bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genotyping of Pneumocystis carinii f. sp. hominis Isolates in Japan Based on Nucleotide Sequence Variations in Internal Transcribed Spacer Regions of rRNA Genes
Authors: Hosoya Noriaki , Takahashi Takashi , Wada Miki , Endo Tokiomi , Nakamura Tetsuya , Sakashita Hiroyuki , Kimura Kyoko , Ohnishi Kenji , Nakamura Yoshikazu , Mizuochi Tsuguo , Iwamoto Aikichi 
Published: November 2013

AbstractGenotyping of Pneumocystis carinii (Pc) isolated from 24 bronchoalveolar lavage (BAL) fluid specimens in Japan was examined based on nucleotide sequence variations in internal transcribed spacer regions 1 and 2 (ITS1 and ITS2, respectively) of rRNA genes. We found 11 ITS1 genotypes including 2 novel ones and 11 ITS2 genotypes including 3 new ones. Combining the ITS1 and ITS2 genotypes resulted in 30 ITS genotypes, of which 10 are newly described in this report. Two or more genotypes in ITS regions in a specimen were observed in 16 of 24 patients. Our results will be of help for the epidemiological investigation of Pc infection.
Detection of Microsporum canis in the Skin Scrapings and Hairs of Dogs with Dermatophytosis Based on Sequences of the Chitin Synthase 1 Gene
Authors: Kano Rui , Nakamura Yuka , Watanabe Shinichi , Hasegawa Atsuhiko 
Published: November 2013

AbstractIn the present study, to confirm Microsporum canis infection rapidly, we detected the chitin synthase gene 1 (CHS1) gene of M. canis in the hair and skin samples of four dogs with dermatophytosis. Amplification of the DNAs in the four samples with CHS1 primers yielded fragments of about 620‐bp. Nucleotide sequence analysis of the CHS1 gene fragments from samples and a reference strain of M. canis gene showed more than 99% similarity. The method presented in this study can rapidly detect the DNA of M. canis in skin scrapings, and we anticipate that it will be a useful microbiological tool for the diagnosis of M. canis infections in animals and humans.
Firm Adherence of Staphylococcus aureus and Staphylococcus epidermidis to Human Hair and Effect of Detergent Treatment
Authors: Mase Kiyomi , Hasegawa Tadao , Horii Toshinobu , Hatakeyama Kazuto , Kawano Yasushi , Yamashino Takafumi , Ohta Michio 
Published: November 2013

AbstractStaphylococcus aureus and S. epidermidis are common pathogens in hospitals, and care should be taken not to disseminate these organisms among patients. We have focused on human hair as a source of bacterial contamination. We treated hair with culture solutions of S. aureus and S. epidermidis, and then performed scanning electron microscopy. Bacteria were detected on the surface of the cuticles of the hair, and the attached bacteria were not completely removed even by repeated washing with detergents. These results suggested that hair could be a source of bacterial contamination and indicated the importance of decontamination of hair.
Identification of Amino Acids of Influenza Virus HA Responsible for Resistance to a Fusion Inhibitor, Stachyflin
Authors: Yoshimoto Jun , Kakui Mayumi , Iwasaki Hiroko , Sugimoto Hirohiko , Fujiwara Tamio , Hattori Naohiko 
Published: November 2013

AbstractWe have recently described a novel hemagglutinin (HA) conformational change inhibitor of human influenza virus, Stachyflin (Yoshimoto et al, Arch. Virol., 144, 1–14, 1999). Stachyflin‐resistant variants of human influenza A/WSN/33 (H1N1) virus were isolated in vitro and the nucleotide sequences of their HA genes were determined. The relation of amino acid substitutions and Stachyflin resistance was analyzed with in vitro membrane fusion between HA‐expressing cells and octadecylrhodamine (R18)‐labelled chick erythrocytes (RBC). The amino acid substitutions, lysine to arginine at position 51 or lysine to glutamic acid at position 121 of the HA2 subunit of the HA protein was enough to confer a Stachyflin‐resistant phenotype of HA protein. The molecular mechanism of anti‐HA conformational change activity of Stachyflin is discussed.
Expression of Recombinant Capsid Proteins of Chitta Virus, a Genogroup II Norwalk Virus, and Development of an ELISA to Detect the Viral Antigen
Authors: Kobayashi Shinichi , Sakae Kenji , Suzuki Yasumoto , Ishiko Hiroaki , Kamata Kunio , Suzuki Kenji , Natori Katsuro , Miyamura Tatsuo , Takeda Naokazu 
Published: November 2013

AbstractThe second open reading frame (ORF2) gene of the Chitta virus (CHV) was cloned to construct a recombinant baculovirus. The CHV ORF2 is predicted to encode a capsid protein of 535 amino acids (aa). CHV showed a high aa identity in the capsid region with genogroup II Norwalk virus (NV) (65–85%), but a low aa identity with genogroup I NV (44–46%). Phylogenetic analysis of the ORF2 gene demonstrated that CHV is genetically closely related to the Hawaii virus included in genogroup II NV. The recombinant capsid protein of CHV (rCHV) self‐assembled to form empty virus‐like particles (VLPs) when expressed in insect cells with the recombinant baculovirus. An enzyme‐linked immunosorbent assay (ELISA) based on antisera to rCHV was developed to detect CHV antigen in stools. The antigen ELISA appeared to be highly specific to both rCHV and CHV‐like strains. In addition, combined use of antigen ELISAs using antibodies against two antigenically distinct recombinant VLPs, the recombinant Chiba virus (rCV) and recombinant Seto virus (rSEV), enabled us to determine the genetic as well as antigenic relationship among these three viruses.
Determination of the Protective Effects of Neutralizing Anti‐Hepatitis B Virus (HBV) Immunoglobulins by Epitope Mapping with Recombinant HBV Surface‐Antigen Proteins
Authors: Park Jung‐hyun , Cho Eun‐wie , Lee Yun‐jung , Shin Song Yub , Kim Kil Lyong 
Published: November 2013

AbstractAnti‐hepatitis B virus (HBV) surface‐antigen immunoglobulins prepared from human sera are clinical reagents which have been approved for prophylactic treatment in HBV‐exposed persons. The passive immunoprophylaxis with immunoglobulins is meant to cross‐link viral particles, which are then further cleared by the host's own immune system. While antibodies specific for both anti‐S‐ and anti‐preS proteins have been proved to serve as effective anti‐viral agents, so far the fine antigen specificity of clinical immunoglobulin preparations has not been determined. Using recombinant proteins covering the hepatitis B surface antigen, in the present study, the specificity of a commercially available immunoglobulin preparation was determined and immunodominant epitopes were mapped. Here, it is shown that the major reactivity of anti‐HBV immunoglobulins is directed against the S‐protein, and that no reactivity to the preS2 but a weak binding activity to the preS1 region was detectable. The antigen reactivity within the preS1 region was biased to the C‐terminal region, which indicates the presence of a putative B‐cell epitope. The evaluation of the antigen specificity and determination of novel protective epitopes will provide valuable information for the further development and improvement of prophylactic HBV immunoglobulins.
First Isolation of Stephanoascus ciferrii from a Cat
Authors: Kano Rui , Makimura Koichi , Kushida Toshiaki , Nomura Megumi , Yamaguchi Hideo , Hasegawa Atsuhiko 
Published: November 2013

AbstractThe present study deals with the first isolation of Stephanoascus ciferrii from a cat. A 2‐year‐old female Persian cat weighing 2.25 kg was referred to an animal hospital with a chief complaint of otitis externa of the left ear. Microscopic examination of specimen from the left ear disclosed yeast cells. The colony of the clinical isolate was cream‐colored, rough, raised and wrinkled. The microscopic examination of the clinical isolate revealed abundant branched and septated mycelia with small ramified chains of oval blastoconidia, variable in size, and arranged alongside the hyphae. Amplification of the isolate DNA with LSU rDNA primers yielded a fragment of about 570 bp, whose nucleotide sequence of the isolate showed 100% similarity to that of Stephanoascus ciferrii in the GenBank database. Therefore, the isolate was identified as Stephanoascus ciferrii, confirming the result of mycological examination by molecular analysis.
Molecular Cloning and Partial Characterization of Rat Procarboxypeptidase R and Carboxypeptidase N
Authors: Kato Tomomi , Akatsu Hiroyasu , Sato Tomoo , Matsuo Seiichi , Yamamoto Takayuki , Campbell William , Hotta Nigishi , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractCarboxypeptidase R (EC 3.4.17.20) (CPR) and carboxypeptidase N (EC 3.4.17.3) (CPN) cleave carboxy‐terminal arginine or lysine residues from biologically active peptides such as kinins or anaphylatoxins in the circulation thereby regulating their activities. Although CPN is present in a stable active form in plasma, CPR is generated from proCPR, a plasma zymogen, by proteolytic enzymes such as thrombin, thrombin‐thrombomodulin complex and plasmin. We have isolated rat proCPR and CPN cDNA clones which can induce enzymatic activities in culture supernatants of the transfected cells. mRNA of proCPR was detected only in rat liver by Northern hybridization and showed hepatocyte‐specific expression. Expression of proCPR mRNA was enhanced following LPS injection, indicating that proCPR production is increased under inflammatory conditions.
Upregulated Expression of Iba1 Molecules in the Central Nervous System of Mice in Response to Neurovirulent Influenza A Virus Infection
Authors: Mori Isamu , Imai Yoshinori , Kohsaka Shinichi , Kimura Yoshinobu 
Published: November 2013

AbstractThe present study deals with the expression of Ibal molecules, a novel EF‐hand Ca2+‐binding protein, in the brain after stereotaxic introduction of the neurovirulent WSN strain of influenza A virus into the olfactory bulb of C57BL/6 mice. The virus selectively targeted the paraventricular and anterior olfactory nuclei. Infected neurons appeared as early as at day 3 post infection and degenerated and vanished by day 12. The Ibal molecule was normally expressed in resting microglia. The overexpression of the Ibal in microglial cells was detected at day 3 post infection, culminating at day 7 with a morphological activation. Ibal‐immunopositive macrophages outnumbered microglia in the paraventricular and anterior olfactory nuclei, where the infected neurons had degenerated. Macrophages totally disappeared by day 12, and the Ibal‐expression in microglia was reduced to a normal level by day 35. Lack of perforin predisposed the mice to long‐term virus infection of the brain, leading to the prolonged Ibal‐overexpression. These results show that the Ibal is upregulated in the mouse brain in response to influenza virus infection and may play significant roles in the regulation of some immunological and pathophysiological functions of microglia during virus infection.
Lack of Evidence of an Association between the Carriage of Virulence Plasmid and the Bacteremia of Salmonella typhimurium in Humans
Authors: Chiu Cheng‐hsun , Chu Chishih , Ou Jonathan T. 
Published: November 2013

AbstractThe involvement of the virulence plasmid (pSTV) of Salmonella typhimurium in human salmonellosis was examined. Most of the 224 clinical strains isolated from the blood (53) and nonblood samples (171) contained a 90 kb or larger plasmid, most of which were pSTV. The rates of pSTV carriage in the isolates showed no statistically significant difference between those derived from the blood and those from other sources (87% vs. 83%; χ2=0.49, 0.1 < P < 0.9), suggesting that pSTV may not play a critical role in promoting S. typhimurium bacteremia in humans. Nine strains with representative plasmid profiles were tested for the mouse virulence. The result revealed that these clinical isolates contained all three virulent types known: the avirulent, the highly virulent when a pSTV was present, and the moderately virulent regardless of the presence or absence of pSTV. This indicated that mouse virulence of S. typhimurium did not correlate their virulence in humans. Clinical data showed that most patients with primary bacteremia had underlying immunosuppressive diseases, whereas only a few patients with secondary bacteremia had preexisting diseases (87% vs. 13%; χ2 = 22.73, P < 0.005). It is suggested that the contribution of pSTV to S. typhimurium bacteremia in humans is likely to be limited, and both the host factor and the microbial virulence determinants on the chromosome are more important than virulence plasmid in predisposing patients to bacteremia.
Localization of Major, High Molecular Weight Proteins in Bacteroides forsythus
Authors: Higuchi Naoya , Murakami Yukitaka , Moriguchi Keiichi , Ohno Norikazu , Nakamura Hiroshi , Yoshimura Fuminobu 
Published: November 2013

AbstractBacteroides forsythus produces species‐specific major proteins with high molecular weights of 270 and 230‐kDa (270K and 230K). A specific antibody raised against 270K was used for Western blot analysis and immunoelectron microscopy. Western blot analysis showed that the 270K and 230K proteins were immunologically similar. Immunogold labeling of ultrathin‐sectioned bacterial cells and biochemical fractionation revealed that these proteins were localized at the outermost cell surface, not in the cytoplasm. These results suggest that major proteins ubiquitous to this species may form the S‐layer.
Th1‐Th2 Cytokine Kinetics in the Bronchoalveolar Lavage Fluid of Mice Infected with Cryptococcus neoformans of Different Virulences
Authors: Abe Koh , Kadota Jun‐ichi , Ishimatsu Yuji , Iwashita Tetsuji , Tomono Kazunori , Kawakami Kazuyoshi , Kohno Shigeru 
Published: November 2013

AbstractTh1 immune response plays an important role in protection against infection with Cryptococcus neoformans in mice. We investigated the effect of virulence of C. neoformans on cytokine production in the lung of a mouse model of pulmonary cryptococcosis. BALB/c mice were inoculated intratracheally with a high or low virulence strain of C. neoformans, followed by serial measurements of Th1 and Th2 cytokine concentrations in the bronchoalveolar lavage (BAL) fluid using appropriate enzyme‐linked immunosorbent assay kits. The number of colony‐forming units (CFU) increased with time, and all mice infected with the highly virulent strain were dead at 28 days after inoculation. In contrast, the number of microorganisms diminished with time in the mice infected with the low virulence strain during the 4‐week study. The numbers of neutrophils and lymphocytes in the BAL fluid paralleled those of CFU. High neutrophil counts were observed in the BAL fluid of mice infected with the highly virulent strain, while lymphocyte counts were increased only in the later part of the study in mice infected with the high and low virulence strains. The concentrations of Th2 cytokine, interleukin (IL)‐4 were significantly higher in mice infected with the highly virulent strain at days 14 and 21 of infection, whereas the level of Th1 cytokine, interferon‐gamma, was significantly higher in the latter strain at days 7 and 14. Our results suggest that strain‐specific difference in the organism's ability to induce (or evade) the host immune system contributes to the outcome of infection.
Emergence of New Influenza A Viruses Which Carry an Escape Mutation of the HLA‐B27‐Restricted CTL Epitope of NP in Japan
Authors: Kashiwagi Takahito , Hamada Nobuyuki , Iwahashi Jun , Hara Koyu , Ueda Tatsuo , Noguchi Hidetaro , Toyoda Tetsuya 
Published: November 2013

AbstractInfluenza A viruses (H3N2) isolated in 1998 in Nagasaki, Japan, carried a mutation (384R→G) in one of the anchor amino acids of the HLA‐B27‐restricted cytotoxic T lymphocyte (CTL) epitope of NP (383–391). Phylogenetic analysis revealed that these viruses have been isolated only in Japan to date and belong to the unique lineages.
Control of Immunologically Crossreactive Leptospiral Infection by Administration of Lipopolysaccharides from a Nonpathogenic Strain of Leptospira biflexa
Authors: Matsuo Kouki , Isogai Emiko , Araki Yoshio 
Published: November 2013

AbstractIn our previous paper (Matsuo, K., Isogai, E., and Araki, Y., Carbohydr. Res., 328: 517–524, 2000), antigenic polysaccharides obtained from the lipopolysaccharide (LPS) fraction of a nonpathogenic leptospira, Leptospira biflexa patoc Patoc I, are shown to be broadly crossreactable with most rabbit antisera elicited by immunization with various pathogenic leptospires. The result led us to test a protective effect of the same LPS in a hamster model system by heterologously challenging with a pathogenic leptospira, L. interrogans manilae UP‐MMG. Firstly, a similarity in the antigenic epitopes of L. biflexa and L. interrogans was confirmed by the following assays. In the microscopic agglutination test (MAT), a hamster antiserum elicited by immunization with the L. biflexa‐LPS preparation was shown to agglutinate cells of L. interrogans. Contrarily, in the enzyme‐linked immunosorbent assay (ELISA), the L. biflexa‐LPS preparation was shown to crossreact with a hamster antiserum elicited by immunization with whole cells of L. interrogans. These results suggest that the same or closely related antigens may be present on the cell surfaces of both L. biflexa patoc Patoc I and L. interrogans manilae UP‐MMG. Furthermore, in a protective assay, the prior administration of a L. biflexa‐LPS preparation resulted in raising a protective response in hamsters against challenge by L. interrogans without any side effect. The protective effect was strongly dependent on the dose amounts and/or administration times of L. biflexa‐LPS. Thus, L. biflexa‐LPS preparations can use as a potent vaccine against leptospirosis caused by various leptospires.
Effect of Bicozamycin on the Eradication of Shiga Toxin‐Producing Escherichia coli in Calves
Authors: Misawa Naoaki , Sueyoshi Masuo , Uemura Ryoko , Kakemizu Yoshihiro , Kawashima Kumiko , Nagatomo Hiroshi , Kondo Fusao , Murakami Takayuki , Takahashi Yoshihiro 
Published: November 2013

AbstractFifty‐nine calves, aged 11 days to 9 months, from three farms breeding Japanese Black beef cattle in Miyazaki Prefecture, Japan, were examined for the presence of Shiga toxin‐producing Escherichia coli (STEC). A high prevalence of STEC was detected among calves, with 45 (76.3%) animals carrying STEC including different serogroups (O26, O74, O111, O114, O119, O127, O153, O157, and ONT) and toxin types. The number of STEC in the feces was estimated by a combined method involving enumeration of colony‐forming units by a plate‐most‐probable‐number (plate‐MPN) technique and polymerase chain reaction for the detection of Shiga toxin genes. Fecal shedding ranged from 101 to 10s MPN/g feces. To evaluate the safety and efficacy of bicozamycin (BCM: previously named as bicyclomycin) in eradicating STEC, 30 calves carrying STEC with or without diarrhea were examined. Fifteen calves were treated orally with BCM (10 mg/kg/day) once daily for 5 days, and the other 15 were untreated. Twenty‐four hours after the last dose, fecal specimens were collected from both groups to compare the number of coliforms and STEC with those before treatment. BCM‐treated animals had a significantly lower number of coliforms and STEC compared to the untreated calves. The STEC eradication rate was 86.6% (13/15) in the BCM‐treated group, compared to 0% (0/15) in the control group. The corresponding cure rates for diarrhea were 87.5 (7/8) and 0% (0/3), respectively. No adverse reactions were observed in the calves during treatment. It is concluded that BCM is an effective agent for the eradication of STEC in calves with or without diarrhea.
Apoptosis Observed in Murine Peritoneal Macrophages Treated with Interferon γ through Staphylococcal Enterotoxin‐Dependent Cell‐Mediated Cytotoxicity
Authors: Sakurada Junji , Tajima Akiko , Shinji Hitomi , Seki Keiko , Masuda Shogo 
Published: November 2013

AbstractThe concept of superantigens is well‐known and widely accepted. In this brief communication, we analyze the behaviour of antigen‐presenting cells after T‐cell activation by staphylococcal enterotoxin B, a representative superantigen. We tried to activate murine T cells by inflammatory mouse peritoneal macrophage in the presence of staphylococcal enterotoxin B, but no T‐cell activation was observed. We, therefore, analyzed surface‐specific antigens of the macrophages. They expressed insufficient amounts of MHC class II, CD80 and CD86 molecules on the surface of the cells. On the contrary, increased amounts of MHC class II and CD86 molecules on the cell surfaces were observed after incubation with interferon γ. Interferon γ‐primed macrophages were found to be competent to activate T cells in the presence of staphylococcal enterotoxin B. To our surprise, these macrophages underwent apoptosis in parallel with T‐cell activation.
Size Conversion of a Linear Plasmid in the Relapsing Fever Agent Borrelia duttonii
Authors: Takahashi Yukie , Cutler Sally J. , Fukunaga Masahito 
Published: November 2013

AbstractBorreliae have genomes composed of both linear and circular replicons. We have characterized the organization of linear DNA molecules from the Borrelia duttonii strain Ly. It contains a linear one megabase chromosome and 12 linear plasmids of 11 to 200 kb in size. A variant of the strain obtained after successive in vitro cultivation in BSKII medium had a 69 kb molecule instead of the 44 kb linear plasmid. No detectable differences in the growth rates and cellular structures were found. Southern hybridization using the vsp33 gene sequence from Borrelia hermsii as a probe showed that both plasmids (69 and 44 kb molecules) contained a similar part of the sequence. The spirochetes of the parental strain cause erythrocytes to aggregate in mice blood, but the variant did not form such aggregates and seemed to have lost its infectivity in mice. Size conversion of the linear plasmid may be associated with the host‐parasite relationship in mammals.
The Significance of Cytomegalovirus Infection over the Clinical Course of Adult T‐Cell Leukemia/Lymphoma
Authors: Fujiwara Hiroshi , Eizuru Yoshito , Matsumoto Tadashi , Kukita Toshimasa , Imaizumi Rie , Kawada Hideaki , Ohtsubo Hideo , Matsushita Kakushi , Arima Naomi , Tei Chuwa 
Published: November 2013

AbstractThe significance of cytomegalovirus (CMV) infections developed over the clinical course of adult T‐cell leukemia/lymphoma (ATLL) were evaluated in relation to the patient survival rate, ATL activity and immunocompetent cells. ATLL patients with CMV infections on admission exhibited a poor survival rate, while patients with CMV infections at any time after admission survived longer than those not infected with this virus. ATLL patients who exhibited a numbers of CMV infection on admission showed higher ATL activity and had lower numbers of CD8‐positive and CD56‐positive cells than those who developed CMV infections at any time after admission. Therefore, it appears likely that patients with CMV infections on admission were in an immunosuppressive state due to aggressive ATL activity.