bluuebunny/crossref_metadata_2025_split huggingface dataset:
Multiplication of Legionella pneumophila in HeLa Cells in the Presence of Cytoskeleton and Metabolic Inhibitors
Authors: Goldoni Paola , Cattani Lorena , Carrara Stefania , Pastoris Maddalena Castellani , Sinibaldi Laura , Orsi Nicola 
Published: November 2013

AbstractA study has been carried out on the action of cytoskeleton and metabolic inhibitors on intracellular multiplication in HeLa cells of a virulent strain of Legionella pneumophila serogroup 6. The effects of the substances were separately tested on both penetration and intracellular multiplication of L. pneumophila. Only cytochalasin A and 2‐deoxy‐d‐glucose (2dG) affected bacterial internalisation, whereas intracellular multiplication was inhibited by cytochalasins A, B, C, D and J (D being the most active) and by 2dG with a dose‐response effect. The action of 2dG was counteracted by 50 mM glucose. Experiments carried out with cytochalasin D and a rhodamine‐phalloidin conjugate showed the involvement of cytoskeletal elements in intracellular multiplication of Legionella; compounds acting on microtubules had no effect.
Relation between Reactivity to the NS‐4 Region Peptides of Hepatitis C Virus (HCV) and Clinical Features among Patients Infected with HCV Genotype 1b
Authors: Sakugawa Hiroshi , Nakasone Hiroki , Nakayoshi Tomofumi , Kawakami Yuko , Kinjo Fukunori , Saito Atsushi , Nakayoshi Tomokuni , Yamashiro Akihiro 
Published: November 2013

AbstractNearly all patients infected with hepatitis C virus (HCV) genotype 1b have reactivity to the core (c22‐3) or non‐structural (NS)‐3 region (c33c) protein in a second‐generation recombinant immunoblot assay (RIBA‐2). However, reactivities to the NS‐4 region antigens (5‐1‐1, c100‐3) vary among patients. To clarify whether differences in serological reactivities to the NS‐4 antigens are associated with the clinical features or response to interferon (IFN) therapy of patients infected with hepatitis C virus (HCV) genotype 1b, we clinically investigated 115 such patients. Positive reactions to 5‐1‐1 and c100‐3 were seen in 75.7 and 79.1%, respectively, of the patients. There were no differences between the patients with and those without antibodies to NS‐4 region antigens (5‐1‐1, c100‐3) with regard to age, duration of HCV infection, severity of liver disease and virus load. Fifty‐one of the patients were treated with recombinant IFN‐α, and 17 of the 51 patients showed sustained response to the therapy. The sustained response was more frequently seen in the patients positive for antibodies to both 5‐1‐1 and c100‐3 as compared with those negative for either or both antibodies (41.0% vs. 8.3%, P < 0.05).
Detection of HIV‐Gag p24‐Specific Antibodies in Sera and Saliva of HIV‐1‐Infected Adults and in Sera of Infants Born to HIV‐1‐Infected Mothers
Authors: Yasuda Sachiko , Iwasaki Masahiro , Oka Shin‐ichi , Naganawa Satoshi , Nakasone Tadashi , Honda Mitsuo , Sata Tetsutaro , Kojima Asato , Matsuda Syunji , Takemori Toshitada , Tsunetsugu‐Yokota Yasuko 
Published: November 2013

AbstractSecretory immunoglobulin A (IgA) is known to play an important role in the mucosal defense against a variety of pathogens. Although the role of IgA antibodies during sexual transmission of HIV is not clear, HIV‐specific IgA antibodies have been detected in various mucosal secretions of HIV‐infected individuals. Using a monoclonal antibody against human IgA, we established an ELISA system to detect anti‐HIV p24 IgA antibodies in sera and saliva. We have analyzed the levels of anti‐HIV p24 IgG and IgA antibodies in sera and saliva of 107 and 119 adults, respectively, with HIV infection at different clinical stages, and in the sera of 13 infants born to HIV‐infected mothers. The level of anti‐HIV p24 IgA antibodies was lower in sera and higher in saliva as compared to that of anti‐HIV p24 IgG antibodies. Where the percentage of HIV‐specific serum antibody‐positive cases decreased with disease progression, that of saliva antibody‐positive cases increased in AIDS patients. Among the 13 infants born to HIV‐infected mothers, 7 infants were HIV‐p24‐specific serum IgA positive. These sera were negative for anti‐HIV p24 secretory IgA, suggesting that some infants develop their own immune responses against HIV infection. Thus, the detection of HIV‐specific IgA antibodies, especially in saliva, could be a simple and reliable test for the diagnosis of HIV infection.
Inhibitory Effect of Bacterial Attachment on Candidal Growth Due to Adherence with Mannose‐Sensitive Pili
Authors: Seki Keiko , Kobayashi Kiyoshi , Tomiya Yoshinori , Sakurada Junji , Murai Miyo , Usui Akemi , Masuda Shogo 
Published: November 2013

AbstractA bacterial strain with affinity to Candida albicans was successfully obtained from a natural environment. An uncovered Petri dish containing a suspension of heat‐killed C. albicans cells was allowed to stand in a laboratory for several days. Some bacteria which had adhered to the candidal cells were tested for their ability to agglutinate the cells. A bacterial strain, designated later as CAB‐1, was found to agglutinate candidal cells through bridging by mannose‐sensitive pili. CAB‐1 showed similar bacteriological characteristics to those of Citrobacter freundii by ID test. The adherence of CAB‐1 to candidal cell was precisely presented by scanning electron microscopy. The inhibitory effect of CAB‐1 attachment to candidal cells on the growth of Candida was also preliminarily confirmed.
Systemic Dissemination by Intrarectal Infection with Listeria monocytogenes in Mice
Authors: Nishikawa Shinsuke , Hirasue Mayuko , Miura Tomisato , Yamada Kyogo , Sasaki Sanae , Nakane Akio 
Published: November 2013

AbstractOrally ingested Listeria monocytogenes is known to penetrate into Peyer's patches (PP) and translocate to the spleen and liver. Herein, extraintestinal dissemination of the bacterium independent of PP was investigated. Dissemination of Listeriae to the spleen and liver was observed in intrarectally infected mice as well as in intragastrically infected animals in spite that no Listeriae were detected in the small intestines of mice infected intrarectally. Decreased numbers of intestinal intraepithelial lymphocytes (iIEL) and increased numbers of lymphocytes in the contents of the small and large intestines were observed after intragastric infection and in the large intestine after intrarectal infection, giving the assumption that the leakage of iIEL caused by injury of epithelial layers in intestines might occur during infection. These results suggest that L. monocytogenes might be able to disseminate through small and large intestines in part by a PP‐independent mechanism.
Pathogenicity of a Mutant Friend Spleen Focus‐Forming Virus Encoding an Env‐Like Membrane Glycoprotein (gp55) with Substitution by a Xenotropic Murine Leukemia Virus Env gp70 Sequence
Authors: Yugawa Takashi , Amanuma Hiroshi 
Published: November 2013

AbstractFriend spleen focus‐forming virus (F‐SFFV) is a replication‐defective acutely leukemogenic mouse retrovirus and encodes an envelope protein (Env)‐like membrane glycoprotein (gp55) in its defective env gene, which is responsible for the early stage of the viral leukemogenesis. Gp55 is a modified Env protein and contains a polytropic mink cell focus‐inducing (MCF) murine leukemia virus (MuLV) Env gp70‐derived sequence in its amino‐terminal region. To evaluate the possibility that the presumed binding of gp55 to an MCF MuLV receptor protein has some role in leukemogenesis, we examined the biological activities of a mutant gp55 (XE gp55), which has a xenotropic MuLV Env gp70 amino‐terminal region. XE gp55 displayed almost the same biological activities as the wild‐type gp55, excluding the above possibility.
Detection of Verotoxin‐Producing Escherichia coli O157:H7 by Multiplex Polymerase Chain Reaction
Authors: Nagano Isao , Kunishima Miyuki , Itoh Yoshihiro , Wu Zhiliang , Takahashi Yuzo 
Published: November 2013

AbstractWe constructed primers for multiplex polymerase chain reaction (PCR) to detect verotoxin‐producing Escherichia coli (VTEC) O157:H7. The multiplex PCR primers were designed from the sequence of the flagellin structural gene of Escherichia coli flagellar type H7 (GenBank under accession number L07388), and from the sequence of the rfbE gene of Escherichia coli O157:H7 (GenBank under accession number S83460). In addition to these primers, we used a primer pair reported by Karch and Meyer (J. Clin, Microbiol. 27: 2751‐2757, 1989) to amplify various VT genes from VTEC. All of the examined specimens (18 isolates) of VT‐producing E. coli O157:H7 showed a positive result by the multiplex PCR test with the three sets of primers. The sensitivity of detection for VT‐producing E. coli O157:H7 was shown to be at least 3,000 cells per PCR tube.
Emendation of GenusAchromobacterandAchromobacter xylosoxidans(Yabuuchi and Yano) and Proposal ofAchromobacter ruhlandii(Packer and Vishniac) Comb. Nov.,Achromobacter piechaudii(Kiredjian et al.) Comb. Nov., andAchromobacter xylosoxidansSubsp.denitrificans(Rüger and Tan) Comb. Nov.
Authors: Yabuuchi Eiko , Kawamura Yoshiaki , Kosako Yoshimasa , Ezaki Takayuki 
Published: November 2013

AbstractBased on the results of GC content determination and 16S rRNA sequence analysis among the type strains ofAchromobacter xylosoxidans, 4Alcaligenesspecies, 5Bordetellaspecies, and 12 species of 4 other genera, the separation of genusAchromobacterYabuuchi and Yano 1981, with the type speciesAchromobacter xylosoxidans, is confirmed.Alcaligenes ruhlandii(Packer and Vishniac) Aragno and Schlegel 1992 is a distinct species and not a senior synonym ofAchromobacter xylosoxidans. Alcaligenes ruhlandiiandAlcaligenes piechaudiiKiredjian et al 1986 are transferred to genusAchromobacter. Thus 2 new combinations,Achromobacter ruhlandii(Packer and Vishniac) andAchromobacter piechaudii(Kiredjian et al) are proposed; their type strains are ATCC 15749 and ATCC 43552, respectively.Alcaligenes denitrificansRüger and Tan 1983 is also transferred to genusAchromobacterand ranked down to the subspecies ofAchromobacter xylosoxidans. Thus a new subspecies name,Achromobacter xylosoxidanssubsp.denitrificans(Rüger and Tan) is proposed. The type strain of the subspecies is ATCC 15173. This proposal automatically creates type subspecies,Achromobacter xylosoxidanssubsp.xylosoxidans, with type strain ATCC 27061. An emended description of genusAchromobacterand of type speciesAchromobacter xylosoxidansare given.
First Report of Trichosporon ovoides Isolated from the Home of a Summer‐Type Hypersensitivity Pneumonitis Patient
Authors: Sugita Takashi , Nishikawa Akemi , Ikeda Reiko , Shinoda Takako , Sakashita Hiroyuki , Sakai Youko , Yoshizawa Yasuyuki 
Published: November 2013

AbstractTrichosporon species have been known to cause summer‐type hypersensitivity pneumonitis (SHP). During the isolation of yeasts from an SHP patient's house, we recovered a strain belonging to the genus Trichosporon. Morphologically, the isolate produced rectangular arthroconidia when grown on corn meal agar. DNA‐DNA hybridization experiments identified the isolate as T. ovoides. A slide agglutination test using specific factor sera demonstrated that the serotype of the strain was type II. Previously, T. asahii, a serotype II species, was considered to be the major antigen of SHP, but it is possible that T. ovoides may also be responsible for SHP. This is the first report of T. ovoides isolated from an SHP patient's home environment.
Trimethoprim Resistance and Susceptibility Genes in Staphylococcus epidermidis
Authors: Totake Kyoko , Kobayashi Nobuharu , Odaka Takeshi 
Published: November 2013

AbstractGenes encoding trimethoprim (TMP)‐resistant and ‐susceptible dihydrofolate reductases (DHFR) in Staphylococcus epidermidis isolated in Saitama Prefecture were compared with the TMP‐resistant DHFR gene of S. aureus, dfrA. The nucleotide sequences of TMPr and TMPS genes in five S. epidermidis isolates tested could be divided into three types: type 1, identical with the TMPr gene dfrA that had been found in S. aureus; type 3, identical with the TMPs gene dfrC in S. epidermidis; and type 2, having only two nucleotide substitutions to dfrC with no amino acid change. TMPr isolates carried either one of the type 2 or type 3 sequences in addition to the type 1 sequence. A Southern hybridization analysis revealed that, in TMPr S. epidermidis, the type 1 sequence was located on a 5.5 kb EcoRI‐EcoRV restriction fragment together with the sequence for the gentamicin (GM)‐resistant gene, while the type 2 or type 3 sequence was located on the 1.0 kb EcoRI‐EcoRV fragment. No plasmid‐carrying dfrA‐homologous sequence was detected in the S. epidermidis isolates we tested. These results suggest that the TMPr and GMr genes are closely linked and located on the chromosome in S. epidermidis isolated in Japan.
Chemical and Immunological Characterization of a Low Molecular Weight Outer Membrane Protein of Salmonella typhi
Authors: De Andrade Carlos Maurício , Ferreira Antônio Gomes Pinto , Da Silva Joana D'Arc Cardoso , Nascimento Hilton Jorge , Junior José Godinho Da Silva 
Published: November 2013

AbstractA new immunogenic outer membrane protein, Omp‐28 (MW 28,000 and pI 4.6), was isolated from smooth Salmonella typhi cells by the use of an extracting medium containing 6 m urea, 1% deoxycholate and 5 mM EDTA. The purification of Omp‐28 was performed by gel filtration and fast ion exchange chromatography. This protein showed to be the prevalent component isolated by the latter methodology. Omp‐28 is formed by three identical subunits (MW 9,000), not linked by disulfide bonds. The partial N‐terminal amino acid sequence of Omp‐28 presented great homology with part of the sequence of an Escherichia coli protein found in a precursor whose sequence was predicted by c‐DNA. ELISA and Western blotting identified Omp‐28 as the major antigenic protein present in the outer membrane protein fraction, isolated by gel filtration. Antibodies against Omp‐28 were detected by ELISA in 43% of 28 sera from typhoid fever convalescent patients. The antisera from mice immunized with Omp‐28 and the highest positive typhoid fever convalescent serum gave a positive bactericidal test, killing 50% of Salmonella typhi cells in serum dilutions of 1/80 and 1/320, respectively. These results indicate the immunogenic importance of Omp‐28 isolated from Salmonella typhi outer membrane and strongly suggest it should be used in further studies of animal protection against the disease caused by this pathogenic bacteria.
Mechanism of Membrane Damage by Clostridium perfringens Alpha‐Toxin
Authors: Nagahama Masahiro , Michiue Kei , Mukai Masakazu , Ochi Sadayuki , Sakùrai Jun 
Published: November 2013

AbstractThe effect of Clostridium perfringens alpha‐toxin on liposomes prepared from phosphatidylcholine (PC) containing the fatty acyl residues of 18 carbon atoms was investigated. The toxin‐induced carboxyfluorescein (CF) leakage and phosphorylcholine release from multilamellar liposomes increased as the phase transition temperature of the phosphatidylcholines containing unsaturated fatty acyl residues decreased. However, there was no difference between the sensitivity of the different phosphatidylcholines solubilized by deoxycholate to the phospholipase C (PLC) activity of the toxin. However, the toxin did not hydrolyze solubilized distearoyl‐l‐α‐phosphatidylcholine (DSPC) or phosphatidylcholine containing saturated fatty acyl residue, and caused no effect on liposomes composed of DSPC. These results suggest that the activity of the toxin is closely related to the membrane fluidity and double bond in PC. The N‐terminal domain of alpha‐toxin (AT1‐246) and variant H148G did not induce CF leakage from liposomes composed of dioleoyl‐l‐α‐phosphatidylcholine (DOPC). H148G bound to the liposomes, but AT1‐246 did not. However, the C‐terminal domain (AT251‐370) conferred binding to liposomes and the membrane‐damaging activity on AT1‐246. These observations suggest that the membrane‐damaging action of alpha‐toxin is due to the binding of the C‐terminal domain of the toxin to the double bond in the PC in the bilayer and hydrolysis of the PC by the N‐terminal domain.
Single‐Step Reverse Transcription‐Polymerase Chain Reaction for the Detection of Hepatitis C Virus RNA
Authors: Mizutani Tetsuya , Ikeda Masanori , Saito Satoru , Sugiyama Kazuo , Shimotohno Kunitada , Kato Nobuyuki 
Published: November 2013

AbstractWe developed a novel single‐step reverse transcription‐polymerase chain reaction (RT‐PCR), which is equal in sensitivity and specificity to RT‐nested PCR, based on both reverse transcriptase and Taq DNA polymerase working efficiently under single buffer reaction conditions. Using in vitro synthesized hepatitis C virus (HCV) RNA, it was demonstrated that 10‐100 copies of HCV RNA could be detected with a set of primers that amplify a 144 base‐pair sequence unique to the 5′‐noncoding region of HCV RNA. Furthermore, this method was successfully performed on serum and liver biopsy specimens obtained from patients with chronic hepatitis C. In addition, HCV RNA from in vitro HCV‐infected MT‐2C cells, which supported HCV replication, was also detected by this method. The method is anticipated to improve the detection of small amounts of RNA, such as that of HCV, promoting both labor savings and the prevention of carry‐over contamination.
Lipopolysaccharide (LPS) Stimulates the Production of Tumor Necrosis Factor (TNF)‐α and Expression of Inducible Nitric Oxide Synthase (iNOS) by Osteoclasts (OCL) in Murine Bone Marrow Cell Culture
Authors: Kikkawa Ichiro , Saito Shinji , Tominaga Kaoru , Hoshino Yuichi , Ooi Yoshio , Nakano Masayasu 
Published: November 2013

AbstractOsteoclasts (OCL) resorb bone. They are essential for the development of normal bones and the repair of impaired bones. The function of OCL is presumed to be supported by cytokines and other biological mediators, including tumor necrosis factor (TNF)‐α and nitric oxide (NO). Bacterial lipopolysaccharide (LPS) is a potent inducer of TNF‐α and inducible nitric oxide synthase (iNOS), which is the specific enzyme for synthesizing NO from L‐arginine. To obtain direct evidence on LPS‐induced TNF‐α production and iNOS expression by OCL, OCL‐enriched cultures were prepared by 7‐day cocultures of bone marrow cells of adult BALB/c mice and osteoblastic cells (OBs) derived from calvaria of newborn BALB/c mice, and the generation of TNF‐α and iNOS in OCL stimulated with LPS was examined immunocytochemically. When the cultured cells were stimulated with 100 ng/ml of LPS, OCL clearly showed TNF‐α and iNOS expression. Without LPS‐stimulation, no expression was observed. TNF activity in the culture supernatants of the OCL‐enriched cultures in the presence of LPS was also detected by cytotoxic assay that used TNF‐sensitive L929 cells. The dentin resorption activity of OCL was estimated by area and number of pits formed on dentin slices, which were covered by the OCL fraction and cultured in the presence or absence of LPS, sodium nitroprusside (SNP; a NO generating compound), NG‐monomethyl L‐arginine acetate (L‐NMMA; a competitive inhibitor of NO synthase (NOS)), or LPS plus L‐NMMA. Pit formation was obviously inhibited in the presence of SNP and slightly inhibited in the presence of L‐NMMA, but it was not affected in the presence of LPS or LPS plus L‐NMMA. These findings indicate that OCL produces TNF and expresses iNOS in response to LPS, but the LPS‐activation of OCL scarcely affects pit formation by them.
Phylogenetic Analysis of the Genus Bifidobacterium and Related Genera Based on 16S rDNA Sequences
Authors: Miyake Taeko , Watanabe Koichi , Watanabe Tsugio , Oyaizu Hiroshi 
Published: November 2013

AbstractThe 16S rRNA gene sequences were determined for type strains of 21 Bifidobacterium species. A phylogenetic tree was constructed using the determined sequences and sequences from DNA databases, which contain the sequences of 11 type strains of Bifidobacterium species and 11 strains of related genera. All species of the genus Bifidobacterium and Gardnerella vaginalis ATCC 14018 belonged to a cluster phylogenetically distinct from the other genera. The cluster was divided into two subclusters: subcluster 1 composed of most species of Bifidobacterium and G. vaginalis, and subcluster 2 consisting of two species, B. denticolens and B. inopinatum; both of which were isolated from human dental caries. In the genus Bifidobacterium, four groups of species are known to be moderately to highly related by DNA‐DNA hybridization. The four groups of species exhibited more than 99% similarity among their 16S rDNA sequences within each group. These results indicated that species with around 99% or more similarity in their 16S rDNA sequences should be confirmed for species identities.
Serological Studies of an Acid‐Labile O‐Polysaccharide of Proteus vulgaris OX19 Lipopolysaccharide Using Human and Rabbit Antibodies
Authors: Kaca Wieslaw , Swierzko Anna ST. , Ziolkowski Andrzej , Amano Ken‐ichi , Senchenkova Sof'Ya N. , Knirel Yuriy A. 
Published: November 2013

AbstractIn a Weil‐Felix test, sera from patients infected with Rickettsia sp. agglutinate Proteus OX types of bacteria and Proteus lipopolysaccharide (LPS) are responsible for the cross‐reaction. Data on the character of LPS of one of the OX group strains, Proteus vulgaris OX19, are contradictory, and it remained unclear whether it has an O‐polysaccharide (OPS) and is thus LPS of the smooth type (S) or not (rough‐type LPS). Our studies showed that P. vulgaris OX19 (strain PZH‐24) produces a smooth‐type LPS that contains a long‐chain OPS, but it undergoes depolymerization during mild acid hydrolysis conventionally used for LPS delipidation and loses the serological activity. An elucidation of the complete structure of OPS demonstrated the presence of a glycosyl phosphate linkage responsible for the acid‐lability of the polysaccharide chain. In ELISA, both IgM type antibodies in a Weil‐Felix test with human anti‐Rickettsia typhi sera and rabbit anti‐P. vulgaris OX19 antibodies reacted with OPS. Rabbit antibodies did not inhibit the cross‐reaction with human antibodies and thus bind to different epitopes.
Analysis of Genetic Relationships and Antimicrobial Susceptibility of Verotoxin‐Producing Escherichia coli Strains Isolated in Nagasaki Prefecture, Japan in 1996
Authors: Izumikawa Koichi , Hirakata Yoichi , Yamaguchi Toshiyuki , Yoshida Ryoji , Nakano Michiko , Matsuda Junichi , Mochida Chikako , Maesaki Shigefumi , Tomono Kazunori , Yamada Yasuaki , Tashiro Takayoshi , Kohno Shigeru , Kamihira Shimeru 
Published: November 2013

AbstractA total of 19 Escherichia coli O157 isolates were obtained in Nagasaki Prefecture, in the southwestern part of Japan, between 1990 and 1996. Pulsed‐field gel electrophoresis (PFGE) and computer‐assisted analysis were applied to determine genetic relationships among these strains. Fragment patterns of the isolates in Nagasaki, as determined by PFGE, were compared with those of isolates in other areas where large outbreaks and sporadic cases of E. coli O157 infection occurred. Similarity values of all the strains isolated in Nagasaki Prefecture were over 0.65 except for E. coli O26. Some strains were identical to the strains isolated from the areas where large outbreaks occurred. All strains were susceptible to ampicillin, fosfomycin, minocycline, amikacin, ofloxacin and sulfamethoxazole‐trimethoprim.