bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of the NH2‐Terminal 87th Amino Acid of Escherichia coli GyrA in Quinolone‐Resistance
Authors: Yonezawa Minoru , Takahata Masahiro , Banzawa Naoko , Matsubara Nobuyuki , Watanabe Yasuo , Narita Hirokazu 
Published: November 2013

AbstractThe functional contributions of amino acid residue Asp87 of Escherichia coli gyrase A protein (GyrA) was analyzed by site‐directed mutagenesis. We generated a series of mutants, in which Asp87 of GyrA was changed to Ala, Val, Phe, Asn, Ser, and Lys. By genetic analysis of gyrA genes in a gyrA temperature‐sensitive (Ts) background, it was shown that all these mutations caused the quinolone‐resistance. These results indicate that the 87th amino acid of E. coli GyrA must have negative charge in expressing the phenotype of quinolone sensitivity. These findings also suggest that the carboxyl group of Asp87 may interact with quinolone drugs.
Nucleotide Sequence of the Gene Encoding β‐Isopropylmalate Dehydrogenase (leuB) from Bacteroides fragilis
Authors: Sarker Mahfuzur R. , Akimoto Shigeru , Ugai Hideyo , Kuwahara Tomomi , Ohnishi Yoshinari 
Published: November 2013

AbstractThe complete nucleotide sequence of the gene (leuB) coding for β‐isopropylmaiate dehydrogenase of Bacteroides fragilis was determined. An open reading frame of 1,061 nucleotides was detected that could encode a polypeptide of 353 amino acid residues with a calculated molecular mass of 39,179 Da. The deduced amino acid sequence of the β‐isopropylmalate dehydrogenase from B. fragilis showed substantial sequence similarity with the β‐isopropylmalate dehydrogenases from other bacteria.
Distribution of C22‐, C24‐ and C26‐α‐Unit‐Containing Mycolic Acid Homologues in Mycobacteria
Authors: Kaneda Kenji , Imaizumi Sadao , Yano Ikuya 
Published: November 2013

AbstractThere are three mycolic acid homologues with C22‐, C24‐ and C26‐α‐units in Mycobacterium. In order to reveal the composition and distribution of these homologues in each subclass and molecular species of mycolic acids and to compare them with the composition of constitutive non‐polar fatty acids (free and bound forms), we have separated non‐polar fatty acids and each subclass of mycolic acids from 21 mycobacterial species by thin‐layer chromatography, and analyzed non‐polar fatty acid methyl esters by gas chromatography (GC) and the cleavage products of methyl mycolate by pyrolysis GC. We further performed mass chromatographic analysis of trimethylsilyl (TMS) ether derivatives of mycolic acid methyl esters by monitoring [B‐29]+ ions (loss of CHO from the α‐branched‐chain structure of mycolic acids) of m/z 426, 454 and 482 which are attributed to C22‐, C24‐ and C26‐α‐units of TMS ether derivatives of methyl mycolates, respectively, (Kaneda, K. et al, J. Clin. Microbiol. 24: 1060‐1070, 1986). By pyrolysis GC, C22:0, C24:0 and C26:0 fatty acid methyl esters generated by the C2‐C3 cleavage of C22‐, C24‐ and C26‐α‐unit‐containing mycolic acid methyl esters, respectively, were detected. Their proportion was almost the same among subclasses of mycolic acids in every Mycobacterium and also similar to the proportion of constitutive non‐polar C22:0, C24:0 and C26:0 fatty acids. By mass chromatography, the composition and distribution of C22‐ and C24‐α‐unit‐containing homologues were revealed to be similar between α‐ and α'‐mycolic acids in every Mycobacterium. We further analyzed in detail M. vaccae and demonstrated that the mass chromatogram of C22‐α‐unit‐containing homologue was analogous in shape to that of the C24‐α‐unit‐containing one, with the latter mass chromatogram being up‐shifted from the former by two carbon numbers, in every subclass of α‐, α'‐, keto and dicarboxy mycolic acids. The present study suggests that the compositions of three homologues of both mycolic acids and constitutive non‐polar fatty acids, which are characteristic to each mycobacterial species, may reflect the proportion of the amount of free C22:0, C24:0 and C26:0 fatty acids synthesized in the cell. It is further demonstrated that intermolecular condensation of two fatty acids which become α‐ and β‐units of mycolic acids will occur independently of the carbon chain length or kinds of polar moieties of fatty acids.
Seasonal Distribution of Adenoviruses, Enteroviruses and Reoviruses in Urban River Water
Authors: Tani Naoto , Dohi Yoshiko , Kurumatani Norio , Yonemasu Kunio 
Published: November 2013

AbstractIn the 63‐month period from January 1988 to March 1993, monthly levels of adenoviruses, enteroviruses (coxsackie B, polio, echo) and reoviruses in the urban river water in Nara Prefecture, Japan were in the range 0‐25, 0‐190 and 0‐325, plaque forming units per liter (PFU/liter), and the average levels were 2.4, 40.6 and 56.2 PFU/liter, respectively. The peak reovirus level was found in winter during the cold weather months (Nov. to Mar.). The peak enterovirus level was found in summer (May to Sept.) but continued to be found in autumn‐winter (Oct. to Jan.) from 1991 to 1993. The levels of adenoviruses were low throughout all 5 years, as compared to those of reoviruses and enteroviruses. Polioviruses were isolated following the administration of vaccine. Although a changing pattern of serotype prevalence was seen with the coxsackie B viruses and echoviruses from 1988 to 1993, this is not so for polioviruses, which remained almost unchanged for the five‐year period. Adenoviruses were isolated throughout all five years, though in small numbers. Reoviruses were isolated most frequently throughout five years.
Down‐Regulation of T‐Cell Proliferation in Response to Soluble Anti‐CD3 Antibodies through Development of Redirected Cytolytic Activity Eliminating Costimulatory Cells
Authors: Lwin Tint , Nakashima Izumi , Nagase Fumihiko 
Published: November 2013

AbstractCD4+ T‐depleted spleen cells (CD8+ T cells) activated by anti‐CD3 antibodies (aCD3) suppressed proliferation of CD8+ T‐depleted spleen cells (CD4+ T cells) and fresh normal T cells in response to aCD3. Antigen‐nonspecific cytolytic activity was induced in splenic CD8+ T cells by stimulation with aCD3 and showed the peak level on day 3, whereas cytolytic activity induced in CD4+ T cells was weak. Intact Ig but not F(ab')2 of aCD3 induced and mediated cytolytic activity. Correspondingly, the cytolytic activity induced by aCD3 was directed against target cells bearing Ig‐binding Fc‐receptor activity and cytolysis was inhibited by the addition of free Ig into the assay system. We showed that aCD3‐activated T cells carried a high level of aCD3 on their surface at the time after the peak proliferation when they attained high cytolytic activity. This raised the possibility that the anti‐CD3‐induced aCD3‐redirected cytolytic activity eliminated Fc‐receptor‐bearing costimulatory cells in the culture for down‐regulation of the T‐cell proliferation. This view was supported by partial restoration of anti‐CD3‐induced low responsiveness of CD8+ T cells by the addition of fresh costimulatory cells. These results suggested a new pathway of down‐regulation of T‐cell proliferation by aCD3‐activated cytolytic CD8+ T cells.
Auxiliary Method for Clonal Identification of Staphylococcus aureus by Protein Band Pattern of Released Proteins on SDS‐Polyacrylamide Gel
Authors: Seki Keiko , Sakurada Junji , Murai Miyo , Usui Akemi , Seong Hee Kyong , Jitsukawa Hiroko , Masuda Shogo 
Published: November 2013

AbstractA supportive method for clonal identification of Staphylococcus aureus strains was devised. Culture supernatant obtained by cellophane surface culture was subjected to sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) without performing any concentration procedure prior to electrophoresis. The combined use of cellophane surface culture and SDS‐PAGE was convenient for determining whether the strains belonged to the same clone or not when conducted in conjunction with other tests for bacteriological characterization.
Inhibition by Bacterial Lipopolysaccharide of Spontaneous and TNF‐α‐Induced Human Neutrophil Apoptosis In Vitro
Authors: Hachiya Osamu , Takeda Yuji , Miyata Hiroaki , Watanabe Hiroshi , Yamashita Takao , Sendo Fujiro 
Published: November 2013

AbstractIn the previous paper (Takeda et al, Int. Immunol., 5, 691‐694, 1993), we demonstrated that tumor necrosis factor‐α (TNF‐α) promptly accelerates apoptosis of human neutrophils in vitro. In order to determine the role of neutrophil apoptosis in defending against bacterial infection, we studied the effect of bacterial lipopolysaccharide (LPS) on this process. LPS inhibited spontaneous and TNF‐α‐induced human neutrophil apoptosis in vitro, as determined by 1) light and electron microscopy, 2) flow cytometry, and 3) agarose gel electrophoresis of DNA. Low concentrations of cycloheximide, a protein synthesis inhibitor, which alone did not affect neutrophil apoptosis, were able to reduce spontaneous apoptosis inhibition by LPS, suggesting the involvement of newly synthesized protein in this phenomenon.
Subcutaneous Growth of Staphylococcus aureus Concomitantly Inoculated with Ehrlich Ascites Tumor Cells
Authors: Murai Miyo , Seki Keiko , Sakurada Junji , Usui Akemi , Masuda Shogo 
Published: November 2013

AbstractIntratumoral growth of Staphylococcus aureus Cowan I‐derived AP332 was examined by subcutaneous inoculation of cocci in doses ranging from 18 to 1.8 × 105 CFU with Ehrlich ascites tumor cells. Inoculation of 18 CFU AP332 resulted in staphylococcal growth in one of five mice, and the proportion of mice established intratumoral infection increased with the initial inocula. Six other strains of S. aureus also grew in the tumor tissue, and none of the three strains of coagulase‐negative staphylococci grew at all. Ethanol‐killed tumor cells did not promote staphylococcal growth as vigorously as the live tumor cells, especially when the initial inoculum of AP332 was smaller than 104 CFU.
Identification of Influenza C Virus Phosphoproteins
Authors: Nishimura Hidekazu , Sugawara Kanetsu , Gao Peng , Muraki Yasushi , Hongo Seiji , Kitame Fumio , Nakamura Kiyoto 
Published: November 2013

AbstractThe HMV‐II cells infected with influenza C virus were labeled with inorganic [32P]phosphate to identify phosphorylated proteins. Analysis by radioimmunoprecipitation with antiviral serum or monoclonal antibodies revealed that three major structural proteins of the virus, hemagglutinin‐esterase (HE), nucleoprotein (NP), and matrix protein (M1) are all phosphorylated in both infected cells and virions. It was also observed that, in the presence of trypsin (10 μg/ml), the unphosphorylated form of the HE glycoprotein was cleaved efficiently whereas the phosphorylated form was not, raising the possibility that phosphorylation of HE may influence its susceptibility to degradation by proteolytic enzymes.
Sequence Analysis of Flanking Regions of the pfoA Gene of Clostridium perfringens: β‐Galactosidase Gene (pbg) Is Located in the 3′‐Flanking Region
Authors: Shimizu Tohru , Kobayashi Takumi , Ba‐Thein William , Ohtani Kaori , Hayashi Hideo 
Published: November 2013

AbstractThe 3′‐flanking region of the perfringolysin O (theta‐toxin) gene (pfoA) of Clostridium perfringens was analyzed by chromosome walking. A total of 5,363 bp of the downstream region of the pfoA gene was sequenced and four open reading frames were found. ORF54 and ORF80 were found to be homologous to genes coding for membrane‐bound transporter proteins of other bacteria and the β‐galactosidase gene (bgaB) of Bacillus stearothermophilus, respectively. ORF80 was named the pbg gene. Clones which showed β‐galactosidase activities were selected from a λFIXII genomic library of C. perfringens by blue plaque screening using X‐Gal as a substrate. Four clones whose plaques showed blue appearances were obtained. Two of the four clones hybridized with the pbg probe but the others did not, indicating that there are two distinct β‐galactosidase genes in C. perfringens. The pbg gene was subcloned into pBR322 and was successfully expressed in Escherichia coli, suggesting that the pbg gene codes for a β‐galactosidase of C. perfringens.
Identification of the Sites for Suppressor Mutations on the Hemagglutinin Molecule to Temperature‐Sensitive Phenotype of the Influenza Virus
Authors: Tong Ning , Nakajima Katsuhisa , Nakajima Setsuko 
Published: November 2013

AbstractA temperature‐sensitive (ts) mutant of the influenza virus A/WSN/ 33 strain, ts‐134, possessed a defect in intracellular transport at the nonpermissive temperature and marked thermolability of hemagglutinin (HA) activity at 51 C. These were caused by a change at amino acid residue 157 from tyrosine to histidine in the HA protein. We isolated 37 spontaneous revertant clones from ts‐134 at the nonpermissive temperature and determined their HA sequences. The deduced amino acid sequences demonstrated that one was a true revertant and the others were revertants with suppressor mutations, each of which had an additional amino acid change besides those of ts‐134. The changed amino acids were located at 14 positions on the HA molecule, and eight of them were found in multiple revertants. These were located in five to six distinct regions on the three‐dimensional structure of the HA molecule. However, the heat stability of HAs in the revertants was recovered differently depending on the sites of the changed amino acids. The kinetics of transport of the HA protein in the revertants were slightly delayed compared to the wild‐type both at permissive and nonpermissive temperatures.
Detection of Astroviruses from Stool Samples in Japan Using Reverse Transcription and Polymerase Chain Reaction Amplification
Authors: Saito Kunihiro , Ushijima Hiroshi , Nishio Osamu , Oseto Mituaki , Motohiro Hiroshi , Ueda Yuichi , Takagi Michio , Nakaya Shigekazu , Ando Tamie , Glass Rodger , Zaiman Kohji 
Published: November 2013

AbstractWe developed a reverse transcription and polymerase chain reaction (RT‐PCR) method for detecting astrovirus serotypes 1, 2, 3, 5, 6 and 7 (but not serotype 4). Furthermore, we developed the specific primers for detecting serotypes 1 and 2, the most predominant serotypes in the world. Sensitivity of the first PCR with serotype common primers was about 10 times higher than that of enzyme immunoassay with monoclonal antibody (EIA‐MAb). Sensitivity of the second PCR with the serotype‐specific primers was even higher. The RT‐PCR method was useful for detecting astroviruses from clinical samples, especially serotypes 1 and 2.
Genetic Variation in VP7 Gene of Human Rotavirus Serotype 2 (G2 Type) Isolated in Japan, China, and Pakistan
Authors: Wen Leying , Ushijima Hiroshi , Kakizawa Junko , Fang Zhao‐Yin , Nishio Osamu , Morikawa Shigeru , Motohiro Takashi 
Published: November 2013

AbstractSequence analysis of the gene encoding the major neutralization glycoprotein (VP7) was performed on sixteen human isolates of serotype 2 of rotavirus in Japan, China, and Pakistan and their genetic variations were examined. Comparative studies of their nucleotide and deduced amino acid sequences between the sixteen isolates and the HU5 strain revealed an overall homology of more than 94%. A higher degree of homology in nucleotides was observed among the sixteen isolates than between HU5 and the isolates. A total of thirteen amino acid residues frequently converted to another amino acid. Out of the thirteen, five amino acid residues belonging to the major neutralizing epitope regions (C, E, and F in this communication) converted frequently. From the amino acid sequences three subtypes, subtype 1, subtype 2, and intermediate, were suggested to be classified as previously reported for serotype 1 (Xin et al, Virology, 1993, 197: 813‐816).
Major Enteropathogenic Bacteria Isolated from Diarrheal Patients in Bolivia: A Hospital‐Based Study
Authors: Utsunomiya Akiyoshi , Elío‐Calvo Daniel , Reyes Armando Alberto Benitez , Castro Ernesto Sanzetenea , Rodríguez Enrique , Tress Carolina , de Corzo Jenny I. Zamora , Hannover Erika , Kai Akemi , Tamura Kazumichi , Higa Naomi 
Published: November 2013

AbstractA total of 1,234 fecal samples from diarrhea cases were examined for etiological bacterial agents at medical facilities in La Paz and Sucre, Bolivia. Eighty strains of Shigella spp., 39 strains of Salmonella spp., 29 strains of Vibrio cholerae, and 222 strains of enteropathogenic Escherichia coli (139 EPEC, 55 ETEC, 29 EIEC, and 1 EHEC) were isolated. With regard to the serovars of Shigella, S. flexneri 2a, 3a, and 1b were predominant. In the case of Salmonella, S. enteritidis was the most common, followed by S. typhi, S. poona, and S. paratyphi B. Out of 29 cholera strains, 25 belonged to biovar El Tor, serovar Ogawa while the remaining 4 were serovar Inaba. Among 55 strains of ETEC serotypes, 5 showed ST producers but none showed LT producers. Likewise, among 55 strains of enterohemorrhagic serotypes, only one strain (O157:H7) produced verocytotoxin (VT 2). The results of drug sensitivity tests revealed the predominance of Shigella, EPEC, and ETEC strains resistant to aminobenzil‐penicillin (ABPC) and trimethoprim. Since diarrheal patients in Bolivia are treated mainly with ABPC or sulfamethoxazole/trimethoprim (SXT) and rarely with gentamicin, kanamycin, or other drugs, it is possible that ABPC‐ and SXT‐resistant strains will increase and persist in the near future.
Sequence Analysis of the Streptococcus mutans Ingbritt dexA Gene Encoding Extracellular Dextranase
Authors: Igarashi Takeshi , Yamamoto Ayako , Goto Nobuichi 
Published: November 2013

AbstractThe complete nucleotide sequence (3,747 bp) of the dextranase gene (dexA) and flanking regions of the chromosome of Streptococcus mutans Ingbritt (serotype c) were determined. The open reading frame for dexA was 2,550 bp, ending with a stop codon TGA. A putative ribosome‐binding site, promoter preceding the start codon, and potential stem‐loop structure were identified. The presumed dextranase protein (DexA) consisting of 850 amino acids was estimated to have a molecular size of 94,536 Da and a pI of 4.79. The nucleotide sequence and the deduced amino acid sequences of S. mutans dexA exhibited homologies of 57.8% and 47.0%, respectively, to those of Streptococcus sobrinus dex. The homologous region of dex of S. sobrinus was in the N‐terminal half. The C terminus of DexA consisted of a hexapeptide LPQTGD, followed by 7 charged amino acids, 21 amino acids with a strongly hydrophobic character, and a charged hexapeptide tail, which have been reported as a common structure of C termini of not only the surface‐associated proteins of Gram‐positive cocci but also the extracellular enzymes such as β‐fructosidase of S. mutans and dextranase of S. sobrinus. The DexA protein had no significant homology with the glucosyltransferases, the glucan‐binding protein, or the dextranase inhibitor of mutans streptococci.
Expression of 18.6/CD23 Antigen on Human Lymphoid Progenitor Cell Lines and Phorbol 12‐Myristate 13‐Acetate (PMA)‐Induced Microglia‐Shaped Cells
Authors: Okumura Akiou , Kishi Hiroyuki , Tagoh Hiromi , Minowada Jun , Muraguchi Atsushi 
Published: November 2013

AbstractThe nature of lymphoid progenitors and factor(s) determining commitment to either the T‐ or B‐lymphocyte pathway are poorly understood in the human system. In this study, we generated a monoclonal antibody (MoAb), 18.6, that recognizes a cell surface antigen on a human lymphoid progenitor cell line (FL4.4). MoAb 18.6 reacted with lymphoid progenitor lines, B lymphoid cell lines, and myelomonocytic cell lines. It did not react with any T cell or erythroid leukemic cell lines. Two color FACS analyses of normal lymphoid tissues showed that MoAb 18.6 reacted with a majority of CD20+ mature B cells and a minority of CD64+ monocytes. Molecules of 3 different sizes with MW of 34, 45, and 68 Kd were precipitated with MoAb 18.6 from the lymphoid progenitor cell line. The 18.6 antigen was not expressed on a fetal liver‐derived lymphoid progenitor‐like cell line, FL1.4, which has the capacity to differentiate into microglia‐shaped cells upon PMA‐stimulation. Stimulation of FL1.4 cells with PMA induced expression of the 18.6 antigen within 24 hr and the microglia‐shaped cells stained positively with MoAb 18.6. Finally, cloning of a cDNA that encoded the 18.6 antigen revealed that the 18.6 antigen is identical to the CD23 antigen. Taken together, these data suggest that the 18.6/CD23 antigen is expressed on lymphoid precursors at a very early stage of differentiation.
Prevention of Vascular Graft Infection by Sisomicin Incorporated into Fibrin Glue
Authors: Yamamura Keiko , Sakurai Tsunehisa , Yano Kohji , Osada Takashi , Nabeshima Toshitaka 
Published: November 2013

AbstractTo examine the efficacy of sisomicin (SISO) incorporated into fibrin glue (FG) for the prevention of graft infection in animal models, the susceptibility to infection of Dacron grafts (control) and SISO‐FG Dacron grafts following the inoculation of Staphylococcus aureus or S. epidermidis was compared. The results showed that SISO‐FG Dacron grafts displayed resistance to graft infection.
Correlation of the Polysaccharide Antigens of Francisella tularensis with Virulence in Experimental Mice
Authors: Fujita Hiromi , Sato Tadashi , Watanabe Yuriko , Ohara Yoshiro , Homma Morio 
Published: November 2013

AbstractFrancisella tularensis gives rise to two distinct colony types, acriflavine agglutination test‐positive (acf+) and ‐negative (acf−) colonies. The acf+ variants were exclusively low virulent in mice, while the acf− variants were shown to be either high or low virulent. Three fractions, phosphate‐buffered saline‐extractable without heating, with heating at 60 C, and with heating at 100 C, were obtained from cultures of both the acf+ and acf− variants on agar media, and the polysaccharide antigens in those fractions were quantitated. All of the highly virulent acf− variants possessed a large amount of the polysaccharide antigen in the fraction extractable with heating at 60 C. This antigen was not, however, detected in any of the acf+ variants and one low‐virulent acf− variant. It was also detected in a very low amount in some other acf− variants with low virulence. The amount of this polysaccharide antigen was therefore shown to be correlated with bacterial virulence in mice.
T Cell Response to Borrelia garinii, Borrelia afzelii, and Borrelia japonica in Various Congenic Mouse Strains
Authors: Ishii Norihisa , Isogai Emiko , Isogai Hiroshi , Kimura Koichi , Nishikawa Takeshi , Fujii Nobuhiro , Nakajima Hiroshi 
Published: November 2013

AbstractThe infectivity and T cell response to Borrelia garinii SIKA2, Borrelia afzelii BFOX, and Borrelia japonica 0612, the organisms that cause Lyme disease in Japan, were examined in various inbred and congenic strains of mice. Infectivity differed among the species: B. garinii SIKA2 and B. afzelii BFOX were each able to infect 90% to 100% of C3H/He mice; B. japonica 0612 was able to infect only 20% of C3H/He mice. The pattern of infectivity to various inbred and congenic strains of mice may influence the pathogenicity of the organism and the clinical signs of Lyme disease. Cross‐reactivity between Borrelia antigens was observed, but there was no cross‐reactivity between Borrelia antigens and Leptospira antigens. We evaluated the genetic control of the delayed‐type hypersensitivity (DTH) reaction in the form of footpad swelling produced by Borrelia antigens using viable or sonicated bacteria as sensitization. Differences in strains of mice infected by viable antigen were observed. However, all strains of mice showed a strong DTH reaction using sonicated antigens without genetic background. A DTH reaction in the form of footpad swelling did not appear to be associated with genetic background. The footpad reaction was mediated by CD4+8− and Ia− T cells, as revealed by in vitro monoclonal antibody treatment. However, CD8+ T cells did not suppress footpad swelling. These results indicate that many antigenic epitopes of the Borrelia spirochete can stimulate the DTH reaction.