bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of the Coxiella Sp. Detected from Haemaphysalis longicornis Ticks in Korea
Authors: Lee Jung‐Hee , Park Hyo‐Soon , Jang Won‐Jong , Koh Seong‐Eun , Park Tae‐Kyu , Kang Shin‐Seok , Kim Bum‐Joon , Kook Yoon‐Hoh , Park Kyung‐Hee , Lee Seung‐Hyun 
Published: November 2013

AbstractTwo Haemaphysalis longicornis ticks were found positive in PCR assay of com‐1 gene to detect Coxiella burnetii DNA from 100 ticks. The nucleotide sequences of com‐1 and 16S rRNA gene were determined from 2 ticks and compared with those of other C. burnetii strains. The results suggest that H. longicornis harbor Coxiella sp. bacteria in Korea. Furthermore, icd, cbhE′, and cbbE′ genes are C. burnetii specific genes whereas com‐1 gene is Coxiella genus specific gene. This study gives the first documentation to prove the existence of Coxiella sp. in tick collected in Korea.
Preparation of Virosomes Coated with the Vesicular Stomatitis Virus Glycoprotein as Efficient Gene Transfer Vehicles for Animal Cells
Authors: Shoji Jun'ichi , Tanihara Yuko , Uchiyama Tsuneo , Kawai Akihiko 
Published: November 2013

AbstractThe vesicular stomatitis virus (VSV) glycoprotein (G) was used to prepare virosomes as a model vehicle of gene transfer to animal cells, for which viral envelope functions (receptor recognition and binding and the pH‐dependent membrane‐fusion) were expected to work. Plasmid DNA (pEGFP‐N1; Clontech) was first encapsulated into liposomes by a method of repeated freezing and thawing of the mixture of DNA and lipids (phosphatidylcholine, phosphatidylserine and cholesterol mixed at a molar ratio of 5:1:4). Then, particle size of the liposomes was stepwise reduced to 200 nm or less in diameter by successive filtrations through a series of plastic filters of various pore sizes (10 μm, 2 μm, 0.65 μm, and then 0.45 μm). Assembly of the VSV G protein‐coated liposomes (VSV G‐virosomes) was performed by mixing the DNA‐encapsulated liposome suspensions with the purified VSV G proteins at pH 5.5, followed by ultracentrifugation in a discontinuous sucrose gradient. The highest gene‐transducing activity was detected in a single band formed between 20% and 45% sucrose layers. Negatively stained electron microscopic images showed that the band contained spherical particles of various sizes, ranging from 40 to 140 nm in diameter, that were covered with viral spike projections. The VSV G‐virosomes displayed a roughly similar level of gene‐transducing activity to that mediated by cationic liposomes (e.g., Lipofectamine), which was blocked either by pretreatment with anti‐VSV G antiserum or by addition of 20 MM NH4Cl to transfected cultures. From these results, we assume that the virosome‐mediated gene‐transduction was first achieved by using the whole functions of VSV G protein, and can also be used for further studies of the protein.
Human Cytomegalovirus (HCMV) Infection in Osteosarcoma Cell Line Suppresses GM‐CSF Production by Induction of TGF‐β
Authors: Kwon Ye Jin , Kim Dae‐Joong , Kim Jung Heon , Park Chung‐Gyu , Cha Chang‐Yong , Hwang Eung‐Soo 
Published: November 2013

AbstractThis study was performed to elucidate the possible mechanism of the disturbance of hemopoiesis by HCMV infection. Saos‐2 cells constitutively express mRNA of GM‐CSF, and its expression was profoundly decreased by HCMV infection, which required full replication of the virus and was mediated by soluble factors released from the HCMV‐infected Saos‐2 cells. TGF‐β1 production was statistically and significantly increased from one day after HCMV infection. Expression and production of GM‐CSF in Saos‐2 cells were restored when a culture supernatant of HCMV‐infected Saos‐2 cells was reacted with neutralizing anti‐TGF‐β antibody. Conclusively, HCMV inhibits GM‐CSF expression in Saos‐2 cells partly by the increased production of TGF‐β1.
Immunomagnetic Capture RT‐PCR for Detection of Norovirus from Foods Implicated in a Foodborne Outbreak
Authors: Kobayashi Shinichi , Natori Katsuro , Takeda Naokazu , Sakae Kenji 
Published: November 2013

AbstractIn February 2001, an outbreak of acute gastroenteritis due to Norovirus (NV) occurred among employees of 11 companies in Aichi Prefecture. The illness was strongly associated with eating a delivered box‐lunch. The use of magnetic beads coated with the antibody to the baculovirus‐expressed recombinant capsid proteins of the Chiba virus (rCV) facilitated capture of NV from the food items implicated in the outbreak. Following immunomagnetic capture, NV bound to the beads was detected by reverse transcription‐polymerase chain reaction (RT‐PCR). Of the nine food items tested, two were positive for a genogroup 1 NV. Sequence analysis of RT‐PCR products indicated that the nucleotide sequences of NV strains from foods were almost identical to those of NV strains detected in stool samples of ill patients. As the immunocapture RT‐PCR method is simple and easy to perform, this technique should be useful for the detection of NV from outbreak‐implicated foods.
Genetic Typing of Feline Rabies Virus Isolated in Greater Bangkok, Thailand
Authors: Kasempimolporn Songsri , Saengseesom Wachiraporn , Tirawatnapong Thaweesak , Puempumpanich Sununta , Sitprija Visith 
Published: November 2013

AbstractTo study the molecular epidemiology of rabies virus that is prevalent among cats in greater Bangkok, Thailand, a total of 17 rabies virus isolates from cats were characterized and compared with 120 rabies virus isolates from dogs. Analyses were performed on the genetic polymorphism in the rabies virus nucleoprotein (N) gene. Rabies virus N gene of isolates was amplified by reverse transcription‐polymerase chain reaction. The diversity of N gene was revealed by the restriction fragment length polymorphism (RFLP) method. The rabies virus isolates from cats could be classified into 5 types, designated as Dd I‐Hf I, Dd II‐Hf II, Dd III‐Hf I, Dd IV‐Hf I, and Dd IV‐Hf III. Type Dd I‐Hf I was encountered more frequently than the others. It was apparent that no less than five rabies virus types presented in the areas of Bangkok. Moreover, all five RFLP patterns were typical of those which had been observed in dogs. Our findings suggest that there had been viral transmission between the dogs and the cats.
Cytotoxicity and Enterotoxicity of the Thermostable Direct Hemolysin‐Deletion Mutants of Vibrio parahaemolyticus
Authors: Park Kwon‐Sam , Ono Takahiro , Rokuda Mitsuhiro , Jang Myoung‐Ho , Iida Tetsuya , Honda Takeshi 
Published: November 2013

AbstractThe thermostable direct hemolysin (TDH) has been proposed to be a major virulence factor of Vibrio parahaemolyticus. We have recently completed the genome sequence of a TDH‐producing V. parahaemolyticus strain, RIMD2210633. In this study, we constructed tdh‐deletion mutants from the sequenced strain by homologous recombination and analyzed their phenotypes. Although the deletion of both copies of tdh completely abolished the hemolytic activity of the wild‐type strain, the deletion did not affect the cytotoxicity to HeLa cells. Enterotoxicity, assayed by the rabbit ileal loop test, was lowered by tdh deletion, but the mutant still showed partial fluid accumulation in rabbit intestine. These results indicate that the cytotoxicity and enterotoxicity of TDH‐producing V. parahaemolyticus are not explained by TDH alone, and suggest that an unknown virulence factor(s) could be involved in these pathogenic activities.
Modulation of the Immune System by Listeria monocytogenes‐Mediated Gene Transfer into Mammalian Cells
Authors: Shen Hua , Kanoh Makoto , Liu Fengzhi , Maruyama Saho , Asano Yoshihiro 
Published: November 2013

AbstractIn this study, we established a method for Listeria monocytogenes (Lm)‐mediated gene transfer into mammalian cells to manipulate the immune response of the host during infection by pathogens. We used the Lm‐mediated gene transfer method in an in vivo study to manipulate host immune responses against Leishmania major (L. major)‐infection. The injection of Lm modulated the susceptible host into a resistant state against L. major‐infection. A more efficient protective effect was obtained with the injection of IL‐12‐cDNA containing Lm, and the protective effect was stronger than that of the resistant strain. The protective mechanism of Lm‐injection against L. major‐infection observed here appeared to be a result of the activation of the local immune system by the Lm‐mediated gene transfer method. The present study is the first demonstration that a gene introduced into a host by Lm works to modulate the murine host immune response against infections in vivo. Since this system strongly induces Th1 responses and suppresses Th2 responses in infected hosts, the system can be used for controlling infectious diseases and for protection against allergic responses in the future.
A Major Li+ Extrusion System NhaB of Pseudomonas aeruginosa: Comparison with the Major Na+ Extrusion System NhaP
Authors: Kuroda Teruo , Fujita Naoko , Utsugi Junko , Kuroda Masayuki , Mizushima Tohru , Tsuchiya Tomofusa 
Published: November 2013

AbstractA gene encoding a Li+ extrusion system was cloned from the chromosomal DNA of Pseudomonas aeruginosa and expressed in Escherichia coli cells. The gene enabled growth of E. coli KNabc cells, which were unable to grow in the presence of 10 MM LiCl or 0.1 M NaCl because of the lack of major Na+(Li+)/H+ antiporters. We detected Li+/H+ and Na+/H+ antiport activities in membrane vesicles prepared from E. coli KNabc cells that harbored a plasmid carrying the cloned gene. Activity of this antiporter was pH‐dependent with an optimal pH activity between pH 7.5 and 8.5. These properties indicate that this antiporter is different from NhaP, an Na+/H+ antiporter from P. aeruginosa that we reported previously, and that is rather specific to Na+ but it cannot extrude Li+ effectively. The gene was sequenced and an open reading frame (ORF) was identified. The amino acid sequence deduced from the ORF showed homology (about 60% identity and 90% similarity) with that of the NhaB Na+/H+ antiporters of E. coli and Vibrio parahemolyticus. Thus, we designated the antiporter as NhaB of P. aeruginosa. E. coli KNabc carrying the nhaB gene from P. aeruginosa was able to grow in the presence of 10 to 50 MM LiCl, although KNabc carrying nhaP was unable to grow in these conditions. The antiport activity of NhaB from P. aeruginosa was produced in E. coli and showed apparent Km values for Li+ and Na+ of 2.0 MM and 1.3 MM, respectively. The antiport activity was inhibited by amiloride with a Ki value for Li+ and Na+ of 0.03 MM and 0.04 MM, respectively.
Antibacterial Activity of Hydrolyzable Tannins Derived from Medicinal Plants against Helicobacter pylori
Authors: Funatogawa Keiji , Hayashi Shunji , Shimomura Hirofumi , Yoshida Takashi , Hatano Tsutomu , Ito Hideyuki , Hirai Yoshikazu 
Published: November 2013

AbstractHelicobacter pylori is a major etiological agent in gastroduodenal disorders. In this study, we isolated 36 polyphenols and 4 terpenoids from medicinal plants, and investigated their antibacterial activity against H. pylori in vitro. All hydrolyzable tannins tested demonstrated promising antibacterial activity against H. pylori. Monomeric hydrolyzable tannins revealed especially strong activity. Other compounds demonstrated minimal antibacterial activity with a few exceptions. A monomeric hydrolyzable tannin, Tellimagrandin I demonstrated time‐ and dose‐dependent bactericidal activity against H. pylori in vitro. On the other hand, hydrolyzable tannins did not affect the viability of MKN‐28 cells derived from human gastric epithelium. Hydrolyzable tannins, therefore, have potential as new and safe therapeutic regimens against H. pylori infection. Furthermore, we investigated effects of hydrolyzable tannins on lipid bilayer membranes. All the hydrolyzable tannins tested demonstrated dose‐dependent membrane‐damaging activity. However, it remains to be elucidated whether their membrane‐damaging activity directly contributes to their antibacterial action.
Infection of Different Cell Lines of Neural Origin with Subacute Sclerosing Panencephalitis (SSPE) Virus
Authors: Ishida Hiroshi , Ayata Minoru , Shingai Masashi , Matsunaga Isamu , Seto Yoshiyuki , Katayama Yuko , Iritani Nobuhiro , Seya Tsukasa , Yanagi Yusuke , Matsuoka Osamu , Yamano Tsunekazu , Ogura Hisashi 
Published: November 2013

AbstractMeasles virus is the causative agent of subacute sclerosing panencephalitis (SSPE). The viruses isolated from brain cells of patients with SSPE (called SSPE viruses) are defective in cell‐free virus production in vitro. To investigate the cell tropism of three strains of SSPE virus (Osaka‐1, Osaka‐2, Osaka‐3), SSPE virus‐infected cell cultures were treated with cytochalasin D to prepare virus‐like particles (CD‐VLPs). All CD‐VLPs formed syncytia after infection in CHO cells expressing CD150 but not in those expressing CD46. In addition, an antibody to CD46 did not block the infection of Vero cells by SSPE CD‐VLPs. The results were consistent with our previous suggestion that one or more unidentified receptors might be involved in the entry process. Infection with the CD‐VLPs from three SSPE strains was further examined in different human cell lines, including those of neural origin, and was found to induce syncytia in epithelial cells (HeLa and 293T) as well as neuroblastoma cells (IMR‐32 and SK‐N‐SH) with varying efficiency. SSPE CD‐VLPs also infected glioblastoma cells (A172) and astrocytoma cells (U‐251) but syncytial formation was rarely induced. These epithelial and neural cell lines were not permissive for the replication of wild‐type MV. Together with our previous observations, these results suggest that the cell entry receptor is the major factor determining the cell tropism of SSPE viruses. Further studies are necessary to identify other viral and/or cellular factors that might be involved in the replication of SSPE virus in specific neural cells and in the brain.
Identification of β‐Tubulin Isoform V as an Autoantigen in Allergic Rhinitis by a Proteomic Approach
Authors: Nakamura Manabu , Tsutsumi Kouichiro , Ooka Seido , Sekine Taichi , Koizuka Izumi , Nishioka Kusuki , Kato Tomohiro 
Published: November 2013

AbstractAutoantibodies to IgE and p2‐adrenergic receptor have been reported in patients with allergic rhinitis. To investigate whether autoimmunity in allergic rhinitis is directed to such limited molecules or directed to a wide range of self proteins, we here attempted to survey autoantigens/autoantibodies comprehensively, using proteomics. Specifically, we separated proteins extracted from peripheral blood mononuclear cells by 2‐dimensional electrophoresis and then detected autoantigens by subsequent western blotting with sera from patients with allergic rhinitis. As a result, we detected multiple autoantigens, some of which were further identified by mass fingerprinting. Next, we confirmed antigenicity of one of the identified autoantigens, β‐tubulin isoform V (β‐tubV), using a recombinant protein and then measured prevalence of the anti‐β‐tubV autoantibodies. As a result, 52% of the tested patients with allergic rhinitis were found to possess anti‐β‐tubV autoantibodies. Our study indicates that autoimmunity is a common phenomena and β‐tubV is one of the major autoantigens in allergic rhinitis.
A Quorum‐Sensing Autoinducer Enhances the mexAB‐oprM Efflux‐Pump Expression without the MexR‐Mediated Regulation in Pseudomonas aeruginosa
Authors: Sawada Isao , Maseda Hideaki , Nakae Taiji , Uchiyama Hiroo , Nomura Nobuhiko 
Published: November 2013

AbstractWe have previously described that the quorum‐sensing autoinducer, N‐butyryl‐L‐homoserine lactone (C4‐HSL) enhances mexAB‐oprM expression, and this C4‐HSL‐mediated enhancement of mexAB‐oprM expression was repressed by MexT, a positive regulator of the mexEF‐oprN operon. In this study, we investigated the interaction between C4‐HSL and mexR by using a knockout mutant. It was indicated that the C4‐HSL‐mediated enhancement of mexAB‐oprM expression occurred without MexR‐mediated regulation. Furthermore, it was observed that the C4‐HSL‐mediated enhancement of mexAB‐oprM expression without the MexR‐mediated regulation was repressed by MexT.
Comparison of Five PCR Assays for Detecting Chlamydophila pneumoniae DNA
Authors: Fukano Hiroshi 
Published: November 2013

AbstractWe compared five different polymerase chain reaction (PCR) assays for the detection of Chlamydophila pneumoniae DNA using highly purified elementary bodies (EBs) and peripheral blood mononuclear cells (PBMCs) from healthy blood donors. The primers were as follows; two targeting the 16S rRNA gene, one targeting the ompA gene, one targeting the Pst‐I gene, and one targeting the 53 kDa outer membrane protein gene. The 16S rRNA touchdown enzyme time release (TETR) PCR, the ompA nested PCR and the 53 kDa nested PCR were the most sensitive assays and could detect one or more EB per assay. These three PCRs also had the same reproducibility, but the minimal amount of C. pneumoniae that could be reproducibly detected (10 of 10 testing positive) was 20 EBs. In a sample of specimens from healthy blood donors, we found 5 of 77 (6.5%) PBMCs specimens to have C. pneumoniae DNA according to the nested ompA PCR. Specimens with the 16S rRNA TETR and 53 kDa nested assays were found to have C. pneumoniae DNA 7 of 77 (9.1%) and 18 of 77 (23.4%) specimens, respectively. The other two assays failed to detect even a single positive. However, the detection rate decreased with repeated testing of the same samples. Our newly designed 53 kDa nested PCR may be as useful as the other four recommended PCR assays and may be a more useful assay for the detection of C. pneumoniae DNA from PBMCs.
Application of In Vitro Mutagenesis to Identify the Gene Responsible for Cold Agglutination Phenotype of Streptococcus mutans
Authors: Sato Yutaka , Okamoto Kazuko , Kagami Akiyoshi , Yamamoto Yasuhito , Ohta Kazumasa , Igarashi Takeshi , Kizaki Harutoshi 
Published: November 2013

AbstractA previously unidentified protein with an apparent molecular mass of 120 kDa was detected in some Streptococcus mutans strains including the natural isolate strain Z1. This protein was likely involved in the cold‐agglutination of the strain, since a correlation between this phenotype and expression of the 120 kDa protein was found. We have applied random mutagenesis by in vitro transposition with the Himar1 minitransposon and isolated three cold‐agglutination‐negative mutants of this strain from approximately 2,000 mutants screened. A 2.5 kb chromosomal fragment flanking the minitransposon in one of the three mutants was amplified by PCR‐based chromosome walking and the minitransposon insertion in the other two mutants occurred also within the same region. Nucleotide sequencing of the region revealed a 1617 nt open reading frame specifying a putative protein of 538 amino acid residues with a calculated molecular weight of 57,192. The deduced eight amino acid sequence following a putative signal sequence completely coincided with the N‐terminal octapeptide sequence of the 120 kDa protein determined by the Edman degradation. Therefore, the 1617 nt gene unexpectedly encoded the 120 kDa protein from S. mutans. Interestingly, this gene encoded a collagen adhesin homologue. In vitro mutagenesis using the Himar1 minitransposon was successfully applied to S. mutans.
New World Relapsing Fever Borrelia Found in Ornithodoros porcinus Ticks in Central Tanzania
Authors: Mitani Harumi , Talbert Alison , Fukunaga Masahito 
Published: November 2013

AbstractTicks were collected from 8 houses in Mvumi Mission village, near Dodoma, Tanzania. All ticks were examined for Borrelia infestation by flagellin gene‐based nested polymerase chain reaction. All houses were highly infested with ticks, and all ticks collected were of the Ornithodoros porcinus species. Fifty‐one out of 120 ticks were infected with spirochetes, and a flagellin gene sequence comparison showed that most of the spirochetes belonged to Borrelia duttonii, which is the causative agent of tick‐borne relapsing fever in East Africa. The rest of the spirochetes were quite different from B. duttonii and instead resembled the New World tick‐borne relapsing fever borreliae. Phylogenetic analysis using 16S ribosomal RNA gene sequences also supported the interpretation that the spirochete was a Borrelia species distinct from previously described members of the genus.
Distribution of Virulence‐Associated Genes in Vibrio mimicus Isolates from Clinical and Environmental Origins
Authors: Shinoda Sumio , Nakagawa Tomoko , Shi Lei , Bi Kewei , Kanoh Yasuko , Tomochika Ken‐ichi , Miyoshi Shin‐ichi , Shimada Toshio 
Published: November 2013

AbstractDistribution of virulence‐associated genes in Vibrio mimicus was studied including the toxin genes ctxA, tdh, st and vmh and the genes necessary for regulation of toxin production, toxR, toxS, toxT, tcpA and tcpP. Approximately half of clinical V. mimicus isolates possessed one or more genes encoding V. cholerae enterotoxic factors such as ctxA, tdh and st. All of the clinical and environmental isolates possessed vmh encoding V. mimicus hemolysin (VMH). The ctxA encoding cholera toxin was detected in only 2 strains, 5% of the clinical isolates. Furthermore, there were very few strains possessing tcpP and toxT needed for the expression of ctxA. These results may suggest that VMH is a more important pathogenic factor than well recognized toxins such as cholera toxin (CT) in V. mimicus infection.
A New Yeast, Malassezia yamatoensis, Isolated from a Patient with Seborrheic Dermatitis, and Its Distribution in Patients and Healthy Subjects
Authors: Sugita Takashi , Tajima Mami , Takashima Masako , Amaya Misato , Saito Masuyoshi , Tsuboi Ryoji , Nishikawa Akemi 
Published: November 2013

AbstractOver the last few years, new Malassezia species have been found regularly in Japanese subjects. We isolated another new Malassezia species from a Japanese patient with seborrheic dermatitis (SD), and named it M. yamatoensis. In its physiological characteristics and the utilization of Tween by M. yamatoensis is similar to that of M. furfur and M. dermatis. It is distinguished by its growth temperature. To examine the distribution of the microorganism in the skin of patients with SD and atopic dermatitis (AD), and healthy subjects, we applied transparent dressings to the skin, and detected M. yamatoensis DNA using a non‐culture‐based method that consisted of nested PCR with specific primers. M. yamatoensis DNA was detected from 3 of 31 SD patients (9.7%), 5 of 36 AD patients (13.9%), and 1 of 22 healthy subjects (4.6%). Therefore, M. yamatoensis is a rare member of the cutaneous microflora.
Identification and Characterization of the pswP Gene Required for the Parallel Production of Prodigiosin and Serrawettin W1 in Serratia marcescens
Authors: Sunaga Shinyu , Li Hong , Sato Yohei , Nakagawa Yoji , Matsuyama Tohey 
Published: November 2013

AbstractSerratia marcescens mutants defective in production of the red pigment prodigiosin and the biosurfactant serrawettin W1 in parallel were isolated by transposon mutagenesis of strain 274. Cloning of the DNA fragment required for production of these secondary metabolites with different chemical structures pointed out a novel open reading frame (ORF) named pswP. The putative product PswP (230 aa) has the distinct signature sequence consensus among members of phosphopantetheinyl transferase (PPTase) which phosphopantetheinylates peptidyl carrier protein (PCP) mostly integrated in the nonribosomal peptide synthetases (NRPSs) system. Since serrawettin W1 belongs to the cyclodepsipeptides, which are biosynthesized through the NRPSs system, and one pyrrole ring in prodigiosin has been reported as a derivative of L‐proline tethered to phosphopantetheinylated PCP, the mutation in the single gene pswP seems responsible for parallel failure in production of prodigiosin and serrawettin W1.