bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of Cholera Toxin by a Highly Sensitive Bead‐Enzyme Linked Immunosorbent Assay
Authors: Uesaka Yoshihiko , Otsuka Yoko , Kashida Mitsuaki , Oku Yuichi , Horigome Kazuki , Nair G. Balakrish , Pal S. C. , Yamasaki Shinji , Takeda Yoshifumi 
Published: November 2013

AbstractA bead‐enzyme linked immunosorbent assay (bead‐ELISA) for detection and quantification of cholera toxin (CT) in broth cultures of Vibrio cholerae O1 has been developed. Under optimal buffer and pH conditions the bead‐ELISA could consistently detect 40 pg/ml of CT. None of the ingredients of commonly used media for in vitro culture of V. cholerae O1 hindered the performance of the bead‐ELISA. Evaluation of the sensitivity and specificity of the bead‐ELISA against the commonly used reversed passive latex agglutination (RPLA) test for detection of CT was performed using a collection of 239 strains of V. cholerae O1 (including both biotypes and serotypes) which were examined by a gene probe encoding for the A1 subunit of CT. Although both the assays were highly specific, the bead‐ELISA was more sensitive than the RPLA. Quantification of CT by the bead‐ELISA revealed that the concentration of CT produced by the strains of V. cholerae O1 which were negative by the RPLA was lower than 1 ng/ml and therefore below the minimum detection ability of the RPLA. The bead‐ELISA is a simple, specific and highly sensitive assay for routine detection of CT and is recommended for routine use in clinical microbiology laboratories.
Studies on Osmotic Stability of Liposomes Prepared with Bacterial Membrane Lipids by Carboxyfluorescein Release
Authors: Nagamachi Eiko , Hirai Yoshikazu , Tomochika Ken‐ichi , Kanemasa Yasuhiro 
Published: November 2013

AbstractThe authors measured the osmotic stability of liposomes prepared with membrane lipids of bacteria, using the osmotic‐shock release of entrapped carboxyfluorescein as an indicator. The sub‐second physical changes of liposomes suspended in a solution of low osmotic pressure were examined by stopped flow spectrophotometry. The entrapped carboxyfluorescein was released when the liposomes burst on inflow of excess water. Liposomes prepared with the lipids of a stable Staphylococcus aureus L‐form strain were more resistant to low osmotic pressure than those prepared from the wild strain of S. aureus, and liposomes prepared from Mycoplasma orale were even more resistant. Cardiolipin enhanced the lipid membrane stability in S. aureus and cholesterol in M. orale. The stability of lipid membranes to low osmotic pressure could be precisely determined by the present method.
Interferon Induction in Murine Peritoneal Macrophage by Stimulation with Lactobacillus acidophilus
Authors: Kitazawa Haruki , Matsumura Kazuki , Itoh Takatoshi , Yamaguchi Takahiro 
Published: November 2013

AbstractInduction of interferon for a kind of dairy lactic acid bacteria, Lactobacillus acidophilus (L. acidophilus), was investigated in murine peritoneal macrophage (MØ) cultures. Lactobacillus acidophilus JCM 1034, 1132T, 1229 and 2125 induced IFN (12–34 I.U./ml) in MØ cultures in vitro. Strain 1132T‐ and 2125‐induced IFNs were characterized as IFNα/β by treatment with anti‐IFNs serum. The results indicate that the inducing activity of IFNs may be one of the available biological parameters for designating the dairy products containing L. acidophilus as “physiologically functional foods.”
Lipopolysaccharide Composition and Virulence Properties of Clinical and Environmental Strains of Vibrio fluvialis and Vibrio mimicus
Authors: Rahman Mohammed Mahbubur , Qadri Firdausi , Albert Manuel John , Hossain Anwar , Mosihuzzaman M. 
Published: November 2013

AbstractVibrio mimicus strains W‐26768 (stool isolate) and N‐1301 (environmental isolate) and Vibrio fluvialis strains AA‐18239 (stool isolate) and M‐940 (environmental isolate) were studied for virulence properties and lipopolysaccharide composition. All four strains were hydrophobic, produced cytotoxin, adhered to HeLa cells and showed mannose‐sensitive agglutination of guinea pig erythrocyte. The strains were negative for enterotoxin production and were mostly susceptible to the common antibiotics. The environmental and clinical isolates of both species were anti‐genically unrelated to each other. Lipopolysaccharide antigen analysis showed that O‐antigen polysaccharides of two strains of V. fluvialis and two strains of V. mimicus differed with respect to the sugar components. Only LPS from V. mimicus W‐26768 showed the presence of an unusual sugar, 3,6‐dideoxy‐3‐acetamido‐hexose. The sugar compositions of these V. fluvialis and V. mimicus strains differed from those of previously reported Japanese isolates. These differences probably reflect differences in the serogroup of strains.
Bacteriostatic and Bactericidal Activity of Antituberculosis Drugs against Mycobacterium tuberculosis, Mycobacterium avium‐Mycobacterium intracellulare Complex and Mycobacterium kansasii in Different Growth Phases
Authors: Yamori Sadaaki , Ichiyama Satoshi , Shimokata Kaoru , Tsukamura Michio 
Published: November 2013

AbstractBacteriostatic and bactericidal activities of rifampicin, isoniazid, streptomycin, enviomycin and ethambutol against Mycobacterium tuberculosis, Mycobacterium avium—M. intracellulare complex and Mycobacterium kansasii were studied in different growth phases. Bacteriostatic activities of the drugs were similar in different growth phases, except isoniazid. M, tuberculosis was much less susceptible to isoniazid in the lag phase than in the log and the stationary phases. In contrast, bactericidal activity was influenced by the growth phase. M. tuberculosis was killed by isoniazid, streptomycin and rifampicin. The bactericidal activity of isoniazid was strongest. The bactericidal activity of isoniazid and streptomycin was most marked in the log phase. M. avium complex and M. kansasii resisted the bactericidal activity, but some strains of M. avium complex were killed by streptomycin and enviomycin, and the activities of these two drugs were most marked in the lag phase.
Proposal of a New Scheme for the Serological Typing of Enterococcus faecalis Strains
Authors: Maekawa Shizue , Yoshioka Migaku , Kumamoto Yoshiaki 
Published: November 2013

AbstractNo systematic study on serotyping of Enterococcus faecalis has been reported since 1964 when M.E. Sharpe conducted serotyping of group D streptococcus in U.K. So, we attempted to re‐evaluate serotyping of E. faecalis. For this purpose, we received 42 Sharpe's strains and first examined for their biochemical characteristics as E. faecalis. Only 9 of the 42 strains were identified as E. faecalis. We raised rabbit antisera against a large number of E. faecalis strains, including the 9 Sharpe's strains, 2 strains obtained from CDC in U.S.A. and 36 strains isolated from patients hospitalized in different cities of Japan. From the results of cross‐agglutination tests and absorption tests performed on these antisera using a large number of E. faecalis strains, we were able to classify 21 distinct serotype strains and to prepare 21 mono‐specific typing antisera by absorption of the antisera to the type strains with appropriate cross‐agglutinating strains. When 832 E. faecalis strains were serotyped with the 21 typing antisera, 638 strains (76.7%) were typable. Thus, we propose a provisional scheme of 21 distinct serovars in E. faecalis.
Phage Pattern and Antibiotic Resistance Pattern of Coagulase‐Negative Staphylococci Obtained from Immunocompromised Patients
Authors: Barcs István , Herendi Ágnes , Lipcsey András , Bognár Csaba , Hashimoto Hajime 
Published: November 2013

AbstractA total of 152 coagulase‐negative staphylococcal strains were isolated from clinical samples of 14 patients hospitalized after bone‐marrow transplantation in a specialized hospital ward in Hungary, during an 18‐month period between 1987 and 1989. Two species, Staphylococcus epidermidis and Staphylococcus haemolyticus, predominated (each, 45%). Using Pulverer and co‐workers' phage set for typing, 68% of the isolates were typable; 16 phage patterns were observed. A characteristic long pattern with phages Ph10/Ph13/Ph15/U4/U15/U16/U20/U33/U46 appeared only in S. epidermidis, among 5 of 11 colonized patients (8.5% of all strains). Single lysis with phage Phl3 was observed in 7 of the 14 patients (49% of all strains), in species S. capitis, S. epidermidis, S. haemolyticus, S. hominis, and S. warneri. In S. haemolyticus, non‐typable strains predominated (66%); this character occurred only in 2% among other species. The strains colonizing the immunocompromised patients differed from each other in phage pattern, antibiotic resistance pattern, and/or slime production. No hospital infection was suggested. On the other hand, high incidence of two well‐definable phage patterns raises some relationship between phage receptors or some regulatory systems in phage multiplication and factors responsible for special colonization as common surface properties.
Pathogenicity of Bacteroides fragilis Group in Rat Intra‐Abdominal Abscesses
Authors: Jotwani Ravi , Gupta Usha , Watanabe Kunitomo , Ueno Kazue 
Published: November 2013

AbstractAttempts were made to study the pathogenicity of some strains of Bacteroides fragilis group in the rat intra‐abdominal abscess model. Multiple intra‐abdominal abscesses were produced in 50 to 70% of animals when an inoculum containing 109 CFU/ml of any of the five species of Bacteroides fragilis group was injected. Rising homologous antibody titers determined by indirect fluorescent antibody test were observed till the 3rd week when tested last, indirectly confirming the multiplication of the organisms as also evident by viable count of bacteria in the abscesses. In some cases in addition to inoculated organisms some intestinal bacteria like Escherichia coli, Proteus mirabilis and Streptococcus spp. were also recovered from the abscess pus. Studies with the electron microscope showed presence of capsular polysaccharide only in Bacteroides fragilis and Bacteroides thetaiotaomicron. It was doubtful in Bacteroides distasonis and absent in Bacteroides ovatus and Bacteroides vulgatus, suggesting that virulence factor beside the capsular polysaccharide may be playing a role. Further studies are required to investigate the virulence factor responsible for the pathogenicity of noncapsulated species.
Mechanisms of Augmented Resistance of Cyclosporin A‐Treated Mice to Influenza Virus Infection by Trehalose‐6, 6′‐Dimycolate
Authors: Sazaki Katsuhiko , Yoshida Itsuro , Azuma Masanobu 
Published: November 2013

AbstractCyclosporin A (CsA), which is an immunosuppressive drug of helper T lymphocytes, diminished a resistance of mice to influenza virus infection. Mice inoculated intravenously with trehalose‐6,6′‐dimycolate (TDM, a glycolipid component of the cell wall of Mycobacterium) in an oil‐in‐water emulsion (TDM emulsion) recovered the resistance to influenza virus infection impaired by CsA. Number of antibody‐producing cells was markedly reduced in CsA‐ and/or TDM‐treated mice. Interferon production in lung of TDM‐treated mice was augmented; however, it was extremely reduced not only in CsA‐treated mice, but also in CsA‐ and TDM‐treated mice. Activities of natural killer cells of CsA‐ and/or TDM‐treated mice were not different from that of control mice. Numbers of lymphocytes in lung of TDM‐treated mice and CsA‐ and TDM‐treated mice were more predominantly increased than that of control mice. Analysis of lung lymphocytes by flow cytometry revealed no difference between the populations of L3T4+ T lymphocytes and Lyt‐2.2+ T lymphocytes in CsA‐ and/or TDM‐treated mice and the populations in control mice. However, the population of γδ T cell receptor positive (γδ TCR+) lymphocytes increased markedly in lung of TDM‐treated mice and also CsA‐ and TDM‐treated mice. In vitro experiments showed that macrophage cultures treated with TDM emulsion released a mediator(s) which activates T lymphocytes, but not B lymphocytes. These and our earlier results suggest that the recovered anti‐influenza virus resistance of CsA‐treated mice by treatment with TDM emulsion was caused by elicitation of macrophages with TDM, then activation of T lymphocytes, especially γδ TCR+ lymphocytes.
Serological Cross‐Reaction between Intact and Chemically Modified Lipopolysaccharides of O1 Vibrio cholerae Inaba and Non‐O1 V. cholerae Bio‐Serogroup Hakata
Authors: Isshiki Yasunori , Haishima Yuji , Kondo Seiichi , Hisatsune Kazuhito 
Published: November 2013

AbstractSerological cross‐reaction of intact as well as chemically modified LPS from O1 Vibrio cholerae 569B (Inaba) with non‐O1 V. cholerae Hakata LPS, which contain α(1→2)‐linked N‐acetyl perosamine‐homopolymer constituting their O polysaccharide chain, was studied by passive hemolysis test by using their LPS as antigen for sensitizing sheep red blood cells (SRBC). The N‐deacylation of the α(1→2)‐linked linear 3‐deoxy‐tetronyl perosamine‐homopolymer constituting the O polysaccharide chain in 569B LPS resulted in virtual elimination of their serological reactivity with both homologous Inaba and heterologous Hakata antisera. Furthermore, when the resultant NH2 groups of the N‐deacylated perosamine‐homopolymers in 569B LPS were N‐acylated with acetyl, propionyl or butanoyl groups, they markedly recovered the serological reactivity to a marked extent, in particular, their pronounced cross‐serological reactivity with heterologous Hakata antiserum. These results are believed to be compatible with the interpretation that the Inaba antigen factor C possessed by the two bacteria studied is related to the common occurrence of the N‐acyl groups, regardless of what the acyl groups are, residing in the perosamine residues of the perosamine‐homopolymers constituting the O polysaccharide chain of their LPS.
cDNA Cloning of an Aspartic Proteinase Secreted by Candida albicans
Authors: Mukai Hiroyuki , Takeda Osamu , Asada Kiyozo , Kato Ikunoshin , Murayama Somay Yamagata , Yamaguchi Hideyo 
Published: November 2013

AbstractcDNA of an aspartic proteinase secreted by Candida albicans No. 114 was isolated using the polymerase chain reaction (PCR). The primary structure of the enzyme was deduced from the nucleotide sequence of the cDNA and compared with the structures of Saccharomyces cerevisiae proteinase A and vacuolar aspartyl proteinase of C. albicans. The mature aspartic proteinase consisted of 341 amino acid residues, and was 17.6 and 15.3% identical with the proteinase A and the aspartyl proteinase, respectively. Two active aspartic acid sites and the amino acids near those sites were conserved in the aspartic proteinase. We also showed that there is another gene of aspartic proteinase than that of strain ATCC10231 reported by Hube et al (J. Med. Vet. Mycol. 29 (1991)) in the same C. albicans genome, both in that strain and in No. 114.
An Improved Method for the Serotyping of Free Coagulase from Staphylococcus aureus
Authors: Tajima Yutaka , Nagasawa Zenzo , Tanabe Ichiro , Yamada Hisashi , Kusaba Koji , Tadano Jutaro 
Published: November 2013

AbstractThe serotyping of free coagulase, one of the most reliable ways to identify strains of Staphylococcus aureus, and widely employed in Japan, has been improved by adding magnetite sand to the reaction mixture. Culture medium supernatant and a type‐specific antibody are mixed in a well of a microtiter plate, and plasma‐enriched bovine fibrinogen is treated with magnetite sand. The use of tranexamic acid and gum arabic in the reaction mixture also increases the sensitivity of the reaction. Finally, the plate is placed on a magnetic stirrer. If the type of the coagulase corresponds to that of the antibody, no clot formation will occur, and this is easily confirmed by the movement of the sand. Although the amount of reaction mixture required is much less than that for the conventional tube method, our new method is able to detect slight increases in viscosity of the reaction mixture due to fibrin formation even before complete clotting occurs, thus providing very high sensitivity. Clot formation can also be judged by observing a turbid mass of fibrin in the well (Hwang's method), but this approach is a little slower than our method involving immobilization of magnetite sand.
Cloning of the Genes That Control Formation of the Fimbrial Colonization Factor Antigen III (CFA/III) from an Enterotoxigenic Escherichia coli
Authors: Shinagawa Hideo , Taniguchi Tooru , Yamaguchi Osamu , Yamamoto Koichiro , Honda Takeshi 
Published: November 2013

AbstractEnterotoxigenic Escherichia coli strain 260‐1 produces colonization factor antigen III and heat‐labile enterotoxin. A 55‐kb plasmid controlling the expression of the colonization factor antigen was isolated from this strain after it was labeled with ampicillin resistance transposon, Tn3. When this plasmid was introduced into E. coli K‐12 strains, it induced the formation of pili that were morphologically and immunologically identical to those on the surface of 260‐1 cells, as examined by electron microscopic observation and with the specific antiserum. The physical map of the plasmid was constructed, and the 17.4‐kb region was found to be responsible for the expression of the pili.
A Factor Found in Aged Tungstate Solution Enhanced the Antibacterial Effect of β‐Lactams on Methicillin‐Resistant Staphylococcus aureus
Authors: Tajima Yutaka , Nagasawa Zenzo , Tadano Jutaro 
Published: November 2013

AbstractWe have found a factor (Factor T) in aged mixtures of tungstate and phosphate which greatly enhances the antibacterial effects of β‐lactams on both inducible and constitutive methicillin‐resistant Staphylococcus aureus, but not on methicillin‐susceptible S. aureus. Factor T alone did not strongly inhibit bacterial growth. There was no synergism of Factor T with other classes of antibiotics, nor with other groups of bacteria, and it reduced the efficacy of aminoglycosides and tetracycline. Upon preparation of Factor T, acidifying and heating the mixture of tungstate and phosphate resulted in a high yield and reproducibility, and no enhancing effect was observed when other anions such as sulfate or molybdate were used instead. Factor T is heat‐ and acid‐stable but labile to alkalization, and is probably a complex of phosphate and tungstate.
Suppressive Effect of Liposomes Containing DNA Coding for Diphtheria Toxin A‐Chain on Cells Transformed with Bovine Leukemia Virus
Authors: Kakidani Hitoshi , Watarai Shinobu , Onuma Misao , Tomochika Ken‐ichi , Yasuda Tatsuji 
Published: November 2013

AbstractA recombinant plasmid which contained a gene for diphtheria toxin A‐chain (DT‐A) under the control of the long terminal repeat (LTR) of bovine leukemia virus (BLV) (BLV‐LTR) was constructed to test a novel application of liposomes as antiviral agents. The promoter activity of BLV‐LTR was estimated by the chloramphenicol acetyltransferase (CAT) assay using a plasmid which contains the coding sequence of CAT under the control of BLV‐LTR (pBLVCAT). When BLV‐infected cells were transfected with pBLVCAT, CAT activity was detected. BLV‐uninfected cell lines, however, showed no detectable CAT activity. The plasmid DNA entrapped in liposomes was added to BLV‐infected cells in culture. Syncytium formation induced by BLV‐infected cells was effectively suppressed by the liposomes containing the gene for DT‐A under the control of BLV‐LTR. Conversely, liposomes containing the gene for DT‐A without a promoter showed no such effect. DT‐A gene‐containing liposomes with BLV‐LTR did not affect formation of syncytium induced by bovine immunodeficiency virus. These observations indicate that BLV‐infected cells were readily targeted on the level of gene expression. This strategy could be applied to the treatment of BLV‐induced B‐cell proliferation of cattle, and further to other viral/neoplastic diseases where specific gene expression is exerted.
Characterization of a Phenol Oxidase from Cryptococcus neoformans var. neoformans
Authors: Ikeda Reiko , Shinoda Takako , Morita Takashi , Jacobson Eric S. 
Published: November 2013

AbstractIn Cryptococcus neoformans, enzymic oxidation of various catechols leads to melanin, a proposed virulence factor. A phenol oxidase enzyme of Cryptococcus neoformans var. neoformans produced at 25 C has been purified from an ultracentrifugal supernatant of an extract of broken cells. Hydrophobic interaction chromatography followed by anion‐exchange column chromatography allowed purification of the phenol oxidase. The molecular weight of the enzyme estimated by gel filtration was about 80,000 and a dimeric species (Mw = 160,000) was suggested. The isoelectric point of the protein was approximately 4.1. An NH2‐terminal 31 amino acid sequence was determined using phenol oxidase electroblotted onto a PVDF membrane after nondenaturing gel electrophoresis. Upon searching the Peptide Institute (Osaka) data base, no proteins with high degrees of homology were found.
Genomic Analysis of Borrelia japonica Sp. Nov. Isolated from Ixodes ovatus in Japan
Authors: Kawabata Hiroki , Masuzawa Toshiyuki , Yanagihara Yasutake 
Published: November 2013

AbstractGenetic characteristics of 12 Borrelia isolates from the tick, Ixodes ovatus, I. persulcatus, and the rodent, Apodemus speciosus ainu, in Japan were compared to members of the three genospecies of Borrelia burgdorferi sensu lato; B. burgdorferi sensu stricto, B. garinii and group VS461. The methods used in this study were the quantitative microplate DNA hybridization assay and restriction fragment length polymorphism (RFLP) analyses of the flagellin structural genes and the 16S rRNA genes. The six isolates from I. persulcatus and A. speciosus ainu were identified as genospecies B. garinii using RFLP analysis of the flagellin and 16S rRNA genes. In contrast, RFLP analysis of the six isolates from I. ovatus indicated that they were different from the three reported genospecies. DNA homology studies confirmed the RFLP results. The six isolates from I. ovatus had DNA homologies ranging from 85 to 99%, whereas DNA relatedness of the I. ovatus isolate with strains belonging to the three genospecies was 50 to 64%. These results suggest that the strains isolated from I. ovatus in Japan differ from the three genospecies and should be classified as a new genospecies of B. burgdorferi sensu lato. We propose that strains isolated from I. ovatus should be classified as B. japonica sp. nov.
Cytotoxic Effect of Hemolytic Culture Supernatant from Enterococcus faecalis on Mouse Polymorphonuclear Neutrophils and Macrophages
Authors: Miyazaki Shuichi , Ohno Akira , Kobayashi Intetsu , Uji Tatsuya , Yamaguchi Keizo , Goto Sachiko 
Published: November 2013

AbstractWe reconfirmed that the LD50S of hemolytic Enterococcus faecalis strains were significantly less than those of nonhemolytic E. faecalis strains in normal mice. Hemolysin produced by E. faecalis lysed human, horse, rabbit, and mouse erythrocytes, but not cow and sheep erythrocytes. Sphingomyelin comprises a part of the lipid composition of the erythrocyte membrane of all mammalian species tested. But phosphatidylcholine exists only in human, horse, rabbit, and mouse. These two lipids inhibited lysis of horse erythrocytes by hemolytic E. faecalis. Phosphatidylcholine is probably the binding component on the membrane of erythrocytes for E. faecalis hemolysin. The hemolytic culture supernatant lysed not only erythrocytes but also mouse polymorphonuclear neutrophils (PMNs) and macrophages.