bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of the Protease Activity of Influenza Virus RNA Polymerase PA Subunit by Viral Matrix Protein
Authors: Hara Koyu , Shiota Mayumi , Kido Hiroshi , Watanabe Ken , Nagata Kyosuke , Toyoda Tetsuya 
Published: November 2013

AbstractInfluenza virus PA is a subunit of RNA‐dependent RNA polymerase. We demonstrated that PA has a unique chymotrypsin‐like serine protease activity with Ser624 as an active site. To obtain further insight into the role of the protease activity of PA in viral proliferation, we examined the interaction between PA and matrix protein (M1). Both M1 purified from virion and hexa‐histidine‐tagged M1 expressed in Escherichia coli bound to PA. Hexa‐histidine‐tagged M1 pulled down PA. The interaction of PA with M1 was sensitive to ionic strength, suggesting that the interaction is formed by electrostatic force. Using Suc‐Leu‐Leu‐Val‐Tyr‐MCA, a specific substrate for PA protease, M1 was demonstrated to inhibit the amidolytic activity of PA, whereas M1 did not inhibit that of chymotrypsin or trypsin at all. These results suggest that M1 binds to and inhibits the amidolytic activity of PA.
Prevalence of Inherited Myeloperoxidase Deficiency in Japan
Authors: Nunoi Hiroyuki , Kohi Fumikazu , Kajiwara Hideki , Suzuki Kazuo 
Published: November 2013

AbstractThe microbicidal activity of the myeloperoxidase (MPO)‐hydrogen peroxide‐halide system has been implicated as the most efficient, oxygen‐dependent antimicrobial component of neutrophil host defense. Unexpectedly, individuals with MPO deficiency suffer few clinical consequences. In order to understand better the clinical impact of MPO deficiency, we surveyed several clinical hematology laboratories in Japan to assess the prevalence of MPO deficiency in the general population. MPO activity was determined by flow cytometry using the Technicon H series of automated systems. We identified 26 cases of complete MPO deficiency, prevalence 1 in 57,135, and 129 cases of partial deficiency, prevalence 1 in 17,501. The distribution of complete and partial deficiencies differed among the laboratories studied.
Cloning and Expression of Bartonella henselae sucB Gene Encoding an Immunogenic Dihydrolipoamide Succinyltransferase Homologous Protein
Authors: Kabeya Hidenori , Maruyama Soichi , Hirano Kouji , Mikami Takeshi 
Published: November 2013

AbstractImmunoscreening of a ZAP genomic library of Bartonella henselae strain Houston‐1 expressed in Escherichia coli resulted in the isolation of a clone containing 3.5 kb BamHI genomic DNA fragment. This 3.5 kb DNA fragment was found to contain a sequence of a gene encoding a protein with significant homology to the dihydrolipoamide succinyltransferase of Brucella melitensis (sucB). Subsequent cloning and DNA sequence analysis revealed that the deduced amino acid sequence from the cloned gene showed 66.5% identity to SucB protein of B. melitensis, and 43.4 and 47.2% identities to those of Coxiella burnetii and E. coli, respectively. The gene was expressed as a His‐Nus A‐tagged fusion protein. The recombinant SucB protein (rSucB) was shown to be an immunoreactive protein of about 115 kDa by Western blot analysis with sera from B. henselae‐immunized mice. Therefore the rSucB may be a candidate antigen for a specific serological diagnosis of B. henselae infection.
Phylogenetic Characterization of Orientia tsutsugamushi Isolated in Taiwan According to the Sequence Homologies of 56‐kDa Type‐Specific Antigen Genes
Authors: Qiang Yu , Tamura Akira , Urakami Hiroshi , Makisaka Yukie , Koyama Shoichiro , Fukuhara Masahiro , Kadosaka Teruki 
Published: November 2013

AbstractFourteen strains of Orientia tsutsugamushi isolated in Taiwan were characterized by sequencing 56‐kDa type‐specific antigen genes and patterns of restriction fragment length polymorphism (RFLP) predicted by a computer program. The strains showed high varieties in sequence homologies and were classified to 10 types by predicted patterns of RFLP. Furthermore, all Taiwan strains were not identical in typing with strains analyzed previously. These results suggest that there are various types of O. tsutsugamushi in Taiwan that are different from those distributed in other countries.
The Inhibitory Effect of Essential Oils on Herpes Simplex Virus Type‐1 Replication In Vitro
Authors: Minami Masato , Kita Masakazu , Nakaya Takaaki , Yamamoto Toshiro , Kuriyama Hiroko , Imanishi Jiro 
Published: November 2013

AbstractThe antiviral effect of 12 essential oils on herpes simplex virus type‐1 (HSV‐1) replication was examined in vitro. The replication ability of HSV‐1 was suppressed by incubation of HSV‐1 with 1% essential oils at 4 C for 24 hr. Especially, lemongrass completely inhibited the viral replication even at a concentration of 0.1%, and its antiviral activity was dependent on the concentrations of the essential oil. When Vero cells were treated with the essential oil before or after viral adsorption, no antiviral activity was found, which suggests that the antiviral activity of essential oils including lemongrass may be due to the direct interaction with virions.
Sensitive and Rapid Quantitative Detection of Anthrax Spores Isolated from Soil Samples by Real‐Time PCR
Authors: Ryu Chunsun , Lee Kyunghee , Yoo Cheonkwon , Seong Won Keun , Oh Hee‐Bok 
Published: November 2013

AbstractQuantitative analysis of anthrax spores from environmental samples is essential for accurate detection and risk assessment since Bacillus anthracis spores have been shown to be one of the most effective biological weapons. Using TaqMan real‐time PCR, specific primers and probes were designed for the identification of pathogenic B. anthracis strains from pag gene and cap gene on two plasmids, pXO1 and pXO2, as well as a sap gene encoded on the S‐layer. To select the appropriate lysis method of anthrax spore from environmental samples, several heat treatments and germination methods were evaluated with multiplex‐PCR. Among them, heat treatment of samples suspended with sucrose plus non‐ionic detergent was considered an effective spore disruption method because it detected up to 105 spores/g soil by multiplex‐PCR. Serial dilutions of B. anthracis DNA and spore were detected up to a level of 0.1 ng/μl and 10 spores/ml, respectively, at the correlation coefficient of 0.99 by real‐time PCR. Quantitative analysis of anthrax spore could be obtained from the comparison between CT value and serial dilutions of soil sample at the correlation coefficient of 0.99. Additionally, spores added to soil samples were detected up to 104 spores/g soil within 3 hr by real‐time PCR. As a consequence, we established a rapid and accurate detection system for environmental anthrax spores using real‐time PCR, avoiding time and labor‐consuming preparation steps such as enrichment culturing and DNA preparation.
Differential Expression and Signaling of IFN‐γ in the Conjunctiva between Lewis and Brown Norway Rats
Authors: Fukushima Atsuki , Ozaki Akemi , Fukata Kazuyo , Ueno Hisayuki 
Published: November 2013

AbstractGenetic background determines the histological features of experimental immune‐mediated blepharoconjunctivitis (EC) in rats, which is a model for human allergic conjunctivitis (AC). A great number of lymphocytes predominate in EC of Lewis rats, while less lymphocytes and more eosinophils are present in that of Brown Norway (BN) rats. Although this difference could be attributed to their systemic Th1/Th2 dominancy, it remains unclear whether some regulatory mechanisms may exist in the inflammatory site in the conjunctiva. Here, we aim to investigate this hypothesis by comparing the expression levels of inflammatory mediators in the conjunctiva between the two strains. EC was induced in Lewis and BN rats by transfer of ovalbumin (OVA)‐specific CD4+ T‐cell lines followed by eye drops of OVA as antigen challenge, and then was clinically and histologically evaluated. Reverse‐transcription (RT)‐PCR was performed to compare the expressions of cytokines and cytokine receptors (Rs) in conjunctivas of both strains of rats either with or without EC. To confirm the biological significance of interferon (IFN)‐γ R expression, phosphorylation of signal transducers and activators of transcription (STAT)‐1 was examined in the conjunctivas, followed by subconjunctival injection of IFN‐γ. BN T cells contained interleukin (IL)‐4 and IFN‐γ, while Lewis T cells expressed no IL‐4. Transfer of those cells induced more severe EC in Lewis rats. RT‐PCR using naive conjunctivas detected more IL‐4, IFN‐γ, and IFN‐γR β‐chain RNA expression in BN rats. After the EC induction, BN rats expressed significantly higher amounts of IFN‐γR β‐chain, and upregulation of interferon regulatory factor (IRF)‐1 was observed. Phosphorylation of STAT‐1 was more remarkable in BN rats. The findings demonstrate differential expression of IFN‐γR and signaling through IFN‐γ in the conjunctiva between the two strains. This may be due to differences in histopathological character between the two strains.
Distribution of the usp Gene in Uropathogenic Escherichia coli Isolated from Companion Animals and Correlation with Serotypes and Size‐Variations of the Pathogenicity Island
Authors: Kurazono Hisao , Nakano Masayuki , Yamamoto Shingo , Ogawa Osamu , Yuri Kazuyo , Nakata Katsuhisa , Kimura Miyuki , Makino Sou‐ichi , Nair G. Balakrish 
Published: November 2013

AbstractUncomplicated urinary tract infection (UTI) caused by uropathogenic Escherichia coli (UPEC) is a serious problem not only among humans but also in companion animals such as dogs and cats. The uropathogenic specific protein gene (usp) is preferentially distributed in UPEC isolates from dogs and cats compared with the distribution of usp in E. coli strains from feces of healthy dogs and cats and this pattern of distribution resembles that observed in human UPEC strains. The UPEC strains from companion animals share common O serotypes like O1, O2, O4, O6, O16, O18, O22, O25 and O75 as those reported for human UPEC. The size variation of the pathogenicity island that includes usp in UPEC from dogs and cats was almost similar to those seen in human UPEC. We propose that dogs and cats are the alternative reservoirs for UPEC strains that are associated with human UTI.
Phylogenetic Analysis of Spotted Fever Group Rickettsiae Based on gltA, 17‐kDa, and rOmpA Genes Amplified by Nested PCR from Ticks in Japan
Authors: Ishikura Mitsuhiro , Ando Shuji , Shinagawa Yasuhiro , Matsuura Kumiko , Hasegawa Sumiyo , Nakayama Takashi , Fujita Hiromi , Watanabe Mamoru 
Published: November 2013

AbstractIn order to understand the natural situation of rickettsiae in the ticks in Japan, the rickettsial genes, gltA gene, rOmpA gene, and 17‐kDa gene, were amplified from the ticks by nested PCR. The prevalences of rickettsial gltA genes among Haemaphysalis formosensis, H. longicornis, H. megaspinosa, Ixodes ovatus, H. flava, H. kitaokai, and I. persulcatus were 62, 57, 24, 24, 19, 13, and 10%, respectively; 26% (186/722) being the average. The gltA genes amplified from the ticks were classified into 9 genotypes (I to IX) by the difference in nucleotide sequences. Genotype I was detected from 7 species of ticks. Genotype II mainly was detected from H. longicornis and H. formosensis. Genotypes III and VII mainly were detected from H. flava and I. ovatus. The polarization in the distribution of genotypes among regions where the ticks were collected was not clear. Based on the phylogenetic analysis of the three genes presented here, genotypes I, III, and IV (detected from H. formosensis, H. hystricia, and I. ovatus) are genetically close with each other, but rickettsiae of the same property still have not been isolated from ticks anywhere in the world. These genotypes should be considered as new species among SFG rickettsiae. Genotype II was identical with strain FUJ‐98, genetically close to R. japonica which has been isolated from ticks in China. Genotype V was identical with R. felis and strain California 2 isolated from the cat flea. This is the first report on the detection of R. felis from ticks. Genotype VI detected from Ixodes sp. did not seem to belong to genus Rickettsia. Based on the previous antigenic data and the phylogenetic analysis presented here, Genotype VII should be considered a variant of R. helvetica and genotype VIII detected from I. ovatus and I. persulcatus were identical with R. helvetica. Genotype IX detected from I. nipponensis was genetically close to the strains IRS3, IRS4, and IrR/Munich isolated from I. ricinus in Slovakia and German.
Mycobacterial Infection in MyD88‐Deficient Mice
Authors: Sugawara Isamu , Yamada Hiroyuki , Mizuno Satoru , Takeda Kiyoshi , Akira Shizuo 
Published: November 2013

AbstractMyD88 is an adaptor protein that plays a major role in TLR/IL‐1 receptor family signaling. To understand the role of MyD88 in the development of murine tuberculosis in vivo, MyD88 knockout (KO) mice aerially were infected with Mycobacterium tuberculosis. Infected MyD88 mice were not highly susceptible to M. tuberculosis infection, but they developed granulomatous pulmonary lesions with neutrophil infiltration which were larger than those in wild‐type (WT) mice (P<0.01). The pulmonary tissue levels of mRNA for iNOS and IL‐18 were slightly lower, but levels of mRNA for IL‐1β, IL‐2, IL‐4, IL‐6, IL‐10, IFN‐γ, and TGF‐β were higher in MyD88 KO mice. IFN‐γ, TNF‐α, IL‐1β, and IL‐12 also were high in the sera of MyD88 KO mice. There were no statistically significant differences in the expression of TNF‐α, IL‐12, and ICAM‐1 mRNA between MyD88 KO and WT mice. Thus, MyD88 deficiency did not influence the development of murine tuberculosis. NF‐κB activity was similar in the alveolar macrophages from the lung tissues of MyD88 KO and WT mice. Also, there may be a TLR2‐specific, MyD88‐independent IL‐1 receptor/TLR‐mediated pathway to activate NF‐κB in the host defense against mycobacterial infection.
Molecular Epidemiology of Bordetella pertussis Isolated in Taiwan, 1992–1997
Authors: Lee Yeong‐Sheng , Yang Chiou‐Ying , Lu Cheng‐Hsiung , Tseng Yi‐Hsiung 
Published: November 2013

AbstractIn Taiwan, the number of pertussis cases including various types of infection has been increasing in recent years, especially in 1997. Since 71% of the reported cases concentrated in the densely populated Taipei metropolitan area, concerns have been raised that a highly contagious strain of Bordetella pertussis might have appeared in Taipei. In this study, 114 strains of B. pertussis including those isolated in 1992–1996 (n = 53) and 1997 (n=61) were subjected to pulsed‐field gel electrophoresis (PFGE) of the XbaI digests from their chromosomes. Based on the band patterns, they were divided into 21 subtypes, P1 to P21. The strains isolated in 1997 consist of 17 subtypes including 9 new subtypes which did not appear in the previous years, indicating that the outbreaks in 1997 were not caused by a sole specific virulent strain. Dendrogram analysis indicated that the 21 subtypes can be grouped into five clusters, with the first four subtypes possessing 60 to 95% relatedness to one another, whereas relatedness between cluster 5 (containing P21 only) and the other clusters is less than 50%. Notably, all the subtypes except P12 and P21 appeared at least once in Taipei and the majority of the strains (54%) belong to two clusters, 3 and 4. These results suggest that highly dense population may facilitate spread and accelerate genetic divergence of this pathogen. This is the first report on pertussis molecular epidemiology in Taiwan.
Effects of High Amylose Maize Starch and Clostridium butyricum on Metabolism in Colonic Microbiota and Formation of Azoxymethane‐Induced Aberrant Crypt Foci in the Rat Colon
Authors: Nakanishi Shuusuke , Kataoka Keiko , Kuwahara Tomomi , Ohnishi Yoshinari 
Published: November 2013

AbstractHigh amylose maize starch (HAS) is not digested in the small intestine and most of it reaches the large intestine. In the large intestine, HAS is fermented by intestinal bacteria, resulting in production of short‐chain fatty acids (SCFA), particularly butyrate. Clostridium butyricum can utilize HAS and produce butyrate and acetate. It has been proposed that butyrate inhibits carcinogenesis in the colon. In this study, we examined the inhibitory effects of HAS and C. butyricum strain MIYAIRI588 (CBM588) on azoxymethane‐induced aberrant crypt foci (ACF) formation in rats. In the group of rats administered only CBM588 spores, the concentration of butyrate in the cecum increased, but there was no decrease in the number of ACF. In the group of rats fed an HAS diet, a decrease in the number of ACF was observed, and in the group of rats administered HAS and CBM588, the number of ACF decreased significantly. In these two groups, the concentrations of acetate and propionate in intestinal contents significantly increased, but the concentration of butyrate did not change. It was found that the β‐glucuronidase activity level of colonic contents decreased significantly in the two groups of rats fed HAS. This study showed that HAS and CBM588 changed the metabolism of colonic microbiota and decreased the level of β‐glucuronidase activity, phenomena that may play a role in the inhibition of ACF formation in the rat colon.
Serum Thymic Factor Prevents LPS‐Induced Pancreatic Cell Damage in Mice via Up‐Regulation of Bcl‐2 Expression in Pancreas
Authors: Saitoh Noriko , Awaya Akira , Sakudo Akikazu , SungWook Seo , Saeki Keiichi , Matsumoto Yoshitsugu , Onodera Takashi 
Published: November 2013

AbstractThe protective effect of synthetic serum thymic factor (FTS) nonapeptide on lipopolysaccharide (LPS)‐induced pancreatic cell damage in 10‐week‐old BALB/c male mice was investigated. Mice were divided into three groups. Group I was treated with LPS (10 μg/head; i.p.) (LPS‐treated mice). Group II was administered with FTS (50 μg/head; i.p.) 24 hr before treatment with LPS and complemented immediately before LPS injection with FTS (50 μg/head; i.p.) (FTS‐administered mice). Group III was only treated with the same volume of saline (control mice). Treatment of LPS in vivo resulted in the destruction of pancreatic acinar cells. In those cells, many apoptotic cells were detected by immunohistochemistry using an anti‐single stranded DNA antibody. Immunohistochemistry and reverse transcription‐polymerase chain reaction (RT‐PCR) revealed that LPS treatment also caused low or a lack of insulin expression in pancreatic islets. In contrast, morphological change was not seen and apoptotic cell death was suppressed in pancreatic cells of FTS‐administered mice. Moreover, insulin expression was normal in those mice. FTS administration enhanced expression of Bcl‐2 mRNA levels in pancreatic tissues and IL‐6 mRNA levels in splenocytes significantly compared with those of LPS treatment at 3 hr after LPS injection. These findings suggest that FTS prevents LPS‐induced cell damage via enhancing Bcl‐2 expression in the pancreas and systemic IL‐6 production.
Evaluation of Three Different Forward Primers by Terminal Restriction Fragment Length Polymorphism Analysis for Determination of Fecal Bifidobacterium Spp. in Healthy Subjects
Authors: Hayashi Hidenori , Sakamoto Mitsuo , Benno Yoshimi 
Published: November 2013

AbstractThe 27F forward primer is frequently used in 16S rRNA gene libraries and T‐RFLP analysis. However, Bifidobacterium spp. were barely detected with this primer in human fecal samples. In this study, fecal microbiota were analyzed using the T‐RFLP method with three different forward primers (27F, 35F, and 529F) in conjunction with one reverse primer (1492R). T‐RFLP analysis of fecal microbiota using 35F and 529F detected higher proportions of the terminal restriction fragment (T‐RF) corresponding to Bifidobacterium spp. than that using 27F. 27F is in imperfect agreement while 35F and 529F are in good concordance with the 16S rRNA gene sequences of Bifidobacterium spp., and the latter primers allowed for the detection of T‐RFs of Bifidobacterium spp. in fecal samples from five healthy subjects. The T‐RFs presumed to be Bifidobacterium spp. were cloned and sequenced, and found to match the 16S rRNA gene sequences of Bifidobacterium spp. Among the five fecal samples, two samples with low frequencies of T‐RFs of Bifidobacterium spp. were detected using these forward primers. This probably reflects a low prevalence of Bifidobacterium spp. in these two samples. Our study emphasizes the importance of selecting a suitable forward primer for detection of Bifidobacterium spp.
Differential Susceptibility of Cells Expressing Allogeneic MHC or Viral Antigen to Killing by Antigen‐Specific CTL
Authors: Lee Koutetsu , Takenaka Hiroshi , Yoneda Yukio , Goto Toshiyuki , Sano Kouichi , Nakanishi Mahito , Eguchi Akiko , Okada Masashi , Tashiro Junko , Sakurai Kanji , Kubota Takahiro , Yoshida Ryotaro 
Published: November 2013

AbstractCD8+ cytotoxic T lymphocytes (CTLs) generated by immunization with allogeneic cells or viral infection are able to lyse allogeneic or virally infected in vitro cells (e.g., lymphoma and mastocytoma). In contrast, it is reported that CD8+ T cells are not essential for allograft rejection (e.g., heart and skin), and that clearance of influenza or the Sendai virus from virus‐infected respiratory epithelium is normal or only slightly delayed after a primary viral challenge of CD8‐knockout mice. To address this controversy, we generated H‐2d‐specific CD8+ CTLs by a mixed lymphocyte culture and examined the susceptibility of a panel of H‐2d cells to CTL lysis. KLN205 squamous cell carcinoma, Meth A fibrosarcoma, and BALB/c skin components were found to be resistant to CTL‐mediated lysis. This resistance did not appear to be related to a reduced expression of MHC class I molecules, and all these cells could block the recognition of H‐2d targets by CTLs in cold target inhibition assays. We extended our observation by persistently infecting the same panel of cell lines with defective‐interfering Sendai virus particles. The Meth A and KLN205 lines infected with a variant Sendai virus were resistant to lysis by Sendai virus‐specific CTLs. The Sendai virus‐infected Meth A and KLN205 lines were able to block the lysis of Sendai virus‐infected targets by CTLs in cold target inhibition assays. Taken together, these results suggest that not all in vivo tissues may be sensitive to CTL lysis.
Mechanisms of Action of Corilagin and Tellimagrandin I That Remarkably Potentiate the Activity of β‐Lactams against Methicillin‐Resistant Staphylococcus aureus
Authors: Shiota Sumiko , Shimizu Masato , Sugiyama Jun'ichi , Morita Yuji , Mizushima Tohru , Tsuchiya Tomofusa 
Published: November 2013

AbstractCorilagin and tellimagrandin I are polyphenols isolated from the extract of Arctostaphylos uvaursi and Rosa canina L. (rose red), respectively. We have reported that corilagin and tellimagrandin I remarkably reduced the minimum inhibitory concentration (MIC) of β‐lactams in methicillin‐resistant Staphylococcus aureus (MRSA). In this study, we investigated the effect of corilagin and tellimagrandin I on the penicillin binding protein 2′ (2a) (PBP2′ (PBP2a)) which mainly confers the resistance to β‐lactam antibiotics in MRSA. These compounds when added to the culture medium were found to decrease production of the PBP2′ (PBP2a) slightly. Using BOCILLIN FL, a fluorescent‐labeled benzyl penicillin, we found that PBP2′ (PBP2a) in MRSA cells that were grown in medium containing corilagin or tellimagrandin I almost completely lost the ability to bind BOCILLIN FL. The binding activity of PBP2 and PBP3 were also reduced to some extent by these compounds. These results indicate that inactivation of PBPs, especially of PBP2′ (PBP2a), by corilagin or tellimagrandin I is the major reason for the remarkable reduction in the resistance level of β‐lactams in MRSA. Corilagin or tellimagrandin I suppressed the activity of β‐lactamase to some extent.