bluuebunny/crossref_metadata_2025_split huggingface dataset:
Survival of Virulent Mycobacterium tuberculosis Involves Preventing Apoptosis Induced by Bcl‐2 Upregulation and Release Resulting from Necrosis in J774 Macrophages
Authors: Zhang Jianling , Jiang Ru , Takayama Hisao , Tanaka Yoshinori 
Published: November 2013

AbstractMacrophage apoptosis plays a role in mycobacterial infection. To define the mechanism by which virulent Mycobacterium tuberculosis escapes apoptosis and killing in macrophages, J774 macrophages were infected with virulent M. tuberculosis H37Rv and attenuated H37Ra strains. H37Rv induced less apoptosis than H37Ra, and caspase 3 was activated in H37Ra‐ and H37Rv‐infected macrophages. Intracellular H37Rv bacilli were released at a higher rate into the supernatant than were H37Ra by the sixth day of infection, and this was simultaneously accompanied by the increased necrosis of infected cells showing lactate dehydrogenase (LDH) release. Fas mRNA expression was downregulated and FasL was upregulated in H37Ra‐ and H37Rv‐infected macrophages, while Bcl‐2 was upregulated in H37Rv‐infected macrophages but downregulated in H37Ra‐infected macrophages as seen by real‐time PCR. These results indicate that M. tuberculosis H37Ra and H37Rv proliferate in macrophages by preventing them from inducing apoptosis during the early phase of infection, and that M. tuberculosis H37Rv‐infected macrophages are found to express Bcl‐2 mRNA, which leads to anti‐apoptotic activity, and that relatively distinct necrosis might occur during the later phase of infection.
Curli Fibers Are Required for Development of Biofilm Architecture in Escherichia coli K‐12 and Enhance Bacterial Adherence to Human Uroepithelial Cells
Authors: Kikuchi Tatsuya , Mizunoe Yoshimitsu , Takade Akemi , Naito Seiji , Yoshida Shin‐ichi 
Published: November 2013

AbstractSessile bacteria show phenotypical, biochemical, and morphological differences from their planktonic counterparts. Curli, extracellular structures important for biofilm formation, are only produced at temperatures below 30 C in Escherichia coli K‐12 strains. In this report, we show that E. coli K‐12 can produce curli at 37 C when grown as a biofilm community. The curli‐expressing strain formed more biofilms on polyurethane sheets than the curli‐deficient strain under growth temperatures of both 25 C and 37 C. Curli are required for the formation of a three‐dimensional mature biofilm, with characteristic water channels and pillars of bacteria. Observations by electron microscopy revealed the presence at the surfaces of the curli‐deficient mutant in biofilm of flagella and type I pili. A wild‐type curli‐expressing E. coli strain significantly adhered to several lines of human uroepithelial cells, more so than an isogenic curli‐deficient strain. The finding that curli are expressed at 37 C in biofilm and enhance bacterial adherence to mammalian host cells suggests an important role for curli in pathogenesis.
Immune Response to the Rhodococcus equi Infection in High and Low Antibody‐Producing Mice (Selection IV‐A)
Authors: Pedrini Sílvia C.B. , Acorci Michele J. , Pinto João G.G. , Silveira Liciana V.A. , Oliveira Silvio L. 
Published: November 2013

AbstractRhodococcus equi is a Gram‐positive, facultative intracellular bacterium which infects macrophages and causes rhodococcal pneumonia and enteritis in foals. Recently, this agent has been recognized as an opportunistic pathogen for immunocompromised humans. Several murine experimental models have been used to study R. equi infection. High (HIV‐A) and Low (LIV‐A) antibody (Ab)‐producers mice were obtained by bi‐directional genetic selections for their ability to produce antibodies against sheep and human erythrocytes (Selection IV‐A). These lines maintain their phenotypes of high and low responders also for other antigens than those of selection (multispecific effect). A higher macrophage activity in LIV‐A mice has been described for several intracellular infectious agents, which could be responsible for their intense macrophage antigens (Ag)‐handling and low Ab production. Due to these differences, LIV‐A mice were found to exhibit a better performance to trigger an effective immune response towards intracellular pathogens. The objective of this work was to characterize the immune response of Selection IV‐A against R. equi. HIV‐A and LIV‐A mice were infected with 2.0 ×106 CFU of ATCC 33701+R. equi by intravenous route. With regards to bacterial clearance and survival assays, LIV‐A mice were more resistant than HIV‐A mice to virulent R. equi. LIV‐A mice presented a higher hydrogen peroxide (H2O2) and nitric oxide (NO) endogenous production by splenic macrophages than HIV‐A mice. LIV‐A expressed the most intense cellular response, available by the Delayed‐Type Hypersensitivity (DTH) reaction, which activated macrophages and produced more H2O2 and NO. The three times higher specific antibodies titres in HIV‐A indicated that Selection IV‐A maintained the multispecific effect and the polygenic control of humoral and cellular responses also to R. equi.
Mutation in IRO1 Tightly Linked with URA3 Gene Reduces Virulence of Candida albicans
Authors: Chibana Hiroji , Uno Jun , Cho Tamaki , Mikami Yuzuru 
Published: November 2013

AbstractGene deletion in the pathogenic fungus Candida albicans has relied heavily on a URA3 cassette and a recipient Aura3 strain CAI4. The IRO1 gene adjacent to URA3 was inadvertently deleted during construction of CAI4. We report here that a mutation in IRO1 reduces virulence of C. albicans.
stx2vha Is the Dominant Genotype of Shiga Toxin‐Producing Escherichia coli O157:H7 Isolated from Patients and Domestic Animals in Three Regions of China
Authors: Zheng Han , Jing Huaiqi , Wang Hua , Xia Shengli , Hu Wangfu , Cui Shuyu , Bi Zhenqiang , Yang Jinchuan , Pang Bo , Zhao Guangfa , Zhang Jin , Li Hongwei , Xu Jianguo 
Published: November 2013

AbstractShiga toxin‐producing Escherichia coli (STEC) O157:H7 strains were isolated from domestic animals and patients from Xuzhou City, Jiangsu Province, China and the bordering Anhui and Henan Provinces and were examined for the stx genotype. Of 390 strains, 277 were identified as genotype stx2vha; 41, stx2; 51, stx2‐stx1; 1, stx2‐stx2vha‐stx1; 5, stx2‐stx2vha; and 15 were un‐typeable. Of the 277 stx2vha‐bearing isolates, 116 were isolated from goats; 42, cattle; 38, hens, and 35 from pigs. The study shows stx2vha is the dominant genotype and goats are an important reservoir.
Evaluation of the Biofilm‐Forming Ability and Genetic Typing for Clinical Isolates of Pseudomonas aeruginosa by Enterobacterial Repetitive Intergenic Consensus‐Based PCR
Authors: Yang Weiqing , Shi Lei , Jia Wen‐xiang , Yin Xiaolin , Su Jian‐yu , Kou Yali , Yi Xie , Shinoda Sumio , Miyoshi Shin‐ichi 
Published: November 2013

AbstractBiofilm formation is an important phenotype associated with chronic Pseudomonas aeruginosa infections. In the present study, a total of 48 P. aeruginosa strains isolated from clinical specimens were examined for their biofilm‐forming ability using a microtiter plate method. The different biofilm‐forming abilities were demonstrated among the strains; however, most strains formed a larger biofilm than strain PAO1, a reference strain. The genetic typing was also carried out by enterobacterial repetitive intergenic consensus‐based polymerase chain reaction. Although they were divided into five groups (A to E), most of the strains showing the higher biofilm‐forming ability were found to be in groups D and E, suggesting a significant relationship between the biofilm‐forming ability and the genetic group.
Protective Efficacy of Nonpathogenic Nef‐Deleted SHIV Vaccination Combined with Recombinant IFN‐γ Administration against a Pathogenic SHIV Challenge in Rhesus Monkeys
Authors: Kaneyasu Kentaro , Kita Masakazu , Ohkura Sadayuki , Yamamoto Toshiro , Ibuki Kentaro , Enose Yoshimi , Sato Akihiko , Kodama Makoto , Miura Tomoyuki , Hayami Masanori 
Published: November 2013

AbstractWe previously reported that a nef‐deleted SHIV (SHIV‐NI) is nonpathogenic and gave macaques protection from challenge infection with pathogenic SHIV‐C2/1. To investigate whether IFN‐γ augments the immune response induced by this vaccination, we examined the antiviral and adjuvant effect of recombinant human IFN‐γ (rIFN‐γ) in vaccinated and unvaccinated monkeys. Nine monkeys were vaccinated with nef‐deleted nonpathogenic SHIV‐NI. Four of them were administered with rIFN‐γ and the other five monkeys were administered with placebo. After the challenge with pathogenic SHIV‐C2/1, CD4+ T‐cell counts were maintained similarly in monkeys of both groups, while those of the unvaccinated monkeys decreased dramatically at 2 weeks after challenge. However, the peaks of plasma viral load were reduced to 100‐fold in SHIV‐NI vaccinated monkeys combined with rIFN‐γ compared with those in SHIV‐NI vaccinated monkeys without rIFN‐γ. The peaks of plasma viral load were inversely correlated with the number of SIV Gag‐specific IFN‐γ‐producing cells. In SHIV‐NI‐vaccinated monkeys with rIFN‐γ, the number of SIV Gag‐specific IFN‐γ‐producing cells of PBMCs increased 2‐fold compared with those in SHIV‐NI‐vaccinated monkeys without rIFN‐γ, and the NK activity and MIP‐1α production of PBMCs were also enhanced. Thus, vaccination of SHIV‐NI in combination with rIFN‐γ was more effective in modulating the antiviral immune system into a Th1 type response than SHIV‐NI vaccination alone. These results suggest that IFN‐γ augmented the anti‐viral effect by enhancing innate immunity and shifting the immune response to Th1.
Multicenter Evaluation of Commercial Frozen Plates for Microdilution Broth Antifungal Susceptibility Testing of Yeasts and Comparison of MIC Limits Recommended in NCCLS M27–A2
Authors: Makimura Koichi , Oguri Toyoko , Mikami Yuzuru , Kume Hikaru , Hanazawa Ryo , Abe Michiko , Ikeda Reiko , Shinoda Takako 
Published: November 2013

AbstractA commercial kit, Frozen Plate for Antifungal Susceptibility Testing of Yeasts, Eiken (Eiken Chemical Co., Ltd., Tokyo), was tested in a multi‐institute study to evaluate the agreement between inter‐institute MICs and National Committee for Clinical Laboratory Standards (NCCLS) M27–A2 recommendation limits of MIC value. The kit was reported as a method equivalent to the standardized guidelines for antifungal susceptibility testing by the Committee for Clinical Laboratory Standards—1994, the Japanese Society for Medical Mycology, and which is widely used in Japan for amphotericin B, flucytosine, fluconazole, miconazole, and itraconazole. The degrees of inter‐institute and NCCLS agreements were good to excellent especially with 48‐hr incubation for all antifungal agents. However, the percent agreements to NCCLS recommendations against itraconazole were poor. Overall, MIC values obtained using the frozen plate antifungal susceptibility testing kit, with 48‐hr incubation, were thought to be reliable and convenient alternatives to the data obtained by the NCCLS M27–A2 reference macrodilution and microdilution method. This kit will allow matching of results between international laboratories. However, the MIC value for itraconazole requires careful interpretation.
Nasal Peptide Vaccination Elicits CD8 Responses and Reduces Viral Burden after Challenge with Virulent Murine Cytomegalovirus
Authors: Gopal Indulekha N. , Quinn Anita , Henry Stanley C. , Hamilton John D. , Staats Herman F. , Frothingham Richard 
Published: November 2013

AbstractInfection of BALB/c mice with murine cytomegalovirus (MCMV) leads to CD8 cell responses to an immunodominant epitope YPHFMPTNL. We presented this epitope as a nasal peptide vaccine in combination with cholera toxin adjuvant, and evaluated immune responses and protection from MCMV challenge. Vaccination of naïve mice generated elevated numbers of peptide‐specific interferon‐7‐secreting splenocytes (median 80/million, range 60 to 490), compared to control mice (median 2/million, range —4.5 to 8; P=0.008, Mann‐Whitney test). Twelve days after challenge with virulent MCMV, vaccinated mice had a 1.1 log10 reduction in salivary gland viral titer compared to unvaccinated controls (5.36±0.24 vs. 6.42±0.12, mean±SD log10 plaque‐forming‐units; P<0.001, t‐test). Mice with chronic MCMV infection had consistent responses to the peptide (183±24/million interferon‐γ‐secreting splenocytes). Nasal peptide vaccination during chronic infection boosted peptide‐specific responses in two of four mice to >900/million interferon‐γ‐secreting splenocytes. Nasal peptide vaccination was immunogenic in naïve and MCMV‐infected mice, and reduced viral burden in naïve mice after virulent MCMV challenge. The nasal route may be useful for peptide presentation by novel human vaccines.
Interleukin‐4 Up‐Regulates T‐Tropic Human Immunodeficiency Virus Type 1 Transcription in Primary CD4+ CD38+ T‐Lymphocyte Subset
Authors: Li Yong‐Gang , Iwabu Yukie , Warachit Jiranan , Kinomoto Masanobu , Ibrahim Madiha S. , Tsuji Shoutaro , Mukai Tetsu , Kameoka Masanori , Tokunaga Kenzo , Sata Tetsutaro , Ikuta Kazuyoshi 
Published: November 2013

AbstractThe capacity of human immunodeficiency virus type 1 (HIV‐1) to infect resting cells and to produce progeny particles may contribute significantly to its pathogenicity in vivo. We previously reported that primary culture of resting CD4+ CD38+ T‐lymphocyte subset had higher production rate of CXCR4‐using (X4) HIV‐1 than CD4+ CD38− subset. Interleukin (IL)‐4 highly contributed to the up‐regulation of the X4 virus production in the CD38+ subset. Here, we show evidences that IL‐4 treatment of both resting CD38+ and CD38− subsets allowed the adsorption, entry, and integration of X4 virus at similar rates, while the following viral transcription rate was significantly lower in the CD38− than CD38+ subset. Treatment of the CD38 subsets with IL‐4 or phytohemagglutinin revealed no association of X4 virus replication ability in the subsets with classic T‐cell activation or proliferation. Interestingly, the activator protein (AP)‐1 was significantly activated in the CD38+ subset after IL‐4 treatment, while both nuclear factor (NF)‐κB and signal transducers and activator of transcription (STAT)‐6 were activated in the IL‐4‐treated CD38+ and CD38− subsets at similar levels. Thus, IL‐4‐dependent X4 HIV‐1 transcription occurs efficiently in the CD38+ but not CD38− subset of CD4+ population and AP‐1 could play a significant role on viral transcription, leading to the up‐regulated X4 virus production in the CD38+ subset.
Experimental Candidosis and Recovery of Candida albicans from the Oral Cavity of Ovariectomized Rats
Authors: Junqueira Juliana Campos , Colombo Carlos Eduardo Dias , Martins Joyce da Silva , Ito Cristiane Yumi Koga , Carvalho Yasmin Rodarte , Jorge Antonio Olavo Cardoso 
Published: November 2013

AbstractThe aim of this study was to analyze the development of candidosis and the recovery of C. albicans from the oral cavity of ovariectomized and sham‐ovariectomized rats. One hundred and twenty‐four rats originally negative for Candida spp. in the oral cavity were divided into two groups: ovariectomized and sham‐ovariectomized. Fifty‐eight ovariectomized and the same quantity of sham‐ovariectomized rats were inoculated with C. albicans for the study of candidosis development and recovery of yeast. Four animals from each group were not inoculated with yeast suspension and were submitted to tongue dorsum morphologic analysis by optical and scanning electron microscopy. The development of candidosis in the tongue dorsum was observed by optical and scanning electron microscopy in the periods of 6 hr, 24 hr, 7 days and 15 days after the last inoculation. Recovery of C. albicans was performed by oral samples plating on Sabouraud agar after 1, 2, 5 and 7 days and progressively at each 15‐day interval until negative cultures for yeasts were obtained. The results were analyzed by Mann‐Whitney and Student's t tests. The tongue dorsum of sham‐ovariectomized and ovariectomized rats, not infected by Candida, presented normal aspect. Among the infected rats, the ovariectomized group showed less occurrence of candidosis lesions and lower recovery of C. albicans from the oral cavity in relation to the sham‐ovariectomized group. It could be concluded that candidosis was less frequent from the oral cavities of ovariectomized rats in relation to sham‐ovariectomized.
Remarkable Synergies between Baicalein and Tetracycline, and Baicalein and β‐Lactams against Methicillin‐Resistant Staphylococcus aureus
Authors: Fujita Mai , Shiota Sumiko , Kuroda Teruo , Hatano Tsutomu , Yoshida Takashi , Mizushima Tohru , Tsuchiya Tomofusa 
Published: November 2013

AbstractDuring the screening of compounds that potentiate the effect of antimicrobial agents against methicillin‐resistant Staphylococcus aureus (MRSA), we found that an extract of thyme (Thymus vulgaris L) leaves greatly reduced the minimum inhibitory concentration (MIC) of tetracycline against MRSA. We isolated the effective compound and identified it as baicalein (5, 6, 7‐trihydroxyflavone). One of the clinically isolated MRSA strains possessed tetK, a gene encoding active efflux pump for tetracycline. We examined the effect of baicalein on the efflux of tetracycline, using Escherichia coli KAM32/pTZ1252 carrying the tetK. The E. coli KAM32/pTZ1252 showed 8 to 16 times higher MIC than E. coli KAM32. We observed strong inhibition of transport of tetracycline by baicalein with membrane vesicles prepared from E. coli KAM32/pTZ1252. Baicalein also showed synergy with tetracycline in a MRSA strain that doesn't possess tetK, or with β‐lactams. Thus, mechanisms of the synergies seem to be versatile.
Serratia marcescens Gene Required for Surfactant Serrawettin W1 Production Encodes Putative Aminolipid Synthetase Belonging to Nonribosomal Peptide Synthetase Family
Authors: Li Hong , Tanikawa Taichiro , Sato Yohei , Nakagawa Yoji , Matsuyama Tohey 
Published: November 2013

AbstractSerrawettin W1 produced by Serratia marcescens is a surface active exolipid having various functions supporting behaviors of bacteria on surface environments. Through the genetic analyses of serrawettin‐less mutants of S. marcescens 274, the swrW gene encoding putative serrawettin W1 synthetase was identified. Homology analysis of the putative SwrW demonstrated the presence of condensation, adenylation, thiolation, and thioesterase domains which are characteristic for nonribosomal peptide synthetase (NRPS). NRPSs have been known as multi‐modular enzymes. Linear alignment of these modules specifying respective amino acids will enable peptide bond formation resulting in a specific amino acid sequence. Putative SwrW was uni‐modular NRPS specifying only L‐serine. Possible steps in this simple uni‐modular NRPS for biosynthesis of serrawettin W1 [cyclo‐(D‐3‐hydroxydecanoyl‐L‐seryl)2] were predicted by referring to the ingenious enzymatic activity of gramicidin S synthetase (multi‐modular NRPS) of Brevibacillus brevis.
Construction of Human Fab Library and Isolation of Monoclonal Fabs with Rabies Virus‐Neutralizing Ability
Authors: Ando Tadasuke , Yamashiro Tetsu , Takita‐Sonoda Yoshiko , Mannen Kazuaki , Nishizono Akira 
Published: November 2013

AbstractA combinatorial human Fab library was constructed using RNAs from peripheral blood lymphocytes of 6 rabies vaccine‐boosted volunteers using pComb3X phagemid vector. The size of the constructed library was approximately 7.0×107 Escherichia coli transformants. The library was selected against purified rabies virus (RV) virion or purified RV glycoprotein for isolation of phages displaying RV‐neutralizing human Fab antibody. Among 132 selected clones, two Fab preparations revealed neutralizing activities against RV strain CVS when assayed in the rapid fluorescent focus inhibition test (RFFIT). The Fab preparation EP5G3 exhibited neutralizing activity with an infected cell count reduction of 76% at a dilution of 1:2, and of 20% at a dilution of 1:4. The Fab preparation GD2D12 also exhibited neutralizing activity with a 57% reduction at 1:2 and 41% reduction at 1:4. In the co‐immunoprecipitation using strain CVS, the RV glycoprotein was precipitated in reactions with both Fab preparations. The RV neutralizing ability of the Fab preparations described in the study were not directly correlated with their binding specificity for RV antigens detected by ELISA.
Experimental Abscess Formation Caused by Human Dental Plaque
Authors: Okayama Hidehito , Nagata Emi , Ito Hiro‐O , Oho Takahiko , Inoue Masakazu 
Published: November 2013

AbstractHuman dental plaque consists of a wide variety of microorganisms, some of which are believed to cause systemic infections, including abscesses, at various sites in the body. To confirm this hypothesis experimentally, we examined the abscess‐forming ability of native dental plaque in mice, the microbial features of the infectious locus produced by the plaque, and the anti‐phagocytic property of microbial isolates. Aliquots of a suspension of supragingival dental plaque containing 6 × 106 colony‐forming unit of bacteria were injected subcutaneously into the dorsa of mice. Abscess formation was induced in 76 of 85 mice using ten different plaque samples. Thirteen microorganisms were isolated from pus samples aspirated from abscess lesions. The microbial composition of pus, examined in 17 of 76 abscesses, was very simple compared to that of the plaque sample that had induced the abscess. The majority of the isolates belonged to the Streptococcus anginosus group, normally a minor component of plaque samples. S. anginosus was the most frequently detected organism and the most prevalent in seven abscesses, and Streptococcus intermedius and Streptococcus constellatus were predominant in one and three abscess samples, respectively. Each isolate of S. anginosus group produced abscesses in mice, and heat‐treated supragingival dental plaque influenced the abscess‐forming ability of S. anginosus isolate. These isolates possessed a high anti‐phagocytic capacity against human polymorphonuclear leukocytes. Our results suggest that human supragingival dental plaque itself is a source of the infectious pathogens that cause abscess formation.
Development of a Monoclonal Antibody against Epstein‐Barr Virus Nuclear Antigen Leader Protein (EBNA‐LP) That Can Detect EBNA‐LP Expressed in P3HR1 Cells
Authors: Shaku Fumio , Matsuda Go , Furuya Rie , Kamagata Chinatsu , Igarashi Mie , Tanaka Michiko , Kanamori Mikiko , Nishiyama Yukihiro , Yamamoto Naoki , Kawaguchi Yasushi 
Published: November 2013

AbstractA mouse monoclonal antibody, LP4D3, was raised against purified Epstein‐Barr virus nuclear antigen leader protein (EBNA‐LP) fused to glutathione‐S‐transferase. The antibody detected endogenous and exogenous EBNA‐LP in immunoblotting, immunofluorescence and immunoprecipitation assays, and the epitope of the antibody was mapped in the W2 domain of EBNA‐LP. While another monoclonal antibody to EBNA‐LP, JF186, which is widely used for analyses of the viral protein, did not react with truncated forms of EBNA‐LP expressed in P3HR1 cells, as reported earlier, the LP4D3 antibody did. The LP4D3 antibody will be a useful tool for further studies of EBNA‐LP, especially investigations into the phenotypes of mutant EBNA‐LP expressed in P3HR1 cells.