bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prevention of Bacterial Diarrhea by Pasteurization of Drinking Water in Kenya
Authors: Iijima Yoshio , Karama Mohamed , Oundo Joseph O. , Honda Takeshi 
Published: November 2013

AbstractDiarrheal disease is one of the major causes of morbidity and mortality in developing countries. Drinking water is a primary transmission route of infectious diarrheagenic bacteria in a rural area of Kenya (Microbiol. Immunol. 41: 773–778, 1997). We tried to prevent diarrhea at villages with approximately 1,500 households in Kenya by pasteurizing drinking water. A durable simple thermoindicator which changes color at 70 C was used as an indicator of pasteurization. The number of households in which drinking water was coliform bacteria‐free increased from 10.7% to 43.1% after adoption of a pasteurization practice. Consequently, the incidence of severe diarrhea among people drinking pasteurized water was significantly lower than in people taking raw water (odds ratio = 0.55, P = 0.0016). The reduction ratio of the incidence after pasteurization was nearly equivalent with that after the adoption of a boiling method. Employment of women leaders as fieldworkers and demonstration of bacterial colony disappearance on agar plates by pasteurization also affected reduction of the diarrheal incidence.
Reconstitution and Purification of Cytolethal Distending Toxin of Actinobacillus actinomycetemcomitans
Authors: Saiki Keitarou , Konishi Kiyoshi , Gomi Tomoharu , Nishihara Tatsuji , Yoshikawa Masanosuke 
Published: November 2013

AbstractCytolethal distending toxin (CDT) has been found in various pathogenic bacterial species and causes a cell distending and a G2 arrest against eukaryotic cells. All the cdtABC genes, which encode CDT, are known to be required for the CDT activities although the CDT holotoxin structure has not been elucidated. We cloned the cdtABC genes of Actinobacillus actinomycetemcomitans and constructed an Escherichia coli expression system for them. We found that crude extracts from six deletion mutants (ΔcdtA, ΔcdtB, ΔcdtC, ΔcdtBC, ΔcdtAC, and ΔcdtAB) of recombinant E. coli, which showed very weak or no detectable CDT activities, restored the CDT activities when pre‐mixing and pre‐incubation of them were performed in combinations to contain all the CdtA, CdtB, and CdtC proteins. These results indicate that all the Cdt proteins are required for the CDT activities. We also found that the chimera CdtB protein, CdtB‐intein‐CBD (chitin binding domain) like CdtB protein itself assembled with CdtA and CdtC. The reconstituted CDT containing the chimera CdtB protein was specifically extracted by chitin beads and the only CDT portion was isolated from the chitin beads by a cleavage reaction of the intein. The purified reconstituted‐CDT was found to consist of CdtA, CdtB, and CdtC proteins, and showed appreciable CDT activities, indicating that the CDT holotoxin structure is the CdtABC complex. To our knowledge, this is the first report succeeded in complete purification of an active CDT and may offer useful tools for elucidation of the toxic mechanism of CDT.
Immunogenicity of a Recombinant MVA and a DNA Vaccine for Japanese Encephalitis Virus in Swine
Authors: Nam Jae‐hwan , Chae Soo‐lim , Cho Hae‐wol 
Published: November 2013

AbstractWe previously reported that mice immunized with recombinant modified vaccinia virus Ankara (MVA) encoding Japanese encephalitis virus (JEV) prM and E genes were completely protected against JEV challenge (Nam, J.H., Wyatt, L.S., Chae, S.L., Cho, H.W., Park, Y.K., Moss, B. Vaccine 1999, 17: 261–268). In this study, we examined the immunogenicity in swine of this recombinant MVA (vJH9) or a DNA vaccine (pcJH‐1) expressing the same JEV genes. Although the booster effect in mice with a combination of vJH9, pcJH‐1 and inactivated JEV commercial vaccine was not apparent by measuring JEV antibodies, the recombinant MVA vaccine (vJH9) and the DNA vaccine (pcJH‐1) efficiently produced neutralizing antibodies in swine and 2 doses of each showed a booster effect in mice and swine. Therefore, both vJH9 and pcJH‐1 are good candidates for a second generation JEV vaccine.
Antigenic Relatedness between Burkholderia pseudomallei and Burkholderia mallei
Authors: Anuntagool Narisara , Sirisinha Stitaya 
Published: November 2013

AbstractBurkholderia pseudomallei and Burkholderia mallei are causative agents of distinct diseases, namely, melioidosis and glanders, respectively. The two species are very closely related, based on DNA‐DNA homology, base sequence of the 16S rRNA, and phenotypic characteristics. Based on the use of polyclonal antisera, B. pseudomallei and B. mallei are also found to be antigenically closely related to one another. We previously reported the production of monoclonal antibodies (MAbs) against B. pseudomallei antigens; one group was specific for the 200‐kDa exopolysaccharide present on the surface of all B. pseudomallei isolates, and the other was specific for the lipopolysaccharide (LPS) structure present on more than 95% of the B. pseudomallei tested. In the present study, we showed that the MAbs against 200‐kDa antigen of B. pseudomallei cross‐reacted with a component present also in some B. mallei isolates (3/6), but the positive immunoblot reaction was noted below the 200‐kDa position. On the other hand, none of the six B. mallei isolates reacted with the MAb specific for B. pseudomallei LPS. It was of interest to observe that only the 3 exopolysaccharide‐positive B. mallei isolates reacted with a commercial MAb against B. mallei LPS. The data presented suggest that B. mallei can be classified antigenically into two types based on their reactivities with different MAbs, i.e., the presence or absence of exopolysaccharide and the types of lipopolysaccharide. The heterogeneity of the LPS from these two closely related organisms is most likely related to the differences in its O‐polysaccharide side chain.
Heat Stability of Carboxypeptidase R of Experimental Animals
Authors: Komura Hidefumi , Shimomura Yasuyo , Yumoto Miho , Katsuya Hirotada , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractCarboxypeptidase N (CPN) and carboxypeptidase R (CPR) are present in fresh serum, and cleave C‐terminal arginine or lysine residues from bioactive peptides such as anaphylatoxins and kinins resulting in regulation of peptide activity. Although CPN is present in the active form in plasma, CPR is generated from proCPR by trypsin‐like enzymes such as thrombin. CPR regulates not only inflammatory peptides but also restricts fibrinolysis. To elucidate the complex role of CPN and CPR in vivo, studies in animal models will be essential. CPR of guinea pig, rat and rabbit decayed at 37 C rapidly as in the case of human CPR. However, at 25 C, CPR of those species decayed to some extent, although human serum CPR did not decay within 60 min. In the presence of thrombin inhibitor, CPR in the sera of animals tested decayed more rapidly than CPR in serum without thrombin inhibitor suggesting that additional generation of CPR may have been prevented during decay evaluation. However, human serum CPR decayed more rapidly in the absence of thrombin inhibitor indicating that thrombin may accelerate the decay in human serum.
An Outbreak of Neonatal Toxic Shock Syndrome‐Like Exanthematous Disease (NTED) Caused by Methicillin‐Resistant Staphylococcus aureus (MRSA) in a Neonatal Intensive Care Unit
Authors: Nakano Miyo , Miyazawa Hirofumi , Kawano Yasushi , Kawagishi Mika , Torii Keizo , Hasegawa Tadao , Iinuma Yoshitsugu , Ohta Michio 
Published: November 2013

AbstractNeonatal toxic shock syndrome‐like exanthematous disease (NTED) is a new entity of methicillin‐resistant Staphylococcus aureus (MRSA) infection. Most of NTED cases reported previously in the literature were sporadic ones. In the present report, we describe an outbreak of NTED that occurred in a neonatal intensive care unit (NICU) between April, 1999 and April, 2000 in Japan. All MRSA strains isolated from 14 patients (6 NTED, 2 infections and 6 colonizations) in this outbreak belonged to the group of coagulase II and produced toxic shock syndrome toxin 1 (TSST‐1). Of these, 14 strains produced staphylococcal enterotoxin C (SEC). No other superantigenic toxins were produced by these strains. The pulsed field gel electrophoresis (PFGE) patterns of genomic DNA digested with SmaI were indistinguishable each other due to no band shifting in all of the 13 strains except for strain O‐21 and M56. Strain M56 was different from the dominant type in the positions of only 2 bands, whereas the pattern of strain O‐21 had no similarity with the other pattern, suggesting that this outbreak was associated with the spread of a unique MRSA strain in the NICU. Two‐dimensional electrophoresis (2‐DE) analysis of exoproteins revealed that the patterns of these 14 strains were very indistinguishable to each other, and that these strains produced very large amounts of TSST‐1 and SEC3 subtype superantigens, as measured with computer‐assisted image analysis of the intensity of 2‐DE spots. The 2‐DE gel of O‐21 showed the different pattern from the others. These results as well as the profiles of toxin production also supported the conclusion drawn from PFGE analysis. Based on these results, the involvement of TSST‐1 and SEC3 in the pathogenesis of NTED is discussed.
Rapid and Sensitive Detection Method of a Bacterium by Using a GFP Reporter Phage
Authors: Funatsu Takashi , Taniyama Tadayoshi , Tajima Takashi , Tadakuma Hisashi , Namiki Hideo 
Published: November 2013

AbstractA rapid, sensitive, and convenient method for detecting a specific bacterium was developed by using a GFP phage. Here we describe a model system that utilizes the temperate Escherichia coli‐restricted bacteriophage lambda, which was genetically modified to express a reporter gene for GFP to identify the colon bacillus E. coli in the specimen. E. coli infected with GFP phage was detected by GFP fluorescence after 4–6 hr of incubation. The results show that a few bacteria in a specimen can be detected under fluorescence microscopy equipped with a sensitive cooled CCD camera. When E. coli and Mycobacterium smegmatis were mixed in a solution containing GFP phage, only E. coli was infected, indicating the specificity of this method. The method has the following advantages: 1) Bacteria from biological samples need not be purified unless they contain fluorescent impurities; 2) The infection of GFP phage to bacteria is specific; 3) The fluorescence of GFP within infected bacteria enables highly sensitive detection; 4) Exogenous substrates and cofactors are not required for fluorescence. Therefore this method is suitable for any phage‐bacterium system when bacteria‐specific phages are available.
Evaluation of Therapeutic Efficacy of Adjuvant Helicobacter pylori Whole Cell Sonicate in Mice with Chronic H. pylori Infection
Authors: Maeda Kosaku , Yamashiro Tetsu , Minoura Takanori , Fujioka Toshio , Nasu Masaru , Nishizono Akira 
Published: November 2013

AbstractSuccessful prophylactic administration of Helicobacter pylori whole cell sonicate (WCS) plus complete Freund's adjuvant (CFA) or aluminum hydroxide (ALM) against subsequent H. pylori infection was reported recently. Here we tested the effect of WCS plus TiterMax® Gold (TMX) or ALM in mice with chronic H. pylori infection. Mice with chronic (18 weeks) H. pylori infection were injected intraperitoneally with H. pylori (Sydney strain) WCS plus ALM or TMX once weekly for three times. The number of colonizing H. pylori in the stomach, IgG1 and IgG2a levels, and local inflammatory status were determined after therapeutic immunization. H. pylori specific IgG1, but not IgG2a, was significantly induced in mice immunized with H. pylori WCS plus TMX or ALM. Immunization did not result in reduction of bacterial count or recruiting inflammatory cells to the stomach. Adjuvant H. pylori WCS resulted in induction of CD4+ Th2 cell‐mediated immunity although it did not reduce bacterial density in mice with chronic H. pylori infection. Our results implied that CD4+ Th1 cell‐mediated immunity, rather than Th2 cell dominant immunity, might play a role in reducing the number of bacteria in chronic H. pylori infection.
A Very Late Activating Antigen‐α4 (CD49d) Monoclonal Antibody, BU49 Induces Phosphorylation of a cAMP Response Element‐Binding Protein (CREB), Resulting in Induction of Homotypic Cell Aggregation and Enhancement of Interleukin‐8 (IL‐8) Production
Authors: Ikewaki Nobunao , Inoko Hidetoshi 
Published: November 2013

AbstractA very late activating antigen‐α4 (CD49d) monoclonal antibody (mAb), BU49 was found to induce phosphorylation of a cAMP response element‐binding protein (CREB) in the human monocyte‐like cell line, U937. This phosphorylation of CREB was completely inhibited by a protein kinase A (PKA) inhibitor H‐89 with the optimum concentration (completely inhibits PKA). Furthermore, BU49 strongly and rapidly (within 5 hr) induced homotypic cell aggregation in the U937 cells accompanied by CREB phosphorylation. This cell aggregation was also completely inhibited by the addition of H‐89. Interestingly, both of two mAbs (mAb13 and 4B4) recognizing different epitopes on the CD29 (β1 integrin) completely inhibited this aggregation at the late phase (18 to 24 hr) but not at the early phase (5 hr) after cultured with BU49. On the other hand, BU49 significantly enhanced interleukin‐8 (IL‐8) production from the U937 cells into the culture supernatant. In addition, this IL‐8 production was significantly blocked in the presence of H‐89 with the optimum concentration. However, a CD29 mAb which inhibits homotypic cell aggregation could not block this IL‐8 production. Taken together, these findings indicate that BU49 induces CREB phosphorylation mainly mediated by PKA, which finally results in the induction of homotypic cell aggregation and the enhancement of IL‐8 production. Furthermore, these findings also indicate that the enhancement of IL‐8 production from the U937 cells induced by BU49 partially depends on CREB phosphorylation mainly mediated by PKA.
Role of Internalization in the Pathogenicity of Shiga Toxin‐Producing Escherichia coli Infection in a Gnotobiotic Murine Model
Authors: Aiba Yuji , Ishikawa Hiroki , Shimizu Keiko , Noda Satoshi , Kitada Yukie , Sasaki Masafumi , Koga Yasuhiro 
Published: November 2013

AbstractWe investigated the role of bacterial internalization in the killing caused by Shiga toxin‐producing Escherichia coli (STEC) infection using a gnotobiotic murine model. A high number of internalized STEC was found in the colonic epithelial cells of STEC‐infected mice by both an ex vivo assay and transmission electron microscopy. Most of these mice were killed within 10 days after infection. However, the implantation of lactic acid bacteria in such mice before infection markedly decreased the number of internalized STECs and also completely protected these hosts from killing by a STEC infection. The inhibition of such internalization by immunoglobulin also prevented the hosts from being killed. The Shiga toxin levels in these hosts indicated an inhibition of the penetration of Shiga toxins produced in the colon to the underlying tissue. These results suggested that the internalization plays an important role in the pathogenicity caused by STEC infection in a gnotobiotic murine model.
Lethal Factor to Mice Produced by Escherichia coli Isolated from Chickens with Swollen Head Syndrome
Authors: Salvadori Marcia Regina , Chudzinski‐Tavassi Ana Marisa , Baccaro Maria Regina , Ferreira Claudete Serrano Astolfi , Ferreira Antonio José Piantino , Prado‐Franceschi Júlia , Yano Tomomasa 
Published: November 2013

AbstractA lethal toxin similar to Bacillus cererus lethal toxin was detected in the culture supernatants of Escherichia coli isolated from chickens with swollen head syndrome. The lethal activity was heat‐labile, protease‐sensitive and killed mice within 10 min. The light microscopy of the histopathological studies revealed that the principal organ affected by this toxin was the lung but the liver and kidneys also showed lesions. The relevance of this lethal activity from E. coli remains to be determined.
Identification of the Pathogenic Aspergillus Species by Nested PCR Using a Mixture of Specific Primers to DNA Topoisomerase II Gene
Authors: Kanbe Toshio , Yamaki Kenichi , Kikuchi Akihiko 
Published: November 2013

AbstractFor PCR‐based identification of Aspergillus species, a common primer of the DNA topoisomerase II genes of Candida, Aspergillus and Penicillium, and species‐specific primers of the genomic sequences of DNA topoisomerase II of A. fumigatus, A. niger, A. flavus (A. oryzae), A. nidulans and A. terreus were tested for their specificities in PCR amplifications. The method consisted of amplification of the genomic DNA topoisomerase II gene by a common primer set, followed by a second PCR with a primer mix consisting of 5 species‐specific primer pairs for each Aspergillus species. By using the common primer pair, a DNA fragment of approximately 1,200 bp was amplified from the Aspergillus and Penicillium genomic DNAs. Using each species‐specific primer pair, unique sizes of PCR products were amplified, all of which corresponded to a species of Aspergillus even in the presence of DNAs of several fungal species. The sensitivity of A. fumigatus to the nested PCR was found to be 100 fg of DNA in the reaction mixture. In the nested PCR obtained by using the primer mix (PsIV), the specific DNA fragment of A. fumigatus was amplified from clinical specimens. These results suggest that this nested PCR method is rapid, simple and available as a tool for identification of pathogenic Aspergillus to a species level.
Construction and In Vitro Properties of Chimeric Simian and Human Immunodeficiency Virus with the Human TNF‐Alpha Gene
Authors: Haga Takeshi , Shimizu Yuya , Okoba Masashi , Kumabe Shino , Goto Yoshitaka , Shinjo Toshiharu , Ichimura Hiroshi , Kuwata Takeo , Hayami Masanori , Miura Tomoyuki 
Published: November 2013

AbstractTumor necrosis factor‐alpha (TNF‐α) has been reported to be involved in the development and progression of acquired immunodeficiency syndrome (AIDS). To study the role of this cytokine in AIDS pathogenesis, we constructed a chimeric simian and human immunodeficiency virus (SHIV) having the human TNF‐α gene (SHIV‐TNF) and characterized its properties in vitro. SHIV‐TNF replicated both in M8166, a human T cell line, and in monkey peripheral blood mononuclear cells (PBMCs). Along with SHIV‐TNF replication, TNF‐α was detected in the culture supernatant by ELISA. The maximum expression level of TNF‐α reached 120 ng/ml in M8166 cells, and 2.5 ng/ml in monkey PBMCs. The expressed TNF was biologically active, as shown by a cytotoxic assay using TNF‐sensitive L929 mouse fibroblasts. This activity was detected at least until 10 passages of SHIV‐TNF (74 days after the initial infection). In monkey PBMCs, SHIV‐TNF replicated much better than the parental SHIV‐NI. Flow cytometric analysis showed that the death of monkey PBMCs infected with SHIV‐TNF was severer than that caused by the parental SHIV‐NI. These results suggest that SHIV‐TNF would be useful for inducing the disease in a monkey model, which may contribute to a better understanding of the role of TNF‐α in AIDS etiology.
Serum Antibodies of Periodontitis Patients Compared to the Lipopolysaccharides of Porphyromonas gingivalis and Fusobacterium nucleatum
Authors: Onoue Sakura , Imai Toshio , Kumada Hidefumi , Umemoto Toshio , Kaca Wieslaw , Isshiki Yasunori , Kaneko Aki , Kawahara Kazuyoshi 
Published: November 2013

AbstractSerum antibody titers against the lipopolysaccharides (LPSs) of Porphyromonas gingivalis and Fusobacterium nucleatum were compared between 9 periodontitis patients and 24 healthy persons. The IgG titers against the LPSs of P. gingivalis ATCC 33277T and W50 were clearly higher in the patients than in the healthy persons. However, IgM titers against the LPSs of P. gingivalis strains were relatively low, and no significant difference was observed between the patients and healthy persons. On the other hand, IgG and IgM titers against the LPS of Fusobacterium nucleatum JCM 8532T in some patients were significantly higher than those in the healthy persons, although the difference in IgG titers was not large compared to that of the LPS of P. gingivalis. These results suggest that the antibody measurement of patients' sera against the LPS of periodontal bacteria can be applied for the diagnosis of periodontitis.
Tyrosine Phosphorylation of a Novel 100‐kDa Protein Coupled to CD28 in Resting Human T Cells Is Enhanced by a Signal through TCR/CD3 Complex
Authors: Matsumoto Aki , Dobashi Hiroaki , Ohnishi Hiroaki , Tanaka Terukazu , Kubota Yoshitsugu , Kitanaka Akira , Ishida Hiroshi , Tokuda Michiaki , Waki Masato , Kubo Akihito , Ishida Toshihiko 
Published: November 2013

AbstractFor T cell activation, two signals are required, i.e., a T cell receptor (TCR)/CD3‐mediated main signal and a CD28‐mediated costimulatory signal. CD28 binds to its ligand (CD80 or CD86) and transduces the most important costimulatory signal. The cytoplasmic domain of the CD28 molecule, composed of 41 amino acids, does not contain any intrinsic enzyme activity. The cytoplasmic domain of CD28 is remarkably conserved among species and is associated with a number of signaling molecules that affect the main signal. We report here that a tyrosine phosphorylated 100‐kDa protein (pp100) was coupled to the CD28 cytoplasmic domain in Jurkat and human peripheral T cells. The pp100 was distinguished from other CD28 associated molecules such as Vav, STAT5, PI 3‐kinase, Valosin‐containing protein (VCP), Nucleolin, Gab2 (Grb2‐associated binding protein 2), and STAT6. The tyrosine phosphorylation of pp100 coprecipitated with CD28 was enhanced by CD3 stimulation by the specific antibody, tyrosine phosphatase inhibitor and PKC activator. Tyrosine phosphorylation of pp100 was attenuated by the prior addition of PKC inhibitor. These findings indicate that pp100 is a novel tyrosine phosphorylated protein coupled to CD28 under continuous control of tyrosine phosphatases and might play a role in T cell activation augmented by a TCR/CD3‐mediated main signal.
High Molecular Weight Factor in FCS Inhibits Helicobacter pylori VacA‐Binding to Its Receptor, RPTPβ, on AZ‐521
Authors: Kimura Takahiro , Wada Akihiro , Nakayama Masaaki , Ogushi Ken‐ichi , Nishi Yoshito , De Guzman Blanquita B. , Moss Joel , Hirayama Toshiya 
Published: November 2013

AbstractVacA, a secretory product of Helicobacter pylori, binds to its cell surface receptor, receptor tyrosine phosphatase (RPTP) β, leading to cytoplasmic vacuolization of gastric epithelial AZ‐521 cells. VacA binding to the cell surface and VacA‐dependent vacuolization were inhibited by cell culture media containing fetal calf serum (FCS). The high molecular weight fraction of FCS isolated by Superose 12 gel filtration chromatography inhibited VacA binding, whereas only weak effects were observed with other fractions. These data show that the high molecular weight fraction of FCS inhibits VacA action though its ability to block toxin binding to its receptor, RPTPβ, on AZ‐521 cells.