bluuebunny/crossref_metadata_2025_split huggingface dataset:
Theiler's Murine Encephalomyelitis Virus (TMEV) Subgroup Strain‐Specific Infection in Neural and Non‐Neural Cell Lines
Authors: Obuchi Masatsugu , Yamamoto Jiro , Uddin Md. Nasir , Odagiri Takato , Iizuka Hideaki , Ohara Yoshiro 
Published: November 2013

AbstractGDVII subgroup strains of Theiler's murine encephalomyelitis virus (TMEV) are highly virulent and produce acute polioencephalomyelitis in mice. Neither viral persistence nor demyelination is demonstrated in the few surviving mice. In contrast, DA subgroup strains are less virulent and establish a persistent central nervous system infection which results in demyelinating disease. We previously reported a subgroup‐specific infection in a macrophage‐like cell line, J774‐1 cells; i.e., GDVII strain does not replicate in J774‐1 cells, whereas the DA strain actively replicates in these cells. In addition, this subgroup‐specific virus growth is shown to be related to the presence of L* protein, a 17 kDa protein translated out‐of‐frame of the viral polyprotein from an AUG located 13 nucleotides downstream from the polyprotein's AUG. The present paper demonstrated that this subgroup‐specific infection is observed in murine monocyte/macrophage lineage cell lines, but not in other murine cell lines including neural cells. An RNase protection assay also suggested that L* protein‐related virus growth is regulated at the step of viral RNA replication. As macrophages are reported to be the major cell harboring virus during the chronic demyelinating stage, the activity of L* protein with respect to virus growth in macrophages may be a key factor in clarifying the mechanism(s) of TMEV persistence, which is probably a trigger to spinal cord demyelination.
Prevalence of the Virulence Plasmids of Nontyphoid Salmonella in the Serovars Isolated from Humans and Their Association with Bacteremia
Authors: Chiu Cheng‐Hsun , Lin Tzou‐Yien , Ou Jonathan T. 
Published: November 2013

AbstractTo determine if the virulence plasmid is one of the elements contributing to Salmonella bacteremia in humans, 436 clinical Salmonella isolates of different serovars were examined by a specific multiplex polymerase chain reaction assay for the presence of a virulence plasmid. These serovars showed differences in their ability to produce particular disease syndrome in humans. In the serovars usually causing bacteremia without concomitant gastroenteritis (primary bacteremia), i.e., S. choleraesuis, S. dublin, and S. enteritidis in this study, the rate of virulence plasmid carriage was 100%, while among those occasionally generating bacteremia following an episode of gastroenteritis (secondary bacteremia), the majority were plasmidless. Only a portion of S. typhimurium strains harbored a virulence plasmid; however, the rates of virulence plasmid carriage in S. typhimurium were not statistically different between non‐fecal and fecal isolates (90% vs. 85%, 0.1 < P < 0.9). These results indicate that the virulence plasmids may be important for primary bacteremia, but not secondary bacteremia, to occur.
Alteration of Cell Wall Composition Leads to Amphotericin B Resistance in Aspergillus flavus
Authors: Seo Kuniko , Akiyoshi Hiroto , Ohnishi Yoshinari 
Published: November 2013

AbstractAn amphotericin B (AmB)‐resistant mutant was isolated from a wild‐type AmB‐susceptible strain of Aspergillus flavus by serial transfer of conidia on agar plates containing stepwise increased concentrations of AmB up to 100 μtg ml−1. The acquired resistance of my celia was specific for polyene‐antibiotics AmB, nystatin and trichomycin. Spheroplasts derived from the resistant mycelia were as susceptible to AmB as the wild‐type. Chemical analysis of the cell wall revealed that levels of alkali‐soluble and ‐insoluble glucans were significantly higher in the resistant mycelia as compared to those in the wild‐type. When resistant mycelia were treated with SDS, they adsorbed as much AmB as wild‐type mycelia. These results suggest that alterations in the cell wall components of mycelia, especially 1,3‐α‐glucan and protein complex in the outermost wall layer, lead to AmB resistance in A. flavus.
Characterization of Mouse DAF on Transfectant Cells Using Monoclonal Antibodies Which Recognize Different Epitopes
Authors: Ohta Rieko , Imai Masaki , Fukuoka Yoshihiro , Miwa Takashi , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractSeveral membrane proteins prevent host cells from homologous complement attack. In humans, one such protein, decay‐accelerating factor (DAF), exists as two isoforms, a GPI anchored form and a secreted form, which are generated by alternative splicing. DAF in mouse is also expressed as two isoforms, a GPI anchored form (GPI‐DAF) and a transmembrane form (TM‐DAF), which are produced from two separate genes. In this study, we transfected cDNA of mouse GPI‐DAF or TM‐DAF into Chinese hamster ovary (CHO) cells. Both isoforms of DAF on CHO cells were shown to regulate mouse complement C3 deposition mediated by the classical and alternative pathways and the inhibitory activity of both isoforms was species restricted. The two mouse DAF isoforms were effective against rat complement but not against human and guinea pig complement. Furthermore, we produced hamster mAbs to mouse DAF using GPI‐DAF transfectant cells and established seven unique mAbs (RIKO‐1–7). Western blotting analysis using RIKO‐3, which reacts with both GPI‐DAF and TM‐DAF, and RIKO‐4, which is an anti‐GPI‐DAF specific mAb, indicated that GPI‐DAF was expressed on erythrocytes, spleen and testis, and that TM‐DAF was expressed only in testis.
Increased Sensitivity of Cholera Toxin Detection by Enzyme‐Linked Immunosorbent Assay with Chemical Immobilization of Antibody onto Nylon Surface Providing Hydrophobicity
Authors: Koike Norio , Yabushita Yasunori , Yan Zhang Dong , Lida Tetsuya , Honda Takeshi 
Published: November 2013

AbstractWe developed an improved method to chemically immobilize antibodies on a nylon surface using a styrene maleic anhydride copolymer having an aryl group, which provides hydrophobicity to the nylon surface. We applied it to a modified enzyme‐linked immunosorbent assay (slip‐ELISA) for the detection of cholera toxin (CT). The sensitivity of slip‐ELISA for CT detection was 1,000 times higher than that of conventional methods of physical adsorption using polystyrene plates, and 10 times higher than that of the method of chemical immobilization using maleic anhydride methylvinyl ether copolymer.
Comparison of T‐Cell Receptor Jβ Gene Usage in Spleen Cells of Different Mouse Strains
Authors: Kato Tomohiro , Sasakawa Hiroko , Kurokawa Manae , Masuko‐Hongo Kayo , Hamada Chikuma , Yamamoto Kazuhiko , Nishioka Kusuki 
Published: November 2013

AbstractTo elucidate whether T‐cell receptor Jβ gene usage was affected by major histocompatibility complex haplotypes and other genetic backgrounds, we investigated such usage with Jβ‐specific probes in four different mouse strains. As a result, (a) frequent usage of Jβ2.1 and Jβ2.6, (b) infrequent usage of Jβ1.3, Jβ1.5 and Jβ1.6, and (c) predominant usage of the Jβ2 cluster compared to the Jβ1 cluster were found. Importantly, these biases were common to almost all the tested Vβ families of the four strains. Thus, TCR Jβ usage would be independent of the major histocompatibility complex haplotypes and other genetic backgrounds.
Serum Antibody Level against GroEL Type Heat‐Shock Protein of Campylobacter jejuni in Patients with Guillain‐Barré Syndrome
Authors: Fujimoto Shuji , Fujita Masaki 
Published: November 2013

AbstractRecently there has been an increase in the number of cases reported of Guillain‐Barré syndrome (GBS) developed after Campylobacter jejuni infection. To investigate the role of a C. jejuni GroEL‐type heat‐shock protein (CjHsp60) in the infection and induction of GBS, we examined the antibody level against CjHsp60 in 27 human sera, including GBS and non‐GBS patients, by an enzyme‐linked immunosorbent assay. Sera from patients with C. jejuni infection, despite the development of GBS, had a higher titer of anti‐CjHsp60 antibody than those of patients without the infection and healthy control subjects. The patients with C. jejuni infection followed by GBS had slightly higher levels of this antibody than did the patients with infection who did not develop GBS, but there was no statistical significance. In conclusion, CjHsp60 is found to be one of the major immunogenic antigens in actual C. jejuni infection, but no evidence that supports the direct relationship between this protein and C. jejuni‐associated GBS was found in this study.
Assessment of Mucosal Immune Response in Genitourinary Tract Using Urine
Authors: Kohno Takeyuki , Tanaka Hiroko , Watabe Kazuhito , Yamashita Shinji , Sezaki Hitoshi , Nadai Tanekazu , Sugie Yohko , Ogouchi Toshiyo 
Published: November 2013

AbstractA novel method to assess mucosal immune response in the genitourinary mucosa after immunization with a mucosal vaccine has been developed. In this method, secretory IgA antibody is measured by a highly sensitive enzyme immunoassay (immune‐complex transfer enzyme immunoassay) using urine as a specimen. The urinary IgA antibody response could be detected by the immune‐complex transfer enzyme immunoassay. In contrast, a conventional enzyme immunoassay (enzyme‐linked immunosorbent assay (ELISA)) could not detect this response because of its low sensitivity. Because urine samples can be collected easily and nontraumatically, not only from experimental animals but also from humans, both males and females, the present method may be applicable for assessing the protective efficacy of candidates for mucosal vaccines against sexually transmitted microorganisms, such as human immunodeficiency virus. Furthermore, the usefulness of this method for novel mucosal vaccine formulae was shown for a model in which vaccine antigen and Bordetella pertussis adjuvant were adsorbed onto CaCO3 and enclosed in enteric coated capsules.
Induction of Interleukin 8 (IL‐8) Production by Pseudomonas Nitrite Reductase in Human Alveolar Macrophages and Epithelial Cells
Authors: Sar Borann , Oishi Kazunori , Matsushima Kouji , Nagatake Tsuyoshi 
Published: November 2013

AbstractInterleukin‐8 (IL‐8) participates in the generation of dense neutrophil accumulations in bronchopulmonary infections caused by Pseudomonas aeruginosa (P. aeruginosa). We have recently reported that nitrite reductase, a bifunctional enzyme located in the periplasmic space of P. aeruginosa, induces IL‐8 generation in bronchial epithelial cells (K. Oishi et al. Infect. Immun. 65: 2648 2655, 1997). We examined whether or not Pseudomonas nitrite reductase (PNR) could also stimulate human alveolar macrophages (AM) and pulmonary type II epithelial‐like cells (A549) to induce IL‐8 production and mRNA expression as well as the production of TNFα and IL‐1β. We demonstrated a time‐ and dose‐dependent IL‐8 protein synthesis and IL‐8 mRNA expression, but no TNFα or IL‐1β production, by A549 cells in response to PNR. New protein translation was not required for PNR‐mediated IL‐8 mRNA expression in the same cells. Furthermore, simultaneous stimulation of PNR with serial doses of TNFα or IL‐1β resulted in additive IL‐8 production in A549 cells. In adherent AM, PNR enhanced IL‐8 protein synthesis and IL‐8 mRNA expression in a time‐dependent fashion. PNR similarly induced a time‐dependent production of TNFα and IL‐1β by human adherent AM. Neutralization of TNFα or IL‐1β did not influence the levels of IL‐8 production in adherent AM culture. We also evaluated whether the culture supernatants of the A549 cells or AM stimulated with PNR could similarly mediate neutrophil migration in vitro. When anti‐human IL‐8 immunoglobulin G was used for neutralizing neutrophil chemotactic factor (NCF) activities in the culture supernatants of these cells stimulated with 5 μg/ml of PNR, the mean percent reduction of NCF activities were 49‐59% in A549 cells and 2434% in AM. Our present data support that PNR directly stimulates AM and pulmonary epithelial cells to produce IL‐8. PNR also mediates neutrophil migration, in part, through IL‐8 production from AM and pulmonary epithelial cells. These data suggest the contribution of PNR to the pathogenesis of bronchopulmonary infections due to P. aeruginosa.
Quantification of Phagocytosis in Human Neutrophils by Flow Cytometry
Authors: Heinzelmann Michael , Gardner Sarah Appel , Mercer‐Jones Mark , Roll Amy J. , Polk Hiram C. 
Published: November 2013

AbstractPhagocytosis represents a central element of the host response to microbial invasion. We describe a flow cytometric method for measuring the kinetics of phagocytosis of two bacteria by human polymorphonuclear leukocytes (PMNs). Over a 60‐min period, isolated human PMNs were exposed to Staphylococcus aureus (rapidly phagocytosed) and Klebsiella pneumoniae (slowly phagocytosed). This method distinguished adherent from ingested bacteria by quenching fluorescein isothiocyanate‐labeled extracellular bacteria with ethidium bromide. This further allowed the exclusion of dead, highly permeable, and subsequently bright‐red fluorescent PMNs. Our experiments with two different bacteria, various PMN‐to‐bacteria ratios (1:1, 1:10, 1:100), and different individuals proved that 1) flow cytometric analysis is accurate and useful for characterizing phagocytosis, 2) adherent bacteria can be distinguished from ingested bacteria after quenching with ethidium bromide, and that 3) phagocytosis kinetics of two bacteria with different onsets of phagocytosis can be determined by flow cytometry and the assessment of a score that quantifies phagocytosis.
Modulation of Barrier Function of Small Intestinal Epithelial Cells by Lamina Propria Fibroblasts in Response to Lipopolysaccharide: Possible Role of TNFα in Inducing Barrier Dysfunction
Authors: Chakravortty Dipshikha , Kumar K. S. Nanda 
Published: November 2013

AbstractRecent evidence suggests an interaction between immune, enteric neural and fibroblasts in the regulation of intestinal function. Earlier, we have reported that lipopolysaccharide (LPS) induced cell proliferation, collagen synthesis and production of proinflammatory mediators in lamina propria fibroblasts. In this report, we investigated the change in transepithelial resistance (TER) as a marker of epithelial barrier function by lipopolysaccharide (LPS) and its modulation by human small intestinal lamina propria fibroblasts (HSILPF). Epithelial cells incubated with LPS alone did not show any change in the TER at any concentration or prolonged exposure. However, co‐cultivation of epithelial cells with lamina propria fibroblasts which had been exposed to LPS resulted in a rapid decrease in TER by 2 hr. The decrease in the TER was continued till 8 hr followed by returning to the basal level by 24 hr. The supernatant of LPS‐treated HSILPF was less effective in causing a fall in the TER than HSILPF itself. The fall in TER was accompanied by loosening of tight junctions as depicted by increased penetration of horse radish peroxidase (HRP) across the epithelial cells from the apical to the basal side. Increased incorporation of 3[H]thymidine (tritiated thymidine) in epithelial cells was observed at 48 hr in the presence of LPS‐treated HSILPF. The decrease in TER during the early time period in epithelial cells was abrogated to 70% by incubating the LPS‐treated HSILPF and the conditioned medium of LPS‐treated HSILPF with anti‐TNFα antibody, and not with antibody to other cytokines like IL1α, IL1β, IL6 and IL8. Overall, these results suggest that TNFα produced by HSILPF in response to LPS as a soluble form cause a decrease in the TER and loosening of tight junctions, and such early changes in the epithelial barrier may contribute to local inflammation in the gut.
Ribotyping of Vibrio parahaemolyticus Isolates Obtained from Food Poisoning Outbreaks in Taiwan
Authors: Wong Hin‐chung , Ho Chia‐yun , Kuo Li‐ping , Wang Tien‐kuei , Lee Chi‐lung , Shih Daniel Yang‐chih 
Published: November 2013

AbstractVibrio parahaemolyticus is a prevalent food‐borne pathogen in Taiwan, Japan and other Asian countries. This work presents a novel ribotyping method for the molecular epidemiological examination of this pathogen. Genomic DNA was fragmented by HindIII digestion and hybridized with cDNA probe for Escherichia coli 16S and 23S RNA genes. A total of 121 isolates obtained from outbreaks during 1992 and 1994 in Taiwan were characterized by this ribotyping method. Four to seventeen restricted fragments were visualized in these isolates. After hierarchical cluster analysis, these isolates were grouped into thirty different ribotypes. In addition, A3, A7, E3 and F1 were the major ribotypes, consisting of 22.3, 13.2, 9.1, and 8.3% of the isolates, respectively. A, E, F, G and B were the major groups, consisting of 46.2, 14.0, 9.1, 6.7, and 6.7% of the isolates, respectively. The discriminatory ability of this ribotyping method, as determined by Simpson's index of diversity, was 0.93, which closely resembled that of a previously reported pulsed‐field gel electrophoresis method.
The Expression of the Pathogenic Yeast Candida albicans Catalase Gene in Response to Hydrogen Peroxide
Authors: Nakagawa Yoshiyuki , Watanabe Koji Koide, Kiyomi , Morita Yasuhiro , Mizuguchi Ikuyo , Akashi Tomohiro 
Published: November 2013

AbstractThe catalase gene of the pathogenic yeast Candida albicans was cloned and its expression was examined. Activity of the catalase was detected when cells which were in the early logarithmic stage were treated with hydrogen peroxide. Additionally, activity was detected without any treatment to cells in the late logarithmic and stationary phases. When cells were cultured in galactose, glycerol, or ethanol, catalase activity was always observed without the hydrogen peroxide treatment, suggesting that glucose represses the induction of catalase expression. To elucidate the molecular mechanism of catalase expression, the putative gene for catalase and its 5′ untranscribed region were cloned. Sequences of the gene and its potential regulatory region revealed several motifs, including a GC box‐like element and stress‐responsive element (STRE), which could be involved in the transcriptional regulation. Northern analysis showed that hydrogen peroxide and sorbitol activated transcription of the catalase. On the other hand, treatment of glucose strictly repressed the expression of the catalase even when co‐treated with hydrogen peroxide. The expression of catalase against treatment with hydrogen peroxide took place very quickly and decreased slowly in the experimental condition adopted here. From these results, we assumed that the expression of the catalase in Candida albicans is regulated by various environmental conditions via motifs for transcriptional activation as in other yeast catalases.
Repetitive Sequence of Leptospira interrogans Serovar Icterohaemorrhagiae Strain Ictero No. 1: A Sensitive Probe for Demonstration of Leptospira interrogans Strains
Authors: Takahashi Yukie , Kishida Masami , Yamamoto Satoru , Fukunaga Masahito 
Published: November 2013

AbstractA 4.8‐kilobase (kb) repetitive sequence element generated with KpnI digestion was cloned from the Leptospira interrogans serovar icterohaemorrhagiae strain Ictero No. 1. The sequence, repeated in tandem, was located on the 280‐kb fragment between the FseI and AscI sites on the chromosome by hybridization using the 4.8‐kb fragment as a probe. We cloned the fragment containing the element for the Ictero No. 1 strain in a lambda EMBL3 bacteriophage DNA, and one out of 5 clones was sequenced. Within the sequenced 9‐kb segment that partially repeated, 9 putative open‐reading frames and 2 transfer RNA genes, for alanine and isoleucine, were identified. A similarity search for the products deduced from the sequenced data revealed that the repeated sequence includes both beta‐oxidation enzymes, acyl‐CoA dehydrogenase and enoyl‐CoA hydratase, and hydroxythiazole kinase protein homologues. Hybridization experiments against different leptospiral strains using the element as a probe showed a similar sequence in the strains of L. interrogans and L. kirschneri, but not in any strains of L. borgpetersenii, L. weillii, L. meyeri or L. biflexa. Results indicated that the highly repeated element in the Ictero No. 1 strain exists as a well conserved sequence, though at a moderate level of repetition, in certain strains of L. interrogans and L. kirschneri. PCR amplification targeting the repetitive element was successful and indicated that the procedure provides a sensitive and specific probe to detect leptospires.
Human IgM Monoclonal Antibody to Ganglioside GM2 and Complement Suppress Virus Propagation in Ex Vivo Cultures of Lymphocytes from HIV‐1 Infected Patients
Authors: Okada Noriko , Wu Xiaoshan , Mizokami Masashi , Irie Reiko F. , Okada Hidechika 
Published: November 2013

AbstractHIV‐1 infection induces aberrant ganglioside GM2 expression on infected cell lines, and human IgM anti‐GM2 monoclonal antibody (L55 Ab) together with normal fresh human serum (FHS) as a source of complement causes complement mediated cytolysis of HIV‐1 infected cells as well as HIV‐1 particles. We report here that high expression of GM2 was also detected on HIV‐1 infected lymphocytes from HIV‐1 seropositive patients. L55 Ab effectively suppressed the generation of HIV in the presence of FHS in primarily cultured lymphocytes from HIV‐1 infected patients in ex vivo experiments, and the suppression was enhanced additively by AZT. These data suggest that L55 Ab may increase the therapeutic effect of chemotherapy.
Protective Immune Response to 16 kDa Immunoreactive Recombinant Protein Encoding the C‐Terminal VP1 Portion of Foot and Mouth Disease Virus Type Asia 1
Authors: Bayry Jagadeesh , Prabhudas Krishnamsetty , Gopalakrishna Susarla , Patil Prasanna K. , Ramakrishna Chitimalla , Misra Laxmi Dutt , Suryanarayana Veluvarti V. S. 
Published: November 2013

AbstractRecombinant protein of Foot and Mouth Disease Virus (FMDV) type Asia 1 corresponding to the C‐terminal half of VP1 was expressed in Escherichia coli. As an alternative to the synthetic peptide, this selected C‐terminal region was used as a protein vaccine in guinea pigs in order to study the immune response with various adjuvant formulations: immune stimulatory complexes (ISCOMs), Montanide ISA 206, Freund's incomplete adjuvant (FIA), lipopolysaccharide (LPS) and cytokine mixture. A primary dose of 40 μg/animal followed by a booster of the same dose was injected after a 21‐day interval. The sera were collected at intervals of 21, 42 and 63 days after the booster. The humoral response to vaccine was monitored by sandwich enzyme‐linked immunosorbent assay (ELISA) and a serum neutralization test (SNT). The guinea pig sera showed high titers both in ELISA and SNT, which could be protective. Further, irrespective of the adjuvant preparation used, the vaccine conferred protection against the challenge virus 105 days post‐vaccination in 13 of 15 animals (86%). The results indicated that a combination of recombinant protein ISCOMs and Montanide ISA 206 would be a better choice for achieving early protective titers and longer lasting immunity and that the C‐terminal half of the VP1 protein may be tried as a safe vaccine for secondary immunization.