bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular Epidemiology of Salmonella enterica Serovar Enteritidis Isolated in Taiwan
Authors: Su Lin‐hui , Chiu Cheng‐hsun , Wu Tsu‐lan , Chu Chishih , Chia Ju‐hsin , Kuo An‐jing , Lee Chia‐ching , Sun Chien‐feng , Ou Jonathan T. 
Published: November 2013

AbstractIncidence of Salmonella enterica serovar Enteritidis infection seems to be on the rise in Taiwan, and therefore, the characteristics of the isolate, including genotypes, were epidemiologically investigated. Of the 71 clinical strains isolated in 1997–1999, 61 (86%) remained susceptible to the eight antibiotics tested, while the remaining ten, eight of which were isolated in 1999, were resistant to one to three of the agents including three multiply resistant strains. The majority, 69 or 97% of the isolates, harbored a 60‐kb spvC gene‐carrying virulence plasmid and 12 of them harbored one or two additional various‐sized plasmids. Strains with more than one plasmid were isolated mostly in 1999. Pulse‐field gel electrophoresis (PFGE) revealed three major genotypes (Types A, B and C), in which Type A was the predominant type. Of the 68 Type A, which contained 8 subtypes, 59 (83%) belonged to only two subtypes. Similar results were obtained with a PCR‐based typing method, the infrequent‐restriction‐site (IRS) PCR. All four methods detected types that were rarely seen before and most of these were of recent isolates, indicating that these unusual types were new or strains of foreign origin. Though all four methods discriminated types well, PFGE and IRS‐PCR showed higher sensitivity for classification. Between the two, the latter, though less discriminatory than PFGE, seems the method of choice, since it is simpler, less time‐consuming and above all easy to perform.
Distribution of Rat C5a Anaphylatoxin Receptor
Authors: Akatsu Hiroyasu , Abe Masayoshi , Miwa Takashi , Tateyama Hisashi , Maeda Seiji , Okada Noriko , Kojima Kiyohide , Okada Hidechika 
Published: November 2013

AbstractThe anaphylatoxin, complement 5a (C5a), plays a key role in mediating various inflammatory reactions following complement activation. Several investigators have reported that C5a receptor (C5aR) is expressed in non‐myeloid cells under certain conditions or in different cell lines. In our study, the abundance of C5aR‐positive myeloid cells in rats depended on the organs examined. C5aR was usually expressed at the site of exposure to pathogens, such as in salivary gland or lung, and was up‐regulated in liver in the inflammatory state induced by lipopolysaccharide (LPS) administration. Furthermore, the increased expression of C5aR antigen was not accompanied by an increase in C5aR mRNA in Kupffer cells following LPS challenge.
Identification of Genetic Determinants That Regulate Tumorigenicity of Friend Murine Leukemia Virus in Rats
Authors: Takase‐Yoden Sayaka , Watanabe Rihito 
Published: November 2013

AbstractA neuropathogenic variant of Friend murine leukemia virus (FrMLV), clone A8, has been shown to cause thymoma and infiltration of leukemic cells to organs at 7–8 weeks post‐infection in rats with a more rapid progression than clone 57. We have previously reported that the determinant for induction of aggressive leukemia in rats is located in the ClaI‐AatII fragment containing the long terminal repeat (LTR) and the 5′ half of the 5′ leader sequence of A8 virus. Further studies of chimeric viruses restricted the determinant for the induction of thymoma to only the 0.6‐kb ClaI‐KpnI fragment of A8. This fragment contains a 0.1 kb region of the 3′ terminus of the env gene, the intergenic region, the U3, and the 5′ half of the R region in the LTR. Major differences in the fragment between A8 and 57 viruses were found in the U3 region, especially in the enhancer motifs. These results indicate that the enhancer region of A8‐LTR contributes to the manifestation of thymoma with rapid progression in rats.
Effect of Environmental Conditions on Proteins Secreted by Enterohemorrhagic Escherichia coli O26:H11
Authors: Yoh Myonsun , Bi Zhenqing , Matsuyama Junko , Nagayama Kenichi , Honda Takeshi 
Published: November 2013

AbstractInfections due to Shiga toxin‐producing Escherichia coli (STEC) are responsible for severe diarrheal diseases in humans, and these bacteria have recently emerged as a leading cause of renal failure and encephalitis in children and the aged. In this study, we examined the environment‐dependent production of proteins secreted from a strain of STEC O26:H11 by trichloroacetic acid precipitation, SDS‐PAGE, Western blotting and N‐terminal amino acid sequence analysis. Growth of bacteria in essential minimum medium (M9) led to the detection of secreted proteins of 104, 80, 40, 37 and 25 kDa (P104, P80, P40, P37 and P25, respectively). When grown in serum‐free MEM, only P104, P40, P37 and P25 were observed in supernatant fluids. Growth of the bacteria in Luria‐Bertani broth (LB) enhanced the expression of P104, but the productions of the other proteins were remarkably reduced. CO2 increased the secretion of P80 and P37, but reduced the production of P104. N‐terminal amino acid sequencing revealed that P104 was EspP of STEC, which was homologous to EspC of enteropathogenic Escherichia coli (EPEC), and both proteins belong to a subclass of the IgA protease family. P80, which was identified as EspE of STEC, was homologous to Tir of EPEC. P40, P37 and P25 were found to be highly homologous to the similarly sized EspD, EspB and EspA proteins, previously detected in culture supernatants of EPEC. Those proteins are thought to be STEC virulence factors. Sera were obtained from two patients, one with colitis and another with hemolytic uremic syndrome (HUS), caused by STEC O157:H7, to study immune response to secreted proteins. Our results suggested that Tir caused immune response following STEC disease.
Transposon‐Induced Norfloxacin‐Sensitive Mutants of Bacteroides thetaiotaomicron
Authors: Ueda Ohmi , Yoshimura Fuminobu 
Published: November 2013

AbstractTo elucidate the mechanism of norfloxacin (a fluoroquinolone) resistance of Bacteroides thetaiotaomicron, a member of the B. fragilis group, we isolated transposon‐induced mutants sensitive to this agent using Tn4351. Four norfloxacin‐sensitive mutants showed reduced levels of resistance, at least, to ethidium bromide. Cloning and sequencing of three chromosomal fragments adjacent to Tn4351 from the mutants revealed that two partial open reading frames (orfs) were disrupted by a transposon. Amino acid sequences of partial orf products had strong homologies to those of Escherichia coli RecB and B. ovatus transketolase. Two mutants carried a recB homolog inserted by Tn4351 together with R751 (cointegration) and by itself (simple transposition) at the amino‐ and carboxyl‐terminal portions, respectively. Since mutations in recB produce E. coli cells sensitive to DNA‐damaging treatments by quinolones, it is concluded that decreases of the minimum inhibitory concentrations (MICs) of the agents for B. thetaiotaomicron resulted from disruption of the recB homolog. Another mutant carried a transketolase gene inserted by Tn4351. There is no reasonable explanation why disruption of the transketolase gene caused a decrease of the MIC of norfloxacin for this organism, although Streptococcus pneumoniae RecP related to DNA recombination was reported to be transketolase.
Isolation of Escherichia coli O157:H7 from Dung Beetles Catharsius molossus
Authors: Xu Jianguo , Liu Qiyong , Jing Huaiqi , Pang Bo , Yang Jinchuan , Zhao Guangfa , Li Hongwei 
Published: November 2013

AbstractIn an epidemiological survey, Escherichia coli O157:H7 was isolated from the intestine 4 of 113 dung beetle Catharsius molossus captured below ground at Tongshan County, Jiangsu Province of China. In parallel, 10 strains of E. coli O157:H7 were isolated from fecal samples of 383 diarrhea patients from the same region. Most importantly, using pulsed field gel electrophoresis (PFGE) of chromosomal DNA restriction fragments and PCR method, we found that the PFGE pattern and virulence genes of beetle isolates were identical to those of the human isolates, such as Shiga‐toxins (stx) and enterohemorrhagic Escherichia coli hemolysin A (EHEC‐hlyA). Therefore, dung beetle might acquire pathogenic E. coli O157:H7 through contact with feces of domestic animals.
Individuals Exhibiting Conspicuous Nevi (Lentigo Simplex) Are Resistant to Allergic Rhinitis/Conjunctivitis (Pollinosis), but Those Who Do Not Show Increased Susceptibility to Pollinosis
Authors: Awaya Akira , Watanabe Ken‐ichi , Kato Shinsuke 
Published: November 2013

AbstractWe investigated 115 individuals in Tokyo from two groups of persons with conspicuous nevi on the head, neck, and arms, etc. (nevus‐positive group) and persons who do not exhibit such nevi (nevi‐negative group) interviewing them to determine whether they suffered from allergic rhinitis/conjunctivitis (pollinosis) and examining them for the presence and extent of nevi. There was a distinct correlation: only 5 persons among 85 individuals of the nevus‐positive group suffered from pollinosis, and all 30 people in the nevus‐negative group had pollinosis. Thus, individuals exhibiting conspicuous nevi are resistant to pollinosis, but those who do not show increased susceptibility to pollinosis.
Screening Method for Salmonella enterica Serovar Typhi and Serovar Paratyphi A with Reduced Susceptibility to Fluoroquinolones by PCR‐Restriction Fragment Length Polymorphism
Authors: Hirose Kenji , Tamura Kazumichi , Watanabe Haruo 
Published: November 2013

AbstractSalmonella enterica serovar Typhi and serovar Paratyphi A with reduced susceptibility to fluoroquinolones (MICs of ciprofloxacin, 0.25 to 2 μg/ml) have a mutation at codon either Ser‐83 or Asp‐87 of gyrA gene. A screening method by PCR‐restriction fragment length polymorphism (PCR‐RFLP) was designed to screen the mutations at codon Ser‐83 and Asp‐87 of the gyrA gene of S. enterica serovar Typhi and serovar Paratyphi A clinical isolates. This method successfully screened the gyrA mutations of S. enterica serovar Typhi and serovar Paratyphi A with reduced susceptibility to fluoroquinolones.
Degradation Process of Mycobacterium leprae Cells in Infected Tissue Examined by the Freeze‐Substitution Method in Electron Microscopy
Authors: Amako Kazunobu , Takade Akemi , Umeda Akiko , Matsuoka Masanori , Yoshida Shin‐ichi , Nakamura Masahiro 
Published: November 2013

AbstractMycobacterium leprae cells (strain Thai‐53) harvested from infected mouse foot pads were examined by electron microscopy using the freeze‐substitution technique. The population of M. leprae cells from the infected tissue consisted of a large number of degraded cells and a few normal cells. These thin sectioned cell profiles could be categorized into four groups depending on the alteration of the membrane structures, and the degradation process is considered to occur in stages, namely from stages 1 to 3. These are the normal cells with an asymmetrical membrane, a seemingly normal cell but with a symmetrical membrane (stage 1), a cell possessing contracted and highly concentrated cytoplasm with a membrane (stage 2), and a cell that has lost its membrane (stage 3). The peptidoglycan layer was found to remain intact in these cell groups.
Disruption of the Human Pathogenic Yeast Candida albicans Catalase Gene Decreases Survival in Mouse‐Model Infection and Elevates Susceptibility to Higher Temperature and to Detergents
Authors: Nakagawa Yoshiyuki , Kanbe Toshio , Mizuguchi Ikuyo 
Published: November 2013

AbstractCatalase‐deficient strains of the human pathogenic yeast Candida albicans were constructed using the URA‐blaster method. The disruptant was viable and grew normally in an ordinary culture condition, but became extremely sensitive to treatment with hydrogen peroxide. No catalase activity was observed in a catalase (CCT)‐gene‐disrupted strain, 1F5‐4‐1, suggesting that there were no other catalase or catalase‐like enzymes in this yeast. The disruptant was shown to be sensitive to higher temperature and to low concentrations of SDS, NP‐40, or Triton X‐100. After a wild‐type CCT gene was reintroduced into the disruptant, catalase activity was restored and the strain became moderately sensitive to treatment with hydrogen peroxide. However, neither the temperature sensitivity nor the susceptibility to SDS observed in the disruptant was restored in the CCT‐reintroduced strain. A model infection experiment using wild‐type and dCCT strains showed that the disruptants disappeared more rapidly than the wild‐type strain in mouse liver, lung, and spleen. These results suggest that the catalase plays a significant role in survival in the host immune system and thus leads this organism to establish infection in the host.
Chimeric Human/Murine Monoclonal IgM Antibodies to HIV‐1 Nef Antigen Expressed on Chronically Infected Cells
Authors: Kawai Masahiro , He Lianying , Kawamura Takeshi , Omoto Shinya , Fujii Yoichi R. , Okada Noriko 
Published: November 2013

AbstractHuman IgM antibody (Ab) to gangliosides induced cytolysis of HIV‐1‐infected cells by homologous human complement. We expected that any human IgM Ab reactive with HIV‐1 infected cells could cause complement‐mediated cytolysis. The trans‐chromosome mouse (TC mouse) contains human chromosomes harboring genes responsible for immunoglobulin production. Spleen cells from TC mice immunized with recombinant Nef were fused with mouse myeloma cells to generate hybridomas, and we selected those that produced human μ‐chain‐positive Abs reactive with Nef fixed on an ELISA plate. However, the L‐chain of the monoclonal Abs (mAbs) were murine λ in type and were chimeric, and we could not succeed in obtaining mAb with human μ‐ and human κ‐chains. The chimeric mAbs reacted with the HIV‐1 infected cells as seen with flow cytometric analysis, and the surface expression of Nef was also detectable on chronically infected OM10.1 cells which had no detectable gp120. However, although the reaction of the chimeric IgM mAb with HIV‐1‐infected MOLT4 cells induced C3 deposition on cell surfaces on incubation with fresh human serum, the cells remained unlysed, as determined by 51Cr release assay. The amount of Nef antigen on the cells might not have been high enough to overcome the function of HRF20 (CD59) that restricts formation of membrane attack complexes of homologous complement. However, combination of anti‐Nef IgM mAb with other IgM mAbs reactive with the surface of HIV‐1‐infected cells may induce a synergistic effect in complement mediated cytolysis.
The Urease Gene Cluster of Vibrio parahaemolyticus Does Not Influence the Expression of the Thermostable Direct Hemolysin (TDH) Gene or the TDH‐Related Hemolysin Gene
Authors: Nakaguchi Yoshitsugu , Okuda Jun , Iida Tetsuya , Nishibuchi Mitsuaki 
Published: November 2013

AbstractIn order to investigate why the thermostable direct hemolysin (TDH) and the TDH‐related hemolysin (TRH) of Vibrio parahaemolyticus are produced at low levels from urease‐positive strains, the effect of the functional urease gene cluster of V. parahaemolyticus on the expression of the tdh and trh genes was examined. Transcriptional lacZ fusions with the tdh1, tdh2, trh1 and trh2 genes representing variants of the tdh and trh genes were integrated into the chromosome of an Escherichia coli strain and a urease‐negative V. parahaemolyticus strain. The plasmid‐borne urease gene cluster introduced and expressed in these constructs did not affect expression of any of the fusion genes. The amount of TDH produced from a Kanagawa phenomenon‐positive V. parahaemolyticus did not change by introduction of the urease gene cluster either. It was concluded therefore that the urease gene cluster is not involved in the regulation of tdh and trh expression.
An Effective Sporicidal Reagent against Bacillus subtilis Spores
Authors: Kida Nori , Mochizuki Yasushi , Taguchi Fumiaki 
Published: November 2013

AbstractWe developed a reagent which showed significant sporicidal activity against Bacillus subtilis spores. This reagent was composed of ethylenediaminetetraacetic acid, disodium salt (EDTA‐2Na), ferric chloride hexahydrate (FeCl3 · 6H2O) and ethanol (tentatively designated as the ethanol reagent). The ethanol reagent showed pH‐ and temperature‐dependent sporicidal activity. At pH 0.3, its activity was almost the same as that of 0.05% sodium hypochlorite at 20 C and was higher at 37 C than at 20 C. The activity of the ethanol reagent was similar both with and without 10% serum. The ethanol reagent might be applicable for disinfecting Bacillus spores.
Preferential Detection of Pro‐Carboxypeptidase R by Enzyme‐Linked Immunosorbent Assay
Authors: Tani Sayuri , Akatsu Hiroyasu , Ishikawa Yasushige , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractWe generated two monoclonal antibodies (mAbs), 2A16 and 10G1, against pro‐carboxypeptidase R (proCPR), also known as thrombin activatable fibrinolysis inhibitor (TAFI). By use of these mAbs, we developed a sandwich enzyme‐linked immunosorbent assay (ELISA) system to detect proCPR. Since the amount of the antigen detectable by the ELISA was essentially the same in fresh plasma and serum incubated at 37 C for 1 hr, we concluded that the ELISA system detected not only proCPR, but also inactivated CPR generated from proCPR. However, an appreciable amount of proCPR remained unactivated in serum. For extensive activation of proCPR in plasma, thrombin and thrombomodulin complexes (T‐TM) can be used together with CaCl2. Following extensive conversion of proCPR to CPR by T‐TM and CaCl2, converting plasma to serum (T‐TM serum), antigenicity became undetectable by ELISA. Further analysis revealed that 2A16 reacts only with proCPR although 10G1 reacts with proCPR, active CPR and inactivated CPR. Therefore, we concluded that the ELISA system preferentially detects proCPR and not CPR. Our sandwich ELISA system utilizing 2A16 and 10G1 provides a suitable method for detecting proCPR and can be used to determine levels of proCPR in plasma samples from patients.
Morphology of the Type Strain of Bacillus anthracis EY 3169T=ATCC 14578T Grown Either Aerobically or Anaerobically on Agar Plates —Observation by Light and Laser Microscopes—
Authors: Yabuuchi Eiko , Koseki Masayuki 
Published: November 2013

AbstractGrowth characteristics including cell‐arrangement of the type strain of Bacillus anthracis EY 3169T=ATCC 14578T grown on agar plates in level 3 laboratory were observed by both light and laser microscopes. Small daughter colonies appeared on parent colonies grown on 5% sheep blood or chocolate agar plates after 12 days incubation at room temperature. Daughter colonies, stained by Wirtz‐Conklin method, were composed with vegetative cells and spores. Growth of daughter colonies might be supported by the debris of cells in the parent colony. Colonies grown under anaerobic conditions were flat with smooth edges, and the cells neither formed chains of any length, nor produced any spores after 25 days incubation at room temperature. It was thought that spores of B. anthracis were produced at the terminal stage of individual cell life instead of under unfavorable conditions for the organism. Air is needed for spore formation and cell‐chain formation. More nutrients, probably amino acids, are needed for anaerobic growth rather than aerobic.
Hepatitis C Virus Nonstructural Protein NS3 Binds to Sm‐D1, a Small Nuclear Ribonucleoprotein Associated with Autoimmune Disease
Authors: Iwai Atsushi , Hasumura Yasushi , Nojima Takayuki , Takegami Tsutomu 
Published: November 2013

AbstractHepatitis C virus (HCV) causes a persistent infection, chronic hepatitis, and leads to hepatocellular carcinoma. Nonstructural protein 3 (NS3) of HCV possesses protease, nucleoside triphosphatase, and helicase activities. Using the yeast two hybrid assay, we identified Sm‐D1, a host protein that binds to NS3. Sm‐D1 is a component of small nuclear ribonucleoprotein (snRNP) complexes which are associated with autoimmune disease. Sm‐D1 has Gly‐Arg (GR) repeats at the C terminus, which contains dimethylarginine modified by post‐translational modification and may constitute an immunoreactive determinant. Deletion mutants revealed that the C‐terminal region of Sm‐D1 containing the GR repeats was the binding region for NS3, and the expression feature of Sm‐D1 was affected by co‐expression of NS3. Immunostaining assay demonstrated that NS3 was also present in the nucleus of cells overexpressing Sm‐D1, although it was usually found in cytoplasm. The localization of NS3 could change following interaction with Sm‐D1 and affect the function of Sm‐D1.
Improvements of PCR‐Based Identification Targeting the DNA Topoisomerase II Gene to Determine Major Species of the Opportunistic Fungi Candida and Aspergillus fumigatus
Authors: Kanbe Toshio , Arishima Takuro , Horii Toshinobu , Kikuchi Akihiko 
Published: November 2013

AbstractFor the simple and rapid detection/identification of major pathogenic fungal species such as Candida albicans, C. tropicalis, C. parapsilosis, C. glabrata and Aspergillus fumigatus, common primers for these species and specific primers for each species, designed on the basis on the genomic nucleotide sequences of the DNA topoisomerase II genes, were prepared and tested for their specificities in PCR amplifications. Twelve specific primers were pooled and designated PsVI. Genomic DNAs were amplified by the common primer pair, and followed by PCR amplification using PsVI. Using PsVI, six unique DNA fragments, all of which corresponded to a Candida or A. fumigatus species, were specifically and acceptably amplified from each template DNA even in the presence of other DNAs. Similarly, the results of identification of clinical samples based on the PCR amplification coincided with those of conventional identification techniques. The sensitivities of the direct PCR and the nested PCR using PsVI were found to be 1,000 and 50 yeast cells, respectively.
Herpes Simplex Virus UL14 Protein Blocks Apoptosis
Authors: Yamauchi Yohei , Daikoku Tohru , Goshima Fumi , Nishiyama Yukihiro 
Published: November 2013

AbstractWe report that an HSV‐2 UL14 protein expressing cell line (14/HEp‐2) was more resistant to apoptosis induced by osmotic shock and certain drugs than its parental cell line. Furthermore, HSV‐1 UL14 protein deletion virus (UL14D) showed weaker inhibition of apoptosis compared to the rescued virus UL14R. The protein' s anti‐apoptotic function may derive from its heat shock protein‐like properties.
Depolymerization of Actin Filament by Cytochalasin E Induces Interleukin‐8 Production and Up‐Regulates CD54 in the HeLa Epithelial Cell Line
Authors: Ikewaki Nobunao , Yamada Akira , Inoko Hidetoshi 
Published: November 2013

AbstractWe previously reported that the depolymerization of actin filament by cytochalasin E enhances low affinity Fcε receptor II (CD23) expression on the human monocyte‐like cell line, U937 (J. Clin. Immunol. 20: 235, 2000). In this study, we found that cytochalasin E strongly induces interleukin‐8 through an epithelial cell line, HeLa, in dose‐ and time‐dependent manners as assessed by enzyme‐linked immunoassay and reverse transcription‐polymerase chain reaction techniques. In addition, interleukin‐8 production in the HeLa cells cultured with cytochalasin E was blocked in the presence of protein kinase C inhibitors, Go6976 and H‐7. On the other hand, it was found that CD54 (intercellular adhesion molecule‐1; ICAM‐1) expression on the HeLa cells and the secretion of soluble CD54 were significantly up‐regulated after culturing with cytochalasin E, and that these up‐regulations of CD54 were also suppressed by Go6976. Taken together, these findings indicate that cytochalasin E activates protein kinase C under the depolymerization of actin filament, leading to the induction of interleukin‐8 production and the up‐regulation of CD54 in HeLa cells.