bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interferon Gamma Response to Combinations 38 kDa/CFP‐10, 38 kDa/MPT‐64, ESAT‐6/MPT‐64 and ESAT‐6/CFP‐10, Each Related to a Single Recombinant Protein of Mycobacterium tuberculosis in Individuals from Tuberculosis Endemic Areas
Authors: Tavares Ricardo C. Oliveira , Salgado Jorge , Moreira Valéria Barbosa , Ferreira Mônica Antonia S. , Mello Fernanda C. Queiroz , Leung Janaína W. , Fonseca Leila de Souza , Spallek Ralf , Singh Mahaviir , Saad Maria Helena F. 
Published: November 2013

AbstractSeveral antigens of Mycobacterium tuberculosis have been identified and specificity to one or multiple antigens could determine the distinction between protective and pathogenic host reaction. Therefore T cell immune response to combinations 38 kDa/CFP‐10, 38 kDa/MPT‐64, ESAT‐6/MPT‐64 and ESAT‐6/CFP‐10 (each related to a single protein of Mycobacterium tuberculosis) in individuals from tuberculosis endemic areas have been examined. ELISA was used to detect IFN‐γ production in PBMC priming with single proteins and combinations in a panel of 105 individuals: 38 tuberculosis patients (6 untreated and 32 treated) and 67 healthy controls with tuberculin skin test positive or negative (TST). Brazilian TB patients highly recognized ESAT‐6 (66%), but combinations improved response in the following order: ESAT‐6/MPT‐64 (89%) > ESAT‐6/CFP‐10 (73%) > 38 kDa/CFP‐10 (70%), the last combination showing the highest specificity (TST+=42% and TST–=83%). Average IFN‐γ production in TB patients was significantly higher for 38 kDa/CFP‐10 (P=0.012) and 38 kDa/MPT‐64 (P<0.035), when compared to single antigens. None of the combinations was able to discriminate TB patients from TST+ controls; however, 38 kDa/CFP‐10 displayed a borderline significance (P=0.053). Similar to the ESAT‐6/CFP‐10 combination, IFN‐γ response to 38 kDa/CFP‐10 showed an increased tendency in treated patients, although not significant (P=0.16). We demonstrated for the first time that 38 kDa/CFP‐10 had prediction sensitivity for TB patients similar to the ESAT‐6/CFP‐10 combination and also significant response improvement related to the single proteins with more selective reactivity among TST‐positive individuals, which could be of potential interest for diagnostic evaluation for tuberculosis infection.
Increased Inhibitory Capacity of an Anti‐C5a Complementary Peptide Following Acetylation of N‐terminal Alanine
Authors: Okada Noriko , Asai Suzuka , Hotta Aya , Miura Noriko , Ohno Naohito , Farkas Imre , Hau Lewis , Okada Hidechika 
Published: November 2013

AbstractAmino acids 37 to 53 (RAARISLGPRCIKAFTE) of C5a anaphylatoxin form an essential region for C5a function. To target this sequence, we generated a complementary peptide (ASGAPAPGPAGPLRPMF) designated PepA which has a potent inhibitory effect on C5a activity. By introducing an acetyl group at the N‐terminal alanine of PepA, an acetylated form was generated which was designated AcPepA. The acetylation resulted in increased inhibition of C5a stimulation of neutrophils as determined by Ca influx. Furthermore, AcPepA partially inhibited the lethal shock induced in mice by intravenous administration of Candida albicans water‐soluble mannoprotein‐β‐glucan complex. In addition, local skin inflammation in rats caused by an anti‐Crry monoclonal antibody was suppressed when AcPepA and the antibody were injected together, while PepA had little inhibitory capacity. The potent inhibitory capacity of AcPepA was also confirmed by a skin reaction of guinea pigs inoculated with recombinant human C5a together with AcPepA.
C Terminal Half Fragment (50 kDa) of Heavy Chain Components of Clostridium botulinum Type C and D Neurotoxins Can Be Used as an Effective Vaccine
Authors: Lee Jae‐Chul , Hwang Hyun‐Jung , Sakaguchi Yoshihiko , Yamamoto Yumiko , Arimitsu Hideyuki , Tsuji Takao , Watanabe Toshihiro , Ohyama Tohru , Tsuchiya Tomofusa , Oguma Keiji 
Published: November 2013

AbstractRecombinant whole heavy chains (H, 100 kDa) and their N‐terminal (Hn, 50 kDa) and C‐terminal (Hc, 50 kDa) half fragments of Clostridium botulinum type C and D neurotoxins were expressed as glutathione S‐transferase (GST) fusion proteins in Escherichia coli. GST eliminated‐preparations of H (10 μg), Hn (5 μg), Hc (5 μg), or a mixture of Hn (5 μg) and Hc (5 μg) of types C and D were mixed with an equal volume of adjuvant, and then were twice injected into mice subcutaneously. After immunization, the mice were challenged with up to 106 the minimum lethal doses (MLD)/0.5 ml of C or D toxin, the type of which was same as that of the immunogens. All of the mice immunized with antigens except for Hn survived against 105 to 106 MLD/0.5 ml of the toxins, but the mice immunized with Hn were killed by 100 MLD/0.5 ml. The mice immunized with a mixture of C‐Hc and D‐Hc, each 5 μg, also showed a high level of resistance against both C and D toxins. Antibody levels immunized with GST fused‐ or GST eliminated‐preparation were quite similar. These results indicate that recombinant GST‐fused Hc can be used as a safe and effective vaccine for type C and D botulism in animals. It also became clear that one time inoculation with a large amount of C‐Hc or D‐Hc, 100 μg, is useful for vaccine trials in mice.
Serum Withdrawal‐Induced Apoptosis in ZrchI Prion Protein (PrP) Gene‐Deficient Neuronal Cell Line Is Suppressed by PrP, Independent of Doppel
Authors: Nishimura Takuya , Sakudo Akikazu , Hashiyama Yoriko , Yachi Akiko , Saeki Keiichi , Matsumoto Yoshitsugu , Ogawa Masaharu , Sakaguchi Suehiro , Itohara Shigeyoshi , Onodera Takashi 
Published: November 2013

AbstractPrevious studies have shown that cellular prion protein (PrPC) plays anti‐apoptotic and anti‐oxidative role against cell death induced by serum‐deprivation (SDP) in an immortalized prion protein gene‐deficient neuronal cell line derived from Rikn prion protein (PrP) gene‐deficient (Prnp–/–) mice, which ectopically produce excess Doppel (Dpl) (PrP‐like glycoprotein). To investigate whether PrPC inhibits apoptotic neuronal cell death without Dpl, an immortalized cell line was established from the brain of ZrchI Prnp–/– mice, which do not show ectopic expression of Dpl. The results using a ZrchI neuronal Prnp–/– cell line (NpL2) showed that PrPC potently inhibited SDP‐induced apoptotic cell death. Furthermore, PrPC expression enhanced the superoxide dismutase (SOD) activity in NpL2 cells. These results indicate that Dpl production did not affect anti‐apoptotic and anti‐oxidative functions of PrP, suggesting that PrPC may be directly correlated with protection against oxidative stress.
Biofilm Acts as a Microenvironment for Plankton‐Associated Vibrio cholerae in the Aquatic Environment of Bangladesh
Authors: Islam Mohammad Sirajul , Jahid Mohammad Iqbal Kabir , Rahman Mohammad Majibur , Rahman Mohammed Ziaur , Islam Mohammad Shafiqul , Kabir Mohammad Shahidul , Sack David Allen , Schoolnik Gary K. 
Published: November 2013

AbstractThe role of biofilm as a microenvironment of plankton‐associated Vibrio cholerae was investigated using plexiglass as a bait A total of 72 biofilm samples were tested using culture, direct fluorescent antibody (DFA) and molecular techniques following standard procedures. Culturable V. cholerae (smooth and rugose variants) were isolated from 33% of the samples. V. cholerae O1 were detected by FA technique throughout the year except April and June. All V. cholerae O1 isolates were positive for tcpA, ctxA and ace genes while V. cholerae non‐O1, non‐O139 isolates lacked these genes. V. cholerae O1 (both Inaba and Ogawa) strains had identical ribotype pattern (R1), but V. cholerae non‐O1, non‐O139 had different ribotype patterns. All V. cholerae O1 strains were resistant to vibrio‐static compound (O/129). All V. cholerae O1 except one were resistant to trimethoprime‐sulphamethoxazole, streptomycin, nalidixic acid and furazolidone but sensitive to ciprofloxacin, and tetracycline. This study indicates that plexiglass can act as a bait to form biofilm, a microenvironment that provides shelter for plankton containing V. cholerae in the aquatic environment of Bangladesh.
Distinct Function of Pseudomonas aeruginosa Type IV Pili Disclosed in the Bacterial Pass‐Through of Membrane Filter with Smaller Pore Sizes
Authors: Hasegawa Hiroyuki , Tanikawa Taichiro , Nozawa Takashi , Nakazawa Kentaro , Nakagawa Yoji , Matsuyama Tohey 
Published: November 2013

AbstractMembrane filter pass‐through ability of Pseudomonas aeruginosa was analyzed with isogenic mutants. A flagellum‐deficient fliC mutant required two‐times longer time (12 hr) to pass through a 0.45‐μm pore size filter. With 0.3‐ and 0.22‐μm filters, however, the fliC mutant showed no remarkable disability. Meanwhile a pilA mutant defective in twitching motility failed to pass through the 0.22‐μm filter. Complementation of the mutant with pilA gene on a plasmid restored the twitching motility and the 0.22‐μm filter pass‐through activity. Thus, the distinctive role of P. aeruginosa type IV pili in infiltration into finer reticulate structures was indicated.
A Novel In Vitro Pharmacokinetic/Pharmacodynamic Model Based on Two‐Compartment Open Model Used to Simulate Serum Drug Concentration‐Time Profiles
Authors: Tomita Tsutomu , Ohara‐Nemoto Yuko , Moriyama Hitomi , Ozawa Ayako , Takeda Yoshifumi , Kikuchi Ken 
Published: November 2013

AbstractAn in vitro pharmacokinetic/pharmacodynamic perfusion model that simulates a two‐compartment open model of serum drug concentration‐time profiles following intravenous bolus injection and infusion was developed and mathematically described. In the present apparatus model, flow was kept in a one‐way mode to avoid liquid traffic, and the washout effect seen in dilution models was overcome by embedding the tested bacteria in low melting point agarose gel. The validity of the equations and the reproducibility of the apparatus model were ascertained by simulating the concentration‐time profiles of cefazolin and fosfomycin by substitution of their pharmacokinetic parameters obtained from humans for the equations. An empirical regimen 1X(q24h) of 1 g with cefazolin administered by intravenous infusion effectively killed a Staphylococcus aureus strain. The same regimen with fosfomycin produced a marked kill‐curve with a fosfomycin‐susceptible enterohaemorrhagic Escherichia coli O157:H7, whereas considerable regrowth was observed with a resistant strain. These results indicated that the present model was able to provide a convenient and reliable method for evaluating the efficacy of antimicrobial agents administered by intravenous infusion.
Extensive Accumulation of Influenza Virus NS1 Protein in the Nuclei Causes Effective Viral Growth in Vero Cells
Authors: Ozaki Hiroichi , Kida Hiroshi 
Published: November 2013

AbstractWe previously showed that modified A/Puerto Rico/8/34 (PR8) influenza master strain had improved viral rescue and growth properties in African green monkey kidney (Vero) cell line by introducing NS gene of Vero‐adapted A/England/1/53 (vaEng53). In the present study, it was found that the NS1 protein derived from vaEng53 was extensively accumulated in the nuclei than that of PR8. This accumulation was caused by 7 amino acid differences in C‐terminal region of NS1 protein. These results suggest that specific accumulation of NS1 protein may contribute to efficient viral replication in Vero cells.
Chlamydia pneumoniae Respiratory Infections in Taiwan
Authors: Hsu Min‐Chih , Lin Tsun‐Mei , Huang Chung‐Ter , Liao Mei‐Hui , Li Shu‐Ying 
Published: November 2013

AbstractTo understand the epidemiology of Chlamydia pneumoniae acute infections in Taiwan, we collected 116 paired and 244 single sera from patients suspected of C. pneumoniae infection and conducted microimmunofluorescence test. Eighty‐three patients (83/360, 23%) met the diagnostic criteria of current C. pneumoniae infection. The C. pneumoniae infections were significantly higher in men than in women (P≤0.0001) and were most frequent in the group of 40–49 year‐olds, and the people older than 70 years old. C. pneumoniae infection often occurred in the late autumn lasting to the cold winter and in the transition period between the spring and summer.
Neutralizing Activity of Polyvalent Gb3, Gb2 and Galacto‐Trehalose Models against Shiga Toxins
Authors: Neri Paola , Nagano Saori Itoh , Yokoyama Shin‐ichiro , Dohi Hirofumi , Kobayashi Kazukiyo , Miura Tsuyoshi , Inazu Toshiyuki , Sugiyama Tsuyoshi , Nishida Yoshihiro , Mori Hiroshi 
Published: November 2013

AbstractShiga toxin (Stx) is one of the most critical factors in the development of hemolytic uremic syndrome and other systemic complications following enterohemorrhagic Escherichia coli (EHEC) infection. Substances neutralizing Stx by interfering with toxin‐receptor binding have been explored as therapeutic candidates for EHEC infection. In this study, we examined globotriaosyl (Gb3), galabiosyl (Gb2) and galacto‐trehalose, each of which was synthetically conjugated with a polyacrylamide backbone, for Stx‐neutralizing activity. Galacto‐trehalose was designed as a Gb2 mimicking, unnatural Stx‐ligand that was expected to show tolerance to enzymatic degradation in vivo. Galacto‐trehalose copolymer showed neutralizing activity against Stx‐1 but not Stx‐2 in a HeLa cell cytotoxicity assay. It was thought that galacto‐trehalose copolymer could be a lead compound for the treatment of Stx‐mediated diseases, although it requires modification to show neutralizing activity to Stx‐2. The Gb3 copolymer with high sugar unit density showed stronger neutralizing activity against Stx‐2 than those with lower density. However, the density‐dependency of the neutralizing activity was less obvious against Stx‐1. Intravenous administration of the Gb3 copolymer prevented death in mice lethally infected with Stx‐1‐ and Stx‐2‐producing E. coli O157:H7. Thus, we demonstrated that the artificial Gb3 copolymer could neutralize Stx‐1 and the more clinically relevant Stx‐2 in vitro and effectively inhibit Stx toxicity in vivo.
Failure of Multinucleated Giant Cell Formation in K562 Cells Infected with Newcastle Disease Virus and Human Parainfluenza Type 2 Virus
Authors: Yamakawa Izumi , Tsurudome Masato , Kawano Mitsuo , Nishio Machiko , Komada Hiroshi , Ito Morihiro , Uji Yukitaka , Ito Yasuhiko 
Published: November 2013

AbstractWhen K562 cells were infected with Newcastle disease virus (NDV) or human parainfluenza type 2 virus (hPIV‐2), polykaryocyte formation could not be detected. Failure of multinucleated giant cell formation in K562 cells infected with either NDV or hPIV‐2 is due to disturbance of the viral envelope‐cell fusion step or to defect in the cell‐cell fusion step, respectively. Especially, NDV completely replicated in K562 cells, and the hemagglutinin‐neuraminidase and fusion proteins expressed on the cell surface of NDV‐infected K562 cell were fully functional for fusion inducing activity. Therefore, the cell membranes of K562 cells are considered to be resistant to virus‐induced cell fusion. Membrane fusion is regulated by many host factors including membrane fluidity, cytoskeletal systems, and fusion regulatory proteins system. An unknown regulatory mechanism of virus‐induced cell fusion may function on the cell surface of K562 cells.
Genetic Characterization of Thermal Tolerance in Enterobacter sakazakii
Authors: Asakura Hiroshi , Morita‐Ishihara Tomoko , Yamamoto Shigeki , Igimi Shizunobu 
Published: November 2013

AbstractEnterobacter sakazakii is an opportunistic pathogen that causes meningitis and necrotizing enterocolitis in neonates. Here we characterized the thermal tolerance of E. sakazakii isolates obtained from powdered infant formula and other food products in Japan. Isolates were categorized into three classes according to their thermal tolerance, and differential gene expression analysis showed that the heat‐resistant clones expressed a higher level of infB (which encodes a translation initiation factor), than did the heat‐sensitive isolates. Gene expression and DNA polymorphism analyses suggested that this gene target might be useful to unequivocally detect and identify heat‐resistant clones, permitting epidemiological surveillance for this pathogen.
Rhamnolipid‐Dependent Spreading Growth of Pseudomonas aeruginosa on a High‐Agar Medium: Marked Enhancement under CO2‐Rich Anaerobic Conditions
Authors: Nozawa Takashi , Tanikawa Taichiro , Hasegawa Hiroyuki , Takahashi Chihiro , Ando Yumi , Matsushita Mitsugu , Nakagawa Yoji , Matsuyama Tohey 
Published: November 2013

AbstractAnaerobiosis of Pseudomonas aeruginosa in infected organs is now gaining attention as a unique physiological feature. After anaerobic cultivation of P. aeruginosa wild type strain PAO1 T, we noticed an unexpectedly expanding colony on a 1.5% agar medium. The basic factors involved in this spreading growth were investigated by growing the PAO1 T strain and its isogenic mutants on a Davis high‐agar minimal synthetic medium under various experimental conditions. The most promotive environment for this spreading growth was an O2‐depleted 8% CO2 condition. From mutational analysis of this spreading growth, flagella and type IV pili were shown to be ancillary factors for this bacterial activity. On the other hand, a rhamnolipid‐deficient rhlA mutant TR failed to exhibit spreading growth on a high‐agar medium. Complementation of the gene defect of the mutant TR with a plasmid carrying the rhlAB operon resulted in the restoration of the spreading growth. In addition, an external supply of rhamnolipid or other surfactants (surfactin from Bacillus subtilis or artificial product Tween 80) also restored the spreading growth of the mutant TR. Such activity of surfactants on bacterial spreading on a hard‐agar medium was unique to P. aeruginosa under CO2‐rich anaerobic conditions.
Evaluation of Repetitive Extragenic Palindromic‐PCR for Discrimination of Fecal Escherichia coli from Humans, and Different Domestic‐ and Wild‐Animals
Authors: Mohapatra Bidyut R. , Broersma Klaas , Nordin Rick , Mazumder Asit 
Published: November 2013

AbstractThe objective of this study was to investigate the potential of repetitive extragenic palindromic anchored polymerase chain reaction (rep‐PCR) in differentiating fecal Escherichia coli isolates of human, domestic‐ and wild‐animal origin that might be used as a molecular tool to identify the possible source(s) of fecal pollution of source water. A total of 625 fecal E. coli isolates of human, 3 domestic‐ (cow, dog and horse) and 7 wild‐animal (black bear, coyote, elk, marmot, mule deer, raccoon and wolf) species were characterized by rep‐PCR DNA fingerprinting technique coupled with BOX A1R primer and discriminant analysis. Discriminant analysis of rep‐PCR DNA fingerprints of fecal E. coli isolates from 11 host sources revealed an average rate of correct classification of 79.89%, and 84.6%, 83.8%, 83.3%, 82.5%, 81.6%, 80.8%, 79.8%, 79.3%, 77.4%, 73.2% and 63.6% of elk, human, marmot, mule deer, cow, coyote, raccoon, horse, dog, wolf and black bear fecal E. coli isolates were assigned to the correct host source. These results suggest that rep‐PCR DNA fingerprinting procedures can be used as a source tracking tool for detection of human‐ as well as animal‐derived fecal contamination of water.
Methicillin Resistance in Staphylococcus aureus and Coagulase‐Negative Staphylococci: Epidemiological and Molecular Aspects
Authors: Martins André , de Lourdes R.S. Cunha Maria 
Published: November 2013

AbstractInfections caused by the genus Staphylococcus are of great importance for human health. Staphylococcus species are divided into coagulase‐positive staphylococci, represented by S. aureus, a pathogen that can cause infections of the skin and other organs in immunocompetent patients, and coagulase‐negative staphylococci (CNS) which comprise different species normally involved in infectious processes in immunocompromised patients or patients using catheters. Oxacillin has been one of the main drugs used for the treatment of staphylococcal infections; however, a large number of S. aureus and CNS isolates of nosocomial origin are resistant to this drug. Methicillin resistance is encoded by the mecA gene which is inserted in the SCCmec cassette. This cassette is a mobile genetic element consisting of five different types and several subtypes. Oxacillin‐resistant strains are detected by phenotypic and genotypic methods. Epidemiologically, methicillin‐resistant S. aureus strains can be divided into five large pandemic clones, called Brazilian, Hungarian, Iberian, New York/Japan and Pediatric. The objective of the present review was to discuss aspects of resistance, epidemiology, genetics and detection of oxacillin resistance in Staphylococcus spp., since these microorganisms are increasingly more frequent in Brazil.
Proteomic Characterization of Enterohemorrhagic Escherichia coli O157:H7 in the Oxidation‐Induced Viable but Non‐Culturable State
Authors: Asakura Hiroshi , Panutdaporn Nantika , Kawamoto Keiko , Igimi Shizunobu , Yamamoto Shigeki , Makino Sou‐ichi 
Published: November 2013

AbstractEnterohemorrhagic Escherichia coli (EHEC) O157 strain F2, a food isolate of an outbreak, is resistant to oxidative stress, but has increased stress‐sensitivity after passage through mice. The stress‐sensitive variant of F2 (designated MP37) has decreased culturability, but retains membrane integrity under stress conditions, indicating that the cells enter a viable but non‐culturable (VBNC) state. Proteomic analyses revealed that MP37 in the VBNC state had decreased levels of some oxidation‐responsive factors (AhpCF, AceF), but it markedly increased levels of outer membrane protein W (OmpW). Because F2 expressed higher levels of some ribosome‐associated proteins (RaiA, S6, Bcp) than MP37, the effect of animal passage on the induction of the VBNC state in the EHEC O157 cells might be due to ribosomal activity.
Development of an Immunochromatographic Kit for Rapid Diagnosis of H5 Avian Influenza Virus Infection
Authors: Tsuda Yoshimi , Sakoda Yoshihiro , Sakabe Saori , Mochizuki Tsuyoshi , Namba Yasuharu , Kida Hiroshi 
Published: November 2013

AbstractHighly pathogenic avian influenza (HPAI) caused by the H5N1 subtype has given rise to serious damage in poultry industries in Asia. The virus has expanded its geographical range to Europe and Africa, posing a great risk to human health as well. For the control of avian influenza, a rapid diagnosis by detecting the causative virus and identifying its subtype is essential. In the present study, a rapid diagnosis kit combining immunochromatography with enzyme immunoassay which detects the H5 HA antigen of influenza A virus was developed using newly established anti‐H5 HA monoclonal antibodies. The present kit specifically detected all of the H5 influenza viruses tested, and did not react with the other HA subtypes. H5 HA antigens were detected from swabs and tissue homogenates of chickens infected with HPAI virus strain A/chicken/Yamaguchi/7/04 (H5N1) from 2 days post inoculation. The kit showed enough sensitivity and specificity for the rapid diagnosis of HPAI.
Detection and Phylogenetic Analysis of Norovirus inCorbicula flumineain a Freshwater River in Japan
Authors: Saitoh Mika , Kimura Hirokazu , Kozawa Kunihisa , Nishio Osamu , Shoji Akira 
Published: November 2013

AbstractTo study the molecular epidemiology of noroviruses (NoVs) in bivalves residing in freshwater rivers, we detected, quantified and phylogenetically analyzed the NoV genome in purified concentrates obtained from the gills and digestive diverticula ofCorbicula flumineain a freshwater river in Gunma Prefecture, Japan. We detected the NoV genome in 35 of the 58C. flumineasamples. Based on our phylogenetic analysis, the NoV genome detected in the samples was classified into 4 genotypes (GI/1, GI/2, GI/3 and GI/4) in genogroup I and 5 genotypes (GII/3, GII/4, GII/5, GII/8 and GII/12) in genogroup II. The phylogenetic tree showed wide genetic diversity among the genogroups. In addition, more than 104copies of the NoV genome were detected in 2 of 35 samples. These results suggest that the freshwater bivalveC. flumineais a reservoir for NoVs, similar to seawater bivalves such as oysters.