bluuebunny/crossref_metadata_2025_split huggingface dataset:
Crystallization and Analyses of Crystals of Various Chemotypes of R‐Form Lipopolysaccharides from Salmonella Spp.
Authors: Kato Nobuo , Arakawa Yoshichika , Sugiyama Tsuyoshi , Ito Hideo , Naito Setsuko , Kido Nobuo , Ohta Michio , Sasaki Kyoyu 
Published: November 2013

AbstractVarious chemotypes (Re, Rd2, Rd1P–, Rd1, RcP–, Rc, Rb3, Rb2, Rb1, and Ra) of R‐form lipopolysaccharides (LPSs) of Salmonella spp. were crystallized by treatment with 70% ethanol containing 250 mM MgCl2, and crystals of the LPSs were observed electron microscopically and analyzed by electron diffraction and synchrotron X‐ray diffraction. All the LPSs tested formed three‐dimensional crystals showing very similar shapes; hexagonal plate, solid column, discoid, square or rectangular plate, lozenge plate and truncated hexangular or rectangular pyramid forms. Electron diffraction patterns from the hexagonal plate crystals of all these LPSs obtained by electron irradiation from the direction perpendicular to the basal plane showed that they consist of hexagonal lattices with the lattice constant of 4.62 Å. The crystals of all the LPSs thus formed gave ring‐like X‐ray diffraction patterns because of their small sizes. The long‐axis values were calculated from the X‐ray diffraction patterns from crystals of all the LPSs in the low‐angle region and they corresponded roughly to the length of the proposed primary chemical structures of the R cores of the LPSs. The volume occupied by a single molecule of all the LPSs were calculated from the molecular weights based on the proposed structures and the crystallographic data obtained by electron diffraction, X‐ray diffraction, and density determination.
Siderophore‐Mediated Utilization of Iron Bound to Transferrin by Vibrio parahaemolyticus
Authors: Yamamoto Shigeo , Okujo Noriyuki , Matsuura Shyoko , Fujiwara Ikuko , Fujita Yasuhiro , Shinoda Sumio 
Published: November 2013

AbstractVibrio parahaemolyticus produces a structurally novel type of siderophore, termed vibrioferrin, in response to iron‐limitation. This study was performed to examine whether vibrioferrin can assimilate iron from human iron‐binding proteins for growth. Comparison of the growth rates between V. parahaemolyticus AQ 3354 and its spontaneously arising, vibrioferrin‐deficient mutant revealed that vibrioferrin was able to sequester iron from 30% iron‐saturated human transferrin for growth, but not from human lactoferrin even if fully saturated with iron. In both strains, iron limitation induced two high‐molecular‐weight outer membrane proteins with apparent molecular masses of approximately 78 and 83 kDa. Since only the outer membrane fraction including these proteins showed a binding capacity to ferric vibrioferrin complex, either of them may function as its cell surface receptor. These results suggested that the organism might utilize such a source of host iron through the action of vibrioferrin during in vivo survival and proliferation, although its importance in pathogenesis is unknown.
Pulsed Field Gel Electrophoresis Analysis of Borrelia burgdorferi Sensu Lato Isolated in Japan and Taxonomic Implications with Lyme Disease Spirochetes
Authors: Fukunaga Masahito , Takahashi Yukie 
Published: November 2013

AbstractGenomic DNAs of Borrelia burgdorferi sensu lato isolates obtained in Japan sharing different rRNA gene ribotypes were digested with rare‐cutter restriction endonucleases and the fragments obtained were separated by pulsed field gel electrophoresis (PFGE). The sizes of large restriction cleavage bands with MluI endonuclease were quite similar among isolates in each ribotype group. On the other hand, the PFGE profiles obtained with the other enzymes (NruI, Sal I or SplI) were rather divergent, and Japanese isolates were distinguishable from the United States and European isolates. The Japanese isolates classified as ribotypes group II (Borrelia garinii) and III (B. afzelii) showed different PFGE patterns from that of European isolates. The isolates grouped into ribotype IV revealed distinctively different PFGE profiles. These results indicate that the Japanese isolates may be genetically divergent and distinct from the United States and European isolates.
Complement‐Mediated Killing of Borrelia garinii —Bactericidal Activity of Wild Deer Serum
Authors: Isogai Emiko , Kamewaka Yuri , Isogai Hiroshi , Kimura Koichi , Fujii Nobuhiro , Nishikawa Takeshi 
Published: November 2013

AbstractThe susceptibility of Borrelia garinii to fresh wild deer sera was determined by incubating strain SIKA2 at 10% serum concentration for 1 hr at 37 C in an in vitro bactericidal assay. Each serum showed bactericidal effects at various levels. The effect was dependent on the concentration of antibody to the spirochetes. Complement was essential in the bactericidal assay because the inactivated deer serum showed greatly decreased activity. Our results suggest that B. garinii is sensitive to deer serum, in the presence of antibody and the bactericidal effect is important for preventing Lyme disease in wild sika deer.
Isolation and Classification of Temperature‐Sensitive Mutants of Influenza B Virus
Authors: Yamamoto‐Goshima Fumi , Aoki Hiizu , Takada Toshihiko , Toda Shigeru , Yoshii Saiji , Maeno Koichiro 
Published: November 2013

AbstractWe isolated 25 temperature‐sensitive mutants of B/Kanagawa/73 strain generated by mutagenesis with 5‐fluorouracil and classified them into seven recombination groups by pair‐wise crosses. All mutants showed a ratio of plaquing efficiency at the nonpermissive temperature (37.5 C) to the permissive temperature (32 C) of 10–4 or less. At 37.5 C most of group I, II, and III mutants did not produce appreciable amounts of protein, but all other group mutants were protein synthesis‐positive. A group VII mutant produced active hemagglutinin (HA) and neuraminidase (NA) at the nonpermissive temperature, but Group V mutants produced only active NA and were defective in the HA molecule. The other group mutants, including group IV mutants with mutation only in the NA gene (8, 10), lacked both activities at the nonpermissive temperature. One of nine influenza B virus isolates in 1989 had EOP 37.5/32 of 1/3 × 10–2 and belonged to recombination group VII.
Expression of Fimbriae and Host Response in Branhamella catarrhalis Respiratory Infections
Authors: Ahmed Kamruddin , Masaki Hironori , Dai Iran Cong , Ichinose Akitoyo , Utsunomiya Yoshiaki , Tao Misao , Nagatake Tsuyoshi , Matsumoto Keizo 
Published: November 2013

AbstractSputum during the acute exacerbation of chronic respiratory diseases were observed under the electron microscope, to determine the in vivo expression of surface structures of Branhamella catarrhalis (B. catarrhalis), the polymorphonuclear neutrophil (PMN) response to B. catarrhalis infections, and the composition of sputum. It was found that during infection fimbriae are expressed in B. catarrhalis. However, there were sparsely to densely fimbriated bacteria in each sputum sample. The length of the fimbriae were from 50 to 76 nm. In the sparsely fimbriated B. catarrhalis, external to the cell wall, a thin, granular, electron‐dense layer was observed. Due to the presence of fimbriae, this layer was not seen in densely fimbriated B. catarrhalis. Blebs were also found in B. catarrhalis. PMNs were found to phagocytose both B. catarrhalis and debris. Evidence was found that debris were formed mainly by the destruction of PMNs. Bacteria as well as debris were phagocytosed by PMNs.
Use of the Recombinant 38‐kDa Antigen of Mycobacterium tuberculosis as an Immunogen for Specific Antisera Production
Authors: D'Souza Celine D. , Kadival Gururaj V. , Samuel Aban M. 
Published: November 2013

AbstractThe Mycobacterium tuberculosis 38‐kDa protein antigen is one of the secreted immunodominant antigens showing high immunogenicity at B‐cell and T‐cell levels. Although monoclonal antibodies to this antigen have been produced, specific polyclonal antisera is required for standardization of specific immunodiagnostic assays. This protein has been overexpressed and purified from recombinant Escherichia coli using an inducible vector system. During each stage of expression and purification, the recombinant protein was used to immunize mice and rabbits by several methods: 1) as overexpressed protein present as inclusion bodies in recombinant E. coli; 2) embedded in a polyacrylamide gel; 3) fixed to a solid‐phase nitrocellulose membrane and 4) emulsified with an adjuvant. All strategies yielded specific antisera as determined by enzyme‐linked immunosorbent assay (ELISA) and immunoblot analyses. The results obtained, both quantitative (ELISA) and qualitative (immunoblot) demonstrate that the purified recombinant antigen retains its antigenicity and immunogenicity throughout the various steps in the process of expression and purification and serves as a potent antigen for production of specific antisera to be used in immunoassays.
Borrelia japonica in Nature: Genotypic Identification of Spirochetes Isolated from Japanese Small Mammals
Authors: Nakao Minoru , Miyamoto Kenji , Fukunaga Masahito 
Published: November 2013

AbstractSpirochetes were isolated from earlobe tissues of shrews (Sorex unguiculatus, Sorex caecutiens, and Crocidura dsinezumi), voles (Clethrionomys rufocanus), and mice (Apodemus argenteus and Apodemus speciosus) captured in various localities in Japan. The isolates were identified as Borrelia japonica by rRNA gene restriction fragment length polymorphism analysis. The data suggest that these small mammals are candidates of reservoir hosts for B. japonica.
Comparative Analyses of Thymocyte and Thymic Low‐Density Adherent Cell Functions
Authors: Wambua Peter P. , Iwabuchi Kazuya , Iwabuchi Chikako , Ogasawara Kazumasa , Itoh Yasushi , Arase Hisashi , Kajiwara Masaharu , Gotohda Toshihiko , Kajino Ki‐ichi , Good Robert A. , Onoé Kazunori 
Published: November 2013

AbstractThymocytes which have developed in the C3H thymus showed depressed proliferative responses to stimulation with anti‐CD3 antibody as compared with those which have developed in the thymus of other strains of mice (i.e. AKR). The present study was conducted to analyze immunological functions of the thymic stromal cell population (low‐density adherent cells, LDAC) in the C3H mice using allogeneic bone marrow (BM) chimeras established by BM transplantation in the reciprocal combination of AKR and C3H mice as donor or recipient. The thymic LDAC from C3H mice or the [AKR(donor)→C3H(recipient)] chimeras contained a high proportion of Mac‐1+ cells as compared to AKR mice or the [C3H→AKR] chimeras. The proportion of Mac‐1+ cells paralleled the IL‐1‐ and PGE2‐secreting ability of the LDAC cultured either in the presence or absence of LPS and also paralleled the antigen‐presenting cell functions of the LDAC. Furthermore, after anti‐CD3 stimulation the PGE2 inhibited more profoundly proliferative responses of [AKR→C3H] or normal C3H thymocytes than those of the [C3H→AKR] chimera or normal AKR thymocytes. A PGE2 inhibitor, indomethacin, reversed the depressed responses of the thymocytes which had developed in the C3H thymus. These findings suggest that the lower responsiveness of thymocytes from [AKR→C3H] chimeras to anti‐CD3 stimulation may be attributable to large amounts of PGE2 secreted by LDAC and/or to increased sensitivity of thymocytes themselves to PGE2.
Increased Phosphodiesters in Lipopolysaccharide Prepared from the Polymyxin B‐Resistant Isolate of Klebsiella pneumoniae
Authors: Yokochi Takashi , Inoue Yoshiko , Jiang Guo‐zhi , Kato Yutaka , Sugiyama Tsuyoshi , Kawai Makoto , Fukada Masako , Takahashi Kazuko 
Published: November 2013

AbstractA polymyxin B (PXB)‐resistant mutant of Klebsiella pneumoniae O3 was isolated. Lipopolysaccharide (LPS) extracted from the PXB‐resistant isolate bound little PXB, although LPS from the parental strain did. The 31P nuclear magnetic resonance (NMR) spectrum of PXB‐resistant type LPS showed that it contained much less of the phosphomonoesters and the pyrophosphate esters, and an increased amount of the phosphodiesters, compared to the parental type LPS. The decrease in the binding of PXB might be due to altered phosphate groups on the PXB‐resistant type LPS, suggesting that it might explain the PXB‐resistance of the mutant.
Isolation and Characterization of Three Porin‐Like Proteins from Vibrio vulnificus: Effect of Different Growth Media on Their Production
Authors: Koga Tetsuro , Takumi Kenji 
Published: November 2013

AbstractThe effect of the culture media on the composition of the outer membrane protein of Vibrio vulnificus strain 393 from human blood was examined. Only one major outer membrane protein, with an apparent molecular weight of 37,000 (37K protein) and 34,000 (34K protein), was formed in the cells grown in 3% NaCl‐BHI broth and chemically defined medium, respectively. The production of one major outer membrane protein was also observed in other isolates from humans and asari clam when they were grown in 3% NaCl‐BHI broth. On the other hand, three major outer membrane proteins, with apparent molecular weights of 48,000 (48K protein), 37,000 (37K protein), and 34,000 (34K protein), were produced in the cells grown in 3% NaCl‐nutrient broth. Three proteins, 48K, 37K, and 34K from strain 393, were purified and the amino acid compositions were determined. Although there was a little difference in the composition of amino acid among three proteins, the amino acid compositions of the three porin‐like proteins showed characteristic properties of the porins of Escherichia coli and Salmonella typhimurium. Immunoblot analysis of the outer membrane proteins from four vibrios, E. coli, and S. typhimurium using monospecific antisera against these three porin‐like proteins showed that only the antiserum against 37K protein cross‐reacted with the outer membrane proteins from all the strains tested.
Suppression of Anti‐Candida Activity of Murine and Human Neutrophils by Glucocorticoids
Authors: Nohmi Takao , Abe Shigeru , Tansho Shigeru , Yamaguchi Hideyo 
Published: November 2013

AbstractEffects of glucocorticoid (GC) compounds on inhibitory activity of neutrophils to mycelial growth of Candida albicans were examined by in vitro crystal violet staining method with 14 hr co‐culture. Both GC hormones (hydrocortisone ≥6 × 10–7 m and corticosterone ≥10–6 m) and anti‐inflammatory GC agents (prednisolone ≥10–7 m and dexamethasone ≥10–8 m) significantly suppressed anti‐Candida activity of murine casein‐induced neutrophils. Anti‐Candida activity of human neutrophils prepared from peripheral blood was also suppressed by hydrocortisone (≥6 × 10–7 m). These GC compounds did not affect the Candida growth in the absence of neutrophils. Steroidal compounds without anti‐inflammatory activity, cholesterol, cholic acid, aldosterone did not suppress neutrophil activity. These results suggest that GCs at their physiological or clinical concentration may suppress anti‐Candida activity of neutrophils in vivo.
Isolation and Characterization of Staphylococcus aureus Mutants Which Form Altered Cell Clusters
Authors: Sugar Motoyuki , Komatsuzawa Hitoshi , Ooku‐Inomata Kaoru , Miyake Yoichiro , Ishida Eisaku , Suginaka Hidekazu 
Published: November 2013

AbstractStaphylococcus aureus FDA 209P produces two extracellular bacteriolytic enzymes, 51‐kDa endo‐β‐N‐acetylglucosaminidase (GL) and 62‐kDa N‐acetylmuramyl‐l‐alanine amidase (AM), both of which can disperse cell clusters. To characterize the physiological roles of these enzymes in vivo, mutants with altered autolysin activity were isolated, and their degree of cluster formation in broth culture was assessed. Bacteriolytic activities of GL and AM, produced and secreted from these mutants into the culture fluid and detected with activity gels, coincided well with the degree of cluster formation of the mutants. The mutants with little or no enzyme activity grew in clusters, whereas those with high activity grew as well‐separated cocci, suggesting that these enzymes are involved in cell separation of S. aureus in vivo.
Seroepidemiological Survey of Coxiella burnetii in Domestic Cats in Japan
Authors: Morita Chiharu , Katsuyama Jun , Yanase Tsuyoshi , Ueno Hiroshi , Muramatsu Yasukazu , Hohdatsu Tsutomu , Koyama Hiroyuki 
Published: November 2013

AbstractCats are assumed to be one of the most important reservoirs of causative agent of human Q fever especially in urban areas. There is no evidence of Coxiella burnetii infection in cats in Japan prior to this. Sera from 100 cats, collected in various parts of Japan, were examined for antibody against C. burnetii. Sixteen out of the 100 samples contained antibodies against C. burnetii. The prevalence of the antibody decreased from the northeastern to the southwestern part of Japan. A high prevalence of the antibodies was observed in sera from cats of more than four years of age. It is difficult to deny that cats would be one of the important sources of human Q fever in Japan.
Purification and Characterization of Metallo‐β‐Lactamase from Serratia marcescens
Authors: Marumo Kenji , Takeda Atsushi , Nakamura Yoshiko , Nakaya Kazuyasu 
Published: November 2013

AbstractCarbapenem‐hydrolyzing β‐lactamase from Serratia marcescens FHSM4055 was purified 926‐fold by means of carboxylmethyl Sephadex C‐50, Sephacryl S‐200, and Mono S column chromatography. The molecular weight was 30,000 by SDS‐PAGE and the isoelectric point was 8.7. The enzyme activity was inhibited by EDTA, and restored by adding zinc (II) or manganese (II). It was inhibited by p‐chloromercuribenzoate and iodine as well as the heavy metals, Hg (II), Fe (II), Fe (III), and Cu (II). These results indicate that the enzyme is a metallo‐β‐lactamase and that the SH‐group of only one cysteine residue probably binds to the metal ion, thus contributing to the stability of the enzyme active center. The specific constant (kcat/Km) showed that the enzyme hydrolyzed various β‐lactam antibiotics such as carbapenems, cephalosporins, moxalactam, cephamycins, and penicillins other than monobactams. Ampicillin and piperacillin with respective amino‐ and imino‐groups, ceftazidime with a carboxypropyloxyimino‐group, and cefclidin with a carbamoylquinuclidine‐group were poor substrates among the β‐lactam antibiotics other than the monobactams tested. The plots of the turnover number (kcat) against pH for the hydrolysis of cephaloridine gave an asymmetrical curve with the ‘tail’ on the acid side (pK1, 5.9; pK2, 9.0; pK3, 10.8), whereas those of kcat/Km gave a bell‐shaped curve (pK1, 5.8; pK2, 9.8). Both results suggest that two ionic forms of an intermediate yield the same product at different rates and that the enzyme is stable under alkaline conditions. Since the N‐terminal amino acid sequence of 27 residues determined was consistent with that of the metalloenzyme (Antimicrob. Agents Chemother., 1994, 38: 71‐78), the above enzymatic characteristics seem to coincide.
Analysis of Cellular Response and Gamma Interferon Synthesis in Bronchoalveolar Lavage Fluid and Lung Homogenate of Mice Infected with Pneumocystis carinii
Authors: Ishimine Tomohiko , Kawakami Kazuyoshi , Nakamoto Atsushi , Saito Atsushi 
Published: November 2013

AbstractThe cellular and cytokine responses in the lungs of mice infected with Pneumocystis carinii were examined on both lung homogenates and bronchoalveolar lavage (BAL) fluids. In the lungs of infected mice, the number of P. carinii cysts rapidly decreased by day 7, then started to increase with a peak on day 14, and thereafter decreased gradually. When the presence of P. carinii was examined at the DNA level by dot blot hybridization, a similar clearance curve was obtained, and the organisms were shown to be completely eliminated on day 28. In the late phase of infection, leukocytes, mainly lymphocytes, increased in number when analyzed on lung homogenates, while no significant increase of inflammatory cells was observed in BAL fluids. An accumulation of both CD4+ and CD8+ T cells and an increase of activated T cells expressing IL‐2Rα were observed in lung homogenates of the infected mice. In addition, a considerable amount of IFN‐γ was detected in lung homogenates, but not in BAL fluids. These data indicate that lung homogenates are more suitable than BAL fluids for the analysis of cellular and cytokine responses in the lungs of mice infected with P. carinii. To define the involvement of IFN‐γ in host defense against P. carinii, the effect of this cytokine on the killing activity of macrophages against P. carinii was examined in vitro. IFN‐γ was found to augment this activity by increasing nitric oxide synthesis of the macrophages. Thus, it is suggested that IFN‐γ plays an important role in the protection of mice from P. carinii infection.
Production of Antigenically Related Exocellular Elastolytic Proteases Mediating Hemagglutination by Vibrios
Authors: Alam Munirul , Miyoshi Shin‐ichi , Shinoda Sumio 
Published: November 2013

AbstractExocellular proteases produced by Vibrio fluvialis, V. furnissii, V. metschnikovii and V. campbellii were characterized and compared to those of V. mimicus protease (VMP) and V. vulnificus protease (VVP). These proteases possessed both elastolytic and hemagglutinating abilities and were identified, except that of V. metschnikovii, as metalloprotease. Conversely, V. metschnikovii protease failed to exhibit some of the salient features for metalloproteases suggesting the existence of protease(s) other than metalloprotease. However, antibodies against VVP cross‐reacted to these proteases and to VMP indicating antigenic relatedness amongst vibrio proteases. This study, thus, demonstrated the prevalent distributions of antigenically related proteases both in pathogenic and non‐pathogenic vibrios, bringing their status as a virulence determinant into question.
Experimental Dental Caries on Gnotobiotic Inbred Mice
Authors: Maeda Nobuko , Arai Satoshi , Ozaki Akira , Oowada Tsutomu , Takahashi Akiko , Fujita Hiroshi , Mizutani Takeo 
Published: November 2013

AbstractThe purpose of this study in mono‐infected gnotobiotic BALB/cA and C3H/HeN mice was to evaluate the cariogenicity of Enterococcus faecalis. The caries incidence and mean caries score in the BALB/cA mice were significantly higher than those in the C3H/HeN. In both of the mouse strains, the mean number of E. faecalis isolated from the cecum content was almost the same, however, the mean number of E. faecalis from the maxilla of BALB/cA was significantly higher than that of C3H/HeN. These results indicate that C3H/HeN has some factors that prevent E. faecalis from attaching to the tooth surfaces.