bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of the ESPLINE® INFLUENZA A&B‐N Kit for the Diagnosis of Avian and Swine Influenza
Authors: Bai Gui‐Rong , Sakoda Yoshihiro , Mweene Aaron Simanyengwe , Kishida Noriko , Yamada Takashi , Minakawa Hidetaka , Kida Hiroshi 
Published: November 2013

AbstractThe ESPLINE® INFLUENZA A&B‐N kit was evaluated for its applicability to the rapid diagnosis of influenza in chickens and pigs. The kit specifically detected viral antigens in tracheal swabs and tissue homogenates of the trachea, liver, spleen, and colon of chickens inoculated with a highly pathogenic avian influenza virus strain, A/chicken/Yamaguchi/7/04 (H5N1), at 48 hr post‐inoculation (p.i.) as well as in the tracheal and cloacal swabs and tissue homogenates of dead chickens. For those infected with a low pathogenic strain, A/chicken/aq‐Y‐55/01 (H9N2), antigens were detected only in the samples from tracheal swabs and organs 1–4 days p.i. The kit also detected viral antigens in the nasal swabs of miniature pigs infected with swine and avian influenza viruses. The kit was found to be sensitive and specific enough for the rapid diagnosis of infections of influenza A virus in chickens and pigs.
Epidemiology of Typhoid Carriers among Blood Donors and Patients with Biliary, Gastrointestinal and Other Related Diseases
Authors: Vaishnavi Chetana , Kochhar Rakesh , Singh Gurpreet , Kumar Sudarshan , Singh Satnam , Singh Kartar 
Published: November 2013

AbstractEnteric fever due to Salmonella Typhi is a major public health problem. Typhoid carriers have high titres of Vi agglutinins in their sera. We worked out the baseline data for Vi agglutinins from 705 healthy blood donors (controls) by ELISA and compared it with 446 patients with biliary, gastrointestinal and other related diseases (cases). The samples were divided into five groups based on the disease condition of the patients from whom they were collected. Group A (n = 196) consisted of patients with stones in the gall bladder/common bile duct and Group B (n=27) with gall bladder carcinoma. Group C (n=33) comprised patients with carcinoma of the pancreas/ampulla, obstructive jaundice and/or cholangiocarcinoma. Group D (n = 112) had patients with acute/chronic pancreatitis, abdominal pain, intestinal obstruction, peritonitis, carcinoma oesophagus, chronic diarrhoea, gastrointestinal bleeding and dyspepsia. Group E (n=78) included patients with miscellaneous diseases. The mean absorbance value obtained for healthy subjects +3 standard deviations was taken as the cut‐off value for a positive typhoid carrier. In Group A, 10.2% samples were positive; in Group B, 7.4%; in Group C, 12.0%; in Group D, 9.8% and in Group E, 9.0%. There was a highly significant (P<0.001) increase in the presence of Vi agglutinins in the cases compared to the controls. High prevalence of typhoid carriers occurs in patients with biliary, gastrointestinal and other related diseases. Vi serology employing highly purified Vi antigen offers a practical and cost‐effective way of screening for S. Typhi carriers.
Natural Recombinant between Equine Herpesviruses 1 and 4 in the ICP4 Gene
Authors: Pagamjav Ochir , Sakata Tohru , Matsumura Tomio , Yamaguchi Tsuyoshi , Fukushi Hideto 
Published: November 2013

AbstractEquine herpesvirus 1 (EHV‐1) is a pathogen causing rhinopneumonia in young horses, abortion in mares, and myeloencephalitis in adult horses. Two types, EHV‐1 P and EHV‐1 B, have recently been dominant among 16 electropherotypes. EHV‐1 P and EHV‐1 B viruses were compared by long and accurate polymerase chain reaction (LA‐PCR) and restriction fragment length polymorphism (RFLP) analysis. Differences in restriction sites were found to be focused in ORF64, which encodes the infected cell protein 4 (ICP4), and downstream of the ICP4 gene. The 3′‐end and downstream of ICP4 gene of EHV‐1 B were found to be replaced by the corresponding region of EHV‐4, indicating that EHV‐1 B is a naturally occurring recombinant virus between progenitors of EHV‐1 P and EHV‐4. This is the first report showing a natural interspecies recombinant in alphaherpesviruses.
Diversity of Borrelia burgdorferi Sensu Lato in Russian Far East
Authors: Mediannikov Oleg Y. , Ivanov Leonid , Zdanovskaya Nina , Vorobyova Rimma , Sidelnikov Yuri , Fournier Pierre‐Edouard , Tarasevich Irina , Raoult Didier 
Published: November 2013

AbstractThirty strains of Borrelia burgdorferi sensu lato have been isolated from Ixodes persulcatus ticks and from skin lesions of Lyme disease patients in the Russian Far East from 1997 to 2003. We amplified full‐length outer surface protein A (ospA) gene of all strains. BLAST search and following phylogenetic analysis showed that strains form four well‐defined groups. Four strains belong to Borrelia afzelii species. Other strains distributed into tree major groups, identified as Borrelia garinii. Indeed, based on the ospA gene comparison, phylogenetic relationship of these groups among each other does not differ from relationship among other previously defined groups inside B. burgdorferi sensu lato genogroup, such as B. afzelii or Borrelia bissettii. Further investigations of genetic and serologic properties of the strains belonging to those groups are required in order to clarify their taxonomic status.
Molecular Basis for Adaptation of a Chimeric Dengue Type‐4/Japanese Encephalitis Virus to Vero Cells
Authors: Tang Wei‐Feng , Eshita Yuki , Tadano Masayuki , Morita Kouichi , Makino Yoshihiro 
Published: November 2013

AbstractThe premembrane and envelope (E) genes of a full‐length cDNA clone of the dengue type‐4 (DEN4) virus 814669 strain were replaced with those of the Japanese encephalitis (JE) virus JaOH0566 strain. The in vitro‐synthesized RNA transcripts prepared from chimeric cDNA were used to transfect mosquito C6/36 cells. A viable chimeric virus (designated DEN4/JE) was recovered. Unexpectedly, DEN4/JE exhibited restricted growth in Vero cells. After a serial passage in Vero cells, the Vero‐adapted chimeras were obtained (two clones, designated Strain I and Strain II, respectively). The entire genomes of DEN4/JE, Strain I, and Strain II were sequenced and compared. There were multiple mutations, but amino acid substitutions occurred only in E and nonstructural (NS) protein NS4B. Our findings in this study indicate that the 5′ nontranslated region, E, and NS4B may be involved in Vero cell adaptation in this chimeric system.
Changes in Ganglioside Content Affect the Binding of Clostridium perfringens Epsilon‐Toxin to Detergent‐Resistant Membranes of Madin‐Darby Canine Kidney Cells
Authors: Shimamoto Seiko , Tamai Eiji , Matsushita Osamu , Minami Junzaburo , Okabe Akinobu , Miyata Shigeru 
Published: November 2013

AbstractEpsilon‐toxin (ET) of Clostridium perfringens, which causes fatal enterotoxemia in ungulates, was previously shown to bind to and form a heptameric pore within the detergent‐resistant membranes (DRMs) of MDCK cells. Depletion of cholesterol has also been shown to decrease the cytotoxicity of ET and its heptamerization. In this study, we investigated the effects of changes in sphingolipids, other DRM components of MDCK cells, on the cells' susceptibility to ET. Treatment with fumonisin B1 and PDMP, inhibitors of sphingolipid and glycosphingolipid syntheses, respectively, increased the susceptibility, while D609, a sphingomyelin synthesis inhibitor, had the opposite effect. The exogenous addition of ganglioside GM1 dramatically decreased the ET binding, heptamerization and cytotoxicity. These effects were shown not to be due to ET binding to GM1 or to denaturation of ET. We also found that the ET cytotoxicity towards MDCK cells decreased with an increase in culture time. In accordance with the resistance observed for prolonged cultured cells, GM3, a major ganglioside component, increased and sialidase treatment increased their susceptibility. These results suggest that membrane‐anchored sialic acid of GM3 within DRMs inhibits ET binding, leading to prevention of the heptamerization of ET and cell death. It is also suggested that sialidase produced by this organism aids the targeting of ET to MDCK cells.
Analysis of Cellular Fatty Acids in Orientia tsutsugamushi as Taxonomic Markers
Authors: Shim Soo‐Kyoung , Shin Yong‐Kook , Choi Eun‐Na , Han Bok‐Ghee , Lee Hae‐Kyung , Choi Young‐Sill , Lee Byung‐Chul , Yoon Kwon‐Sang , Park Mi‐Yeoun 
Published: November 2013

AbstractSix Orientia strains including 3 prototype strains such as Gilliam, Karp, and Kato, and 3 strains (Boryong, Pajoo, and Yongworl) isolated in Korea, were studied for the profiles of their cellular fatty acids. All tested strains contained octadecenoic acid C18:1 ω3±3.5%), octadecanoic acid C18:0 (15.3±1.5%), and hexadecanoic acid C16:0 (12.7±1.7%) as major components; however, interestingly, eicosenoic acid C20:1 ω9c (2.6 ±0.6%) was found in all strains except the Yongworl strain. Furthermore none of the strains contained 3‐hydroxy fatty acids. The ratios of total saturated fatty acid (SFA) to total unsaturated fatty acid (UFA) were within the range of 0.34 to 0.54. These results showed that the cellular fatty acid profile should provide more reliable information for the identification of these bacteria.
Human Papillomavirus Genotypes and the p53 Codon 72 Polymorphism in Cervical Cancer of Northeastern Thailand
Authors: Settheetham‐Ishida Wannapa , Kanjanavirojkul Nipa , Kularbkaew Churairat , Ishida Takafumi 
Published: November 2013

AbstractHuman papillomavirus (HPV) infection including sub‐strain identification was studied in patients with squamous cell cervical cancer (SCCA) in Northeastern Thailand. Subjects were 90 cases of SCCA and 100 healthy controls. Prevalence of high‐risk group of HPV infection in the controls and the SCCA patients were 13.0% and 86.7%, respectively. The HPV infection significantly increased the risk for cervical cancer 43.5‐fold (95% confidential interval: 17.5–110.6; P < 0.00001). Among HPV carrier patients with SCCA (n=78), HPV‐16 was also prominent (70.5%) followed by HPV‐18 (23.1%). There was no statistical difference in the subtype distribution between the SCCA and the control groups. There was no significant association between genotype distribution of the p53 codon 72 polymorphism and HPV infection. HPV infection was confirmed as a critical risk factor for cervical cancer development in Northeast Thailand. Since polymorphism of the p53 itself as well as in combination with HPV infection may not be a genetic risk for cervical cancer, much attention should be paid to other risk factors such as sexual behavior and smoking.
Rapid Categorization of Pathogenic Escherichia coli by Multiplex PCR
Authors: Kimata Keiko , Shima Tomoko , Shimizu Miwako , Tanaka Daisuke , Isobe Junko , Gyobu Yotaku , Watahiki Masanori , Nagai Yoshiyuki 
Published: November 2013

AbstractA one‐shot multiplex polymerase chain reaction (PCR) was developed for detecting 12 virulence genes of diarrheagenic Escherichia coli. In order to differentiate between the five categories of diarrheagenic E. coli, we selected the target genes: stx1, stx2, and eaeA for enterohemorrhagic E. coli (EHEC); eaeA, bfpA, and EAF for enteropathogenic E. coli (EPEC); invE for enteroinvasive E. coli (EIEC); elt, estp, and esth for enterotoxigenic E. coli (ETEC); CVD432 and aggR for enteroaggregative E. coli (EAggEC); and astA distributed over the categories of diarrheagenic E. coli. In our multiplex PCR system, all 12 targeted genes (stx1, stx2, eaeA, invE, elt, estp, astA, esth, bfpA, aggR, EAF, and CVD432) were amplified in a single PCR reaction in one tube and detected by electrophoresis. Using our multiplex PCR, the 208 clinically isolated strains of diarrheagenic E. coli in our laboratory were successfully categorized and easily analyzed for the presence of virulence plasmids.
CCR7 Ligand‐Enhanced Phagocytosis of Various Antigens in Mature Dendritic Cells—Time Course and Antigen Distribution Different from Phagocytosis in Immature Dendritic Cells
Authors: Kikuchi Kazuhiro , Yanagawa Yoshiki , Onoé Kazunori 
Published: November 2013

AbstractCC chemokine receptor 7 (CCR7) is selectively expressed on mature dendritic cells (DC). The CCR7 ligands, CC chemokine ligand (CCL) 19 and CCL21, facilitate migration of mature DC from the peripheral tissues to regional lymph nodes. We previously demonstrated that CCR7 ligands induced rapid receptor‐mediated endocytosis of dextran in mature DC. In the present study, we further examined the effects of CCR7 ligands on endocytosis of other kinds of antigen, mannosilated bovine serum albumin (Mann‐BSA), Escherichia coli (E. coli), or ovalbumin‐containing immune complex (OVA‐IC), by mature DC. We found that CCR7 ligands enhanced the endocytosis of Mann BSA, E. coli, and OVA‐IC in mature DC but not in immature DC. The endocytosis of BSA was not enhanced by CCR7 ligands. Furthermore, the phagocytosis of OVA‐IC was significantly inhibited by anti‐FcY receptor III/II antibody. These results demonstrate that CCR7 ligands enhance only receptor‐mediated endocytosis by mature DC. When rapidly phagocytosed E. coli were traced in CCL19‐treated mature DC, most of the phagocytosed E. coli did not colocalize with the lysosomal marker: lysosome‐associated membrane protein‐1 (Lamp‐1), whereas most of E. coli taken up relatively slowly by immature DC colocalized with Lamp‐1. These results suggest that phagocytosis of antigens by immature and mature DC plays different functional roles.
Rapid and Simple Identification of Orientia tsutsugamushi from Other Group Rickettsiae by Duplex PCR Assay Using groEL Gene
Authors: Park Hyo‐Soon , Lee Jung‐Hee , Jeong Eun‐Ju , Kim Jung‐Eun , Hong Sung‐Jin , Park Tae‐Kyu , Kim Tae‐Yung , Jang Won‐Jong , Park Kyung‐Hee , Kim Bum‐Joon , Kook Yoon‐Hoh , Lee Seung‐Hyun 
Published: November 2013

AbstractIn this study, two new duplex PCR methods based on the groEL gene were developed and investigated for the diagnosis of rickettsiae. The first duplex PCR assay amplified the 229‐bp and the 366‐bp DNAs of 6 strains including typhus group (TG) and spotted fever group (SFG) rickettsiae, and 5 scrub typhus group (STG) rickettsiae, respectively. The second duplex PCR assay amplified the 397‐bp and the 213‐bp DNAs of 6 Rickettsia strains and 5 STG strains. These duplex PCR methods could simultaneously perform the rapid identification of rickettsiae and the differential diagnosis of STG and other group rickettsiae in a single reaction.
Chemokine and Cytokine Production in Susceptible C3H/HeN Mice and Resistant BALB/c Mice during Orientia tsutsugamushi Infection
Authors: Yun Ji‐Hyun , Koh Young‐Sang , Lee Keun‐Hwa , Hyun Jin‐Won , Choi Yeon‐Joo , Jang Won‐Jong , Park Kyung‐Hee , Cho Nam‐Hyuk , Seong Seung‐Yong , Choi Myung‐Sik , Kim Ik‐Sang 
Published: November 2013

AbstractTo understand the pathogenesis of scrub typhus, we examined chemokine and cytokine production in susceptible (C3H/HeN) and resistant (BALB/c) mice after infection with O. tsutsugamushi Gilliam. C3H/HeN mice produced high levels of chemokines macrophage inflammatory proteins 1α (MIP‐1α), MIP‐2, monocyte chemoattractant protein 1 (MCP‐1), and cytokines gamma‐interferon (IFN‐γ), interleukin‐12 (IL‐12), IL‐10, and tumor necrosis factor alpha (TNF‐α) in response to O. tsutsugamushi infection, compared to BALB/c mice. Chemokine profiles in infected mice correlated well with the kinetics of inflammatory cell infiltration. Hyperproduction of chemokines and cytokines was observed in another susceptible‐infection model (BALB/c‐Karp). These results suggest that hyperproduction of chemokines and cytokines are associated with susceptibility during O. tsutsugamushi infection.
Homologous Recombination with Linear DNA to Insert Antigenic Protein in the Flagellin: Improvement of the Th1 Immune Response
Authors: Moigne Vincent Le , Robreau Georges , Mahana Wahib 
Published: November 2013

AbstractBacterial flagellin is a surface protein with numerous advantages for the presentation of exogenous peptides. However, the production of recombinant bacteria and the expression of fusion proteins is laborious and time consuming. Here, we present a simple way to produce modified bacteria. Partially deleted, non‐functional, chromosomal flagellin gene (fliC) was changed using homologous recombination by a functional linear fliC gene in which we introduced an exogenous oligonucleotide encoding for the peptide of interest. The modified fliC gene was produced by polymerase chain amplification. Linear amplicons were introduced into the non‐motile E. coli by electroporation. The formation of functional flagellar filaments allowed the discrimination of motile transformants from non‐motile, non‐transformed cells. Thus antibiotic selection and gene expression inductors are not required since transformed bacteria can be easily isolated and used as a vector and adjuvant for immunization. To validate this hypothesis, we studied the immune response against the N‐terminal peptide of Clostridium tyrobutyricum flagellin fragment. BALB/c mice were immunized either with the protein displayed as flagellin fusion protein on the surface of E. coli, with the recombinant protein in Freund's adjuvant (FA), or with the pcDNA3 vector bearing the DNA fragment encoding this protein. Immunization with the flagellin recombinant bacteria induced a strong Th1 response as measured by high level of IFN‐γ production and the lack of IL‐4 production. The results indicate that the flagellar filament protein carrying a specific epitope can be a potent inducer of the Th1 cellular response.
Capsule of Streptococcus pyogenes Is Essential for Delayed Death of Mice in a Model of Streptococcal Toxic Shock Syndrome
Authors: Iida Ken‐ichiro , Seki Masanori , Saito Mitsumasa , Kawamura Yoshiaki , Kajiwara Hideko , Yoshida Shin‐ichi 
Published: November 2013

AbstractWe have previously reported a mouse model of severe group A streptococcal infection (Microbiol. Immunol. 45: 777–786, 2001). When we injected Streptococcus pyogenes strains intramuscularly, the mice suffered from acute phase of infection for a few days but recovered from the illness and gained body weight. These mice, however, began to die after 3 weeks of infection, which we called ‘delayed death.’ Bacterial strains isolated from organs of the dead mice showed thick capsules. We, therefore, constructed a hyaluronic acid capsule gene, hasA, knockout mutant by homologous recombination and the effect of capsule on the death was observed. hasA knockout strain did not cause delayed death, though it caused acute death at high doses of infection. According to this result, the capsule is a critical pathogenic factor for causing the delayed death in our mouse model.
Adjuvant Effect of the Propionibacterium acnes and Its Purified Soluble Polysaccharide on the Immunization with Plasmidial DNA Containing a Trypanosoma cruzi Gene
Authors: Mussalem Juliana Sekeres , Vasconcelos José Ronnie C. , Squaiella Carla Cristina , Ananias Renata Zeigler , Braga Eleni Gonçalves , Rodrigues Maurício M. , Longo‐Maugéri Ieda Maria 
Published: November 2013

AbstractIn the present work we investigated the role of killed Propionibacterium acnes or a soluble polysaccharide extracted from bacterium cell wall in modulated experimental immunization with plasmidial DNA. We used a plasmid, p154/13, containing a gene‐encoding catalytic domain of Trypanosoma cruzi (T. cruzi) trans‐sialidase. As previously described, immunization of BALB/c mice with p154/13 elicited humoral, cell‐mediated and protective immune responses against T. cruzi infection. In this study we describe that both P. acnes and its soluble polysaccharide fraction have the ability to modulate the immune response elicited by p154/13. Treatment with these adjuvants enhanced specific trans‐sialidase Th1 immune response, as revealed by a lower IgG1/IgG2a ratio and stronger in vitro IFN‐γ synthesis by CD4+ T cells. The most important fact was that treatment with P. acnes or its soluble polysaccharide fraction in the presence of p154/13 significantly reduced the peak of parasitemia observed 7 to 8 days after T. cruzi challenge. These data suggest that P. acnes or its soluble polysaccharide fraction may improve the protective potential of a DNA vaccine against experimental T. cruzi infection.
Analysis of Amphotericin B‐Induced Cell Signaling with Chemical Inhibitors of Signaling Molecules
Authors: Matsuo Kenichiro , Hotokezaka Hitoshi , Ohara Naoya , Fujimura Yuji , Yoshimura Atsutoshi , Okada Yukio , Hara Yoshitaka , Yoshida Noriaki , Nakayama Koji 
Published: November 2013

AbstractAlthough amphotericin B (AmB) is a major polyene antibiotic against invasive fungal infection, administration to patients sometimes causes inflammatory side effects, which limits the usage of the antibiotic. We studied the intracellular signaling that was induced by AmB. p65 (RelA) of nuclear factor‐κB (NF‐κB), a well‐known signaling molecule as an inducer of proinflammatory cytokines, was phosphorylated by AmB in RAW264.7 cells, a monocyte‐like cell line. Among chemical inhibitors of signaling molecules, U‐73122 (phospholipase C (PLC) inhibitor), Gö6976 (protein kinase C (PKC) inhibitor), BAPTA‐AM (calcium chelator), LFM‐A13 (Bruton's tyrosine kinase (Btk)‐specific inhibitor), and PP2 (c‐Src kinase inhibitor) suppressed AmB‐induced phosphorylation of p65 and translocation of p65 into the nucleus. U‐73122 and Gö6976 reduced AmB‐mediated induction of proinflammatory cytokines (tumor necrosis factor (TNF)‐α and interleukin (IL)‐6) in RAW264.7 cells. Furthermore, AmB‐induced activation of NF‐κB was observed in toll‐like receptor (TLR) 2‐expressed cells, and the activation of NF‐κB was inhibited by U‐73122, whereas peptidoglycan‐induced NF‐κB activation, which was also dependent on TLR2, was not inhibited by U‐73122. Finally, U‐73122 partially suppressed in vivo production of TNF‐α and IL‐6 induced by AmB administration in BALB/c mice. These results suggested that the signaling from AmB stimulation to proinflammatory cytokine production is mediated by TLR2, Btk, PLC, PKC, c‐Src and NF‐κB. These signaling molecules may become a target for chemotherapy suppressing AmB‐induced proinflammatory cytokine production.
Analysis of a Bacterial Lipopolysaccharide‐Activated Serine Kinase That Phosphorylates p65/L‐Plastin in Macrophages
Authors: Hagi Akifumi , Hirata Hajime , Shinomiya Hiroto 
Published: November 2013

AbstractWe previously identified p65/L‐plastin as a phosphorylated protein in LPS‐stimulated macrophages and determined its phosphorylation site. In vitro kinase assay using peptide substrates revealed that LPS‐stimulated kinase activity selectively phosphorylated their serine‐5 (Ser‐5) residue. Kinase inhibitors for cAMP‐dependent kinase such as H‐89 inhibited the Ser‐5 phosphorylation, but cAMP was not essential for the kinase activity. The LPS‐stimulated kinase activity in cytosol fractions of macrophages was recovered as a sharp peak by anion exchange chromatography. These findings suggest that an as yet unknown H‐89‐sensitive serine kinase is rapidly activated by LPS stimulation and then phosphorylates p65/L‐plastin, playing a vital role in macrophage activation.