assessment method for botulinal risk in food samples.
Highly Pathogenic H5N1 Influenza Virus Causes Coagulopathy in Chickens
Authors: Muramoto Yukiko , Ozaki Hiroichi , Takada Ayato , Park Chun‐Ho , Sunden Yuji , Umemura Takashi , Kawaoka Yoshihiro , Matsuda Haruo , Kida Hiroshi 
Published: November 2013

AbstractSevere hemorrhage at multiple organs is frequently observed in chickens infected with highly pathogenic avian influenza (HPAI) A viruses. In this study we examined whether HPAI virus infection leads to coagulation disorder in chickens. Pathological examinations showed that the fibrin thrombi were formed in arterioles at the lung, associated with the viral antigens in endothelial cells of chickens infected intravenously with HPAI virus. Hematological analyses of peripheral blood collected from the chickens revealed that coagulopathy was initiated at early stage of infection when viral antigens were detected only in the endothelial cells and monocytes/macrophages. Furthermore, gene expression of the tissue factor, the main initiator of blood coagulation, was upregulated in the spleen, lung, and brain of HPAI virus‐infected chickens. These results suggest that dysfunction of endothelial cells and monocytes/macrophages upon HPAI virus infection may induce hemostasis abnormalities represented by the excessive blood coagulation and consumptive coagulopathy in chickens.
Correlation between the Anti‐Virus‐Induced Cytopathic Effect Activity of Interferon‐α Subtypes and Induction of MxA Protein In Vitro
Authors: Schanen Cécile , Chieux Vincent , Lobert Pierre‐Emmanuel , Harvey Jeanne , Hober Didier 
Published: November 2013

AbstractThere are several interferon‐α (IFN‐α) subtypes. Mechanism of disparity in biological effects among members of IFN‐α subtypes remains unexplained. Biological activity of IFN‐α is mediated in part by induction of intracellular antiviral proteins. We studied whether differences in biologic effects of IFN‐α subtypes may rely on their antiviral protein inducing effect. Intracellular induction of MxA protein and anti‐virus‐induced cytopathic effect (CPE) activity of 11 IFN‐α subtypes in human amnion WISH cells have been studied. MxA protein quantitation in cell lysates was performed by immunochemiluminescence assay and anti‐virus‐induced CPE activity was assessed by protection against vesicular stomatitis virus (VSV)‐induced CPE. Range of MxA values was high when cells were treated with 10 and 100 IU/ml of each IFN‐α subtype. Levels of MxA correlated with anti‐VSV‐induced CPE obtained with 10 IU/ml IFN‐α subtype. Together our data show a disparity in MxA‐inducing activity of IFN‐α subtypes and suggest that differences in anti‐VSV‐induced CPE of IFN‐α subtypes in WISH cells can be related to their different ability to induce MxA.