bluuebunny/crossref_metadata_2025_split huggingface dataset:
Studies on the Rabies Virus RNA Polymerase: 2. Possible Relationships between the Two Forms of the Non‐Catalytic Subunit (P Protein)
Authors: Takamatsu Fumihiko , Asakawa Naoki , Morimoto Kinjiro , Takeuchs Kenji , Eriguchi Yoshiro , Toriumi Harufusa , Kawai Akihiko 
Published: November 2013

AbstractWe investigated the relationship between the two forms of rabies virus P protein, a non‐catalytic subunit of rabies virus RNA polymerase. The two displayed different electrophoretic mobilities as 37‐ and 40‐kDa polypeptides, hence termed as p37 and p40, respectively. Double labeling experiments with [3H]leucine and [32P]orthophosphate demonstrated that p40 was much more phosphorylated than p37. Treatment of the virion proteins with alkaline phosphatase eliminated only p40, and not 37‐kDa polypeptide. The p37 was a major product of the P gene, and was accumulated in the infected cell and incorporated into the virion. On the other hand, p40 was apparently detected only in the virion, and little detected in the cells. Treatment of infected cells with okadaic acid, however, resulted in significant accumulation of p40 in the cell, suggesting that p40 was continuously produced in the cell but dephosphorylated quickly. We detected both 37‐ and 40‐kDa products in P cDNA‐transfected animal cells, while only a 37‐kDa product was produced in Escherichia coli. Incubation of 37‐kDa products from E. coli with the lysates of animal cells in vitro resulted in the production of a 40‐kDa product, which was also shown to be suppressed by the heparin. From these results, it is suggested that p40 is produced by the hyperphosphorylation of a 37‐kDa polypeptide, which depends on certain heparin‐sensitive cellular enzyme(s) and occurs even in the absence of the other viral gene products, and that p40 is reverted quickly to p37 in the infected cells, probably being dependent on some virus‐induced factor(s).
Detection and Genetical Characterization of Shiga Toxin‐Producing Escherichia coli from Wild Deer
Authors: Asakura Hiroshi , Makino Sou‐ichi , Shirahata Toshikazu , Tsukamoto Teizo , Kurazono Hisao , Ikeda Tetsuya , Takeshi Kouichi 
Published: November 2013

AbstractShiga toxin (Stx)‐producing Escherichia coli (STEC) strains isolated from wild deer in Japan were examined. A total of 43 fecal samples were collected 4 times from 4 different sites around Obihiro City, Hokkaido, Japan, in June and July 1997. Seven STEC strains were isolated by PCR screening, all of them were confirmed by ELISA and Vero cell cytotoxicity assay to be producing only active Stx type 2 (Stx2). Moreover, they seemed to carry the hemolysin and eaeA genes of STEC O157:H7, and some isolates harbored large plasmids which were similar to the 90‐kilobase virulence plasmid of STEC O157:H7. Based on their plasmid profiles, antibiotic resistance patterns, PCR‐based DNA fingerprinting data obtained by using random amplified polymorphic DNA (RAPD) and the stx2 gene sequences, all isolates were divergent from each other except for 3 isolates from the first and second samplings. A DNA sequence analysis of representative isolates revealed that deer originating STEC strains were closely related to each other, but not to the Stx2‐producing STEC strains isolated from a mass outbreak in Obihiro at the same time. A phylogenic analysis of the deduced Stx2 amino acid sequences demonstrated that three distinct clusters existed in the deer originating STEC strains and that the Stx of deer originating STEC was closely associated with that originating from humans, but not those of STEC originating from other animals. These results suggest that STEC contamination of deer carcasses should be considered as a potential source of human infection and adequate sanitary inspection of meat for human consumption is also essential for wild animals.
Identification and Semiquantitation of Mycobacterium avium Using a Competitive PCR Method
Authors: Hashimoto Toru , Suzuki Katsuhiro 
Published: November 2013

AbstractA competitive PCR method with standard DNA (MIMIC) was developed for the rapid detection and semiquantitation of Mycobacterium avium (M. avium) using primers specific for the alpha antigen sequence of the bacteria. DNA from both M. avium and Mycobacterium marinum was amplified by polymerase chain reaction (PCR), but only M. avium could be detected by subsequent blotting confirmation with a probe specific for the bacteria. With the PCR and subsequent dot blot hybridization, as little as 10 fg of the M. avium DNA could be detected, equivalent to about 2 cells of the mycobacteria. In addition, we could distinguish 105 CFUs of M. avium from 104 CFUs or less by competitive PCR using a MIMIC. The present competitive PCR test enabled rapid identification and semiquantitation of M. avium, and could be used clinically to monitor disease severity and response to treatment of human M. avium disease.
Specific Microbial Colonizations in the Periodontal Sites of HIV‐infected Subjects
Authors: Chattin Blanca Rosa , Ishihara Kazuyuki , Okuda Katsuji , Hirai Yoshito , Ishikawa Tatsuya 
Published: November 2013

AbstractThe purpose of this study was to examine colonization by specific organisms at periodontal sites in HIV‐seropositive [HIV(+)] subjects. A total of 67 HIV(+) and 32 HIV(‐) subjects were investigated. The specific pathogens included black‐pigmented anaerobic rods (BPAR), Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Candida albicans and mycoplasma species. P. gingivalis was present in the HIV(+) subjects more frequently than in the HIV(‐) periodontitis patients (P < 0.01). The cell numbers of BPAR and P. gingivalis and percentages as the total of CFUs on blood agar cultured in an anaerobic chamber were statistically higher in periodontal pocket samples from HIV(+) than from HIV(‐). A. actinomycetemcomitans was also detected at a high rate (41.8%) in HIV(+) patients. The average cell numbers of C. albicans were higher in samples from the HIV(+) group (P < 0.05). The detection rate of mycoplasma species in the HIV(+) patients was significantly lower than that in HIV(‐) subjects (P < 0.05), and most isolated mycoplasma strains were Mycoplasma salivarium.
Siderophore Production of Vibrio parahaemolyticus Strains from Different Sources
Authors: Yamamoto Shigeo , Okujo Noriyuki , Miyoshi Shin‐Ichi , Shinoda Sumio , Narimatsu Shizuo 
Published: November 2013

AbstractVibrio parahaemolyticus strains isolated from different sources were assayed for their ability to produce a siderophore, vibrioferrin, under iron‐limited growth conditions. The mean value } standard error of mean μM vibrioferrin in spent culture supernatant/optical density at 660 nm) was 832.3 } 66.9 for clinical isolates (n 44), which was significantly higher (P 0.01) than those for food isolates (461.0 } 66.5; n 37) and coastal isolates (378.8 } 37.2; n 26). This suggests that greater productivity of vibrioferrin by clinical isolates may be associated with a selective advantage for survival and proliferation under conditions of iron‐limitation such as in the intestine.
Partial Purification and Characterization of the Active Entity Responsible for Inducing Interleukin‐6 Production by Human Gingival Fibroblasts from Mycoplasma salivarium Cells
Authors: Hasebe Akira , Shibata Ken‐Ichiro , Domon Hikouji , Dong Li , Watanabe Tsuguo 
Published: November 2013

AbstractThe active entity responsible for inducing interleukin‐6 production by human gingival fibroblasts was partially purified by ion‐exchange chromatography from the water‐soluble fraction of Mycoplasma salivarium cells. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the final preparation revealed one densely stained band with a molecular weight of 20.6 kilodaltons and two faint bands with molecular weights of 40.5 and 82.5 kilodaltons. The specific activity of the final preparation was 34‐fold higher than that of the starting water‐soluble fraction. The interleukin‐6‐inducing activity was destroyed by proteinase K and reduced 70% by lipoprotein lipase and heat treatment, but was not affected by deoxyri‐bonuclease I or endoglucosidase D. The final preparation induced small amounts of tumor necrosis factore‐ α and interleukin‐1β in a myelomonocytic cell line, THP‐1 cells, but did not induce interleukin‐6. The ability of Escherichia coli lipopolysaccharide to stimulate human gingival fibroblasts to release interleukin‐6 was dependent upon the presence of serum in the assay medium, but that of the final preparation from M. salivarium was not. Thus, we partially purified the protein(s) from M. salivarium which were capable of stimulating human gingival fibroblasts to release interleukin‐6 by a mechanism different from that of E. coli lipopolysaccharide.
Adenovirus Type 5 E1A Immortalizes Primary Rat Cells Expressing Wild‐Type p53
Authors: Yamashita Toshiharu , Tonoki Hidefumi , Nakata Daichi , Yamano Shigeru , Segawa Kaoru , Moriuchi Tetsuya 
Published: November 2013

AbstractAdenovirus (Ad) E1A induces apoptosis in cells expressing wild‐type p53, and stable transformation by Ad E1A requires the co‐introduction of an anti‐apoptotic gene such as Ad E1B 19K. Thus, cells immortalized by Ad E1A alone might have lost functional p53. In order to analyze the p53 in rat cells expressing Ad E1A, we established rat cell lines by transfecting primary rat embryo fibroblast (REF) and baby rat kidney (BRK) cells with cloned Ad5 E1A. By using a yeast functional assay, we analyzed p53 in six primary REF and three BRK cell lines immortalized by Ad5 E1A as well as five spontaneously immortalized rat cell lines (REF52, NRK, WFB, Rat‐1 and 3Y1). The yeast functional assay revealed that all of the spontaneously and Ad5 E1A‐immortalized rat cell lines except for 3Y1 expressed wild‐type p53. All of the Ad5 E1A‐immortalized rat cell lines contained p53 detectable by immunoprecipitation. Recombinant adenovirus expressing rat p53 cloned from a REF cell line immortalized by Ad5 E1A, as well as that expressing murine wild‐type p53, induced apoptosis in p53‐null cells in collaboration with E1A. Thus, it is suggested that the mutation of p53 appears to be not frequent in the spontaneous immortalization of primary rat cells, and that the functional loss of wild‐type p53 is not a prerequisite of E1A‐mediated immortalization.
The hsp Operons Are Repressed by the hrc37 of the hsp70 Operon in Staphylococcus aureus
Authors: Kuroda Makoto , Kobayashi Daisuke , Honda Kyoko , Hayashi Hideo , Ohta Toshiko 
Published: November 2013

AbstractThe heat‐shock proteins are coded for the polycistronic operons hsp70 and hsp60 in Staphylococcus aureus. The hsp70 operon is comprised of five genes, hrc37, hsp20, hsp70, hsp40 and orf35, and the hsp60 is comprised of two genes, hsp10 and hsp60. The hsp70 operon transcribed five different sizes of mRNA from three promoters: P1, the most active promoter, transcribed 6.0 and 3.6 kb mRNAs; P2 transcribed a single 1.8 kb mRNA; and P3 transcribed 4.2 and 2.4 kb mRNAs. The hsp60 operon transcribed a single 2.1 kb transcript from only one promoter, P1. Both operons had a common structure of inverted repeat element (CIRCE, Controlling Inverted Repeat of Chaperon Expression) at the promoter region. All of the transcripts were heat (46 C) inducible. One of the unidentified genes, hrc37, was characterized. The disruptant of the hrc37 in the hsp70 operon enhanced the transcription of both operons at 37 C (derepression). Complementation of the disruptant with the cloned hrc37 plasmid recovered the repression of the transcription of both operons at 37 C. The product of hrc37, Hrc37, was found to bind to the CIRCE element. These findings indicated that Hrc37 from the hsp70 operon repressed the transcription of both the hsp70 and hsp60 operons by binding to the CIRCE element located at the promoter region.
The Hemagglutinating Action of Vibrio vulnificus Metalloprotease
Authors: Miyoshi Shin‐ichi , Kawata Koji , Tomochika Ken‐ichi , Shinoda Sumio 
Published: November 2013

AbstractVibrio vulnificus protease (VVP), a 45‐kDa zinc metalloprotease, consists of two functional domains: an N‐terminal 35‐kDa polypeptide having endoproteinase activity, and a C‐terminal 10‐kDa polypeptide that mediates the binding of VVP to the erythrocyte membrane. Therefore, VVP, but not its N‐terminal endoproteinase domain alone, has agglutinating activity to rabbit erythrocytes. When a single zinc atom in the catalytic center was substituted by treatment with CuCl2 or NiCl2, proteolytic and hemagglutinating activities were reduced by Ni substitution but not by Cu substitution. Cu‐treated 35‐kDa polypeptide showed sufficient affinity of the catalytic center and weak binding ability to the erythrocyte membrane, but the Ni‐treated polypeptide did not. These results suggest that the binding of endoproteinase domain to membrane is also necessary for hemagglutination.
Expression of Lymphotoxin Gene Inserted into Theiler's Murine Encephalomyelitis Virus
Authors: Obuchi Masatsugu , Odagiri Takato , Lizuka Hideaki , Ohara Yoshiro 
Published: November 2013

AbstractTheiler's murine encephalomyelitis virus (TMEV) belongs to the Picornaviridae genus. DA subgroup strains of this virus induce early, non‐fatal polioencephalomyelitis followed by demyelination in the spinal cord, with virus persistence. We investigated the use of DA strain as a vector for the introduction of a foreign gene into the central nervous system. Human lymphotoxin (LT) gene was inserted in the L region, the most upstream part of the polyprotein coding region of DA genome. Expression of LT was demonstrated by an immunoblot and an enzyme‐linked immunosorbent assay on BHK‐21 cells that were infected with the recombinant virus. In addition, the expressed LT showed cytotoxicity against L‐929 cells.
Protective Immunity Induced by Vaccination with SAG1 Gene‐Transfected Cells against Toxoplasma gondii‐Infect ion in Mice
Authors: Aosai Fumie , Mun Hye‐seong , Norose Kazumi , Chen Mei , Hata Hidekazu , Kobayashi Masashi , Kiuchi Masahiro , Stauss Hans J. , Yano Akihiko 
Published: November 2013

AbstractTo develop a vaccine by augmenting the protective cellular immunity against Toxoplasma gondii (T. gondii)‐infection, T. gondii SAG1 gene‐transfectants were established by using RMA.S (H‐2b), a murine transporter associated with the antigen processing (TAP) molecule‐deficient lymphoma line, as a host antigen‐presenting cell (APC). Immunization of C57BL/6 mice with the SAG1‐transfected RMA.S induced CD8+ cytotoxic T lymphocytes (CTL) specific for not only SAG1‐transfected RMA.S but also T. gondii‐infected RMA.S, and elicited protective responses to infection with a virulent T. gondii strain, RH.
The Effects of S‐Carboxymethylcysteine and N‐Acetylcysteine on the Adherence of Moraxella catarrhalis to Human Pharyngeal Epithelial Cells
Authors: Zheng Can Hong , Ahmed Kamruddin , Rikitomi Naoto , Martinez Glenda , Nagatake Tsuyoshi 
Published: November 2013

AbstractWe investigated the effects of two mucoregulating drugs, S‐carboxymethyleysteine (S‐CMC) and N‐acetylcysteine (NAC), on the attachment of Moraxella catarrhalis (M. catarrhalis) to pharyngeal epithelial cells. The attachment of M. catarrhalis decreased (3357%) significantly (P<0.01) in a dose‐dependent manner in cells treated with mucoregulating drugs as compared to the control. There was a significant (P< 0.01) decrease (35 45%) in the attachment of M. catarrhalis to pharyngeal cells after oral administration of S‐CMC. By electron microscopic observation, it was found that there was a fine, granular, electron‐dense, ruthenium red‐positive layer on the surface of pharyngeal epithelial cells; this layer was absent on cell surfaces treated with mucoregulating drugs. Possibly, this layer contained the portion of M. catarrhalis receptor which is responsible for the attachment of this bacteria to pharyngeal epithelial cells. From the above results, it may be concluded that one of the mechanisms of mucoregulating drugs to decrease the episode of respiratory infections in patients with chronic respiratory diseases is by inhibiting the attachment of bacteria to the upper respiratory tract.
Genetic Relationships among Mycoplasmas Based on the 16S‐23S rRNA Spacer Sequence
Authors: Harasawa Ryô 
Published: November 2013

AbstractThe nucleotide sequences of the spacer regions between the 16S and 23S rRNA genes of 20 Mycoplasma species were determined following amplification by PCR. Although the spacer regions lacked spacer tRNA genes, they contained the box B and box A sequences in this order from the 5′ terminus. The sequence alignment indicated that the 20 species were divided into four clusters, the M. pneumoniae, M. hominis, M. hyorhinis and M. fermentans clusters, and a single floating species, M. hyopneumoniae.
Genetic Variation in the VP4 and NSP4 Genes of Human Rotavirus Serotype 3 (G3 Type) Isolated in China and Japan
Authors: Cao Xiu‐ru , Akihara Shiho , Fang Zhao‐yin , Nakagomi Osamu , Ushijima Hiroshi 
Published: November 2013

AbstractSequence analyses of the VP4 and NSP4 genes were performed on twenty human isolates of serotype G3 rotavirus obtained from China and Japan. One isolate from China, CHW17, possessed P[4] genotype VP4 and KUN group NSP4 genes which are associated with G2. One isolate (02/92) from Japan, which was shown to have a wider spacing between RNA segments 10 and 11 by RNA polyacrylamide gel electrophoretic analysis like AU‐1, possessed P[9] genotype VP4 and AU‐1 group NSP4 genes. The other isolates had P[8] genotype VP4 and Wa group NSP4 genes. While the nucleotide sequence conservation among the G3 VP7 genes was more than 79% (Wen et al, Arch. Virol., 1997, 142: 1481 1489), the conservation of VP4 and NSP4 genes in the same genotypes or groups was more than 85%.
High Molecular Weight Dextran Sulfate Increases the Activity of NF‐κB‐Regulated Promoter in Monocyte‐Derived Macrophages
Authors: Jagodzinski Pawel P. , Trzeciak Wiesław H. 
Published: November 2013

AbstractIt is known that sulfated polysaccharides can mimic the action of common T‐cell mitogens. To investigate the molecular basis of the mitogenic effect of high molecular weight dextran sulfate (HMDS), monocyte‐derived macrophages were transfected with recombinant plasmid containing chloramphenicol acetyl transferase (CAT) reporter gene under the control of the HIV‐1 long terminal repeat (LTR) promoter, which is regulated by transcription factor NF‐κB. We observed that HMDS, similar to bacterial lipopolysaccharide (LPS), increases the expression of CAT reporter gene suggesting increased activity of NF‐κB. The activation of NF‐κB correlated with the increased expression of B7.1 molecules. It was postulated that this NF‐κB‐regulated promoter might play a role in the activation of the accessory cells as well as the rate of replication of HIV‐1 in monocyte‐derived macrophages.
A Novel Negative Regulator Molecule, Cho‐1, Is Involved in the Cytotoxicity by Human Natural Killer Cells but Not in Cytotoxic T Lymphocytes
Authors: Akazawa Takashi , Hirai Itaru , Hirohashi Yoshihiko , Kamiguchi Kenjiro , Sahara Hiroeki , Torigoe Toshihiko , Nagasawa Shigeharu , Tamura Yasuaki , Sato Noriyuki 
Published: November 2013

AbstractWe previously reported the cytotoxic negative regulatory molecule, Cho‐1, that was expressed on the cell surface of rat fetal fibroblast cells in the cytotoxicity by natural killer (NK) cells. This molecule was IFN‐γ‐inducible, but appeared to be different from MHC class I. It was expressed on NK‐resistant cells but not on NK‐sensitive murine target cells such as YAC‐1. In this paper, first we determined whether Cho‐1 could also act as the negative regulatory molecule in a human NK‐resistant HEPM line. Our data strongly suggested that Cho‐1 could act as such a negative regulatory molecule in human NK cytotoxicity. The immunoprecipitates made with HEPM cell lysate and anti‐MHC class I monoclonal antibody (mAb) did not react against anti‐Cho‐1 mAb, indicating that Cho‐1 was different from MHC class I. Second, an assessment was made as to whether or not this molecule is involved in the cytotoxicity of CD8 (+) cytotoxic T lymphocytes (CTL) against human autologous tumor cells. The data indicated that although this cell surface molecule was expressed on certain tumor lines, it was not involved in the cytotoxic mechanism of CTL. Thus, Cho‐1 appeared to be the novel regulatory molecule in the NK cytotoxic mechanism.
Disappearance of Glyceraldehyde 3‐Phosphate Dehydrogenase from Erythrocyte Membrane by Hemolysis with Thermostable Direct Hemolysin of Vibrio parahaemolyticus or Vibrio cholerae El Tor Hemolysin
Authors: Zhang Dongyan , Honda Takeshi 
Published: November 2013

AbstractIt is believed that the thermostable direct hemolysin (TDH) of Vibrio parahaemolyticus and El Tor hemolysin (ETH) of Vibrio cholerae damage erythrocytes and other cells by acting as pore‐forming toxins. In this study, we found that a protein band with a molecular weight of 37,000 daltons specifically disappeared from erythrocyte membrane after hemolysis by TDH and ETH, but not after hypotonic hemolysis. The 37 kDa band was identified as glyceraldehyde 3‐phosphate dehydrogenase (G3PD), a glycolytic enzyme, based on N‐terminal 14 amino acid sequencing. The role of G3PD in hemolysis is discussed.
In Vitro Assessment of a Chemically Synthesized Shiga Toxin Receptor Analog Attached to Chromosorb P (Synsorb Pk) as a Specific Absorbing Agent of Shiga Toxin 1 and 2
Authors: Takeda Tae , Yoshino Ken‐ichi , Adachi Eriko , Sato Yuko , Yamagata Kyoko 
Published: November 2013

AbstractA synthetic analog of Shiga toxin (Stx) receptor (Synsorb Pk) was quantitatively assessed to determine whether it can protect human renal adenocarcinoma cells (ACHN cells) from the cytotoxicity of Stx1 and Stx2 by coincubation experiments. Coincubation of 100 and 20 ng of Stx1 and Stx2 with 50 mg of Synsorb Pk for 1 hr at 37 C in 1 ml of Eagle's Minimum Essential Medium supplemented with 1% (v/v) nonessential amino acid and 10% (v/v) fetal calf serum protected 50% of the cells from the cytotoxic effect. Chromosorb P, an inert matrix control, did not absorb the Stxs at all. Heat‐treatment (boiled for 10 min) to Synsorb Pk caused a 50% decrease in Stx2‐binding activity, but did not effect the Stx1 binding. Further, Stxs bound to Synsorb Pk could be demonstrated. When 20 mg of Synsorb Pk was coincubated for 30 min at 37 C in 1 ml of phosphate‐buffered saline with 1 and 10 ng or more of Stx1 or Stx2, respectively, the toxins could be detected on the surface when the bound toxins on Synsorb Pk were used as the solid phase in enzyme immunoassay. The amount of 100 ng/ml of both Stx1 and Stx2 appeared to saturate 20 mg/ml of Synsorb Pk after coincubating for 30 min at 37 C. While assessing the Stxs' binding activity to Synsorb Pk, it was demonstrated that Stx1 had a higher affinity to Pk trisaccharide than Stx2. These observations provide useful information on the effectiveness of Synsorb Pk to trap and eliminate free Stxs produced in the gut of patients infected by Stx‐producing Escherichia coli, and to prevent the progression of hemorrhagic colitis to hemolytic uremic syndrome.