bluuebunny/crossref_metadata_2025_split huggingface dataset:
Growth and Virulence Alterations of Equine Herpesvirus 1 by Insertion of a Green Fluorescent Protein Gene in the Intergenic Region between ORFs 62 and 63
Authors: Ibrahim El Sayed Moustafa , Pagmajav Ochir , Yamaguchi Tsuyoshi , Matsumura Tomio , Fukushi Hideto 
Published: November 2013

AbstractNucleotide sequences of the intergenic region between ORF 62 and ORF 63 of equine herpesvirus 1 (EHV‐1) isolates were analyzed. The sequences of this region consisted of variable and conserved domains among EHV‐1 isolates. An EHV‐1 mutant, Ab4‐GFP, was constructed by inserting a green fluorescent protein (GFP) expression cassette flanked by loxP at both ends into the intergenic region between ORF 62 and ORF 63. Another mutant, Ab4‐loxP, which contains one loxP site, was constructed by excision of the GFP cassette from the Ab4‐GFP virus genome by cre enzyme. The recombinant Ab4‐GFP formed smaller plaques than the wild type in MDBK cells. Virus production also decreased for Ab4‐GFP in multi‐step growth analyses. Virulence of Ab4‐GFP in both mice and hamsters was weaker than that of the wild type. Ab4‐loxP exhibited properties similar to those of the wild type. These results suggest that the intergenic region between ORF 62 and ORF 63 plays various roles in the virus growth.
Stimulation of Phagocytosis of Influenza Virus‐Infected Cells through Surface Desialylation of Macrophages by Viral Neuraminidase
Authors: Watanabe Yuichi , Shiratsuchi Akiko , Shimizu Kazufumi , Takizawa Takenori , Nakanishi Yoshinobu 
Published: November 2013

AbstractCells infected with influenza A virus undergo apoptosis and become susceptible to phosphatidylserine‐mediated phagocytosis by macrophages. This study was undertaken to elucidate the mechanism underlying our previous finding that the activity of viral neuraminidase (NA) is required for efficient phagocytosis. Treatment of macrophages, not influenza virus‐infected cells, with Arthrobacter ureafaciens NA or virus‐infected cells expressing viral NA augmented the level of phagocytosis of virus‐infected cells but not of latex beads or cells undergoing Fas‐induced apoptosis. Oligosaccharides, including sialyllactose, bound to influenza virus‐infected cells and inhibited phagocytosis by macrophages. These results indicate that surface desialylation of macrophages by influenza virus NA modulates the mode of association between macrophages and target virus‐infected cells and stimulates phosphatidylserine‐mediated phagocytosis.
Yersinia enterocolitica Serovar O:8 Infection in Breeding Monkeys in Japan
Authors: Iwata Taketoshi , Une Yumi , Okatani Alexandre Tomomitsu , Kaneko Sei‐ichi , Namai Satoshi , Yoshida Shin‐ichiro , Horisaka Tomoko , Horikita Tetsuya , Nakadai Aya , Hayashidani Hideki 
Published: November 2013

AbstractIn the period from December 2002 to January 2003, 5 of 50 squirrel monkeys (Saimiri sciureus) housed at a Zoological Garden in the Kanto region of Japan died following a few days' history of diarrhea. After this outbreak had ended in the squirrel monkeys, 1 of 2 dark‐handed gibbons (Hylobates agilis) died in April of 2003, showing similar clinical signs. We examined the organs of 3 of the dead squirrel monkeys and of the dark‐handed gibbon, and Yersinia enterocolitica serovar O:8, which is the most pathogenic serovar of Y. enterocolitica, was isolated. In order to determine the source and the transmission route of infection, 98 fecal samples (45 from squirrel monkeys, 20 from other monkeys of 18 different species, and 33 from black rats captured around the monkey houses) and 7 water samples were collected in the Zoological Garden, and were examined for the prevalence of Y. enterocolitica serovar O:8. Serovar O:8 was isolated from 21 of 65 monkeys (32.3%) and 5 of 33 (15.2%) black rats (Rattus rattus). Furthermore, we examined the 30 isolates using molecular typing methods, pulsed field gel electrophoresis (PFGE), ribotyping using the RiboPrinter system, and restriction endonuclease analysis of virulence plasmid DNA (REAP), and compared the isolates in this outbreak with Japanese O:8 isolates previously identified. Genotyping showed that almost all the isolates were identical, and the genotype of the isolates was highly similar to that from wild rodents captured in Niigata Prefecture. This is the first report of fatal cases of Y. enterocolitica serovar O:8 infection in monkeys anywhere in the world.
Sequence Analysis of the Hypervariable Regions of the 56 kDa Immunodominant Protein Genes of Orientia tsutsugamushi Strains in Malaysia
Authors: Tay Sun Tee , Rohani Yasin Mohd , Ho Tze Ming , Shamala Devi 
Published: November 2013

AbstractThe DNA sequences encompassing two hypervariable regions, VD II and III of the 56 kDa immunodominant protein gene of 21 Malaysian strains of Orientia tsutsugamushi were determined. Two strains demonstrated a 100% DNA homology with the Gilliam prototype strain, and one with TH1817 strain and TA678 strain respectively. High percentages of DNA similarity (95–99%) were observed with Karp (4 strains), Gilliam (2 strains), TH1817 (4 strains), TC586 (3 strains) and TA763 (1 strain). The remaining strains demonstrated the highest DNA similarity with TA763 (1 strain, 89%), TA678 (1 strain, 86%) and TA686 (1 strain, 87%). Our study provides additional evidence on the existence and the genetic heterogeneity of TA strains of the Southeast Asia and their closely related strains in Malaysia.
Mutations in the gyrA and parC Genes in Ciprofloxacin‐Resistant Clinical Isolates of Acinetobacter baumannii in Korea
Authors: Lee Jeom Kyu , Lee Yeong Seon , Park Yong Keun , Kim Bong Su 
Published: November 2013

AbstractMutation with Ser‐83→Leu in gyrA gene was associated with the principal mutation for ciprofloxacin resistance in clinical isolates of Acinetobacter baumannii. Double mutation, Ser‐83→Leu in gyrA gene and Ser‐80→Leu in parC gene, was the most frequently detected among ciprofloxacin‐resistant isolates. A novel mutation with Ser‐80→Trp in parC gene, in addition to mutation in gyrA gene, was associated with a high‐level ciprofloxacin resistance. These results suggested that the presence of an additional mutation in the parC gene contributed to a higher‐level of ciprofloxacin resistance than a single mutation in the gyrA gene (geometric mean MICs of ciprofloxacin, 44.1 versus 16 μg/ml, P<0.05).
Molecular Subtyping for Escherichia coli O157:H7 Isolated in Taiwan
Authors: Tsai Tsung‐Yu , Luo Wei‐Chen , Wu Fang‐Tzy , Pan Tzu‐Ming 
Published: November 2013

AbstractEnterohaemorrhagic Escherichia coli O157:H7 is an important pathogen these days. Outbreaks of its infection have been reported all over the world, in Australia, Canada, Japan, the United States, south Africa, and various countries in Europe. In the summer of 2001, the first clinical infection by E. coli O157:H7 was identified in Taiwan. In this study, the standard procedures for molecular subtyping were applied to several strains collected in Taiwan as well as from elsewhere. The two molecular subtyping methods we used are pulsed‐field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP). The isolates from the U.S.A., Canada, Japan, and Taiwan each showed a unique molecular fingerprinting pattern. The environmental strains isolated in Taiwan showed closer relationships with each other, and their similarity was in the range of 75–85%. The first clinical strain isolated in Taiwan in 2001 was similar to the strains from North America but not closely related to the Taiwanese environmental strains. Our surveys showed that some local E. coli O157:H7 strains did exist in Taiwan, but there had been only one official case report of the infection by local E. coli O157:H7. The eating habits of the people and the geographic distribution of the pathogen are considered crucial risk factors in Taiwan. The establishment of a database of our own and joining the global network database are important tasks if we want to control such agricultural and food‐borne pathogens, and reduce the number of victims and amount sufferings, as well as the economic losses due to the infection.
Vaccine Efficacy of Recombinant GAPDH of Edwardsiella tarda against Edwardsiellosis
Authors: Liu Ying , Oshima Syun‐ichirou , Kurohara Kenrou , Ohnishi Kouhei , Kawai Kenji 
Published: November 2013

AbstractThirty‐seven kilodalton glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) of Edwardsiella tarda was suggested to be an effective vaccine candidate against E. tarda infection in previous research. For developing a vaccine, obtaining GAPDH in large quantities is necessary. In this study, we determined the complete nucleotide sequence of the gene that encodes GAPDH of E. tarda, and overexpressed the GAPDH of E. tarda by using the Escherichia coli expression system. We immunized Japanese flounder with recombinant GAPDH (rGAPDH) and evaluated its vaccine efficacy. Our results showed that rGAPDH effectively protected Japanese flounder from experimental E. tarda infection, and will contribute to the development of a vaccine against E. tarda.
Detection and Identification of Human Papillomavirus Types Isolated from Korean Patients with Flat Warts
Authors: Yoo Hoon , Won Sung‐Sik , Choi Hyun‐Chul , Yoon Taek‐Joon , Ye Sang‐Kyu , Park Tae‐Kyu , Lee Kwang‐Ho 
Published: November 2013

AbstractFlat warts, also called verruca planna (VP) or juvenile warts, are benign epithelial proliferations of the skin caused by infection with human papillomaviruses (HPV). Several HPV types are known to be associated with flat warts, and particularly HPV type 3 and 10 have been most frequently reported in other countries. In this study, for the detection and typing of human papillomavirus isolated from Korean patients with flat warts, polymerase chain reaction (PCR) and restriction endonuclease digestion were carried out with a set of restriction endonucleases, using the cloned HPV DNA and DNA from clinical specimens. A unique digestion pattern for HPV type 3 and 10, a form of miniature fingerprinting, enabled us to identify HPV type from the amplified fragments. A total of thirty clinical samples, as either frozen tissue or paraffin‐embedded tissue, were investigated to verify the type. All the clinical samples except one were confirmed to be type 3, one of the most frequently observed types in flat warts, and one sample was neither type 3 nor type 10. Further investigation of the unidentified sample by DNA sequencing and sequence alignment with other known HPV types revealed that the sample was a variant of HPV type 94, one of the EV‐related HPVs, with the closest evolutionary distance to the HPV type 10 among the known flat wart‐associated HPV types.
The Recombinant Immunogen with High‐Density Epitopes of ELDKWA and ELDEWA Induced Antibodies Recognizing Both Epitopes on HIV‐1 gp41
Authors: Wang Zuguang , Liu Zuqiang , Cheng Xiwen , Chen Ying‐Hua 
Published: November 2013

AbstractThe high mutation rate of HIV‐1 (human immunodeficiency virus‐1) is a major obstacle to developing an effective vaccine. The mutation of ELDKWA‐ (aa669–674) to ELDEWA‐epitope on HIV‐1 gp41 caused the immune escape from neutralization by potent anti‐HIV‐1 human monoclonal antibody (mAb) 2F5. In this study, we suggested and evaluated a multi‐epitope vaccine as a new strategy to develop HIV‐1 vaccines. A glutathione S‐transferase (GST) fusion protein (GST‐K8E8) containing 8 copies of ELDKWA‐ and mutated ELDEWA‐epitopes was constructed and used to immunize mice or rabbits. Analysis of the antisera (rAS3) induced by GST‐K8E8 suggested that multi‐epitope vaccine immunogen could raise antibodies in mice and rabbits against either the original ELDKWA‐epitope or the mutated ELDEWA‐epitope that resulted in immune escape. Briefly, ELDKWA‐epitope‐specific antibodies, directly purified from rAS3 by ELDKWA‐epitope‐peptide affinity chromatography, recognized either original gp41 protein (ELDKWA, rgp41K) or mutated gp41 (ELDEWA, rgp41E) in immunoblotting assay; in contrast, the existing ELDKWA‐epitope antibodies recognized only rgp41K but not rgp41E, which were purified by ELDKWA‐epitope‐peptide affinity chromatography from rAS3 that were firstly completely pre‐absorbed by ELDEWA‐epitope‐peptide affinity beads. And the same results were also observed when detecting the ELDEWA‐epitope‐specific antibodies in rAS3 by a means similar to the above. All the data presented here demonstrated that a high density multi‐epitope vaccine could be an interesting strategy against HIV‐1 mutation.
Rapid PrPSc Detection in Lymphoid Tissue and Application to Scrapie Surveillance of Fallen Stock in Japan: Variable PrPSc Accumulation in Palatal Tonsil in Natural Scrapie
Authors: Shimada Kimi , Hayashi Hiroko K. , Ookubo Yuka , Iwamaru Yoshifumi , Imamura Morikazu , Takata Masuhiro , Schmerr Mary Jo , Shinagawa Morikazu , Yokoyama Takashi 
Published: November 2013

AbstractRapid western blot (WB) procedure for an abnormal isoform of prion protein (PrPSc) detection in lymphoid tissues was established and has been applied to the surveillance of fallen stock. In this program, brain and palatal tonsil were examined by WB and three cases of sheep scrapie were detected. While one clinically scrapie‐infected sheep harbored PrPSc in the brain and palatal tonsil, the two sheep in the pre‐clinical stage harbored PrPSc in the brain, but not in the palatal tonsil. This study shows that PrPSc accumulation in palatal tonsil is variable in natural scrapie, even among genetically susceptible sheep.
Further Studies on the Mechanism of Rabies Virus Neutralization by a Viral Glycoprotein‐Specific Monoclonal Antibody, #1‐46‐12
Authors: Irie Takashi , Kawai Akihiko 
Published: November 2013

AbstractWe previously reported that a conformational epitope‐specific monoclonal antibody (mAb; #1‐46‐12) neutralized the rabies virus by binding only a small number (less than 20) of the antibody molecules per virion, while a linear epitope‐specific mAb (#7‐1‐9) required more than 250 IgG molecules for the neutralization. We also isolated both the epitope‐negative (R‐31) and ‐positive (R‐61) escape mutants that resisted mAb #1‐46‐12. Co‐infection studies with wild type (wt) and R‐61 mutant have shown that although the infectivity of R‐61 mutant was not affected by the binding of about 300 IgG molecules per virion, incorporation of a small number of wt G protein into the R‐61 virion resulted in dramatic loss of the resistance. In this study, we further investigated properties of the mutant G proteins. The R‐61 G protein lost reactivity to the mAb when solubilized, even keeping a trimer form, suggesting that membrane‐anchorage is essential for the maintenance of its epitope‐positive conformation. On the other hand, incorporation of wt G proteins into the R‐31 virions did not affect their resistance to the mAb very much. Although we have not so far found the presumed conformational changes induced by the mAb‐binding, we think that these results are not inconsistent with our previously proposed novel model (referred to as a domino effect model) for the virus neutralization by mAb #1‐46‐12 other than a classical spike‐blocking model, which implicates successive spreading of the postulated antibody‐induced conformational changes of G protein to the neighboring spikes until abolishing the host cell‐binding ability of the virion.
A Novel B‐2 Suppressor Cell Regulating Susceptibility/Resistance of Mice to Toxoplasma gondii Infection
Authors: Mun Hye‐Seong , Aosai Fumie , Fang Hao , Piao Lian‐Xun , Winn Tin , Norose Kazumi , Yano Akihiko 
Published: November 2013

AbstractIrradiation treatment enhanced resistance of C57BL/6, but not BALB/c against Toxoplasma gondii infection. Six Gy‐irradiated (IR) C57BL/6 recipients of B‐2 cells from T. gondii‐infected C57BL/6 died after infection. B‐2 suppressor cells from infected C57BL/6 enhanced production of IL‐4 and IL‐10 in peritoneal exudate cells (PECs), and down‐regulated NO release in peritoneal macrophages after infection. On the other hand, B‐2 suppressor cells were not detected in a strain, BALB/c, resistant against infection. These data indicated that irradiation‐sensitive B‐2 cells regulated susceptibility/resistance in mice against T. gondii infection.
Rapid Detection and Differentiation of the Major Mycoplasma Contaminants in Cell Cultures Using Real‐Time PCR with SYBR Green I and Melting Curve Analysis
Authors: Harasawa Ryô , Mizusawa Hiroshi , Fujii Masashi , Yamamoto Junko , Mukai Hiroyuki , Uemori Takashi , Asada Kiyozo , Kato Ikunoshin 
Published: November 2013

AbstractA quantitative real‐time polymerase chain reaction (PCR) procedure followed by melting curve analysis, using the green fluorescence dye SYBR Green I, was developed for rapid detection and differentiation of mycoplasma contaminants in cell cultures. This method showed that the detection of the target sequence was linear over a range from 104 to 10 colony‐forming units (CFU) of the mycoplasma cells. Analysis of the melting temperature of the PCR products allowed differentiation of the major mycoplasma contaminants. These results demonstrate that the protocol described in the present study can decrease the time to obtain reproducible results by simultaneous detection and differentiation of the Mycoplasma species contaminating cell cultures.
Identification of an HLA‐A24‐Restricted OY‐TES‐1 Epitope Recognized by Cytotoxic T‐Cells
Authors: Okumura Hideo , Noguchi Yuji , Uenaka Akiko , Aji Toshiki , Ono Toshiro , Nakagawa Kazuhiko , Aoe Motoi , Shimizu Nobuyoshi , Nakayama Eiichi 
Published: November 2013

AbstractOY‐TES‐1 was identified as a human homologue of the mouse, guinea pig, and pig proacrosin binding protein sp32 precursor. Differential expression levels of OY‐TES‐1 mRNA between testis and other normal tissues, and its expression in cancers indicated that OY‐TES‐1 would be classified as a cancer/testis antigen and considered to be a candidate of target antigen for cancer immunotherapy. In this study, we showed identification of HLA‐A24‐binding OY‐TES‐1 peptide, TES401–409 (KTPFVSPLL) recognized by CD8 T‐cells. Purified CD8 T‐cells from healthy donors stimulated in vitro with the peptide‐pulsed autologous DC and PBMC produced IFN7 in response to the peptide‐pulsed PBMC and showed cytotoxicity against the peptide‐pulsed autologous EBV‐B specifically. Furthermore, cytotoxicity was also observed against an OY‐TES‐1 mRNA‐expressing tumor line, LK79. The retention time of the fraction in HPLC of the acid eluate from LK79 cells that showed positive sensitization against autologous EBV‐B cells in recognition by CD8 CTL was the same as that of the fraction of the TES401–409 peptide itself, suggesting that the TES401–409 was a naturally processed peptide on LK79.
Characterization of M Gene‐Deficient Rabies Virus with Advantages of Effective Immunization and Safety as a Vaccine Strain
Authors: Ito Naoto , Sugiyama Makoto , Yamada Kentaro , Shimizu Kenta , Takayama‐Ito Mutsuyo , Hosokawa Junji , Minamoto Nobuyuki 
Published: November 2013

AbstractMatrix (M) protein of rabies virus is known to play an important role in assembly and budding of the progeny virus. We generated an M gene‐deficient rabies virus, RC‐HLAM, using a reverse genetics system of rabies virus RC‐HL strain to develop a novel type of vaccine. RC‐HLAM infection was confined within a single cell in mouse neuroblastoma cells. This deficient virus failed to generate the progeny virus in the cells. In contrast, RC‐HLAM propagated in BHK cells inductively expressing M protein. Suckling and adult mice inoculated intracerebrally with the parental RC‐HL strain showed lethal infection and transient body weight loss, respectively, whereas both suckling and adult mice inoculated with RC‐HLAM showed no symptoms. The neutralizing antibody against rabies virus was successfully induced by intramuscular immunization with 105 focus‐forming units of RC‐HLAM but not UV‐inactivated RC‐HL. Intranasal immunization with RC‐HLAM resulted in almost the same antibody titer to rabies virus as that in the case of immunization with live RC‐HL strain. These findings indicate that RC‐HLAM is a candidate for a novel rabies vaccine that is safer and more effective than are current vaccines.
Contribution of Anaphylatoxins to Allergic Inflammation in Human Lungs
Authors: Abe Masayoshi , Hama Hiroshi , Shirakusa Takayuki , Iwasaki Akinori , Ono Nobuhumi , Kimura Nobuhiro , Hugli Tony E. , Okada Noriko , Katsuragi Takeshi , Okada Hidechika 
Published: November 2013

AbstractThe contribution of complement activation to allergic asthma remains controversial. In order to elucidate the role played by the complement split products, anaphylatoxins C3a and C5a, we evaluated their effects on production of cysteinyl‐leukotrienes (cysLTs) by human lung fragments following an anaphylactic reaction. The lung tissues obtained from two patients with lung cancer showed C5aR‐, C5L2R‐, and C3aR‐mRNA expression. When the chopped lung fragments passively sensitized with human IgE were incubated with anti‐human IgE antibody, a significant amount of cysLTs was generated in comparison with the control (without anti‐IgE antibody). The co‐addition of human C5a at doses of 0.1 to 10 ng/ml to the anti‐IgE antibody potentiated cysLT production. The response was bell‐shaped in distribution, significant, and peaked at a C5a concentration of 1 ng/ml. The co‐addition of human C3a up to 1,000 ng/ml seemed to increase cysLT production, but not to any significant extent. A novel C5a receptor complementary peptide, acetylated peptide A, dose‐dependently inhibited cysLT production by the human lung fragments following the anaphylactic reaction in the presence of 1 ng/ml C5a. However, this peptide did not inhibit cysLT production in the presence of 100 ng/ml C3a. It is suggested that the anaphylatoxin C5a potentiates cysLT production in human lung tissues and contributes to allergic inflammation in disorders such as asthma, thus acetylated peptide A may be useful for suppressing allergic inflammation in the lungs.
Evaluation of the Lon‐Deficient Salmonella Strain as an Oral Vaccine Candidate
Authors: Kodama Chie , Eguchi Masahiro , Sekiya Yukie , Yamamoto Tomoko , Kikuchi Yuji , Matsui Hidenori 
Published: November 2013

AbstractWe evaluated the efficacy of CS2022 (the Lon protease‐deficient mutant strain of Salmonella enterica serovar Typhimurium) as a candidate live oral vaccine strain against subsequent oral challenge with a virulent strain administered to BALB/c and C57BL/6 mice. CS2022 persistently resided in the spleen, mesenteric lymph nodes, Peyer's patches, and cecum of both strains of mice after a single oral inoculation with 1 × 108 colony‐forming units. Finally, CS2022 almost disappeared from each tissue sample by week 12 in BALB/c mice, whereas CS2022 still resided in each tissue type at week 12 after inoculation of C57BL/6 mice. A significant increase in the serovar Typhimurium lipopolysaccharide‐specific secretory immunoglobulin A (s‐IgA), as measured for one of the mucosal immune responses, was detected in bile and intestinal samples of both strains of immunized mice at week 4 after immunization. In addition, the expression of gamma interferon mRNA in the spleens of both strains of immunized mice, especially those of C57BL/6 mice, was significantly increased at week 4 after immunization and was boosted during the following 5 days after the challenge was administered to the mice. Furthermore, peritoneal macrophages isolated from immunized mice at week 4 after immunization exhibited an increase in intracellular killing activity against both virulent and avirulent Salmonella. The present results suggested that salmonellae‐specific s‐IgA on the mucosal surfaces induced by immunization with CS2022 generally prevented mice from succumbing to an oral challenge with a virulent strain. Simultaneously, CS2022 promoted the protective immunity associated with macrophages in both strains of mice.