bluuebunny/crossref_metadata_2025_split huggingface dataset:
Infection with Campylobacter jejuni Induces Tyrosine‐Phosphorylated Proteins into INT‐407 Cells
Authors: Biswas Debabrata , Niwa Hidekazu , Itoh Kikuji 
Published: November 2013

AbstractThe mechanisms used by Campylobacter jejuni to induce internalization into host intestinal epithelial cells have not been defined. In this study, we obtained evidence that exposure of INT‐407 cells to protein kinase inhibitors results in decreased invasion of these cells by C. jejuni in a dose dependent manner. Preincubation of INT‐407 cells in the presence of staurosporine, tyrphostin 46 and genistein decreased invasion of these cells by C. jejuni significantly. Moreover, C. jejuni infection of INT‐407 cells induced tyrosine phosphorylation of several Triton X‐100 soluble proteins with approximate molecular weights of 170, 145, 90, 60 and 55 kDa that were absent or reduced in the presence of genistein in cells after 1 hr of pretreatment. These data suggest that tyrosine protein kinase‐linked pathways strongly regulate the internalization of C. jejuni into intestinal epithelial cells.
Identification of Signature Genes for Rapid and Specific Characterization of Yersinia pestis
Authors: Zhou Dongsheng , Han Yanping , Dai Erhei , Pei Decui , Song Yajun , Zhai Junhui , Du Zongmin , Wang Jin , Guo Zhaobiao , Yang Ruifu 
Published: November 2013

AbstractPolymerase chain reaction (PCR) amplification of DNA‐based unique markers, the signature sequences, is ideal for rapid detection and identification of pathogens. We described the discovery of twenty‐eight signature genes of Yersinia pestis by DNA microarray‐based comparative genome hybridization in conjunction with PCR validation. Three pairs of Y. pestis‐specific primers designed from signature genes were demonstrated to have the expected specificity to this target bacterium, without cross‐reaction with the closely related Y. pseudotuberculosis or a large collection of genomic DNAs from other organisms.
Effects of Recombinant Cholera Toxin B Subunit (rCTB) on Cellular Immune Responses: Enhancement of Delayed‐Type Hypersensitivity Following Intranasal Co‐Administration of Mycobacterium bovis‐BCG with rCTB
Authors: Maeyama Jun‐ichi , Isaka Masanori , Yasuda Yoko , Matano Keiko , Morokuma Kazunori , Ohkuma Kunio , Tochikubo Kunio , Yamamoto Saburo , Goto Norihisa 
Published: November 2013

AbstractRecombinant cholera toxin B subunit (rCTB) is a safe and potent mucosal adjuvant. To gain insight into the mechanism underlying the adjuvant effect of rCTB, the effects of rCTB on cell‐mediated immune responses of mice and guinea pigs were examined after intranasal administration of Mycobacterium bovis‐bacillus Calmette‐Guérin (BCG) with and without rCTB. Delayed‐type hypersensitivity, for skin reactions in guinea pigs and for footpad swelling reactions in mice, to purified protein derivative (PPD) were enhanced by intranasal co‐administration of BCG and rCTB, as compared to giving BCG alone to these animals. Moreover, tumor necrosis factor (TNF)‐α and interferon (IFN)‐γ production of spleen cells and antigen specific spleen cell proliferation, stimulated with PPD, were enhanced in the presence of rCTB. These results strongly suggest that rCTB enhances cellular as well as humoral immune responses.
Zymographic Characterization of Bacteriolytic Enzymes Produced by Oral Streptococci
Authors: Yoshimura Goh , Komatsuzawa Hitoshi , Kajimura Junko , Fujiwara Tamaki , Ohara Masaru , Kozai Katsuyuki , Sugai Motoyuki 
Published: November 2013

AbstractZymographic analysis was performed to know the bacteriolytic enzyme profiles of 4% SDS extracts of oral streptococci, Streptococcus mutans, S. sobrinus, S. sanguis, S. mitis and S. salivarius. We investigated the five strains in each species and found that the profile was very similar among strains of the same species except for S. salivarius (the profile was classified into two types). On the other hand, the profile was considerably different among species. Two major bacteriolytic enzymes of S. mutans showing molecular mass of 80 and 100 kDa were found using SDS‐boiled S. mutans or S. sobrinus cells as substrate. These bacteriolytic activities were less apparent in the gel containing S. mitis or S. salivarius, and also not detectable in the gel containing S. sanguis. S. sobrinus extract showed only one bacteriolytic band (78 kDa) as strong activity using S. sobrinus cells as substrate. S. sanguis extract showed no bacteriolytic bands using any streptococcal cells. Extracts of either S. mitis or S. salivarius showed weak activity by using respective strains as substrate.
Epidemiological Characterization of Salmonella Typhimurium DT104 Prevalent among Food‐Producing Animals in the Japanese Veterinary Antimicrobial Resistance Monitoring Program (1999–2001)
Authors: Esaki Hidetake , Morioka Ayako , Kojima Akemi , Ishihara Kanako , Asai Tetsuo , Tamura Yutaka , Izumiya Hidemasa , Terajima Jyun , Watanabe Haruo , Takahashi Toshio 
Published: November 2013

AbstractIn the course of nationwide investigation on epidemiological characteristics in Salmonella Typhimurium isolates from food‐producing animals in Japan between 1999 and 2001, fifty‐seven isolates of S. Typhimurium DT104 and 104B obtained from cattle and swine at farm level in Japan between 1999 and 2001 were classified with pulsotype and antimicrobial resistance type. Most of the isolates were resistant to five or more antimicrobials and were genotyped into four groups. The present nationwide investigation shows that at least 11 types of S. Typhimurium related to DT104 are prevalent among food‐producing animals across the country.
Detection of the Impairment of CD80 Expression on Circulating Monocytes in HIV‐Infected Thai Children
Authors: Ubol Sukathida , Aiamkitsumrit Benjamaz , Nitayaphan Sorachai , Sirinavin Sayomporn , Kitthawee Sangvorn 
Published: November 2013

AbstractThe mechanism of progressive anergic response in HIV‐infected children has yet to be adequately described. One possibility is inappropriate delivery of an essential second signal for T‐cell activation due to the inappropriate presentation of co‐stimulatory molecules. To determine whether the ligand for the secondary signal is impaired in pediatric AIDS, we compared the level of CD80 expression by circulating monocytes in HIV‐infected and ‐noninfected children (15 mild/asymptomatic, 13 symptomatic and 12 HIV seronegative children). By two‐color flow cytometry analysis, there was no statistically significant difference in the percentage of monocytes expressing CD80 among the groups (i.e., 63.2±15.8, 60.9±12.7, 61.04±10.9 for uninfected children, mild‐asymptomatic children and symptomatic children, respectively). However, both infected groups showed statistically significant lower levels of CD80 expression, with mean fluorescent intensities of 40.9±15.9 and 38.8±10.7 compared to 57.05±16.3 for the uninfected control group. Our data demonstrated a correlation between HIV infection and impairment of CD80 by circulating monocytes. Whether the impairment on CD80 expression contributes to destruction of the immunological network in HIV‐infected children requires further investigation.
Microbiota Composition of the Intestinal Mucosa: Association with Fecal Microbiota?
Authors: Ouwehand Arthur C. , Salminen Seppo , Arvola Taina , Ruuska Tarja , Isolauri Erika 
Published: November 2013

AbstractThe fecal and mucosal microbiota of infants with rectal bleeding and the fecal microbiota of healthy age‐matched controls were investigated by fluorescent in situ hybridization. Bifidobacteria were the main genus in both the feces and mucosa. The other genera tested, Bacteroides, Clostridium, Escherichia coli and lactobacilli/enterococci, represented only minor constituents. No differences in fecal microbiota were observed between patients and controls. In the patients, however, four times greater numbers of bifidobacteria were observed in the feces when compared to the mucosa. Notwithstanding this difference, a strong positive correlation prevailed for bifidobacteria in feces and mucosal samples. The genera assessed accounted for 16% of total bacterial counts on mucosal samples and for 47% of total bacterial counts in feces. This indicates that the unidentified part of the microbiota, especially on the mucosa, deserves more attention.
Epidemiological and Serological Survey of Brucellosis in Mongolia by ELISA Using Sarcosine Extracts
Authors: Erdenebaatar Janchivdorj , Bayarsaikhan Balgan , Yondondorj Agchbazar , Watarai Masahisa , Shirahata Toshikazu , Jargalsaikhan Enkhtuya , Kawamoto Keiko , Makino Sou‐ichi 
Published: November 2013

AbstractBrucellosis is an important zoonosis, and serological surveillance is essential to its control. However, cross‐reactions of attenuated live cells of Brucella abortus strain S‐19 and B. melitensis strain Rev‐1 with Yersinia enterocolitica O9 or vaccinated animal sera interfere with accurate serological diagnosis by the Rose Bengal test (RBT). Therefore, we used ELISA with sarcosine extracts from the virulent B. abortus strain 544 to eliminate false‐positives among RBT positive‐sera. A total of 697 serum samples were collected in Mongolia from humans and animals in 23 nomadic herds. The herds were classified into three groups as brucellosis‐endemic (BE), brucellosis‐suspected (BS), or Brucella‐vaccinated (BV). The number of 295 animals (43.0%) was positive by RBT, but 206 (69.8%) of these were positive according to ELISA; therefore, 30.2% of the RBT‐positive sera were found to be false positives. The false positive samples for RTB represent 4.1%, 27.4%, and 68.2% of the animals from the BE, BS, and BV herds, respectively. In addition, 32% of RBT‐positive human sera were also false positives. Thus, our ELISA would be more specific than RTB and useful for epidemiological surveillance for brucellosis.
Suppression of Hepatitis C Virus Replicon by RNA Interference Directed against the NS3 and NS5B Regions of the Viral Genome
Authors: Takigawa Yuki , Nagano‐Fujii Motoko , Deng Lin , Hidajat Rachmat , Tanaka Motofumi , Mizuta Hiroyuki , Hotta Hak 
Published: November 2013

AbstractRNA interference (RNAi) is a phenomenon in which small interfering RNA (siRNA), an RNA duplex 21 to 23 nucleotides (nt) long, or short hairpin RNA (shRNA) resembling siRNA, mediates degradation of the target RNA molecule in a sequence‐specific manner. RNAi is now expected to be a useful therapeutic strategy for hepatitis C virus (HCV) infection. In the present study we compared the efficacy of a number of shRNAs directed against different target regions of the HCV genome, such as 5′‐untranslated region (5′UTR) (nt 286 to 304), Core (nt 371 to 389), NS3–1 (nt 2052 to 2060), NS3–2 (nt 2104 to 2122), and NS5B (nt 7326 to 7344), all of which except for NS5B are conserved among most, if not all, HCV subtype 1b (HCV‐1b) isolates in Japan. We utilized two methods to express shRNAs, one utilizing an expression plasmid (pAVU6+27) and the other utilizing a recombinant lentivirus harboring the pAVU6+27‐derived expression cassette. Although 5′UTR has been considered to be the most suitable region for therapeutic siRNA and/or shRNA because of its extremely high degree of sequence conservation, we observed only a faint suppression of an HCV subgenomic replicon by shRNA against 5′UTR. In both plasmid‐ and lentivirus‐mediated expression systems, shRNAs against NS3–1 and NS5B suppressed most efficiently the replication of the HCV replicon without suppressing host cellular gene expression. Synthetic siRNA against NS3–1 also inhibited replication of the HCV replicon in a dose‐dependent manner. Taken together, the present results imply the possibility that the recombinant lentivirus expressing shRNA against NS3–1 would be a useful tool to inhibit HCV‐1b infection.
A Highly Effective Method for Removing Suspended Poliovirus from Water Using a Positively‐Charged Carbon Felt Electrode
Authors: Kondo Yasuhito , Morita Yukio , Yamada Atsushi , Kimura Hirokazu 
Published: November 2013

AbstractWe developed an effective system to eliminate poliovirus from modified tap water using a positively‐charged carbon felt electrode. The zeta potential of polioviruses was measured using laser microscopic electrophoresis. Poliovirus adsorption to the electrode was examined by indirect immunofluorescence. The tissue culture infective dose (TCID) of poliovirus type 2 (Sabin strain) was determined using human rhabdomyosarcoma cells (RD cells). Poliovirus VP2 gene copy numbers were assessed by reverse transcription followed by a quantitative real‐time polymerase chain reaction. The mean zeta potential of the viruses was −20 mV. Relatively large numbers of polioviruses (103 or 4 × 103 TCID50/0.1 ml) could be removed by adsorption to the electrode, drastically decreasing TCID and copy numbers of poliovirus genome in the water. Virus elimination was dependent on electric current and time. Thus, the positively‐charged carbon felt electrode effectively adsorbed polioviruses. The system may prove applicable to the elimination of certain viruses from water.
PCR‐Based Identification of Pandemic Group Vibrio parahaemolyticus with a Novel Group‐Specific Primer Pair
Authors: Okura Masatoshi , Osawa Ro , Iguchi Atsushi , Takagi Michihiro , Arakawa Eiji , Terajima Jun , Watanabe Haruo 
Published: November 2013

AbstractA PCR‐based assay to identify pandemic group Vibrio parahaemolyticus has been developed. The assay employs an oligonucleotide primer pair derived from the group‐specific sequence of an arbitrarily primed‐PCR fragment, which is located in the genome encoding a “hypothetical protein,” approximately 80% homologous to the Mn2+ and Fe2+ transporter of the NRAMP family of V. vulnificus. The assay distinguished the pandemic group from other V. parahaemolyticus strains by yielding a 235‐bp specific amplicon, and can be a useful diagnostic tool for identification of pandemic group strains.
Piceatannol Prevents Lipopolysaccharide (LPS)‐Induced Nitric Oxide (NO) Production and Nuclear Factor (NF)‐κB Activation by Inhibiting IκB Kinase (IKK)
Authors: Islam Shamima , Hassan Ferdaus , Mu Mya Mya , Ito Hiroyasu , Koide Naoki , Mori Isamu , Yoshida Tomoaki , Yokochi Takashi 
Published: November 2013

AbstractThe effect of piceatannol on lipopolysaccharide (LPS)‐induced nitric oxide (NO) production was examined. Piceatannol significantly inhibited NO production in LPS‐stimulated RAW 264.7 cells. The inhibition was due to the reduced expression of an inducible isoform of NO synthase (iNOS). The inhibitory effect of piceatannol was mediated by down‐regulation of LPS‐induced nuclear factor (NF)‐κB activation, but not by its cytotoxic action. Piceatannol inhibited IκB kinase (IKK)‐α and β phosphorylation, and subsequently IKB‐α phosphorylation in LPS‐stimulated RAW 264.7 cells. On the other hand, piceatannol did not affect activation of mitogen‐activated protein (MAP) kinases including extracellular signal regulated kinase 1/2 (Erk1/2), p38 and stress‐activated protein kinase/c‐Jun NH2‐terminal kinase (SAPK/JNK). Piceatannol inhibited the phosphorylation of Akt and Raf‐1 molecules, which regulated the activation of IKK‐α and β phosphorylation. The detailed mechanism of the inhibition of LPS‐induced NO production by piceatannol is discussed.
Comparative Evaluation of Standard Dilution Method and Commercial Kit for Frozen Plate Antifungal Susceptibility Testing of Yeasts Using 200 Clinical Isolates
Authors: Makimura Koichi , Suzuki Takako , Tamura Takashi , Ikedo Masanari , Hanazawa Ryo , Takahashi Yuko , Yamada Yohko , Uchida Katsuhisa , Yamaguchi Hideyo 
Published: November 2013

AbstractA comparative evaluation of standard microdilution methods and a commercial kit for frozen plate antifungal susceptibility testing of yeasts was performed using amphotericin B, flucytosine, fluconazole, miconazole, and itraconazole on 200 yeast isolates. The isolates included 100 strains of Candida albicans, eight of C. tropicalis, twelve of C. parapsilosis, eight of C. glabrata, five of Cryptococcus neoformans, thirteen of Trichosporon asahii, and 54 other strains of seven other species of ascomycotic yeasts. Microdilution testing was performed according to the standard method for antifungal susceptibility testing published by the Japanese Society for Medical Mycology (JSMM), which are a modification of the method developed by the National Committee for Clinical Laboratory Standards (NCCLS) M27‐P. The commercial kit was prepared according to the manufacturer's instructions. The degree of agreement within ± 1 dilution for 200 clinical isolates against five antifungal agents was excellent with values for amphotericin B, flucytosine, fluconazole, miconazole, and itraconazole of 100%, 99.0%, 97.5%, 97.0%, and 97.0%, respectively. Overall, the frozen plate antifungal susceptibility testing kit provided convenient and reproducible results comparable to those obtained with the JSMM standard method.
Delayed HIV‐1 Infection of CD4+ T Lymphocytes from Therapy‐Naïve Patients Demonstrated by Quantification of HIV‐1 DNA Copy Numbers
Authors: Wada Kaoru , Nagai Hiromi , Hagiwara Tomoko , Ibe Shiro , Utsumi Makoto , Kaneda Tsuguhiro 
Published: November 2013

AbstractMeasuring the amount of HIV‐1 DNA in infected cells is important to estimate the size of the viral reservoir in patients. However, the clinical impact of the intracellular viral DNA level remains unclear. The present study examines the clinical significance of the HIV‐1 DNA level in peripheral CD4+ T lymphocytes from 21 therapy‐naïve patients. HIV‐1 DNA levels in purified peripheral CD4+ T lymphocytes were measured by the real‐time PCR method using the Roche LightCycler system that can detect 200 copies/106 cells. We detected intracellular HIV‐1 DNA in 15 (71.4%) of 21 patients at levels ranging from 270 to 98,120 copies/106 CD4+ cells, with a median of 2,220 copies/106 cells. We also found HIV‐1 DNA that was below the detection limit in the remaining 6 patients, although 8,800–150,000 copies/ml of HIV‐1 RNA were detected in plasma. Circular HIV‐1 DNA was not detected in 5 of 6 cases, suggesting that reverse transcription in CD4+ T lymphocytes of these cases was not active. Thus, delayed HIV‐1 infection of CD4+ T lymphocytes was demonstrated in these patients. The level of HIV‐1 DNA in peripheral CD4+ T lymphocytes indicates the clinical status of therapy‐naïve patients.
Study on Immunological Status of Chinese HIV‐Infected Individuals
Authors: Shang Hong , Jiang Yongjun , Zhang Zining , Han Xiaoxu , Zhang Min , Wang Yanan , Liu Jing 
Published: November 2013

AbstractHIV‐1 infection is characterized by a gradual decline of immune function, and the immune dysfunction is widely regarded as one of the most important determinants of disease progression. The present study was performed to analyze in more detail the immunological status of HIV‐infected people in China. T cell counts, activation of T cells, HIV‐1 specific CTL and plasma levels of cytokines were determined with flow cytometry, IFN‐γ Elispot or ELISA techniques. The HIV viral load was negatively correlated with CD4+, CD8+ T cell counts (r = –0.654, P<0.001; r = –0.228, P<0.05); the breadth and magnitude of HIV‐1 specific CTL responses against HIV‐1 Gag peptides was related to disease progression; the activation of CD8+ T cells was significantly higher than that in HIV‐negative controls; the level of plasma IL‐12 was much lower and the plasma IFN‐γ, IL‐10 and IL‐6 were much higher in HIV‐infected persons than in HIV‐negative controls (P<0.05). Study on immunological status in HIV‐infected Chinese is very important in predicting the disease progression and providing information for HAART therapy in China.
Novel Evidence Suggesting Clostridium difficile Is Present in Human Gut Microbiota More Frequently than Previously Suspected
Authors: Iizuka Masahiro , Konno Shiho , Itou Hiroaki , Chihara Junichi , Toyoshima Itaru , Horie Yasuo , Sasaki Kenji , Sato Akiko , Shindo Kenichi , Watanabe Sumio 
Published: November 2013

AbstractPrevalence rate of Clostridium difficile in healthy human adults is believed to be very low. Our RT‐PCR system using glass powder, which can eliminate PCR inhibitors, detected C. difficile toxin B mRNA in 16 of 30 fecal samples (53.3%) from healthy human adults. In contrast, we failed to detect toxin B in the same fecal samples by PCR using DNA templates extracted with phenol‐chloroform. Our results suggest that PCR inhibitors in feces carried through phenol‐chloroform extraction procedure might suppress the sensitivity of PCR and that C. difficile is actually present in human gut microbiota more frequently than previously suspected.
Effects of Two Probiotic Lactobacillus Strains on Jejunal and Cecal Microbiota of Broiler Chicken under Acute Heat Stress Condition as Revealed by Molecular Analysis of 16S rRNA Genes
Authors: Lan Pham Thi Ngoc , Sakamoto Mitsuo , Benno Yoshimi 
Published: November 2013

AbstractWe examined the effects of probiotic Lactobacillus strains of Lactobacillus agilis JCM 1048 and Lactobacillus salivarius subsp. salicinius JCM 1230 on jejunal and cecal microbiota of broiler chicken under heat stress condition using terminal restriction fragment length polymorphism (T‐RFLP) analysis. The jejunal bacterial community was limited to a few bacterial groups, mostly Lactobacillus spp. A relatively abundant and higher prevalence of Lactobacillus spp. were observed in the jejunal and cecal microbiota of the probiotic chickens compared with those of the control chickens under heat stress condition. In general, the probiotic strains did not significantly affect the abundance of L. agilis and L. salivarius in chicken intestine but clearly contributed to increasing their prevalence in the probiotic chickens. The probiotic Lactobacillus strains enriched the diversity of Lactobacillus flora in chicken jejunum and cecum by increasing the abundance and prevalence of Lactobacillus spp. inhabiting the intestine. The richness of Lactobacillus species tended to be similar among the jejunal and cecal microbiota. The bacterial community of cecum was complex and age‐dependent. The major components of the cecal microbiota were clostridia and lactobacilli. The Clostridium subcluster XIVa was the most predominant group in chicken cecum. Probiotic Lactobacillus strains restored the microbial balance and maintained the natural stability of indigenous bacterial microbiota following heat stress‐induced changes.
Comparison of Japanese Isolates of Coxiella burnetii by PCR‐RFLP and Sequence Analysis
Authors: Andoh Masako , Nagaoka Hiromi , Yamaguchi Tsuyoshi , Fukushi Hideto , Hirai Katsuya 
Published: November 2013

AbstractThe genetic variation of Japanese isolates of Coxiella burnetii, the agent of Q fever, was found for the first time. Forty‐nine out of 72 isolates had the chronic pattern of the isocitrate hydrogenase gene. Sequence analysis revealed that the isolates have a specific nucleotide sequence. The putative amino acid sequence was the same as that of chronic reference strains. These results suggest the variation of C. burnetii isolates in Japan.
B1‐B Cells Are the Main Antigen Presenting Cells in CpG‐ODN‐Stimulated Peritoneal Exudate Cells
Authors: Bamba Hiromichi , Ishigaki Hirohito , Ishida Hideaki , Kajino Kiichi , Fujiyama Yoshihide , Ogasawara Kazumasa 
Published: November 2013

AbstractPeritoneal exudate cells (PEC) have long been used as antigen presenting cells (APC), because they have been considered to contain mainly macrophages. However, it is still unclear specifically which cells of the peritoneal exudate function as APC. Herein, we focused on macrophages and B1‐B cells of the PEC and examined their APC function and cytokine production. B1‐B cells purified from PEC functioned effectively as APC after CpG‐stimulation and mainly produced IL‐10. In contrast, macrophages purified from PEC were not able to present incorporated antigens to T cells, despite the production of IL‐12 and expression of co‐stimulatory molecules after CpG stimulation. These results suggest that previously held ideas regarding the functions of the mixture of cells in the PEC need to re‐evaluated. In summary, the antigen presenting function of PEC was mainly attributed to B1‐B cells and immunoenhancing cytokine production was dominantly derived from peritoneal macrophages.