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Sample records for cell strain m5

  1. Complete genome sequences of Brucella melitensis strains M28 and M5-90, with different virulence backgrounds.

    Science.gov (United States)

    Wang, Fangkun; Hu, Sen; Gao, Yuzhe; Qiao, Zujian; Liu, Wenxing; Bu, Zhigao

    2011-06-01

    Brucella melitensis is a Gram-negative coccobacillus bacteria belonging to the Alphaproteobacteria subclass. It is an important zoonotic pathogen that causes brucellosis, a disease affecting sheep, cattle, and sometimes humans. The B. melitensis strain M5-90, a live attenuated vaccine cultured from the B. melitensis virulent strain M28, has been an effective tool to control brucellosis in goats and sheep in China. Here we report the complete genome sequences of B. melitensis M28 and M5-90, strains with different virulence backgrounds, which will serve as a valuable reference for future studies.

  2. Long-term dissemination of CTX-M-5-producing hypermutable Salmonella enterica serovar typhimurium sequence type 328 strains in Russia, Belarus, and Kazakhstan.

    Science.gov (United States)

    Kozyreva, Varvara K; Ilina, Elena N; Malakhova, Maja V; Carattoli, Alessandra; Azizov, Ilya S; Tapalski, Dmitry V; Kozlov, Roman S; Edelstein, Mikhail V

    2014-09-01

    In this paper, we present evidence of long-term circulation of cefotaxime-resistant clonally related Salmonella enterica serovar Typhimurium strains over a broad geographic area. The genetic relatedness of 88 isolates collected from multiple outbreaks and sporadic cases of nosocomial salmonellosis in various parts of Russia, Belarus, and Kazakhstan from 1996 to 2009 was established by multilocus tandem-repeat analysis (MLVA) and multilocus sequence typing (MLST). The isolates belong to sequence type 328 (ST328) and produce CTX-M-5 β-lactamase, whose gene is carried by highly related non-self-conjugative but mobilizable plasmids. Resistance to nalidixic acid and low-level resistance to ciprofloxacin is present in 37 (42%) of the isolates and in all cases is determined by various single point mutations in the gyrA gene quinolone resistance-determining region (QRDR). Isolates of the described clonal group exhibit a hypermutable phenotype that probably facilitates independent acquisition of quinolone resistance mutations.

  3. Studies of the mechanism of the cyclisation reaction catalysed by the wildtype and a truncated alpha-cyclodextrin glycosyltransferase from Klebsiella pneumoniae strain M 5 al, and the beta-cyclodextrin glycosyltransferase from Bacillus circulans strain 8.

    Science.gov (United States)

    Bender, H

    1990-10-10

    The actions of the wildtype and a truncated alpha-cyclodextrin glycosyltransferase from Klebsiella pneumoniae strain M 5 al on malto-oligosaccharides showed no significant differences, and there was marked dependence of the kinetic parameters on the chain lengths of the substrate. The action of the beta-cyclodextrin glycosyltransferase from Bacillus circulans was less dependent on the chain length of the substrate, but Vmax of the initial cyclisation with the longer malto-oligosaccharides was only 28% of that determined for the enzyme of K. pneumoniae. The rate parameters suggested that the active site of each enzyme spans nine glucosyl residues, and that the catalytic sites are situated between subsites three and four for the K. pneumoniae enzymes and between subsites two and three for the B. circulans enzyme. The molecular binding affinities and the affinities of the 9th subsite were calculated from the rate parameters. The primary and tertiary structures of alpha-amylases and cyclodextrin glycosyltransferases are compared in the context of the reaction mechanism of the latter enzymes.

  4. Short communication: Antiproliferative effect of 8 different Lactobacillus strains on K562 cells.

    Science.gov (United States)

    Tuo, Yanfeng; Jiang, Shujuan; Qian, Fang; Mu, Guangqing; Liu, Peng; Guo, Yuanji; Ma, Changlu

    2015-01-01

    Some strains of Lactobacillus genus have antiproliferative activities against cancer cells. However, until now, the exact effector molecules of Lactobacillus strains with anticancer activity have not been identified. The aim of the present study was to explore which fraction of the Lactobacillus cells exerts the highest antiproliferative effect. For this purpose, the heat-killed bacterial cells, bacterial cell wall extract, and genomic DNA of 8 Lactobacillus strains were prepared to assess their antiproliferative activities against human myeloid leukemia cell lines K562. The heat-killed bacterial cells of the 8 lactobacilli strains exerted antiproliferative effect on K562 cells, and the inhibition rates exerted by the heat-killed bacterial cells of the strains G15AL, M5AL, SB31AL, SB5AL, and T3AL were significantly higher than those exerted by the cell walls and genomic DNA of the strains. The bacterial DNA of G15AL exerted higher antiproliferative effect on K562 cells. The exact effector molecules and the effect mechanism of the strains should be further explored for the application of these strains as probiotic strains or bioactive probiotic molecules.

  5. Effect of oxalioplatin on human colorectal cancer cell line LoVo and cell subline SW480/M5%奥沙利铂对人结肠癌细胞LoVo和SW480/M5作用的实验研究

    Institute of Scientific and Technical Information of China (English)

    吕会增; 魏波; 陈图峰; 苏雁甜; 叶小勇; 陈新岐; 卫洪波

    2012-01-01

    目的 探讨体外实验奥沙利铂对人结肠癌LoVo和SW480/M5细胞的抑制作用及其可能机制.方法 MTT法检测不同剂量奥沙利铂对LoVo和SW480/M5细胞增殖的抑制作用.流式细胞仪检测1/2 GI50和GI50浓度奥沙利铂对LoVo和SW480/M5细胞周期和早期凋亡的影响.原子光谱吸收仪检测GI50浓度奥沙利铂作用4、8、24 h后LoVo和SW480/M5细胞DNA含铂量.结果 奥沙利铂对LoVo和SW480/M5细胞增殖的抑制作用呈量效依赖;其GI50浓度:LoVo细胞为6.5 mg/L,SW480/M5细胞为58.0 mg/L.自然对数增殖周期,LoVo细胞中G1期细胞比例高于、G2期细胞比例低于SW480/M5细胞(P<0.05).奥沙利铂浓度GI50时,降低肿瘤细胞G1期比例,升高LoVo细胞S期比例较SW480/M5细胞明显,升高SW480/M5细胞而降低LoVo细胞G2/M期比例.1/2 GI50、GI50奥沙利铂均可诱导两种肿瘤细胞发生早期凋亡,但1/2 GI50 L-OHP促凋亡作用两种肿瘤细胞间无统计学差异,GI50 L-OHP对LoVo细胞的促凋亡作用高于SW480/M5细胞.GI50 L-OHP奥沙利铂使两种肿瘤细胞DNA含铂量显著升高,并呈时效依赖.LoVo细胞DNA交联铂原子能力高于SW480/M5细胞.结论 奥沙利铂主要通过阻滞细胞于S期或(和)G2/M期并诱导细胞凋亡而抑制两种肿瘤细胞增殖.LoVo细胞对奥沙利铂敏感性明显高于SW480/M5细胞.

  6. Cells as strain-cued automata

    Science.gov (United States)

    Cox, Brian N.; Snead, Malcolm L.

    2016-02-01

    We argue in favor of representing living cells as automata and review demonstrations that autonomous cells can form patterns by responding to local variations in the strain fields that arise from their individual or collective motions. An autonomous cell's response to strain stimuli is assumed to be effected by internally-generated, internally-powered forces, which generally move the cell in directions other than those implied by external energy gradients. Evidence of cells acting as strain-cued automata have been inferred from patterns observed in nature and from experiments conducted in vitro. Simulations that mimic particular cases of pattern forming share the idealization that cells are assumed to pass information among themselves solely via mechanical boundary conditions, i.e., the tractions and displacements present at their membranes. This assumption opens three mechanisms for pattern formation in large cell populations: wavelike behavior, kinematic feedback in cell motility that can lead to sliding and rotational patterns, and directed migration during invasions. Wavelike behavior among ameloblast cells during amelogenesis (the formation of dental enamel) has been inferred from enamel microstructure, while strain waves in populations of epithelial cells have been observed in vitro. One hypothesized kinematic feedback mechanism, "enhanced shear motility", accounts successfully for the spontaneous formation of layered patterns during amelogenesis in the mouse incisor. Directed migration is exemplified by a theory of invader cells that sense and respond to the strains they themselves create in the host population as they invade it: analysis shows that the strain fields contain positional information that could aid the formation of cell network structures, stabilizing the slender geometry of branches and helping govern the frequency of branch bifurcation and branch coalescence (the formation of closed networks). In simulations of pattern formation in

  7. On strain and stress in living cells

    Science.gov (United States)

    Cox, Brian N.; Smith, David W.

    2014-11-01

    Recent theoretical simulations of amelogenesis and network formation and new, simple analyses of the basic multicellular unit (BMU) allow estimation of the order of magnitude of the strain energy density in populations of living cells in their natural environment. A similar simple calculation translates recent measurements of the force-displacement relation for contacting cells (cell-cell adhesion energy) into equivalent volume energy densities, which are formed by averaging the changes in contact energy caused by a cell's migration over the cell's volume. The rates of change of these mechanical energy densities (energy density rates) are then compared to the order of magnitude of the metabolic activity of a cell, expressed as a rate of production of metabolic energy per unit volume. The mechanical energy density rates are 4-5 orders of magnitude smaller than the metabolic energy density rate in amelogenesis or bone remodeling in the BMU, which involve modest cell migration velocities, and 2-3 orders of magnitude smaller for innervation of the gut or angiogenesis, where migration rates are among the highest for all cell types. For representative cell-cell adhesion gradients, the mechanical energy density rate is 6 orders of magnitude smaller than the metabolic energy density rate. The results call into question the validity of using simple constitutive laws to represent living cells. They also imply that cells need not migrate as inanimate objects of gradients in an energy field, but are better regarded as self-powered automata that may elect to be guided by such gradients or move otherwise. Thus Ġel=d/dt 1/2 >[(C11+C12)ɛ02+2μγ02]=(C11+C12)ɛ0ɛ˙0+2μγ0γ˙0 or Ġel=ηEɛ0ɛ˙0+η‧Eγ0γ˙0 with 1.4≤η≤3.4 and 0.7≤η‧≤0.8 for Poisson's ratio in the range 0.2≤ν≤0.4 and η=1.95 and η‧=0.75 for ν=0.3. The spatial distribution of shear strains arising within an individual cell as cells slide past one another during amelogenesis is not known

  8. Adhesions of extracellular surface-layer associated proteins in Lactobacillus M5-L and Q8-L.

    Science.gov (United States)

    Zhang, Yingchun; Xiang, Xinling; Lu, Qianhui; Zhang, Lanwei; Ma, Fang; Wang, Linlin

    2016-02-01

    Surface-layer associated proteins (SLAP) that envelop Lactobacillus paracasei ssp. paracasei M5-L and Lactobacillus casei Q8-L cell surfaces are involved in the adherence of these strain to the human intestinal cell line HT-29. To further elucidate some of the properties of these proteins, we assessed the yields and expressions of SLAP under different incubation conditions. An efficient and selective extraction of SLAP was obtained when cells of Lactobacillus were treated with 5 M LiCl at 37°C in aerobic conditions. The SLAP of Lactobacillus M5-L and Q8-L in cell extracts were visualized by SDS-PAGE and identified by Western blotting with sulfo-N-hydroxysuccinimide-biotin-labeled HT-29 cells as adhesion proteins. Atomic force microscopy contact imaging revealed that Lactobacillus strains M5-L and Q8-L normally display a smooth, homogeneous surface, whereas the surfaces of M5-L and Q8-L treated with 5 M LiCl were rough and more heterogeneous. Analysis of adhesion forces revealed that the initial adhesion forces of 1.41 and 1.28 nN obtained for normal Lactobacillus M5-L and Q8-L strains, respectively, decreased to 0.70 and 0.48 nN, respectively, following 5 M LiCl treatment. Finally, the dominant 45-kDa protein bands of Lactobacillus Q8-L and Lactobacillus M5-L were identified as elongation factor Tu and surface antigen, respectively, by liquid chromatography-tandem mass spectrometry.

  9. Supersymmetric Perturbations of the M5 brane

    CERN Document Server

    Niarchos, Vasilis

    2014-01-01

    We study long-wavelength supersymmetric deformations of brane solutions in supergravity using an extension of previous ideas within the general scheme of the blackfold approach. As a concrete example, we consider long-wavelength perturbations of the planar M2-M5 bound state solution in eleven-dimensional supergravity. We propose a specific ansatz for the first order deformation of the supergravity fields and explore how this deformation perturbs the Killing spinor equations. We find that a special part of these equations gives a projection equation on the Killing spinors that has the same structure as the $\\kappa$-symmetry condition of the abelian M5 brane theory. Requiring a match between supergravity and gauge theory implies a specific non-linear gauge-gravity map between the bosonic fields of the abelian M5 brane theory and the gravity-induced fluid-like degrees of freedom of the blackfold equations that control the perturbative gravity solution. This observation sheds new light on the SUGRA/DBI correspond...

  10. Mycobacterium tuberculosis strains exhibit differential and strain-specific molecular signatures in pulmonary epithelial cells.

    Science.gov (United States)

    Mvubu, Nontobeko Eunice; Pillay, Balakrishna; Gamieldien, Junaid; Bishai, William; Pillay, Manormoney

    2016-12-01

    Although pulmonary epithelial cells are integral to innate and adaptive immune responses during Mycobacterium tuberculosis infection, global transcriptomic changes in these cells remain largely unknown. Changes in gene expression induced in pulmonary epithelial cells infected with M. tuberculosis F15/LAM4/KZN, F11, F28, Beijing and Unique genotypes were investigated by RNA sequencing (RNA-Seq). The Illumina HiSeq 2000 platform generated 50 bp reads that were mapped to the human genome (Hg19) using Tophat (2.0.10). Differential gene expression induced by the different strains in infected relative to the uninfected cells was quantified and compared using Cufflinks (2.1.0) and MeV (4.0.9), respectively. Gene expression varied among the strains with the total number of genes as follows: F15/LAM4/KZN (1187), Beijing (1252), F11 (1639), F28 (870), Unique (886) and H37Rv (1179). A subset of 292 genes was commonly induced by all strains, where 52 genes were down-regulated while 240 genes were up-regulated. Differentially expressed genes were compared among the strains and the number of induced strain-specific gene signatures were as follows: F15/LAM4/KZN (138), Beijing (52), F11 (255), F28 (55), Unique (186) and H37Rv (125). Strain-specific molecular gene signatures associated with functional pathways were observed only for the Unique and H37Rv strains while certain biological functions may be associated with other strain signatures. This study demonstrated that strains of M. tuberculosis induce differential gene expression and strain-specific molecular signatures in pulmonary epithelial cells. Specific signatures induced by clinical strains of M. tuberculosis can be further explored for novel host-associated biomarkers and adjunctive immunotherapies.

  11. Cytopathic effect of PPR vaccine virus strains in Vero cells

    Directory of Open Access Journals (Sweden)

    Raveendra Hegde

    2009-06-01

    Full Text Available The present study describes the cytopathic effect of two different Peste des petits ruminants (PPR vaccine virus strains presently being used in the country, in vero cells. The cytopathic effect (CPE was visible from 4th day post infection in Sungri vaccine virus strain where as Arasur vaccine virus strain showed CPE, 36-48 hr post infection. With both vaccine virus strains the CPE in vero cells showed initial cell rounding, aggregation and syncytial development. The generalized CPE was noticed by 6th day in Sungri and by 96 hrs post infection in Arasur strain. However complete detachment of the cell monolayer was observed in Arasur strain by 120 hr, post infection. Infected coverslip cultures stained with H & E and May & Grunwald’s Giemsa showed cell vaculation, cytoplasmic extension and syncytia comprising of five to six nuclei. Acidophilic intracytoplasmic and intranuclear inclusion bodies were also observed. Titers, HA activity and detection by s-ELISA of both the vaccine virus strains are also compared. [Vet. World 2009; 2(3.000: 93-94

  12. Effect of Magnetic Field on L-Strain Cells

    CERN Document Server

    Ulakoglu, G; Atak, C; Rzakoulieva, A; Danilov, V I; Alikamanoglu, S

    2000-01-01

    The effects of electromagnetic and magnetic fields are currently being made useful in many fields, especially in medicine. In this research work, L-Strain cells which are a type of fibrosarcoma cells were exposed to a magnetic flow of 2-26 mT in periods of 1, 2, 3 and 4 minutes. The L-Strain cells, which were exposed to the magnetic field for these periods, were counted after 24 and 48 hours, when compared with the controls, it was observed that in groups of 1 and 4 minutes exposure a significant decrease (P < 0.05) in the number of cells occurred. The per cent of labelling index of L-Strain cells exposed to the magnetic field for 1 and 4 minutes decreased significantly also in comparison to the controls.

  13. Survey of radiosensitivity in a variety of human cell strains

    Energy Technology Data Exchange (ETDEWEB)

    Arlett, C.F.; Harcourt, S.A.

    1980-03-01

    Gamma-ray sensitivity for cell killing was assayed in 54 human cell strains, including some derived from individuals suffering from certain hereditary diseases. The overall range of Do values in this study was 38 to 180 rads, indicating a considerable range of variability in humans. The normal sensitivity was described by a range of Do values of 97 to 180 rads. All ten ataxia telangiectasia cell strains tested proved radiosensitive and gave a mean Do value of 57 +- 15 (S.E.) rads, and these represent the most radiosensitive human skin fibroblasts currently available. Representative cell strains from familial retinoblastoma, Fanconi's anemia, and Hutchinson-Gilford progeria occupied positions of intermediate sensitivity, as did one of two ataxia telangiectasia heterozygotes. Six xeroderma pigmentosum cell strains together with two Cockayne's syndrome cell strains (all known to be sensitive to ultraviolet light) fell into the normal range, indicating an absence of cross-sensitivity between ultraviolet light and gamma-irradiation.

  14. Stretching Behavior of Red Blood Cells at High Strain Rates

    Science.gov (United States)

    Mancuso, Jordan; Ristenpart, William

    2016-11-01

    Most work on the mechanical behavior of red blood cells (RBCs) has focused on simple shear flows. Relatively little work has examined RBC deformations in the physiologically important extensional flow that occurs at the entrance to a constriction. In particular, previous work suggests that RBCs rapidly stretch out and then retract upon entering the constriction, but to date no model predicts this behavior for the extremely high strain rates typically experienced there. In this work, we use high speed video to perform systematic measurements of the dynamic stretching behavior of RBCs as they enter a microfluidic constriction. We demonstrate that a simple viscoelastic model captures the observed stretching dynamics, up to strain rates as high as 1000 s-1. The results indicate that the effective elastic modulus of the RBC membrane at these strain rates is an order of magnitude larger than moduli measured by micropipette aspiration or other low strain rate techniques.

  15. Cell Factory Stability and Genetic Circuits for Improved Strain Development

    DEFF Research Database (Denmark)

    Rugbjerg, Peter

    Development of new chemical-­‐producing microbial cell factories is an iterative trial-­and-­error process, and to screen candidate cells at high throughput, genetic biosensor systems are appealing. Each biosensor has distinct biological parameters, making modular tuning networks attractive....... However, all synthetic gene systems -­ including the target metabolic pathways themselves -­ represent a possible fitness burden to the cell and thus constitute a threat to strain stability. In this thesis, several studies served to develop genetic systems for optimizing cell factory development...... factories in future....

  16. Invasion of Porphyromonas gingivalis strains into vascular cells and tissue

    Directory of Open Access Journals (Sweden)

    Ingar Olsen

    2015-08-01

    Full Text Available Porphyromonas gingivalis is considered a major pathogen in adult periodontitis and is also associated with multiple systemic diseases, for example, cardiovascular diseases. One of its most important virulence factors is invasion of host cells. The invasion process includes attachment, entry/internalization, trafficking, persistence, and exit. The present review discusses these processes related to P. gingivalis in cardiovascular cells and tissue. Although most P. gingivalis strains invade, the invasion capacity of strains and the mechanisms of invasion including intracellular trafficking among them differ. This is consistent with the fact that there are significant differences in the pathogenicity of P. gingivalis strains. P. gingivalis invasion mechanisms are also dependent on types of host cells. Although much is known about the invasion process of P. gingivalis, we still have little knowledge of its exit mechanisms. Nevertheless, it is intriguing that P. gingivalis can remain viable in human cardiovascular cells and atherosclerotic plaque and later exit and re-enter previously uninfected host cells.

  17. Orthopoxvirus species and strain differences in cell entry

    Energy Technology Data Exchange (ETDEWEB)

    Bengali, Zain; Satheshkumar, P.S. [Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-3210 (United States); Moss, Bernard, E-mail: bmoss@nih.gov [Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-3210 (United States)

    2012-11-25

    Vaccinia virus (VACV) enters cells by a low pH endosomal route or by direct fusion with the plasma membrane. We previously found differences in entry properties of several VACV strains: entry of WR was enhanced by low pH, reduced by bafilomycin A1 and relatively unaffected by heparin, whereas entry of IHD-J, Copenhagen and Elstree were oppositely affected. Since binding and entry modes may have been selected by specific conditions of in vitro propagation, we now examined the properties of three distinct, recently isolated cowpox viruses and a monkeypox virus as well as additional VACV and cowpox virus strains. The recent isolates were more similar to WR than to other VACV strains, underscoring the biological importance of endosomal entry by orthopoxviruses. Sequence comparisons, gene deletions and gene swapping experiments indicated that viral determinants, other than or in addition to the A26 and A25 'fusion-suppressor' proteins, impact entry properties.

  18. A PURE STRAIN OF CARTILAGE CELLS IN VITRO.

    Science.gov (United States)

    Fischer, A

    1922-09-30

    1. A strain of cartilage cells, obtained from the pars cartilago sclerae of the eye of chick embryos, has been cultivated for more than 3 months in vitro. 2. The initial growth of the cartilage was possible only on the free surface of the coagulum. 3. The hyaline substance disappeared during cultivation in vitro. The succeeding stages of a transformation from small, lymphocyte-like cells into large, spindle-shaped cells were observed. The cartilage cells were spindle-shaped and grew in close contact, forming thin membranes. In surface-grown cartilage cells, the nucleus, usually containing one large nucleolus, stained less deeply than the cytoplasm. 4. The rate of growth of cartilage was slower than that of fibroblasts and epithelium. After cultivation on the surface of the coagulum, the cartilage cells could multiply even when embedded in the coagulum. But their growth was less extensive and uniform.

  19. Emergency therapeutic leukapheresis in a case of acute myeloid leukemia M5

    Science.gov (United States)

    Ranganathan, Sudha; Sesikeran, Shyamala; Gupta, Vineet; Vanajakshi

    2008-01-01

    Cell separators in India are routinely used for plateletpheresis, peripheral blood stem cell collections and therapeutic plasma exchange. Therapeutic leukapheresis, particularly as an emergency procedure, has been uncommonly performed and reported. Here, a case of a 53-year-old male, diagnosed with acute myeloid leukemia subtype M5 (AML M5) with hyperleukocytosis, who underwent emergency leukaphereis, is reported. After two procedures, there was a decrease of WBC count by 85%, which enabled cytotoxic therapy to be initiated. PMID:20041073

  20. Role for M5 muscarinic acetylcholine receptors in cocaine addiction.

    Science.gov (United States)

    Fink-Jensen, Anders; Fedorova, Irina; Wörtwein, Gitta; Woldbye, David P D; Rasmussen, Thøger; Thomsen, Morgane; Bolwig, Tom G; Knitowski, Karen M; McKinzie, David L; Yamada, Masahisa; Wess, Jürgen; Basile, Anthony

    2003-10-01

    Muscarinic cholinergic receptors of the M5 subtype are expressed by dopamine-containing neurons of the ventral tegmentum. These M5 receptors modulate the activity of midbrain dopaminergic neurons, which play an important role in mediating reinforcing properties of abused psychostimulants like cocaine. The potential role of M5 receptors in the reinforcing effects of cocaine was investigated using M5 receptor-deficient mice in a model of acute cocaine self-administration. The M5-deficient mice self-administered cocaine at a significantly lower rate than wild-type controls. In the conditioned place preference procedure, a classic test for evaluating the rewarding properties of drugs, M5-deficient mice spent significantly less time in the cocaine-paired compartment than control mice. Moreover, the severity of the cocaine withdrawal syndrome (withdrawal-associated anxiety measured in the elevated plus-maze) was significantly attenuated in mice lacking the M5 receptor. These results demonstrate that M5 receptors play an important role in mediating both cocaine-associated reinforcement and withdrawal.

  1. 布鲁氏菌M5-90疫苗株磷酸葡萄糖变位酶基因(pgm)突变株的构建及免疫评价%Construction and Immune Evaluation on Mutation of phosphoglucomutase gene (pgm) Attenuates of Brucella Melitensis Vaccine M5-90

    Institute of Scientific and Technical Information of China (English)

    李天森; 张辉; 张艳; 孟茹; 张豫; 孙志华; 蒋攀文; 王震; 陈瑞花

    2013-01-01

    In order to obtain a Brucella vaccine candidate which can distinguish between natural infection and attenuated vaccine, this paper studied the phosphoglucomutase gene (pgm) influence on Brucella M5-90 vaccine strain virulence and immune evaluation. The recombinant plasmid pGEM-7zf-Δpgm was constructed, and which was electroporated into Brucella melitensis M5-90 competent cells, screening for Brucella vaccine strain M5-90 pgm gene deletion mutant strains (Δpgm) and to detect the M5-90 Δpgm strain genetic stability, and immunogenicity. The test results showed that the reverse mutation did not occur within 15 passages; the Δpgm antibody content and the parent strain M5-90 group had no significant difference; Δpgm had the ability to produce the IL-2 was stronger than that of M5-90, the ability to produce INF-γ was lower than the M5-90; the rose bengal plate agglutination test and tube agglutination test of this gene deletion strains was not agglutinated; the results of Western blot confirmed that the Δpgm could not induce the mice to produce anti-pgm protein antibody. The Δpgm constructed in this study, had good genetic stability and immunogenicity, provided basic data for the next step to more in-depth develop Brucella gene-deleted vaccine.%为获得可以区分自然感染和疫苗免疫且毒力较弱的布鲁氏菌候选疫苗株,本实验研究了磷酸葡萄糖变位酶基因(pgm)对布鲁氏菌(Brucella melitensis)M5-90疫苗株毒力的影响,并对其进行了免疫评价.本实验构建了重组质粒pGEM-7zf-△pgm,电转化布鲁氏菌M5-90感受态细胞,筛选获得布鲁氏菌疫苗株M5-90的pgm基因缺失株(△pgm),并对获得的M5-90 △pgm株进行遗传稳定性、免疫原性检测.结果显示,△pgm能稳定传15代;△pgm组的抗体含量较亲本株M5-90组差异不显著;△pgm在诱导机体产生IL-2的能力要强于M5-90,产生INF-γ的能力低于M5-90;该基因缺失株采用虎红平板凝集试验和试管凝集

  2. Increased amphetamine-induced locomotor activity, sensitization, and accumbal dopamine release in M5 muscarinic receptor knockout mice

    DEFF Research Database (Denmark)

    Schmidt, Lene S; Miller, Anthony D; Lester, Deranda B

    2010-01-01

    showed that M(5) receptor knockout (M (5) (-/-) ) mice are less sensitive to the reinforcing properties of addictive drugs. MATERIALS AND METHODS: Here, we investigate the role of M(5) receptors in the effects of amphetamine and cocaine on locomotor activity, locomotor sensitization, and dopamine release...... using M (5) (-/-) mice backcrossed to the C57BL/6NTac strain. STATISTICAL ANALYSES: Sensitization of the locomotor response is considered a model for chronic adaptations to repeated substance exposure, which might be related to drug craving and relapse. The effects of amphetamine on locomotor activity......-induced hyperactivity and dopamine release as well as amphetamine sensitization are enhanced in mice lacking the M(5) receptor. These results support the concept that the M(5) receptor modulates effects of addictive drugs....

  3. Elliptical posts allow for detailed control of non-equibiaxial straining of cell cultures

    DEFF Research Database (Denmark)

    Olesen, Christian Gammelgaard; Pennisi, Cristian Pablo; de Zee, Mark

    2013-01-01

    Background A modification of the Flexcell system that allows imposition of homogenous, controlled non-equibiaxial strains to cell cultures is developed and experimentally validated. The Flexcell system by default applies equibiaxial strain to cell cultures, meaning no shear strain, while soft tis...

  4. The sinusoidal periodicity nature for M>=5 global earthquakes

    CERN Document Server

    Zhang, Z X

    2016-01-01

    By using the M>=5 global earthquake data for Jan. 1950 to Dec. 2015, we performed statistical analyses for the parameters magnitude, time, and depth on a yearly scale. The magnitude spectrum, which is the earthquake number accumulated at different magnitudes, had an exponential distribution. For the first time, we report a very significant characteristic of the sinusoidal periodic variation in the spectral index. The cycle of the sine function fitting was 30.98 years. The concept of annual equivalent total magnitude (AETM) of total released energy for each year was introduced and the trend variation of AETM year by year was studied. Overall, the global AETM of earthquakes with M>=5 displayed a certain upward trend as the years elapsed. At the same time, the change of the average epicenter depth of the global earthquakes (M>=5) in each year was analyzed.

  5. Cytopathogenesis of Naegleria fowleri Thai strains for cultured human neuroblastoma cells.

    Science.gov (United States)

    Tiewcharoen, Supathra; Malainual, Nat; Junnu, Virach; Chetanachan, Pruksawan; Rabablert, Jundee

    2008-04-01

    The aim of this study is to evaluate cellular interaction between free-living amoebae Naegleria fowleri strains and mammalian target cells in vitro. Two Thai strains of N. fowleri; Khon Kaen strain from the environment and Siriraj strain from the patient's cerebrospinal fluid and the Center of Disease Control VO 3081 strain from Atlanta (US) were studied. Human neuroblastoma (SK-N-MC) and African Green monkey Kidney (Vero) cells were used as target cells. Each cell line was inoculated with each strain of N. fowleri at a ratio of 1:1 and observed for 7 days. The uninoculated target cells and each strain of N. fowleri were used as control. The numbers of the challenged and unchallenged cells as well as the free-living amoebae were counted three times by trypan blue exclusion method. The inoculation began when the amoebae attached to the cell membrane and ingested the target cells. In this study, extensive cytopathogenesis with many floating inoculated cells and abundant number of amoebae were observed. The destruction pattern of both inoculated SK-N-MC and Vero target cells were similar. Interestingly, SK-N-MC was more susceptible to N. fowleri strains than the Vero cell. In addition, N. fowleri Siriraj strain showed the highest destruction pattern for each target cell. Our findings suggest that the SK-N-MC should be used as a base model for studying the neuropathogenesis in primary amoebic meningoencephalitis patients.

  6. The Blue Straggler Population of the Globular Cluster M5

    CERN Document Server

    Lanzoni, B; Ferraro, F R; Mancini, C; Beccari, G; Rood, R T; Mapelli, M; Sigurdsson, S

    2007-01-01

    By combining high-resolution HST and wide-field ground based observations, in ultraviolet and optical bands, we study the Blue Stragglers Star (BSS) population of the galactic globular cluster M5 (NGC 5904) from its very central regions up to its periphery. The BSS distribution is highly peaked in the cluster center, decreases at intermediate radii and rises again outward. Such a bimodal distribution is similar to those previously observed in other globular clusters (M3, 47Tucanae, NGC6752). As for these clusters, dynamical simulations suggest that, while the majority of BSS in M5 could be originated by stellar collisions, a significant fraction (20-40%) of BSS generated by mass transfer processes in primordial binaries is required to reproduce the observed radial distribution. A candidate BSS has been detected beyond the cluster tidal radius. If confirmed, this could represent an interesting case of an "evaporating" BSS.

  7. Wilson Loops for M2- and M5-brane spaces

    CERN Document Server

    Quijada, Edward

    2015-01-01

    We calculate the quark and anti-quark interaction energy in different positions in spaces generated by $N$ coincident $M2$- and $M5$-branes. We use the Maldacena-Rey-Yee method for calculating this energy as a function of quark-antiquark separation. We obtain the solution for these problems as integrals of the metric elements. For limiting regimes we find simpler solutions for which some potentials exhibit a confinement behavior.

  8. Supersymmetric attractors, topological strings, and the M5-brane CFT

    Science.gov (United States)

    Guica, Monica M.

    One of the purposes of this thesis is to present the consistent and unifying picture that emerges in string and M-theory with eight supercharges. On one hand, this involves classifying and relating supersymmetric objects that occur in N = 2 compactifications of string and M-theory on a Calabi-Yau manifold. These come in a surprisingly wide variety of four and five-dimensional black holes, black rings and their sometimes very complicated bound states. On the other hand, the topological string also makes its appearance in theories with eight supercharges, and turns out to compute certain black hole degeneracies. We dedicate the introduction and the first chapter to summarizing and reviewing the beautiful relationships between black holes, black rings, their dual conformal field theory and the topological string, and we also outline the remaining puzzles and issues. Some of the black holes in question can be obtained by multiply-wrapping an M-theory M5-brane on a self-intersecting four-cycle in the Calabi-Yau manifold. Their dual microscopic description is known, and consists of a two-dimensional conformal field theory (CFT) which is the low-energy limit of the gauge theory that resides on the worldvolume of the M5 brane. We show that in a certain limit the M5-brane CFT is - perhaps surprisingly - able to reproduce the entropy of a completely different type of black holes, those obtained from wrapped M2-branes, whose microscopic description has not yet been understood. We also argue that certain black hole bound states should also be described by the same CFT, which suggests a unifying description of the various black objects in eight-supercharge supergravity theories. Finally, we describe and present a proof of the so-called OSV conjecture, which states that the mixed partition function of N = 2 four-dimensional BPS black holes equals the modulus square of the type A topological string partition function. We also attempt to use this relationship to better understand

  9. The geometry of the M5-branes and TQFTs

    CERN Document Server

    Bonelli, G

    2001-01-01

    The calculation of the partition function for N M5-branes is addressed for the case in which the worldvolume wraps a manifold $T^2\\times M_4$, where $M_4$ is simply connected and Kaehler. This is done in a compactification of M-theory which induces the Vafa-Witten theory on $M_4$ in the limit of vanishing torus volume. The results follow from the equivalence of the BPS spectrum counting in the complementary limit of vanishing $M_4$ volumes and from a classification of the the moduli space of quantum vacua of the supersymmetric twisted theory in terms of associated spectral covers. This reduces the problem of the moduli counting to algebraic equations.

  10. 3D supergravity from wrapped M5-branes

    CERN Document Server

    Karndumri, Parinya

    2015-01-01

    Through consistent Kaluza-Klein reduction, we construct 3D N=2 gauged supergravities corresponding to twisted compactifications of M5-branes on a product of Riemann surfaces, including Kahler-Einstein four-manifolds. We extend the reduction to fermionic supersymmetry variations in order to determine the 3D Killing spinor equations and classify all (timelike) supersymmetric solutions. We show that the superpotential T dictates all supersymmetric solutions, not just AdS3 vacua. As a by-product, we identify an infinite class of new supersymmetric warped AdS3 (Godel) and warped dS3 critical points. Moreover, we show that T encodes the central charge and R symmetry of the dual N = (0,2) SCFTs in the large N limit. Upon uplift to 11D, we use this result to write the higher-dimensional geometries in canonical form and discuss the relation to existing classifications of supersymmetric AdS3 geometries.

  11. M3与M5的MICM鉴别

    Institute of Scientific and Technical Information of China (English)

    索翠平; 李寅; 赵霞

    2003-01-01

    @@ 急性早幼粒细胞自血病(M3型,acute promyelocytic leukemia,APL)和急性单核细胞白血病(M5型,acute monocytic leukemia,AMOL)同属于急性非淋巴细胞白血病.二者在临床与形态上均有相似之处,特别是细颗粒型急性早幼粒细胞白血病(M3b)在形态上更易被误诊为急性单核细胞白血病.但二者的治疗与预后却有很大的区别.在此,结合MICM分型原则对二者加以鉴别,以免造成误诊,导致不应有的后果.

  12. Groundwater Level Prediction using M5 Model Trees

    Science.gov (United States)

    Nalarajan, Nitha Ayinippully; Mohandas, C.

    2015-01-01

    Groundwater is an important resource, readily available and having high economic value and social benefit. Recently, it had been considered a dependable source of uncontaminated water. During the past two decades, increased rate of extraction and other greedy human actions have resulted in the groundwater crisis, both qualitatively and quantitatively. Under prevailing circumstances, the availability of predicted groundwater levels increase the importance of this valuable resource, as an aid in the planning of groundwater resources. For this purpose, data-driven prediction models are widely used in the present day world. M5 model tree (MT) is a popular soft computing method emerging as a promising method for numeric prediction, producing understandable models. The present study discusses the groundwater level predictions using MT employing only the historical groundwater levels from a groundwater monitoring well. The results showed that MT can be successively used for forecasting groundwater levels.

  13. 3D supergravity from wrapped M5-branes

    Science.gov (United States)

    Karndumri, Parinya; Ó Colgáin, Eoin

    2016-03-01

    Through consistent Kaluza-Klein reduction, we construct 3D N=2 gauged supergravities corresponding to twisted compactifications of M5-branes on a product of constant curvature Riemann surfaces, including Kähler-Einstein four-manifolds. We extend the reduction to fermionic supersymmetry variations in order to determine the 3D Killing spinor equations and classify all timelike supersymmetric solutions. As a by-product, we identify an infinite class of new supersymmetric warped AdS 3 (Gödel) and warped dS 3 solutions. Moreover, we show that the superpotential T encodes the central charge and R symmetry of the dual N=(0,2) SCFTs in the large N limit. We demonstrate that the R symmetry matches the canonical U(1) isometry from existing classifications of supersymmetric AdS 3 solutions to 11D supergravity with N=(0,2) supersymmetry.

  14. Sensitivity to fuel diesel oil and cell wall structure of some Scenedesmus (Chlorococcales strains

    Directory of Open Access Journals (Sweden)

    Zbigniew Tukaj

    2014-01-01

    Full Text Available Sensitivity of three Scenedesmus strains exposed to aqueous fuel-oil extract (AFOE is strongly strain-dependent S. quadricauda is the most resistant, S. armatus moderately tolerant whereas the most sensitive appears to be S. microspina. The sensitivity of tested species increases parallel with decreasing of cell size and cell number in coenobium. The values of the cell surface/cell volumes ratios only partly explain the above relationships. Electron microscope investigations reveal that the sensitivity may depend on cell wall structure of the strains. Cell wall of all here investigated strains is built of two layers: the inner so-called cellulosic layer and the outer one showing a three-laminar structure (TLS. The latter contains an acetolysis-resistant biopolymer (ARB. These two layers are similar in thickness in the three strains tested, but the surface of Scenedesmus is covered with various epistructures that are characteristic of strains. Some of them as the tightly fitting warty layer of S. armatus and especially the loosely fitting reticulate layer of S. quadricauda may contribute to lower permeability of cell wall. The structure of the rosettes also appears to be correlated with the sensitivity of strains. Presence of invaginations of plasmalemma in areas under rosettes indicates their role in transport processes inside/outside the cells.

  15. Effects of equiaxial strain on the differentiation of dental pulp stem cells without using biochemical reagents.

    Science.gov (United States)

    Tabatabaei, F S; Jazayeri, M; Ghahari, P; Haghighipour, N

    2014-09-01

    During orthodontic treatments, applied mechanical forces create strain and result in tooth movement through the alveolar bone. This response to mechanical strain is a fundamental biological reaction. The present study evaluated the effect of equiaxial strain within the range of orthodontic forces on the osteogenic differentiation of human dental pulp stem cells (hDPSCs). Following isolation and culture of hDPSCs, 3rd passage cells were transferred on a silicone membrane covered with collagen. Cell adhesion to the membrane was evaluated under scanning electron microscope (SEM). Cells were divided into three groups: the first group was placed in a conventional culture medium, transferred to an equiaxial stretching device (3% strain for 2 weeks). The positive control was placed in an osteogenic medium with no mechanical strain. The negative control group was placed in the conventional culture medium with no mechanical strain either. Study groups were evaluated for expression ofosteogenic markers (Alkaline phosphatase and Osteopontin) with immunofluorescence and real time PCR. SEM images revealed optimal adhesion of cells to the silicone membrane. Immunofluorescence study demonstrated that osteocalcin expression occurred after 2 weeks in the two groups under mechanical and chemical signals. After application of equiaxial strain, level of expression of osteogenic markers was significantly higher than in the negative and positive control groups. Based on the study results, static equiaxial strain which mimics the types of orthodontic forces can result in differentiation of hDPSCs to osteoblasts. The results obtained may be used in cell therapy and tissue engineering.

  16. Interaction of Aeromonas strains with lactic acid bacteria via Caco-2 cells.

    Science.gov (United States)

    Hatje, E; Neuman, C; Katouli, M

    2014-01-01

    The genus Aeromonas includes some species that have now been identified as human pathogens of significant medical importance. We investigated the ability of 13 selected Aeromonas strains belonging to nine species isolated from clinical cases (n = 5), environmental waters (n = 5), and fish (n = 3) to adhere to and translocate Caco-2 cells in the absence and presence of two lactic acid bacteria (LAB), i.e., Lactobacillus acidophilus and Bifidobacterium breve. Aeromonas isolates were also assessed for their cytotoxicity, the presence of virulence genes, and hemolysin production. Among the clinical isolates, one strain of Aeromonas veronii biovar veronii and two strains of Aeromonas hydrophila carried cytotoxin (act), heat-labile toxin (alt), hemolysin (hlyA), and aerolysin (aerA) genes, were cytotoxic to Vero cells, produced hemolysin, and showed higher adherence to Caco-2 cells. In contrast, this was seen in only one environmental strain, a strain of A. veronii biovar sobria. When Aeromonas strains were coinoculated with LAB onto Caco-2 cells, their level of adhesion was reduced. However, their rate of translocation in the presence of LAB increased and was significantly (P Aeromonas and LAB strains could have a detrimental effect on the Caco-2 cells, allowing the Aeromonas to translocate more readily, or the presence of the LAB stimulated the Aeromonas strains to produce more toxins and/or increase their translocation rate.

  17. A Massive Neutron Star in the Globular Cluster M5

    CERN Document Server

    Freire, Paulo C C; Berg, Maureen van den; Hessels, Jason W T

    2007-01-01

    We report the results of 18 years of Arecibo timing of two pulsars in the globular cluster NGC 5904 (M5), PSR B1516+02A and PSR B1516+02B. This has allowed the measurement of the proper motions of these pulsars and of the cluster. PSR B1516+02B is a 7.95-ms pulsar in a binary system with a ~0.2 solar-mass companion and an orbital period of 6.86 days. In deep HST images, no optical counterpart is detected at the position of the pulsar, implying the companion is either a white dwarf or a low-mass MS star. The eccentricity of the orbit (e = 0.14) has allowed a measurement of the rate of advance of periastron: 0.0136 +/ 0.0007 degrees per year. It is very likely that the periastron advance is due to the effects of general relativity; the total mass of the binary system is then 2.14 +/- 0.16 solar masses. The small measured mass function implies, in a statistical sense, that a very large fraction of this total mass is contained in the pulsar: 1.94 +0.17/-0.19 solar masses (1 sigma$); there is a 5 % probability tha...

  18. Precision radial velocities of 15 M5 - M9 dwarfs

    CERN Document Server

    Barnes, J R; Jones, H R A; Jeffers, S V; Rojo, P; Arriagada, P; Jordan, A; Minniti, D; Tuomi, M; Pinfield, D; Anglada-Escude, G

    2014-01-01

    We present radial velocity measurements of a sample of M5V-M9V stars from our Red-Optical Planet Survey, ROPS, operating at 0.65-1.025 micron. Radial velocities for 15 stars, with r.m.s. precision down to 2.5 m/s over a week long time scale are achieved using Thorium-Argon reference spectra. We are sensitive to planets with m_psin(i) >= 1.5 MEarth (3 MEarth at 2-sigma) in the classical habitable zone and our observations currently rule out planets with m_psin(i) > 0.5 MJup at 0.03 AU for all our targets. A total of 9 of the 15 targets exhibit r.m.s. 10 MEarth in 0.03 AU orbits. Since the mean rotation velocity is of order 8 km/s for an M6V star and 15 km/s by M9V, we avoid observing only slow rotators that would introduce a bias towards low axial inclination i << 90 deg systems, which are unfavourable for planet detection. Our targets with the highest vsini values exhibit radial velocities significantly above the photon-noise limited precision, even after accounting for vsini. We therefore monitored st...

  19. Supersymmetric M5 brane theories on R × CP2

    Science.gov (United States)

    Kim, Hee-Cheol; Lee, Kimyeong

    2013-07-01

    We propose 4 and 12 supersymmetric conformal Yang-Mills-Chern-Simons theories on R × CP2 as multiple representations of the theory on M5 branes. These theories are obtained by twisted Zk modding and dimensional reduction of the 6d (2,0) superconformal field theory on R × S5 and have a discrete coupling constant 1/{g_{{YM}^2}}=k/{4{π^2}} with natural number k. Instantons in these theories are expected to represent the Kaluza-Klein modes. For the k = 1 , 2 cases, we argue that the number of supersymmetries in our theories should be enhanced to 32 and 16, respectively. For the k = 3 case, only the 4 supersymmetric theory gets the supersymmetric enhancement to 8. For the 4 supersymmetric case, the vacuum structure becomes more complicated as there are degenerate supersymmetric vacua characterized by fuzzy spheres. We calculate the perturbative part of the SU( N ) gauge group Euclidean path integral for the index function at the symmetric phase of the 4 supersymmetric case and confirm it with the known half-BPS index. From the similar twisted Z k modding of the AdS7 × S4 geometry, we speculate that the M region is for k ≲ N 1/3 and the type IIA region is N 1/3 ≲ k ≲ N. When nonperturbative corrections are included, our theories are expected to produce the full index of the 6d (2,0) theory.

  20. In vitro permissivity of bovine cells for wild-type and vaccinal myxoma virus strains.

    Science.gov (United States)

    Pignolet, Béatrice; Duteyrat, Jean-Luc; Allemandou, Aude; Gelfi, Jacqueline; Foucras, Gilles; Bertagnoli, Stéphane

    2007-09-27

    Myxoma virus (MYXV), a leporide-specific poxvirus, represents an attractive candidate for the generation of safe, non-replicative vaccine vector for non-host species. However, there is very little information concerning infection of non-laboratory animals species cells with MYXV. In this study, we investigated interactions between bovine cells and respectively a wild type strain (T1) and a vaccinal strain (SG33) of MYXV. We showed that bovine KOP-R, BT and MDBK cell lines do not support MYXV production. Electron microscopy observations of BT-infected cells revealed the low efficiency of viral entry and the production of defective virions. In addition, infection of bovine peripheral blood mononuclear cells (PBMC) occurred at a very low level, even following non-specific activation, and was always abortive. We did not observe significant differences between the wild type strain and the vaccinal strain of MYXV, indicating that SG33 could be used for new bovine vaccination strategies.

  1. Biocatalytic Resolution of para-Nitrostyrene Oxide by Resting Cells of Different Aspergillus niger Strains

    Institute of Scientific and Technical Information of China (English)

    金浩; 李祖义; 王清

    2001-01-01

    Biocatalytic resolution of racemic para-nitrostyrene oxide was accomplished by employing the epoxide hydrolases from the whole cells of several Aspergillus niger (A. niger) strains. In the cases investigated, excellent selectivity was achieved with such strains as A, niger 5450, A. niger 5320.

  2. Cloning and expression of a toxin gene from Pseudomonas fluorescens GcM5-1A.

    Science.gov (United States)

    Kong, Lingying; Guo, Daosen; Zhou, Shiyi; Yu, Xinlei; Hou, Guixue; Li, Ronggui; Zhao, Boguang

    2010-07-01

    Pseudomonas fluorescens GcM5-1A was isolated from the pine wood nematode (PWN), Bursaphelenchus xylophilus, obtained from wilted Japanese black pine, Pinus thumbergii, in China. In this paper, a genomic library of the GcM5-1A strain was constructed and a toxin-producing clone was isolated by bioassay. Nucleotide sequence analysis revealed an open reading frame of 1,290 bp encoding a protein of 429 amino acids with N-terminal putative signal peptide of 36 amino acids, which shared a similarity of 83, 82 and 80% identity with hypothetical protein PFLU2919 from P. fluorescens SBW25, Dyp-type peroxidase family protein from P. fluorescens Pf-5 and Tat-translocated enzyme from P. fluorescens Pf0-1, respectively. The gene encoding a full-length protein or without the putative signal peptide was cloned and expressed as a soluble protein in E. coli. The recombinant protein was purified to electrophoretic homogeneity by affinity chromatography using a Ni2+ matrix column. Its relative molecular weight was estimated to be 48.5 kDa by SDS-PAGE for full-length protein, and 45.0 kDa for the recombinant protein without putative signal peptide. Bioassay results showed that the recombinant protein with or without the putative signal peptide was toxic to both suspension cells and P. thunbergii seedlings. HPLC analysis demonstrated that components in branch extracts of P. thunbergii were significantly changed after addition of the recombinant full-length protein and hydrogen peroxide, which indicated that it is probably a peroxidase. This study offers information that can be used to determine the mechanism of pine wilt disease caused by the PWN.

  3. BCG strain S4-Jena: An early BCG strain is capable to reduce the proliferation of bladder cancer cells by induction of apoptosis

    Directory of Open Access Journals (Sweden)

    Hermann Inge-Marie

    2010-06-01

    Full Text Available Abstract Background Intravesical immunotherapy with Mycobacterium bovis bacillus Calmette-Guérin has been established as the most effective adjuvant treatment for high risk non-muscle-invasive bladder cancer (NMIBC. We investigated the differences between the S4-Jena BCG strain and commercially available BCG strains. We tested the genotypic varieties between S4-Jena and other BCG strains and analysed the effect of the BCG strains TICE and S4-Jena on two bladder cancer cell lines. Results In contrast to commercially available BCG strains the S4-Jena strain shows genotypic differences. Spoligotyping verifies the S4-Jena strain as a BCG strain. Infection with viable S4-Jena or TICE decreased proliferation in the T24 cell line. Additionally, hallmarks of apoptosis were detectable. In contrast, Cal29 cells showed only a slightly decreased proliferation with TICE. Cal29 cells infected with S4-Jena, though, showed a significantly decreased proliferation in contrast to TICE. Concordantly with these results, infection with TICE had no effect on the morphology and hallmarks of apoptosis of Cal29 cells. However, S4-Jena strain led to clearly visible morphological changes and caspases 3/7 activation and PS flip. Conclusions S4-Jena strain has a direct influence on bladder cancer cell lines as shown by inhibition of cell proliferation and induction of apoptosis. The data implicate that the T24 cells are responder for S4-Jena and TICE BCG. However, the Cal29 cells are only responder for S4-Jena and they are non-responder for TICE BCG. S4-Jena strain may represent an effective therapeutic agent for NMIBC.

  4. Strain-specific differences in pili formation and the interaction of Corynebacterium diphtheriae with host cells

    Directory of Open Access Journals (Sweden)

    Hensel Michael

    2010-10-01

    Full Text Available Abstract Background Corynebacterium diphtheriae, the causative agent of diphtheria, is well-investigated in respect to toxin production, while little is known about C. diphtheriae factors crucial for colonization of the host. In this study, we investigated strain-specific differences in adhesion, invasion and intracellular survival and analyzed formation of pili in different isolates. Results Adhesion of different C. diphtheriae strains to epithelial cells and invasion of these cells are not strictly coupled processes. Using ultrastructure analyses by atomic force microscopy, significant differences in macromolecular surface structures were found between the investigated C. diphtheriae strains in respect to number and length of pili. Interestingly, adhesion and pili formation are not coupled processes and also no correlation between invasion and pili formation was found. Using RNA hybridization and Western blotting experiments, strain-specific pili expression patterns were observed. None of the studied C. diphtheriae strains had a dramatic detrimental effect on host cell viability as indicated by measurements of transepithelial resistance of Detroit 562 cell monolayers and fluorescence microscopy, leading to the assumption that C. diphtheriae strains might use epithelial cells as an environmental niche supplying protection against antibodies and macrophages. Conclusions The results obtained suggest that it is necessary to investigate various isolates on a molecular level to understand and to predict the colonization process of different C. diphtheriae strains.

  5. Holographic cosmology from a system of M2–M5 branes

    Energy Technology Data Exchange (ETDEWEB)

    Sepehri, Alireza, E-mail: alireza.sepehri@uk.ac.ir [Faculty of Physics, Shahid Bahonar University, P.O. Box 76175, Kerman (Iran, Islamic Republic of); Research Institute for Astronomy and Astrophysics of Maragha (RIAAM), Maragha (Iran, Islamic Republic of); Faizal, Mir, E-mail: f2mir@uwaterloo.ca [Department of Physics and Astronomy, University of Waterloo, Waterloo, ON, N2L 3G1 (Canada); Setare, Mohammad Reza, E-mail: rezakord@ipm.ir [Department of Science, Campus of Bijar, University of Kurdistan, Bijar (Iran, Islamic Republic of); Ali, Ahmed Farag, E-mail: afali@fsu.edu [Department of Physics, Florida State University, Tallahassee, FL 32306 (United States); Department of Physics, Faculty of Science, Benha University, Benha 13518 (Egypt)

    2016-05-15

    In this paper, we analyze the holographic cosmology using a M2–M5 brane configuration. In this configuration, a M2-brane will be placed in between a M5-brane and an anti-M5-brane. The M2-brane will act as a channel for energy to flow from an anti-M5-brane to a M5-brane, and this will increase the degrees of freedom on the M5-brane causing inflation. The inflation will end when the M5-brane and anti-M5-brane get separated. However, at a later stage the distance between the M5-brane and the anti-M5-bran can reduce and this will cause the formation of tachyonic states. These tachyonic states will again open a bridge between the M5-branes and the anti-M5-branes, which will cause further acceleration of the universe.

  6. Holographic cosmology from a system of M2-M5 branes

    Science.gov (United States)

    Sepehri, Alireza; Faizal, Mir; Setare, Mohammad Reza; Ali, Ahmed Farag

    2016-05-01

    In this paper, we analyze the holographic cosmology using a M2-M5 brane configuration. In this configuration, a M2-brane will be placed in between a M5-brane and an anti-M5-brane. The M2-brane will act as a channel for energy to flow from an anti-M5-brane to a M5-brane, and this will increase the degrees of freedom on the M5-brane causing inflation. The inflation will end when the M5-brane and anti-M5-brane get separated. However, at a later stage the distance between the M5-brane and the anti-M5-bran can reduce and this will cause the formation of tachyonic states. These tachyonic states will again open a bridge between the M5-branes and the anti-M5-branes, which will cause further acceleration of the universe.

  7. Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands.

    Directory of Open Access Journals (Sweden)

    Eri O Maruyama

    Full Text Available The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER was targeted to the prolactin-induced protein (Pip gene locus. Targeting of the Dcpp1 gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the Tcfcp2l1 gene locus. Using the R26Tomato Red reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using PipGCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing DcppGCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts.

  8. Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands.

    Science.gov (United States)

    Maruyama, Eri O; Aure, Marit H; Xie, Xiaoling; Myal, Yvonne; Gan, Lin; Ovitt, Catherine E

    2016-01-01

    The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER) was targeted to the prolactin-induced protein (Pip) gene locus. Targeting of the Dcpp1 gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the Tcfcp2l1 gene locus. Using the R26Tomato Red reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using PipGCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing DcppGCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts.

  9. Raspberry wine fermentation with suspended and immobilized yeast cells of two strains of Saccharomyces cerevisiae.

    Science.gov (United States)

    Djordjević, Radovan; Gibson, Brian; Sandell, Mari; de Billerbeck, Gustavo M; Bugarski, Branko; Leskošek-Čukalović, Ida; Vunduk, Jovana; Nikićević, Ninoslav; Nedović, Viktor

    2015-01-01

    The objectives of this study were to assess the differences in fermentative behaviour of two different strains of Saccharomyces cerevisiae (EC1118 and RC212) and to determine the differences in composition and sensory properties of raspberry wines fermented with immobilized and suspended yeast cells of both strains at 15 °C. Analyses of aroma compounds, glycerol, acetic acid and ethanol, as well as the kinetics of fermentation and a sensory evaluation of the wines, were performed. All fermentations with immobilized yeast cells had a shorter lag phase and faster utilization of sugars and ethanol production than those fermented with suspended cells. Slower fermentation kinetics were observed in all the samples that were fermented with strain RC212 (suspended and immobilized) than in samples fermented with strain EC1118. Significantly higher amounts of acetic acid were detected in all samples fermented with strain RC212 than in those fermented with strain EC1118 (0.282 and 0.602 g/l, respectively). Slightly higher amounts of glycerol were observed in samples fermented with strain EC1118 than in those fermented with strain RC212.

  10. Zika Virus Strains Potentially Display Different Infectious Profiles in Human Neural Cells

    Directory of Open Access Journals (Sweden)

    Yannick Simonin

    2016-10-01

    Full Text Available The recent Zika virus (ZIKV epidemic has highlighted the poor knowledge on its physiopathology. Recent studies showed that ZIKV of the Asian lineage, responsible for this international outbreak, causes neuropathology in vitro and in vivo. However, two African lineages exist and the virus is currently found circulating in Africa. The original African strain was also suggested to be neurovirulent but its laboratory usage has been criticized due to its multiple passages. In this study, we compared the French Polynesian (Asian ZIKV strain to an African strain isolated in Central African Republic and show a difference in infectivity and cellular response between both strains in human neural stem cells and astrocytes. Consistently, this African strain led to a higher infection rate and viral production, as well as stronger cell death and anti-viral response. Our results highlight the need to better characterize the physiopathology and predict neurological impairment associated with African ZIKV.

  11. [Composition of cell walls of 2 mutant strains of Streptomyces chrysomallus].

    Science.gov (United States)

    Zaretskaia, M Sh; Nefelova, M V; Baratova, L A; Polin, A N

    1984-12-01

    The cell walls and peptidoglycans of two mutant strains, Streptomyces chrysomallus var. carotenoides and Streptomyces chrysomallus var. macrotetrolidi, were studied. The strains are organisms producing carotenes and antibiotics of the macrotetrolide group. By the qualitative composition of the peptidoglycans the mutants belong to Streptomyces and are similar. Their glycan portion consists of equimolar quantities of N-acetyl glucosamine and muramic acid. The peptide subunit is presented by glutamic acid, L, L-diaminopimelic acid, glycine and alanine. The molar ratio of alanine is 1.2-1.3. The mutant strains differ in the content of carbohydrates, total phosphorus and phosphorus belonging to teichoic acids. Teichoic acids of the cell walls of the both strains are of the ribitolhosphate nature. The cell walls of the mutants contain polysaccharides differing from teichoic acids and consisting of glucose, galactose, arabinose and fucose. The influence of the cell wall composition of the mutant strains on their morphology and metabolism and comparison of the data relative to the mutant strains with those relative to the starting strain are discussed.

  12. 荧光假单胞菌GcM51 A重组鞭毛蛋白的表达及其对黑松细胞的毒性研究%Expression of Recombinant Flagellin from Pseudomonas Fluorescens GcM5 1 A and It’s Toxicity to Pinus Thunbergii Cells

    Institute of Scientific and Technical Information of China (English)

    肖莉; 张蕾; 牟卉; 李荣贵

    2013-01-01

    针对松萎蔫病的早期诊断及生物防治,本文利用PCR 技术从荧光假单胞菌 GcM51 A的基因组中克隆了鞭毛蛋白编码基因 fliC,再将该基因克隆到表达载体 pET 15 b的NcoI 和XhoI位点,构建重组质粒pET 15 b fliC,再将重组质粒导入大肠杆菌BL21(DE3),构建工程菌,IPTG诱导工程菌高效表达 C 末端具有多聚组氨酸标签的重组鞭毛蛋白,重组蛋白主要以包涵体的形式存在,包涵体经8 mol/L尿素溶解,复性并经 Ni2+螯合柱亲和层析得到了电泳纯的重组鞭毛蛋白。生测结果表明,重组鞭毛蛋白可引起黑松细胞的大量死亡,与天然鞭毛蛋白对黑松愈伤组织细胞有相似的毒性。该研究为松萎蔫病致病机理的研究奠定了基础。%In this paper,gene fliC encoding flagellin was amplified from genomic DNA of Pseudomonasflu-orescens GcM5 1A by polymerase chain reaction (PCR),and cloned into pET 15b to construct recombi-nant plasmid pET 15b fliC.This plasmid was introduced into Escherichiacoli BL21(DE3)to construct engineering bacteria,and overexpression of recombinant flagellin with a his-tag at its C-terminal was a-chieved when engineering bacteria were induced by isopropylβ D 1 thiogalactopyranoside (IPTG). The recombinant protein in engineering bacteria appeared mainly in the form of inclusion bodies.Recombi-nant flagellin in inclusion bodies,which was dissolved by 8 mol/L urea and refolded,was purified through affinity chromatography on a nickel column.Bioassay results indicated that the recombinant protein was toxic to callus cells of Pinus thunbergii,which had a similar result to native flagellin.This study laid a good foundation for further research on the mechanism of pine wilt disease.

  13. Planar cell polarity aligns osteoblast division in response to substrate strain.

    Science.gov (United States)

    Galea, Gabriel L; Meakin, Lee B; Savery, Dawn; Taipaleenmaki, Hanna; Delisser, Peter; Stein, Gary S; Copp, Andrew J; van Wijnen, Andre J; Lanyon, Lance E; Price, Joanna S

    2015-03-01

    Exposure of bone to dynamic strain increases the rate of division of osteoblasts and also influences the directional organization of the cellular and molecular structure of the bone tissue that they produce. Here, we report that brief exposure to dynamic substrate strain (sufficient to rapidly stimulate cell division) influences the orientation of osteoblastic cell division. The initial proliferative response to strain involves canonical Wnt signaling and can be blocked by sclerostin. However, the strain-related orientation of cell division is independently influenced through the noncanonical Wnt/planar cell polarity (PCP) pathway. Blockade of Rho-associated coiled kinase (ROCK), a component of the PCP pathway, prevents strain-related orientation of division in osteoblast-like Saos-2 cells. Heterozygous loop-tail mutation of the core PCP component van Gogh-like 2 (Vangl2) in mouse osteoblasts impairs the orientation of division in response to strain. Examination of bones from Vangl2 loop-tail heterozygous mice by µCT and scanning electron microscopy reveals altered bone architecture and disorganized bone-forming surfaces. Hence, in addition to the well-accepted role of PCP involvement in response to developmental cues during skeletal morphogenesis, our data reveal that this pathway also acts postnatally, in parallel with canonical Wnt signaling, to transduce biomechanical cues into skeletal adaptive responses. The simultaneous and independent actions of these two pathways appear to influence both the rate and orientation of osteoblast division, thus fine-tuning bone architecture to meet the structural demands of functional loading.

  14. Membrane rigidity of red blood cells parasitized by different strains of Plasmodium falciparum.

    Science.gov (United States)

    Paulitschke, M; Nash, G B

    1993-11-01

    Changes in the structure of parasitized red blood cells may influence their ability to circulate. We have used a micropipette technique to examine the effects of invasion and maturation of Plasmodium falciparum on the membrane rigidity of red blood cells. In the presence of immature, ring form parasites from different laboratory strains, membrane rigidity remained unchanged as compared with uninfected red cells. However, development of more mature pigmented trophozoites caused a marked increase in membrane rigidity. Parasites from knobless strains caused a less-pronounced increase than parasites from knob-positive strains. Using closely synchronized cultures, the dependence of membrane rigidity on parasite maturation was studied in more detail for selected knob-positive and knobless strains. Over a period of 12 hours, while trophozoites developed into schizonts, no further rigidification of the red cell membrane occurred. The increase in membrane rigidity, occurring with the initial development of pigmented trophozoites, may be related to insertion of neoantigens into the red cell surface or modification of native membrane proteins that also occur at this time. In contrast to others, we found no effect of parasite-culture supernatant, harvested at different stages, on the rigidity of uninfected cells exposed to it. Interstrain variation of membrane rigidity could influence pathophysiology in several ways: by promoting margination and cytoadherence of knob-positive strains in the microcirculation, by modulating clearance of parasitized cells by the reticuloendothelial system, and by influencing ischemic complications of severe falciparum malaria.

  15. Finite-Element Modeling of Viscoelastic Cells During High-Frequency Cyclic Strain

    Directory of Open Access Journals (Sweden)

    David W. Holdsworth

    2012-03-01

    Full Text Available Mechanotransduction refers to the mechanisms by which cells sense and respond to local loads and forces. The process of mechanotransduction plays an important role both in maintaining tissue viability and in remodeling to repair damage; moreover, it may be involved in the initiation and progression of diseases such as osteoarthritis and osteoporosis. An understanding of the mechanisms by which cells respond to surrounding tissue matrices or artificial biomaterials is crucial in regenerative medicine and in influencing cellular differentiation. Recent studies have shown that some cells may be most sensitive to low-amplitude, high-frequency (i.e., 1–100 Hz mechanical stimulation. Advances in finite-element modeling have made it possible to simulate high-frequency mechanical loading of cells. We have developed a viscoelastic finite-element model of an osteoblastic cell (including cytoskeletal actin stress fibers, attached to an elastomeric membrane undergoing cyclic isotropic radial strain with a peak value of 1,000 µstrain. The results indicate that cells experience significant stress and strain amplification when undergoing high-frequency strain, with peak values of cytoplasmic strain five times higher at 45 Hz than at 1 Hz, and peak Von Mises stress in the nucleus increased by a factor of two. Focal stress and strain amplification in cells undergoing high-frequency mechanical stimulation may play an important role in mechanotransduction.

  16. Finite-element modeling of viscoelastic cells during high-frequency cyclic strain.

    Science.gov (United States)

    Milner, Jaques S; Grol, Matthew W; Beaucage, Kim L; Dixon, S Jeffrey; Holdsworth, David W

    2012-03-22

    Mechanotransduction refers to the mechanisms by which cells sense and respond to local loads and forces. The process of mechanotransduction plays an important role both in maintaining tissue viability and in remodeling to repair damage; moreover, it may be involved in the initiation and progression of diseases such as osteoarthritis and osteoporosis. An understanding of the mechanisms by which cells respond to surrounding tissue matrices or artificial biomaterials is crucial in regenerative medicine and in influencing cellular differentiation. Recent studies have shown that some cells may be most sensitive to low-amplitude, high-frequency (i.e., 1-100 Hz) mechanical stimulation. Advances in finite-element modeling have made it possible to simulate high-frequency mechanical loading of cells. We have developed a viscoelastic finite-element model of an osteoblastic cell (including cytoskeletal actin stress fibers), attached to an elastomeric membrane undergoing cyclic isotropic radial strain with a peak value of 1,000 µstrain. The results indicate that cells experience significant stress and strain amplification when undergoing high-frequency strain, with peak values of cytoplasmic strain five times higher at 45 Hz than at 1 Hz, and peak Von Mises stress in the nucleus increased by a factor of two. Focal stress and strain amplification in cells undergoing high-frequency mechanical stimulation may play an important role in mechanotransduction.

  17. Adhesion of Two Lactobacillus gasseri Probiotic Strains on Caco-2 Cells

    Directory of Open Access Journals (Sweden)

    Mojca Narat

    2003-01-01

    Full Text Available Previous in vitro and in vivo studies showed that two human isolates of Lactobacillus gasseri, LF221 and K7 are able to survive the passage through the gastrointestinal tract and to colonise intestines of pigs at least temporarily. The aim of this study was to examine the adhesion ability of LF221 and K7 strains to Caco-2 cells. Adhesion of lactobacilli from early stationary growth phase was examined at two pH values of DMEM buffer (4.5 and 7. Lactobacillus rhamnosus GG, a widely used strain with clinical evidences of its efficiency, served as a positive control. The number of lactobacilli added to each well was found to be crucial in the adhesion assay. When added, lactobacilli were in range of 2.5 · 106 to 2.5 · 108 cfu/well, the linear correlation between the number of adhered cells (log cfu and the number of added cells (log cfu was found for all three strains (R2 > 0.99 at both pH values (4.5 and 7. At the highest concentration of added K7 and GG cells tested (app. 109 cfu/well, the efficiency of adhesion was reduced. pH value of the medium strongly affected the adhesion, which was promoted in acidic conditions (pH=4.5. The adhesion of K7 strain was slightly weaker compared to GG strain at both pH values, while at pH=4.5 the adhesion of LF221 strain was even better than GG adhesion, at least at lower concentration of lactobacilli. The direct comparison of these strains was possible by regression analysis. At lower concentration of lactobacilli (2.5 · 106, the best efficiency of adhesion (% of adhered bacteria was observed for the strain LF221, reaching the values of 7.8 and 1.9 % at pH=4.5 and 7, respectively, while at higher lactobacilli concentration the ration of adhesion was higher for GG strain (3.3 % at pH=4.5. In conclusion, strains LF221 and K7 were demonstrated to be adhesive, especially in acidic conditions. The level of adhesion of K7 and GG strains positively correlates with the number of added lactobacilli only up to the

  18. Inefficient complement system clearance of Trypanosoma cruzi metacyclic trypomastigotes enables resistant strains to invade eukaryotic cells.

    Directory of Open Access Journals (Sweden)

    Igor Cestari

    Full Text Available The complement system is the main arm of the vertebrate innate immune system against pathogen infection. For the protozoan Trypanosoma cruzi, the causative agent of Chagas disease, subverting the complement system and invading the host cells is crucial to succeed in infection. However, little attention has focused on whether the complement system can effectively control T. cruzi infection. To address this question, we decided to analyse: 1 which complement pathways are activated by T. cruzi using strains isolated from different hosts, 2 the capacity of these strains to resist the complement-mediated killing at nearly physiological conditions, and 3 whether the complement system could limit or control T. cruzi invasion of eukaryotic cells. The complement activating molecules C1q, C3, mannan-binding lectin and ficolins bound to all strains analysed; however, C3b and C4b deposition assays revealed that T. cruzi activates mainly the lectin and alternative complement pathways in non-immune human serum. Strikingly, we detected that metacyclic trypomastigotes of some T. cruzi strains were highly susceptible to complement-mediated killing in non-immune serum, while other strains were resistant. Furthermore, the rate of parasite invasion in eukaryotic cells was decreased by non-immune serum. Altogether, these results establish that the complement system recognizes T. cruzi metacyclic trypomastigotes, resulting in killing of susceptible strains. The complement system, therefore, acts as a physiological barrier which resistant strains have to evade for successful host infection.

  19. The viability and intestinal epithelial cell adhesion of probiotic strain combination--in vitro study.

    Science.gov (United States)

    Piątek, Jacek; Gibas-Dorna, Magdalena; Olejnik, Anna; Krauss, Hanna; Wierzbicki, Krzysztof; Żukiewicz-Sobczak, Wioletta; Głowacki, Maciej

    2012-01-01

    To be effective, probiotic bacteria must exhibit a number of functional characteristics, including the resistance to gastric acidity and the ability to adhere to the intestinal epithelium. In this study, we examined in vitro the viability of lactic acid bacteria (LAB) combination after exposure to low pH, and the adhesion of LAB to Caco-2 cells during coincubation of 9 bacterial strains. To test bacterial viability, 6 commercially available products were incubated in 0.1 N HCl at pH 1.2 for 60 min. The greatest growth inhibition was noted for the non-capsulated product containing the Lactobacillus rhamnosus strain (log reduction of CFU = 6.4), and the best survival observed for the product containing 9 bacterial strains, equipped with a modern capsule made according to the Multi-Resistant Encapsulation technology (log reduction of CFU = 0.1). In the adhesion experiment, the combination of 9 bacterial strains was added to 17-day-old Caco-2 cell culture for 90 min. The greatest efficiency of adhesion was observed for the inoculum containing 5.5x10(8) CFU/mL/9.6 cm(2) of Caco-2 and the dose of probiotic bacteria of 190 cells per one Caco-2 cell. As a result, approximately 157 bacterial cells adhered to one Caco-2 cell. The results indicate that the combination of 9 bacterial strains in the examined product is characterized as highly adhesive.

  20. Mycobacterium tuberculosis Multidrug Resistant Strain M Induces an Altered Activation of Cytotoxic CD8+ T Cells

    OpenAIRE

    Laura Geffner; Juan Ignacio Basile; Noemí Yokobori; Denise Kviatcovsky; Carmen Sabio y García; Viviana Ritacco; Beatriz López; María del Carmen Sasiain; Silvia de la Barrera

    2014-01-01

    In human tuberculosis (TB), CD8+ T cells contribute to host defense by the release of Th1 cytokines and the direct killing of Mycobacterium tuberculosis (Mtb)-infected macrophages via granule exocytosis pathway or the engagement of receptors on target cells. Previously we demonstrated that strain M, the most prevalent multidrug-resistant (MDR) Mtb strain in Argentine, is a weak inducer of IFN-γ and elicits a remarkably low CD8-dependent cytotoxic T cell activity (CTL). In contrast, the closel...

  1. The SGBS cell strain as a model for the in vitro study of obesity and cancer.

    LENUS (Irish Health Repository)

    Allott, Emma H

    2012-10-01

    The murine adipocyte cell line 3T3-L1 is well characterised and used widely, while the human pre-adipocyte cell strain, Simpson-Golabi-Behmel Syndrome (SGBS), requires validation for use in human studies. Obesity is currently estimated to account for up to 41 % of the worldwide cancer burden. A human in vitro model system is required to elucidate the molecular mechanisms for this poorly understood association. This work investigates the relevance of the SGBS cell strain for obesity and cancer research in humans.

  2. Tnhibitory effect of Fuzheng Yiliuyin in combination with chemotherapeutics on human gastric carcinoma cell strain

    Institute of Scientific and Technical Information of China (English)

    Yi Liu; Rui Wang; Gen-Quan Qiu; Ke-Jun Nan; Xi-Cai Sun

    2006-01-01

    AIM: To study the inhibitory effects of Fuzheng Yiliuyin (Decoction for Suppressing Tumors by Strengthening the Body Resistance) in combination with chemotherapeutics on human gastric carcinoma cell strain.METHODS: Fuzheng Yiliuyin (ZY) combined with various kinds of chemotherapeutics was put into two kinds of cultivated human gastric carcinoma cell strains,then its inhibitory effects on human gastric carcinoma cell strains were determind by the MTT method. Flow cytometer was used to assay the apoptosis rate, and the ultrastructure of gastric carcinoma cells was observed under transmission electron microscope.RESULTS: Obvious apoptosis was seen in gastric carcinoma cells after treatment with ZY for 72 h. ZY and chemical drugs had synergistic inhibition effects on the cultivated gastric carcinoma cells, but the effects were different on various cell strains. The inhibitory effects of ZY could be strengthened by cytotoxic action and apoptosis. ZY combined with fluorouracil, etoposide and cisplatin (EFP) chemotherapeutics had better inhibitory effects on SGC-7901, while ZY combined with EFP or with DDP chemotherapeutics had better inhibitory effects than other drugs on MGC-803.CONCLUSION: ZY induces apoptosis and inhibits the growth of gastric carcinoma cells. ZY has the synergistic function of chemotherapeutics.

  3. 布鲁氏菌M5-90△WboA基因缺失株的构建及免疫效果的初步评价%Construction and preliminary evaluation of immune effects of deletion mutant of WboA gene of Brucella melitensis vaccine M5-90

    Institute of Scientific and Technical Information of China (English)

    张艳; 陈创夫; 张辉; 张沾; 李志强; 张俊波; 孟仁; 王震

    2011-01-01

    Vaccination is a major measure for prevention of brucellosis, but it is unable to be distinguished from the vaccinated to natural infected animals. In this study, the WboA gene was knocked out of the genomic DNA of Brucelh melitensis vaccine M5-90 strain to construct the recombinant M5-90 A WboA by homologous recombination. The test results showed that M5-90 A WboA was less virulent than that of the parent M5-90 strain (p<0.01). Humoral immunity and cellular immunity tests showed that there was no significant difference between M5-90A WboA and M5-90 parent strain (p<0.05). BALB/c mice were immunized with M5-90 A WboA or M5-90 and challenged with virulent strain 16M and the survival rate were 10% and 20%, respectively, indicating that the M5-90 A WboA provided the similar protection of M5-90 strain. Agglutination test and western blot showed that the serum response of M5-90 △ WboA in vaccinated mice were negative. These results indicated that M5-90 A WboA strain might be a promising vaccine against brucellosis, and could be distinguished from the vaccine immunization to natural infection in animals by serum test.%为获得毒力较弱并能区分自然感染和疫苗免疫的布鲁氏菌候选疫苗株,本研究用PCR方法扩增WboA基因的上下游同源臂序列,构建重组质粒pGEM-7zf-△WboA-Sac,电转化布鲁氏菌M5-90感受态细胞,筛选布鲁氏菌疫苗株M5-90的WboA基因缺失株,并对获得的M5-90△WboA遗传稳定性、毒力、免疫保护性、抗体水平等指标进行检测.实验结果表明M5-90△WboA株的毒力比M5-90株明显减弱,差异极显著(p<0.01),体液免疫和细胞免疫结果表明M5-90△WboA株与亲本M5-90株相比差异不显著(p<0.05),M5-90△WboA株和亲本株的保护率分别为10%和20%,表明M5-90△WboA株与M5-90株具有相似的保护性.凝集试验和western blot试验显示M5-90△WboA株免疫小鼠的血清反应结果为阴性.本研究构建的布鲁氏菌基因缺失株M5-90△WboA

  4. The Frequency of Proliferative Stromal Cells in Adipose Tissue Varies Between Inbred Mouse Strains

    Directory of Open Access Journals (Sweden)

    Mo J

    2009-01-01

    Full Text Available Stromal cells derived from adipose tissue (ASCs can proliferate as undifferentiated cells with a fibroblast-like morphology in cell culture, or can be induced to differentiate into a variety of cell types including, adipipogenic, myogenic, neurogenic, osteogenic, chondrogenic and hepatic cells. There is increasing interest to understand the factors controlling the proliferation of ASCs since these cells might provide a readily available source of autologous stem/progenitor cells for cell therapy applications. To explore potential genetic factors that modify the properties of ASCs, we tried to identify relevant properties of ASCs that differ between inbred mouse strains. Plating cells in a modified colony forming assay indicates that the percentage of high proliferative cells among ASCs differs more than 2-fold between 129x1/svj and C57Bl/6J mice. The identification of genetic factors affecting the proliferative capacity of stem cell populations could improve the efficacy of cell therapy.

  5. Entrance and Survival of Brucella pinnipedialis Hooded Seal Strain in Human Macrophages and Epithelial Cells

    Science.gov (United States)

    Briquemont, Benjamin; Sørensen, Karen K.; Godfroid, Jacques

    2013-01-01

    Marine mammal Brucella spp. have been isolated from pinnipeds (B. pinnipedialis) and cetaceans (B. ceti) from around the world. Although the zoonotic potential of marine mammal brucellae is largely unknown, reports of human disease exist. There are few studies of the mechanisms of bacterial intracellular invasion and multiplication involving the marine mammal Brucella spp. We examined the infective capacity of two genetically different B. pinnipedialis strains (reference strain; NTCT 12890 and a hooded seal isolate; B17) by measuring the ability of the bacteria to enter and replicate in cultured phagocytes and epithelial cells. Human macrophage-like cells (THP-1), two murine macrophage cell lines (RAW264.7 and J774A.1), and a human malignant epithelial cell line (HeLa S3) were challenged with bacteria in a gentamicin protection assay. Our results show that B. pinnipedialis is internalized, but is then gradually eliminated during the next 72 – 96 hours. Confocal microscopy revealed that intracellular B. pinnipedialis hooded seal strain colocalized with lysosomal compartments at 1.5 and 24 hours after infection. Intracellular presence of B. pinnipedialis hooded seal strain was verified by transmission electron microscopy. By using a cholesterol-scavenging lipid inhibitor, entrance of B. pinnipedialis hooded seal strain in human macrophages was significantly reduced by 65.8 % (± 17.3), suggesting involvement of lipid-rafts in intracellular entry. Murine macrophages invaded by B. pinnipedialis do not release nitric oxide (NO) and intracellular bacterial presence does not induce cell death. In summary, B. pinnipedialis hooded seal strain can enter human and murine macrophages, as well as human epithelial cells. Intracellular entry of B. pinnipedialis hooded seal strain involves, but seems not to be limited to, lipid-rafts in human macrophages. Brucella pinnipedialis does not multiply or survive for prolonged periods intracellulary. PMID:24376851

  6. Entrance and survival of Brucella pinnipedialis hooded seal strain in human macrophages and epithelial cells.

    Directory of Open Access Journals (Sweden)

    Anett K Larsen

    Full Text Available Marine mammal Brucella spp. have been isolated from pinnipeds (B. pinnipedialis and cetaceans (B. ceti from around the world. Although the zoonotic potential of marine mammal brucellae is largely unknown, reports of human disease exist. There are few studies of the mechanisms of bacterial intracellular invasion and multiplication involving the marine mammal Brucella spp. We examined the infective capacity of two genetically different B. pinnipedialis strains (reference strain; NTCT 12890 and a hooded seal isolate; B17 by measuring the ability of the bacteria to enter and replicate in cultured phagocytes and epithelial cells. Human macrophage-like cells (THP-1, two murine macrophage cell lines (RAW264.7 and J774A.1, and a human malignant epithelial cell line (HeLa S3 were challenged with bacteria in a gentamicin protection assay. Our results show that B. pinnipedialis is internalized, but is then gradually eliminated during the next 72-96 hours. Confocal microscopy revealed that intracellular B. pinnipedialis hooded seal strain colocalized with lysosomal compartments at 1.5 and 24 hours after infection. Intracellular presence of B. pinnipedialis hooded seal strain was verified by transmission electron microscopy. By using a cholesterol-scavenging lipid inhibitor, entrance of B. pinnipedialis hooded seal strain in human macrophages was significantly reduced by 65.8 % (± 17.3, suggesting involvement of lipid-rafts in intracellular entry. Murine macrophages invaded by B. pinnipedialis do not release nitric oxide (NO and intracellular bacterial presence does not induce cell death. In summary, B. pinnipedialis hooded seal strain can enter human and murine macrophages, as well as human epithelial cells. Intracellular entry of B. pinnipedialis hooded seal strain involves, but seems not to be limited to, lipid-rafts in human macrophages. Brucella pinnipedialis does not multiply or survive for prolonged periods intracellulary.

  7. Lattice-strain control of the activity in dealloyed core-shell fuel cell catalysts.

    Science.gov (United States)

    Strasser, Peter; Koh, Shirlaine; Anniyev, Toyli; Greeley, Jeff; More, Karren; Yu, Chengfei; Liu, Zengcai; Kaya, Sarp; Nordlund, Dennis; Ogasawara, Hirohito; Toney, Michael F; Nilsson, Anders

    2010-06-01

    Electrocatalysis will play a key role in future energy conversion and storage technologies, such as water electrolysers, fuel cells and metal-air batteries. Molecular interactions between chemical reactants and the catalytic surface control the activity and efficiency, and hence need to be optimized; however, generalized experimental strategies to do so are scarce. Here we show how lattice strain can be used experimentally to tune the catalytic activity of dealloyed bimetallic nanoparticles for the oxygen-reduction reaction, a key barrier to the application of fuel cells and metal-air batteries. We demonstrate the core-shell structure of the catalyst and clarify the mechanistic origin of its activity. The platinum-rich shell exhibits compressive strain, which results in a shift of the electronic band structure of platinum and weakening chemisorption of oxygenated species. We combine synthesis, measurements and an understanding of strain from theory to generate a reactivity-strain relationship that provides guidelines for tuning electrocatalytic activity.

  8. Generating embryonic stem cells from the inbred mouse strain DBA/2J, a model of glaucoma and other complex diseases.

    Directory of Open Access Journals (Sweden)

    Laura G Reinholdt

    Full Text Available Mouse embryonic stem (ES cells are derived from the inner cell mass of blastocyst stage embryos and are used primarily for the creation of genetically engineered strains through gene targeting. While some inbred strains of mice are permissive to the derivation of embryonic stem cell lines and are therefore easily engineered, others are nonpermissive or recalcitrant. Genetic engineering of recalcitrant strain backgrounds requires gene targeting in a permissive background followed by extensive backcrossing of the engineered allele into the desired strain background. The inbred mouse strain DBA/2J is a recalcitrant strain that is used as a model of many human diseases, including glaucoma, deafness and schizophrenia. Here, we describe the generation of germ-line competent ES cell lines derived from DBA/2J mice. We also demonstrate the utility of DBA/2J ES cells with the creation of conditional knockout allele for Endothelin-2 (Edn2 directly on the DBA/2J strain background.

  9. [Proliferation characteristics of a PK-15 cell-adapted strain of porcine parvovirus].

    Science.gov (United States)

    Wu, Yun-Fei; Zhu, Ling; Xu, Zhi-Wen; Fu, Meng-Jin; Chen, Lei; Yang, Ai-Guo; Guo, Wan-Zhu

    2013-06-01

    To study the proliferation characteristics of PPV in differently infected way and the variance of concentrations in different cells. A strain of porcine parvovirus(PPV) was adapted to PK-15 cells, and a Real-time fluorescent quantitative PCR (FQ-PCR) assay was developed based on the specific region of the NS1 gene of PPV to quantify the PPV. The FQ-PCR was used to measure the viral concentration of virus-infected cells by simultaneous or step by step inoculation and plot one-step growth curves. The proliferation characteristics of PPV strain in different cells lines (HeLa, MDBK, PK-15 ,ST, F81, BHK-21 and Marc-145) was also compared. The results showed the PK-15 cell -adapted strain of PPV produced CPE after 12 passages, and maintained stable CPE at the following 10 messages. The one-step growth curve showed that the virus concentration of simultaneous inoculation was higher than that of the step-by-step inoculation, and the proliferation cycle of step-by-step inoculation was shorter. The proliferation ability of PPV strain in different cells showed that CPE appeared first inPK-15, followed by ST, HeLa and MDBK, and the virus concentration was highest in ST, followed byPK-15, MDBK and HeLa. NO proliferation was observed in F81, BHK-21 and Marc-145 cells. These findings lay a material foundation for the basic researches on PPV and the development of vaccine.

  10. Radioresistant cell strain of human fibrosarcoma cells obtained after long-term exposure to X-rays

    Energy Technology Data Exchange (ETDEWEB)

    Wei, K.; Kodym, R. [Univ. of Vienna (Austria). Department of Radiotherapy and Radiobiology; Jin Cuizheng [Institute of Radiation Medicine, Beijing (China)

    1998-07-01

    A radioresistant cell strain from human fibrosarcoma HT1080 has been obtained after prolonged exposure to x-rays for 7 months (2 Gy per day, 5 days per week). This new strain, HT1080R, differs from HT1080 in a significantly increased ability of clonogenical survival, with coefficient {alpha} decreasing from 0.161 to 0.123 Gy{sup -1} and coefficient {beta} decreasing from 0.0950 to 0.0565 Gy{sup -2}. Furthermore, the radioresistance of HT1080R proved to be stable in long-term passaged cultures as well as in frozen samples. Differences between the two cell lines are also observed in the G-banded karyotype; the new cell line shows monosomy of chromosome 17 and loss of 5p{sup +} and 11q{sup +} present in the parental cells. These data suggest that the radioresistance may have been caused by radiation-induced cell mutation and that the resistant cells may have been selected by repeated irradiations. In order to characterize this new strain, the ability of the cells to rejoin DNA double-strand breaks, the cell cycle distribution and the amount of apoptosis after irradiation have been estimated; however, no differences are observed between these two cell strains. Although the mechanism of the elevated radioresistance remains unknown, this pair of cell strains can provide a new model system for further investigations with regard to the mechanisms of cellular radioresistance. The results also show that any type of irradiation similar to the schedules used in radiotherapy can lead to the formation and selection of more radioresistant cell clones in vitro, a phenomenon with possible implications for radiotherapy. (orig.) With 3 figs., 1 tab., 13 refs.

  11. Plant Cell Wall Degradation by Saprophytic Bacillus subtilis Strains: Gene Clusters Responsible for Rhamnogalacturonan Depolymerization▿

    Science.gov (United States)

    Ochiai, Akihito; Itoh, Takafumi; Kawamata, Akiko; Hashimoto, Wataru; Murata, Kousaku

    2007-01-01

    Plant cell wall degradation is a premier event when Bacillus subtilis, a typical saprophytic bacterium, invades plants. Here we show the degradation system of rhamnogalacturonan type I (RG-I), a component of pectin from the plant cell wall, in B. subtilis strain 168. Strain 168 cells showed a significant growth on plant cell wall polysaccharides such as pectin, polygalacturonan, and RG-I as a carbon source. DNA microarray analysis indicated that three gene clusters (yesOPQRSTUVWXYZ, ytePQRST, and ybcMOPST-ybdABDE) are inducibly expressed in strain 168 cells grown on RG-I. Cells of an industrially important bacterium, B. subtilis strain natto, fermenting soybeans also express the gene cluster including the yes series during the assimilation of soybean used as a carbon source. Among proteins encoded in the yes cluster, YesW and YesX were found to be novel types of RG lyases releasing disaccharide from RG-I. Genetic and enzymatic properties of YesW and YesX suggest that strain 168 cells secrete YesW, which catalyzes the initial cleavage of the RG-I main chain, and the resultant oligosaccharides are converted to disaccharides through the extracellular exotype YesX reaction. The disaccharide is finally degraded into its constituent monosaccharides through the reaction of intracellular unsaturated galacturonyl hydrolases YesR and YteR. This enzymatic route for RG-I degradation in strain 168 differs significantly from that in plant-pathogenic fungus Aspergillus aculeatus. This is, to our knowledge, the first report on the bacterial system for complete RG-I main chain degradation. PMID:17449691

  12. Plant cell wall degradation by saprophytic Bacillus subtilis strains: gene clusters responsible for rhamnogalacturonan depolymerization.

    Science.gov (United States)

    Ochiai, Akihito; Itoh, Takafumi; Kawamata, Akiko; Hashimoto, Wataru; Murata, Kousaku

    2007-06-01

    Plant cell wall degradation is a premier event when Bacillus subtilis, a typical saprophytic bacterium, invades plants. Here we show the degradation system of rhamnogalacturonan type I (RG-I), a component of pectin from the plant cell wall, in B. subtilis strain 168. Strain 168 cells showed a significant growth on plant cell wall polysaccharides such as pectin, polygalacturonan, and RG-I as a carbon source. DNA microarray analysis indicated that three gene clusters (yesOPQRSTUVWXYZ, ytePQRST, and ybcMOPST-ybdABDE) are inducibly expressed in strain 168 cells grown on RG-I. Cells of an industrially important bacterium, B. subtilis strain natto, fermenting soybeans also express the gene cluster including the yes series during the assimilation of soybean used as a carbon source. Among proteins encoded in the yes cluster, YesW and YesX were found to be novel types of RG lyases releasing disaccharide from RG-I. Genetic and enzymatic properties of YesW and YesX suggest that strain 168 cells secrete YesW, which catalyzes the initial cleavage of the RG-I main chain, and the resultant oligosaccharides are converted to disaccharides through the extracellular exotype YesX reaction. The disaccharide is finally degraded into its constituent monosaccharides through the reaction of intracellular unsaturated galacturonyl hydrolases YesR and YteR. This enzymatic route for RG-I degradation in strain 168 differs significantly from that in plant-pathogenic fungus Aspergillus aculeatus. This is, to our knowledge, the first report on the bacterial system for complete RG-I main chain degradation.

  13. Predicting cell viability within tissue scaffolds under equiaxial strain: multi-scale finite element model of collagen-cardiomyocytes constructs.

    Science.gov (United States)

    Elsaadany, Mostafa; Yan, Karen Chang; Yildirim-Ayan, Eda

    2017-01-16

    Successful tissue engineering and regenerative therapy necessitate having extensive knowledge about mechanical milieu in engineered tissues and the resident cells. In this study, we have merged two powerful analysis tools, namely finite element analysis and stochastic analysis, to understand the mechanical strain within the tissue scaffold and residing cells and to predict the cell viability upon applying mechanical strains. A continuum-based multi-length scale finite element model (FEM) was created to simulate the physiologically relevant equiaxial strain exposure on cell-embedded tissue scaffold and to calculate strain transferred to the tissue scaffold (macro-scale) and residing cells (micro-scale) upon various equiaxial strains. The data from FEM were used to predict cell viability under various equiaxial strain magnitudes using stochastic damage criterion analysis. The model validation was conducted through mechanically straining the cardiomyocyte-encapsulated collagen constructs using a custom-built mechanical loading platform (EQUicycler). FEM quantified the strain gradients over the radial and longitudinal direction of the scaffolds and the cells residing in different areas of interest. With the use of the experimental viability data, stochastic damage criterion, and the average cellular strains obtained from multi-length scale models, cellular viability was predicted and successfully validated. This methodology can provide a great tool to characterize the mechanical stimulation of bioreactors used in tissue engineering applications in providing quantification of mechanical strain and predicting cellular viability variations due to applied mechanical strain.

  14. Strain-balanced MQW pin solar cells grown using a robot-loading showerhead reactor

    Science.gov (United States)

    Roberts, J. S.; Airey, R.; Hill, G.; Calder, C.; Barnham, K. W. J.; Lynch, M.; Tibbits, T.; Johnson, D.; Pakes, A.; Grantham, T.

    2007-01-01

    A touch-screen controlled, robot-loading system for the Thomas Swan 7×2 flip-top showerhead reactor has been developed. The reactor has been configured for the growth of GaAs and InP materials and has been used to prepare strain-balanced MQW (SBMQW) pin solar cell material on GaAs substrates. Both material characterisation and solar cell performance for SBMQW pin cells are described.

  15. Lattice-Strain Control of Exceptional Activity in Dealloyed Core-Shell Fuel Cell Catalysts

    Energy Technology Data Exchange (ETDEWEB)

    Strasser, Peter

    2011-08-19

    We present a combined experimental and theoretical approach to demonstrate how lattice strain can be used to continuously tune the catalytic activity of the oxygen reduction reaction (ORR) on bimetallic nanoparticles that have been dealloyed. The sluggish kinetics of the ORR is a key barrier to the adaptation of fuel cells and currently limits their widespread use. Dealloyed Pt-Cu bimetallic nanoparticles, however, have been shown to exhibit uniquely high reactivity for this reaction. We first present evidence for the formation of a core-shell structure during dealloying, which involves removal of Cu from the surface and subsurface of the precursor nanoparticles. We then show that the resulting Pt-rich surface shell exhibits compressive strain that depends on the composition of the precursor alloy. We next demonstrate the existence of a downward shift of the Pt d-band, resulting in weakening of the bond strength of intermediate oxygenated species due to strain. Finally, we combine synthesis, strain, and catalytic reactivity in an experimental/theoretical reactivity-strain relationship which provides guidelines for the rational design of strained oxygen reduction electrocatalysts. The stoichiometry of the precursor, together with the dealloying conditions, provides experimental control over the resulting surface strain and thereby allows continuous tuning of the surface electrocatalytic reactivity - a concept that can be generalized to other catalytic reactions.

  16. Cell culture and animal infection with distinct Trypanosoma cruzi strains expressing red and green fluorescent proteins.

    Science.gov (United States)

    Pires, S F; DaRocha, W D; Freitas, J M; Oliveira, L A; Kitten, G T; Machado, C R; Pena, S D J; Chiari, E; Macedo, A M; Teixeira, S M R

    2008-03-01

    Different strains of Trypanosoma cruzi were transfected with an expression vector that allows the integration of green fluorescent protein (GFP) and red fluorescent protein (RFP) genes into the beta-tubulin locus by homologous recombination. The sites of integration of the GFP and RFP markers were determined by pulse-field gel electrophoresis and Southern blot analyses. Cloned cell lines selected from transfected epimastigote populations maintained high levels of fluorescent protein expression even after 6 months of in vitro culture of epimastigotes in the absence of drug selection. Fluorescent trypomastigotes and amastigotes were observed within Vero cells in culture as well as in hearts and diaphragms of infected mice. The infectivity of the GFP- and RFP-expressing parasites in tissue culture cells was comparable to wild type populations. Furthermore, GFP- and RFP-expressing parasites were able to produce similar levels of parasitemia in mice compared with wild type parasites. Cell cultures infected simultaneously with two cloned cell lines from the same parasite strain, each one expressing a distinct fluorescent marker, showed that at least two different parasites are able to infect the same cell. Double-infected cells were also detected when GFP- and RFP-expressing parasites were derived from strains belonging to two distinct T. cruzi lineages. These results show the usefulness of parasites expressing GFP and RFP for the study of various aspects of T. cruzi infection including the mechanisms of cell invasion, genetic exchange among parasites and the differential tissue distribution in animal models of Chagas disease.

  17. Cyclic strain alters the expression and release of angiogenic factors by human tendon cells.

    Science.gov (United States)

    Mousavizadeh, Rouhollah; Khosravi, Shahram; Behzad, Hayedeh; McCormack, Robert G; Duronio, Vincent; Scott, Alex

    2014-01-01

    Angiogenesis is associated with the tissue changes underlying chronic overuse tendinopathy. We hypothesized that repetitive, cyclic loading of human tendon cells would lead to increased expression and activity of angiogenic factors. We subjected isolated human tendon cells to overuse tensile loading using an in vitro model (1 Hz, 10% equibiaxial strain). We found that mechanically stimulated human tendon cells released factors that promoted in vitro proliferation and tube formation by human umbilical vein endothelial cells (HUVEC). In response to cyclic strain, there was a transient increase in the expression of several angiogenic genes including ANGPTL4, FGF-2, COX-2, SPHK1, TGF-alpha, VEGF-A and VEGF-C, with no change in anti-angiogenic genes (BAI1, SERPINF1, THBS1 and 2, TIMP1-3). Cyclic strain also resulted in the extracellular release of ANGPTL4 protein by tendon cells. Our study is the first report demonstrating the induction of ANGPTL4 mRNA and release of ANGPTL4 protein in response to cyclic strain. Tenocytes may contribute to the upregulation of angiogenesis during the development of overuse tendinopathy.

  18. In vitro permissivity of bovine cells for wild-type and vaccinal myxoma virus strains

    Directory of Open Access Journals (Sweden)

    Foucras Gilles

    2007-09-01

    Full Text Available Abstract Myxoma virus (MYXV, a leporide-specific poxvirus, represents an attractive candidate for the generation of safe, non-replicative vaccine vector for non-host species. However, there is very little information concerning infection of non-laboratory animals species cells with MYXV. In this study, we investigated interactions between bovine cells and respectively a wild type strain (T1 and a vaccinal strain (SG33 of MYXV. We showed that bovine KOP-R, BT and MDBK cell lines do not support MYXV production. Electron microscopy observations of BT-infected cells revealed the low efficiency of viral entry and the production of defective virions. In addition, infection of bovine peripheral blood mononuclear cells (PBMC occurred at a very low level, even following non-specific activation, and was always abortive. We did not observe significant differences between the wild type strain and the vaccinal strain of MYXV, indicating that SG33 could be used for new bovine vaccination strategies.

  19. Effects of navelbine and docetaxel on gene expression in lung cancer cell strains

    Institute of Scientific and Technical Information of China (English)

    Li CAI; Hai-ying DONG; Guang-jie SUI

    2005-01-01

    Aim: To search genes sensitivity to the anti-cancer drugs navelbine (NVB) and docetaxel (DOC) in small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC) cell strains. Methods: The sensitivity of 4 strains of SCLC and 6 strains of NSCLC to NVB and DOC was evaluated using the MTT assay. The expression of 1291 sensitive-related genes to the anti-cancer drugs in 10 lung cancer cell strains was measured using cDNA macroarrays and the relationship was analyzed.Results: In total, there were 56 (r≥0.4) genes sensitive to NVB and DOC. For NVB: 36 genes were sensitive to NVB, 20 co-expressed genes between the SCLC+NSCLC set and the NSCLC set; 27 expressed genes and 7 specially expressed genes in the SCLC+NSCLC set; and 29 expressed genes and 9 specially expressed genes in the NSCLC set. For DOC, 50 genes were sensitive to DOC, 12co-expressed genes between the SCLC+NSCLC set and the NSCLC set; 24expressed genes and 12 specially expressed genes in the SCLC+NSCLC set; and 38 expressed genes and 26 specially expressed genes in the NSCLC set. The genes sensitive to NVB and DOC in lung-cancer cell stains were mainly divided into the following 4 categories: signal transduction molecules, cell factors, transcription factors and metabolism-related enzymes and inhibitors. Conclusions:There were obvious differences in genes related to NVB and DOC between SCLC and NSCLC cell strains, but the same as categories of function.

  20. Three-dimensional development of tensile pre-strained annulus fibrosus cells for tissue regeneration: An in-vitro study

    Energy Technology Data Exchange (ETDEWEB)

    Chuah, Yon Jin [School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore 637459 (Singapore); Lee, Wu Chean [University Hospital Conventry & Warwickshire NHS Trust, Clifford Bridge Road, West Midlands CV2, 2DX (United Kingdom); Wong, Hee Kit [Department of Orthopedic Surgery, National University Health System, NUHS Tower Block Level 11, 1E Kent Ridge Road, Singapore 119228 (Singapore); Kang, Yuejun, E-mail: yuejun.kang@ntu.edu.sg [School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore 637459 (Singapore); Hee, Hwan Tak, E-mail: HTHee@ntu.edu.sg [School of Chemical and Biomedical Engineering, Nanyang Technological University, 62 Nanyang Drive, Singapore 637459 (Singapore); Pinnacle Spine & Scoliosis Centre, 3 Mount Elizabeth, Mount Elizabeth Medical Centre, #04-07, Singapore 228510 (Singapore); School of Physical and Mathematical Sciences, Nanyang Technological University, 21 Nanyang Link, Singapore 637459 (Singapore)

    2015-02-01

    Prior research has investigated the immediate response after application of tensile strain on annulus fibrosus (AF) cells for the past decade. Although mechanical strain can produce either catabolic or anabolic consequences to the cell monolayer, little is known on how to translate these findings into further tissue engineering applications. Till to date, the application and effect of tensile pre-strained cells to construct a three-dimensional (3D) AF tissue remains unknown. This study aims to investigate the effect of tensile pre-strained exposure of 1 to 24 h on the development of AF pellet culture for 3 weeks. Equibiaxial cyclic tensile strain was applied on AF monolayer cells over a period of 24 h, which was subsequently developed into a cell pellet. Investigation on cellular proliferation, phenotypic gene expression, and histological changes revealed that tensile pre-strain for 24 h had significant and lasting effect on the AF tissue development, with enhanced cell proliferation, and up-regulation of collagen type I, II, and aggrecan expression. Our results demonstrated the regenerative ability of AF cell pellets subjected to 24 h tensile pre-straining. Knowledge on the effects of tensile pre-strain exposure is necessary to optimize AF development for tissue reconstruction. Moreover, the tensile pre-strained cells may further be utilized in either cell therapy to treat mild disc degeneration disease, or the development of a disc construct for total disc replacement. - Highlights: • Establishment of tensile pre-strained cell line population for annulus development. • Tensile strain limits collagen gene expression declination in monolayer culture. • Tensile pre-strained cells up-regulate their matrix protein in 3D pellet culture.

  1. Airway smooth muscle cell tone amplifies contractile function in the presence of chronic cyclic strain.

    Science.gov (United States)

    Fairbank, Nigel J; Connolly, Sarah C; Mackinnon, James D; Wehry, Kathrin; Deng, Linhong; Maksym, Geoffrey N

    2008-09-01

    Chronic contractile activation, or tone, in asthma coupled with continuous stretching due to breathing may be involved in altering the contractile function of airway smooth muscle (ASM). Previously, we (11) showed that cytoskeletal remodeling and stiffening responses to acute (2 h) localized stresses were modulated by the level of contractile activation of ASM. Here, we investigated if altered contractility in response to chronic mechanical strain was dependent on repeated modulation of contractile tone. Cultured human ASM cells received 5% cyclic (0.3 Hz), predominantly uniaxial strain for 5 days, with once-daily dosing of either sham, forskolin, carbachol, or histamine to alter tone. Stiffness, contractility (KCl), and "relaxability" (forskolin) were then measured as was cell alignment, myosin light-chain phosphorylation (pMLC), and myosin light-chain kinase (MLCK) content. Cells became aligned and baseline stiffness increased with strain, but repeated lowering of tone inhibited both effects (P negative tone-modulation dependence of MLCK, observed in static conditions in agreement with previous reports, with strain and tone together increasing both MLCK and pMLC. Furthermore, contractility increased 176% (SE 59) with repeated tone elevation. These findings indicate that with strain, and not without, repeated tone elevation promoted contractile function through changes in cytoskeletal organization and increased contractile protein. The ability of repeated contractile activation to increase contractility, but only with mechanical stretching, suggests a novel mechanism for increased ASM contractility in asthma and for the role of continuous bronchodilator and corticosteroid therapy in reversing airway hyperresponsiveness.

  2. A biomechanical model for fluidization of cells under dynamic strain.

    Science.gov (United States)

    Wu, Tenghu; Feng, James J

    2015-01-06

    Recent experiments have investigated the response of smooth muscle cells to transient stretch-compress (SC) and compress-stretch (CS) maneuvers. The results indicate that the transient SC maneuver causes a sudden fluidization of the cell while the CS maneuver does not. To understand this asymmetric behavior, we have built a biomechanical model to probe the response of stress fibers to the two maneuvers. The model couples the cross-bridge cycle of myosin motors with a viscoelastic Kelvin-Voigt element that represents the stress fiber. Simulation results point to the sensitivity of the myosin detachment rate to tension as the cause for the asymmetric response of the stress fiber to the CS and SC maneuvers. For the SC maneuver, the initial stretch increases the tension in the stress fiber and suppresses myosin detachment. The subsequent compression then causes a large proportion of the myosin population to disengage rapidly from actin filaments. This leads to the disassembly of the stress fibers and the observed fluidization. In contrast, the CS maneuver only produces a mild loss of myosin motors and no fluidization.

  3. Cell culture isolation and sequence analysis of genetically diverse US porcine epidemic diarrhea virus strains including a novel strain with a large deletion in the spike gene.

    Science.gov (United States)

    Oka, Tomoichiro; Saif, Linda J; Marthaler, Douglas; Esseili, Malak A; Meulia, Tea; Lin, Chun-Ming; Vlasova, Anastasia N; Jung, Kwonil; Zhang, Yan; Wang, Qiuhong

    2014-10-10

    The highly contagious and deadly porcine epidemic diarrhea virus (PEDV) first appeared in the US in April 2013. Since then the virus has spread rapidly nationwide and to Canada and Mexico causing high mortality among nursing piglets and significant economic losses. Currently there are no efficacious preventive measures or therapeutic tools to control PEDV in the US. The isolation of PEDV in cell culture is the first step toward the development of an attenuated vaccine, to study the biology of PEDV and to develop in vitro PEDV immunoassays, inactivation assays and screen for PEDV antivirals. In this study, nine of 88 US PEDV strains were isolated successfully on Vero cells with supplemental trypsin and subjected to genomic sequence analysis. They differed genetically mainly in the N-terminal S protein region as follows: (1) strains (n=7) similar to the highly virulent US PEDV strains; (2) one similar to the reportedly US S INDEL PEDV strain; and (3) one novel strain most closely related to highly virulent US PEDV strains, but with a large (197aa) deletion in the S protein. Representative strains of these three genetic groups were passaged serially and grew to titers of ∼5-6log10 plaque forming units/mL. To our knowledge, this is the first report of the isolation in cell culture of an S INDEL PEDV strain and a PEDV strain with a large (197aa) deletion in the S protein. We also designed primer sets to detect these genetically diverse US PEDV strains. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Biodegradation of pesticide profenofos by the free and immobilized cells of Pseudoxanthomonas suwonensis strain HNM.

    Science.gov (United States)

    Talwar, Manjunatha P; Ninnekar, Harichandra Z

    2015-09-01

    Profenofos is an organophosphate pesticide used extensively in agriculture to control pests. A bacterium capable of degrading profenofos was isolated from pesticide-contaminated soil samples and identified as Pseudoxanthomonas suwonensis strain HNM based on its morphological and biochemical characteristics and phylogenetic analysis of 16S rRNA gene sequences. 4-Bromo-2-chlorophenol was identified as a metabolite of profenofos degradation by HPLC and GC-MS analysis. The organism degraded profenofos by hydrolysis to yield 4-bromo-2-chlorophenol which was further utilized as carbon source for growth. The organism utilized various organophosphate pesticides such as temephos, quinalphos, and chloropyrifos as carbon sources. The optimum conditions for degradation of profenofos by P. suwonensis strain HMN were found to be at pH 7 and 30 °C. We have investigated the rate of degradation of profenofos by the free and immobilized cells of P. suwonensis strain HNM in various matrices such as sodium alginate (SA), sodium alginate-polyvinyl alcohol (SA-PVA), and SA-bentonite clay. The rate of degradation of 3 and 6 mM profenofos by the freely suspended cells were compared with that by immobilized cells in batches and semi-continuous with shaken cultures. The SA-bentonite clay-immobilized cells showed higher rate of degradation of 3 and 6 mM profenofos then freely suspended cells and cells immobilized in SA and SA-PVA. The SA-bentonite clay-immobilized cells of P. suwonensis strain HNM could be reused for more than 32 cycles without losing their degradation capacity. Thus, the immobilized cells are more efficient than freely suspended cells for the degradation of organophosphate pesticide contaminated water.

  5. iPSC-derived human mesenchymal stem cells improve myocardial strain of infarcted myocardium.

    Science.gov (United States)

    Miao, Qingfeng; Shim, Winston; Tee, Nicole; Lim, Sze Yun; Chung, Ying Ying; Ja, K P Myu Mia; Ooi, Ting Huay; Tan, Grace; Kong, Geraldine; Wei, Heming; Lim, Chong Hee; Sin, Yoong Kong; Wong, Philip

    2014-08-01

    We investigated global and regional effects of myocardial transplantation of human induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs) in infarcted myocardium. Acute myocardial infarction (MI) was induced by ligation of left coronary artery of severe combined immunodeficient mice before 2 × 10(5) iMSCs or cell-free saline were injected into peri-infarcted anterior free wall. Sham-operated animals received no injection. Global and regional myocardial function was assessed serially at 1-week and 8-week by segmental strain analysis by using two dimensional (2D) speckle tracking echocardiography. Early myocardial remodelling was observed at 1-week and persisted to 8-week with global contractility of ejection fraction and fractional area change in saline- (32.96 ± 14.23%; 21.50 ± 10.07%) and iMSC-injected (32.95 ± 10.31%; 21.00 ± 7.11%) groups significantly depressed as compared to sham control (51.17 ± 11.69%, P myocardial dilatation was observed in saline-injected animals (4.40 ± 0.62 mm, P strain analysis showed significant improved basal anterior wall strain (28.86 ± 8.16%, P strain only in saline-injected (21.50 ± 5.31%, P myocardial strain coincided with the presence of interconnecting telocytes in interstitial space of the infarcted anterior segment of the heart. Our results show that localized injection of iMSCs alleviates ventricular remodelling, sustains global and regional myocardial strain by paracrine-driven effect on neoangiogenesis and myocardial deformation/compliance via parenchymal and interstitial cell interactions in the infarcted myocardium.

  6. Helicobacter pylori strains vary cell shape and flagellum number to maintain robust motility in viscous environments.

    Science.gov (United States)

    Martínez, Laura E; Hardcastle, Joseph M; Wang, Jeffrey; Pincus, Zachary; Tsang, Jennifer; Hoover, Timothy R; Bansil, Rama; Salama, Nina R

    2016-01-01

    The helical shape of the human stomach pathogen Helicobacter pylori has been suggested to provide mechanical advantage for penetrating the viscous stomach mucus layer. Using single-cell tracking and quantitative morphology analysis, we document marked variation in cell body helical parameters and flagellum number among H. pylori strains leading to distinct and broad speed distributions in broth and viscous gastric mucin media. These distributions reflect both temporal variation in swimming speed and morphologic variation within the population. Isogenic mutants with straight-rod morphology showed 7-21% reduction in speed and a lower fraction of motile bacteria. Mutational perturbation of flagellum number revealed a 19% increase in speed with 4 versus 3 median flagellum number. Resistive force theory modeling incorporating variation of both cell shape and flagellum number predicts qualitative speed differences of 10-30% among strains. However, quantitative comparisons suggest resistive force theory underestimates the influence of cell body shape on speed for helical shaped bacteria.

  7. The microstructural origin of strain hardening in two-dimensional open-cell metal foams

    NARCIS (Netherlands)

    Mangipudi, K. R.; van Buuren, S. W.; Onck, P. R.

    2010-01-01

    This paper aims at elucidating the microstructural origin of strain hardening in open-cell metal foams. We have developed a multiscale model that allows to study the development of plasticity at two length scales: (i) the development of plastic zones inside individual struts (microscopic scale) and

  8. Pathogenic potential of Escherichia coli clinical strains from orthopedic implant infections towards human osteoblastic cells

    Science.gov (United States)

    Crémet, Lise; Broquet, Alexis; Brulin, Bénédicte; Jacqueline, Cédric; Dauvergne, Sandie; Brion, Régis; Asehnoune, Karim; Corvec, Stéphane; Heymann, Dominique; Caroff, Nathalie

    2015-01-01

    Escherichia coli is one of the first causes of Gram-negative orthopedic implant infections (OII), but little is known about the pathogenicity of this species in such infections that are increasing due to the ageing of the population. We report how this pathogen interacts with human osteoblastic MG-63 cells in vitro, by comparing 20 OII E. coli strains to two Staphylococcus aureus and two Pseudomonas aeruginosa strains. LDH release assay revealed that 6/20 (30%) OII E. coli induced MG-63 cell lysis whereas none of the four control strains was cytotoxic after 4 h of coculture. This high cytotoxicity was associated with hemolytic properties and linked to hlyA gene expression. We further showed by gentamicin protection assay and confocal microscopy that the non-cytotoxic E. coli were not able to invade MG-63 cells unlike S. aureus strains (internalization rate coli versus 8.88 ± 2.31% and 4.60 ± 0.42% for both S. aureus). The non-cytotoxic E. coli also demonstrated low adherence rates (coli eliciting higher IL-6 and TNF-α mRNA expression in the osteoblastic cells. Either highly cytotoxic or slightly invasive OII E. coli do not show the same infection strategies as S. aureus towards osteoblasts. PMID:26333570

  9. Quasi-monolithic planar load cells using built-in resonant strain gauges

    NARCIS (Netherlands)

    Tilmans, Harrie A.C.; Elwenspoek, Miko

    1993-01-01

    Two load cell designs are presented using resonant strain gauges providing a frequency output. One design is based on a four-point beam deflection jig. It offers high sensitivity, but suffers from robustness and impractical geometries for a broad force range. A modified planar design (typical dimens

  10. Enhanced caffeine degradation by immobilised cells of Leifsonia sp. strain SIU.

    Science.gov (United States)

    Ibrahim, Salihu; Shukor, Mohd Y; Syed, Mohd A; Johari, Wan L W; Shamaan, Nor A; Sabullah, Mohd K; Ahmad, Siti A

    2016-01-01

    In a previous study, we isolated Leifsonia sp. strain SIU, a new bacterium from agricultured soil. The bacterium was tested for its ability to degrade caffeine. The isolate was encapsulated in gellan gum and its ability to degrade caffeine was compared with the free cells. The optimal caffeine degradation was attained at a gellan gum concentration of 0.75% (w/v), a bead size of 4 mm diameter, and 250 beads per 100 mL of medium. At a caffeine concentration of 0.1 g/L, immobilised cells of the strain SIU degraded caffeine within 9 h, which is faster when compared to the case of free cells, in which it took 12 h to degrade. The immobilised cells degraded caffeine completely within 39 and 78 h at 0.5 and 1.0 g/L, while the free cells took 72 and 148 h at 0.5 and 1.0 g/L, respectively. At higher caffeine concentrations, immobilised cells exhibited a higher caffeine degradation rate. At concentrations of 1.5 and 2.0 g/L, caffeine-degrading activities of both immobilised and free cells were inhibited. The immobilised cells showed no loss in caffeine-degrading activity after being used repeatedly for nine 24-h cycles. The effect of heavy metals on immobilised cells was also tested. This study showed an increase in caffeine degradation efficiency when the cells were encapsulated in gellan gum.

  11. Effects of Saponins against Clinical E. coli Strains and Eukaryotic Cell Line

    Directory of Open Access Journals (Sweden)

    Michał Arabski

    2012-01-01

    Full Text Available Saponins are detergent-like substances showing antibacterial as well as anticancer potential. In this study, the effects of saponins from Quillaja saponaria were analyzed against prokaryotic and eukaryotic cells. Multidrug-resistant clinical E. coli strains were isolated from human urine. As eukaryotic cells, the CHO-K1 cell lines were applied. Antibacterial effect of ampicillin, streptomycin, and ciprofloxacin in the presence of saponins was measured by cultivation methods. Properties of saponins against CHO-K1 cells were measured by the MTT test, hemolysis assay and flow cytometry. Saponin from Quillaja saponaria has a cytotoxic effect at concentrations higher than 25 μg/mL and in the range of 12–50 μg/mL significantly increases the level of early apoptotic cells. Saponin at dose of 12 μg/mL enhances the six E. coli strains growth. We postulate that saponins increase the influx of nutrients from the medium into E. coli cells. Saponins do not have synergetic effects on antibacterial action of tested antibiotics. In contrary, in the presence of saponins and antibiotics, more CFU/mL E. coli cells were observed. This effect was similar to saponins action alone towards E. coli cells. In conclusion, saponins was cytotoxic against CHO-K1 cells, whereas against E. coli cells this effect was not observed.

  12. VUV treatment combined with mechanical strain of stretchable polymer foils resulting in cell alignment

    Energy Technology Data Exchange (ETDEWEB)

    Barb, R.-A. [Institute of Applied Physics, Johannes Kepler University Linz (Austria); Magnus, B. [Innovacell Biotechnologie AG, Innsbruck (Austria); Innerbichler, S. [Innerbichler GmbH, Breitenbach am Inn (Austria); Greunz, T. [CDL-MS-MACH, Johannes Kepler University Linz (Austria); Wiesbauer, M. [Institute of Applied Physics, Johannes Kepler University Linz (Austria); Marksteiner, R. [Innovacell Biotechnologie AG, Innsbruck (Austria); Stifter, D. [CDL-MS-MACH, Johannes Kepler University Linz (Austria); Heitz, J., E-mail: johannes.heitz@jku.at [Institute of Applied Physics, Johannes Kepler University Linz (Austria)

    2015-01-15

    Highlights: • Elastic polyurethane (PU) foils were exposed to the vacuum-UV in reactive atmosphere. • The photomodification resulted in improved cytocompatibilty. • Parallel microgrooves formed on the irradiated PU surfaces after strong elongation. • Cells seeded onto microgrooves aligned their shapes in the direction of the grooves. • Elongation occurred also for cells on PU subjected to cyclic mechanical stretching. - Abstract: Cell-alignment along a defined direction can have a direct effect on the cell functionality and differentiation. Oriented micro- or nanotopographic structures on cell culture substrates can induce cell-alignment. Surface chemistry, wettability, and stiffness of the substrate are also important material features as they strongly influence the cell–substrate interactions. For improved bio-compatibility, highly elastic polyurethane (PU) foils were exposed to the vacuum-UV (VUV) light of a Xe{sub 2}{sup *} excimer lamp at 172 nm in a nitrogen containing atmosphere (N{sub 2} or NH{sub 3}). The irradiation resulted in a change in the chemical surface composition. Additionally, the formation of regular parallel microgrooves was observed on the irradiated surfaces after strong uni-axial deformation (i.e., more than about 50% strain) of the photo-modified PU foils. Cell seeding experiments demonstrated that the VUV modified polymer foils strongly enhance cell adhesion and proliferation. Cells seeded onto microgrooves aligned their shapes and elongated in the direction of the grooves. A similar effect was observed for cells seeded on photo-modified PU foils subjected to cyclic mechanical stretching at lower strain levels (i.e., typically 10% strain) without groove-formation. The cells had also here an elongated shape, however they not always align in a defined direction relative to the stretching.

  13. Mechanism of Cell Culture Adaptation of an Enteric Calicivirus, the Porcine Sapovirus Cowden Strain.

    Science.gov (United States)

    Lu, Zhongyan; Yokoyama, Masaru; Chen, Ning; Oka, Tomoichiro; Jung, Kwonil; Chang, Kyeong-Ok; Annamalai, Thavamathi; Wang, Qiuhong; Saif, Linda J

    2015-11-18

    The porcine sapovirus (SaV) (PoSaV) Cowden strain is one of only a few culturable enteric caliciviruses. Compared to the wild-type (WT) PoSaV Cowden strain, tissue culture-adapted (TC) PoSaV has two conserved amino acid substitutions in the RNA-dependent RNA polymerase (RdRp) and six in the capsid protein (VP1). By using the reverse-genetics system, we identified that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328), but not the substitutions in the RdRp region, were critical for the cell culture adaptation of the PoSaV Cowden strain. The other two substitutions in VP1 (residues 291 and 295) reduced virus replication in vitro. Three-dimensional (3D) structural analysis of VP1 showed that residue 178 was located near the dimer-dimer interface, which may affect VP1 assembly and oligomerization; residues 289, 291, 324, and 328 were located at protruding subdomain 2 (P2) of VP1, which may influence virus binding to cellular receptors; and residue 295 was located at the interface of two monomeric VP1 proteins, which may influence VP1 dimerization. Although reversion of the mutation at residue 291 or 295 from that of the TC strain to that of the WT reduced virus replication in vitro, it enhanced virus replication in vivo, and the revertants induced higher-level serum and mucosal antibody responses than those induced by the TC PoSaV Cowden strain. Our findings reveal the molecular basis for PoSaV adaptation to cell culture. These findings may provide new, critical information for the cell culture adaptation of other PoSaV strains and human SaVs or noroviruses. The tissue culture-adapted porcine sapovirus Cowden strain is one of only a few culturable enteric caliciviruses. We discovered that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328) were critical for its adaptation to LLC-PK cells. Two substitutions in VP1 (residues 291 and 295) reduced virus replication in vitro but enhanced virus replication and induced higher-level serum and

  14. Exposure to Varying Strain Magnitudes Influences the Conversion of Normal Skin Fibroblasts Into Hypertrophic Scar Cells.

    Science.gov (United States)

    Kuang, Ruixia; Wang, Zhiguo; Xu, Quanchen; Cai, Xia; Liu, Tao

    2016-04-01

    Mechanical strain is a key contributor in the pathogenesis of hypertrophic scarring, whose optimal stretch magnitudes to initiate the differentiation of normal skin fibroblasts into aberrant fibroblasts phenotype remains largely unresolved. Influence of varying cyclic strain magnitudes on cultured human normal skin fibroblasts and its transformation into hypertrophic scar fibroblast-like phenotype is investigated in this study. Cultured fibroblasts isolated from hypertrophic scar and normal skin tissue were subjected to cyclic mechanical stretching under individual 10%, 15%, and 20% strain magnitudes at a frequency of 0.1 Hz for 24 hours. Stretched normal skin fibroblasts demonstrated significantly increased rates of cell proliferation, and also apparently oriented away nearly perpendicular to the applied stretching direction. Interestingly, the applied 10% strains magnitude resulted in a markedly enhanced cell proliferative ability compared with that of 20% strain magnitude. Parameters involving the mechanotransduction signaling, such as integrin β1 and P130Cas, were significantly improved at both mRNA and protein levels in the stretched normal skin fibroblasts, which was demonstrated in a negative magnitude-dependent manner. In addition, 10% strains magnitude triggered the highest expression levels of growth factor TGF-β1 and collagen matrix in stretched normal skin fibroblasts. Collectively, these results indicate that the 10% stretching magnitude, of the 3 strain magnitudes studied, is most effective for triggering the optimal mechanotransduction effects and biological responses inside cultured skin fibroblasts. The demonstrable conversion of normal skin fibroblasts into hypertrophic scar fibroblasts was also observed when 10% stretching magnitude was applied to cultured fibroblasts in vitro.

  15. Biodegradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1

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    Preeti N. Tallur

    2015-09-01

    Full Text Available Pyrethroid pesticide cypermethrin is a environmental pollutant because of its widespread use, toxicity and persistence. Biodegradation of such chemicals by microorganisms may provide an cost-effective method for their detoxification. We have investigated the degradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1 in various matrices such as, polyurethane foam (PUF, polyacrylamide, sodium alginate and agar. The optimum temperature and pH for the degradation of cypermethrin by immobilized cells of Micrococcus sp. were found to be 30 °C and 7.0, respectively. The rate of degradation of 10 and 20 mM of cypermethrin by freely suspended cells were compared with that of immobilized cells in batches and semi-continuous with shaken cultures. PUF-immobilized cells showed higher degradation of cypermethrin (10 mM and 20 mM than freely suspended cells and cells immobilized in other matrices. The PUF-immobilized cells of Micrococcus sp. strain CPN 1 were retain their degradation capacity. Thus, they can be reused for more than 32 cycles, without losing their degradation capacity. Hence, the PUF-immobilized cells of Micrococcus sp. could potentially be used in the bioremediation of cypermethrin contaminated water.

  16. Biodegradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1.

    Science.gov (United States)

    Tallur, Preeti N; Mulla, Sikandar I; Megadi, Veena B; Talwar, Manjunatha P; Ninnekar, Harichandra Z

    2015-01-01

    Pyrethroid pesticide cypermethrin is a environmental pollutant because of its widespread use, toxicity and persistence. Biodegradation of such chemicals by microorganisms may provide an cost-effective method for their detoxification. We have investigated the degradation of cypermethrin by immobilized cells of Micrococcus sp. strain CPN 1 in various matrices such as, polyurethane foam (PUF), polyacrylamide, sodium alginate and agar. The optimum temperature and pH for the degradation of cypermethrin by immobilized cells of Micrococcus sp. were found to be 30 °C and 7.0, respectively. The rate of degradation of 10 and 20 mM of cypermethrin by freely suspended cells were compared with that of immobilized cells in batches and semi-continuous with shaken cultures. PUF-immobilized cells showed higher degradation of cypermethrin (10 mM and 20 mM) than freely suspended cells and cells immobilized in other matrices. The PUF-immobilized cells of Micrococcus sp. strain CPN 1 were retain their degradation capacity. Thus, they can be reused for more than 32 cycles, without losing their degradation capacity. Hence, the PUF-immobilized cells of Micrococcus sp. could potentially be used in the bioremediation of cypermethrin contaminated water.

  17. Non-encapsulated strains reveal novel insights in invasion and survival of Streptococcus suis in epithelial cells.

    Science.gov (United States)

    Benga, L; Goethe, R; Rohde, M; Valentin-Weigand, P

    2004-09-01

    Streptococcus suis is a porcine and human pathogen causing invasive diseases, such as meningitis or septicaemia. Host cell interactions of S. suis have been studied mainly with serotype 2 strains, but multiple capsular serotypes as well as non-typeable strains exist with diverse virulence features. At present, S. suis is considered an extracellular pathogen. However, whether or not it can also invade host cells is a matter of controversial discussions. We have assessed adherence and invasion of S. suis for HEp-2 epithelial cells by comparing 10 serotype 2 strains and four non-typeable (NT) strains. Only the NT strains and a non-encapsulated serotype 2 mutant strain, but none of the serotype 2 strains, adhered strongly and were invasive. Invasion seemed to be affected by environmental signals, as suggested from comparison of strains grown in different media. Further phenotypic and genotypic characterization revealed a high diversity among the different strains. Electron microscopic analysis of invasion of selected invasive NT strains indicated different uptake mechanisms. One strain induced large invaginations comparable to those seen in 'caveolae' mediated uptake, whereas invasion of the other strains was accompanied by formation of filipodia-like membrane protrusions. Invasion of all strains, however, was similarly susceptible to hypertonic sucrose, which inhibits receptor-mediated endocytosis. Irrespective of the uptake pathway, streptococci resided in acidified phago-lysosome like vacuoles. All strains, except one, survived intracellularly as well as extracellular acidic conditions. Survival seemed to be associated with the AdiS protein, an environmentally regulated arginine deiminase of S. suis. Concluding, invasion and survival of NT strains of S. suis in epithelial cells revealed novel evidence that S. suis exhibits a broad variety of virulence-associated features depending on genetic variation and regulation.

  18. Cell-surface alterations in class IIa bacteriocin-resistant Listeria monocytogenes strains.

    Science.gov (United States)

    Vadyvaloo, Viveka; Arous, Safia; Gravesen, Anne; Héchard, Yann; Chauhan-Haubrock, Ramola; Hastings, John W; Rautenbach, Marina

    2004-09-01

    Strains of the food-borne pathogen Listeria monocytogenes, showing either intermediate or high-level resistance to class IIa bacteriocins, were investigated to determine characteristics that correlated with their sensitivity levels. Two intermediate and one highly resistant spontaneous mutant of L. monocytogenes B73, a highly resistant mutant of L. monocytogenes 412, and a highly resistant, defined (mptA) mutant of L. monocytogenes EGDe were compared with their respective wild-type strains in order to investigate the contribution of different factors to resistance. Decreased mannose-specific phosphotransferase system gene expression (mptA, EIIAB(Man) component) was implicated in all levels of resistance, confirming previous studies by the authors' group. However, a clear correlation between d-alanine content in teichoic acid (TA), in particular the alanine : phosphorus ratio, and a more positive cell surface, as determined by cytochrome c binding, were found for the highly resistant strains. Furthermore, two of the three highly resistant strains showed a significant increase in sensitivity towards d-cycloserine (DCS). However, real-time PCR of the dltA (d-alanine esterification), and dal and ddlA genes (peptidoglycan biosynthesis) showed no change in transcriptional levels. The link between DCS sensitivity and increased d-alanine esterification of TA may be that DCS competes with alanine for transport via the alanine transporter. A possible tendency towards increased lysinylation of membrane phospholipid in the highly resistant strains was also found. A previous study reported that cell membranes of all the resistant strains, including the intermediate resistant strains, contained more unsaturated phosphatidylglycerol, which is an indication of a more fluid cell membrane. The results of that study correlate with the possible lysinylation, decreased mptA expression, d-alanine esterification of TA and more positive cell surface charge found in this study for

  19. Strain-Dependent Effect of Macroautophagy on Abnormally Folded Prion Protein Degradation in Infected Neuronal Cells.

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    Daisuke Ishibashi

    Full Text Available Prion diseases are neurodegenerative disorders caused by the accumulation of abnormal prion protein (PrPSc in the central nervous system. With the aim of elucidating the mechanism underlying the accumulation and degradation of PrPSc, we investigated the role of autophagy in its degradation, using cultured cells stably infected with distinct prion strains. The effects of pharmacological compounds that inhibit or stimulate the cellular signal transduction pathways that mediate autophagy during PrPSc degradation were evaluated. The accumulation of PrPSc in cells persistently infected with the prion strain Fukuoka-1 (FK, derived from a patient with Gerstmann-Sträussler-Scheinker syndrome, was significantly increased in cultures treated with the macroautophagy inhibitor 3-methyladenine (3MA but substantially reduced in those treated with the macroautophagy inducer rapamycin. The decrease in FK-derived PrPSc levels was mediated, at least in part, by the phosphatidylinositol 3-kinase/MEK signalling pathway. By contrast, neither rapamycin nor 3MA had any apparently effect on PrPSc from either the 22L or the Chandler strain, indicating that the degradation of PrPSc in host cells might be strain-dependent.

  20. Prediction of stress-strain behavior of ceramic matrix composites using unit cell model

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    Suzuki Takuya

    2015-01-01

    Full Text Available In this study, the elastic modulus and the stress-strain curve of ceramic matrix composites (CMCs were predicted by using the unit cell model that consists of fiber bundles and matrix. The unit cell model was developed based on the observation of cross sections of CMCs. The elastic modulus of CMCs was calculated from the results of finite element analysis using the developed model. The non-linear behavior of stress-strain curve of CMCs was also predicted by taking the degradation of the elastic modulus into consideration, where the degradation was related to the experimentally measured crack density in CMCs. The approach using the unit cell model was applied to two kinds of CMCs, and good agreement was obtained between the experimental and the calculated results.

  1. Influence of manufacturing processes on cell surface properties of probiotic strain Lactobacillus rhamnosus Lcr35®.

    Science.gov (United States)

    Nivoliez, Adrien; Veisseire, Philippe; Alaterre, Elina; Dausset, Caroline; Baptiste, Fabrice; Camarès, Olivier; Paquet-Gachinat, Marylise; Bonnet, Muriel; Forestier, Christiane; Bornes, Stéphanie

    2015-01-01

    The influence of the industrial process on the properties of probiotics, administered as complex manufactured products, has been poorly investigated. In the present study, we comparatively assessed the cell wall characteristics of the probiotic strain Lactobacillus rhamnosus Lcr35® together with three of its commercial formulations with intestinal applications. Putative secreted and transmembrane-protein-encoding genes were initially searched in silico in the genome of L. rhamnosus Lcr35®. A total of 369 candidate genes were identified which expressions were followed using a custom Lactobacillus DNA chip. Among them, 60 or 67 genes had their expression either upregulated or downregulated in the Lcr Restituo® packet or capsule formulations, compared to the native Lcr35® strain. Moreover, our data showed that the probiotic formulations (Lcr Lenio®, Lcr restituo® capsule and packet) showed a better capacity to adhere to intestinal epithelial Caco-2 cells than the native Lcr35® strain. Microbial (MATS) tests showed that the probiotic was an electron donor and that they were more hydrophilic than the native strain. The enhanced adhesion capacity of the active pharmaceutical ingredients (APIs) to epithelial Caco-2 cells and their antipathogen effect could be due to this greater surface hydrophilic character. These findings suggest that the manufacturing process influences the protein composition and the chemical properties of the cell wall. It is therefore likely that the antipathogen effect of the formulation is modulated by the industrial process. Screening of the manufactured products' properties would therefore represent an essential step in evaluating the effects of probiotic strains.

  2. Variable Inhibition of Zika Virus Replication by Different Wolbachia Strains in Mosquito Cell Cultures.

    Science.gov (United States)

    Schultz, Michaela J; Isern, Sharon; Michael, Scott F; Corley, Ronald B; Connor, John H; Frydman, Horacio M

    2017-07-15

    Mosquito-borne arboviruses are a major source of human disease. One strategy to reduce arbovirus disease is to reduce the mosquito's ability to transmit virus. Mosquito infection with the bacterial endosymbiont Wolbachia pipientis wMel is a novel strategy to reduce Aedes mosquito competency for flavivirus infection. However, experiments investigating cyclic environmental temperatures have shown a reduction in maternal transmission of wMel, potentially weakening the integration of this strain into a mosquito population relative to that of other Wolbachia strains. Consequently, it is important to investigate additional Wolbachia strains. All Zika virus (ZIKV) suppression studies are limited to the wMel Wolbachia strain. Here we show ZIKV inhibition by two different Wolbachia strains: wAlbB (isolated from Aedes albopictus mosquitoes) and wStri (isolated from the planthopper Laodelphax striatellus) in mosquito cells. Wolbachia strain wStri inhibited ZIKV most effectively. Single-cycle infection experiments showed that ZIKV RNA replication and nonstructural protein 5 translation were reduced below the limits of detection in wStri-containing cells, demonstrating early inhibition of virus replication. ZIKV replication was rescued when Wolbachia was inhibited with a bacteriostatic antibiotic. We observed a partial rescue of ZIKV growth when Wolbachia-infected cells were supplemented with cholesterol-lipid concentrate, suggesting competition for nutrients as one of the possible mechanisms of Wolbachia inhibition of ZIKV. Our data show that wAlbB and wStri infection causes inhibition of ZIKV, making them attractive candidates for further in vitro mechanistic and in vivo studies and future vector-centered approaches to limit ZIKV infection and spread.IMPORTANCE Zika virus (ZIKV) has swiftly spread throughout most of the Western Hemisphere. This is due in large part to its replication in and spread by a mosquito vector host. There is an urgent need for approaches that limit

  3. Matrix production and organization by endothelial colony forming cells in mechanically strained engineered tissue constructs.

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    Nicky de Jonge

    Full Text Available AIMS: Tissue engineering is an innovative method to restore cardiovascular tissue function by implanting either an in vitro cultured tissue or a degradable, mechanically functional scaffold that gradually transforms into a living neo-tissue by recruiting tissue forming cells at the site of implantation. Circulating endothelial colony forming cells (ECFCs are capable of differentiating into endothelial cells as well as a mesenchymal ECM-producing phenotype, undergoing Endothelial-to-Mesenchymal-transition (EndoMT. We investigated the potential of ECFCs to produce and organize ECM under the influence of static and cyclic mechanical strain, as well as stimulation with transforming growth factor β1 (TGFβ1. METHODS AND RESULTS: A fibrin-based 3D tissue model was used to simulate neo-tissue formation. Extracellular matrix organization was monitored using confocal laser-scanning microscopy. ECFCs produced collagen and also elastin, but did not form an organized matrix, except when cultured with TGFβ1 under static strain. Here, collagen was aligned more parallel to the strain direction, similar to Human Vena Saphena Cell-seeded controls. Priming ECFC with TGFβ1 before exposing them to strain led to more homogenous matrix production. CONCLUSIONS: Biochemical and mechanical cues can induce extracellular matrix formation by ECFCs in tissue models that mimic early tissue formation. Our findings suggest that priming with bioactives may be required to optimize neo-tissue development with ECFCs and has important consequences for the timing of stimuli applied to scaffold designs for both in vitro and in situ cardiovascular tissue engineering. The results obtained with ECFCs differ from those obtained with other cell sources, such as vena saphena-derived myofibroblasts, underlining the need for experimental models like ours to test novel cell sources for cardiovascular tissue engineering.

  4. Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida)

    Science.gov (United States)

    Rozas, Enrique E.; Albano, Rodolpho M.; Lôbo-Hajdu, Gisele; Müller, Werner E.G.; Schröder, Heinz-C.; Custódio, Márcio R.

    2011-01-01

    Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques. PMID:24031790

  5. Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida

    Directory of Open Access Journals (Sweden)

    Enrique E. Rozas

    2011-12-01

    Full Text Available Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs and single sponge cells (cytospins and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.

  6. Pathological cyclic strain-induced apoptosis in human periodontal ligament cells through the RhoGDIα/caspase-3/PARP pathway.

    Directory of Open Access Journals (Sweden)

    Li Wang

    Full Text Available AIM: Human periodontal ligament (PDL cells incur changes in morphology and express proteins in response to cyclic strain. However, it is not clear whether cyclic strain, especially excessive cyclic strain, induces PDL cell apoptosis and if so, what mechanism(s are responsible. The aim of the present study was to elucidate the molecular mechanisms by which pathological levels of cyclic strain induce human PDL cell apoptosis. MATERIALS AND METHODS: Human PDL cells were obtained from healthy premolar tissue. After three to five passages in culture, the cells were subjected to 20% cyclic strain at a frequency of 0.1 Hz for 6 or 24 h using an FX-5000T system. Morphological changes of the cells were assessed by inverted phase-contrast microscopy, and apoptosis was detected by fluorescein isothiocyanate (FITC-conjugated annexin V and propidium iodide staining followed by flow cytometry. Protein expression was evaluated by Western blot analysis. RESULTS: The number of apoptotic human PDL cells increased in a time-dependent manner in response to pathological cyclic strain. The stretched cells were oriented parallel to each another with their long axes perpendicular to the strain force vector. Cleaved caspase-3 and poly-ADP-ribose polymerase (PARP protein levels increased in response to pathological cyclic strain over time, while Rho GDP dissociation inhibitor alpha (RhoGDIα decreased. Furthermore, knock-down of RhoGDIα by targeted siRNA transfection increased stretch-induced apoptosis and upregulated cleaved caspase-3 and PARP protein levels. Inhibition of caspase-3 prevented stretch-induced apoptosis, but did not change RhoGDIα protein levels. CONCLUSION: The overall results suggest that pathological-level cyclic strain not only influenced morphology but also induced apoptosis in human PDL cells through the RhoGDIα/caspase-3/PARP pathway. Our findings provide novel insight into the mechanism of apoptosis induced by pathological cyclic strain in

  7. Proteomic Profiling of Mesenchymal Stem Cell Responses to Mechanical Strain and TGF-B1

    Energy Technology Data Exchange (ETDEWEB)

    Kurpinski, Kyle; Chu, Julia; Wang, Daojing; Li, Song

    2009-10-12

    Mesenchymal stem cells (MSCs) are a potential source of smooth muscle cells (SMCs) for constructing tissue-engineered vascular grafts. However, the details of how specific combinations of vascular microenvironmental factors regulate MSCs are not well understood. Previous studies have suggested that both mechanical stimulation with uniaxial cyclic strain and chemical stimulation with transforming growth factor {beta}1 (TGF-{beta}1) can induce smooth muscle markers in MSCs. In this study, we investigated the combined effects of uniaxial cyclic strain and TGF-{beta}1 stimulation on MSCs. By using a proteomic analysis, we found differential regulation of several proteins and genes, such as the up-regulation of TGF-{beta}1-induced protein ig-h3 (BGH3) protein levels by TGF-{beta}1 and up-regulation of calponin 3 protein level by cyclic strain. At the gene expression level, BGH3 was induced by TGF-{beta}1, but calponin 3 was not significantly regulated by mechanical strain or TGF-{beta}1, which was in contrast to the synergistic up-regulation of calponin 1 gene expression by cyclic strain and TGF-{beta}1. Further experiments with cycloheximide treatment suggested that the up-regulation of calponin 3 by cyclic strain was at post-transcriptional level. The results in this study suggest that both mechanical stimulation and TGF-{beta}1 signaling play unique and important roles in the regulation of MSCs at both transcriptional and post-transcriptional levels, and that a precise combination of microenvironmental cues may promote MSC differentiation.

  8. Dynamics of cell proliferation in the adult dentate gyrus of two inbred strains of mice

    Science.gov (United States)

    Hayes, N. L.; Nowakowski, R. S.

    2002-01-01

    The output potential of proliferating populations in either the developing or the adult nervous system is critically dependent on the length of the cell cycle (T(c)) and the size of the proliferating population. We developed a new approach for analyzing the cell cycle, the 'Saturate and Survive Method' (SSM), that also reveals the dynamic behaviors in the proliferative population and estimates of the size of the proliferating population. We used this method to analyze the proliferating population of the adult dentate gyrus in 60 day old mice of two inbred strains, C57BL/6J and BALB/cByJ. The results show that the number of cells labeled by exposure to BUdR changes dramatically with time as a function of the number of proliferating cells in the population, the length of the S-phase, cell division, the length of the cell cycle, dilution of the S-phase label, and cell death. The major difference between C57BL/6J and BALB/cByJ mice is the size of the proliferating population, which differs by a factor of two; the lengths of the cell cycle and the S-phase and the probability that a newly produced cell will die within the first 10 days do not differ in these two strains. This indicates that genetic regulation of the size of the proliferating population is independent of the genetic regulation of cell death among those newly produced cells. The dynamic changes in the number of labeled cells as revealed by the SSM protocol also indicate that neither single nor repeated daily injections of BUdR accurately measure 'proliferation.'.

  9. Protein Expression of BLM Gene and Its Apoptosis Sensitivity in Hematopoietic Tumor Cell Strains

    Institute of Scientific and Technical Information of China (English)

    Xiaobei WANG; Lihua HU

    2008-01-01

    Patients with Bloom syndrome (BS) show an immunodeficiency, an enhanced sister chromatid exchanges (SCEs), a strong genetic instability and an increased predisposition to all. In order to investigate the differential expression of BLM protein in hematopoietic tumor cell strains and study the effects of BLM gene on ultraviolet (UV)- or hydroxyurea (HU)-induced apoptosis, Western blot was used to detect the expression of BLM protein in normal human bone marrow mononuclear cells and 4 kinds of hematopoietic tumor cell strains. The 4 kinds of hematopoietic tumor cells were exposed to UV light with a germicidal UV lamp or treated with 2 mmol/L hydroxyurea and the apoptotic rate was detected by using AnnexinV-FITC. The results showed that these tumor cells ex- pressed BLM protein higher than the normal human bone marrow mononuclear cells (P<0.01). In the 4 hematopoietic tumor cells, BLM protein was all specially cleaved in response to UV- or HU-induced apoptosis. The increase of BLM protein expression may play an important role in the evelopment of these tumors, and BLM proteolysis is likely to be a general feature of the apoptotic esponse.

  10. Acinetobacter baumannii biofilms: variations among strains and correlations with other cell properties.

    Science.gov (United States)

    McQueary, Christin N; Actis, Luis A

    2011-04-01

    Acinetobacter baumannii is an opportunistic pathogen that causes serious infections in humans by colonizing and persisting on surfaces normally found in hospital settings. The capacity of this pathogen to persist in these settings could be due to its ability to form biofilms on inanimate surfaces. This report shows that although the ATCC 19606(T) type strain and 8 different clinical isolates form biofilms, there are significant variations in the cell density and microscopic structures of these cell aggregates, with 3 of the isolates forming pellicles floating on the surface of stagnant broth cultures. PCR indicated that, like ATCC 19606(T), all 8 clinical isolates harbor all the genetic components of the CsuA/BABCDE chaperone-usher pili assembly system, which is needed for biofilm formation on plastic. Pili detection in cells of all strains examined supports the presence and function of a pilus assembly system. However, only one of them produced the putative ATCC 19606(T) CsuA/B pilin subunit protein. Hydrophobicity tests and motility assays also showed significant variations among all tested strains and did not result in direct correlations between the biofilm phenotype and cell properties that could affect biofilm formation on abiotic surfaces. This lack of correlation among these 3 phenotypes may reflect some of the variations already reported with this pathogen, which may pose a challenge in the treatment of the infections this pathogen causes in humans using biofilm formation on abiotic surfaces as a target.

  11. Engineering Pichia pastoris for improved NADH regeneration: A novel chassis strain for whole-cell catalysis

    Directory of Open Access Journals (Sweden)

    Martina Geier

    2015-09-01

    Full Text Available Many synthetically useful reactions are catalyzed by cofactor-dependent enzymes. As cofactors represent a major cost factor, methods for efficient cofactor regeneration are required especially for large-scale synthetic applications. In order to generate a novel and efficient host chassis for bioreductions, we engineered the methanol utilization pathway of Pichia pastoris for improved NADH regeneration. By deleting the genes coding for dihydroxyacetone synthase isoform 1 and 2 (DAS1 and DAS2, NADH regeneration via methanol oxidation (dissimilation was increased significantly. The resulting Δdas1 Δdas2 strain performed better in butanediol dehydrogenase (BDH1 based whole-cell conversions. While the BDH1 catalyzed acetoin reduction stopped after 2 h reaching ~50% substrate conversion when performed in the wild type strain, full conversion after 6 h was obtained by employing the knock-out strain. These results suggest that the P. pastoris Δdas1 Δdas2 strain is capable of supplying the actual biocatalyst with the cofactor over a longer reaction period without the over-expression of an additional cofactor regeneration system. Thus, focusing the intrinsic carbon flux of this methylotrophic yeast on methanol oxidation to CO2 represents an efficient and easy-to-use strategy for NADH-dependent whole-cell conversions. At the same time methanol serves as co-solvent, inductor for catalyst and cofactor regeneration pathway expression and source of energy.

  12. Brain immune cell composition and functional outcome after cerebral ischemia: Comparison of two mouse strains

    Directory of Open Access Journals (Sweden)

    Hyun Ah eKim

    2014-11-01

    Full Text Available Inflammatory cells may contribute to secondary brain injury following cerebral ischemia. The C57Bl/6 mouse strain is known to exhibit a T helper 1-prone, pro-inflammatory type response to injury, whereas the FVB strain is relatively T helper 2-prone, or anti-inflammatory, in its immune response. We tested whether stroke outcome is more severe in C57Bl/6 than FVB mice. Male mice of each strain underwent sham surgery or 1 h occlusion of the middle cerebral artery followed by 23 h of reperfusion. Despite no difference in infarct size, C57Bl/6 mice displayed markedly greater functional deficits than FVB mice after stroke, as assessed by neurological scoring and hanging wire test. Total numbers of CD45+ leukocytes tended to be larger in the brains of C57Bl/6 than FVB mice after stroke, but there were marked differences in leukocyte composition between the two mouse strains. The inflammatory response in C57Bl/6 mice primarily involved T and B lymphocytes, whereas neutrophils, monocytes and macrophages were more prominent in FVB mice. Our data are consistent with the concept that functional outcome after stroke is dependent on the immune cell composition which develops following ischemic brain injury.

  13. Cyclic mechanical strain maintains Nanog expression through PI3K/Akt signaling in mouse embryonic stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Horiuchi, Rie [Division of Regenerative Medical Engineering, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033 (Japan); Akimoto, Takayuki, E-mail: akimoto@m.u-tokyo.ac.jp [Division of Regenerative Medical Engineering, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033 (Japan); Institute for Biomedical Engineering, Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 513 Waseda-tsurumaki, Shinjuku, Tokyo 162-0041 (Japan); Hong, Zhang [Institute for Biomedical Engineering, Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 513 Waseda-tsurumaki, Shinjuku, Tokyo 162-0041 (Japan); Ushida, Takashi [Division of Regenerative Medical Engineering, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-0033 (Japan)

    2012-08-15

    Mechanical strain has been reported to affect the proliferation/differentiation of many cell types; however, the effects of mechanotransduction on self-renewal as well as pluripotency of embryonic stem (ES) cells remains unknown. To investigate the effects of mechanical strain on mouse ES cell fate, we examined the expression of Nanog, which is an essential regulator of self-renewal and pluripotency as well as Nanog-associated intracellular signaling during uniaxial cyclic mechanical strain. The mouse ES cell line, CCE was plated onto elastic membranes, and we applied 10% strain at 0.17 Hz. The expression of Nanog was reduced during ES cell differentiation in response to the withdrawal of leukemia inhibitory factor (LIF); however, two days of cyclic mechanical strain attenuated this reduction of Nanog expression. On the other hand, the cyclic mechanical strain promoted PI3K-Akt signaling, which is reported as an upstream of Nanog transcription. The cyclic mechanical strain-induced Akt phosphorylation was blunted by the PI3K inhibitor wortmannin. Furthermore, cytochalasin D, an inhibitor of actin polymerization, also inhibited the mechanical strain-induced increase in phospho-Akt. These findings imply that mechanical force plays a role in regulating Nanog expression in ES cells through the actin cytoskeleton-PI3K-Akt signaling. -- Highlights: Black-Right-Pointing-Pointer The expression of Nanog, which is an essential regulator of 'stemness' was reduced during embryonic stem (ES) cell differentiation. Black-Right-Pointing-Pointer Cyclic mechanical strain attenuated the reduction of Nanog expression. Black-Right-Pointing-Pointer Cyclic mechanical strain promoted PI3K-Akt signaling and mechanical strain-induced Akt phosphorylation was blunted by the PI3K inhibitor and an inhibitor of actin polymerization.

  14. Poor Invasion of Trophoblastic Cells but Normal Plaque Formation in Fibroblastic Cells despite actA Deletion in a Group of Listeria monocytogenes Strains Persisting in Some Food Processing Environments

    DEFF Research Database (Denmark)

    Holch, Anne; Gottlieb, Caroline Trebbien; Larsen, Marianne Halberg

    2010-01-01

    L. monocytogenes strains, including clinical strains, and they carry a premature stop codon in inlA. Eight of 15 strains, including the RAPD 9 and maternofetal strains, had a 105-nucleotide deletion in actA that did not affect cell-to-cell spread in mouse fibroblasts. The RAPD 9 strains may still...

  15. Characteristics of Cell-mediated, Anti-listerial Immunity Induced by A Naturally Avirulent Listeria monocytogenes Serotype 4a Strain HCC23

    Science.gov (United States)

    The characteristics of cell-mediated, anti-listerial immune response initiated by an avirulent Listeria monocytogenes serotype 4a strain HCC23 was assessed. Similar to virulent strain EGD, avirulent strain HCC23 grew readily within macrophage-like J774 cells, but nonhemolytic strain ATCC 15313 did n...

  16. Doxorubicin Cardiotoxicity and Cardiac Function Improvement After Stem Cell Therapy Diagnosed by Strain Echocardiography

    OpenAIRE

    Oliveira, Maira S.; Melo, Marcos B; Carvalho, Juliana L; Melo, Isabela M; Lavor, Mario SL; Gomes, Dawidson A.; Goes, Alfredo M.; Melo, Marilia M

    2013-01-01

    Doxorubicin (Dox) is one of the most effective chemotherapeutic agents; however, it causes dose-dependent cardiotoxicity. Evaluation of left ventricular function relies on measurements based on M-mode echocardiography. A new technique based on quantification of myocardial motion and deformation, strain echocardiography, has been showed promising profile for early detection of cardiac dysfunction. Different therapy strategies, such as flavonoid plant extracts and stem cells, have been investig...

  17. Cell wall degrading isoenzyme profiles of Trichoderma biocontrol strains show correlation with rDNA species

    Institute of Scientific and Technical Information of China (English)

    Sanz L; Hermosa M R; González F J; Monte E

    2004-01-01

    @@ Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE).Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderna.Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique.Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities.Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.

  18. Scanning electron microscopic study of human neuroblastoma cells affected with Naegleria fowleri Thai strains.

    Science.gov (United States)

    Tiewcharoen, Supathra; Rabablert, Jundee; Chetanachan, Pruksawan; Junnu, Virach; Worawirounwong, Dusit; Malainual, Nat

    2008-10-01

    In order to understand the pathogenesis of Naegleria fowleri in primary amoebic meningoencephalitis, the human neuroblastoma (SK-N-MC) and African green monkey kidney (Vero) cells were studied in vitro. Amoeba suspension in cell-culture medium was added to the confluent monolayer of SK-N-MC and Vero cells. The cytopathic activity of N. fowleri trophozoites in co-culture system was elucidated by scanning electron microscope at 3, 6, 9, 12, and 24 h. Two strains of N. fowleri displayed well-organized vigorous pseudopods in Nelson's medium at 37 degrees C. In co-culture, the target monolayer cells were damaged by two mechanisms, phagocytosis by vigorous pseudopods and engulfment by sucker-like apparatus. N. fowleri trophozoites produced amoebostomes only in co-culture with SK-N-MC cells. In contrast, we could not find such apparatus in the co-culture with Vero cells. The complete destruction time (100%) at 1:1 amoeba/cells ratio of SK-N-MC cells (1 day) was shorter than the Vero cells (12 days). In conclusion, SK-N-MC cells were confirmed to be a target model for studying neuropathogenesis of primary amoebic meningoencephalitis.

  19. Immobilization of cells with nitrilase activity from a thermophilic bacterial strain.

    Science.gov (United States)

    Kabaivanova, L; Dobreva, E; Dimitrov, P; Emanuilova, E

    2005-01-01

    Cells of the moderately thermophilic Bacillus sp. UG-5B strain, producing nitrilase (EC3.5.5.1), which converts nitriles directly to the corresponding acid and ammonia, were immobilized using different types of matrices and techniques. A variety of sol-gel silica hybrids were tested for entrapment and adsorption of bacterial cells as well as chemical binding on polysulphone membranes. Activation of the matrix surface with formaldehyde led to an increase in immobilization efficiency and operational stability of the biocatalysts. Among the supports screened, membranes gave the best results for enzyme activity and especially operational stability, with retention of 100% activity after eight reaction cycles.

  20. Thermal isocreep curves obtained during multi-axial creep tests on recrystallized Zircaloy-4 and M5™ alloy

    Energy Technology Data Exchange (ETDEWEB)

    Rautenberg, M., E-mail: mrautenb@gmail.com [AREVA, AREVA NP, 10 rue Juliette Récamier, 69456 Lyon (France); CIRIMAT, CNRS/UPS/INPT, 4 allée Emile Monso, 31030 Toulouse (France); Poquillon, D. [CIRIMAT, CNRS/UPS/INPT, 4 allée Emile Monso, 31030 Toulouse (France); Pilvin, P. [LIMATB, University Bretagne-Sud, rue de Saint-Maudé, 56321 Lorient (France); Grosjean, C. [AREVA, AREVA NP, 10 rue Juliette Récamier, 69456 Lyon (France); CIRIMAT, CNRS/UPS/INPT, 4 allée Emile Monso, 31030 Toulouse (France); Cloué, J.M. [AREVA, AREVA NP, 10 rue Juliette Récamier, 69456 Lyon (France); Feaugas, X. [LEMMA, Université de La Rochelle, Avenue Michel Crépeau, 17042 La Rochelle (France)

    2014-04-01

    Zirconium alloys are widely used in the nuclear industry. Several components, such as cladding or guide tubes, undergo strong mechanical loading during and after their use inside the pressurized water reactors. The current requirements on higher fuel performances lead to the developing on new Zr based alloys exhibiting better mechanical properties. In this framework, creep behaviors of recrystallized Zircaloy-4 and M5™, have been investigated and then compared. In order to give a better understanding of the thermal creep anisotropy of Zr-based alloys, multi-axial creep tests have been carried out at 673 K. Using a specific device, creep conditions have been set using different values of β = σ{sub zz}/σ{sub θθ}, σ{sub zz} and σ{sub θθ} being respectively the axial and hoop creep stresses. Both axial and hoop strains are measured during each test which is carried out until stationary creep is stabilized. The steady-state strain rates are then used to build isocreep curves. Considering the isocreep curves, the M5™ alloy shows a largely improved creep resistance compared to the recrystallized Zircaloy-4, especially for tubes under high hoop loadings (0 < β < 1). The isocreep curves are then compared with simulations performed using two different mechanical models. Model 1 uses a von Mises yield criterion, the model 2 is based on a Hill yield criterion. For both models, a coefficient derived from Norton law is used to assess the stress dependence.

  1. Role of calcium signaling in down-regulation of aggrecan induced by cyclic tensile strain in annulus fibrosus cells

    Institute of Scientific and Technical Information of China (English)

    GUO Zhi-liang; ZHOU Yue; LI Hua-zhuang; CAO Guo-yong; TENG Hai-jun

    2006-01-01

    Objective:To study the role of intracellular calcium signal pathway in the down-regulation of aggrecan induced by cyclic tensile strain in the annulus fibrosus cells. Methods :The expression of aggrecan mRNA and core protein were respectively detected with RT-PCR and western blot after the channels transmitting calcium ions were blocked with EGTA, gadolinium and verapamil. Results:EGTA, gadolinium and verapamil partially prevented the effects of cyclic tensile strain on the expression of aggrecan in annulus fibrosus cells. Conclusion:The calcium signaling is involved in the down-regulation of proteoglycan resulting from cyclic tensile strain in the annulus fibrosus cells.

  2. Mouse Reporter Strain for Noninvasive Bioluminescent Imaging of Cells that have Undergone Cre-Mediated Recombination

    Directory of Open Access Journals (Sweden)

    Michal Safran

    2003-10-01

    Full Text Available Conditional alleles containing LoxP recombination sites, in conjunction with Cre recombinase delivered by a variety of means, allows for spatial and temporal control of gene expression in mouse models. Here we describe a mouse strain in which a luciferase (Luc cDNA, preceded by a LoxP-stop-LoxP (L-S-L cassette, was introduced into the ubiquitously expressed ROSA26 locus. Mouse embryo fibroblasts derived from this strain expressed luciferase after Cre-mediated recombination in vitro. ROSA26 L-S-L-Luc/+ mice expressed luciferase in a diffuse or liver-restricted pattern, as determined by noninvasive, bioluminescent imaging, when crossed to transgenic mice in which Cre was under the control of a zygotically expressed (EIIA-Cre, or a liver-restricted (albumin-Cre, promoter, respectively. Organ-specific luciferase expression was also seen after intraparenchymal administration of an adenovirus encoding Cre. The ROSA26 L-S-L-Luc/+ strain should be useful for characterizing Cre mouse strains and for following the fate of cells that have undergone Cre-mediated recombination in vivo.

  3. Multiplicative earthquake likelihood models incorporating strain rates

    Science.gov (United States)

    Rhoades, D. A.; Christophersen, A.; Gerstenberger, M. C.

    2017-01-01

    SUMMARYWe examine the potential for strain-rate variables to improve long-term earthquake likelihood models. We derive a set of multiplicative hybrid earthquake likelihood models in which cell rates in a spatially uniform baseline model are scaled using combinations of covariates derived from earthquake catalogue data, fault data, and strain-rates for the New Zealand region. Three components of the strain rate estimated from GPS data over the period 1991-2011 are considered: the shear, rotational and dilatational strain rates. The hybrid model parameters are optimised for earthquakes of M 5 and greater over the period 1987-2006 and tested on earthquakes from the period 2012-2015, which is independent of the strain rate estimates. The shear strain rate is overall the most informative individual covariate, as indicated by Molchan error diagrams as well as multiplicative modelling. Most models including strain rates are significantly more informative than the best models excluding strain rates in both the fitting and testing period. A hybrid that combines the shear and dilatational strain rates with a smoothed seismicity covariate is the most informative model in the fitting period, and a simpler model without the dilatational strain rate is the most informative in the testing period. These results have implications for probabilistic seismic hazard analysis and can be used to improve the background model component of medium-term and short-term earthquake forecasting models.

  4. Flocculation in ale brewing strains of Saccharomyces cerevisiae: re-evaluation of the role of cell surface charge and hydrophobicity.

    Science.gov (United States)

    Holle, Ann Van; Machado, Manuela D; Soares, Eduardo V

    2012-02-01

    Flocculation is an eco-friendly process of cell separation, which has been traditionally exploited by the brewing industry. Cell surface charge (CSC), cell surface hydrophobicity (CSH) and the presence of active flocculins, during the growth of two (NCYC 1195 and NCYC 1214) ale brewing flocculent strains, belonging to the NewFlo phenotype, were examined. Ale strains, in exponential phase of growth, were not flocculent and did not present active flocculent lectins on the cell surface; in contrast, the same strains, in stationary phase of growth, were highly flocculent (>98%) and presented a hydrophobicity of approximately three to seven times higher than in exponential phase. No relationship between growth phase, flocculation and CSC was observed. For comparative purposes, a constitutively flocculent strain (S646-1B) and its isogenic non-flocculent strain (S646-8D) were also used. The treatment of ale brewing and S646-1B strains with pronase E originated a loss of flocculation and a strong reduction of CSH; S646-1B pronase E-treated cells displayed a similar CSH as the non-treated S646-8D cells. The treatment of the S646-8D strain with protease did not reduce CSH. In conclusion, the increase of CSH observed at the onset of flocculation of ale strains is a consequence of the presence of flocculins on the yeast cell surface and not the cause of yeast flocculation. CSH and CSC play a minor role in the auto-aggregation of the ale strains since the degree of flocculation is defined, primarily, by the presence of active flocculins on the yeast cell wall.

  5. Cyclic mechanical strain-induced proliferation and migration of human airway smooth muscle cells: role of EMMPRIN and MMPs.

    Science.gov (United States)

    Hasaneen, Nadia A; Zucker, Stanley; Cao, Jian; Chiarelli, Christian; Panettieri, Reynold A; Foda, Hussein D

    2005-09-01

    Airway smooth muscle (ASM) proliferation and migration are major components of airway remodeling in asthma. Asthmatic airways are exposed to mechanical strain, which contributes to their remodeling. Matrix metalloproteinase (MMP) plays an important role in remodeling. In the present study, we examined if the mechanical strain of human ASM (HASM) cells contributes to their proliferation and migration and the role of MMPs in this process. HASM were exposed to mechanical strain using the FlexCell system. HASM cell proliferation, migration and MMP release, activation, and expression were assessed. Our results show that cyclic strain increased the proliferation and migration of HASM; cyclic strain increased release and activation of MMP-1, -2, and -3 and membrane type 1-MMP; MMP release was preceded by an increase in extracellular MMP inducer; Prinomastat [a MMP inhibitor (MMPI)] significantly decreased cyclic strain-induced proliferation and migration of HASM; and the strain-induced increase in the release of MMPs was accompanied by an increase in tenascin-C release. In conclusion, cyclic mechanical strain plays an important role in HASM cell proliferation and migration. This increase in proliferation and migration is through an increase in MMP release and activation. Pharmacological MMPIs should be considered in the pursuit of therapeutic options for airway remodeling in asthma.

  6. Selected Lactobacillus strains isolated from sugary and milk kefir reduce Salmonella infection of epithelial cells in vitro.

    Science.gov (United States)

    Zavala, L; Golowczyc, M A; van Hoorde, K; Medrano, M; Huys, G; Vandamme, P; Abraham, A G

    2016-09-01

    The isolation of potentially probiotic strains and the subsequent study of their properties are very important steps to gain insight in the health benefits ascribed to sugary and milk kefir. The aim of the present study was to characterise fifteen Lactobacillus strains isolated from these beverages by determining some surface properties and their ability to antagonise enterocyte cell damage after Salmonella infection in vitro. Lactobacillus surface properties were determined by hydrophobicity, autoaggregation, and coaggregation assays with Salmonella. In addition, lactobacilli adhesion to Caco-2/TC-7 cells and the effect on Salmonella invasion were evaluated. Finally, the disassembly of F-actin cytoskeleton on intestinal epithelial cells was assayed in vitro when Salmonella infection was performed in the presence of selected Lactobacillus strains. Ten out of the 15 strains showed a high adhesion capacity to Caco-2/TC-7 cells. Most of the strains were hydrophilic and non-autoaggregating. Strains isolated from sugary kefir were non-coaggregating with Salmonella, while strains Lactobacillus paracasei CIDCA 83120, 83121, 83123, 83124, 8339, 83102 isolated from milk kefir were able to coaggregate after 1 h. L. paracasei CIDCA 8339 and Lactobacillus kefiri CIDCA 83102 were able to diminish Salmonella invasion to the enterocytes. An antagonistic effect on cytoskeleton disruption elicited by the pathogen was also demonstrated. Our results suggest that both strains isolated from milk kefir could be considered as appropriate probiotic candidates.

  7. Activation of the adenylyl cyclase/cyclic AMP/protein kinase A pathway in endothelial cells exposed to cyclic strain

    Science.gov (United States)

    Cohen, C. R.; Mills, I.; Du, W.; Kamal, K.; Sumpio, B. E.

    1997-01-01

    The aim of this study was to assess the involvement of the adenylyl cyclase/cyclic AMP/protein kinase A pathway (AC) in endothelial cells (EC) exposed to different levels of mechanical strain. Bovine aortic EC were seeded to confluence on flexible membrane-bottom wells. The membranes were deformed with either 150 mm Hg (average 10% strain) or 37.5 mm Hg (average 6% strain) vacuum at 60 cycles per minute (0.5 s strain; 0.5 s relaxation) for 0-60 min. The results demonstrate that at 10% average strain (but not 6% average strain) there was a 1.5- to 2.2-fold increase in AC, cAMP, and PKA activity by 15 min when compared to unstretched controls. Further studies revealed an increase in cAMP response element binding protein in EC subjected to the 10% average strain (but not 6% average strain). These data support the hypothesis that cyclic strain activates the AC/cAMP/PKA signal transduction pathway in EC which may occur by exceeding a strain threshold and suggest that cyclic strain may stimulate the expression of genes containing cAMP-responsive promoter elements.

  8. Analysis and simulation of high strain compression of anisotropic open-cell elastic foams

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    By elongating the regular Kelvin model in one direction and keeping unchanged in the other two directions,the anisotropic model was constructed.Then,the simplified periodic structural cell was obtained according to the periodicity and symmetry of the model in the whole space.Using the half-strut element and elastic deflection theory to analyze the mechanical behavior as were adopted in the previous studies,this paper obtained the theoretical expressions for the compressive stress and strain as well as the corresponding curves in the rise and transverse directions.In addition,the theoretical results were examined by the finite element simulation.Results indicated that the theoretical analysis was very close to the finite element simulation when the strain was not too high,which confirmed the validity of theoretical analysis.At the same time,the anisotropy was shown to have a significant effect on the mechanical properties of open-cell foams.As the anisotropy ratio increased,the compressive stress was improved in the rise direction but dropped in the transverse direction under the same strain.

  9. Human endothelial cells allow passage of an archetypal BK virus (BKV) strain--a tool for cultivation and functional studies of natural BKV strains.

    Science.gov (United States)

    Hanssen Rinaldo, C; Hansen, H; Traavik, T

    2005-07-01

    The ubiquitous human polyomavirus BK (BKV) causes the serious condition BKV-nephropathy in an increasing number of renal-transplant patients. The lack of authentic cell cultures for multiplication of naturally occurring strains has hampered cultivation and functional studies of BKV. Here we demonstrate that the most common urine shed BKV strain, the archetype, multiplies in the human endothelial cell line HUV-EC-C. Additional variants with deletions in the non-coding control region (NCCR) appear upon prolonged propagation. Although the titer produced was low, at the present HUV-EC-C is the only cell line shown to allow propagation of archetypal BKV. HUV-EC-C may therefore be a useful tool for BKV cultivation as well as functional studies.

  10. High immunogenic enterovirus 71 strain and its production using serum-free microcarrier Vero cell culture.

    Science.gov (United States)

    Liu, Chia-Chyi; Lian, Wei-Cheng; Butler, Michael; Wu, Suh-Chin

    2007-01-01

    Developing an effective vaccine against enterovirus 71 (EV71) infection provides the best means to control the disease. We have previously reported that large-scale preparation of a low immunogenic EV71 strain can be achieved using serum free microcarrier Vero cell culture in a 2-l bioreactor [Wu SC, Liu CC, Lian WC. Optimization of microcarrier cell culture process for the inactivated enterovirus type 71 vaccine development. Vaccine 2004;22:3858-64]. This present work further investigated the virus growth and the immunogenicity of two high immunogenic strains (EV71-075 and EV71-117) prepared in serum-free microcarrier cell cultures. Our results showed that serum free culture increased cell death rate after infection, reduced the virus specific productivity, but resulted in elicitation of higher neutralizing titers in immunized mice as compared to that parallel obtained in serum-containing cultures. Therefore, serum-free microcarrier culture is a valuable process for developing inactivated EV71 vaccines.

  11. Probiotic potential and biotherapeutic effects of newly isolated vaginal Lactobacillus acidophilus 36YL strain on cancer cells.

    Science.gov (United States)

    Nami, Yousef; Abdullah, Norhafizah; Haghshenas, Babak; Radiah, Dayang; Rosli, Rozita; Khosroushahi, Ahmad Yari

    2014-08-01

    Lactobacillus acidophilus is categorized as a probiotic strain because of its beneficial effects in human health and prevention of disease transmission. This study is aimed to characterize the probiotic potential of L. acidophilus 36YL originally isolated from the vagina of healthy and fertile Iranian women. The L. acidophilus 36YL strain was identified using 16S rDNA gene sequencing and characterized by biochemical methodologies, such as antibiotics susceptibility, antimicrobial activity, and acid and bile resistance. The bioactivity of the secretion of this strain on four human cancer cell lines (AGS, HeLa, MCF-7, and HT-29) and one normal cell line (HUVEC) was evaluated by cytotoxicity assay and apoptosis analysis. This newly isolated strain was found to exhibit notable probiotic properties, such as admirable antibiotic susceptibility, good antimicrobial activity, and favorable resistance to acid and bile salt. The results of bioactivity assessment demonstrated acceptable anticancer effects on the four tested cancer cell lines and negligible side effects on the assayed normal cell line. Our findings revealed that the anticancer effect of L. acidophilus 36YL strain secretions depends on the induction of apoptosis in cancer cells. L. acidophilus 36YL strain is considered as a nutraceutical alternative or a topical medication with a potential therapeutic index because of the absence of cytotoxicity to normal cells, but effective toxicity to cancer cell lines.

  12. In Silico Constraint-Based Strain Optimization Methods: the Quest for Optimal Cell Factories.

    Science.gov (United States)

    Maia, Paulo; Rocha, Miguel; Rocha, Isabel

    2016-03-01

    Shifting from chemical to biotechnological processes is one of the cornerstones of 21st century industry. The production of a great range of chemicals via biotechnological means is a key challenge on the way toward a bio-based economy. However, this shift is occurring at a pace slower than initially expected. The development of efficient cell factories that allow for competitive production yields is of paramount importance for this leap to happen. Constraint-based models of metabolism, together with in silico strain design algorithms, promise to reveal insights into the best genetic design strategies, a step further toward achieving that goal. In this work, a thorough analysis of the main in silico constraint-based strain design strategies and algorithms is presented, their application in real-world case studies is analyzed, and a path for the future is discussed.

  13. Hantavirus infection suppresses thrombospondin-1 expression in cultured endothelial cells in a strain-specific manner

    Directory of Open Access Journals (Sweden)

    Svetlana F Khaiboullina

    2016-07-01

    Full Text Available Hantavirus infection is associated with two frequently fatal diseases in humans: hemorrhagic fever with renal syndrome (HFRS and hantavirus pulmonary syndrome (HPS. The pathogenesis of hantavirus infection is complex and not fully understood; however, it is believed to involve virus-induced hyperinflammatory immune responses. Thrombospondin-1 (THBS1 is a large homotrimeric protein that plays a putative role in regulating blood homeostasis. Hyperresponsiveness to inflammatory stimuli has also been associated with defects in the THBS1 gene. Our data suggest that hantavirus infection of human umbilical cord vein endothelial cells (HUVEC suppress the accumulation of THBS1 in the extracellular matrix. Additionally, this suppression is dependent on virus replication, implying a direct mechanism of action. Our data also imply that the pathogenic Andes and Hantaan strains inhibit THBS1 expression while the non-pathogenic Prospect Hill strain showed little inhibition. These observations suggest that a dysregulation of THBS1 may contribute to the pathogenesis of hantavirus infection.

  14. An ultra fast detection method reveals strain-induced Ca(2+) entry via TRPV2 in alveolar type II cells.

    Science.gov (United States)

    Fois, Giorgio; Wittekindt, Oliver; Zheng, Xing; Felder, Erika Tatiana; Miklavc, Pika; Frick, Manfred; Dietl, Paul; Felder, Edward

    2012-09-01

    A commonly used technique to investigate strain-induced responses of adherent cells is culturing them on an elastic membrane and globally stretching the membrane. However, it is virtually impossible to acquire microscopic images immediately after the stretch with this method. Using a newly developed technique, we recorded the strain-induced increase of the cytoplasmic Ca(2+) concentration ([Ca(2+)](c)) in rat primary alveolar type II (ATII) cells at an acquisition rate of 30ms and without any temporal delay. We can show that the onset of the mechanically induced rise in [Ca(2+)](c) was very fast (<30 ms), and Ca(2+) entry was immediately abrogated when the stimulus was withdrawn. This points at a direct mechanical activation of an ion channel. RT-PCR revealed high expression of TRPV2 in ATII cells, and silencing TRPV2, as well as blocking TRPV channels with ruthenium red, significantly reduced the strain-induced Ca(2+) response. Moreover, the usually homogenous pattern of the strain-induced [Ca(2+)](c) increase was converted into a point-like response after both treatments. Also interfering with actin/myosin and integrin binding inhibited the strain-induced increase of [Ca(2)](c). We conclude that TRPV2 participates in strain-induced Ca(2+) entry in ATII cells and suggest a direct mechanical activation of the channel that depends on FAs and actin/myosin. Furthermore, our results underline the importance of cell strain systems that allow high temporal resolution.

  15. Evidence for strain-specific differences in benzene toxicity as a function of host target cell susceptibility.

    Science.gov (United States)

    Neun, D J; Penn, A; Snyder, C A

    1992-01-01

    It has long been recognized that benzene exposure produces disparate toxic responses among different species or even among different strains within the same species. There is ample evidence that species- or strain-dependent differences in metabolic activity correlate with the disparate responses to benzene. However, bone marrow cells (the putative targets of benzene toxicity) may also exhibit species- or strain-dependent differences in susceptibility to the toxic effects of benzene. To investigate this hypothesis, two sets of companion experiments were performed. First, two strains of mice, Swiss Webster (SW) and C57B1/6J (C57), were exposed to 300 ppm benzene via inhalation and the effects of the exposures were determined on bone marrow cellularity and the development of bone marrow CFU-e (Colony Forming Unit-erythroid, an early red cell progenitor). Second, bone marrow cells from the same strains were exposed in vitro to five known benzene metabolites (1,4 benzoquinone, catechol, hydroquinone, muconic acid, and phenol) individually and in binary combinations. Benzene exposure, in vivo, reduced bone marrow cellularity and the development of CFU-e in both strains; however, reductions in both these endpoints were more severe in the SW strain. When bone marrow cells from the two strains were exposed in vitro to the five benzene metabolites individually, benzoquinone, hydroquinone, and catechol reduced the numbers of CFU-e in both strains in dose-dependent responses, phenol weakly reduced the numbers of the C57 CFU-e only and in a non-dose-dependent manner, and muconic acid was without effect on cells from either strain.(ABSTRACT TRUNCATED AT 250 WORDS)

  16. Inhibition of Aspergillus parasiticus and cancer cells by marine actinomycete strains

    Science.gov (United States)

    Li, Ping; Yan, Peisheng

    2014-12-01

    Ten actinomycete strains isolated from the Yellow Sea off China's coasts were identified as belonging to two genera by 16S rDNA phylogenetic analysis: Streptomyces and Nocardiopsis. Six Streptomyces strains (MA10, 2SHXF01-3, MA35, MA05-2, MA05-2-1 and MA08-1) and one Nocardiopsis strain (MA03) were predicted to have the potential to produce aromatic polyketides based on the analysis of the KSα (ketoacyl-synthase) gene in the type II PKS (polyketides synthase) gene cluster. Four strains (MA03, MA01, MA10 and MA05-2) exhibited significant inhibitory effects on mycelia growth (inhibition rate >50%) and subsequent aflatoxin production (inhibition rate >75%) of the mutant aflatoxigenic Aspergillus parasiticus NFRI-95. The ethyl acetate extracts of the broth of these four strains displayed significant inhibitory effects on mycelia growth, and the IC50 values were calculated (MA03: 0.275 mg mL-1, MA01: 0.106 mg mL-1, MA10: 1.345 mg mL-1 and MA05-2: 1.362 mg mL-1). Five strains (2SHXF01-3, MA03, MA05-2, MA01 and MA08-1) were selected based on their high cytotoxic activities. The ethyl acetate extract of the Nocardiopsis strain MA03 was particularly noted for its high antitumor activity against human carcinomas of the cervix (HeLa), lung (A549), kidney (Caki-1) and liver (HepG2) (IC50: 2.890, 1.981, 3.032 and 2.603 μg mL-1, respectively). The extract also remarkably inhibited colony formation of HeLa cells at an extremely low concentration (0.5 μg mL-1). This study highlights that marine-derived actinomycetes are a huge resource of compounds for the biological control of aflatoxin contamination and the development of novel drugs for human carcinomas.

  17. Geobacteraceae strains and methods

    Science.gov (United States)

    Lovley, Derek R.; Nevin, Kelly P.; Yi, Hana

    2015-07-07

    Embodiments of the present invention provide a method of producing genetically modified strains of electricigenic microbes that are specifically adapted for the production of electrical current in microbial fuel cells, as well as strains produced by such methods and fuel cells using such strains. In preferred embodiments, the present invention provides genetically modified strains of Geobacter sulfurreducens and methods of using such strains.

  18. Activated extracellular signal-regulated kinases are necessary and sufficient to initiate tubulogenesis in renal tubular MDCK strain I cell cysts.

    Science.gov (United States)

    Hellman, Nathan E; Greco, Andres J; Rogers, Katherine K; Kanchagar, Chitra; Balkovetz, Daniel F; Lipschutz, Joshua H

    2005-10-01

    A classic in vitro model of renal cyst and tubule formation utilizes the Madin-Darby canine kidney (MDCK) cell line, of which two strains exist. Most cyst and tubule formation studies that utilized MDCK cells have been performed with MDCK strain II cells. MDCK strain II cells form hollow cysts in a three-dimensional collagen matrix over 10 days and tubulate in response to hepatocyte growth factor, which increases levels of active (phosphorylated) ERK1/2. In this study, we demonstrate that MDCK strain I cells also form cysts when grown in a collagen matrix; however, MDCK strain I cell cysts spontaneously initiate the primary steps in tubulogenesis. Analysis of time-lapse microscopy of both MDCK strain I and strain II cell cysts during the initial stages of tubulogenesis demonstrates a highly dynamic process with cellular extensions and retractions occurring rapidly and continuously. MDCK strain I cell cysts can spontaneously initiate tubulogenesis mainly because of relatively higher levels of active ERK in MDCK strain I, compared with strain II, cells. The presence of either of two distinct inhibitors of ERK activation (UO126 and PD09059) prevents tubulogenesis from occurring spontaneously in MDCK strain I cell cysts and, in response to hepatocyte growth factor, in strain II cell cysts. The difference between MDCK strain I and strain II cell lines is likely explained by differing embryological origins, with strain I cells being of collecting duct, and hence ureteric bud, origin. Ureteric bud cells also have high levels of active ERK and spontaneously tubulate in our in vitro collagen gel system, with tubulogenesis inhibited by UO126 and PD09059. These results suggest that a seminal event in kidney development may be the activation of ERK in the mesonephric duct/ureteric bud cells destined to form the collecting tubules.

  19. Do mechanical strain and TNF-α interact to amplify pro-inflammatory cytokine production in human annulus fibrosus cells?

    Science.gov (United States)

    Likhitpanichkul, Morakot; Torre, Olivia M; Gruen, Jadry; Walter, Benjamin A; Hecht, Andrew C; Iatridis, James C

    2016-05-03

    During intervertebral disc (IVD) injury and degeneration, annulus fibrosus (AF) cells experience large mechanical strains in a pro-inflammatory milieu. We hypothesized that TNF-α, an initiator of IVD inflammation, modifies AF cell mechanobiology via cytoskeletal changes, and interacts with mechanical strain to enhance pro-inflammatory cytokine production. Human AF cells (N=5, Thompson grades 2-4) were stretched uniaxially on collagen-I coated chambers to 0%, 5% (physiological) or 15% (pathologic) strains at 0.5Hz for 24h under hypoxic conditions with or without TNF-α (10ng/mL). AF cells were treated with anti-TNF-α and anti-IL-6. ELISA assessed IL-1β, IL-6, and IL-8 production and immunocytochemistry measured F-actin, vinculin and α-tubulin in AF cells. TNF-α significantly increased AF cell pro-inflammatory cytokine production compared to basal conditions (IL-1β:2.0±1.4-84.0±77.3, IL-6:10.6±9.9-280.9±214.1, IL-8:23.9±26.0-5125.1±4170.8pg/ml for basal and TNF-α treatment, respectively) as expected, but mechanical strain did not. Pathologic strain in combination with TNF-α increased IL-1β, and IL-8 but not IL-6 production of AF cells. TNF-α treatment altered F-actin and α-tubulin in AF cells, suggestive of altered cytoskeletal stiffness. Anti-TNF-α (infliximab) significantly inhibited pro-inflammatory cytokine production while anti-IL-6 (atlizumab) did not. In conclusion, TNF-α altered AF cell mechanobiology with cytoskeletal remodeling that potentially sensitized AF cells to mechanical strain and increased TNF-α-induced pro-inflammatory cytokine production. Results suggest an interaction between TNF-α and mechanical strain and future mechanistic studies are required to validate these observations.

  20. Rickettsia massiliae and Rickettsia conorii Israeli Spotted Fever Strain Differentially Regulate Endothelial Cell Responses.

    Science.gov (United States)

    Bechelli, Jeremy; Smalley, Claire; Milhano, Natacha; Walker, David H; Fang, Rong

    2015-01-01

    Rickettsiae primarily target microvascular endothelial cells. However, it remains elusive how endothelial cell responses to rickettsiae play a role in the pathogenesis of rickettsial diseases. In the present study, we employed two rickettsial species with high sequence homology but differing virulence to investigate the pathological endothelial cell responses. Rickettsia massiliae is a newly documented human pathogen that causes a mild spotted fever rickettsiosis. The "Israeli spotted fever" strain of R. conorii (ISF) causes severe disease with a mortality rate up to 30% in hospitalized patients. At 48 hours post infection (HPI), R. conorii (ISF) induced a significant elevation of IL-8 and IL-6 while R. massiliae induced a statistically significant elevated amount of MCP-1 at both transcriptional and protein synthesis levels. Strikingly, R. conorii (ISF), but not R. massiliae, caused a significant level of cell death or injury in HMEC-1 cells at 72 HPI, demonstrated by live-dead cell staining, annexin V staining and lactate dehydrogenase release. Monolayers of endothelial cells infected with R. conorii (ISF) showed a statistically significant decrease in electrical resistance across the monolayer compared to both R. massiliae-infected and uninfected cells at 72 HPI, suggesting increased endothelial permeability. Interestingly, pharmacological inhibitors of caspase-1 significantly reduced the release of lactate dehydrogenase by R. conorii (ISF)-infected HMEC-1 cells, which suggests the role of caspase-1 in mediating the death of endothelial cells. Taken together, our data illustrated that a distinct proinflammatory cytokine profile and endothelial dysfunction, as evidenced by endothelial cell death/injury and increased permeability, are associated with the severity of rickettsial diseases.

  1. Variation of gravity before the Alxa Zuoqi M5.8 earthquake

    Directory of Open Access Journals (Sweden)

    Jianlin Feng

    2016-07-01

    Full Text Available In this study, a classic survey adjustment computation method was used for data obtained in the Inner Mongolia and Ningxia gravimetric networks between September 2013 and April 2015 so as to investigate the variation of gravity before the Alxa Zuoqi M5.8 earthquake. The relationship between gravity variation and the Alxa Zuoqi M5.8 earthquake was analyzed. The results showed that: (1 the severe variation in gravity field at the test sites before the Alxa Zuoqi M5.8 earthquake, as well as the subsequent accelerated rising, might be an earthquake precursor; (2 the Alxa Zuoqi M5.8 earthquake occurred at the turning point where the high-gravity gradient zone changed from the NE direction to NW.

  2. Atypical Enteropathogenic Escherichia coli Strains form Biofilm on Abiotic Surfaces Regardless of Their Adherence Pattern on Cultured Epithelial Cells

    Science.gov (United States)

    Culler, Hebert F.; Mota, Cristiane M.; Abe, Cecilia M.; Elias, Waldir P.; Sircili, Marcelo P.; Franzolin, Marcia R.

    2014-01-01

    The aim of this study was to determine the capacity of biofilm formation of atypical enteropathogenic Escherichia coli (aEPEC) strains on abiotic and biotic surfaces. Ninety-one aEPEC strains, isolated from feces of children with diarrhea, were analyzed by the crystal violet (CV) assay on an abiotic surface after 24 h of incubation. aEPEC strains representing each HEp-2 cell type of adherence were analyzed after 24 h and 6, 12, and 18 days of incubation at 37°C on abiotic and cell surfaces by CFU/cm2 counting and confocal laser scanning microscopy (CLSM). Biofilm formation on abiotic surfaces occurred in 55 (60.4%) of the aEPEC strains. There was no significant difference in biofilm biomass formation on an abiotic versus prefixed cell surface. The biofilms could be visualized by CLSM at various developmental stages. aEPEC strains are able to form biofilm on an abiotic surface with no association with their adherence pattern on HEp-2 cells with the exception of the strains expressing UND (undetermined adherence). This study revealed the capacity of adhesion and biofilm formation by aEPEC strains on abiotic and biotic surfaces, possibly playing a role in pathogenesis, mainly in cases of persistent diarrhea. PMID:24883330

  3. Atypical Enteropathogenic Escherichia coli Strains form Biofilm on Abiotic Surfaces Regardless of Their Adherence Pattern on Cultured Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Hebert F. Culler

    2014-01-01

    Full Text Available The aim of this study was to determine the capacity of biofilm formation of atypical enteropathogenic Escherichia coli (aEPEC strains on abiotic and biotic surfaces. Ninety-one aEPEC strains, isolated from feces of children with diarrhea, were analyzed by the crystal violet (CV assay on an abiotic surface after 24 h of incubation. aEPEC strains representing each HEp-2 cell type of adherence were analyzed after 24 h and 6, 12, and 18 days of incubation at 37°C on abiotic and cell surfaces by CFU/cm2 counting and confocal laser scanning microscopy (CLSM. Biofilm formation on abiotic surfaces occurred in 55 (60.4% of the aEPEC strains. There was no significant difference in biofilm biomass formation on an abiotic versus prefixed cell surface. The biofilms could be visualized by CLSM at various developmental stages. aEPEC strains are able to form biofilm on an abiotic surface with no association with their adherence pattern on HEp-2 cells with the exception of the strains expressing UND (undetermined adherence. This study revealed the capacity of adhesion and biofilm formation by aEPEC strains on abiotic and biotic surfaces, possibly playing a role in pathogenesis, mainly in cases of persistent diarrhea.

  4. Influence of anode potentials on selection of Geobacter strains in microbial electrolysis cells.

    Science.gov (United States)

    Commault, Audrey S; Lear, Gavin; Packer, Michael A; Weld, Richard J

    2013-07-01

    Through their ability to directly transfer electrons to electrodes, Geobacter sp. are key organisms for microbial fuel cell technology. This study presents a simple method to reproducibly select Geobacter-dominated anode biofilms from a mixed inoculum of bacteria using graphite electrodes initially poised at -0.25, -0.36 and -0.42 V vs. Ag/AgCl. The biofilms all produced maximum power density of approximately 270 m Wm(-2) (projected anode surface area). Analysis of 16S rRNA genes and intergenic spacer (ITS) sequences found that the biofilm communities were all dominated by bacteria closely related to Geobacter psychrophilus. Anodes initially poised at -0.25 V reproducibly selected biofilms that were dominated by a strain of G. psychrophilus that was genetically distinct from the strain that dominated the -0.36 and -0.42 V biofilms. This work demonstrates for the first time that closely related strains of Geobacter can have very different competitive advantages at different anode potentials.

  5. Uniaxial cyclic strain enhances adipose-derived stem cell fusion with skeletal myocytes

    Energy Technology Data Exchange (ETDEWEB)

    Andersen, Jens Isak; Juhl, Morten; Nielsen, Thøger; Emmersen, Jeppe; Fink, Trine; Zachar, Vladimir; Pennisi, Cristian Pablo, E-mail: cpennisi@hst.aau.dk

    2014-07-25

    Highlights: • Uniaxial cyclic tensile strain (CTS) applied to ASCs alone or in coculture with myogenic precursors. • CTS promoted the formation of a highly ordered array of parallel ASCs. • Without biochemical supplements, CTS did not support advanced myogenic differentiation of ASCs. • Mechanical stimulation of cocultures boosted fusion of ASCs with skeletal myoblasts. - Abstract: Although adult muscle tissue possesses an exceptional capacity for regeneration, in the case of large defects, the restoration to original state is not possible. A well-known source for the de novo regeneration is the adipose-derived stem cells (ASCs), which can be readily isolated and have been shown to have a broad differentiation and regenerative potential. In this work, we employed uniaxial cyclic tensile strain (CTS), to mechanically stimulate human ASCs to participate in the formation skeletal myotubes in an in vitro model of myogenesis. The application of CTS for 48 h resulted in the formation of a highly ordered array of parallel ASCs, but failed to support skeletal muscle terminal differentiation. When the same stimulation paradigm was applied to cocultures with mouse skeletal muscle myoblasts, the percentage of ASCs contributing to the formation of myotubes significantly exceeded the levels reported in the literature hitherto. In perspective, the mechanical strain may be used to increase the efficiency of incorporation of ASCs in the skeletal muscles, which could be found useful in diverse traumatic or pathologic scenarios.

  6. A Lactobacillus rhamnosus strain induces a heme oxygenase dependent increase in Foxp3+ regulatory T cells.

    Directory of Open Access Journals (Sweden)

    Khalil Karimi

    Full Text Available We investigated the consequences of feeding with a Lactobacillus species on the immune environment in GALT, and the role of dendritic cells and heme oxygenase-1 in mediating these responses. Feeding with a specific strain of Lactobacillus rhamnosus induced a significant increase in CD4+CD25+Foxp3+ functional regulatory T cells in GALT. This increase was greatest in the mesenteric lymph nodes and associated with a marked decrease in TNF and IFNγ production. Dendritic cell regulatory function and HO-1 expression was also increased. The increase in Foxp3+ T cells could be prevented by treatment with a heme oxygenase inhibitor. However, neither inhibition of heme oxygenase nor blockade of IL-10 and TGFβ prevented the inhibition of inflammatory cytokine production. In conclusion Lactobacillus feeding induced a tolerogenic environment in GALT. HO-1 was critical to the enhancement of Foxp3+ regulatory T cells while additional, as yet unknown, pathways were involved in the down-regulation of inflammatory cytokine production by T cells.

  7. A Francisella tularensis live vaccine strain that improves stimulation of antigen-presenting cells does not enhance vaccine efficacy.

    Directory of Open Access Journals (Sweden)

    Deanna M Schmitt

    Full Text Available Vaccination is a proven strategy to mitigate morbidity and mortality of infectious diseases. The methodology of identifying and testing new vaccine candidates could be improved with rational design and in vitro testing prior to animal experimentation. The tularemia vaccine, Francisella tularensis live vaccine strain (LVS, does not elicit complete protection against lethal challenge with a virulent type A Francisella strain. One factor that may contribute to this poor performance is limited stimulation of antigen-presenting cells. In this study, we examined whether the interaction of genetically modified LVS strains with human antigen-presenting cells correlated with effectiveness as tularemia vaccine candidates. Human dendritic cells infected with wild-type LVS secrete low levels of proinflammatory cytokines, fail to upregulate costimulatory molecules, and activate human T cells poorly in vitro. One LVS mutant, strain 13B47, stimulated higher levels of proinflammatory cytokines from dendritic cells and macrophages and increased costimulatory molecule expression on dendritic cells compared to wild type. Additionally, 13B47-infected dendritic cells activated T cells more efficiently than LVS-infected cells. A deletion allele of the same gene in LVS displayed similar in vitro characteristics, but vaccination with this strain did not improve survival after challenge with a virulent Francisella strain. In vivo, this mutant was attenuated for growth and did not stimulate T cell responses in the lung comparable to wild type. Therefore, stimulation of antigen-presenting cells in vitro was improved by genetic modification of LVS, but did not correlate with efficacy against challenge in vivo within this model system.

  8. Cell-associated hemolytic activity in environmental strains of Plesiomonas shigelloides expressing cell-free, iron-influenced extracellular hemolysin.

    Science.gov (United States)

    González-Rodríguez, Nieves; Santos, Jesús A; Otero, Andrés; García-López, María-Luisa

    2007-04-01

    Hemolysis is a means of providing pathogenic bacteria with heme iron in vivo. In a previous work, iron-influenced hemolytic activity against sheep erythrocytes was detected in cell-free supernatants, but not in the cell fraction of two environmental Plesiomonas shigelloides strains incubated without shaking. Both strains have the hugA gene, which encodes an outer membrane receptor required for heme iron utilization. The present study was undertaken to investigate the expression of a second hemolytic activity detected during aerated incubation in normal and iron-depleted tryptone soya broth (id-TSB). An agar overlay procedure and doubling dilution titrations were employed to detect the hemolytic activity against several erythrocyte species. The kinetics of growth and hemolytic activity were assayed at 35 degrees C in aerated normal and id-TSB and salmon extract. Overlaid colonies showed a cell-associated beta-hemolytic activity within 4 h. For aerated cell-free supernatants, titers above 16 were not attained until 30 to 48 h of incubation; the best activity was noted with dog and mouse erythrocytes. After 24 h of aerated incubation, sonicated cells yielded high hemolytic activity against dog erythrocytes without activity in supernatants, but after 48 h, only 28 to 30% of the total activity remained cell associated. The hemolytic factor was released in broths during the death phase. Hemolytic activity was not detected in fish extract. This and other studies suggest that P. shigelloides may produce at least two hemolytic factors, their expression and detection being influenced by environmental growth conditions and testing procedures. The overlay assay appears to be the best routine method for detecting hemolytic activity in P. shigelloides.

  9. Identification of novel genomic aberrations in AML-M5 in a level of array CGH.

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    Rui Zhang

    Full Text Available To assess the possible existence of unbalanced chromosomal abnormalities and delineate the characterization of copy number alterations (CNAs of acute myeloid leukemia-M5 (AML-M5, R-banding karyotype, oligonucelotide array CGH and FISH were performed in 24 patients with AML-M5. A total of 117 CNAs with size ranging from 0.004 to 146.263 Mb was recognized in 12 of 24 cases, involving all chromosomes other than chromosome 1, 4, X and Y. Cryptic CNAs with size less than 5 Mb accounted for 59.8% of all the CNAs. 12 recurrent chromosomal alterations were mapped. Seven out of them were described in the previous AML studies and five were new candidate AML-M5 associated CNAs, including gains of 3q26.2-qter and 13q31.3 as well as losses of 2q24.2, 8p12 and 14q32. Amplication of 3q26.2-qter was the sole large recurrent chromosomal anomaly and the pathogenic mechanism in AML-M5 was possibly different from the classical recurrent 3q21q26 abnormality in AML. As a tumor suppressor gene, FOXN3, was singled out from the small recurrent CNA of 14q32, however, it is proved that deletion of FOXN3 is a common marker of myeloid leukemia rather than a specific marker for AML-M5 subtype. Moreover, the concurrent amplication of MLL and deletion of CDKN2A were noted and it might be associated with AML-M5. The number of CNA did not show a significant association with clinico-biological parameters and CR number of the 22 patients received chemotherapy. This study provided the evidence that array CGH served as a complementary platform for routine cytogenetic analysis to identify those cryptic alterations in the patients with AML-M5. As a subtype of AML, AML-M5 carries both common recurrent CNAs and unique CNAs, which may harbor novel oncogenes or tumor suppressor genes. Clarifying the role of these genes will contribute to the understanding of leukemogenic network of AML-M5.

  10. Inactivation of a GAL4-Like Transcription Factor Improves Cell Fitness and Product Yield in Glycoengineered Pichia pastoris Strains

    Science.gov (United States)

    Argyros, Rebecca; Bukowski, John; Nelson, Stephanie; Sharkey, Nathan; Kim, Sehoon; Copeland, Victoria; Davidson, Robert C.; Chen, Ronghua; Zhuang, Jun; Sethuraman, Natarajan; Stadheim, Terrance A.

    2014-01-01

    With a completely reengineered and humanized glycosylation pathway, glycoengineered Pichia pastoris has emerged as a promising production host for the manufacture of therapeutic glycoproteins. However, the extensive genetic modifications have also negatively affected the overall fitness levels of the glycoengineered host cells. To make glycoengineered Pichia strains more compatible with a scalable industrial fermentation process, we sought to identify genetic solutions to broadly improve cell robustness during fermentation. In this study, we report that mutations within the Pichia pastoris ATT1 (PpATT1) gene (a homolog of the Saccharomyces cerevisiae GAL4 [ScGAL4] transcriptional activator) dramatically increased the cellular fitness levels of glycoengineered Pichia strains. We demonstrate that deletion of the PpATT1 gene enabled glycoengineered Pichia strains to improve their thermal tolerance levels, reduce their cell lysis defects, and greatly improve fermentation robustness. The extension of the duration of fermentation enabled the PpATT1-modified glycoengineered Pichia strains to increase their product yields significantly without any sacrifice in product quality. Because the ATT1 gene could be deleted from any Pichia strains, including empty hosts and protein-expressing production strains alike, we suggest that the findings described in this study are broadly applicable to any Pichia strains used for the production of therapeutic proteins, including monoclonal antibodies, Fc fusions, peptides, hormones, and growth factors. PMID:25344235

  11. Propagation of Brazilian Zika virus strains in static and suspension cultures using Vero and BHK cells.

    Science.gov (United States)

    Nikolay, Alexander; Castilho, Leda R; Reichl, Udo; Genzel, Yvonne

    2017-03-23

    The recent spread of Zika virus (ZIKV) in the Americas and the Pacific has reached alarming levels in more than 60 countries. However, relatively little is known about the disease on a virological and epidemiological level and its consequences for humans. Accordingly, a large demand for in vitro derived Brazilian ZIKV material to support in vitro and in vivo studies has arisen. However, a prompt supply of ZIKV and ZIKV antigens cannot be guaranteed as the production of this virus typically using Vero or C6/36 cell lines remains challenging. Here we present a production platform based on BHK-21 suspension (BHK-21SUS) cells to propagate Brazilian ZIKV at larger quantities in perfusion bioreactors. Scouting experiments performed in tissue culture flasks using adherent BHK-21 and Vero cells have demonstrated similar permissivity and virus yields for four different Brazilian ZIKV isolates. The cell-specific yield of infectious virus particles varied between respective virus strains (1-48PFU/cell), and the ZIKV isolate from the Brazilian state Pernambuco (ZIKV(PE)) showed to be a best performing isolate for both cell lines. However, infection studies of BHK-21SUS cells with ZIKV(PE) in shake flasks resulted in poor virus replication, with a maximum titer of 8.9×10(3)PFU/mL. Additional RT-qPCR measurements of intracellular and extracellular viral RNA levels revealed high viral copy numbers within the cell, but poor virus release. Subsequent cultivation in a perfusion bioreactor using an alternating tangential flow filtration system (ATF) under controlled process conditions enabled cell concentrations of about 1.2×10(7)cells/mL, and virus titers of 3.9×10(7)PFU/mL. However, while the total number of infectious virus particles was increased, the cell-specific yield (3.3PFU/cell) remained lower than determined in adherent cell lines. Nevertheless, the established perfusion process allows to provide large amounts of ZIKV material for research and is a first step towards

  12. Combined effects of surface morphology and mechanical straining magnitudes on the differentiation of mesenchymal stem cells without using biochemical reagents.

    Science.gov (United States)

    Jang, Ji-Yeon; Lee, Shi Woo; Park, So Hee; Shin, Ji Won; Mun, ChiWoong; Kim, Su-Hyang; Kim, Dong Hwa; Shin, Jung-Woog

    2011-01-01

    Existing studies examining the control of mesenchymal stem cell (MSC) differentiation into desired cell types have used a variety of biochemical reagents such as growth factors despite possible side effects. Recently, the roles of biomimetic microphysical environments have drawn much attention in this field. We studied MSC differentiation and changes in gene expression in relation to osteoblast-like cell and smooth muscle-like cell type resulting from various microphysical environments, including differing magnitudes of tensile strain and substrate geometries for 8 days. In addition, we also investigated the residual effects of those selected microphysical environment factors on the differentiation by ceasing those factors for 3 days. The results of this study showed the effects of the strain magnitudes and surface geometries. However, the genes which are related to the same cell type showed different responses depending on the changes in strain magnitude and surface geometry. Also, different responses were observed three days after the straining was stopped. These data confirm that controlling microenvironments so that they mimic those in vivo contributes to the differentiation of MSCs into specific cell types. And duration of straining engagement was also found to play important roles along with surface geometry.

  13. Combined Effects of Surface Morphology and Mechanical Straining Magnitudes on the Differentiation of Mesenchymal Stem Cells without Using Biochemical Reagents

    Directory of Open Access Journals (Sweden)

    Ji-Yeon Jang

    2011-01-01

    Full Text Available Existing studies examining the control of mesenchymal stem cell (MSC differentiation into desired cell types have used a variety of biochemical reagents such as growth factors despite possible side effects. Recently, the roles of biomimetic microphysical environments have drawn much attention in this field. We studied MSC differentiation and changes in gene expression in relation to osteoblast-like cell and smooth muscle-like cell type resulting from various microphysical environments, including differing magnitudes of tensile strain and substrate geometries for 8 days. In addition, we also investigated the residual effects of those selected microphysical environment factors on the differentiation by ceasing those factors for 3 days. The results of this study showed the effects of the strain magnitudes and surface geometries. However, the genes which are related to the same cell type showed different responses depending on the changes in strain magnitude and surface geometry. Also, different responses were observed three days after the straining was stopped. These data confirm that controlling microenvironments so that they mimic those in vivo contributes to the differentiation of MSCs into specific cell types. And duration of straining engagement was also found to play important roles along with surface geometry.

  14. Cell type specificity and mechanism of control of a gene may be reverted in different strains of Dictyostelium discoideum.

    Science.gov (United States)

    Mangiarotti, G; Giorda, R

    2000-06-21

    Twelve genes which are expressed exclusively in pre-spore cells of Dictyostelium strain AX3 are expressed exclusively in pre-stalk cells of strain AX2. One gene has the opposite behavior: it is expressed in pre-stalk cells in AX3 and in pre-spore cells in AX2. The change in cell type specificity involves a change in the mechanism of control of gene expression. When they are expressed in pre-stalk cells, genes are controlled at the level of transcription, whilst in pre-spore cells, they are controlled at the level of mRNA stability. Genes expressed in pre-stalk cells in strain AX2, fused with an AX2 pre-spore specific promoter, become regulated at the level of mRNA stability. These findings indicate that at least a group of pre-stalk mRNAs possess the cis-destabilizing element typical of pre-spore mRNAs, though they are not destabilized in disaggregated cells. This is due to the fact that ribosomal protein S6, phosphorylation of which is responsible for controlling the stability of pre-spore mRNAs, is not dephosphorylated in disaggregated pre-stalk cells. These cells lack an S6 phosphatase activity which has been purified from disaggregated pre-spore cells.

  15. Mapping the mechanome of live stem cells using a novel method to measure local strain fields in situ at the fluid-cell interface.

    Directory of Open Access Journals (Sweden)

    Min Jae Song

    Full Text Available During mesenchymal condensation, the initial step of skeletogenesis, transduction of minute mechanical forces to the nucleus is associated with up or down-regulation of genes, ultimately resulting in formation of the skeletal template and appropriate cell lineage commitment. The summation of these biophysical cues affects the cell's shape and fate. Here, we predict and measure surface strain, in live stem cells, in response to controlled delivery of stresses, providing a platform to direct short-term structure--function relationships and long-term fate decisions. We measure local strains on stem cell surfaces using fluorescent microbeads coated with Concanavalin A. During delivery of controlled mechanical stresses, 4-Dimensional (x,y,z,t displacements of the bound beads are measured as surface strains using confocal microscopy and image reconstruction. Similarly, micro-particle image velocimetry (μ-piv is used to track flow fields with fluorescent microspheres. The measured flow velocity gradient is used to calculate stress imparted by fluid drag at the surface of the cell. We compare strain measured on cell surfaces with those predicted computationally using parametric estimates of the cell's elastic and shear modulus. Finally, cross-correlating stress--strain data to measures of gene transcription marking lineage commitment enables us to create stress--strain--fate maps, for live stem cells in situ. The studies show significant correlations between live stem cell stress--strain relationships and lineage commitment. The method presented here provides a novel means to probe the live stem cell's mechanome, enabling mechanistic studies of the role of mechanics in lineage commitment as it unfolds.

  16. Human antigen-presenting cells respond differently to gut-derived probiotic bacteria but mediate similar strain-dependent NK and T cell activation

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen; Zeuthen, Louise Hjerrild; Ferlazzo, Guido

    2007-01-01

    , in vitro assessment of the immunomodulatory effects of distinct strains may depend strongly on the cell type used as a model. To select the most appropriate model for screening of beneficial bacteria in human cells, the response to strains of intestinal bacteria of three types of antigen-presenting cells......The intestinal microbiota is essential for homeostasis of the local and systemic immune system, and particularly strains of lactic acid bacteria and Escherichia coli have been shown to have balancing effects on inflammatory conditions such as allergy and inflammatory bowel disease. However...... (APC) was compared; blood myeloid dendritic cells (DC), monocyte-derived DC and monocytes, and the effector response of natural killer cells and naïve T cells was characterized. Maturation induced by gut-derived bacteria differed between APC, with blood DC and monocytes responding with the production...

  17. Human microvascular endothelial cell toxicity caused by Brazilian purpuric fever-associated strains of Haemophilus influenzae biogroup aegyptius.

    Science.gov (United States)

    Weyant, R S; Quinn, F D; Utt, E A; Worley, M; George, V G; Candal, F J; Ades, E W

    1994-02-01

    An in vitro cytotoxicity model that uses an immortalized human microvascular endothelial cell line (HMEC-1) differentiates Brazilian purpuric fever (BPF)-associated Haemophilus influenzae biogroup aegyptius (HAE) strains from non-BPF-associated HAE strains. Toxic strains produced a characteristic HMEC-1 phenotype at an MOI of 1000 bacteria/TCC to produce an observable effect. The cytotoxic phenotype was characterized by the presence of large clumps of HMEC-1 cells, which detached from the monolayer within 48 h of inoculation by HAE cells. The cytotoxic phenotype was observed with 100% of BPF-associated HAE (40/40) and 14% of non-BPF-associated HAE (8/57; P < .001). The ability to study a BPF-associated phenotype in vitro using human microvascular cells should enhance our knowledge of BPF pathogenesis.

  18. Suitability of PER.C6 cells to generate epidemic and pandemic influenza vaccine strains by reverse genetics

    NARCIS (Netherlands)

    Koudstaal, W.; Hartgroves, L.; Havenga, M.; Legastelois, I.; Ophorst, C.; Siewerts, M.; Zuijdgeest, D.; Vogels, R.; Custers, J.; Boer-Luijtze, E. de; Leeuw, O. de; Cornelissen, L.; Goudsmit, J.; Barclay, W.

    2009-01-01

    Reverse genetics, the generation of influenza viruses from cDNA, presents a rapid method for creating vaccine strains. The technique necessitates the use of cultured cells. Due to technical and regulatory requirements, the choice of cell lines for production of human influenza vaccines is limited. P

  19. Suitability of PER.C6 cells to generate epidemic and pandemic influenza vaccine strains by reverse genetics

    NARCIS (Netherlands)

    Koudstaal, W.; Hartgroves, L.; Havenga, M.; Legastelois, I.; Ophorst, C.; Siewerts, M.; Zuijdgeest, D.; Vogels, R.; Custers, J.; Boer-Luijtze, E. de; Leeuw, O. de; Cornelissen, L.; Goudsmit, J.; Barclay, W.

    2009-01-01

    Reverse genetics, the generation of influenza viruses from cDNA, presents a rapid method for creating vaccine strains. The technique necessitates the use of cultured cells. Due to technical and regulatory requirements, the choice of cell lines for production of human influenza vaccines is limited. P

  20. Structure and abrasive wear resistance of R6M5 steel-tungsten carbide composite coatings

    Science.gov (United States)

    Gnyusov, S. F.

    2012-09-01

    Features of the structure formation, composition, and abrasive wear resistance of R6M5 steel-tungsten carbide (R6M5-WC) composite coatings have been studied as dependent on the WC content. The introduction of ˜20 wt % WC into the hardening composition leads to an increase in the fraction of M6C carbide (in the form of eutectic inclusions with average size ˜5.9 μm at grain boundaries and dispersed ˜0.25 μm particles in the volume of grains), while a large proportion of metastable austenite (˜88 vol %) is still retained. The R6M5-WC coatings exhibit high abrasive wear resistance, which is ensured by the γ → α' martensite transformation during friction and a muiltimodal size distribution of hardening particles.

  1. Mechanical strain modulates age-related changes in the proliferation and differentiation of mouse adipose-derived stromal cells

    Directory of Open Access Journals (Sweden)

    Chiang Wen-Sheng

    2010-03-01

    Full Text Available Abstract Background Previous studies on the effects of aging in human and mouse mesenchymal stem cells suggest that a decline in the number and differentiation potential of stem cells may contribute to aging and aging-related diseases. In this report, we used stromal cells isolated from adipose tissue (ADSCs of young (8-10 weeks, adult (5 months, and old (21 months mice to test the hypothesis that mechanical loading modifies aging-related changes in the self-renewal and osteogenic and adipogenic differentiation potential of these cells. Results We show that aging significantly reduced the proliferation and increased the adipogenesis of ADSCs, while the osteogenic potential is not significantly reduced by aging. Mechanical loading (10% cyclic stretching, 0.5 Hz, 48 h increased the subsequent proliferation of ADSCs from mice of all ages. Although the number of osteogenic colonies with calcium deposition was increased in ADSCs subjected to pre-strain, it resulted from an increase in colony number rather than from an increase in osteogenic potential after strain. Pre-strain significantly reduced the number of oil droplets and the expression of adipogenic marker genes in adult and old ADSCs. Simultaneously subjecting ADSCs to mechanical loading and adipogenic induction resulted in a stronger inhibition of adipogenesis than that caused by pre-strain. The reduction of adipogenesis by mechanical strain was loading-magnitude dependent: loading with 2% strain only resulted in a partial inhibition, and loading with 0.5% strain could not inhibit adipogenesis in ADSCs. Conclusions We demonstrate that mechanical stretching counteracts the loss of self-renewal in aging ADSCs by enhancing their proliferation and, at the same time, reduces the heightened adipogenesis of old cells. These findings are important for the further study of stem cell control and treatment for a variety of aging related diseases.

  2. Cell envelope components influencing filament length in the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120.

    Science.gov (United States)

    Burnat, Mireia; Schleiff, Enrico; Flores, Enrique

    2014-12-01

    Heterocyst-forming cyanobacteria grow as chains of cells (known as trichomes or filaments) that can be hundreds of cells long. The filament consists of individual cells surrounded by a cytoplasmic membrane and peptidoglycan layers. The cells, however, share a continuous outer membrane, and septal proteins, such as SepJ, are important for cell-cell contact and filament formation. Here, we addressed a possible role of cell envelope components in filamentation, the process of producing and maintaining filaments, in the model cyanobacterium Anabaena sp. strain PCC 7120. We studied filament length and the response of the filaments to mechanical fragmentation in a number of strains with mutations in genes encoding cell envelope components. Previously published peptidoglycan- and outer membrane-related gene mutants and strains with mutations in two genes (all5045 and alr0718) encoding class B penicillin-binding proteins isolated in this work were used. Our results show that filament length is affected in most cell envelope mutants, but the filaments of alr5045 and alr2270 gene mutants were particularly fragmented. All5045 is a dd-transpeptidase involved in peptidoglycan elongation during cell growth, and Alr2270 is an enzyme involved in the biosynthesis of lipid A, a key component of lipopolysaccharide. These results indicate that both components of the cell envelope, the murein sacculus and the outer membrane, influence filamentation. As deduced from the filament fragmentation phenotypes of their mutants, however, none of these elements is as important for filamentation as the septal protein SepJ.

  3. Large scale validation of the M5L lung CAD on heterogeneous CT datasets

    Energy Technology Data Exchange (ETDEWEB)

    Lopez Torres, E., E-mail: Ernesto.Lopez.Torres@cern.ch, E-mail: cerello@to.infn.it [CEADEN, Havana 11300, Cuba and INFN, Sezione di Torino, Torino 10125 (Italy); Fiorina, E.; Pennazio, F.; Peroni, C. [Department of Physics, University of Torino, Torino 10125, Italy and INFN, Sezione di Torino, Torino 10125 (Italy); Saletta, M.; Cerello, P., E-mail: Ernesto.Lopez.Torres@cern.ch, E-mail: cerello@to.infn.it [INFN, Sezione di Torino, Torino 10125 (Italy); Camarlinghi, N.; Fantacci, M. E. [Department of Physics, University of Pisa, Pisa 56127, Italy and INFN, Sezione di Pisa, Pisa 56127 (Italy)

    2015-04-15

    Purpose: M5L, a fully automated computer-aided detection (CAD) system for the detection and segmentation of lung nodules in thoracic computed tomography (CT), is presented and validated on several image datasets. Methods: M5L is the combination of two independent subsystems, based on the Channeler Ant Model as a segmentation tool [lung channeler ant model (lungCAM)] and on the voxel-based neural approach. The lungCAM was upgraded with a scan equalization module and a new procedure to recover the nodules connected to other lung structures; its classification module, which makes use of a feed-forward neural network, is based of a small number of features (13), so as to minimize the risk of lacking generalization, which could be possible given the large difference between the size of the training and testing datasets, which contain 94 and 1019 CTs, respectively. The lungCAM (standalone) and M5L (combined) performance was extensively tested on 1043 CT scans from three independent datasets, including a detailed analysis of the full Lung Image Database Consortium/Image Database Resource Initiative database, which is not yet found in literature. Results: The lungCAM and M5L performance is consistent across the databases, with a sensitivity of about 70% and 80%, respectively, at eight false positive findings per scan, despite the variable annotation criteria and acquisition and reconstruction conditions. A reduced sensitivity is found for subtle nodules and ground glass opacities (GGO) structures. A comparison with other CAD systems is also presented. Conclusions: The M5L performance on a large and heterogeneous dataset is stable and satisfactory, although the development of a dedicated module for GGOs detection could further improve it, as well as an iterative optimization of the training procedure. The main aim of the present study was accomplished: M5L results do not deteriorate when increasing the dataset size, making it a candidate for supporting radiologists on large

  4. Differential effects of acute morphine administrations on polymorphonuclear cell metabolism in various mouse strains.

    Science.gov (United States)

    Di Francesco, P; Tavazzi, B; Gaziano, R; Lazzarino, G; Casalinuovo, I A; Di Pierro, D; Garaci, E

    1998-01-01

    This paper shows that an acute morphine treatment dose-dependently alters the energetic and oxidative metabolism of polymorphonuclear leukocytes obtained from BALB/c and DBA/2 mice, while phagocytic cells from C57BL/6 were not affected. In sensitive mouse strains, i.e. BALB/c and DBA/2, morphine decreased both ATP concentration and energy charge potential. At the same time, ATP catabolic products, i.e. nucleosides (inosine+adenosine) and oxypurines (hypoxanthine+xanthine+uric acid), significantly increased, indicating an imbalance between energy production and consumption. Morphine treatment also induced malondialdehyde and superoxide anions production in leukocyte cells from sensitive mice. The opiate antagonist naloxone blocked morphine-induced modifications by the lower morphine dose. The same parameters in cells from C57BL/6 mice were not affected. These findings confirm that: i) the phagocytic cells are an important target for the in vivo effects of morphine, and ii) the genotype-dependent variation influences the immunological responsiveness to opiates.

  5. Pulmonary neuroendocrine cell hyperplasia: identification, diagnostic criteria and incidence in untreated ageing rats of different strains.

    Science.gov (United States)

    Haworth, Richard; Woodfine, Jennie; McCawley, Sean; Pilling, Andrew M; Lewis, David J; Williams, Tom C

    2007-08-01

    Pulmonary Neuroendocrine Cells (PNEC) are found as clusters called neuroepithelial bodies (NEB) or as single cells scattered in the respiratory epithelium. Pulmonary neuroendocrine cell hyperplasia is recorded in humans and experimentally manipulated rodents. The objectives of this work were to identify the optimal immunohistochemical markers for PNEC in the rat for use on paraffin-embedded, formalin-fixed material and to provide the first comparative incidence of PNEC hyperplasia in untreated 2-year-old rats of different strains. Calcitonin-gene related peptide (CGRP) and protein G product 9.5 (PGP9.5) antibodies identified PNEC consistently and selectively. In contrast, PNEC did not express chromogranin-A or S-100. PNEC hyperplasia was defined as foci of PNEC with greater than 40 nuclei, excluding overlying respiratory epithelium and submucosal PNEC. PNEC hyperplasia was observed at low incidence (0-7%) in untreated 2-year-old Sprague-Dawley, Han Wistar and Wistar rats but not Fischer 344 rats. This is the first report of spontaneous PNEC hyperplasia in rats. The cause of this hyperplasia is unknown, but experimental models that induce PNEC hyperplasia by causing bronchiolar cell injury are discussed. PNEC neoplasia in the rat is unreported in the literature and was not observed in animals examined in this study.

  6. Miltefosine-induced apoptotic cell death on Leishmania major and L. tropica strains.

    Science.gov (United States)

    Khademvatan, Shahram; Gharavi, Mohammad Javad; Rahim, Fakher; Saki, Jasem

    2011-03-01

    The aim of this study was to assess the cytotoxic effects of various concentrations of miltefosine on Leishmania major (MRHO/IR/75/ER) and L. tropica (MHOM/IR/02/Mash10) promastigotes and to observe the programmed cell death features. The colorimetric MTT assay was used to find L. major and L. tropica viability and the obtained results were expressed as 50% inhibitory concentration (IC50). Also, 50% effective doses (ED50) for L. major and L. tropica amastigotes were also determined. Annexin-V FLUOS staining was performed to study the cell death properties of miltefosine using FACS analysis. Qualitative analysis of the total genomic DNA fragmentation was performed by agarose gel electrophoresis. Furthermore, to observe changes in cell morphology, promastigotes were examined using light microscopy. In both strains of L. major and L. tropica, miltefosine induced dose-dependent death with features of apoptosis, including cell shrinkage, DNA laddering, and externalization of phosphatidylserine. The IC50 was achieved at 22 µM and 11 µM for L. major and L. tropica after 48 hr of incubation, respectively. ED50 of L. major and L. tropica amastigotes were 5.7 µM and 4.2 µM, respectively. Our results indicate that miltefosine induces apoptosis of the causative agent of cutaneous leishmaniasis in a dose-dependent manner. Interestingly, L. major did not display any apoptotic changes when it was exposed to miltefosine in concentrations sufficient to kill L. tropica.

  7. Molecular profile of clonal strains of human skeletal stem/progenitor cells with different potencies

    Directory of Open Access Journals (Sweden)

    Brian J. Sworder

    2015-05-01

    Full Text Available Bone marrow stromal cells (BMSCs, also known as bone marrow-derived mesenchymal stem cells are fibroblastic reticular cells, a subset of which is composed of multipotent skeletal stem cells (SSCs. SSCs/BMSCs are able to recreate a bone/marrow organ in vivo. To determine differences between clonogenic multipotent SSCs and similarly clonogenic but non-multipotent BMSCs, we established single colony-derived strains (SCDSs, initiated by individual Colony Forming Unit-Fibroblasts and determined their differentiation capacity by vivo transplantation. In this series of human SCDSs (N = 24, 20.8% formed fibrous tissue (F, 66.7% formed bone (B, and 12.5% formed a bone/marrow organ, and thus were multipotent (M. RNA isolated from 12 SCDSs just prior to transplantation was analyzed by microarray. Although highly similar, there was variability from one SCDS to another, and SCDSs did not strictly segregate into the three functional groups (F, B or M by unsupervised hierarchical clustering. We then compared 3 F-SCDSs to 3 M-SCDSs that did segregate. Genes associated with skeletogenesis, osteoblastogeneis, hematopoiesis, and extracellular matrix were over-represented in M-SCDSs compared with F-SCDSs. These results highlight the heterogeneity of SSCs/BMSCs, even between functionally similar SCDSs, but also indicate that differences can be detected that may shed light on the character of the SSC.

  8. Expression of cyclooxygenase-2 mRNA in drug-sensitive cell and drug-resistant strains of ovarian cancer cell lines

    Institute of Scientific and Technical Information of China (English)

    Xiaoyan Li; Zehua Wang

    2006-01-01

    Objective: To investigate the expression of cyclooxygenase-2 (COX-2) mRNA in drug-sensitive cell and drugresistant clones of ovarian cancer cell lines. Methods: RT-PCR and immunocytochemistry were used to investigate the expression of cyclooxygenase-2 in 3 clones drug-sensitive and 5 clones drug-resistant ovarian cancer cell. Results: Strong COX-2 mRNA expressions were detected in 3 clones of drug-sensitive cell and weak expressions were detected in 5 clones of drug-resistant cell. The protein expression of COX-2 in drug-sensitive cell was strongly positive reaction in immunocytochemistry stain and there was a weak positive reaction in 5clones of drug-resistant cell. Conclusion: The expression of COX-2 mRNA in drug-sensitive cell strains is much higher than that in drugresistant strains of ovarian cancer cell lines, providing a basis of the chemoprevention for ovarian cancer.

  9. Comparing M5 Model Trees and Neural Networks for River Level Forecasting

    Science.gov (United States)

    Khan, S.; See, L.

    2005-12-01

    Artificial neural networks (ANNs) have been the subject of much research activity in hydrological modelling over the last decade yet this represents only one data-driven modelling approach from among a very rich set. M5 model trees are an example of a technique that has had little application in the hydrological domain yet the results are promising (Solomatine and Xue, 2004). They are a machine learning approach that combines regression trees and classification. The input space is partitioned into subsets based on entropy measures, and regression equations are then fit to these subsets. The advantages over ANNs are (a) their ability to provide knowledge in the form of a decision tree and (b) much faster training times. This has important implications for operational use as they are not black box models. In this study ANNs, M5 model trees and time series analysis have been used to develop models to predict river levels at a gauging station in the River Ouse catchment in Northern England. Two lead times have been used: t+6 and t+24 hours. The input data consisted of historical levels at the gauging stations, upstream level data and rainfall from five rain gauges across the catchment, determined by correlation with the output. The results of the study showed that the ANNs outperformed both the M5 model trees and time series approaches when considering global goodness-of-fit measures such as root mean squared error and coefficient of efficiency. However, the difference in performance between the ANNs and M5 model trees was not large, e.g. 1 percent difference in coefficient of efficiency for t+6 hours. When considering the longer lead time of t+24 hours, the performance of the ANNs and M5 model trees almost converged. The M5 model tree, however, also provides the rules of operation. The first partition for both the t+6 and t+24 hour models was determined by the value of the river level at one of the upstream stations. The individual regression equations associated with

  10. Hydrogen production by Clostridium acetobutylicum ATCC 824 and megaplasmid-deficient mutant M5 evaluated using a large headspace volume technique

    Energy Technology Data Exchange (ETDEWEB)

    Oh, Sang-Eun [Department of Biological Environment, Kangwon National University, Kangwon-do (Korea); Zuo, Yi; Zhang, Husen [Department of Civil and Environmental Engineering, The Pennsylvania State University, University Park, PA 16802 (United States); Guiltinan, Mark J. [Department of Horticulture, The Pennsylvania State University, University Park, PA 16802 (United States); Hydrogen Energy (H2E) Center, The Pennsylvania State University, University Park, PA 16802 (United States); Logan, Bruce E.; Regan, John M. [Department of Civil and Environmental Engineering, The Pennsylvania State University, University Park, PA 16802 (United States); Hydrogen Energy (H2E) Center, The Pennsylvania State University, University Park, PA 16802 (United States)

    2009-12-15

    Biohydrogen production is measured using a variety of techniques, ranging from low cost intermittent gas release methods where yields are usually reduced due to high partial pressures of hydrogen, to expensive respirometers that can eliminate pressure buildup. A new large headspace volume technique was developed that reduces the potential for hydrogen gas inhibition without the need for a respirometer. We tested this method with two strains of clostridia, Clostridium acetobutylicum ATCC 824 and its mutant M5 that lacks a megaplasmid responsible for butanol and acetone production, and a mixed culture (heat-treated sludge). The hydrogen yield using M5 (2.64 mol-H{sub 2}/mol-glucose) was 47% higher than that of the parent strain (1.79 mol-H{sub 2}/mol-glucose), and 118% larger than that obtained in tests with the sludge inoculum (1.21 mol-H{sub 2}/mol-glucose). The increased yield for M5 was primarily due to a decrease in biomass synthesis (38%) compared to the parent strain. Hydrogen yields measured using this new method were on average 14% higher than those obtained using a conventional respirometric method. These findings indicate enhanced biohydrogen production from the megaplasmid-deficient mutant of C. acetobutylicum ATCC 824, and that an intermittent gas-sampling technique can effectively measure high hydrogen gas by using a large headspace volume. (author)

  11. Biologic characteristic studies of DNA mismatch—repair enzyme hMSH2—deficient cell strain

    Institute of Scientific and Technical Information of China (English)

    HeY; ZhuaZX

    2002-01-01

    The effect of hMSH2 enzyme-deficiency on the cell growing phenotypes,cell ultrastructure,growth character and cell cycle were observed with electronic microscopy examination,cell counting and flow cytometry.hMSH2-deficient cell strain was constructed by transfecting hMSH2 recombination plasmid of antisense RNA into human embryo lung fibroblasts(HLF).In hMSH2-deficient cells,there were a lot of morphological changes under electronic microscopy,such as irregular shape,a lot of protuberances on the surface of cell,the enlarged nuclei.The average time of double increment of HLF and hMSH2-deficient cells were 1.0d and 0.78d,respectively.This suggested that the cell proliferation of hMSH2-deficient cells was greater than that of HLF.The distribution of HLF and hMSH2-deficient cells in G1,G2 and S phases was different.A large part of hMSH2-deficient cells was blocked in G1 phase.hMSH2-deficient cells increased,but it is still not a typical malignant cells.Thus,this cell strain could be used as biologic material to detect mutagenesis of environmental chemicals.

  12. Cytopathic effects of toxogenic strains of Helicobacter pylori on different cell lines

    Directory of Open Access Journals (Sweden)

    K. Lakshmana Gowda

    2014-01-01

    Full Text Available Purpose: Many virulence factors are involved in the pathomechanism of infection caused by Helicobacter pylori. Toxins such as vacuolating cytotoxin, encoded by the vacA gene and the immunogenic protein cagA, encoded by the cagA gene (cytotoxin-associated gene are major factors conferring the property of virulence. The current study is aimed at isolation of H. pylori and separation of its toxin from antral biopsies of patients. Materials and Methods: The following cell lines were used to demonstrate the cytopathic effect (CPE of the separated toxin: African green monkey kidney (Vero, baby hamster kidney, human lung carcinoma (LLC-MK2, and human epithelial. Results: H. pylori was isolated from 27 out of 45 patients (60% selected for the study. CPE of H. pylori toxin was highly significant on Vero cells than other cell lines used as it reached a high dilution titer of toxin (1/16 in 13 isolated strains (48.15%. No significant difference in CPE of toxin in different dilutions was detected among other cell lines used in different groups. H. pylori toxin could be detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis as a distinct band with a molecular weight ranging between 66 and 97 kDa and closely related to 87 kDa. Conclusion: H. pylori vacuolating cytotoxin plays a vital role in the pathogenesis of gastroduodenal diseases (gastritis, gastric ulcer, duodenal ulcer, and gastric cancer. The Vero cell lines were found to be the most suitable form of tissue culture when compared with other cell lines used in our study for demonstrating the activity of H. pylori toxin.

  13. Immunoregulatory effect of bifidobacteria strains in porcine intestinal epithelial cells through modulation of ubiquitin-editing enzyme A20 expression.

    Directory of Open Access Journals (Sweden)

    Yohsuke Tomosada

    Full Text Available BACKGROUND: We previously showed that evaluation of anti-inflammatory activities of lactic acid bacteria in porcine intestinal epithelial (PIE cells is useful for selecting potentially immunobiotic strains. OBJECTIVE: The aims of the present study were: i to select potentially immunomodulatory bifidobacteria that beneficially modulate the Toll-like receptor (TLR-4-triggered inflammatory response in PIE cells and; ii to gain insight into the molecular mechanisms involved in the anti-inflammatory effect of immunobiotics by evaluating the role of TLR2 and TLR negative regulators in the modulation of proinflammatory cytokine production and activation of mitogen-activated protein kinase (MAPK and nuclear factor-κB (NF-κB pathways in PIE cells. RESULTS: Bifidobacteria longum BB536 and B. breve M-16V strains significantly downregulated levels of interleukin (IL-8, monocyte chemotactic protein (MCP-1 and IL-6 in PIE cells challenged with heat-killed enterotoxigenic Escherichia coli. Moreover, BB536 and M-16V strains attenuated the proinflammatory response by modulating the NF-κB and MAPK pathways. In addition, our findings provide evidence for a key role for the ubiquitin-editing enzyme A20 in the anti-inflammatory effect of immunobiotic bifidobacteria in PIE cells. CONCLUSIONS: We show new data regarding the mechanism involved in the anti-inflammatory effect of immunobiotics. Several strains with immunoregulatory capabilities used a common mechanism to induce tolerance in PIE cells. Immunoregulatory strains interacted with TLR2, upregulated the expression of A20 in PIE cells, and beneficially modulated the subsequent TLR4 activation by reducing the activation of MAPK and NF-κB pathways and the production of proinflammatory cytokines. We also show that the combination of TLR2 activation and A20 induction can be used as biomarkers to screen and select potential immunoregulatory bifidobacteria strains.

  14. Anisotropic emission and photon-recycling in strain-balanced quantum well solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Cabrera, C. I.; Enciso, A.; Contreras-Solorio, D. A. [Academic Unit of Physics, Autonomous University of Zacatecas, Czda. Solidaridad y Paseo La Bufa S/N, 98060 Zacatecas, Zac. (Mexico); Rimada, J. C. [Solar Cell Laboratory, Institute of Materials Science and Technology (IMRE), University of Havana, Zapata y G, 10400 La Habana (Cuba); Hernandez, L., E-mail: luisman@fisica.uh.cu [Faculty of Physics, University of Havana, Colina Universitaria. 10400 La Habana (Cuba); Connolly, J. P. [Nanophotonics Technology Center, Universidad Politécnica de Valencia, 46022 Valencia (Spain)

    2014-04-28

    Strain-balanced quantum well solar cells (SB-QWSCs) extend the photon absorption edge beyond that of bulk GaAs by incorporation of quantum wells in the i-region of a p–i–n device. Anisotropy arises from a splitting of the valence band due to compressive strain in the quantum wells, suppressing a transition which contributes to emission from the edge of the quantum wells. We have studied both the emission light polarized in the plane perpendicular (TM) to the quantum well which couples exclusively to the light hole transition and the emission polarized in the plane of the quantum wells (TE) which couples mainly to the heavy hole transition. It was found that the spontaneous emission rates TM and TE increase when the quantum wells are deeper. The addition of a distributed Bragg reflector can substantially increase the photocurrent while decreasing the radiative recombination current. We have examined the impact of the photon recycling effect on SB-QWSC performance. We have optimized SB-QWSC design to achieve single junction efficiencies above 30%.

  15. Cytoskeletal Strains in Modeled Optohydrodynamically Stressed Healthy and Diseased Biological Cells

    Directory of Open Access Journals (Sweden)

    Sean S. Kohles

    2012-01-01

    Full Text Available Controlled external chemomechanical stimuli have been shown to influence cellular and tissue regeneration/degeneration, especially with regards to distinct disease sequelae or health maintenance. Recently, a unique three-dimensional stress state was mathematically derived to describe the experimental stresses applied to isolated living cells suspended in an optohydrodynamic trap (optical tweezers combined with microfluidics. These formulae were previously developed in two and three dimensions from the fundamental equations describing creeping flows past a suspended sphere. The objective of the current study is to determine the full-field cellular strain response due to the applied three-dimensional stress environment through a multiphysics computational simulation. In this investigation, the multiscale cytoskeletal structures are modeled as homogeneous, isotropic, and linearly elastic. The resulting computational biophysics can be directly compared with experimental strain measurements, other modeling interpretations of cellular mechanics including the liquid drop theory, and biokinetic models of biomolecule dynamics. The described multiphysics computational framework will facilitate more realistic cytoskeletal model interpretations, whose intracellular structures can be distinctly defined, including the cellular membrane substructures, nucleus, and organelles.

  16. Hidden selection rules, M5-instantons and fluxes in F-theory

    Energy Technology Data Exchange (ETDEWEB)

    Martucci, Luca [Dipartimento di Fisica e Astronomia ‘Galileo Galilei’, Università di Padova, & I.N.F.N. Sezione di Padova, via Marzolo 8, I-35131 Padova (Italy); Weigand, Timo [Institut für Theoretische Physik, Ruprecht-Karls-Universität, Philosophenweg 19, 69120 Heidelberg (Germany)

    2015-10-21

    We introduce a new approach to investigate the selection rules governing the contributions of fluxed M5-instantons to the F-theory four-dimensional effective action, with emphasis on the generation of charged matter F-terms. The structure of such couplings is unraveled by exploiting the perturbative and non-perturbative homological relations, introduced in our companion paper http://dx.doi.org/10.1007/JHEP09(2015)198, which encode the interplay between the self-dual 3-form flux on the M5-brane, the background 4-form flux and certain fibral curves. The latter are wrapped by time-like M2-branes representing matter insertions in the instanton path integral. In particular, we clarify how fluxed M5-instantons detect the presence of geometrically massive U(1)s which are responsible for ‘hidden’ selection rules. We discuss how for non-generic embeddings the M5-instanton can probe ‘locally massless’ U(1) symmetries if the rank of its Mordell-Weil group is enhanced compared to that of the bulk. As a phenomenological off-spring we propose a new type of non-perturbative corrections to Yukawa couplings which may change the rank of the Yukawa matrix. Along the way, we also gain new insights into the structure of massive U(1) gauge fluxes in the stable degeneration limit.

  17. Plant Cell Protolytic Enzymes Activity under Exposure to Lectins of Endophytic and Epiphytic Azospirillum Strains

    Directory of Open Access Journals (Sweden)

    S.A. Alen’kina

    2016-05-01

    Full Text Available We studied the ability of lectins isolated from the surface of the two strains of nitrogen-fixing soil bacteria of the genus Azospirillum, A. brasilense Sp7 (epiphytic and A. brasilense Sp245 (endophytic, to show have a regulating effect on the activity of pectinolytic enzymes in the roots of wheat seedlings. Research results showed that the lectins under study can cause the induction of the activity of polygalacturonase, pectinesterase, pectatlyase from the plant cell wall, thereby ensuring the bacteria penetration in the plant tissues, as well as the induction of plants responses which, being combined with growth-stimulating effect of bacteria, contributes to the formation of plants stability and productivity.

  18. The November 2011 M5.7 Oklahoma Earthquake: Induced or Triggered?

    Science.gov (United States)

    Sumy, D. F.; Cochran, E. S.; Aminzadeh, F.

    2013-12-01

    On 6 November 2011, a M5.7 right-lateral strike-slip earthquake ruptured a section of the Wilzetta fault that strikes approximately N55E in Prague, Oklahoma. This earthquake was preceded by a right-lateral strike-slip M5.0 foreshock that occurred on 5 November 2011 and followed by a left-lateral strike-slip M5.0 aftershock that occurred on 8 November 2011. Seismicity during the sequence delineates three distinct near-vertical fault planes. The M5.0 foreshock was within several hundred meters of two active, high-volume injection wells, and thus was interpreted as potentially induced by Keranen et al. [2013]. Immediately following the M5.0 foreshock, three temporary seismometers were installed, with additional 44 stations installed after the M5.7 mainshock. These 47 stations recorded thousands of foreshocks and aftershocks that surrounded the three M≥5.0 events. We hypothesize that while the M5.0 foreshock (Event A) is potentially induced by anthropogenic activities, the static stress change imparted by this event triggered the M5.7 mainshock (Event B). To investigate this hypothesis, we calculate the static Coulomb stress change on 110 focal mechanism solutions examined in this study. Many of the focal mechanisms are consistent with the rupture planes defined by the seismicity of Events A and B. However, several of the focal mechanism solutions exhibit dip-slip focal mechanisms and/or are more consistent with the M5.0 aftershock (Event C). Event C occurs on a previously unmapped, east-west striking left-lateral fault plane, which differs substantially from the orientation of Events A and B. The diverse range of focal mechanism solutions suggests complex fault interactions within the Wilzetta fault system. Based on these focal mechanism solutions, we investigate the static stress changes imparted on the aftershocks that result from each of the M≥5.0 earthquakes. We find that the stress change induced by the Event A increases the stress at the hypocentral location

  19. Effects of the Probiotic Enterococcus faecium and Pathogenic Escherichia coli Strains in a Pig and Human Epithelial Intestinal Cell Model

    Directory of Open Access Journals (Sweden)

    Ulrike Lodemann

    2015-01-01

    Full Text Available The aim of this study has been to elucidate the effect of the probiotic Enterococcus faecium NCIMB 10415 on epithelial integrity in intestinal epithelial cells and whether pre- and coincubation with this strain can reproducibly prevent damage induced by enterotoxigenic (ETEC and enteropathogenic Escherichia coli (EPEC. Porcine (IPEC-J2 and human (Caco-2 intestinal epithelial cells were incubated with bacterial strains and epithelial integrity was assessed by measuring transepithelial electrical resistance (TEER and mannitol flux rates. E. faecium alone increased TEER of Caco-2 cells without affecting mannitol fluxes whereas the E. coli strains decreased TEER and concomitantly increased mannitol flux rates in both cell lines. Preincubation with E. faecium had no effect on the TEER decrease induced by E. coli in preliminary experiments. However, in a second set of experiments using a slightly different protocol, E. faecium ameliorated the TEER decrease induced by ETEC at 4 h in IPEC-J2 and at 2, 4, and 6 h in Caco-2 cells. We conclude that E. faecium positively affected epithelial integrity in monoinfected Caco-2 cells and could ameliorate the damage on TEER induced by an ETEC strain. Reproducibility of the results is, however, limited when experiments are performed with living bacteria over longer periods.

  20. Quantification and characterization of cell wall polysaccharides released by non-Saccharomyces yeast strains during alcoholic fermentation.

    Science.gov (United States)

    Giovani, Giovanna; Rosi, Iolanda; Bertuccioli, Mario

    2012-11-15

    In order to improve knowledge about the oenological characteristics of non-Saccharomyces yeast strains, and to reconsider their contribution to wine quality, we studied the release of polysaccharides by 13 non-Saccharomyces strains of different species (three wine yeasts, six grape yeasts, and three spoilage yeasts) during alcoholic fermentation in synthetic must. Three Saccharomyces cerevisiae strains were included for comparison. All of the non-Saccharomyces strains released polysaccharides into fermentation medium; the amount released depended on the yeast species, the number of cells formed and their physiological conditions. Normalizing the quantity of macromolecules released to the cell biomass revealed that most non-Saccharomyces strains produced a greater quantity of polysaccharides compared to S. cerevisiae strains after 7 and 14days of fermentation. This capacity was particularly expressed in the studied wine spoilage yeasts (Saccharomycodes ludwigii, Zygosaccharomyces bailii, and Brettanomyces bruxellensis). Chemical characterization of exocellular polysaccharides produced by non-Saccharomyces yeasts revealed them to essentially be mannoproteins with high mannose contents, ranging from 93% for S'codes. ludwigii to 73-74% for Pichia anomala and Starmerella bombicola. Protein contents varied from 9% for P. anomala to 29% for Z. bailii. These compositions were very similar to those of the S. cerevisiae strains, and to the chemical composition of the cell wall mannoproteins of different yeast species. The presence of galactose, in addition to mannose and glucose, in the exocellular polysaccharides released by Schizosaccharomyces pombe, confirmed the parietal nature of the polysaccharides released by non-Saccharomyces yeasts; only this species has a galactomannan located in the outer layer of the cell wall.

  1. Influence of gastrointestinal system conditions on adhesion of exopolysaccharide-producing Lactobacillus delbrueckii subsp. bulgaricus strains to caco-2 cells

    Directory of Open Access Journals (Sweden)

    Derya Onal Darilmaz

    2011-10-01

    Full Text Available This study aimed to assess the transit tolerance of potential probiotic dairy Lactobacillus strains in human uppergastrointestinal tract in vitro, and to evaluate the effect of EPS production on the viability and adhesion of these strains. Survival and adhesion of two exopolysaccharide (EPS-producing L. delbrueckii subsp. bulgaricus strains (B3 and B2 and E. coli ATCC11229 were assessed after the exposure of different pH (gastric juice and gastric plus pancreatic juice challenges. In the artificial gastric juice (pH 2, both the viability of the strain B3 and B2 was decreased. Artificial juice treatments significantly reduced the adhesion to caco-2 cells (P< 0.05. High EPS-producing B3 survived better in the adverse gastrointestinal conditions and showed better ability of adhesion to Caco-2 cells when assessed for competition with E. coli ATCC 11229 compared to low EPS-producing B2. This investigation showed that EPS production could be affected or be involved in the viability, adherence and competition of L. delbrueckii subsp. bulgaricus strains and support the potential of B3 strain for development of new probiotic products.

  2. Human antigen-presenting cells respond differently to gut-derived probiotic bacteria but mediate similar strain-dependent NK and T cell activation.

    Science.gov (United States)

    Fink, Lisbeth N; Zeuthen, Louise H; Ferlazzo, Guido; Frøkiaer, Hanne

    2007-12-01

    The intestinal microbiota is essential for homeostasis of the local and systemic immune system, and particularly strains of lactic acid bacteria and Escherichia coli have been shown to have balancing effects on inflammatory conditions such as allergy and inflammatory bowel disease. However, in vitro assessment of the immunomodulatory effects of distinct strains may depend strongly on the cell type used as a model. To select the most appropriate model for screening of beneficial bacteria in human cells, the response to strains of intestinal bacteria of three types of antigen-presenting cells (APC) was compared; blood myeloid dendritic cells (DC), monocyte-derived DC and monocytes, and the effector response of natural killer cells and naïve T cells was characterized. Maturation induced by gut-derived bacteria differed between APC, with blood DC and monocytes responding with the production of IL-6 and tumour necrosis factor-alpha to bacteria, which elicited mainly IL-10 in monocyte-derived DC. In contrast, comparable IFN-gamma production patterns were found in both natural killer cells and T cells induced by all bacteria-matured APC. An inhibitory effect of certain strains on this IFN-gamma production was also mediated by all types of APC. The most potent responses were induced by monocyte-derived DC, which thus constitute a sensitive screening model.

  3. Study on persistent infection of Japanese encephalitis virus Beijing-1 strain in serum-free Sf9 cell cultures.

    Science.gov (United States)

    Kim, Hun; Lee, Su Jeen; Park, Jin Yong; Park, Yong Wook; Kim, Hyun Sung; Kang, Heui-Yun; Hur, Byung-Ki; Ryu, Yeon-Woo; Han, Sang In; Kim, Jong Su

    2004-03-01

    Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8 x 10(6) cells/ml in a 500ml spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-1 strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15% of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-1 strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-1 stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-1 strain rose from 1.0 x 10(5) to 1.5 x 10(6) pfu/ml. The infected Sf9 cells could be sub-cultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-1 strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-1 strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine

  4. ULE design considerations for a 3m class light weighted mirror blank for E-ELT M5

    Science.gov (United States)

    Fox, Andrew; Hobbs, Tom; Edwards, Mary; Arnold, Matthew; Sawyer, Kent

    2016-07-01

    It is expected that the next generation of large ground based astronomical telescopes will need large fast-steering/tip-tilt mirrors made of ultra-lightweight construction. These fast-steering mirrors are used to continuously correct for atmospheric disturbances and telescope vibrations. An example of this is the European Extremely Large Telescope (E-ELT) M5 lightweight mirror, which is part of the Tip-Tilt/Field-Stabilization Unit. The baseline design for the E-ELT M5 mirror, as presented in the E-ELT Construction Proposal, is a closed-back ULE mirror with a lightweight core using square core cells. Corning Incorporated (Corning) has a long history of manufacturing lightweight mirror blanks using ULE in a closed-back construction, going back to the 1960's, and includes the Hubble Space Telescope primary mirror, Subaru Telescope secondary and tertiary mirrors, the Magellan I and II tertiary mirrors, and Kepler Space Telescope primary mirror, among many others. A parametric study of 1-meter class lightweight mirror designs showed that Corning's capability to seal a continuous back sheet to a light-weighted core structure provides superior mirror rigidity, in a near-zero thermal expansion material, relative to other existing technologies in this design space. Corning has investigated the parametric performance of several design characteristics for a 3-meter class lightweight mirror blank for the E-ELT M5. Finite Element Analysis was performed on several design scenarios to obtain weight, areal density, and first Eigen frequency. This paper presents an overview of Corning ULE and lightweight mirror manufacturing capabilities, the parametric performance of design characteristics for 1-meter class and 3-meter class lightweight mirrors, as well as the manufacturing advantages and disadvantages of those characteristics.

  5. Strain-balanced type-II superlattices for efficient multi-junction solar cells.

    Science.gov (United States)

    Gonzalo, A; Utrilla, A D; Reyes, D F; Braza, V; Llorens, J M; Fuertes Marrón, D; Alén, B; Ben, T; González, D; Guzman, A; Hierro, A; Ulloa, J M

    2017-06-21

    Multi-junction solar cells made by assembling semiconductor materials with different bandgap energies have hold the record conversion efficiencies for many years and are currently approaching 50%. Theoretical efficiency limits make use of optimum designs with the right lattice constant-bandgap energy combination, which requires a 1.0-1.15 eV material lattice-matched to GaAs/Ge. Nevertheless, the lack of suitable semiconductor materials is hindering the achievement of the predicted efficiencies, since the only candidates were up to now complex quaternary and quinary alloys with inherent epitaxial growth problems that degrade carrier dynamics. Here we show how the use of strain-balanced GaAsSb/GaAsN superlattices might solve this problem. We demonstrate that the spatial separation of Sb and N atoms avoids the ubiquitous growth problems and improves crystal quality. Moreover, these new structures allow for additional control of the effective bandgap through the period thickness and provide a type-II band alignment with long carrier lifetimes. All this leads to a strong enhancement of the external quantum efficiency under photovoltaic conditions with respect to bulk layers of equivalent thickness. Our results show that GaAsSb/GaAsN superlattices with short periods are the ideal (pseudo)material to be integrated in new GaAs/Ge-based multi-junction solar cells that could approach the theoretical efficiency limit.

  6. Regulatory T cells control strain specific resistance to Experimental Autoimmune Prostatitis.

    Science.gov (United States)

    Breser, Maria L; Lino, Andreia C; Motrich, Ruben D; Godoy, Gloria J; Demengeot, Jocelyne; Rivero, Virginia E

    2016-09-14

    Susceptibility to autoimmune diseases results from the encounter of a complex and long evolved genetic context with a no less complex and changing environment. Major actors in maintaining health are regulatory T cells (Treg) that primarily dampen a large subset of autoreactive lymphocytes escaping thymic negative selection. Here, we directly asked whether Treg participate in defining susceptibility and resistance to Experimental Autoimmune Prostatitis (EAP). We analyzed three common laboratory strains of mice presenting with different susceptibility to autoimmune prostatitis upon immunization with prostate proteins. The NOD, the C57BL/6 and the BALB/c mice that can be classified along a disease score ranging from severe, mild and to undetectable, respectively. Upon mild and transient depletion of Treg at the induction phase of EAP, each model showed an increment along this score, most remarkably with the BALB/c mice switching from a resistant to a susceptible phenotype. We further show that disease associates with the upregulation of CXCR3 expression on effector T cells, a process requiring IFNγ. Together with recent advances on environmental factors affecting Treg, these findings provide a likely cellular and molecular explanation to the recent rise in autoimmune diseases incidence.

  7. Factors invovled in L. paracasei subp. paracasei M5-L inhibition of the Shigella sonnei adhesion%副干酪乳杆菌M5-L抑制宋内志贺氏菌黏附作用影响因素的研究

    Institute of Scientific and Technical Information of China (English)

    张英春; 马微; 易华西; 张兰威

    2012-01-01

    研究副干酪乳杆菌M5-L抑制宋内志贺氏菌(S.sonnei)对HT-29细胞黏附的作用及其影响抑制黏附作用的表面分子分析。结果表明M5-L能够显著的抑制S.sonnei的黏附作用,在排除、竞争和取代黏附实验中,抑制率分别为55%、33%和30%,差异显著(p〈0.05),高碘酸钠处理后,M5-L的抑制黏附作用没有变化,表明碳水化合物并没有参与到黏附抑制过程中。而经过5mol/LLiCl处理后,M5-L的抑制黏附作用下降,说明S-层蛋白参与了M5-L抑制S.sonnei对HT-29细胞的黏附作用。此外,M5-L的S-层蛋白能够显著抑制S.sonnei的黏附作用。%L.paracasei subp. paracasei M5-L inhibition of Shigella sonnei adhesion to HT-29 cells and the type of molecules in the M5-L cells surface which effecting the inhibition of S.sonnei were investigated. In exclusion, competition and displacement assays, L.paracasei subp. paracasei M5-L exhibited significant inhibitory activity (p〈0.05),preventing 55% ,33% and 30% of the S.sonnei cells from adhering to HT-29 cell respectively. Treatment with metaperiodate on the M5-L did not affect the inhibitory activity,which showed that the carbohydrates did not involved in the inhibition adherence. When the M5-L treated with 5mol/L LiCI,the inhibition activity was decreased significantly,which indicated the S-layer protein was the important molecules in the inhibition activity. In addition,the S-layer proteins exhibited strongly inhibitory effects on the S.sonnei adherence to HT-29 ceils.

  8. Phase evolution and microwave dielectric properties of A5M5O17-type ceramics

    Directory of Open Access Journals (Sweden)

    Ali Murad

    2017-07-01

    Full Text Available A number of A5M5O17 (A = Na, Ca, Sr, La, Nd, Sm, Gd, Dy, Yb; B = Ti, Nb, Ta type compounds were prepared by a solid-state sintering route and characterized in terms of structure, microstructure and microwave dielectric properties. The compatibility of rare earths with mixed niobate/tantalate and titanate phases was investigated. The larger ionic radii mismatch resulted in the formation of pyrochlore and/or mixed phases while in other cases, pure A5M5O17 phase was formed. The samples exhibited relative permittivity in the range of 35 to 82, quality factor (Q × fo = 897 GHz to 11946 GHz and temperature coefficient of resonance frequency (τf = -120 ppm/°C to 318 ppm/°C.

  9. M5-brane in three-form flux and multiple M2-branes

    CERN Document Server

    Ho, Pei-Ming; Matsuo, Yutaka; Shiba, Shotaro

    2008-01-01

    We investigate the Bagger-Lambert-Gustavsson model associated with the Nambu-Poisson algebra as a theory describing a single M5-brane. We argue that the model is a gauge theory associated with the volume-preserving diffeomorphism in the three-dimenisonal internal space. We derive gauge transformations, actions, supersymmetry transformations, and equations of motions in terms of six-dimensional fields. The equations of motions are written in gauge-covariant form, and the equations for tensor fields have manifest self-dual structure. We demonstrate that the double dimensional reduction of the model reproduces the non-commutative U(1) gauge theory on a D4-brane with a small non-commutativity parameter. We establish relations between parameters in the BLG model and those in M-theory. This shows that the model describes an M5-brane in a large C-field background.

  10. 3d $\\mathcal{N}=2$ minimal SCFTs from Wrapped M5-branes

    CERN Document Server

    Bae, Jin-Beom; Lee, Jaehoon

    2016-01-01

    We study CFT data of 3-dimensional superconformal field theories (SCFTs) arising from wrapped two M5-branes on closed hyperbolic 3-manifolds. Via so-called 3d/3d correspondence, central charges of these SCFTs are related to a $SL(2)$ Chern-Simons (CS) invariant on the 3-manifolds. We give a rigorous definition of the invariant in terms of resurgence theory and a state-integral model for the complex CS theory. We numerically evaluate the central charges for several closed 3-manifolds with small hyperbolic volume. The computation suggests that the wrapped M5-brane systems give infinitely many discrete SCFTs with small central charges. We also analyze these `minimal' SCFTs in the eye of 3d $\\mathcal{N}=2$ superconformal bootstrap.

  11. Riding the Populist Web: Contextualizing the Five Star Movement (M5S in Italy

    Directory of Open Access Journals (Sweden)

    Liza Lanzone

    2015-08-01

    Full Text Available This article focuses on three mechanisms to explain the rise of populist movements across Europe. They are politicization of resentment, exploitation of social cleavages, and polarization of resentment and feelings of non-representation. We conceptualize populism as a strategic power game aiming to transform potential majorities into real ones by creating or reframing social cleavages. Our theoretical model is used to explain the rise of the Five Star Movement (M5S. Beppe Grillo’s M5S gained notoriety on the national political scene in Italy just before the 2013 elections and succeeded in get-ting nearly 25 percent of the overall vote. Moreover, it was the only political force that was able to attract votes across the different regions in Italy, making it the country’s only truly national party.

  12. 奥林巴斯OM—DE—M5数码相机测试

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    奥林巴斯公司在相机制造方面资历资深,沿袭胶片时代”PEN”系列的理念成功推出数码版PEN系列产品后,又以久负盛名的”OM”机型为蓝本.诞生了数码时代的“OM”相机,这就是于2012年2月发布的微型可换镜头相机OM—DE—M5。这个名字有点长.不难看出“OM—D”有数码版OM相机的含义.此外E—M5属于微型4/3系统机型。

  13. Quantifying Susceptibility of CD4+ Stem Memory T-Cells to Infection by Laboratory Adapted and Clinical HIV-1 Strains

    Directory of Open Access Journals (Sweden)

    Jacqueline K. Flynn

    2014-02-01

    Full Text Available CD4+ T cells are principal targets for human immunodeficiency virus type 1 (HIV-1 infection. CD4+ T cell subsets are heterogeneous cell populations, divided by functional and phenotypic differences into naïve and memory T cells. The memory CD4+ T cells are further segregated into central, effector and transitional memory cell subsets by functional, phenotypic and homeostatic characteristics. Defining the distribution of HIV-1 infection in different T cell subsets is important, as this can play a role in determining the size and composition of the viral reservoir. Both central memory and transitional memory CD4+ T cells have been described as long-lived viral reservoirs for HIV. Recently, the newly described stem memory T cell subset has also been implicated as a long-lived HIV reservoir. Using green fluorescent protein (GFP reporter strains of HIV-1 and multi parameter flow cytometry, we developed an assay to simultaneously quantify the susceptibility of stem memory (TSCM, central memory, effector memory, transitional memory and naïve CD4+ T cell subsets, to HIV-1 infection in vitro. We show that TSCM are susceptible to infection with laboratory adapted and clinical HIV-1 strains. Our system facilitates the quantitation of HIV-1 infection in alternative T cell subsets by CCR5- and CXCR4-using viruses across different HIV-1 subtypes, and will be useful for studies of HIV-1 pathogenesis and viral reservoirs.

  14. Quantifying susceptibility of CD4+ stem memory T-cells to infection by laboratory adapted and clinical HIV-1 strains.

    Science.gov (United States)

    Flynn, Jacqueline K; Paukovics, Geza; Cashin, Kieran; Borm, Katharina; Ellett, Anne; Roche, Michael; Jakobsen, Martin R; Churchill, Melissa J; Gorry, Paul R

    2014-02-10

    CD4+ T cells are principal targets for human immunodeficiency virus type 1 (HIV-1) infection. CD4+ T cell subsets are heterogeneous cell populations, divided by functional and phenotypic differences into naïve and memory T cells. The memory CD4+ T cells are further segregated into central, effector and transitional memory cell subsets by functional, phenotypic and homeostatic characteristics. Defining the distribution of HIV-1 infection in different T cell subsets is important, as this can play a role in determining the size and composition of the viral reservoir. Both central memory and transitional memory CD4+ T cells have been described as long-lived viral reservoirs for HIV. Recently, the newly described stem memory T cell subset has also been implicated as a long-lived HIV reservoir. Using green fluorescent protein (GFP) reporter strains of HIV-1 and multi parameter flow cytometry, we developed an assay to simultaneously quantify the susceptibility of stem memory (TSCM), central memory, effector memory, transitional memory and naïve CD4+ T cell subsets, to HIV-1 infection in vitro. We show that TSCM are susceptible to infection with laboratory adapted and clinical HIV-1 strains. Our system facilitates the quantitation of HIV-1 infection in alternative T cell subsets by CCR5- and CXCR4-using viruses across different HIV-1 subtypes, and will be useful for studies of HIV-1 pathogenesis and viral reservoirs.

  15. Functional dissection of Streptococcus pyogenes M5 protein: the hypervariable region is essential for virulence.

    Directory of Open Access Journals (Sweden)

    Johan Waldemarsson

    Full Text Available The surface-localized M protein of Streptococcus pyogenes is a major virulence factor that inhibits phagocytosis, as determined ex vivo. Because little is known about the role of M protein in vivo we analyzed the contribution of different M protein regions to virulence, using the fibrinogen (Fg-binding M5 protein and a mouse model of acute invasive infection. This model was suitable, because M5 is required for mouse virulence and binds mouse and human Fg equally well, as shown here. Mixed infection experiments with wild type bacteria demonstrated that mutants lacking the N-terminal hypervariable region (HVR or the Fg-binding B-repeat region were strongly attenuated, while a mutant lacking the conserved C-repeats was only slightly attenuated. Because the HVR of M5 is not required for phagocytosis resistance, our data imply that this HVR plays a major but unknown role during acute infection. The B-repeat region is required for phagocytosis resistance and specifically binds Fg, suggesting that it promotes virulence by binding Fg. However, B-repeat mutants were attenuated even in Fg-deficient mice, implying that the B-repeats may have a second function, in addition to Fg-binding. These data demonstrate that two distinct M5 regions, including the HVR, are essential to virulence during the early stages of an infection. In particular, our data provide the first in vivo evidence that the HVR of an M protein plays a major role in virulence, focusing interest on the molecular role of this region.

  16. Holography of Wrapped M5-branes and Chern-Simons theory

    OpenAIRE

    Gang, Dongmin; Kim, Nakwoo; Lee, Sangmin

    2014-01-01

    We study three-dimensional superconformal field theories on wrapped M5-branes. Applying the gauge/gravity duality and the recently proposed 3d–3d relation, we deduce quantitative predictions for the perturbative free energy of a Chern–Simons theory on hyperbolic 3-space. Remarkably, the perturbative expansion is expected to terminate at two-loops in the large N limit. We check the correspondence numerically in a number of examples, and confirm the N3 scaling with precise coefficients.

  17. Cell tropism and molecular epidemiology of Anaplasma platys-like strains in cats.

    Science.gov (United States)

    Zobba, R; Anfossi, A G; Visco, S; Sotgiu, F; Dedola, C; Pinna Parpaglia, M L; Battilani, M; Pittau, M; Alberti, A

    2015-04-01

    Bacterial species of the genus Anaplasma are tick transmitted pathogens that negatively impact on animal productions and generate veterinary and public health concerns. This paper reports the identification, molecular characterization and phylogeny of novel unclassified A. platys-like strains in cats. Interestingly, these novel strains are closely related to conspecific strains recently identified in ruminants, and significantly differ from A. platys. A. platys-like strains in cats, unlike ruminants strains, show tropism for platelets. Results have implications in the diagnostic scenario of animal anaplasmosis and provide background for reconstructing the evolutionary history of species genetically related to A. platys. Copyright © 2015 Elsevier GmbH. All rights reserved.

  18. Variable Responses to Carbon Utilization between Planktonic and Biofilm Cells of a Human Carrier Strain of Salmonella enterica Serovar Typhi.

    Directory of Open Access Journals (Sweden)

    Kalaivani Kalai Chelvam

    Full Text Available Salmonella enterica serovar Typhi (S. Typhi is a foodborne pathogen that causes typhoid fever and infects only humans. The ability of S. Typhi to survive outside the human host remains unclear, particularly in human carrier strains. In this study, we have investigated the catabolic activity of a human carrier S. Typhi strain in both planktonic and biofilm cells using the high-throughput Biolog Phenotype MicroArray, Minimum Biofilm Eradication Concentration (MBEC biofilm inoculator (96-well peg lid and whole genome sequence data. Additional strains of S. Typhi were tested to further validate the variation of catabolism in selected carbon substrates in the different bacterial growth phases. The analyzes of the carbon utilization data indicated that planktonic cells of the carrier strain, S. Typhi CR0044 could utilize a broader range of carbon substrates compared to biofilm cells. Pyruvic acid and succinic acid which are related to energy metabolism were actively catabolised in the planktonic stage compared to biofilm stage. On the other hand, glycerol, L-fucose, L-rhamnose (carbohydrates and D-threonine (amino acid were more actively catabolised by biofilm cells compared to planktonic cells. Notably, dextrin and pectin could induce strong biofilm formation in the human carrier strain of S. Typhi. However, pectin could not induce formation of biofilm in the other S. Typhi strains. Phenome data showed the utilization of certain carbon substrates which was supported by the presence of the catabolism-associated genes in S. Typhi CR0044. In conclusion, the findings showed the differential carbon utilization between planktonic and biofilm cells of a S. Typhi human carrier strain. The differences found in the carbon utilization profiles suggested that S. Typhi uses substrates mainly found in the human biliary mucus glycoprotein, gallbladder, liver and cortex of the kidney of the human host. The observed diversity in the carbon catabolism profiles among

  19. Variable Responses to Carbon Utilization between Planktonic and Biofilm Cells of a Human Carrier Strain of Salmonella enterica Serovar Typhi.

    Science.gov (United States)

    Kalai Chelvam, Kalaivani; Yap, Kien Pong; Chai, Lay Ching; Thong, Kwai Lin

    2015-01-01

    Salmonella enterica serovar Typhi (S. Typhi) is a foodborne pathogen that causes typhoid fever and infects only humans. The ability of S. Typhi to survive outside the human host remains unclear, particularly in human carrier strains. In this study, we have investigated the catabolic activity of a human carrier S. Typhi strain in both planktonic and biofilm cells using the high-throughput Biolog Phenotype MicroArray, Minimum Biofilm Eradication Concentration (MBEC) biofilm inoculator (96-well peg lid) and whole genome sequence data. Additional strains of S. Typhi were tested to further validate the variation of catabolism in selected carbon substrates in the different bacterial growth phases. The analyzes of the carbon utilization data indicated that planktonic cells of the carrier strain, S. Typhi CR0044 could utilize a broader range of carbon substrates compared to biofilm cells. Pyruvic acid and succinic acid which are related to energy metabolism were actively catabolised in the planktonic stage compared to biofilm stage. On the other hand, glycerol, L-fucose, L-rhamnose (carbohydrates) and D-threonine (amino acid) were more actively catabolised by biofilm cells compared to planktonic cells. Notably, dextrin and pectin could induce strong biofilm formation in the human carrier strain of S. Typhi. However, pectin could not induce formation of biofilm in the other S. Typhi strains. Phenome data showed the utilization of certain carbon substrates which was supported by the presence of the catabolism-associated genes in S. Typhi CR0044. In conclusion, the findings showed the differential carbon utilization between planktonic and biofilm cells of a S. Typhi human carrier strain. The differences found in the carbon utilization profiles suggested that S. Typhi uses substrates mainly found in the human biliary mucus glycoprotein, gallbladder, liver and cortex of the kidney of the human host. The observed diversity in the carbon catabolism profiles among different S

  20. Differentiation of different mixed Listeria strains and also acid-injured, heat-injured, and repaired cells of Listeria monocytogenes using fourier transform infrared spectroscopy.

    Science.gov (United States)

    Nyarko, Esmond; Donnelly, Catherine

    2015-03-01

    Fourier transform infrared (FT-IR) spectroscopy was used to differentiate mixed strains of Listeria monocytogenes and mixed strains of L. monocytogenes and Listeria innocua. FT-IR spectroscopy was also applied to investigate the hypothesis that heat-injured and acid-injured cells would return to their original physiological integrity following repair. Thin smears of cells on infrared slides were prepared from cultures for mixed strains of L. monocytogenes, mixed strains of L. monocytogenes and L. innocua, and each individual strain. Heat-injured and acid-injured cells were prepared by exposing harvested cells of L. monocytogenes strain R2-764 to a temperature of 56 ± 0.2°C for 10 min or lactic acid at pH 3 for 60 min, respectively. Cellular repair involved incubating aliquots of acid-injured and heat-injured cells separately in Trypticase soy broth supplemented with 0.6% yeast extract for 22 to 24 h; bacterial thin smears on infrared slides were prepared for each treatment. Spectral collection was done using 250 scans at a resolution of 4 cm(-1) in the mid-infrared wavelength region. Application of multivariate discriminant analysis to the wavelength region from 1,800 to 900 cm(-1) separated the individual L. monocytogenes strains. Mixed strains of L. monocytogenes and L. monocytogenes cocultured with L. innocua were successfully differentiated from the individual strains when the discriminant analysis was applied. Different mixed strains of L. monocytogenes were also successfully separated when the discriminant analysis was applied. A data set for injury and repair analysis resulted in the separation of acid-injured, heat-injured, and intact cells; repaired cells clustered closer to intact cells when the discriminant analysis (1,800 to 600 cm(-1)) was applied. FT-IR spectroscopy can be used for the rapid source tracking of L. monocytogenes strains because it can differentiate between different mixed strains and individual strains of the pathogen.

  1. Infections caused by Moraxella, Moraxella urethralis, Moraxella-like groups M-5 and M-6, and Kingella kingae in the United States, 1953-1980.

    Science.gov (United States)

    Graham, D R; Band, J D; Thornsberry, C; Hollis, D G; Weaver, R E

    1990-01-01

    From 1953 to 1980 the Centers for Disease Control received 933 isolates of bacteria belonging to species of the genus Moraxella, Moraxella-like Moraxella urethralis, now renamed Oligella urethralis, unnamed groups M-5 and M-6, and Kingella kingae. Ordinarily sterile sites were the source of 233 isolates. Moraxella nonliquefaciens, the most common isolate (356 strains), was recovered from upper respiratory or ocular sites in 208 (58%) of the cases. Moraxella osloensis was next most common (199 strains) but was the most frequent blood isolate (44 cases). K. kingae appeared especially invasive, with 58 of 78 isolates from blood, bone, or joint. Of the K. kingae strains, 75% were recovered from children under 6 years, compared with 23% of the other strains from that age group (P less than .01). Of the 74 isolates of group M-5, 53 were from wounds caused by dog bites; no other organism in this series was recovered from such wounds. Sixteen of the 28 M. urethralis isolates were from urine. Cases occurred as single infections, with no evidence of clusters. Of patients with infection of ordinarily sterile sites, 9.3% died; only bacteremia, meningitis, and empyema caused fatalities. Most referring laboratories (98%) had not identified the organisms to species, and only 30% had identified them to correct genus. Susceptibility testing by broth dilution revealed low MICs of penicillin (mean, 0.3; 64% less than 1 micrograms/mL). Moraxella, M. urethralis, M-5, M-6, and Kingella are important but frequently misidentified pathogens for humans; penicillin appears to be the treatment of choice.

  2. [Extracellular hydrolases of strain Bacillus sp. 739 and their involvement in the lysis of micromycete cell walls].

    Science.gov (United States)

    Aktuganov, G E; Galimzianova, N F; Melent'ev, A I; Kuz'mina, L Iu

    2007-01-01

    The mycolytic bacterial strain Bacillus sp. 739 produces extracellular enzymes which degrade in vitro the cell walls of a number of phytopathogenic and saprophytic fungi. When Bacillus sp. 739 was cultivated with Bipolaris sorokiniana, a cereal root-rot pathogen, the fungus degradation process correlated with the levels of the beta-1,3-glucanase and protease activity. The comparative characteristic of Bacillus sp. 739 enzymatic preparations showed that efficient hydrolysis of the fungus cell walls was the result of the action of the complex of enzymes produced by the strain when grown on chitin-containing media. Among the enzymes of this complex, chitinases and beta-1,3-glucanases hydrolyzed most actively the disintegrated cell walls of B. sorokiniana. However, only beta-1,3-glucanases were able to degrade the cell walls of native fungal mycelium in the absence of other hydrolases, which is indicative of their key role in the mycolytic activity of Bacillus sp. 739.

  3. High cell density propionic acid fermentation with an acid tolerant strain of Propionibacterium acidipropionici.

    Science.gov (United States)

    Wang, Zhongqiang; Jin, Ying; Yang, Shang-Tian

    2015-03-01

    Propionic acid is an important chemical with wide applications and its production via fermentation is of great interest. However, economic production of bio-based propionic acid requires high product titer, yield, and productivity in the fermentation. A highly efficient and stable high cell density (HCD) fermentation process with cell recycle by centrifugation was developed for propionic acid production from glucose using an acid-tolerant strain of Propionibacterium acidipropionici, which had a higher specific growth rate, productivity, and acid tolerance compared to the wild type ATCC 4875. The sequential batch HCD fermentation at pH 6.5 produced propionic acid at a high titer of ∼40 g/L and productivity of 2.98 g/L h, with a yield of ∼0.44 g/g. The product yield increased to 0.53-0.62 g/g at a lower pH of 5.0-5.5, which, however, decreased the productivity to 1.28 g/L h. A higher final propionic acid titer of >55 g/L with a productivity of 2.23 g/L h was obtained in fed-batch HCD fermentation at pH 6.5. A 3-stage simulated fed-batch process in serum bottles produced 49.2 g/L propionic acid with a yield of 0.53 g/g and productivity of 0.66 g/L h. These productivities, yields and propionic acid titers were among the highest ever obtained in free-cell propionic acid fermentation.

  4. Strain-dependent differences in sensitivity of rat beta-cells to interleukin 1 beta in vitro and in vivo

    DEFF Research Database (Denmark)

    Reimers, J I; Andersen, H U; Mauricio, D

    1996-01-01

    The aim of this study was to investigate whether strain-dependent differences in beta-cell sensitivity to interleukin (IL) 1 beta exist in vitro and in vivo and if so, whether these differences correlate to variations in IL-1 beta-induced islet inducible nitric oxide synthase (iNOS) mRNA expressi...

  5. 羊布鲁杆菌M5-90 BP26抗原单克隆抗体的制备及鉴定%Preparation and characterization of monoclonal antibodies against BP26 protein of Brucella melitensis M5-90

    Institute of Scientific and Technical Information of China (English)

    丘金浪; 吴静波; 黎诚耀; 王文敬

    2012-01-01

    Objective To prepare high specific monoclonal antibodies(mAbs) against BP26 of Brucella(B.)melitensis.Methods A recombinant plasmid pET-28a-BP26 was constructed and transformed into competent Escherichia coli BL21 (DE3),and then the bacteria were induced by 1 mmol/L isopropylthio-β-D-galactoside (IPTG).After induction,the recombinant BP26 protein (rBP26) was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PGAE) and nickel ion affinity chromatography(Ni-NTA).Mice were inoculated with rBP26 antigens for three times at 2-week intervals.The first subcutaneous injection contained 100 μg rBP26 with 0.1 ml complete Freund adjuvant.The second subcutaneous injection was 50 μg rBP26 with 0.1 ml incomplete Freund adjuvant.The antibody titers to rBP26 were determined 2 weeks after each reimmunization.Three days before cell fusion,the mice with the highest titer were intraperitoneally injected with 50 μg rBP26 in 0.1 ml PBS.Pre- and post-immunization sera were collected and used as negative or positive controls for screening mAbs.Mice with the highest titer were sacrificed and spleen cells were isolated.The spleen cells of rBP26 immunized mice were fused with SP2/0 myeloma cells in a ratio of 5 ∶ 1 by polyethylene glycol(PEG) 1450.Antibody-producing hybridomas were primarily screened by an indirect enzyme-linked immunosorbnent assay(ELISA) with rBP26.Reactive hybridomas were subcloned for 3 times,then the strains of hybridoma cells secreting antibodies against BP26 were obtained.Supernatant of cloned hybridoma cultures was collected for mAb analyses.These mAbs were named by the hybridoma clone number and tested their reactivity to membrane proteins extracted(NMP) from B.melitensis vaccine strain(M5-90) by Western blotting and Dot-ELISA.mAbs isotyping and kappa(κ) or lambda(λ) light chain was identified by Mouse Monoclonal Antibody Isotyping Kit.Results A total of two mAbs reactive to rBP26 of B.melitensis were selected from antibody screening

  6. San Andreas Fault, California, M 5.5 or greater Earthquakes 1800-2000

    Science.gov (United States)

    Toppozada, T.; Branum, D.; Reichle, M.; Hallstrom, C.

    2001-12-01

    The San Andreas fault has been the most significant source of major California earthquakes since 1800. From 1812 to 1906 it generated four major earthquakes of M 7.2 or greater in two pairs on two major regions of the fault. A pair of major earthquakes occurred on the Central to Southern region, where the 1857 faulting overlapped the 1812 earthquake faulting. And a pair of major earthquakes occurred on the Northern region, where the 1906 faulting overlapped the 1838 earthquake faulting. The 1812 earthquake resulted from a rupture of up to about 200 km, from the region of Cajon Pass to as far as about 50 km west of Fort Tejon (Sieh and others, 1989). This rupture is the probable source of both the destructive 1812.12.8 "San Juan Capistrano" and the 1812.12.21 "Santa Barbara Channel" earthquakes. The 1838 earthquake's damage effects throughout the Bay area, from San Francisco to Santa Clara Valley and Monterey, were unequalled by any Bay area earthquake other than the 1906 event. The mainshock's effects, and numerous strong probable aftershocks in the San Juan Bautista vicinity in the following three years, suggest 1838 faulting from San Francisco to San Juan Bautista, and M about 7.4. The 630 km length of the San Andreas fault between San Francisco and Cajon Pass ruptured in the 1838 and 1857 earthquakes, except for about 75 km between Bitterwater and San Juan Bautista. The 1840-1841 probable aftershocks of the 1838 event occurred near San Juan Bautista, and the foreshocks and aftershocks of the 1857 event occurred near Bitterwater. In the Bitterwater area, strong earthquakes continued to occur until the 1885 earthquake of M 6.5. Near Parkfield, 40 to 70 km southeast of Bitterwater, M 5.5 or greater earthquakes have occurred from the 1870s to the 1960s. In the total Bitterwater to Parkfield zone bracketing the northern end of the 1857 rupture, the seismicity and moment release has decreased steadily since 1857, and has tended to migrate southeastward with time. The

  7. The potential for archiving and reconstituting valuable strains of turkey (Meleagris gallopavo) using primordial germ cells.

    Science.gov (United States)

    Wade, Alexander J; French, Nick A; Ireland, Grenham W

    2014-04-01

    Diseases such as avian influenza can destroy turkey flocks, potentially resulting in the loss of valuable or rare genetic material. Consequently, there is an urgent need to develop a means to archive such germplasm. Germline chimeras produced by intravascular transfer of primordial germ cells (PGC) have been reported in other avian species but not turkeys. This study examined the feasibility of both establishing an archive of frozen PGC, and producing germline chimeras by injecting the thawed PGC into host embryos. To meet these aims, the following experiments were performed: (1) PGC identification within turkey embryos; (2) development of an efficient method for isolation of turkey PGC; (3) demonstration that PGC can be cryopreserved, recovered, and retain viability; (4) reinjection into embryos and detection of injected PGC. Primordial germ cells were identified using periodic acid-Schiff reagent and the immunological marker OLP-1. Bloodstream PGC were isolated using Ficoll density gradient centrifugation with PGC recovery peaking at stages 13, 14, and 15 with 32 ± 4.9, 33 ± 6.4, and 26 ± 5.4 PGC recovered, respectively. Primordial germ cells were frozen using Dulbecco's modified Eagle medium, 20% fetal calf serum, and 10% dimethylsulfoxide and demonstrated 90 ± 1.7% viability after 3 mo frozen in liquid nitrogen. Freshly isolated and frozen thawed DiI- and Q-Tracker-labeled PGC repopulated stage 30 gonads after vascular transfer into ex ovo cultured embryos. The DiI-labeled cells repopulated gonads less frequently, with 36 ± 13.2% of gonads containing the DiI-labeled PGC, and 7 ± 3.8% of reinjected PGC reaching the gonads of positive embryos. The Q-tracker-labeled cells were detected more frequently in embryos, with 67 ± 21.1% having positive signals, and 44 ± 4.9% of reinjected Q-tracker-labeled PGC colonized the gonads of positive embryos. This study demonstrated the feasibility of using turkey PGC to archive turkey germplasm from different strains

  8. Effect of different feed ingredients and additives on IPEC-J2 cells challenged with an enterotoxigenic Escherichia coli strain.

    Science.gov (United States)

    Spitzer, F; Speiser, S; Vahjen, W; Zentek, J

    2016-08-01

    The intestinal porcine epithelial cell line IPEC-J2 was used as an in vitro model to assess effects of additives on the adhesion and cell toxic effects of a F4-positive (ETEC) and a F4-negative Escherichia coli (DSM 2840) strain. Bacterial adhesion was examined using flow cytometry in IPEC-J2 cells infected with bacteria stained with 5,6-carboxymethyl fluorescein diacetate succinimidyl ester. Measurement of transepithelial electrical resistance (TEER) was performed to characterize the impact on IPEC-J2 monolayer integrity. The feed additives were prepared as aqueous extract and tested in different dilutions and incubation times. The F4-positive ETEC strain had a high adhesion to IPEC-J2 cells and reduced TEER shortly after the in vitro infection. The nonpathogenic E. coli strain DSM 2840 showed only low adhesion capacity and no TEER impairment. Infection with ETEC with added test extracts showed a reduction of bacterial adhesion to IPEC-J2 cells by an autolyzed yeast product (p < 0.05). Bovine colostrum, an additive containing thyme extract and an organic acid mix did not interfere with the ETEC adherence. The TEER decrease of the IPEC-J2 monolayer after ETEC infection was not affected by the added substances. In conclusion, interference with epithelial adhesion might be a protective mechanism of the tested yeast extract, indicating that the cell culture model might be suitable as screening tool to complement in vivo challenge trials with piglets.

  9. Assessment of ventricular function in adults with sickle cell disease: role of two-dimensional speckle-tracking strain.

    Science.gov (United States)

    Barbosa, Marcia M; Vasconcelos, Maria Carmen M; Ferrari, Teresa Cristina A; Fernandes, Bárbara Martins; Passaglia, Luiz Guilherme; Silva, Célia Maria; Nunes, Maria Carmo P

    2014-11-01

    Sickle cell disease (SCD) is a hemoglobinopathy that is common worldwide. It usually presents with cardiac involvement, although data on systolic function are somewhat controversial. The aim of this study was to investigate the value of speckle-tracking strain, a deformation index, in detecting ventricular dysfunction in SCD. Ninety adult patients with SCD were compared with 20 healthy controls. Doppler echocardiography with Doppler tissue imaging was performed in all, and the left and right ventricles were analyzed by the use of two-dimensional speckle-tracking strain. The mean age of the patients with SCD was 26 years, and 43% were men. Left ventricular (LV) dimensions and mass were higher in patients with SCD, whereas LV ejection fraction did not differ from the controls. E and A waves, as well as E/e' ratio, were also higher in patients with SCD. Two-dimensional speckle-tracking strain of both ventricles in the patients with SCD was not different from that of controls. The factors independently associated with LV longitudinal strain were age (P = .009), oximetry (P = .001), lactate dehydrogenase (P = .014), LV ejection fraction (P speckle-tracking strain of both ventricles was similar in patients and controls, suggesting normal myocardial contractility in patients with SCD. LV global longitudinal strain was associated with age, intensity of hemolysis, and ventricular function. Copyright © 2014 American Society of Echocardiography. Published by Elsevier Inc. All rights reserved.

  10. Single strain isolation method for cell culture-adapted hepatitis C virus by end-point dilution and infection.

    Directory of Open Access Journals (Sweden)

    Nao Sugiyama

    Full Text Available The hepatitis C virus (HCV culture system has enabled us to clarify the HCV life cycle and essential host factors for propagation. However, the virus production level of wild-type JFH-1 (JFH-1/wt is limited, and this leads to difficulties in performing experiments that require higher viral concentrations. As the cell culture-adapted JFH-1 has been reported to have robust virus production, some mutations in the viral genome may play a role in the efficiency of virus production. In this study, we obtained cell culture-adapted virus by passage of full-length JFH-1 RNA-transfected Huh-7.5.1 cells. The obtained virus produced 3 log-fold more progeny viruses as compared with JFH-1/wt. Several mutations were identified as being responsible for robust virus production, but, on reverse-genetics analysis, the production levels of JFH-1 with these mutations did not reach the level of cell culture-adapted virus. By using the single strain isolation method by end-point dilution and infection, we isolated two strains with additional mutations, and found that these strains have the ability to produce more progeny viruses. On reverse-genetics analysis, the strains with these additional mutations were able to produce robust progeny viruses at comparable levels as cell culture-adapted JFH-1 virus. The strategy used in this study will be useful for identifying strains with unique characteristics, such as robust virus production, from a diverse population, and for determining the responsible mutations for these characteristics.

  11. Studies on superoxide dismutase activities in virulent and avirulent strains of Agrobacterium tumefaciens and also in normal and crown gall tumor cells of Bryophyllum calycinum.

    Science.gov (United States)

    Banerjee, D; Basu, M; Choudhury, I; Chatterjee, G C

    1982-01-01

    Superoxide dismutase activity in virulent strains of Agrobacterium tumefaciens was found to be higher than that in avirulent strains. Polyacrylamide gel electrophoresis revealed two isoenzymes in both these strains. These isoenzymes are suggested to be iron and manganese containing superoxide dismutases. Crown gall tumor cells of the plant Bryophyllum calycinum were found to have higher superoxide dismutase activity than the normal plant cells. Polyacrylamide gel electrophoresis revealed two isoenzymes in both normal and crown gall tumor cells. Advantages of the higher superoxide dismutase activities in respect of the survival of virulent strains of A. tumefaciens and crown gall tumor growth have been discussed.

  12. Periodontal Ligament Stem Cells in the Periodontitis Microenvironment Are Sensitive to Static Mechanical Strain

    Directory of Open Access Journals (Sweden)

    Jia Liu

    2017-01-01

    Full Text Available During orthodontic treatment, periodontium remodeling of periodontitis patients under mechanical force was abnormal. We have previously confirmed the function impairment of periodontal ligament stem cells (PDLSCs in the periodontitis microenvironment which might be involved in this pathological process. However, the response of PDLSCs in periodontitis microenvironment to mechanical force remains unclear. Therefore, in the present study, we introduced a Flexcell tension apparatus and investigated the response of PDLSCs obtained from periodontal tissues of periodontitis patients (PPDLSCs and of those obtained from healthy periodontal tissues (HPDLSCs to different magnitudes of static mechanical strain (SMS. PPDLSCs showed increased proliferation, decreased osteogenic activity, activated osteoclastogenesis, and greater secretion of inflammatory cytokines. Different magnitudes of SMS exerted distinct effects on HPDLSCs and PPDLSCs. An SMS of 12% induced optimal effects in HPDLSCs, including the highest proliferation, the best osteogenic ability, the lowest osteoclastogenesis, and the lowest secretion of inflammatory cytokines, while the optimal SMS for PPDLSCs was 8%. Excessive SMS damaged PPDLSCs function, including decreased proliferation, an imbalance between osteogenesis and osteoclastogenesis, and an activated inflammatory response. Our data suggest that PPDLSCs are more sensitive and less tolerant to SMS, and this may explain why mechanical force results in undesirable effects in periodontitis patients.

  13. Periodontal Ligament Stem Cells in the Periodontitis Microenvironment Are Sensitive to Static Mechanical Strain

    Science.gov (United States)

    Liu, Jia; Liu, Shiyu; Gao, Jie; Qin, Wen; Song, Yang

    2017-01-01

    During orthodontic treatment, periodontium remodeling of periodontitis patients under mechanical force was abnormal. We have previously confirmed the function impairment of periodontal ligament stem cells (PDLSCs) in the periodontitis microenvironment which might be involved in this pathological process. However, the response of PDLSCs in periodontitis microenvironment to mechanical force remains unclear. Therefore, in the present study, we introduced a Flexcell tension apparatus and investigated the response of PDLSCs obtained from periodontal tissues of periodontitis patients (PPDLSCs) and of those obtained from healthy periodontal tissues (HPDLSCs) to different magnitudes of static mechanical strain (SMS). PPDLSCs showed increased proliferation, decreased osteogenic activity, activated osteoclastogenesis, and greater secretion of inflammatory cytokines. Different magnitudes of SMS exerted distinct effects on HPDLSCs and PPDLSCs. An SMS of 12% induced optimal effects in HPDLSCs, including the highest proliferation, the best osteogenic ability, the lowest osteoclastogenesis, and the lowest secretion of inflammatory cytokines, while the optimal SMS for PPDLSCs was 8%. Excessive SMS damaged PPDLSCs function, including decreased proliferation, an imbalance between osteogenesis and osteoclastogenesis, and an activated inflammatory response. Our data suggest that PPDLSCs are more sensitive and less tolerant to SMS, and this may explain why mechanical force results in undesirable effects in periodontitis patients. PMID:28316629

  14. Zircaloy-4 and M5 high temperature oxidation and nitriding in air

    Energy Technology Data Exchange (ETDEWEB)

    Duriez, C. [Institut de Radioprotection et Surete Nucleaire, Direction de Prevention des Accidents Majeurs, Centre de Cadarache, 13115 St Paul Lez Durance (France)], E-mail: christian.duriez@irsn.fr; Dupont, T.; Schmet, B.; Enoch, F. [Universite Technologique de Troyes, BP 2060, 10010 Troyes (France)

    2008-10-15

    For the purpose of nuclear power plant severe accident analysis, degradation of Zircaloy-4 and M5 cladding tubes in air at high temperature was investigated by thermo-gravimetric analysis, in isothermal conditions, in a 600-1200 deg. C temperature range. Alloys were investigated either in a 'as received' bare state, or after steam pre-oxidation at 500 {sup o}C to simulate in-reactor corrosion. At the beginning of air exposure, the oxidation rate obeys a parabolic law, characteristic of solid-state diffusion limited regime. Parabolic rate constants compare, for Zircaloy-4 as well as for M5, with recently assessed correlations for high temperature Zircaloy-4 steam-oxidation. A thick layer of dense protective zirconia having a columnar structure forms during this diffusion-limited regime. Then, a kinetic transition (breakaway type) occurs, due to radial cracking along the columnar grain boundaries of this protective dense oxide scale. The breakaway is observed for a scale thickness that strongly increases with temperature. At the lowest temperatures, the M5 alloy appears to be breakaway-resistant, showing a delayed transition compared to Zircaloy-4. However, for both alloys, a pre-existing corrosion scale favours the transition, which occurs much earlier. The post transition kinetic regime is linear only for the lowest temperatures investigated. From 800 deg. C, a continuously accelerated regime is observed and is associated with formation of a strongly porous non-protective oxide. A mechanism of nitrogen-assisted oxide growth, involving formation and re-oxidation of ZrN particles, as well as nitrogen associated zirconia phase transformations, is proposed to be responsible for this accelerated degradation.

  15. Status of E-ELT M5 scale-one demonstrator

    Science.gov (United States)

    Barriga, Pablo; Sedghi, Babak; Dimmler, Martin; Kornweibel, Nick

    2014-07-01

    The fifth mirror of the European Extremely Large Telescope optical train is a field stabilization tip/tilt unit responsible for correcting the dynamical tip and tilt caused mainly by wind load on the telescope. A scale-one prototype including the inclined support, the fixed frame and a basic control system was designed and manufactured by NTE-SENER (Spain) and CSEM (Switzerland) as part of the prototyping and design activities. All interfaces to the mirror have been reproduced on a dummy structure reproducing the inertial characteristics of the optical element. The M5 unit is required to have sufficient bandwidth for tip/tilt reference commands coming from the wavefront control system. Such a bandwidth can be achieved using local active damping loop to damp the low frequency mechanical modes before closing a position loop. Prototyping on the M5 unit has been undertaken in order to demonstrate the E-ELT control system architecture, concepts and development standards and to further study active damping strategies. The control system consists of two nested loops: a local damping loop and a position loop. The development of this control system was undertaken following the E-ELT control system development standards in order to determine their applicability and performance and includes hardware selection, communication, synchronization, configuration, and data logging. In this paper we present the current status of the prototype M5 control system and the latest results on the active damping control strategy, in particular the promising results obtained with the method of positive position feedback.

  16. Interactions between rye (Secale cereale) root border cells (RBCs) and pathogenic and nonpathogenic rhizosphere strains of Fusarium culmorum.

    Science.gov (United States)

    Jaroszuk-Sciseł, Jolanta; Kurek, Ewa; Rodzik, Beata; Winiarczyk, Krystyna

    2009-10-01

    Interactions of rye (Secale cereale) root border cells (RBCs), generated during plant growth and surrounding the root cap, with nonpathogenic rhizosphere Fusarium culmorum isolates: DEMFc2 (PGPF) and DEMFc5 (DRMO), and a pathogenic strain DEMFc37 were studied in test tube experiments. The effect of water-suspended RBCs released from the rye root cap on the rate of macroconidia germination and hyphae (mycelial) growth of F. culmorum strains was also examined. It was found that root caps of 3-d-old rye seedlings (with the root length of 20mm) were surrounded with a layer of RBCs generated in a number specific for this plant species of 1980+/-30. Introduction of the macroconidia of the tested F. culmorum strains into the root zone of 3-d-old seedlings resulted, after 3d of incubation, in the formation of mantle-like structures only in the rhizosphere of plants inoculated with the pathogenic DEMFc37 strain. The macroconidia were suspended in (1) water, (2) a water mixture with root caps deprived of RBCs, (3) Martin medium, (4) malt extract broth, and (5) a water mixture with rye RBCs, and their percentage germination was determined during 96-h incubation at 20 degrees C. Germination of the macroconidia of all the tested F. culmorum strains suspended in the rich growth media (Martin and malt extract broth) and in the mixture with RBCs was significantly speeded up. While only an average of 16.6 % of macroconidia suspended in water germinated after 96-h incubation, more than 90 % of those suspended in the growth media or in the mixture with RBCs germinated after 24h of incubation. In all the treatments, the highest rate of macroconidia germination was found in suspensions of the pathogenic strain and the lowest in macroconidial suspensions of the PGPF strain. The stimulatory effect of RBCs was not specific to the pathogenic strain. Nevertheless, microscopic observation revealed that it was only in the suspension containing a mixture of rye RBCs and macroconidia of the

  17. Enhanced Replication of Hepatitis E Virus Strain 47832c in an A549-Derived Subclonal Cell Line

    Science.gov (United States)

    Schemmerer, Mathias; Apelt, Silke; Trojnar, Eva; Ulrich, Rainer G.; Wenzel, Jürgen J.; Johne, Reimar

    2016-01-01

    Hepatitis E virus (HEV) is a human pathogen with increasing importance. The lack of efficient cell culture systems hampers systematic studies on its replication cycle, virus neutralization and inactivation. Here, several cell lines were inoculated with the HEV genotype 3c strain 47832c, previously isolated from a chronically infected transplant patient. At 14 days after inoculation the highest HEV genome copy numbers were found in A549 cells, followed by PLC/PRF/5 cells, whereas HepG2/C3A, Huh-7 Lunet BLR and MRC-5 cells only weakly supported virus replication. Inoculation of A549-derived subclone cell lines resulted in most cases in reduced HEV replication. However, the subclone A549/D3 was susceptible to lower virus concentrations and resulted in higher virus yields as compared to parental A549 cells. Transcriptome analysis indicated a downregulation of genes for carcinoembryonic antigen-related cell adhesion molecules (CEACAM) 5 and 6, and an upregulation of the syndecan 2 (SDC2) gene in A549/D3 cells compared to A549 cells. However, treatment of A549/D3 cells or A549 cells with CEACAM- or syndecan 2-specific antisera did not influence HEV replication. The results show that cells supporting more efficient HEV replication can be selected from the A549 cell line. The specific mechanisms responsible for the enhanced replication remain unknown. PMID:27690085

  18. Holography of wrapped M5-branes and Chern–Simons theory

    Energy Technology Data Exchange (ETDEWEB)

    Gang, Dongmin [School of Physics, Korea Institute for Advanced Study, Seoul 130-722 (Korea, Republic of); Kim, Nakwoo [Department of Physics, Research Institute of Basic Science, Kyung Hee University, Seoul 130-701 (Korea, Republic of); Lee, Sangmin [Center for Theoretical Physics, College of Liberal Studies, Seoul National University, Seoul 151-742 (Korea, Republic of); School of Physics, Korea Institute for Advanced Study, Seoul 130-722 (Korea, Republic of)

    2014-06-02

    We study three-dimensional superconformal field theories on wrapped M5-branes. Applying the gauge/gravity duality and the recently proposed 3d–3d relation, we deduce quantitative predictions for the perturbative free energy of a Chern–Simons theory on hyperbolic 3-space. Remarkably, the perturbative expansion is expected to terminate at two-loops in the large N limit. We check the correspondence numerically in a number of examples, and confirm the N{sup 3} scaling with precise coefficients.

  19. New variables in M5 (NGC 5904) and some identification corrections

    CERN Document Server

    Ferro, A Arellano; Giridhar, S; Luna, A; Muneer, S

    2015-01-01

    We report twelve variables not previously detected in the globular cluster M5 (NGC 5904); one SX Phe and eleven semi-regular variables (SR). Their identifications, equatorial coordinates, ephemerides, and light curves are given. Furthermore, we have explored the light curves of a group of stars whose variability has not been confirmed and that are marked as probable non- variables in the CVSGC. Finally, we offer detailed identifications for some of the known variables in crowded regions that were misidentified in previous studies. We shall also address the cases of the cataclysmic variable or U Gem type V101 and of the variable blue straggler V159.

  20. Full elastic strain and stress tensor measurements from individual dislocation cells in copper through-Si vias.

    Science.gov (United States)

    Levine, Lyle E; Okoro, Chukwudi; Xu, Ruqing

    2015-11-01

    Nondestructive measurements of the full elastic strain and stress tensors from individual dislocation cells distributed along the full extent of a 50 µm-long polycrystalline copper via in Si is reported. Determining all of the components of these tensors from sub-micrometre regions within deformed metals presents considerable challenges. The primary issues are ensuring that different diffraction peaks originate from the same sample volume and that accurate determination is made of the peak positions from plastically deformed samples. For these measurements, three widely separated reflections were examined from selected, individual grains along the via. The lattice spacings and peak positions were measured for multiple dislocation cell interiors within each grain and the cell-interior peaks were sorted out using the measured included angles. A comprehensive uncertainty analysis using a Monte Carlo uncertainty algorithm provided uncertainties for the elastic strain tensor and stress tensor components.

  1. Global gene expression profiling reveals similarities and differences among mouse pluripotent stem cells of different origins and strains

    Science.gov (United States)

    Sharova, Lioudmila V.; Sharov, Alexei A.; Piao, Yulan; Shaik, Nabeebi; Sullivan, Terry; Stewart, Colin L.; Hogan, Brigid L.M.; Ko, Minoru S.H.

    2007-01-01

    Pluripotent stem cell lines with similar phenotypes can be derived from both blastocysts (embryonic stem cells, ESC) and primordial germ cells (embryonic germ cells, EGC). Here, we present a compendium DNA microarray analysis of multiple mouse ESCs and EGCs from different genetic backgrounds (strains 129 and C57BL/6) cultured under standard conditions and in differentiation-promoting conditions by the withdrawal of Leukemia Inhibitory Factor (LIF) or treatment with retinoic acid (RA). All pluripotent cell lines showed similar gene expression patterns, which separated them clearly from other tissue stem cells with lower developmental potency. Differences between pluripotent lines derived from different sources (ESC vs. EGC) were smaller than differences between lines derived from different mouse strains (129 vs. C57BL/6). Even in the differentiation-promoting conditions, these pluripotent cells showed the same general trends of gene expression changes regardless of their origin and genetic background. These data indicate that ESCs and EGCs are indistinguishable based on global gene expression patterns alone. On the other hand, a detailed comparison between a group of ESC lines and a group of EGC lines identified 20 signature genes whose average expression levels were consistently higher in ESC lines, and 84 signature genes whose average expression levels were consistently higher in EGC lines, irrespective of mouse strains. Similar analysis identified 250 signature genes whose average expression levels were consistently higher in a group of 129 cell lines, and 337 signature genes whose average expression levels were consistently higher in a group of C57BL/6 cell lines. Although none of the genes was exclusively expressed in either ESCs versus EGCs or 129 versus C57BL/6, in combination these signature genes provide a reliable separation and identification of each cell type. Differentiation-promoting conditions also revealed some minor differences between the cell

  2. Development of stress tolerant Saccharomyces cerevisiae strains by metabolic engineering: New aspects from cell flocculation and zinc supplementation.

    Science.gov (United States)

    Cheng, Cheng; Zhang, Mingming; Xue, Chuang; Bai, Fengwu; Zhao, Xinqing

    2017-02-01

    Budding yeast Saccharomyces cerevisiae is widely studied for the production of biofuels from lignocellulosic biomass. However, economic production is currently challenged by the repression of cell growth and compromised fermentation performance of S. cerevisiae strains in the presence of various environmental stresses, including toxic level of final products, inhibitory compounds released from the pretreatment of cellulosic feedstocks, high temperature, and so on. Therefore, it is important to improve stress tolerance of S. cerevisiae to these stressful conditions to achieve efficient and economic production. In this review, the latest advances on development of stress tolerant S. cerevisiae strains are summarized, with the emphasis on the impact of cell flocculation and zinc addition. It was found that cell flocculation affected ethanol tolerance and acetic acid tolerance of S. cerevisiae, and addition of zinc to a suitable level improved stress tolerance of yeast cells to ethanol, high temperature and acetic acid. Further studies on the underlying mechanisms by which cell flocculation and zinc status affect stress tolerance will not only enrich our knowledge on stress response and tolerance mechanisms of S. cerevisiae, but also provide novel metabolic engineering strategies to develop robust yeast strains for biofuels production. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. [Kinetic observation on the invasion into and proliferation in IEC-6 cells of Toxoplasma gondii RH strain tachyzoites in vitro].

    Science.gov (United States)

    Meng, Xiao-li; Yin, Guo-rong; Liu, Hong-li; Wang, Hai-long

    2009-02-28

    To study the invasion and proliferation in IEC-6 cells of Toxoplasma gondii RH strain tachyzoites in vitro. T. gondii tachyzoites of RH strain were co-cultured with IEC-6 cells in vitro, the process of cell adhesion, invasion and proliferation by tachyzoites was observed consecutively with inverted microscope. At 5 min, 10 min, 20 min, 30 min, 1, 2, 4, 6, 12, 24 and 48 h after co-culture, the tachyzoite invasion to IEC-6 and intracellular proliferation were observed with Giemsa-Wright's staining, respectively. The invasive rate of tachyzoites to IEC-6 was counted. T. gondii tachyzoites invaded the IEC-6 cells 5 min after culture, henceforth the invasive rate increased gradually. The invasive rate was about 55.0% at the first hour after culture with 1-5 tachyzoites in one cell. In the second hour after culture, the rate reached highest with 81.8% and there were many pseudocysts emerging. At the same time, tachyzoites invaded the cell nucleus and proliferated in the nucleus. At the 4th hour after culture, the invasive rate began to decrease (80.8 +/- 9.2)%, the pseudocysts began to break and tachyzoites were released to cluster. The clustering tachyzoites increased significantly at the 6th hour. At the 12th hour the clustering tachyzoites decreased and most tachyzoites were free, the number of complete cells decreased obviously. There were only a few cells and pseudocysts left at the 24th hour, and a great quantity of free tachyzoites existed out of the IEC-6 cells. There were plenty of mobile tachyzoites while none of IEC-6 cells existed after 48 h culture. IEC-6 cell may be the suitable target cell of Toxoplasma gondii tachyzoite. The tachyzoites can invade the IEC-6 cells quickly in vitro and proliferate in the plasma and nucleus with a reproductive cycle of about 6 to 12 hrs.

  4. A New Overview of Secular Period Changes of RR Lyrae stars inM5

    CERN Document Server

    Ferro, A Arellano; Kains, N; Luna, A

    2016-01-01

    Secular period variations, $\\beta=\\dot{P}$, in 76 RR Lyrae stars in the globular cluster M5 are analysed using our most recent CCD $V$ photometry and the historical photometric database available in the literature since 1889. This provides a time baseline of up to 118 years for these variables. The analysis was performed using two independent approaches: first, the classical $O-C$ behaviour of the time of maximum light, and second, via a grid $(P,\\beta)$, where the solution producing the minimum scatter in the phased light curve is chosen. The results of the two methods agree satisfactorily. This allowed a new interpretation of the nature of the period changes in many RR Lyrae stars in M5. It is found that in $96\\%$ of the stars studied no irregular or stochastic variations need to be claimed, but that $66\\%$ of the population shows steady period increases or decreases, and that $34\\%$ of the periods seem to have been stable over the last century. The lack of systematic positive or negative period variations ...

  5. Forecasting Shaharchay River Flow in Lake Urmia Basin using Genetic Programming and M5 Model Tree

    Directory of Open Access Journals (Sweden)

    S. Samadianfard

    2017-01-01

    Full Text Available Introduction: Precise prediction of river flows is the key factor for proper planning and management of water resources. Thus, obtaining the reliable methods for predicting river flows has great importance in water resource engineering. In the recent years, applications of intelligent methods such as artificial neural networks, fuzzy systems and genetic programming in water science and engineering have been grown extensively. These mentioned methods are able to model nonlinear process of river flows without any need to geometric properties. A huge number of studies have been reported in the field of using intelligent methods in water resource engineering. For example, Noorani and Salehi (23 presented a model for predicting runoff in Lighvan basin using adaptive neuro-fuzzy network and compared the performance of it with neural network and fuzzy inference methods in east Azerbaijan, Iran. Nabizadeh et al. (21 used fuzzy inference system and adaptive neuro-fuzzy inference system in order to predict river flow in Lighvan river. Khalili et al. (13 proposed a BL-ARCH method for prediction of flows in Shaharchay River in Urmia. Khu et al. (16 used genetic programming for runoff prediction in Orgeval catchment in France. Firat and Gungor (11 evaluated the fuzzy-neural model for predicting Mendes river flow in Turkey. The goal of present study is comparing the performance of genetic programming and M5 model trees for prediction of Shaharchay river flow in the basin of Lake Urmia and obtaining a comprehensive insight of their abilities. Materials and Methods: Shaharchay river as a main source of providing drinking water of Urmia city and agricultural needs of surrounding lands and finally one of the main input sources of Lake Urmia is quite important in the region. For obtaining the predetermined goals of present study, average monthly flows of Shaharchay River in Band hydrometric station has been gathered from 1951 to 2011. Then, two third of mentioned

  6. M5C2 carbide precipitates in a high-Cr martensitic steel

    Science.gov (United States)

    Shen, Yinzhong; Ji, Bo; Zhou, Xiaoling

    2014-05-01

    The precipitate phases in an advanced 11% Cr martensitic steel, expected to be used at 650 °C, have been investigated to understand the effect of precipitates on the creep-rupture strength of the steel. M23C6 and MX precipitates were dominant phases in this steel. Needle-like precipitates with a typical length of 180 nm and width of 20 nm; and metallic-element compositions of 53-74Fe, 16-26Cr, 3-18Ta, 2-8W, and 2-4Co (at%); were observed mainly within the martensite laths of the normalized-and-tempered steel. The needle-like precipitates have been identified as monoclinic carbide M5C2, which is not known to have been reported previously in high chromium steels, or in heat-resistant steels those have been normalized-and-tempered. This indicates that the formation of M5C2 carbides can occur in heat-resistant steels produced under appropriate tempering conditions, and that this does not require long-term isothermal aging or creep testing, in all cases.

  7. Differentiation of strains of varicella-zoster virus by changes in neutral lipid metabolism in infected cells

    Energy Technology Data Exchange (ETDEWEB)

    Jerkofsky, M.; De Siervo, A.J.

    1986-03-01

    Eleven isolates of varicella-zoster virus were tested for their effects on the incorporation of (/sup 14/C)acetate into lipids in infected human embryonic lung cells. By relative percent, all virus isolates demonstrated a shift from polar lipid synthesis to neutral lipid, especially triglyceride, synthesis. By data expressed as counts per minute per microgram of protein, the VZV strains could be separated into two groups: those strains which depressed lipid synthesis and those strains which did not depress, and may even have stimulated, lipid, especially triglyceride, synthesis. These results may be useful in understanding the development of lipid changes seen in children affected with Reye's syndrome following chickenpox.

  8. Acid resistance and response to pH-induced stress in two Lactobacillus plantarum strains with probiotic potential.

    Science.gov (United States)

    Šeme, H; Gjuračić, K; Kos, B; Fujs, Š; Štempelj, M; Petković, H; Šušković, J; Bogovič Matijašić, B; Kosec, G

    2015-01-01

    Two new Lactobacillus plantarum strains, KR6-DSM 28780 and M5 isolated from sour turnip and traditional dried fresh cheese, respectively, were evaluated for species identity, antibiotic susceptibility, resistance to gastrointestinal conditions and adaptive response to low pH. Resistance mechanisms involved in the adaptation to acid-induced stress in these two strains were investigated by quantitative PCR of the atpA, cfa1, mleS and hisD genes. In addition to absence of antibiotic resistance, the two L. plantarum strains showed excellent survival rates at pH values as low as 2.4. Adaptive response to low pH was clearly observed in both strains; strain KR6 was superior to M5, as demonstrated by its ability to survive during 3 h incubation at pH 2.0 upon adaptation to moderately acidic conditions. In contrast, acid adaptation did not significantly affect the survival rate during simulated passage through the gastrointestinal tract. In both strains, induction of histidine biosynthesis (hisD) was upregulated during the acid adaptation response. In addition, significant upregulation of the cfa1 gene, involved in modulation of membrane fatty acid composition, was observed during the adaptation phase in strain KR6 but not in strain M5. Cells adapted to moderately acidic conditions also showed a significantly increased viability after the lyophilisation procedure, a cross-protection phenomenon providing additional advantage in probiotic application.

  9. Aerobic decolorization and degradation of Acid Orange G (AOG) by suspended growing cells and immobilized cells of a yeast strain Candida tropicalis TL-F1.

    Science.gov (United States)

    Tan, Liang; Li, Hua; Ning, Shuxiang; Hao, Jia

    2014-10-01

    In this study, aerobic decolorization and degradation of azo dye Acid Orange G (AOG) by both suspended growing cells and immobilized cells of a yeast strain Candida tropicalis TL-F1 were studied. The effects of different parameters on decolorization of AOG by both growing suspended and immobilized strain TL-F1 were investigated. Furthermore, a possible decolorization mechanism of AOG was proposed through analyzing metabolic intermediates using UV-vis and high-performance liquid chromatography-mass spectrometry (HPLC-MS) methods. Strain TL-F1 could decolorize AOG in both liquid and solid mediums through degradation. The optimal conditions for decolorization with suspended growing cells of strain TL-F1 were as follows: 6-10 g/L sucrose, 5-7 g/L urea, ≥6 % (v/v) inoculation size, ≥160 rpm, 35-40 °C, and pH 5.0-6.0; and those for immobilized cells, the conditions were as follows: 4-6 g/L glucose, 0.2-0.4 g/L urea, 6-10 g/L (wet cell pellets) inoculation size, ≥160 rpm, 35-40 °C, and pH 5.0-7.0. Results of UV-vis scanning spectra suggested that AOG was decolorized through biodegradation, and the possible pathway was proposed through the results of HPLC-MS analysis and related literature. This is a systematic research on aerobic decolorization and degradation of AOG by both suspended and immobilized cells of a C. tropicalis strain.

  10. Biophysical assays to probe the mechanical properties of the interphase cell nucleus: substrate strain application and microneedle manipulation.

    Science.gov (United States)

    Lombardi, Maria L; Zwerger, Monika; Lammerding, Jan

    2011-09-14

    In most eukaryotic cells, the nucleus is the largest organelle and is typically 2 to 10 times stiffer than the surrounding cytoskeleton; consequently, the physical properties of the nucleus contribute significantly to the overall biomechanical behavior of cells under physiological and pathological conditions. For example, in migrating neutrophils and invading cancer cells, nuclear stiffness can pose a major obstacle during extravasation or passage through narrow spaces within tissues.(1) On the other hand, the nucleus of cells in mechanically active tissue such as muscle requires sufficient structural support to withstand repetitive mechanical stress. Importantly, the nucleus is tightly integrated into the cellular architecture; it is physically connected to the surrounding cytoskeleton, which is a critical requirement for the intracellular movement and positioning of the nucleus, for example, in polarized cells, synaptic nuclei at neuromuscular junctions, or in migrating cells.(2) Not surprisingly, mutations in nuclear envelope proteins such as lamins and nesprins, which play a critical role in determining nuclear stiffness and nucleo-cytoskeletal coupling, have been shown recently to result in a number of human diseases, including Emery-Dreifuss muscular dystrophy, limb-girdle muscular dystrophy, and dilated cardiomyopathy.(3) To investigate the biophysical function of diverse nuclear envelope proteins and the effect of specific mutations, we have developed experimental methods to study the physical properties of the nucleus in single, living cells subjected to global or localized mechanical perturbation. Measuring induced nuclear deformations in response to precisely applied substrate strain application yields important information on the deformability of the nucleus and allows quantitative comparison between different mutations or cell lines deficient for specific nuclear envelope proteins. Localized cytoskeletal strain application with a microneedle is used to

  11. Characterization of Strain Pseudomonas sp.Q1 in Microbial Fuel Cell for Treatment of Quinoline-Contaminated Water

    Institute of Scientific and Technical Information of China (English)

    ZHANG Cui-Ping; CHEN Shan-Shan; LIU Guang-Li; ZHANG Ren-Duo; XIE Jian

    2012-01-01

    To find new strain in the microbial fuel cell (MFC) for quinoline removal from wastewater and soil,a facultative anaerobic bacterium strain was isolated from the anode of MFC,utilizing quinoline as the carbon source and electron donor.Based on the 16S rRNA sequence analysis,the bacterium strain was Gram-negative and identified as Pseudomonas sp.Q1 according to its morphology and physiochemical properties.The strain was inoculated into a double-chambered MFC using various quinoline concentratious (0,50,75,86,100,150,200 and 300 mg L-1) combining with 300 mg L-1 glucose as the fuel.Results showed that electricity was generated from the MFC,in which quinoline was degraded simultaneously.The values of Coulombic efficiency (CE) increased with the increase of quinoline concentrations from 0 to 100 mg L-1 then decreased with the increase of quinoline concentration from 100 to 300 mg L-1,and the maximum CE 36.7% was obtained at the quinoline concentration of 100 mg L-1.The cyclic voltammetry analysis suggested that the mechanism of electron transfer was through excreting mediators produced by the strain Q1.The MFC should be a potential method for the treatment of quinoline-contaminated water and soil.

  12. Morphological and molecular characterization of new Drosophila cell lines established from a strain permissive for gypsy transposition.

    Science.gov (United States)

    Chalvet, F; Debec, A; Marcaillou, C; Rougeau, C; Bucheton, A

    1998-01-01

    The gypsy element of Drosophila melanogaster is the first retrovirus identified in invertebrates. Its transposition is controlled by a host gene called flamenco (flam): restrictive alleles of this gene maintain the retrovirus in a repressed state while permissive alleles allow high levels of transposition. To develop a cell system to study the gypsy element, we established four independent cell lines derived from the Drosophila strain SS, which contains a permissive allele of flamenco, and which is devoid of transposing copies of gypsy. The ultrastructural analysis of three SS cell lines revealed some remarkable characteristics, such as many nuclear virus-like particles, cytoplasmic dense particles, and massive cisternae filled with a fibrous material of unknown origin. Gypsy intragenomic distribution has been compared between the three cell lines and the original SS fly strain, and revealed in two of the cell lines an increase in copy number of a restriction fragment usually present in active gypsy elements. This multiplication seems to have occurred during the passage to the cell culture. Availability of SS cell lines should assist studies of gypsy transposition and infectivity and might be useful to produce high amounts of gypsy viral particles. These new lines already allowed us to show that the Envelope-like products of gypsy can be expressed as membrane proteins.

  13. Free mycolic acid accumulation in the cell wall of the mce1 operon mutant strain of Mycobacterium tuberculosis.

    Science.gov (United States)

    Cantrell, Sally A; Leavell, Michael D; Marjanovic, Olivera; Iavarone, Anthony T; Leary, Julie A; Riley, Lee W

    2013-10-01

    The lipid-rich cell wall of Mycobacterium tuberculosis, the agent of tuberculosis, serves as an effective barrier against many chemotherapeutic agents and toxic host cell effector molecules, and it may contribute to the mechanism of persistence. Mycobacterium tuberculosis strains mutated in a 13-gene operon called mce1, which encodes a putative ABC lipid transporter, induce aberrant granulomatous response in mouse lungs. Because of the postulated role of the mce1 operon in lipid importation, we compared the cell wall lipid composition of wild type and mce1 operon mutant M. tuberculosis H37Rv strains. High resolution mass spectrometric analyses of the mce1 mutant lipid extracts showed unbound mycolic acids to accumulate in the cell wall. Quantitative analysis revealed a 10.7 fold greater amount of free mycolates in the mutant compared to that of the wild type strain. The free mycolates were comprised of alpha, methoxy and keto mycolates in the ratio 1:0.9:0.6, respectively. Since the mce1 operon is regulated in vivo, the free mycolates that accumulate during infection may serve as a barrier for M. tuberculosis against toxic products and contribute to the pathogen's persistence.

  14. The CXC chemokine stromal cell-derived factor 1 is not responsible for CD8+ T cell suppression of syncytia-inducing strains of HIV-1

    OpenAIRE

    1997-01-01

    Primary CD8+ T cells from HIV+ asymptomatics can suppress virus production from CD4+ T cells acutely infected with either non-syncytia-inducing (NSI) or syncytia-inducing (SI) HIV-1 isolates. NSI strains of HIV-1 predominantly use the CCR5 chemokine receptor as a fusion cofactor, whereas fusion of T cell line-adapted SI isolates is mediated by another chemokine receptor, CXCR4. The CCR5 ligands RANTES (regulated on activation, normal T cell expressed and secreted), macrophage inflammatory pro...

  15. Immunobiotic Lactobacillus rhamnosus strains differentially modulate antiviral immune response in porcine intestinal epithelial and antigen presenting cells

    Science.gov (United States)

    2014-01-01

    Background Previous findings suggested that Lactobacillus rhamnosus CRL1505 is able to increase resistance of children to intestinal viral infections. However, the intestinal cells, cytokines and receptors involved in the immunoregulatory effect of this probiotic strain have not been fully characterized. Results We aimed to gain insight into the mechanisms involved in the immunomodulatory effect of the CRL1505 strain and therefore evaluated in vitro the crosstalk between L. rhamnosus CRL1505, porcine intestinal epithelial cells (IECs) and antigen presenting cells (APCs) from swine Peyer’s patches in order to deepen our knowledge about the mechanisms, through which this strain may help preventing viral diarrhoea episodes. L. rhamnosus CRL1505 was able to induce IFN–α and –β in IECs and improve the production of type I IFNs in response to poly(I:C) challenge independently of Toll-like receptor (TLR)-2 or TLR9 signalling. In addition, the CRL1505 strain induced mRNA expression of IL-6 and TNF-α via TLR2 in IECs. Furthermore, the strain significantly increased surface molecules expression and cytokine production in intestinal APCs. The improved Th1 response induced by L. rhamnosus CRL1505 was triggered by TLR2 signalling and included augmented expression of MHC-II and co-stimulatory molecules and expression of IL-1β, IL-6, and IFN-γ in APCs. IL-10 was also significantly up-regulated by CRL1505 in APCs. Conclusions It was recently reviewed the emergence of TLR agonists as new ways to transform antiviral treatments by introducing panviral therapeutics with less adverse effects than IFN therapies. The use of L. rhamnosus CRL1505 as modulator of innate immunity and inductor of antiviral type I IFNs, IFN-γ, and regulatory IL-10 clearly offers the potential to overcome this challenge. PMID:24886142

  16. Expression, purification, and improved antigenic specificity of a truncated recombinant bp26 protein of Brucella melitensis M5-90: a potential antigen for differential serodiagnosis of brucellosis in sheep and goats.

    Science.gov (United States)

    Liu, Wen-xing; Hu, Sen; Qiao, Zu-jian; Chen, Wei-ye; Liu, Lin-tao; Wang, Fang-kun; Hua, Rong-hong; Bu, Zhi-gao; Li, Xiang-rui

    2011-01-01

    Antibodies produced in animals vaccinated using live attenuated vaccines against Brucella spp. are indistinguishable using current conventional serological tests from those produced in infected animals. One potential approach is to develop marker vaccines in which specific genes have been deleted from parental vaccine strains that show good immunogenicity and vaccine efficacy. Corresponding methods of detection for antibodies raised by the marker vaccine should also be developed. A specific fragment of the bp26 gene of Brucella melitensis M5-90 was cloned into vector pQE32 to construct the recombinant plasmid (pQE32-rΔbp26). It was used to transform Escherichia coli M15 (pREP4) host cells, which expressed the rΔbp26 protein. Subsequently, the recombinant protein was purified by immobilized metal affinity chromatography and size-exclusion chromatography. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the purified rΔbp26 protein was represented by only one band, with a molecular weight of 14 kDa, and it showed good antigenic specificity on western blot and enzyme-linked immunosorbent assay (ELISA). The purified rΔbp26 protein was intended to be used as an antigen to develop a novel ELISA to differentiate animals vaccinated with bp26 mutants of Brucella spp. from those infected naturally and those vaccinated with the parental vaccine strains.

  17. Lactic Acid Bacteria Strains Exert Immunostimulatory Effect on H. pylori-Induced Dendritic Cells

    Directory of Open Access Journals (Sweden)

    Małgorzata Wiese

    2015-01-01

    Full Text Available The aim of this study was to find out if selected lactic acid bacteria (LAB strains (antagonistic or nonantagonistic against H. pylori in vitro would differ in their abilities to modulate the DCs maturation profiles reflected by their phenotype and cytokine expression patterns. Methods. Monocyte-derived DCs maturation was elicited by their direct exposure to the LAB strains of L. rhamnosus 900 or L. paracasei 915 (antagonistic and nonantagonistic to H. pylori, resp., in the presence or absence of H. pylori strain cagA+. The DCs maturation profile was assessed on the basis of surface markers expression and cytokines production. Results. We observed that the LAB strains and the mixtures of LAB with H. pylori are able to induce mature DCs. At the same time, the L. paracasei 915 leads to high IL-10/IL-12p70 cytokine ratio, in contrast to L. rhamnosus 900. Conclusions. This study showed that the analyzed lactobacilli strains are more potent stimulators of DC maturation than H. pylori. Interestingly from the two chosen LAB strains the antagonistic to H. pylori-L. rhamnosus strain 900 has more proinflammatory and probably antibactericidal properties.

  18. The CXC chemokine stromal cell-derived factor 1 is not responsible for CD8+ T cell suppression of syncytia-inducing strains of HIV-1.

    Science.gov (United States)

    Lacey, S F; McDanal, C B; Horuk, R; Greenberg, M L

    1997-09-02

    Primary CD8+ T cells from HIV+ asymptomatics can suppress virus production from CD4(+) T cells acutely infected with either non-syncytia-inducing (NSI) or syncytia-inducing (SI) HIV-1 isolates. NSI strains of HIV-1 predominantly use the CCR5 chemokine receptor as a fusion cofactor, whereas fusion of T cell line-adapted SI isolates is mediated by another chemokine receptor, CXCR4. The CCR5 ligands RANTES (regulated on activation, normal T cell expressed and secreted), macrophage inflammatory protein 1alpha (MIP-1alpha), and MIP-1beta are HIV-1 suppressive factors secreted by CD8+ cells that inhibit NSI viruses. Recently, the CXC chemokine stromal cell-derived factor 1 (SDF-1) was identified as a ligand for CXCR4 and shown to inhibit SI strains. We speculated that SDF-1 might be an effector molecule for CD8+ suppression of SI isolates and assessed several SDF-1 preparations for inhibition of HIV-1LAI-mediated cell-cell fusion, and examined levels of SDF-1 transcripts in CD8(+) T cells. SDF-1 fusion inhibitory activity correlated with the N terminus, and the alpha and beta forms of SDF-1 exhibited equivalent fusion blocking activity. SDF-1 preparations having the N terminus described by Bleul et al. (Bleul, C.C., Fuhlbrigge, R.C., Casasnovas, J.M., Aiuti, A. & Springer, T.A. (1996) J. Exp. Med. 184, 1101-1109) readily blocked HIV-1LAI-mediated fusion, whereas forms containing two or three additional N-terminal amino acids lacked this activity despite their ability to bind and/or signal through CXCR4. Though SDF-1 is constitutively expressed in most tissues, CD8 T cells contained extremely low levels of SDF-1 mRNA transcripts (suppressive activity. We conclude that suppression of SI strains of HIV-1 by CD8+ T cells is unlikely to involve SDF-1.

  19. Phylogenetic relationships among wine yeast strains based on electrophoretic whole-cell protein patterns.

    Science.gov (United States)

    Guillamón, J M; Querol, A; Jiménez, M; Huerta, T

    1993-04-01

    In the present work, a phylogenetic study based on protein electrophoretic profiles of Saccharomyces strains isolated from different Spanish wine regions has been carried out. Qualitative differences between the protein electrophoregrams were found at inter- and intraspecific level, but not between electrophoregrams of strains isolated at the same ecosystem. The numerical analysis of these results allowed us to conclude that intraspecific relationships are determined by ecological factors, as well as human influences (dispersion and artificial selection). A correlation between ecological and/or geographical origin and the relationships among strains was observed.

  20. Disruption of colonic barrier function and induction of mediator release by strains of Campylobacter jejuni that invade epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Johannes Beltinger; Jo del Buono; Maeve M Skelly; John Thornley; Robin C Spiller; William A Stack; Christopher J Hawkey

    2008-01-01

    AIM:To study the mechanisms by which Campylobacter jejuni (C.jejuni) causes inflammation and diarrhea.In particular,direct interactions with intestinal epithelial cells and effects on barrier function are poorly understood.METHODS:To model the initial pathogenic effects of C.jejuni on intestinal epithelium,polarized human colonic HCA-7 monolayerswere grown on permeabilized filters and infected apically with clinical isolates of C.jejuni.Integrity of the monolayer was monitored by changes in monolayer resistance,release of lactate dehydrogenase,mannitol fluxes and electron microscopy.Invasion of HCA-7 cells was assessed by a modified gentamicin protection assay,translocation by counting colony forming units in the basal chamber,stimulation of mediator release by immunoassays and secretory responses in monolayers stimulated by bradykinin in an Ussing chamber.RESULTS:All strains translocated across monolayers but only a minority invaded HCA-7 cells.Strains that invaded HCA-7 cells destroyed rnonolayer resistance over 6 h,accompanied by increased release of lactate dehydrogenase,a four-fold increase in permeability to [3H] mannitol,and ultrastructural disruption of tight junctions,with rounding and lifting of cells off the filter membrane.Synthesis of interleukin (IL)-8 and prostaglandin E2 was increased with strains that invaded the rnonolayer but not with those that did not.CONCLUSION:These data demonstrate two distinct effects of C.jejuni on colonic epithelial cells and provide an informative model for further investigation of initial host cell responses to C.jejuni.

  1. Neutrino and axion bounds from the globular cluster M5 (NGC 5904).

    Science.gov (United States)

    Viaux, N; Catelan, M; Stetson, P B; Raffelt, G G; Redondo, J; Valcarce, A A R; Weiss, A

    2013-12-06

    The red-giant branch (RGB) in globular clusters is extended to larger brightness if the degenerate helium core loses too much energy in "dark channels." Based on a large set of archival observations, we provide high-precision photometry for the Galactic globular cluster M5 (NGC 5904), allowing for a detailed comparison between the observed tip of the RGB with predictions based on contemporary stellar evolution theory. In particular, we derive 95% confidence limits of g(ae)axion-electron coupling and μ(ν)<4.5×10(-12)μ(B) (Bohr magneton μ(B)=e/2m(e)) on a neutrino dipole moment, based on a detailed analysis of statistical and systematic uncertainties. The cluster distance is the single largest source of uncertainty and can be improved in the future.

  2. Higher Derivative Corrections and Central Charges from Wrapped M5-branes

    CERN Document Server

    Baggio, Marco; Mayerson, Daniel R; Robbins, Daniel; Wecht, Brian

    2014-01-01

    We compute four-derivative corrections to the AdS supergravity actions arising from the near-horizon geometry of N M5-branes wrapped on either one or two Riemann surfaces. This setup features the novel presence of both gauged isometries as well as nontrivial hypermultiplets. We argue that the 5d Chern-Simons terms receive not only higher-derivative corrections but also contributions from Killing vector parameters, which we find must also be corrected. We check the central charges found by our supergravity methods against the dual field theory results and find perfect agreement at leading and subleading order in N. Along the way, we find higher derivative corrections to general AdS_5 and AdS_3x\\Sigma_g geometries.

  3. Witten indices of abelian M5 brane on $\\mathbb{R}\\times S^5$

    CERN Document Server

    Bak, Dongsu

    2016-01-01

    Witten indices and partition functions are computed for abelian 6d tensor and hypermultiplets on $\\mathbb{R}\\times S^5$ in Lorentzian signature in an R gauge field background which preserves some supersymmetry. We consider a generic supersymmetric squashing that also admits squashing of the Hopf fiber. Wick rotation to Euclidean M5 brane amounts to Wick rotation of squashing parameters and the hypermultiplet mass parameter. We compute Casimir energies for tensor and hypermultiplets separately for general squashing, and match these with the corresponding gravitational anomaly polynomials. We extract Witten indices on $\\mathbb{R}\\times \\mathbb{CP}^2$ and find that this is zero, again matching with the vanishing anomaly polynomial on an odd dimensional space.

  4. Geomechanical modeling of the nucleation process of Australia's 1989 M5.6 Newcastle earthquake

    Science.gov (United States)

    Klose, Christian D.

    2007-04-01

    Inherent to black-coal mining in New South Wales (Australia) since 1801, the discharge of ground water may have triggered the M5.6 Newcastle earthquake in 1989. 4-dimensional geomechanical model simulations reveal that widespread water removal and coal as deep as a 500 m depth resulted in an unload of the Earth's crust. This unload caused a destabilization process of the pre-existing Newcastle fault in the interior of the crust beneath the Newcastle coal field. In tandem, an increase in shear stress and a decrease in normal stress may have reactivated this reverse fault. Over the course of the last fifty years, elevated levels of lithostatic stress alterations have accelerated. In 1991, based on the modeling of the crust's elastostatic response to the unload, there has been the minimal critical shear stress changes of 0.01 Mega Pascal (0.1 bar) that reached the Newcastle fault at a depth where the 1989 mainshock nucleated. Hence, it can be anticipated that other faults might also be critically stressed in that region for a couple of reasons. First, the size of the area (volume) that is affected by the induced stress changes is larger than the ruptured area of the Newcastle fault. Second, the seismic moment magnitude of the 1989 M5.6 Newcastle earthquake is associated with only a fraction of mass removal (1 of 55), following McGarr's mass-moment relationship. Lastly, these findings confirm ongoing seismicity in the Newcastle region since the beginning of the 19th century after a dormant period of 10,000 years of no seismicity.

  5. Effect of Agave tequilana juice on cell wall polysaccharides of three Saccharomyces cerevisiae strains from different origins.

    Science.gov (United States)

    Aguilar-Uscanga, Blanca; Arrizon, Javier; Ramirez, Jesús; Solis-Pacheco, Josué

    2007-02-01

    In this study, a characterization of cell wall polysaccharide composition of three yeasts involved in the production of agave distilled beverages was performed. The three yeast strains were isolated from different media (tequila, mezcal and bakery) and were evaluated for the beta(1,3)-glucanase lytic activity and the beta-glucan/ mannan ratio during the fermentation of Agave tequilana juice and in YPD media (control). Fermentations were performed in shake flasks with 30 g l(-1) sugar concentration of A. tequilana juice and with the control YPD using 30 g l(-1) of glucose. The three yeasts strains showed different levels of beta-glucan and mannan when they were grown in A. tequilana juice in comparison to the YPD media. The maximum rate of cell wall lyses was 50% lower in fermentations with A. tequilana juice for yeasts isolated from tequila and mezcal than compared to the bakery yeast.

  6. High-level ethanol production from starch by a flocculent Saccharomyces cerevisiae strain displaying cell-surface glucoamylase

    Energy Technology Data Exchange (ETDEWEB)

    Kondo, A.; Shigechi, H.; Abe, M.; Uyama, K. [Dept. of Chemical Science and Engineering, Kobe Univ., Nadaku, Kobe (Japan); Matsumoto, T.; Fukuda, H. [Div. of Molecular Science, Kobe Univ., Nadaku, Kobe (Japan); Takahashi, S.; Ueda, M.; Tanaka, A. [Dept. of Synthetic Chemistry and Biological Chemistry, Kyoto Univ., Sakyoku, Kyoto (Japan); Kishimoto, M. [Dept. of Biotechnology, Osaka Univ., Osaka (Japan)

    2002-07-01

    A Strain of host yeast YF207, which is a tryptophan auxotroph and shows strong flocculation ability, was obtained from Saccharomyces diastaticus ATCC60712 and S. cerevisiae W303-1B by tetrad analysis. The plasmid pGA11, which is a multicopy plasmid for cell-surface expression of the Rhyzopus oryzae glucoamylase/{alpha}-agglutinin fusion protein, was then introduced into this flocculent yeast strain (YF207/pGA11). Yeast YF207/pGA11 grew rapidly under aerobic condition (dissolved oxygen 2.0 ppm), using soluble starch. The harvested cells were used for batch fermentation of soluble starch to ethanol under anaerobic condition and showed high ethanol production rates (0.71 g h{sup -1} I{sup -1}) without a time lag, because glucoamylase was immobilized on the yeast cell surface. During repeated utilization of cells for fermentation, YF207/pGA11 maintained high ethanol production rates over 300 h. Moreover, in fed-batch fermentation with YF207/pGA11 for approximately 120 h, the ethanol concentration reached up to 50 g I{sup -1}. In conclusion, flocculent yeast cells displaying cell-surface glucoamylase are considered to be very effective for the direct fermentation of soluble starch to ethanol. (orig.)

  7. Inhibitory and Apoptosis-Inducing Effects of Newcastle Disease Virus Strain AF2240 on Mammary Carcinoma Cell Line

    Directory of Open Access Journals (Sweden)

    Umar Ahmad

    2015-01-01

    Full Text Available Breast cancer is the malignant tumour that developed from cells of the breast and is the first leading cause of cancer death among women worldwide. Surgery, radiotherapy, and chemotherapy are the available treatments for breast cancer, but these were reported to have side effects. Newcastle disease virus (NDV known as Avian paramyxovirus type-1 (APMV1 belongs to the genus Avulavirus in a family Paramyxoviridae. NDV is shown to be a promising anticancer agent, killing tumour cells while sparing normal cells unharmed. In this study, the oncolytic and cytotoxic activities of NDV AF2240 strain were evaluated on MDA-MB-231, human mammary carcinoma cell line, using MTT assay, and its inhibitory effects were further studied using proliferation and migration assays. Morphological and apoptotic-inducing effects of NDV on MD-MB-231 cells were observed using phase contrast and fluorescence microscopes. Detection of DNA fragmentation was done following terminal deoxyribonucleotide transferase-mediated Br-dUTP nick end labeling staining (TUNEL assay, which confirmed that the mode of death was through apoptosis and was quantified by flow cytometry. Furthermore, analysis of cellular DNA content demonstrated that the virus caused an increase in the sub-G1 phase (apoptotic peak of the cell cycle. It appears that NDV AF2240 strain is a potent anticancer agent that induced apoptosis in time-dependent manner.

  8. The role of CD4 T cell memory in generating protective immunity to novel and potentially pandemic strains of influenza

    Directory of Open Access Journals (Sweden)

    Anthony eDiPiazza

    2016-01-01

    Full Text Available Recent events have made it clear that potentially pandemic strains of influenza regularly pose a threat to human populations. Therefore, it is essential that we develop better strategies to enhance vaccine design and evaluation, to predict those that will be poor responders to vaccination and to identify those that are at particular risk of disease-associated complications following infection. Simplified animal models have revealed the discrete functions that CD4 T cells play in the developing immune response and to influenza immunity. However, humans have a complex immunological history with influenza through periodic infection and vaccination with seasonal variants, leading to the establishment of heterogeneous memory populations of CD4 T cells that participate in subsequent responses. The continual evolution of the influenza-specific CD4 T cell repertoire involves both specificity and function and overlays other restrictions on CD4 T cell activity derived from viral antigen handling and MHC class II:peptide epitope display. Together, these complexities in the influenza-specific CD4 T cell repertoire constitute a formidable obstacle to predicting protective immune response to potentially pandemic strains of influenza and in devising optimal vaccine strategies to potentiate these responses. We suggest that more precise efforts to identify and enumerate both the positive and negative contributors within the CD4 T cell compartment will aid significantly in achievement of these goals.

  9. Invasion and metastasis ability of renal cancer cell strains 786-0: under the influence of miR-141.

    Science.gov (United States)

    Xu, Y; Lv, L N; Guo, Z Y; Zhang, W

    2016-01-01

    This study aimed to explore the invasion and metastasis ability of miR-141 in 786-0 renal cancer tissue cells, as well as identify the key function of endogenous miR-141 in adjustment and control of malignant activities of renal cancer. The renal cancer cell strain with overexpression of miR-141 and its control renal cancer cell line were constructed; methyl thiazolyl tetrazolium (MTT) assay was adopted to measure proliferation of renal cancer cells; Transwell assay was performed to measure the invasion and metastasis ability of cells; MTT assay and fluorescence activated cell sorting (FACS) were used for measurement of cell apoptosis and drug susceptibility. Results indicated that the expression of miR-141 in 786-0 cells could be significantly increased 400-fold by slow viruses that contained miR-141; moreover, c omprehensive functions showed that miR-141 inhibited the invasion and metastasis ability of renal cancer cells to a great extent (p less than 0.001), partially inhibited cell growth (p less than 0.05) and also induced cell cycle to be arrested in G0/G1 as well as reducing the number of cells in S phase (DNA replicative phase). Moreover, miR-141 could not induce morphologic changes of renal cancer cells, had no direct stimulating effect on cell apoptosis and could not improve the drug susceptibility of renal cancer cells to drugs such as cis-Dichlorodiamineplatinum (DDP), 5-fluorouracil (5-FU) and tumor-related apoptosis-inducing ligand (TRAIL). In conclusion, miR-141 can be considered an important cancer suppressor gene of renal cancer by inhibiting proliferation and metastasis of renal cancer cells.

  10. White and red blood cell picture in rabbits experimentally infected with strains of the rabbit haemorrhagic disease (RHD) virus without or with variable haemagglutination capacity.

    Science.gov (United States)

    Niedźwiedzka-Rystwej, P; Tokarz-Deptuła, B; Deptuła, W

    2016-12-01

    The aim of the study was to establish if haemagglutination of rabbit haemorrhagic disease virus (RHDV) affects haematological picture of peripheral blood in rabbits and the pathogenicity of the virus. The study analyzed white and red blood cell picture in rabbits experimentally infected with two non-haemagglutinating (HA-) RHDV strains (Frankfurt and Asturias) and one strain with variable haemagglutination capacity (HA+/-) (Hagenow). Studies with HA- and HA +/- are rare and relate only to 4 HA- strains (2 RHDV: BLA and Rainham; 2 RHDVa: Pv97 and 9905) and 1 HA+/- RHDV strain: ŻD, where less changes in haematological indices and less pathogenicity were observed. We found that changes caused by HA- Frankfurt strain were related to the number of neutrophils and thrombocytes, while in HA- strain Asturias, in thrombocytes and leukocytes. Changes evoked by HA+/- Hagenow strain pertained to the number of eosinophils, thrombocytes, leukocytes, monocytes, and concentration of hemoglobin. Mortality caused by the Frankfurt strain was 100% between 36 and 48 h post infection (p.i.), while that caused by Asturias strain was 100% between 24 and 36 h p.i., and that observed in case of Hagenow strain was 90% between 36 and 48 h p.i. The changes in haematological picture caused by the HA- and HA+/- RHDV strains were less intensive than those found in case of the HA+ RHDV strains, which cannot be confirmed for pathogenicity, and is not in line with the existing hypothesis suggesting higher pathogenicity in HA+ viruses.

  11. Nonbioluminescent strains of Photobacterium phosphoreum produce the cell-to-cell communication signal N-(3-Hydroxyoctanoyl)homoserine lactone

    DEFF Research Database (Denmark)

    Flodgaard, Lars; Dalgaard, Paw; Andersen, Jens Bo

    2005-01-01

    in the AHL monitor, whereas 48 of 103 nonbioluminescent strains did produce AHLs. AHLs were also detected in Aeromonas spp., but not in Shewanella strains. Thin-layer chromatographic profiles of cod extracts and P. phosphoreum culture supernatants identified a molecule similar in relative mobility (R-f value......) and shape to N-(3-hydroxyoctanoyl)homoserine lactone, and the presence of this molecule in culture supernatants from a nonbioluminescent strain of P. phosphoreum was confirmed by high-performance liquid chromatography-positive electrospray high-resolution mass spectrometry. Bioluminescence (in a non...

  12. Characterization of Aeromonas strains isolated from Indian foods using rpoD gene sequencing and whole cell protein analysis.

    Science.gov (United States)

    Nagar, Vandan; Shashidhar, Ravindranath; Bandekar, Jayant R

    2013-04-01

    Aeromonas are responsible for causing gastroenteritis and extra-intestinal infections in humans. Twenty-two Aeromonas strains isolated from different food sources were re-identified up to species level using rpoD gene sequence analysis. Biochemical tests and 16S rRNA gene sequencing were insufficient to identify Aeromonas till species level. However, incorporation of additional biochemical tests lead to correct identification of 95.5 % strains up to species level. The 16S rRNA gene sequencing was useful to identify Aeromonas isolates at the genus level only. Sequences of the rpoD gene showed greater discriminatory power than 16S rRNA gene and provided conclusive discrimination of the strains for which the phenotypic species identification was uncertain. All these 22 strains were accurately identified up to species level by rpoD gene as A. salmonicida (6), A. veronii bv. veronii (4), A. caviae (3), A. hydrophila (2), A. veronii bv. sobria (2), A. jandaei (1), A. trota (1), A. sobria (1), A. allosaccharophila (1) and A. bivalvium (1). All these strains were also characterized using whole cell protein (WCP) analysis by gradient SDS-PAGE and showed different whole cell protein (WCP) profile [22-28 polypeptide bands (~10 to >97 kDa)], indicating high genetic diversity. The present work emphasizes the use of molecular methods such as rpoD gene sequencing along with comprehensive biochemical tests for the rapid and accurate identification of Aeromonas isolates till species level. The WCP profile can be subsequently used to characterize Aeromonas isolates below species level.

  13. Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system

    OpenAIRE

    Fukuda, Takeshi; Tsuchiyama, Kouta; Makishima, Hirokazu; Takayama, Katsumi; Mulchandani, Ashok; Kuroda, Kouichi; Ueda, Mitsuyoshi; Suye, Shin-ichiro

    2010-01-01

    Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30°C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold great...

  14. Nonbioluminescent strains of Photobacterium phosphoreum produce the cell-to-cell communication signal N-(3-Hydroxyoctanoyl)homoserine lactone

    DEFF Research Database (Denmark)

    Flodgaard, Lars; Dalgaard, Paw; Andersen, Jens Bo

    2005-01-01

    Bioluminescence is a common phenotype in marine bacteria, such As Vibrio and Photobacterium species, and can be quorum regulated by N-acylated homoserine lactones (AHLs). We extracted a molecule that induced a bacterial AHL monitor (Agrobacterium tumefaciens NT1 [pZLR4]) from packed cod fillets......, which spoil due to growth of Photobacterium phosphoreum. Interestingly, AHLs were produced by 13 nonbioluminescent strains of P. phosphoreum isolated from the product. Of 177 strains of P. phosphoreum (including 18 isolates from this study), none of 74 bioluminescent strains elicited a reaction......) and shape to N-(3-hydroxyoctanoyl)homoserine lactone, and the presence of this molecule in culture supernatants from a nonbioluminescent strain of P. phosphoreum was confirmed by high-performance liquid chromatography-positive electrospray high-resolution mass spectrometry. Bioluminescence (in a non...

  15. Parasporin-2 from a New Bacillus thuringiensis 4R2 Strain Induces Caspases Activation and Apoptosis in Human Cancer Cells

    Science.gov (United States)

    Asselin, Eric; Parent, Sophie; Côté, Jean-Charles; Sirois, Marc

    2015-01-01

    In previous studies, parasporin-2Aa1, originally isolated from Bacillus thuringiensis strain A1547, was shown to be cytotoxic against specific human cancer cells but the mechanisms of action were not studied. In the present study, we found that proteinase K activated parasporin-2Aa1 protein isolated from a novel B. thuringiensis strain, 4R2, was specifically cytotoxic to endometrial, colon, liver, cervix, breast and prostate cancer. It showed no toxicity against normal cells. Upon treatment with proteinase K-activated parasporin-2Aa1, morphological changes were observed and western blot analysis revealed the cleavage of poly (ADP-Ribose) polymerase, caspase-3 and caspase-9 in cancer cell lines exclusively, indicative of programmed cell death, apoptosis. Flow cytometry analyses,using propidium iodide and annexin V, as well as a caspases 3/7 assay confirmed apoptosis induction. Further analyses were performed to study survival pathways, including AKT, XIAP, ERK1/2 and PAR-4, a known inducer of apoptosis. These results indicate that parasporin-2Aa1 is a selective cytotoxic protein that induces apoptosis in various human cancer cell lines from diverse tissues. PMID:26263002

  16. Modified nucleotides m2G966/m5C967 of Escherichia coli 16S rRNA are required for attenuation of tryptophan operon

    Science.gov (United States)

    Prokhorova, Irina V.; Osterman, Ilya A.; Burakovsky, Dmitry E.; Serebryakova, Marina V.; Galyamina, Maria A.; Pobeguts, Olga V.; Altukhov, Ilya; Kovalchuk, Sergey; Alexeev, Dmitry G.; Govorun, Vadim M.; Bogdanov, Alexey A.; Sergiev, Petr V.; Dontsova, Olga A.

    2013-11-01

    Ribosomes contain a number of modifications in rRNA, the function of which is unclear. Here we show - using proteomic analysis and dual fluorescence reporter in vivo assays - that m2G966 and m5C967 in 16S rRNA of Escherichia coli ribosomes are necessary for correct attenuation of tryptophan (trp) operon. Expression of trp operon is upregulated in the strain where RsmD and RsmB methyltransferases were deleted, which results in the lack of m2G966 and m5C967 modifications. The upregulation requires the trpL attenuator, but is independent of the promotor of trp operon, ribosome binding site of the trpE gene, which follows trp attenuator and even Trp codons in the trpL sequence. Suboptimal translation initiation efficiency in the rsmB/rsmD knockout strain is likely to cause a delay in translation relative to transcription which causes misregulation of attenuation control of trp operon.

  17. Modified nucleotides m(2)G966/m(5)C967 of Escherichia coli 16S rRNA are required for attenuation of tryptophan operon.

    Science.gov (United States)

    Prokhorova, Irina V; Osterman, Ilya A; Burakovsky, Dmitry E; Serebryakova, Marina V; Galyamina, Maria A; Pobeguts, Olga V; Altukhov, Ilya; Kovalchuk, Sergey; Alexeev, Dmitry G; Govorun, Vadim M; Bogdanov, Alexey A; Sergiev, Petr V; Dontsova, Olga A

    2013-11-18

    Ribosomes contain a number of modifications in rRNA, the function of which is unclear. Here we show--using proteomic analysis and dual fluorescence reporter in vivo assays--that m(2)G966 and m(5)C967 in 16S rRNA of Escherichia coli ribosomes are necessary for correct attenuation of tryptophan (trp) operon. Expression of trp operon is upregulated in the strain where RsmD and RsmB methyltransferases were deleted, which results in the lack of m(2)G966 and m(5)C967 modifications. The upregulation requires the trpL attenuator, but is independent of the promotor of trp operon, ribosome binding site of the trpE gene, which follows trp attenuator and even Trp codons in the trpL sequence. Suboptimal translation initiation efficiency in the rsmB/rsmD knockout strain is likely to cause a delay in translation relative to transcription which causes misregulation of attenuation control of trp operon.

  18. EnVision M5 Venus Orbiter Proposal: Opportunities and Challenges

    Science.gov (United States)

    Ghail, Richard; Wilson, Colin F.; Widemann, Thomas

    2016-10-01

    The core goal of EnVision is to detect activity and measure rates of change on Venus, including geological and geochemical cycles involving the interior, surface and atmosphere.It will observe >20% of the surface with all instruments and will obtain gravity and emissivity data globally. The instrument suite for M5 is under review but will likely comprise the same three instruments as at M4: VenSAR, VEM and SRS.VenSAR. The largest payload instrument is a phased array S-band radar, developed from the UK's low-cost NovaSAR-S instrument optimized for Venus. Use of spacecraft pointing for side-looking, instead of a fixed slant, simplifies the observation strategy to three pairs of ~9 minute/orbit (~36° latitude, ~3800 km) pass-to-pass InSAR swaths, two ~9 minute/orbit multipolar (HH-HV-VV) swaths at lower incidence angle for stereo mapping, two ~3 minute/orbit (~12° latitude, ~1300 km) high resolution swath and 1 to 2 S-band emissivity swaths per day plus 50 km2 ~1 m resolution sliding spotlight images. In addition, InSAR will be acquired along a narrow equatorial strip and across the North Pole to measure variability in the spin rate and axis.VEM. The Venus Emissivity Mapper suite comprises two UV and IR spectrometer channels in addition to the VEM-M IR mapping. A filter array provides wavelength stability and maximizes signal to the focal plane array (FPA). VEM-H is high-resolution, nadir-pointing, infrared spectrometer, the ideal instrument to enable characterization of volcanic plumes released from the surface of Venus by observing SO2, H2O and HDO through the 1 µm, 1.7 µm, and 2-2.3 µm atmospheric windows. Specifically, VEM-H is a redesign of the LNO (Limb, Nadir and Occultation) channel of NOMAD, retaining much heritage from the original with minor modifications to meet the science objectives of the M5 EnVision mission. The third channel, VEM-UV is an upper-atmosphere UV spectrometer dedicated to global SO2 & sulfur cycles.SRS. The Subsurface Radar Sounder

  19. Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system.

    Science.gov (United States)

    Fukuda, Takeshi; Tsuchiyama, Kouta; Makishima, Hirokazu; Takayama, Katsumi; Mulchandani, Ashok; Kuroda, Kouichi; Ueda, Mitsuyoshi; Suye, Shin-ichiro

    2010-05-01

    Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30 degrees C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold greater OPH activity compared with cells engineered using glycosylphosphatidylinositol anchor system, and showed 20-fold greater activity than Escherichia coli using the ice nucleation protein anchor system. These results indicate that Flo1p anchor system is suitable for display of OPH in the cell surface-expression systems.

  20. Differentiating Pseudomonas sp. strain ADP cells in suspensions and biofilms using Raman spectroscopy and scanning electron microscopy.

    Science.gov (United States)

    Henry, Victoria A; Jessop, Julie L P; Peeples, Tonya L

    2017-02-01

    High quality spectra of Pseudomonas sp. strain ADP in the planktonic and biofilm state were obtained using Raman microspectroscopy. These spectra enabled the identification of key differences between free and biofilm cells in the fingerprint region of Raman spectra in the nucleic acid, carbohydrate, and protein regions. Scanning electron microscopy (SEM) enabled detailed visualization of ADP biofilm with confirmation of associated extracellular matrix structure. Following extraction and Raman analysis of extracellular polymeric substances, Raman spectral differences between free and biofilm cells were largely attributed to the contribution of extracellular matrix components produced in mature biofilms. Raman spectroscopy complemented with SEM proves to be useful in distinguishing physiological properties among cells of the same species. Graphical Abstract Raman spectroscopy complemented with SEM proves to be useful in distinguishing physiological properties among cells of the same species.

  1. Variable immune cell frequencies in peripheral blood of LEW.1AR1-iddm rats over time compared to other congenic LEW strains

    Science.gov (United States)

    Arndt, T; Jörns, A; Hedrich, H-J; Lenzen, S; Wedekind, D

    2014-01-01

    The LEW.1AR1-iddm rat is an animal model of human type 1 diabetes (T1D), which arose through a spontaneous mutation within the major histocompatibility complex (MHC)-congenic background strain LEW.1AR1. The LEW.1AR1-iddm rat is characterized by two phenotypes: diabetes development with a diabetes incidence of 60% and a variable T cell frequency in peripheral blood. In this study the immune cell repertoire of LEW.1AR1-iddm rats was analysed over time from days 30 to 90 of life and compared to the background strain LEW.1AR1 and the LEW rat strain as well as the LEW.1WR1 rat strain. The LEW.1AR1-iddm rats are characterized by a high variability of CD3+, CD4+ and CD8+ T cell frequencies in peripheral blood over time, and the frequency is unique for each animal. The variability within the frequencies resulted in changes of the CD4+ : CD8+ T cell ratio. The other three rat strains studied were characterized by a stable but nevertheless strain-specific T cell frequency resulting in a specific CD4+ : CD8+ T cell ratio. The frequency of natural killer (NK) cells and B cells in LEW.1AR1-iddm rats was increased, with a higher variability compared to the other strains. Only monocytes showed no differences in frequency and variability between all strains studied. These variabilities of immune cell frequencies in the LEW.1AR1-iddm rats might lead to imbalances between autoreactive and regulatory T cells in peripheral blood as a prerequisite for diabetes development. PMID:24628466

  2. The Adherent/Invasive Escherichia coli Strain LF82 Invades and Persists in Human Prostate Cell Line RWPE-1, Activating a Strong Inflammatory Response

    Science.gov (United States)

    Aleandri, Marta; Marazzato, Massimiliano; Conte, Antonietta L.; Ambrosi, Cecilia; Nicoletti, Mauro; Zagaglia, Carlo; Gambara, Guido; Palombi, Fioretta; De Cesaris, Paola; Ziparo, Elio; Palamara, Anna T.; Riccioli, Anna

    2016-01-01

    Adherent/invasive Escherichia coli (AIEC) strains have recently been receiving increased attention because they are more prevalent and persistent in the intestine of Crohn's disease (CD) patients than in healthy subjects. Since AIEC strains show a high percentage of similarity to extraintestinal pathogenic E. coli (ExPEC), neonatal meningitis-associated E. coli (NMEC), and uropathogenic E. coli (UPEC) strains, here we compared AIEC strain LF82 with a UPEC isolate (strain EC73) to assess whether LF82 would be able to infect prostate cells as an extraintestinal target. The virulence phenotypes of both strains were determined by using the RWPE-1 prostate cell line. The results obtained indicated that LF82 and EC73 are able to adhere to, invade, and survive within prostate epithelial cells. Invasion was confirmed by immunofluorescence and electron microscopy. Moreover, cytochalasin D and colchicine strongly inhibited bacterial uptake of both strains, indicating the involvement of actin microfilaments and microtubules in host cell invasion. Moreover, both strains belong to phylogenetic group B2 and are strong biofilm producers. In silico analysis reveals that LF82 shares with UPEC strains several virulence factors: namely, type 1 pili, the group II capsule, the vacuolating autotransporter toxin, four iron uptake systems, and the pathogenic island (PAI). Furthermore, compared to EC73, LF82 induces in RWPE-1 cells a marked increase of phosphorylation of mitogen-activated protein kinases (MAPKs) and of NF-κB already by 5 min postinfection, thus inducing a strong inflammatory response. Our in vitro data support the hypothesis that AIEC strains might play a role in prostatitis, and, by exploiting host-cell signaling pathways controlling the innate immune response, likely facilitate bacterial multiplication and dissemination within the male genitourinary tract. PMID:27600504

  3. Observations of static Coulomb stress triggering of the November 2011 M5.7 Oklahoma earthquake sequence

    Science.gov (United States)

    Sumy, Danielle F.; Cochran, Elizabeth S.; Keranen, Katie M.; Wei, Maya; Abers, Geoffrey A.

    2014-01-01

    In November 2011, a M5.0 earthquake occurred less than a day before a M5.7 earthquake near Prague, Oklahoma, which may have promoted failure of the mainshock and thousands of aftershocks along the Wilzetta fault, including a M5.0 aftershock. The M5.0 foreshock occurred in close proximity to active fluid injection wells; fluid injection can cause a buildup of pore fluid pressure, decrease the fault strength, and may induce earthquakes. Keranen et al. [2013] links the M5.0 foreshock with fluid injection, but the relationship between the foreshock and successive events has not been investigated. Here we examine the role of coseismic Coulomb stress transfer on earthquakes that follow the M5.0 foreshock, including the M5.7 mainshock. We resolve the static Coulomb stress change onto the focal mechanism nodal plane that is most consistent with the rupture geometry of the three M ≥ 5.0 earthquakes, as well as specified receiver fault planes that reflect the regional stress orientation. We find that Coulomb stress is increased, e.g., fault failure is promoted, on the nodal planes of ~60% of the events that have focal mechanism solutions, and more specifically, that the M5.0 foreshock promoted failure on the rupture plane of the M5.7 mainshock. We test our results over a range of effective coefficient of friction values. Hence, we argue that the M5.0 foreshock, induced by fluid injection, potentially triggered a cascading failure of earthquakes along the complex Wilzetta fault system.

  4. RR Lyrae stars and the horizontal branch of NGC 5904 (M5)

    CERN Document Server

    Ferro, A Arellano; Bramich, D M; Giridhar, Sunetra; Ahumada, J A; Muneer, S

    2016-01-01

    We report the distance and [Fe/H] value for the globular cluster NGC 5904 (M5) derived from the Fourier decomposition of the light curves of selected RRab and RRc stars. The aim in doing this was to bring these parameters into the homogeneous scales established by our previous work on numerous other globular clusters, allowing a direct comparison of the horizontal branch luminosity in clusters with a wide range of metallicities. Our CCD photometry of the large variable star population of this cluster is used to discuss light curve peculiarities, like Blazhko modulations, on an individual basis. New Blazhko variables are reported. From the RRab stars we found [Fe/H]$_{\\rm UVES} = -1.335 \\pm 0.003{\\rm(statistical)} \\pm 0.110{\\rm(systematic)}$, and a distance of $7.6\\pm 0.2$ kpc, and from the RRc stars we found [Fe/H]$_{\\rm UVES}$ = $-1.39 \\pm 0.03{\\rm(statistical)} \\pm 0.12{\\rm(systematic)}$ and a distance of $7.5 \\pm 0.3$ kpc. The results for RRab and RRc stars should be considered independent since they come ...

  5. Wavelet coupled MARS and M5 Model Tree approaches for groundwater level forecasting

    Science.gov (United States)

    Rezaie-balf, Mohammad; Naganna, Sujay Raghavendra; Ghaemi, Alireza; Deka, Paresh Chandra

    2017-10-01

    In this study, two different machine learning models, Multivariate Adaptive Regression Splines (MARS) and M5 Model Trees (MT) have been applied to simulate the groundwater level (GWL) fluctuations of three shallow open wells within diverse unconfined aquifers. The Wavelet coupled MARS and MT hybrid models were developed in an attempt to further increase the GWL forecast accuracy. The Discrete Wavelet Transform (DWT) which is particularly effective in dealing with non-stationary time-series data was employed to decompose the input time series into various sub-series components. Historical data of 10 years (August-1996 to July-2006) comprising monthly groundwater level, rainfall, and temperature were used to calibrate and validate the models. The models were calibrated and tested for one, three and six months ahead forecast horizons. The wavelet coupled MARS and MT models were compared with their simple counterpart using standard statistical performance evaluation measures such as Root Mean Square Error (RMSE), Normalized Nash-Sutcliffe Efficiency (NNSE) and Coefficient of Determination (R2) . The wavelet coupled MARS and MT models developed using multi-scale input data performed better compared to their simple counterpart and the forecast accuracy of W-MARS models were superior to that of W-MT models. Specifically, the DWT offered a better discrimination of non-linear and non-stationary trends that were present at various scales in the time series of the input variables thus crafting the W-MARS models to provide more accurate GWL forecasts.

  6. Lithium abundances in globular cluster giants: NGC 6218 (M12) and NGC 5904 (M5)

    CERN Document Server

    D'Orazi, Valentina; Gratton, Raffaele G; Lattanzio, John C; Bragaglia, Angela; Carretta, Eugenio; Lucatello, Sara; Momany, Yazan

    2014-01-01

    Convergent lines of evidence suggest that globular clusters host multiple stellar populations. It appears that they experience at least two episodes of star formation whereby a fraction of first-generation stars contribute astrated ejecta to form the second generation(s). To identify the polluting progenitors we require distinguishing chemical signatures such as that provided by lithium. Theoretical models predict that lithium can be synthesised in AGB stars, whereas no net Li production is expected from other candidates. It has been shown that in order to reproduce the abundance pattern found in M4, Li production must occur within the polluters, favouring the AGB scenario. Here we present Li and Al abundances for a large sample of RGB stars in M12 and M5. These clusters have a very similar metallicity, whilst demonstrating differences in several cluster properties. Our results indicate that the first-generation and second-generation stars share the same Li content in M12; we recover an abundance pattern simi...

  7. The intrinsic extreme ultraviolet fluxes of F5 V to M5 V stars

    Energy Technology Data Exchange (ETDEWEB)

    Linsky, Jeffrey L. [JILA, University of Colorado and NIST, 440UCB Boulder, CO 80309-0440 (United States); Fontenla, Juan [NorthWest Research Associates Inc., 3380 Mitchell Ln, Boulder, CO 80301 (United States); France, Kevin, E-mail: jlinsky@jilau1.colorado.edu, E-mail: jfontenla@nwra.com, E-mail: Kevin.France@colorado.edu [CASA, University of Colorado, 593UCB Boulder, CO 80309-0593 (United States)

    2014-01-01

    Extreme ultraviolet (EUV) radiations (10-117 nm) from host stars play important roles in the ionization, heating, and mass loss from exoplanet atmospheres. Together with the host star's Lyα and far-UV (117-170 nm) radiation, EUV radiation photodissociates important molecules, thereby changing the chemistry in exoplanet atmospheres. Since stellar EUV fluxes cannot now be measured and interstellar neutral hydrogen completely obscures stellar radiation between 40 and 91.2 nm, even for the nearest stars, we must estimate the unobservable EUV flux by indirect methods. New non-LTE semiempirical models of the solar chromosphere and corona and solar irradiance measurements show that the ratio of EUV flux in a variety of wavelength bands to the Lyα flux varies slowly with the Lyα flux and thus with the magnetic heating rate. This suggests and we confirm that solar EUV/Lyα flux ratios based on the models and observations are similar to the available 10-40 nm flux ratios observed with the Extreme Ultraviolet Explorer (EUVE) satellite and the 91.2-117 nm flux observed with the Far Ultraviolet Spectroscopic Explorer (FUSE) satellite for F5 V-M5 V stars. We provide formulae for predicting EUV flux ratios based on the EUVE and FUSE stellar data and on the solar models, which are essential input for modeling the atmospheres of exoplanets.

  8. Increasing cell biomass in Saccharomyces cerevisiae increases recombinant protein yield: the use of a respiratory strain as a microbial cell factory

    Directory of Open Access Journals (Sweden)

    Hedfalk Kristina

    2010-06-01

    Full Text Available Abstract Background Recombinant protein production is universally employed as a solution to obtain the milligram to gram quantities of a given protein required for applications as diverse as structural genomics and biopharmaceutical manufacture. Yeast is a well-established recombinant host cell for these purposes. In this study we wanted to investigate whether our respiratory Saccharomyces cerevisiae strain, TM6*, could be used to enhance the productivity of recombinant proteins over that obtained from corresponding wild type, respiro-fermentative strains when cultured under the same laboratory conditions. Results Here we demonstrate at least a doubling in productivity over wild-type strains for three recombinant membrane proteins and one recombinant soluble protein produced in TM6* cells. In all cases, this was attributed to the improved biomass properties of the strain. The yield profile across the growth curve was also more stable than in a wild-type strain, and was not further improved by lowering culture temperatures. This has the added benefit that improved yields can be attained rapidly at the yeast's optimal growth conditions. Importantly, improved productivity could not be reproduced in wild-type strains by culturing them under glucose fed-batch conditions: despite having achieved very similar biomass yields to those achieved by TM6* cultures, the total volumetric yields were not concomitantly increased. Furthermore, the productivity of TM6* was unaffected by growing cultures in the presence of ethanol. These findings support the unique properties of TM6* as a microbial cell factory. Conclusions The accumulation of biomass in yeast cell factories is not necessarily correlated with a proportional increase in the functional yield of the recombinant protein being produced. The respiratory S. cerevisiae strain reported here is therefore a useful addition to the matrix of production hosts currently available as its improved biomass

  9. Integration of a laterally acquired gene into a cell network important for growth in a strain of Vibrio rotiferianus

    Directory of Open Access Journals (Sweden)

    Labbate Maurizio

    2011-11-01

    Full Text Available Abstract Background Lateral Gene Transfer (LGT is a major contributor to bacterial evolution and up to 25% of a bacterium's genome may have been acquired by this process over evolutionary periods of time. Successful LGT requires both the physical transfer of DNA and its successful incorporation into the host cell. One system that contributes to this latter step by site-specific recombination is the integron. Integrons are found in many diverse bacterial Genera and is a genetic system ubiquitous in vibrios that captures mobile DNA at a dedicated site. The presence of integron-associated genes, contained within units of mobile DNA called gene cassettes makes up a substantial component of the vibrio genome (1-3%. Little is known about the role of this system since the vast majority of genes in vibrio arrays are highly novel and functions cannot be ascribed. It is generally regarded that strain-specific mobile genes cannot be readily integrated into the cellular machinery since any perturbation of core metabolism is likely to result in a loss of fitness. Results In this study, at least one mobile gene contained within the Vibrio rotiferianus strain DAT722, but lacking close relatives elsewhere, is shown to greatly reduce host fitness when deleted and tested in growth assays. The precise role of the mobile gene product is unknown but impacts on the regulation of outermembrane porins. This demonstrates that strain specific laterally acquired mobile DNA can be integrated rapidly into bacterial networks such that it becomes advantageous for survival and adaptation in changing environments. Conclusions Mobile genes that are highly strain specific are generally believed to act in isolation. This is because perturbation of existing cell machinery by the acquisition of a new gene by LGT is highly likely to lower fitness. In contrast, we show here that at least one mobile gene, apparently unique to a strain, encodes a product that has integrated into central

  10. Combined effects of proinflammatory cytokines and intermittent cyclic mechanical strain in inhibiting osteogenicity in human periodontal ligament cells.

    Science.gov (United States)

    Sun, Chaofan; Chen, Lijiao; Shi, Xinlian; Cao, Zhensheng; Hu, Bibo; Yu, Wenbin; Ren, Manman; Hu, Rongdang; Deng, Hui

    2016-09-01

    Mechanical strain plays an important role in bone formation and resorption during orthodontic tooth movement. The mechanism has not been fully studied, and the process becomes complex with increased amounts of periodontal patients seeking orthodontic care. Our aims were to elucidate the combined effects of proinflammatory cytokines and intermittent cyclic strain (ICS) on the osteogenic capacity of human periodontal ligament cells. Cultured human periodontal ligament cells were exposed to proinflammatory cytokines (interleukin-1β 5 ng/mL and tumor necrosis factor-α 10 ng/mL) for 1 and 5 days, and ICS (0.5 Hz, 12% elongation) was applied for 4 h per day. The autocrine of inflammatory cytokines was measured by enzyme-linked immunosorbent assay. The expression of osteoblast markers runt-related transcription factor 2 and rabbit collagen type I was determined using real-time polymerase chain reaction and Western blot. The osteogenic capacity was also detected by alkaline phosphatase (ALP) staining, ALP activity, and alizarin red staining. We demonstrated that ICS impaired the osteogenic capacity of human periodontal ligament cells when incubated with proinflammatory cytokines, as evidenced by the low expression of ALP staining, low ALP activity, reduced alizarin red staining, and reduced osteoblast markers. These data, for the first time, suggest that ICS has a negative effect on the inductive inhibition of osteogenicity in human PDL cells mediated by proinflammatory cytokines.

  11. Post-invasion events after infection with Staphylococcus aureus are strongly dependent on both the host cell type and the infecting S. aureus strain.

    Science.gov (United States)

    Strobel, M; Pförtner, H; Tuchscherr, L; Völker, U; Schmidt, F; Kramko, N; Schnittler, H-J; Fraunholz, M J; Löffler, B; Peters, G; Niemann, S

    2016-09-01

    Host cell invasion is a major feature of Staphylococcus aureus and contributes to infection development. The intracellular metabolically active bacteria can induce host cell activation and death but they can also persist for long time periods. In this study a comparative analysis was performed of different well-characterized S. aureus strains in their interaction with a variety of host cell types. Staphylococcus aureus (strains 6850, USA300, LS1, SH1000, Cowan1) invasion was compared in different human cell types (epithelial and endothelial cells, keratinocytes, fibroblasts, osteoblasts). The number of intracellular bacteria was determined, cell inflammation was investigated, as well as cell death and phagosomal escape of bacteria. To explain strain-dependent differences in the secretome, a proteomic approach was used. Barrier cells took up high amounts of bacteria and were killed by aggressive strains. These strains expressed high levels of toxins, and possessed the ability to escape from phagolysosomes. Osteoblasts and keratinocytes ingested less bacteria, and were not killed, even though the primary osteoblasts were strongly activated by S. aureus. In all cell types S. aureus was able to persist. Strong differences in uptake, cytotoxicity, and inflammatory response were observed between primary cells and their corresponding cell lines, demonstrating that cell lines reflect only partially the functions and physiology of primary cells. This study provides a contribution for a better understanding of the pathomechanisms of S. aureus infections. The proteomic data provide important basic knowledge on strains commonly used in the analysis of S. aureus-host cell interaction.

  12. Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications.

    Science.gov (United States)

    Friedrich, Valentin; Janesch, Bettina; Windwarder, Markus; Maresch, Daniel; Braun, Matthias L; Megson, Zoë A; Vinogradov, Evgeny; Goneau, Marie-France; Sharma, Ashu; Altmann, Friedrich; Messner, Paul; Schoenhofen, Ian C; Schäffer, Christina

    2016-12-16

    Tannerella forsythia is an anaerobic, Gram-negative periodontal pathogen. A unique O-linked oligosaccharide decorates the bacterium's cell surface proteins and was shown to modulate the host immune response. In our study, we investigated the biosynthesis of the nonulosonic acid (NulO) present at the terminal position of this glycan. A bioinformatic analysis of T. forsythia genomes revealed a gene locus for the synthesis of pseudaminic acid (Pse) in the type strain ATCC 43037 while strains FDC 92A2 and UB4 possess a locus for the synthesis of legionaminic acid (Leg) instead. In contrast to the NulO in ATCC 43037, which has been previously identified as a Pse derivative (5-N-acetimidoyl-7-N-glyceroyl-3,5,7,9-tetradeoxy-l-glycero-l-manno-NulO), glycan analysis of strain UB4 performed in this study indicated a 350-Da, possibly N-glycolyl Leg (3,5,7,9-tetradeoxy-d-glycero-d-galacto-NulO) derivative with unknown C5,7 N-acyl moieties. We have expressed, purified and characterized enzymes of both NulO pathways to confirm these genes' functions. Using capillary electrophoresis (CE), CE-mass spectrometry and NMR spectroscopy, our studies revealed that Pse biosynthesis in ATCC 43037 essentially follows the UDP-sugar route described in Helicobacter pylori, while the pathway in strain FDC 92A2 corresponds to Leg biosynthesis in Campylobacter jejuni involving GDP-sugar intermediates. To demonstrate that the NulO biosynthesis enzymes are functional in vivo, we created knockout mutants resulting in glycans lacking the respective NulO. Compared to the wild-type strains, the mutants exhibited significantly reduced biofilm formation on mucin-coated surfaces, suggestive of their involvement in host-pathogen interactions or host survival. This study contributes to understanding possible biological roles of bacterial NulOs.

  13. Critical study of the method of calculating virgin rock stresses from measurement results of the CSIR triaxial strain cell

    Science.gov (United States)

    Vreede, F. A.

    1981-05-01

    The manual of instructions for the user of the CSIR triaxial rock stress measuring equipment is critically examined. It is shown that the values of the rock stresses can be obtained from the strain gauge records by means of explicit formulae, which makes the manual's computer program obsolete. Furthermore statistical methods are proposed to check for faulty data and inhomogeneity in rock properties and virgin stress. The possibility of non-elastic behavior of the rock during the test is also checked. A new computer program based on the explicit functions and including the check calculations is presented. It is much more efficient than the one in the manual since it does not require computer sub-routines, allowing it to be used directly on any modern computer. The output of the new program is in a format suitable for direct inclusion in the report of an investigation using strain cell results.

  14. Unequivocal identification of subpopulations in putative multiclonal Trypanosoma cruzi strains by FACs single cell sorting and genotyping.

    Directory of Open Access Journals (Sweden)

    Helder Magno Silva Valadares

    Full Text Available Trypanosoma cruzi, the etiological agent of Chagas disease, is a polymorphic species. Evidence suggests that the majority of the T. cruzi populations isolated from afflicted humans, reservoir animals, or vectors are multiclonal. However, the extent and the complexity of multiclonality remain to be established, since aneuploidy cannot be excluded and current conventional cloning methods cannot identify all the representative clones in an infection. To answer this question, we adapted a methodology originally described for analyzing single spermatozoids, to isolate and study single T. cruzi parasites. Accordingly, the cloning apparatus of a Fluorescence-Activated Cell Sorter (FACS was used to sort single T. cruzi cells directly into 96-wells microplates. Cells were then genotyped using two polymorphic genomic markers and four microsatellite loci. We validated this methodology by testing four T. cruzi populations: one control artificial mixture composed of two monoclonal populations--Silvio X10 cl1 (TcI and Esmeraldo cl3 (TcII--and three naturally occurring strains, one isolated from a vector (A316A R7 and two others derived from the first reported human case of Chagas disease. Using this innovative approach, we were able to successfully describe the whole complexity of these natural strains, revealing their multiclonal status. In addition, our results demonstrate that these T. cruzi populations are formed of more clones than originally expected. The method also permitted estimating of the proportion of each subpopulation of the tested strains. The single-cell genotyping approach allowed analysis of intrapopulation diversity at a level of detail not achieved previously, and may thus improve our comprehension of population structure and dynamics of T. cruzi. Finally, this methodology is capable to settle once and for all controversies on the issue of multiclonality.

  15. Cell surface-associated compounds of probiotic lactobacilli sustain the strain-specificity dogma

    NARCIS (Netherlands)

    Bron, P.A.; Tomita, S.; Mercenier, A.M.E.; Kleerebezem, M.

    2013-01-01

    Probiotic lactobacilli can positively impact on the health status of targeted (diseased) populations but efficacy depends strongly on the strain employed and the molecular basis for this phenomenon is poorly understood. This review discusses the current state-of-the-art in the field of molecular

  16. Cell surface-associated compounds of probiotic lactobacilli sustain the strain-specificity dogma

    NARCIS (Netherlands)

    Bron, P.A.; Tomita, S.; Mercenier, A.M.E.; Kleerebezem, M.

    2013-01-01

    Probiotic lactobacilli can positively impact on the health status of targeted (diseased) populations but efficacy depends strongly on the strain employed and the molecular basis for this phenomenon is poorly understood. This review discusses the current state-of-the-art in the field of molecular pro

  17. Strain Library Imaging Protocol for high-throughput, automated single-cell microscopy of large bacterial collections arrayed on multiwell plates.

    Science.gov (United States)

    Shi, Handuo; Colavin, Alexandre; Lee, Timothy K; Huang, Kerwyn Casey

    2017-02-01

    Single-cell microscopy is a powerful tool for studying gene functions using strain libraries, but it suffers from throughput limitations. Here we describe the Strain Library Imaging Protocol (SLIP), which is a high-throughput, automated microscopy workflow for large strain collections that requires minimal user involvement. SLIP involves transferring arrayed bacterial cultures from multiwell plates onto large agar pads using inexpensive replicator pins and automatically imaging the resulting single cells. The acquired images are subsequently reviewed and analyzed by custom MATLAB scripts that segment single-cell contours and extract quantitative metrics. SLIP yields rich data sets on cell morphology and gene expression that illustrate the function of certain genes and the connections among strains in a library. For a library arrayed on 96-well plates, image acquisition can be completed within 4 min per plate.

  18. Characterization of exoelectrogenic bacteria enterobacter strains isolated from a microbial fuel cell exposed to copper shock load.

    Directory of Open Access Journals (Sweden)

    Cuijie Feng

    Full Text Available Microorganisms capable of generating electricity in microbial fuel cells (MFCs have gained increasing interest. Here fourteen exoelectrogenic bacterial strains were isolated from the anodic biofilm in an MFC before and after copper (Cu shock load by Hungate roll-tube technique with solid ferric (III oxide as an electron acceptor and acetate as an electron donor. Phylogenetic analysis of the 16S rRNA gene sequences revealed that they were all closely related to Enterobacter ludwigii DSM 16688T within the Enterobacteriaceae family, although these isolated bacteria showed slightly different morphology before and after Cu shock load. Two representative strains R2B1 (before Cu shock load and B4B2 (after Cu shock load were chosen for further analysis. B4B2 is resistant to 200 mg L-1 of Cu(II while R2B1 is not, which indicated the potential selection of the Cu shock load. Raman analysis revealed that both R2B1 and B4B2 contained c-type cytochromes. Cyclic voltammetry measurements revealed that strain R2B1 had the capacity to transfer electrons to electrodes. The experimental results demonstrated that strain R2B1 was capable of utilizing a wide range of substrates, including Luria-Bertani (LB broth, cellulose, acetate, citrate, glucose, sucrose, glycerol and lactose to generate electricity, with the highest current density of 440 mA·m-2 generated from LB-fed MFC. Further experiments indicated that the bacterial cell density had potential correlation with the current density.

  19. Novel SLA-DR alleles of three Chinese pig strains and the related function in human T cell response.

    Science.gov (United States)

    Chen, Fuxiang; Xie, Jin; Zhou, Yun; Li, Ningli; Chou, Kuang-Yen

    2004-06-01

    To elucidate the structures of SLA-DR (swine leukocyte antigen DR) genes of three Chinese pig strains (Gz, Bm and Yn), the SLA-DRA and SLA-DRB cDNA were amplified by RT-PCR and subjected to determine the sequences. The whole structures of SLA-DRA alleles are identical among three strains, consisting of 759 nucleotides including an open reading frame (ORF), and are shared with those reported from NIH minipigs SLA-DRA(c) and SLA-DRA(d). The same length of the ORF-containing SLA-DRB genes of three Chinese pig strains was also identified. They are composed of 801 nucleotides encoding a xenogeneic antigen molecule of 266 amino acid residues. The nucleotide sequences of the SLA-DRB genes, however, are different when compared either among the three strains or with the published data of SLA-DRB sequences, which allowed our novel SLA-DRB alleles receiving their accession numbers AY102479, AY102480 and AY102481 from the GenBank. This study further reveals that the phylogenic homologies of MHC DR or DR-like genes in structures of nucleotides and deduced amino acids between Chinese pigs (SLA) and human (HLA-DRB1*0901) are better than those between pigs and mice (H-2(b)Ebeta). High similarities were also found for DRalpha-DRbeta heterodimers between Chinese pigs and human in terms of amino acids sequences critical for binding with human CD4 coreceptor molecule, which are better than those between SLA-DR and H-2 I-E molecules. A functional test indicated that, by cotransfection with Bm-DRA and Bm-DRB genes, the Bm-DR molecule-expressed L929 cells could stimulate human T cells quite well in a xenogeneic reaction in presence of human APCs.

  20. Characterization of exoelectrogenic bacteria enterobacter strains isolated from a microbial fuel cell exposed to copper shock load.

    Science.gov (United States)

    Feng, Cuijie; Li, Jiangwei; Qin, Dan; Chen, Lixiang; Zhao, Feng; Chen, Shaohua; Hu, Hongbo; Yu, Chang-Ping

    2014-01-01

    Microorganisms capable of generating electricity in microbial fuel cells (MFCs) have gained increasing interest. Here fourteen exoelectrogenic bacterial strains were isolated from the anodic biofilm in an MFC before and after copper (Cu) shock load by Hungate roll-tube technique with solid ferric (III) oxide as an electron acceptor and acetate as an electron donor. Phylogenetic analysis of the 16S rRNA gene sequences revealed that they were all closely related to Enterobacter ludwigii DSM 16688T within the Enterobacteriaceae family, although these isolated bacteria showed slightly different morphology before and after Cu shock load. Two representative strains R2B1 (before Cu shock load) and B4B2 (after Cu shock load) were chosen for further analysis. B4B2 is resistant to 200 mg L-1 of Cu(II) while R2B1 is not, which indicated the potential selection of the Cu shock load. Raman analysis revealed that both R2B1 and B4B2 contained c-type cytochromes. Cyclic voltammetry measurements revealed that strain R2B1 had the capacity to transfer electrons to electrodes. The experimental results demonstrated that strain R2B1 was capable of utilizing a wide range of substrates, including Luria-Bertani (LB) broth, cellulose, acetate, citrate, glucose, sucrose, glycerol and lactose to generate electricity, with the highest current density of 440 mA·m-2 generated from LB-fed MFC. Further experiments indicated that the bacterial cell density had potential correlation with the current density.

  1. Novel SLA-DR Alleles of Three Chinese Pig Strains and the Related Function in Human T Cell Response

    Institute of Scientific and Technical Information of China (English)

    FuxiangChen; JinXie; YunZhou; NingliLi; Kuang-YenChou

    2004-01-01

    To elucidate the structures of SLA-DR (swine leukocyte antigen DR) genes of three Chinese pig strains (Gz, Bm and Yn), the SLA-DRA and SLA-DRB cDNA were amplified by RT-PCR and subjected to determine the sequences. The whole structures of SLA-DRA alleles are identical among three strains, consisting of 759 nucleotides including an open reading frame (ORF), and are shared with those reported from NIH minipigs SLA-DRAc and SLA-DRAd. The same length of the ORF-containing SLA-DRB genes of three Chinese pig strains was also identified. They are composed of 801 nucleotides encoding a xenogeneic antigen molecule of 266 amino acid residues. The nucleotide sequences of the SLA-DRB genes, however, are different when compared either among the three strains or with the published data of SLA-DRB sequences, which allowed our novel SLA-DRB alleles receiving their accession numbers AY102479, AY102480 and AY102481 from the GenBank. This study further reveals that the phylogenic homologies of MHC DR or DR-like genes in structures of nucleotides and deduced amino acids between Chinese pigs (SLA) and human (HLA-DRBI*0901) are better than those between pigs and mice (H-2b Eβ). High similarities were also found for DRα-DRβ heterodimers between Chinese pigs and human in terms of amino acids sequences critical for binding with human CD4 coreceptor molecule, which are better than those between SLA-DR and H-2 I-E molecules. A functional test indicated that, by cotransfection with Bm-DRA and Bm-DRB genes, the Bm-DR molecule-expressed L929 cells could stimulate human T cells quite well in a xenogeneic reaction in presence of human APCs.

  2. Novel SLA-DR Alleles of Three Chinese Pig Strains and the Related Function in Human T Cell Response

    Institute of Scientific and Technical Information of China (English)

    Fuxiang Chen; Jin Xie; Yun Zhou; Ningli Li; Kuang-Yen Chou

    2004-01-01

    To elucidate the structures of SLA-DR (swine leukocyte antigen DR) genes of three Chinese pig strains (Gz, Bm and Yn), the SLA-DRA and SLA-DRB cDNA were amplified by RT-PCR and subjected to determine the sequences. The whole structures of SLA-DRA alleles are identical among three strains, consisting of 759 nucleotides including an open reading frame (ORF), and are shared with those reported from NIH minipigs SLA-DRAc and SLA-DRAd. The same length of the ORF-containing SLA-DRB genes of three Chinese pig strains was also identified. They are composed of 801 nucleotides encoding a xenogeneic antigen molecule of 266 amino acid residues. The nucleotide sequences of the SLA-DRB genes, however, are different when compared either among the three strains or with the published data of SLA-DRB sequences, which allowed our novel SLA-DRB alleles receiving their accession numbers AY102479, AY102480 and AY102481 from the GenBank. This study further reveals that the phylogenic homologies of MHC DR or DR-like genes in structures of nucleotides and deduced amino acids between Chinese pigs (SLA) and human (HLA-DRB1*0901) are better than those between pigs and mice (H-2b Eβ). High similarities were also found for DRα-DRβ heterodimers between Chinese pigs and human in terms of amino acids sequences critical for binding with human CD4 coreceptor molecule, which are better than those between SLA-DR and H-2 I-E molecules. A functional test indicated that, by cotransfection with Bm-DRA and Bm-DRB genes, the Bm-DR molecule-expressed L929 cells could stimulate human T cells quite well in a xenogeneic reaction in presence of human APCs.

  3. The galactic globular cluster M5 (NGC 5904 as a particle physics laboratory

    Directory of Open Access Journals (Sweden)

    Redondo J.

    2013-03-01

    Full Text Available Globular clusters have been used for a long time to test stellar evolution theories, and in particular to constrain novel forms of energy loss in low-mass stars. This includes constraints on axion properties, neutrino dipole moments, milli-charged particles, Kaluza-Klein gravitons, and many other phenomena. Depending on their interaction strength, these particles can be abundantly produced in stellar interiors, escape without further interaction, and thus drain energy directly from the stellar interior. Hence, they contribute directly to the stellar energy losses, thus modifying stellar evolution. Our goal is to re-examine such constraints in the light of modern data and updated stellar evolution codes, paying particular attention to systematic and statistical errors. As a first example, we consider the case of a neutrino magnetic moment that enhances the energy loss from the plasma process. In terms of the observed color-magnitude diagrams, the tip of the red giant branch (RGB has been identified as a sensitive observable of the effects of the energy losses due to a neutrino magnetic moment. Here we describe the consequences of adding the cooling effect due to a neutrino magnetic moment to the Princeton-Goddard-PUC (PGPUC stellar evolution code, exploring in particular the dependence of the position of the RGB tip on the neutrino magnetic moment. As a first application, we studied the position of the observed RGB tip in the case of the Galactic globular cluster M5 (NGC 5904, using the latest, high-precision, ground-based data from the P. B. Stetson database (2012, priv. comm.. We compare the empirical results with the PGPUC model predictions, and discuss the implied constraints on the value of the neutrino magnetic moment.

  4. Enterotoxigenic profiles and polymerase chain reaction detection of Bacillus cereus group cells and B. cereus strains from foods and food-borne outbreaks.

    Science.gov (United States)

    Hsieh, Y M; Sheu, S J; Chen, Y L; Tsen, H Y

    1999-10-01

    Bacillus cereus is one of the important food pathogens. Since B. cereus group cells, such as B. cereus, B. thuringiensis, B. anthracis and B. mycoides, share many phenotypical properties and a high level of chromosomal sequence similarity, it is interesting to investigate the virulence profiles for B. cereus group cells, including B. cereus strains isolated from foods and samples associated with food-poisoning outbreaks. For this investigation, the presence of enterotoxin genes, such as those of haemolysin BL, B. cereus enterotoxin T and enterotoxin FM, were assayed by polymerase chain reaction (PCR) methods. Meanwhile, their enterotoxin activities were assayed using the BCET-RPLA kit, haemolytic patterns on sheep blood agar and their cytotoxicity to Chinese hamster ovary (CHO) cells. Results showed that there were 12 enterotoxigenic profiles for the 98 B. cereus group strains collected. In addition, if any of the three types of enterotoxins was present in the B. cereus group cells, these cells were shown to be cytotoxic to the CHO cells. Similar enterotoxigenic profiles could be found among strains of B. cereus, B. mycoides and B. thuringiensis. Thus, all B. cereus group strains may be potentially toxigenic and the detection of these cells in foods is important. We thus designed PCR primers, termed Ph1/Ph2, from the sphingomyelinase gene of B. cereus cells. These primers were specific for all B. cereus group strains and could be used for the detection of B. cereus cells contaminated in food samples.

  5. M/M/1/m系统算子的本征值特性(m=5,6)%The Characteristics of Eigenvalue of M/M/1/m(m =5,6)

    Institute of Scientific and Technical Information of China (English)

    盖平; 高超; 唐慧; 赵立杰

    2010-01-01

    研究了m=5,6时,M/M/1/m算子本征值特性:m=6时相应本征值的代数重为1;m=5,6时,相应的系统算子的非零本征值相互交替;m=6时的最大非零本征值比m=5时更靠近0点.这种特性延续了m=1,2,3,4,5时相应的特性.另外给出了m=5,6时,相应的po(t)图像.

  6. Do variations in mast cell hyperplasia account for differences in radiation-induced lung injury among different mouse strains, rats and nonhuman primates?

    Science.gov (United States)

    Down, Julian D; Medhora, Meetha; Jackson, Isabel L; Cline, J Mark; Vujaskovic, Zeljko

    2013-08-01

    The role of mast cell infiltrates in the pathology of radiation damage to the lung has been a subject of continuing investigation over the past four decades. This has been accompanied by a number of proposals as to how mast cells and the secretory products thereof participate in the generation of acute inflammation (pneumonitis) and the chronic process of collagen deposition (fibrosis). An additional pathophysiology examines the possible connection between mast cell hyperplasia and pulmonary hypertension through the release of vasoactive mediators. The timing and magnitude of pneumonitis and fibrosis are known to vary tremendously among different genetic mouse strains and animal species. Therefore, we have systematically compared mast cell numbers in lung sections from nine mouse strains, two rat strains and nonhuman primates (NHP) after whole thorax irradiation (WTI) at doses ranging from 10-15 Gy and at the time of entering respiratory distress. Mice of the BALB/c strain had a dramatic increase in interstitial mast cell numbers, similar to WAG/Rij and August rats, while relatively low levels of mast cell infiltrate were observed in other mouse strains (CBA, C3H, B6, C57L, WHT and TO mice). Enumeration of mast cell number in five NHPs (rhesus macaque), exhibiting severe pneumonitis at 17 weeks after 10 Gy WTI, also indicated a low response shared by the majority of mouse strains. There appeared to be no relationship between the mast cell response and the strain-dependent susceptibility towards pneumonitis or fibrosis. Further investigations are required to explore the possible participation of mast cells in mediating specific vascular responses and whether a genetically diverse mast cell response occurs in humans.

  7. Extracellular signal-regulated kinases 1/2 control claudin-2 expression in Madin-Darby canine kidney strain I and II cells.

    Science.gov (United States)

    Lipschutz, Joshua H; Li, Shixiong; Arisco, Amy; Balkovetz, Daniel F

    2005-02-01

    The tight junction of the epithelial cell determines the characteristics of paracellular permeability across epithelium. Recent work points toward the claudin family of tight junction proteins as leading candidates for the molecular components that regulate paracellular permeability properties in epithelial tissues. Madin-Darby canine kidney (MDCK) strain I and II cells are models for the study of tight junctions and based on transepithelial electrical resistance (TER) contain "tight" and "leaky" tight junctions, respectively. Overexpression studies suggest that tight junction leakiness in these two strains of MDCK cells is conferred by expression of the tight junction protein claudin-2. Extracellular signal-regulated kinase (ERK) 1/2 activation by hepatocyte growth factor treatment of MDCK strain II cells inhibited claudin-2 expression and transiently increased TER. This process was blocked by the ERK 1/2 inhibitor U0126. Transfection of constitutively active mitogen-activated protein kinase/extracellular signal-regulated kinase kinase into MDCK strain II cells also inhibited claudin-2 expression and increased TER. MDCK strain I cells have higher levels of active ERK 1/2 than do MDCK strain II cells. U0126 treatment of MDCK strain I cells decreased active ERK 1/2 levels, induced expression of claudin-2 protein, and decreased TER by approximately 20-fold. U0126 treatment also induced claudin-2 expression and decreased TER in a high resistance mouse cortical collecting duct cell line (94D). These data show for the first time that the ERK 1/2 signaling pathway negatively controls claudin-2 expression in mammalian renal epithelial cells and provide evidence for regulation of tight junction paracellular transport by alterations in claudin composition within tight junction complexes.

  8. Quantification of green fluorescent protein by in vivo imaging, PCR, and flow cytometry: comparison of transgenic strains and relevance for fetal cell microchimerism.

    Science.gov (United States)

    Fujiki, Yutaka; Tao, Kai; Bianchi, Diana W; Giel-Moloney, Maryann; Leiter, Andrew B; Johnson, Kirby L

    2008-02-01

    Animal models are increasingly being used for the assessment of fetal cell microchimerism in maternal tissue. We wished to determine the optimal transgenic mouse strain and analytic technique to facilitate the detection of rare transgenic microchimeric fetal cells amongst a large number of maternal wild-type cells. We evaluated two strains of mice transgenic for the enhanced green fluorescent protein (EGFP): a commercially available, commonly used strain (C57BL/6-Tg(ACTB-EGFP)10sb/J) (CAG) and a newly created strain (ROSA26-EGFP) using three different techniques: in vivo and ex vivo fluorescent imaging (for whole body and dissected organs, respectively), PCR amplification of gfp, and flow cytometry (FCM). By fluorescent imaging, organs from CAG mice were 10-fold brighter than organs from ROSA26-EGFP mice (P PCR, more transgene from CAG mice was detected compared to ROSA26-EGFP mice (P = 0.04). By FCM, ROSA26-EGFP cell fluorescence was more uniform than CAG cells. A greater proportion of cells from ROSA26-EGFP organs were positive for EGFP than cells from CAG organs, but CAG mice had a greater proportion of cells with the brightest fluorescent intensity. Each transgenic strain possesses characteristics that make it useful under specific experimental circumstances. The CAG mouse model is preferable when experiments require brighter cells, whereas ROSA26-EGFP is more appropriate when uniform or ubiquitous expression is more important than brightness. Investigators must carefully select the transgenic strain most suited to the experimental design to obtain the most consistent and reproducible data. In vivo imaging allows for phenotypic evaluation of whole animals and intact organs; however, we did not evaluate its utility for the detection of rare, fetal microchimeric cells in the maternal organs. Finally, while PCR amplification of a paternally inherited transgene does allow for the quantitative determination of rare microchimeric cells, FCM allows for both

  9. Antifungal activity against Cryptococcus neoformans strains and genotoxicity assessment in human leukocyte cells of Euphorbia tirucalli L.

    Directory of Open Access Journals (Sweden)

    Luís Flávio Souza de Oliveira

    2014-12-01

    Full Text Available In the last times, focus on plant research has increased all over the world. Euphorbia tirucalli L., a plant known popularly as Aveloz, and originally used in Africa, has been drawing attention for its use in the United States and Latin America, both for use as an ornamental plant and as a medicinal plant. E. tirucalli L. is a member of the family Euphorbiaceae and contains many diterpenoids and triterpenoids, in particular phorbol esters, apparently the main constituent of this plant, which are assumed to be responsible for their activities in vivo and in vitro. The in vitro antifungal activities of Euphorbia tirucalli (L. against opportunistic yeasts were studied using microbroth dilution assay. The results showed that aqueous extract and latex preparation were effective against ten clinical strains of Cryptococcus neoformans in vitro (Latex and extract MIC range of 3.2 - > 411 µg/mL. Aiming the safe use in humans, the genotoxic effects of E. tirucalli were evaluated in human leukocytes cells. Our data show that both aqueous extract and latex preparation have no genotoxic effect in human leukocytes cells in vitro. Although the results cannot be extrapolated by itself for use in vivo, they suggest a good perspective for a therapeutic application in future. In conclusion, our results show that the aqueous extract and latex preparation from E. tirucalli L. are antifungal agents effectives against several strains of C. neoformans and do not provoke DNA damage in human leukocyte cells, considering the concentrations tested.

  10. Carbohydrate-binding specificities of potential probiotic Lactobacillus strains in porcine jejunal (IPEC-J2) cells and porcine mucin.

    Science.gov (United States)

    Valeriano, Valerie Diane; Bagon, Bernadette B; Balolong, Marilen P; Kang, Dae-Kyung

    2016-07-01

    Bacterial lectins are carbohydrate-binding adhesins that recognize glycoreceptors in the gut mucus and epithelium of hosts. In this study, the contribution of lectin-like activities to adhesion of Lactobacillus mucosae LM1 and Lactobacillus johnsonii PF01, which were isolated from swine intestine, were compared to those of the commercial probiotic Lactobacillus rhamnosus GG. Both LM1 and PF01 strains have been reported to have good adhesion ability to crude intestinal mucus of pigs. To confirm this, we quantified their adhesion to porcine gastric mucin and intestinal porcine enterocytes isolated from the jejunum of piglets (IPEC-J2). In addition, we examined their carbohydrate-binding specificities by suspending bacterial cells in carbohydrate solutions prior to adhesion assays. We found that the selected carbohydrates affected the adherences of LM1 to IPEC-J2 cells and of LGG to mucin. In addition, compared to adhesion to IPEC-J2 cells, adhesion to mucin by both LM1 and LGG was characterized by enhanced specific recognition of glycoreceptor components such as galactose, mannose, and N-acetylglucosamine. Hydrophobic interactions might make a greater contribution to adhesion of PF01. A similar adhesin profile between a probiotic and a pathogen, suggest a correlation between shared pathogen-probiotic glycoreceptor recognition and the ability to exclude enteropathogens such as Escherichia coli K88 and Salmonella Typhimurium KCCM 40253. These findings extend our understanding of the mechanisms of the intestinal adhesion and pathogen-inhibition abilities of probiotic Lactobacillus strains.

  11. Damage and Shaking Intensity in the M5.7 Canyondam Earthquake

    Science.gov (United States)

    Boatwright, J.; Chapman, K.; Gold, M. B.; Hardebeck, J. L.

    2013-12-01

    An M5.7 earthquake occurred southeast of Lake Almanor, CA, at 8:47 PM on May 23, 2013. Double-difference relocations of the main shock and aftershocks indicate that the earthquake nucleated at 11 km depth and ruptured up dip on a fault striking 292° and dipping 70° to the northeast. The earthquake cracked foundations, broke chimneys, and ruptured plumbing around Lake Almanor. We canvassed communities around the lake and to the south and east for earthquake damage, adding reports from our interviews to the geocoded 'Did You Feel It?' reports and to a set of damage reports collected by the Plumas County Office of Emergency Services. Three communities suffered significant damage. In Lake Almanor West, 14 km and 290° from the hypocenter, one wood-frame house was shifted on its foundation, the cripple wall of another house was racked, and water and gas pipes in five houses were ruptured. This damage indicates the shaking approached MMI 8. In Lake Almanor Country Club, 10 km and 310° from the hypocenter, more than 40 chimneys were cracked, broken, or collapsed, a coupling for the municipal water tank was ruptured, and a 200-foot long fissure opened on a slope facing the lake. This damage indicates shaking between MMI 7 and MMI 8, consistent with the accelerograph recording of PGA = 38% g and PGV = 30 cm/s at the Fire Station in Lake Almanor Country Club. This CSMIP station and a PG&E station on the crest of the Butt Valley Dam obtained the only recordings within 50 km of the epicenter. In Hamilton Branch, 10 km and 345° from the hypocenter, a foundation of a wood-frame house was damaged, and 14 chimneys and a water pipe were broken, indicative of MMI 7 shaking. All three communities are underlain by Tertiary and Quaternary basalts. The communities of Chester, Westwood, and Greenville were less damaged, suffering cracked drywall, broken windows, and objects thrown from shelves. The intensities in the three most strongly damaged communities increase as the azimuth

  12. Structure and heterologous expression of the gene encoding the cell surface glycoprotein from Haloarcula japonica strain TR-1.

    Science.gov (United States)

    Wakai, H; Takada, K; Nakamura, S; Horikoshi, K

    1995-01-01

    The gene encoding the cell surface glycoprotein (CSG) of Haloarcula japonica strain TR-1 was cloned and sequenced. The structural gene consisted from an open reading frame of 2,586 bp. A potential promoter sequence was found about 150 bp upstream of the ATG initiation codon. N-terminal amino acid sequence of the Ha. japonica CSG revealed that the mature CSG consisted of 828 amino acids. Five potential N-glycosylation sites were found in the mature sequence. The cloned CSG gene of Ha. japonica was expressed in closely-related halophilic archaea.

  13. A combined M5P tree and hazard-based duration model for predicting urban freeway traffic accident durations.

    Science.gov (United States)

    Lin, Lei; Wang, Qian; Sadek, Adel W

    2016-06-01

    The duration of freeway traffic accidents duration is an important factor, which affects traffic congestion, environmental pollution, and secondary accidents. Among previous studies, the M5P algorithm has been shown to be an effective tool for predicting incident duration. M5P builds a tree-based model, like the traditional classification and regression tree (CART) method, but with multiple linear regression models as its leaves. The problem with M5P for accident duration prediction, however, is that whereas linear regression assumes that the conditional distribution of accident durations is normally distributed, the distribution for a "time-to-an-event" is almost certainly nonsymmetrical. A hazard-based duration model (HBDM) is a better choice for this kind of a "time-to-event" modeling scenario, and given this, HBDMs have been previously applied to analyze and predict traffic accidents duration. Previous research, however, has not yet applied HBDMs for accident duration prediction, in association with clustering or classification of the dataset to minimize data heterogeneity. The current paper proposes a novel approach for accident duration prediction, which improves on the original M5P tree algorithm through the construction of a M5P-HBDM model, in which the leaves of the M5P tree model are HBDMs instead of linear regression models. Such a model offers the advantage of minimizing data heterogeneity through dataset classification, and avoids the need for the incorrect assumption of normality for traffic accident durations. The proposed model was then tested on two freeway accident datasets. For each dataset, the first 500 records were used to train the following three models: (1) an M5P tree; (2) a HBDM; and (3) the proposed M5P-HBDM, and the remainder of data were used for testing. The results show that the proposed M5P-HBDM managed to identify more significant and meaningful variables than either M5P or HBDMs. Moreover, the M5P-HBDM had the lowest overall mean

  14. Strain Stiffening of Fibrillar Collagen during Individual and Collective Cell Migration Identified by AFM Nanoindentation

    NARCIS (Netherlands)

    Helvert, S. van; Friedl, P.

    2016-01-01

    The multistep process of cell migration requires cells to dynamically couple to extracellular interfaces and generate traction force or friction for displacement of the cell body. When deformed, biopolymer networks, including fibrillar collagen and fibrin, undergo a nonlinear elasticity change that

  15. YccW is the m5C methyltransferase specific for 23S rRNA nucleotide 1962

    DEFF Research Database (Denmark)

    Purta, Elzbieta; O'Connor, Michelle; Bujnicki, Janusz M

    2008-01-01

    . coli marginally reduces its growth rate. YccW had previously eluded identification because it displays only limited sequence similarity to the m(5)C methyltransferases RsmB and RsmF and is in fact more similar to known m(5)U (5-methyluridine) RNA methyltransferases. In keeping with the previously...... proposed nomenclature system for bacterial rRNA methyltransferases, yccW is now designated as the rRNA large subunit methyltransferase gene rlmI....

  16. Dysregulated Circulating Dendritic Cell Function in Ulcerative Colitis Is Partially Restored by Probiotic Strain Lactobacillus casei Shirota

    Directory of Open Access Journals (Sweden)

    Elizabeth R. Mann

    2013-01-01

    Full Text Available Background. Dendritic cells regulate immune responses to microbial products and play a key role in ulcerative colitis (UC pathology. We determined the immunomodulatory effects of probiotic strain Lactobacillus casei Shirota (LcS on human DC from healthy controls and active UC patients. Methods. Human blood DC from healthy controls (control-DC and UC patients (UC-DC were conditioned with heat-killed LcS and used to stimulate allogeneic T cells in a 5-day mixed leucocyte reaction. Results. UC-DC displayed a reduced stimulatory capacity for T cells (P<0.05 and enhanced expression of skin-homing markers CLA and CCR4 on stimulated T cells (P<0.05 that were negative for gut-homing marker β7. LcS treatment restored the stimulatory capacity of UC-DC, reflecting that of control-DC. LcS treatment conditioned control-DC to induce CLA on T cells in conjunction with β7, generating a multihoming profile, but had no effects on UC-DC. Finally, LcS treatment enhanced DC ability to induce TGFβ production by T cells in controls but not UC patients. Conclusions. We demonstrate a systemic, dysregulated DC function in UC that may account for the propensity of UC patients to develop cutaneous manifestations. LcS has multifunctional immunoregulatory activities depending on the inflammatory state; therapeutic effects reported in UC may be due to promotion of homeostasis.

  17. Sialidases affect the host cell adherence and epsilon toxin-induced cytotoxicity of Clostridium perfringens type D strain CN3718.

    Directory of Open Access Journals (Sweden)

    Jihong Li

    2011-12-01

    Full Text Available Clostridium perfringens type B or D isolates, which cause enterotoxemias or enteritis in livestock, produce epsilon toxin (ETX. ETX is exceptionally potent, earning it a listing as a CDC class B select toxin. Most C. perfringens strains also express up to three different sialidases, although the possible contributions of those enzymes to type B or D pathogenesis remain unclear. Type D isolate CN3718 was found to carry two genes (nanI and nanJ encoding secreted sialidases and one gene (nanH encoding a cytoplasmic sialidase. Construction in CN3718 of single nanI, nanJ and nanH null mutants, as well as a nanI/nanJ double null mutant and a triple sialidase null mutant, identified NanI as the major secreted sialidase of this strain. Pretreating MDCK cells with NanI sialidase, or with culture supernatants of BMC206 (an isogenic CN3718 etx null mutant that still produces sialidases enhanced the subsequent binding and cytotoxic effects of purified ETX. Complementation of BMC207 (an etx/nanH/nanI/nanJ null mutant showed this effect is mainly attributable to NanI production. Contact between BMC206 and certain mammalian cells (e.g., enterocyte-like Caco-2 cells resulted in more rapid sialidase production and this effect involved increased transcription of BMC206 nanI gene. BMC206 was shown to adhere to some (e.g. Caco-2 cells, but not all mammalian cells, and this effect was dependent upon sialidase, particularly NanI, expression. Finally, the sialidase activity of NanI (but not NanJ or NanH could be enhanced by trypsin. Collectively these in vitro findings suggest that, during type D disease originating in the intestines, trypsin may activate NanI, which (in turn could contribute to intestinal colonization by C. perfringens type D isolates and also increase ETX action.

  18. Ethanol production by a flocculant yeast strain in a CSTR type fermentor with cell recycling.

    Science.gov (United States)

    Hojo, O; Hokka, C O; Major, A M

    1999-01-01

    Tests were performed in a continuous stirred tank reactor (CSTR), with and without cell recycling, to produce ethanol. The reactor without cell recycling produced the kinetic model of ethanol production, whereas the reactor with cell recycling allowed for a study of process stability. The Levenspiel kinetic model was adopted; however, in the case of fermentation with cell recycling, the coefficient of cell death was added. It was observed that cellular viability varied greatly throughout the fermenting process and that microaeration is of fundamental importance in maintaining the stability of the process.

  19. Effect of diffusely adherent Escherichia coli strains isolated from diarrhoeal patients and healthy carriers on IL-8 secretion and tight junction barrier integrity of Caco-2 cells.

    Science.gov (United States)

    Tanimoto, Yoshihiko; Arikawa, Kentaro; Nishikawa, Yoshikazu

    2013-03-15

    The pathogenesis of diffusely adherent Escherichia coli (DAEC) remains to be elucidated. Previously, we found that afimbrial adhesin gene (afa)-positive motile DAEC strains isolated from patients with diarrhoea induce high levels of IL-8 secretion in Caco-2 cells via toll-like receptor 5 (TLR-5), while non-motile strains did not. The aim of this study was to compare virulence properties, including the phylogenetic groups, afa subtypes, IL-8 secretion levels, and the effects on tight junctions, of DAEC strains isolated from healthy persons with those isolated from patients with diarrhoea. Induction of IL-8 secretion in Caco-2 cells was examined for a total of 36 afa-positive strains: 19 from diarrhoeal patients and 17 from healthy carriers. Irrespective of the source, all strains were classified into the phylogenetic group B2 or D, with the exception of two strains. All 7 motile strains isolated from diarrhoeal patients induced high levels of IL-8 secretion, while only 6 of 15 motile strains from healthy carriers induced IL-8 secretion to the same levels as the diarrhoeal strains. We speculated that additional virulence factors other than afa and motility cause the loosening of tight junctions that allows flagellin to reach TLR-5 located on the basolateral side of the epithelium. However, no differences in the TER and dextran permeability were observed between cells infected with diarrhoeal strains and those from healthy persons. Thus, diarrhoeagenic DAEC seems to possess additional factors, in addition to adhesin and flagellin, which can induce high IL-8 secretion.

  20. Dendritic Cell Response to HIV-1 Is Controlled by Differentiation Programs in the Cells and Strain-Specific Properties of the Virus.

    Science.gov (United States)

    Nasi, Aikaterini; Amu, Sylvie; Göthlin, Mårten; Jansson, Marianne; Nagy, Noemi; Chiodi, Francesca; Réthi, Bence

    2017-01-01

    Dendritic cells (DCs) are potent antigen-presenting cells that might play contradictory roles during HIV-1 infection, contributing not only to antiviral immunity but also to viral dissemination and immune evasion. Although DCs are characterized by enormous functional diversity, it has not been analyzed how differentially programmed DCs interact with HIV-1. We have previously described the reprogramming of DC development by endogenously produced lactic acid that accumulated in a cell culture density-dependent manner and provided a long-lasting anti-inflammatory signal to the cells. By exploiting this mechanism, we generated immunostimulatory DCs characterized by the production of TH1 polarizing and inflammatory mediators or, alternatively, suppressed DCs that produce IL-10 upon activation, and we tested the interaction of these DC types with different HIV-1 strains. Cytokine patterns were monitored in HIV-1-exposed DC cultures. Our results showed that DCs receiving suppressive developmental program strongly upregulated their capacity to produce the TH1 polarizing cytokine IL-12 and the inflammatory chemokines CCL2 and CCL7 upon interaction with HIV-1 strains IIIB and SF162. On the contrary, HIV-1 abolished cytokine production in the more inflammatory DC types. Preincubation of the cells with the HIV-1 proteins gp120 and Nef could inhibit IL-12 production irrespectively of the tested DC types, whereas MyD88- and TRIF-dependent signals stimulated IL-12 production in the suppressed DC type only. Rewiring of DC cytokines did not require DC infections or ligation of the HIV-1 receptor CD209. A third HIV-1 strain, BaL, could not modulate DC cytokines in a similar manner indicating that individual HIV-1 strains can differ in their capacity to influence DCs. Our results demonstrated that HIV-1 could not induce definite and invariable modulatory programs in DCs. Instead, interaction with the virus triggered different responses in different DC types. Thus, the outcome of DC

  1. Dendritic Cell Response to HIV-1 Is Controlled by Differentiation Programs in the Cells and Strain-Specific Properties of the Virus

    Science.gov (United States)

    Nasi, Aikaterini; Amu, Sylvie; Göthlin, Mårten; Jansson, Marianne; Nagy, Noemi; Chiodi, Francesca; Réthi, Bence

    2017-01-01

    Dendritic cells (DCs) are potent antigen-presenting cells that might play contradictory roles during HIV-1 infection, contributing not only to antiviral immunity but also to viral dissemination and immune evasion. Although DCs are characterized by enormous functional diversity, it has not been analyzed how differentially programmed DCs interact with HIV-1. We have previously described the reprogramming of DC development by endogenously produced lactic acid that accumulated in a cell culture density-dependent manner and provided a long-lasting anti-inflammatory signal to the cells. By exploiting this mechanism, we generated immunostimulatory DCs characterized by the production of TH1 polarizing and inflammatory mediators or, alternatively, suppressed DCs that produce IL-10 upon activation, and we tested the interaction of these DC types with different HIV-1 strains. Cytokine patterns were monitored in HIV-1-exposed DC cultures. Our results showed that DCs receiving suppressive developmental program strongly upregulated their capacity to produce the TH1 polarizing cytokine IL-12 and the inflammatory chemokines CCL2 and CCL7 upon interaction with HIV-1 strains IIIB and SF162. On the contrary, HIV-1 abolished cytokine production in the more inflammatory DC types. Preincubation of the cells with the HIV-1 proteins gp120 and Nef could inhibit IL-12 production irrespectively of the tested DC types, whereas MyD88- and TRIF-dependent signals stimulated IL-12 production in the suppressed DC type only. Rewiring of DC cytokines did not require DC infections or ligation of the HIV-1 receptor CD209. A third HIV-1 strain, BaL, could not modulate DC cytokines in a similar manner indicating that individual HIV-1 strains can differ in their capacity to influence DCs. Our results demonstrated that HIV-1 could not induce definite and invariable modulatory programs in DCs. Instead, interaction with the virus triggered different responses in different DC types. Thus, the outcome of DC

  2. Role of interfacial strain in fiber-shaped solar cell based on TiO2 nanotube arrays.

    Science.gov (United States)

    Fan, Xing; Huang, Lu; Liu, Zuohua; Tao, Changyuan

    2014-09-01

    This study reports the first equivalent circuit model for all-solid, fiber-shaped, dye-sensitized solar cell (DSSC), in order to reveal the internal catalytic reaction mechanism in this new type of solar cells. The counter electrode of the winding structure leads to negative impedance under high frequency, which is consistent with the model. The study further investigates the strain of the TiO2 nanotube (TNT) arrays and its influence on interfacial mechanism. As a unique characteristic of fiber-shaped DSSC, the strain of the TNT arrays strengthens the permeation of the electrolyte. The permeation not only improves the efficiency of interfacial photochemical reactions, but also magnifies the probability of the side reactions on the electrolyte/Ti interfaces. Therefore, both the variation of impedance and overall conversion efficiency exhibit similar inflection points. Different from that of traditional plate-type device, the interfacial impedance in the equivalent circuit of fiber-shaped devices should be treated as a variable for changes in TiO2 and CuI layers.

  3. Biotransformation of indole to indigo by the whole cells of phenol hydroxylase engineered strain in biphasic systems.

    Science.gov (United States)

    Shi, Shengnan; Ma, Fang; Sun, Tieheng; Li, Ang; Zhou, Jiti; Qu, Yuanyuan

    2013-02-01

    Biotransformation of indole to indigo in liquid-liquid biphasic systems was performed in Escherichia coli cells expressing phenol hydroxylase. It was suggested that indole could inhibit the cell growth even at low concentration of 0.1 g/L. The critical Log P for strain PH_(IND) was about 5.0. Three different solvents, i.e., decane, dodecane, and dioctyl phthalate, were selected as organic phase in biphasic media. The results showed that dodecane gave the highest yield of indigo (176.4 mg/L), which was more than that of single phase (90.5 mg/L). The optimal conditions for biotransformation evaluated by response surface methodology were as follows: 540.26 mg/L of indole concentration, 42.27 % of organic phase ratio, and 200 r/min of stirrer speed; under these conditions, the maximal production of indigo was 243.51 mg/L. This study proved that the potential application of strain PH_(IND) in the biotransformation of indole to indigo using liquid-liquid biphasic systems.

  4. RNA of Enterococcus faecalis Strain EC-12 Is a Major Component Inducing Interleukin-12 Production from Human Monocytic Cells.

    Directory of Open Access Journals (Sweden)

    Ryoichiro Nishibayashi

    Full Text Available Interleukin-12 (IL-12 is an important cytokine for the immunomodulatory effects of lactic acid bacteria (LAB. Using murine immune cells, we previously reported that the RNA of Enterococcus faecalis EC-12, a LAB strain exerting probiotic-like beneficial effects, is the major IL-12-inducing immunogenic component. However, it was recently revealed that bacterial RNA can be a ligand for Toll-like receptor (TLR 13, which is only expressed in mice. Because TLR13 is not expressed in humans, the immuno-stimulatory and -modulatory effects of LAB RNA in human cells should be augmented excluding TLR13 contribution. In experiment 1 of this study, the role of LAB RNA in IL-12 induction in human immune cells was studied using three LAB strains, E.faecalis EC-12, Lactobacillus gasseri JCM5344, and Bifidobacterium breve JCM1192. RNase A treatment of heat-killed LAB significantly decreased the IL-12 production of human peripheral blood mononuclear cells on stimulation, while RNase III treatment revealed virtually no effects. Further, IL-12 production against heat-killed E. faecalis EC-12 was abolished by depleting monocytes. These results demonstrated that single stranded RNA (ssRNA of LAB is a strong inducer of IL-12 production from human monocytes. In experiment 2, major receptor for ssRNA of E. faecalis EC-12 was identified using THP-1 cells, a human monocytic cell line. The type of RNA molecules of E. faecalis EC-12 responsible for IL-12 induction was also identified. IL-12 production induced by the total RNA of E. faecalis EC-12 was significantly reduced by the treatment of siRNA for TLR8 but not for TLR7. Furthermore, both 23S and 16S rRNA, but not mRNA, of E. faecalis EC-12 markedly induced IL-12 production from THP-1 cells. These results suggested that the recognition of ssRNA of E. faecalis EC-12 was mediated by TLR8 and that rRNA was the RNA molecule that exhibited IL-12-inducing ability in human cells.

  5. CD8+ T-Cell responses to Trypanosoma cruzi are highly focused on strain-variant trans-sialidase epitopes.

    Directory of Open Access Journals (Sweden)

    Diana L Martin

    2006-08-01

    Full Text Available CD8+ T cells are crucial for control of a number of medically important protozoan parasites, including Trypanosoma cruzi, the agent of human Chagas disease. Yet, in contrast to the wealth of information from viral and bacterial infections, little is known about the antigen specificity or the general development of effector and memory T-cell responses in hosts infected with protozoans. In this study we report on a wide-scale screen for the dominant parasite peptides recognized by CD8+ T cells in T. cruzi-infected mice and humans. This analysis demonstrates that in both hosts the CD8+ T-cell response is highly focused on epitopes encoded by members of the large trans-sialidase family of genes. Responses to a restricted set of immunodominant peptides were especially pronounced in T. cruzi-infected mice, with more than 30% of the CD8+ T-cell response at the peak of infection specific for two major groups of trans-sialidase peptides. Experimental models also demonstrated that the dominance patterns vary depending on the infective strain of T. cruzi, suggesting that immune evasion may be occurring at a population rather than single-parasite level.

  6. Engineering fibrin-based tissue constructs from myofibroblasts and application of constraints and strain to induce cell and collagen reorganization.

    Science.gov (United States)

    de Jonge, Nicky; Baaijens, Frank P T; Bouten, Carlijn V C

    2013-10-28

    Collagen content and organization in developing collagenous tissues can be influenced by local tissue strains and tissue constraint. Tissue engineers aim to use these principles to create tissues with predefined collagen architectures. A full understanding of the exact underlying processes of collagen remodeling to control the final tissue architecture, however, is lacking. In particular, little is known about the (re)orientation of collagen fibers in response to changes in tissue mechanical loading conditions. We developed an in vitro model system, consisting of biaxially-constrained myofibroblast-seeded fibrin constructs, to further elucidate collagen (re)orientation in response to i) reverting biaxial to uniaxial static loading conditions and ii) cyclic uniaxial loading of the biaxially-constrained constructs before and after a change in loading direction, with use of the Flexcell FX4000T loading device. Time-lapse confocal imaging is used to visualize collagen (re)orientation in a nondestructive manner. Cell and collagen organization in the constructs can be visualized in real-time, and an internal reference system allows us to relocate cells and collagen structures for time-lapse analysis. Various aspects of the model system can be adjusted, like cell source or use of healthy and diseased cells. Additives can be used to further elucidate mechanisms underlying collagen remodeling, by for example adding MMPs or blocking integrins. Shape and size of the construct can be easily adapted to specific needs, resulting in a highly tunable model system to study cell and collagen (re)organization.

  7. A Genotypic Analysis of Five P. aeruginosa Strains after Biofilm Infection by Phages Targeting Different Cell Surface Receptors

    Directory of Open Access Journals (Sweden)

    Diana P. Pires

    2017-06-01

    Full Text Available Antibiotic resistance constitutes one of the most serious threats to the global public health and urgently requires new and effective solutions. Bacteriophages are bacterial viruses increasingly recognized as being good alternatives to traditional antibiotic therapies. In this study, the efficacy of phages, targeting different cell receptors, against Pseudomonas aeruginosa PAO1 biofilm and planktonic cell cultures was evaluated over the course of 48 h. Although significant reductions in the number of viable cells were achieved for both cases, the high level of adaptability of the bacteria in response to the selective pressure caused by phage treatment resulted in the emergence of phage-resistant variants. To further investigate the genetic makeup of phage-resistant variants isolated from biofilm infection experiments, some of these bacteria were selected for phenotypic and genotypic characterization. Whole genome sequencing was performed on five phage-resistant variants and all of them carried mutations affecting the galU gene as well as one of pil genes. The sequencing analysis further revealed that three of the P. aeruginosa PAO1 variants carry large deletions (>200 kbp in their genomes. Complementation of the galU mutants with wild-type galU in trans restored LPS expression on the bacterial cell surface of these bacterial strains and rendered the complemented strains to be sensitive to phages. This provides unequivocal evidence that inactivation of galU function was associated with resistance to the phages that uses LPS as primary receptors. Overall, this work demonstrates that P. aeruginosa biofilms can survive phage attack and develop phage-resistant variants exhibiting defective LPS production and loss of type IV pili that are well adapted to the biofilm mode of growth.

  8. Comparison of Artificial Neural Network And M5 Model Tree Technique In Water Level Forecasting of Solo River

    Science.gov (United States)

    Lasminto, Umboro; Hery Mularta, Listya

    2010-05-01

    Flood events along the Solo River flow at the end of December 2007 has caused lose of properties and lives. Floods occurred in the city of Ngawi, Madiun, Bojonegoro, Babat and surrounding areas. To reduce future losses, one of the important efforts that will occur during a flood is to get information about the magnitude and time will be floods, so that people can make an effort to reduce its impact. Flood forecasting model can provide information of water level in the river some time before the incident. This paper will compare the flood forecasting model at Bojonegoro City was built using the technique of Artificial Neural Network (ANN) and M5 Model Tree (M5MT). The model will forecast the water level of 1, 3 and 6 hours ahead at the point of water level recorders in the City of Bojonegoro using input from the water level at some point water level recorders in the upstream such as Karangnongko, Sekayu, Jurug and Wonogiri. The same data set of hourly water level records are used to build the model of ANN and M5MT technique. The selection of parameters and setup of ANN and M5MT technique is done to obtain the best result. The results of the model are evaluated by calculating the Root Mean Square Error (RMSE) between the predictions and observations. RMSE produced by the water level forecasting model 1, 3 and 6 hours ahead with M5MT technique are 0.2723, 0.6279 and 0.7176 meters. While the ANN technique are 0.1829, 0.3192 and 0517 meters. ANN technique has a better ability in predicting low flow, whereas M5 Model Tree technique has a better ability in predicting high flow. Keywords : Water level forecasting, Solo River, M5 Model Tree, Artificial Neural Network

  9. Effects of the strain background and autolysis process on the composition and biophysical properties of the cell wall from two different industrial yeasts.

    Science.gov (United States)

    Schiavone, Marion; Sieczkowski, Nathalie; Castex, Mathieu; Dague, Etienne; Marie François, Jean

    2015-03-01

    The Saccharomyces cerevisiae cell surface is endowed with some relevant technological properties, notably antimicrobial and biosorption activities. For these purposes, yeasts are usually processed and packaged in an 'autolysed/dried' formula, which may have some impacts on cell surface properties. In this report, we showed using a combination of biochemical, biophysical and molecular methods that the composition of the cell wall of two wine yeast strains was not altered by the autolysis process. In contrast, this process altered the nanomechanical properties as shown by a 2- to 4-fold increased surface roughness and to a higher adhesion to the atomic force microscope tips of the autolysed cells as compared to live yeast cells. Besides, we found that the two strains harboured differences in biomechanical properties that could be due in part to higher levels of mannan in one of them, and to the fact that the surface of this mannan-enriched strain is decorated with highly adhesive patches forming nanodomains. The presence of these nanodomains could be correlated with the upregulation of flocculin encoding FLO11 as well as to higher expression of few other genes encoding cell wall mannoproteins in this mannan-enriched strain as compared to the other strain.

  10. Adhesion, invasion, intracellular survival and cytotoxic activity of strains of Aeromonas spp. in HEp-2, Caco-2 and T-84 cell lines.

    Science.gov (United States)

    dos Santos, Paula Azevedo; Pereira, Ana Claudia Machado; Ferreira, Andréa Fonseca; de Mattos Alves, Maria Angélica; Rosa, Ana Cláudia Paula; Freitas-Almeida, Angela Corrêa

    2015-05-01

    The genus Aeromonas contains important pathogen for both humans and other animals, being responsible for the etiology of intestinal and extraintestinal diseases. The pathology caused by these bacteria involves several virulence factors, such as the ability to produce toxins, adhesion and invasion. The properties conferred by these factors have been extensively studied in experiments of interaction between bacterial strains and cell culture. We evaluate the interaction of eight Aeromonas spp. strains, previously isolated from human faeces, food and water with HEp-2, Caco-2 and T-84 cell lines. Cytotoxic effects, the pattern of adhesion, invasive capacity and intracellular survival were analyzed. The results showed that Aeromonas strains were adherent to three cells lines in 6 h of incubation, displaying the aggregative adherence pattern. Among eight strains studied, 50% produced cytotoxic effects on HEp-2 cells, while none of the strains produced cytotoxic effects on Caco-2 and T-84 cells at 48 h. This study demonstrated that subsets of Aeromonas isolated from different sources were able to invade intestinal (T-84, Caco-2) and epithelial (HEp-2) cell lines cultivated in vitro surviving in intracellular environments up to 72 h. Finally, our results support the pathogenic potential of Aeromonas, especially those of food and clinical sources.

  11. Tunable Expression Tools Enable Single-Cell Strain Distinction in the Gut Microbiome.

    Science.gov (United States)

    Whitaker, Weston R; Shepherd, Elizabeth Stanley; Sonnenburg, Justin L

    2017-04-20

    Applying synthetic biology to engineer gut-resident microbes provides new avenues to investigate microbe-host interactions, perform diagnostics, and deliver therapeutics. Here, we describe a platform for engineering Bacteroides, the most abundant genus in the Western microbiota, which includes a process for high-throughput strain modification. We have identified a novel phage promoter and translational tuning strategy and achieved an unprecedented level of expression that enables imaging of fluorescent-protein-expressing Bacteroides stably colonizing the mouse gut. A detailed characterization of the phage promoter has provided a set of constitutive promoters that span over four logs of strength without detectable fitness burden within the gut over 14 days. These promoters function predictably over a 1,000,000-fold expression range in phylogenetically diverse Bacteroides species. With these promoters, unique fluorescent signatures were encoded to allow differentiation of six species within the gut. Fluorescent protein-based differentiation of isogenic strains revealed that priority of gut colonization determines colonic crypt occupancy. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. The fraction of cells that resume growth after acetic acid addition is a strain-dependent parameter of acetic acid tolerance in Saccharomyces cerevisiae.

    Science.gov (United States)

    Swinnen, Steve; Fernández-Niño, Miguel; González-Ramos, Daniel; van Maris, Antonius J A; Nevoigt, Elke

    2014-06-01

    High acetic acid tolerance of Saccharomyces cerevisiae is a relevant phenotype in industrial biotechnology when using lignocellulosic hydrolysates as feedstock. A screening of 38 S. cerevisiae strains for tolerance to acetic acid revealed considerable differences, particularly with regard to the duration of the latency phase. To understand how this phenotype is quantitatively manifested, four strains exhibiting significant differences were studied in more detail. Our data show that the duration of the latency phase is primarily determined by the fraction of cells within the population that resume growth. Only this fraction contributed to the exponential growth observed after the latency phase, while all other cells persisted in a viable but non-proliferating state. A remarkable variation in the size of the fraction was observed among the tested strains differing by several orders of magnitude. In fact, only 11 out of 10(7)  cells of the industrial bioethanol production strain Ethanol Red resumed growth after exposure to 157 mM acetic acid at pH 4.5, while this fraction was 3.6 × 10(6) (out of 10(7)  cells) in the highly acetic acid tolerant isolate ATCC 96581. These strain-specific differences are genetically determined and represent a valuable starting point to identify genetic targets for future strain improvement.

  13. Identification, recombinant expression, and characterization of the 100 kDa high molecular weight Hymenoptera venom allergens Api m 5 and Ves v 3.

    Science.gov (United States)

    Blank, Simon; Seismann, Henning; Bockisch, Benjamin; Braren, Ingke; Cifuentes, Liliana; McIntyre, Mareike; Rühl, Dana; Ring, Johannes; Bredehorst, Reinhard; Ollert, Markus W; Grunwald, Thomas; Spillner, Edzard

    2010-05-01

    Insect stings can cause life-threatening IgE-mediated anaphylactic reactions in venom-allergic patients. Although several compounds have already been described as venom allergens, prominent allergen candidates especially in the higher m.w. range have still remained elusive. Tandem mass spectrometry-based sequencing assigned a candidate gene to the most prominent putative high m.w. allergen Api m 5 (allergen C) in honeybee (Apis mellifera) venom and also allowed identification of its homologue Ves v 3 in yellow jacket (Vespula vulgaris) venom. Both proteins exhibit a pronounced sequence identity to human dipeptidyl peptidase IV or CD26. Reactivity of a human IgE mAb verified the presence of these proteins in the venoms. Both proteins were produced in insect cells and characterized for their enzymatic activity as well as their allergenic potential using sera and basophils from insect venom-allergic patients. Both Api m 5 and Ves v 3 were recognized by specific IgE of the majority of patients even in the absence of cross-reactive carbohydrate determinants. Serologic IgE reactivity closely matched activation of human basophils by Api m 5 or Ves v 3, thus underlining their relevance in functional assays. With Api m 5 and Ves v 3, a new pair of homologous allergens becomes available for future clinical applications in diagnosis and therapy that may also contribute to the understanding of molecular mechanisms of insect venoms. Moreover, the patient IgE reactivity together with the cellular activation demonstrates for the first time the relevance of high m.w. allergens in the context of hymenoptera venom allergy.

  14. Relative susceptibility of six continuous cell lines for cultivation of chlamydia trachomatis strains

    Directory of Open Access Journals (Sweden)

    Malathi J

    2004-01-01

    Full Text Available Since susceptibility of a cell line is an important factor for cultivation of Chlamydia trachomatis, McCoy, HeLa, BHK-21, HEp-2, Vero and A549 cell lines were tested for this characteristic. These were inoculated with 150 infection-forming units (IFU of C. trachomatis A, B, Ba and C serovars. Growth was graded according to the number of IFUs per microscopic field (100X. A549-cell line was not susceptible to infection by any of the serovars. The growth of C. trachomatis was good to very good in McCoy and HeLa cell lines. Vero, BHK-21 and HEp-2 cell lines varied considerably in the susceptibility to infection.

  15. Bispecific Antibodies that Mediate Killing of Cells Infected with Human Immunodeficiency Virus of Any Strain

    Science.gov (United States)

    Berg, Jorg; Lotscher, Erika; Steimer, Kathelyn S.; Capon, Daniel J.; Baenziger, Jurg; Jack, Hans-Martin; Wabl, Matthias

    1991-06-01

    Although AIDS patients lose human immunodeficiency virus (HIV)-specific cytotoxic T cells, their remaining CD8-positive T lymphocytes maintain cytotoxic function. To exploit this fact we have constructed bispecific antibodies that direct cytotoxic T lymphocytes of any specificity to cells that express gp120 of HIV. These bispecific antibodies comprise one heavy/light chain pair from an antibody to CD3, linked to a heavy chain whose variable region has been replaced with sequences from CD4 plus a second light chain. CD3 is part of the antigen receptor on T cells and is responsible for signal transduction. In the presence of these bispecific antibodies, T cells of irrelevant specificity effectively lyse HIV-infected cells in vitro.

  16. A new strain of Streptomyces avermitilis produces high yield of oligomycin A with potent anti-tumor activity on human cancer cell lines in vitro.

    Science.gov (United States)

    Lin, Xiuping; Wen, Ying; Li, Meng; Chen, Zhi; Guo, Jia; Song, Yuan; Li, Jilun

    2009-01-01

    A new actinomycete strain, isolated from soil in China, strongly inhibited in vitro proliferation of human hepatoma, chronic myelogenous leukemia, and colonic carcinoma cell lines. The strain, designated L033, was identified as a strain of Streptomyces avermitilis based on cultural property, morphology, carbon source utilization, 16s rRNA gene analysis, and DNA-DNA relatedness studies. The anticancer component from L033 was purified to homogeneity by preparative positive-phase high-performance liquid chromatography and crystallization. Nuclear magnetic resonance and mass spectrometric analysis showed that this compound had the same structure as oligomycin A. Different with other reported naturally occurring strains of S. avermitilis, L033 produced high quantity of oligomycin A (maximal 1,461 microg/ml). Therefore, L033 was considered of great potential as an industrial oligomycin-A-producing strain.

  17. Nitric oxide (NO) production in mammalian non-tumorigenic epithelial cells of the small intestine and macrophages induced by individual strains of lactobacilli and bifidobacteria

    DEFF Research Database (Denmark)

    Pipenbaher, Natasa; Møller, Peter Lange; Dolinsek, Jan

    2009-01-01

    a significant increase in NO production compared with the control cells in the macrophage cell line 3D4/21. Results support the protective role of the individual strains of the genera Lactobacillus and Bifidobacterium and may lead to new approaches for manipulating and regulating immune responses at the mucosal...

  18. The civRT operon is important for Campylobacter jejuni strain 81-176 host cell interactions through regulation of the formate dehydrogenase operon

    Science.gov (United States)

    C. jejuni colonizes the intestinal mucosa, and the severity of disease in different strains is correlated with host cell interaction and invasion. A microarray screen to identify genes differentially regulated during C. jejuni interaction with tissue culture cells revealed the up-regulation of a two...

  19. Deficient recovery from potentially lethal damage in some gamma-irradiated human fibroblast cell strains

    Energy Technology Data Exchange (ETDEWEB)

    Arlett, C.F.; Priestley, A. (Medical Research Council, Brighton (UK). Cell Mutation Unit)

    1984-01-01

    The repair of potentially lethal damage following treatment with gamma radiation was investigated in human fibroblasts held in a non-cycling state by maintenance in a medium containing 0.5% foetal calf serum. Normal cells were found to be competent in the repair of PLD. Ataxia-telangiectasia cells were deficient as was a heterozygote suggesting that a failure to repair PLD may make it possible to detect such heterozygotes. Fibroblasts from Huntington's disease patients were either slightly or no more sensitive than cells from normal individuals. Cultures from two individuals in the former class showed limited capacity to repair PLD but cells from the latter class were as competent as normals. Thus assays of radiosensitivity where conditions allow for the repair of PLD may maximise small differences in sensitivity. Cells taken from three patients suffering from Basal Cell Naevus Syndrome were also shown to be defective in the repair of PLD. The existence of such a defect may be related to the increased frequency of basal cell cancer observed in exposed fields following irradiation of such individuals.

  20. Thyrotropin Receptor and Membrane Interactions in FRTL-5 Thyroid Cell Strain in Microgravity

    Science.gov (United States)

    Albi, E.; Ambesi-Impiombato, F. S.; Peverini, M.; Damaskopoulou, E.; Fontanini, E.; Lazzarini, R.; Curcio, F.; Perrella, G.

    2011-01-01

    The aim of this work was to analyze the possible alteration of thyrotropin (TSH) receptors in microgravity, which could explain the absence of thyroid cell proliferation in the space environment. Several forms of the TSH receptor are localized on the plasma membrane associated with caveolae and lipid rafts. The TSH regulates the fluidity of the cell membrane and the presence of its receptors in microdomains that are rich in sphingomyelin and cholesterol. TSH also stimulates cyclic adenosine monophosphate (cAMP) accumulation and cell proliferation. Reported here are the results of an experiment in which the FRTL-5 thyroid cell line was exposed to microgravity during the Texus-44 mission (launched February 7, 2008, from Kiruna, Sweden). When the parabolic flight brought the sounding rocket to an altitude of 264km, the culture media were injected with or without TSH in the different samples, and weightlessness prevailed on board for 6 minutes and 19 seconds. Control experiments were performed, in parallel, in an onboard 1g centrifuge and on the ground in Kiruna laboratory. Cell morphology and function were analyzed. Results show that in microgravity conditions the cells do not respond to TSH treatment and present an irregular shape with condensed chromatin, a modification of the cell membrane with shedding of the TSH receptor in the culture medium, and an increase of sphingomyelin-synthase and Bax proteins. It is possible that real microgravity induces a rearrangement of specific sections of the cell membrane, which act as platforms for molecular receptors, thus influencing thyroid cell function in astronauts during space missions.

  1. Application of a genetically encoded biosensor for live cell imaging of L-valine production in pyruvate dehydrogenase complex-deficient Corynebacterium glutamicum strains.

    Science.gov (United States)

    Mustafi, Nurije; Grünberger, Alexander; Mahr, Regina; Helfrich, Stefan; Nöh, Katharina; Blombach, Bastian; Kohlheyer, Dietrich; Frunzke, Julia

    2014-01-01

    The majority of biotechnologically relevant metabolites do not impart a conspicuous phenotype to the producing cell. Consequently, the analysis of microbial metabolite production is still dominated by bulk techniques, which may obscure significant variation at the single-cell level. In this study, we have applied the recently developed Lrp-biosensor for monitoring of amino acid production in single cells of gradually engineered L-valine producing Corynebacterium glutamicum strains based on the pyruvate dehydrogenase complex-deficient (PDHC) strain C. glutamicum ΔaceE. Online monitoring of the sensor output (eYFP fluorescence) during batch cultivation proved the sensor's suitability for visualizing different production levels. In the following, we conducted live cell imaging studies on C. glutamicum sensor strains using microfluidic chip devices. As expected, the sensor output was higher in microcolonies of high-yield producers in comparison to the basic strain C. glutamicum ΔaceE. Microfluidic cultivation in minimal medium revealed a typical Gaussian distribution of single cell fluorescence during the production phase. Remarkably, low amounts of complex nutrients completely changed the observed phenotypic pattern of all strains, resulting in a phenotypic split of the population. Whereas some cells stopped growing and initiated L-valine production, others continued to grow or showed a delayed transition to production. Depending on the cultivation conditions, a considerable fraction of non-fluorescent cells was observed, suggesting a loss of metabolic activity. These studies demonstrate that genetically encoded biosensors are a valuable tool for monitoring single cell productivity and to study the phenotypic pattern of microbial production strains.

  2. Growth and cell-division in extensive (XDR) and extremely drug resistant (XXDR) tuberculosis strains: transmission and atomic force observation.

    Science.gov (United States)

    Farnia, Parissa; Mohammad, Reza Masjedi; Merza, Muayad Aghali; Tabarsi, Payam; Zhavnerko, Gennadii Konstantinovich; Ibrahim, Tengku Azmi; Kuan, Ho Oi; Ghanavei, Jalladein; Farnia, Poopak; Ranjbar, Reza; Poleschuyk, Nikolai Nikolaevich; Titov, Leonid Petrovich; Owlia, Parviz; Kazampour, Mehadi; Setareh, Mohammad; Sheikolslami, Muaryam; Migliori, Giovanni Battista; Velayati, Ali Akbar

    2010-09-30

    The ultra-structure of Mycobacterium tuberculosis (MTB) was examined by transmission electronic (TEM)) and atomic force microscopy (AFM). The study was performed to describe the morphology of susceptible, multidrug-resistant (MDR), extensively drug-resistant (XDR) and extremely drug-resistant tuberculosis isolates (XXDR-TB) during their exponential growth phase. Four types of cell division were observed and described. While three of them (symmetrical, asymmetrical and branching type) occurred in all isolates studied, the fourth one (adapted type) was seen only in XDR and XXDR-TB bacilli. In the fourth type of cell division, a rod shaped mother cell produced a small round shape bacillus (0.3-0.5 μm). These round cells were different from buds or polar division, but similar to terminal endospores without showing the typing heat resistance. Based on the present observation, we suggest that XDR-and XXDR-TB bacilli accommodate changes helping them to overcome the hostile environment. Viewed under AFM, the other frequently detected shapes in MTB isolates were oval, V, Y and multi-branching filaments. These shape variation confirmed pleomorphic phenomena in MTB populations and the specific features of pan-resistant strains.

  3. Bacillus amyloliquefaciens strains isolated from moisture-damaged buildings produced surfactin and a substance toxic to mammalian cells.

    Science.gov (United States)

    Mikkola, Raimo; Andersson, Maria A; Grigoriev, Pavel; Teplova, Vera V; Saris, Nils-Erik L; Rainey, Frederick A; Salkinoja-Salonen, Mirja S

    2004-04-01

    Fungicidic Bacillus amyloliquefaciens strains isolated from the indoor environment of moisture-damaged buildings contained heat-stable, methanol-soluble substances that inhibited motility of boar spermatozoa within 15 min of exposure and killed feline lung cells in high dilution in 1 day. Boar sperm cells lost motility, cellular ATP, and NADH upon contact to the bacterial extract (0.2 microg dry wt/ml). Two bioactive substances were purified from biomass of the fungicidal isolates. One partially characterized substance, 1,197 Da, was moderately hydrophobic and contained leucine, proline, serine, aspartic acid, glutamic acid and tyrosine, in addition to chromophore(s) absorbing at 365 nm. In boar sperm and human neural cells (Paju), the compound depolarized the transmembrane potentials of mitochondria (Delta Psi(m)) and the plasma membrane (Delta Psi(p)) after a 20-min exposure and formed cation-selective channels in lipid membranes, with a selectivity K(+):Na(+):Ca(2+) of 26:15:3.5. The other substance was identified as a plasma-membrane-damaging lipopeptide surfactin. Plate-grown biomass of indoor Bacillus amyloliquefaciens contained ca. 7% of dry weight of the two substances, 1,197 Da and surfactin, in a ratio of 1:6 (w:w). The in vitro observed simultaneous collapse of both cytosolic and mitochondrial ATP in the affected mammalian cell, induced by the 1,197-Da cation channel, suggests potential health risks for occupants of buildings contaminated with such toxins.

  4. Optimization of protocols for derivation of mouse embryonic stem cell lines from refractory strains, including the non obese diabetic mouse.

    Science.gov (United States)

    Davies, Timothy J; Fairchild, Paul J

    2012-07-01

    The derivation of pluripotent embryonic stem cells (ESCs) from a variety of genetic backgrounds remains a desirable objective in the generation of mice functionally deficient in genes of interest and the modeling of human disease. Nevertheless, disparity in the ease with which different strains of mice yield ESC lines has long been acknowledged. Indeed, the generation of bona fide ESCs from the non obese diabetic (NOD) mouse, a well-characterized model of human type I diabetes, has historically proved especially difficult to achieve. Here, we report the development of protocols for the derivation of novel ESC lines from C57Bl/6 mice based on the combined use of high concentrations of leukemia inhibitory factor and serum-replacement, which is equally applicable to fresh and cryo-preserved embryos. Further, we demonstrate the success of this approach using Balb/K and CBA/Ca mice, widely considered to be refractory strains. CBA/Ca ESCs contributed to the somatic germ layers of chimeras and displayed a very high competence at germline transmission. Importantly, we were able to use the same protocol for the derivation of ESC lines from nonpermissive NOD mice. These ESCs displayed a normal karyotype that was robustly stable during long-term culture, were capable of forming teratomas in vivo and germline competent chimeras after injection into recipient blastocysts. Further, these novel ESC lines efficiently formed embryoid bodies in vitro and could be directed in their differentiation along the dendritic cell lineage, thus illustrating their potential application to the generation of cell types of relevance to the pathogenesis of type I diabetes.

  5. Intestinal Microbial Dysbiosis and Colonic Epithelial Cell Hyperproliferation by Dietary α-Mangostin is Independent of Mouse Strain

    Directory of Open Access Journals (Sweden)

    Fabiola Gutierrez-Orozco

    2015-01-01

    Full Text Available Beverages and supplements prepared from mangosteen fruit are claimed to support gut health and immunity, despite the absence of supporting evidence from clinical trials. We recently reported that α-mangostin (α-MG, the most abundant xanthone in mangosteen fruit, altered the intestinal microbiome, promoted dysbiosis, and exacerbated colitis in C57BL/6J mice. The objective of this study was to determine whether induction of dysbiosis by dietary α-MG is limited to the C57BL/6J strain or represents a more generic response to chronic intake of the xanthone on the gut microbiota of mice. C3H, Balb/c, Nude FoxN1nu, and C57BL/6J mice, each demonstrating unique microbiomes, were fed standard diet or diet containing 0.1% α-MG for four weeks. Dietary α-MG significantly altered the cecal and colonic microbiota in all four strains of mice, promoting a reduction in generally assumed beneficial bacterial groups while increasing the abundance of pathogenic bacteria. Consumption of α-MG was associated with reduced abundance of Firmicutes and increased abundance of Proteobacteria. The abundance of Lachnospiraceae, Ruminococcaceae, and Lactobacillaceae was reduced in α-MG-fed mice, while that of Enterobacteriaceae and Enterococcaceae was increased. Dietary α-MG also was associated with increased proliferation of colonic epithelial cells, infiltration of immune cells, infiltration of immune cells and increased fluid content in stool. These results suggest that ingestion of pharmacologic doses of xanthones in mangosteen-containing supplements may adversely alter the gut microbiota and should be used with caution.

  6. In vitro proliferation of haemopoietic cells in the presence of adherent cell layers. I. Culture conditions and strain dependence

    NARCIS (Netherlands)

    Reimann, J.; Burger, H.

    1979-01-01

    The culture system, in which a marrow-derived adherent cell population, established in vitro, exerts a long-term promoting influence on proliferation of haemopoietic cells, is reproduced. Essential parameters of the system are investigated; it is confirmed that the system is critically dependent on

  7. Expression of recombinant M2 and M5 muscarinic receptors in the Sf9-baculovirus system%在Sf9昆虫细胞-杆状病毒系统中表达毒蕈碱型M2及M5受体重组突变体

    Institute of Scientific and Technical Information of China (English)

    牟男; 孙洪良; 郑建全; 王丽韫

    2011-01-01

    OBJECTIVE To study the expression of human muscarinic receptors ( M2 and M5 recombinant receptors in the baculovirus expression system.METHODS The mutation of human wild type M2 and M5 receptors was constructed by PCR or/and overlap PCR as follows: ① The putative glyeosylation residues Asp 2, 3, 6, and 9 were replaced with Asn to prevent molecular heterogeneity; ② The central part of the protease-susceptible third intracellular loop was deleted; ③ A hexa-histidine tag and a thrombin cleavage site were added at the C terminus for purification.The recombinant receptor gene was confirmed and amplified by PCR, and subcloned to baculovrius pFastBac 1 vector.Then the recombinant vector was co-transfected with the linearized virus DNA into sf9 cells by Lipofectamine.The recombinant M2 and M5 receptor protein was prepared and purified.The expression level of M2 and M5 receptors was evaluated by Western blotting, and pharmacological characteristics were confirmed by radio-legend binding assay.RESULTS The target DNA fragment of M2(1018 bp) and M5 (1041 bp) recombinant receptors was amplified by overlap PCR.The recombinant plasmid pfastbacl/M2 (M5 ) vector was successfully constructed, and transfected to Sf9.Vacuolus pathological changes were observed within cells compared to non-transfection of Sf9.The baculovirus particle protein was prepared and purified from these infected cells.The expression of M2/M5 was further confirmed by Western blotting.The specific binding character of recombinant M2/M5 receptors was detected by radio-legend binding assay.CONCLUSION The expression of M2 and M5 recombinant receptors in the baculovirus expression system will facilitate studies on new drugs from M receptor or genetic engineering.%目的 为乙酰胆碱毒蕈碱(M)受体亚型特异性的变构调节剂及基因工程的研究提供实验平台.方法 用PCR及搭桥PCR法对乙酰胆碱M2及M5受体作以下突变:①将N-糖基化位点Asp突变为Asn;②删除对蛋白酶敏

  8. Adipose progenitor cells increase fibronectin matrix strain and unfolding in breast tumors

    Science.gov (United States)

    Chandler, E. M.; Saunders, M. P.; Yoon, C. J.; Gourdon, D.; Fischbach, C.

    2011-02-01

    Increased stiffness represents a hallmark of breast cancer that has been attributed to the altered physicochemical properties of the extracellular matrix (ECM). However, the role of fibronectin (Fn) in modulating the composition and mechanical properties of the tumor-associated ECM remains unclear. We have utilized a combination of biochemical and physical science tools to evaluate whether paracrine signaling between breast cancer cells and adipose progenitor cells regulates Fn matrix assembly and stiffness enhancement in the tumor stroma. In particular, we utilized fluorescence resonance energy transfer imaging to map the molecular conformation and stiffness of Fn that has been assembled by 3T3-L1 preadipocytes in response to conditioned media from MDA-MB231 breast cancer cells. Our results reveal that soluble factors secreted by tumor cells promote Fn expression, unfolding, and stiffening by adipose progenitor cells and that transforming growth factor-β serves as a soluble cue underlying these changes. In vivo experiments using orthotopic co-transplantation of primary human adipose-derived stem cells and MDA-MB231 into SCID mice support the pathological relevance of our results. Insights gained by these studies advance our understanding of the role of Fn in mammary tumorigenesis and may ultimately lead to improved anti-cancer therapies.

  9. Dependence of lattice strain relaxation, absorbance, and sheet resistance on thickness in textured ZnO@B transparent conductive oxide for thin-film solar cell applications

    OpenAIRE

    2016-01-01

    The interplay of surface texture, strain relaxation, absorbance, grain size, and sheet resistance in textured, boron-doped ZnO (ZnO@B), transparent conductive oxide (TCO) materials of different thicknesses used for thin film, solar cell applications is investigated. The residual strain induced by the lattice mismatch and the difference in the thermal expansion coefficient for thicker ZnO@B is relaxed, leading to an increased surface texture, stronger absorbance, larger grain size, and lower s...

  10. Cell wall carbohydrates content of pathogenic Candida albicans strain morphological forms.

    Science.gov (United States)

    Staniszewska, Monika; Bondaryk, Małgorzata; Rabczenko, Daniel; Smoleńska-Sym, Gabriela; Kurzatkowski, Wiesław

    2013-01-01

    The study evaluated the cell wall carbohydrates fraction in blastoconidia grown in YEPD medium at 30 degrees C and in the conglomerate of true hyphae grown in human serum at 37 degrees C. The clinical isolate obtained from a child with widespread C. albicans infection was used in the study. The cells were broken with glass beads, centrifuged to harvest the cell wall followed by subjection to TFA hydrolysis and in the result of that released monosaccharides were detected by HPAEC-PAD. Both, serum and temperature conditions (37 degrees C) affected germination process influencing the cell wall carbohydrates content when incubation in serum was prolonged from 1 to 18 h. The mannan content of blastoconidia was almost twofold higher compared to filamentous forms (149.25 +/- 299.24 vs 77.26 +/- 122.07). The glucan content was threefold lower in blastoconidia compared to hyphae (251.86 +/- 243.44 vs 755.81 +/- 1299.30). The chitin level was fourfold lower in blastoconidia compared to filaments (23.86 +/- 54.09 vs 106.29 +/- 170.12). The reason for the differences in the carbohydrates content may be related to type of morphology induced in different environmental conditions. Among tested carbohydrates, glucan appeared to be present in appreciably larger amounts in both tested morphological fractions. The ultrastructure of the blastoconidial cell wall revealed striking differences compared to the hyphae indicating the carbohydrates content alterations for wall assembly during hyphal growth at alkaline pH and temp. 37 degrees C. The study provided evidence for the relationship between morphogenesis, cell-cell adhesion induced by serum and changes in the level of carbohydrates content.

  11. The Openness/Intellect Model on the M5-50: Supporting the Flexibility of IPIP-Based Instruments

    OpenAIRE

    Vuyk, M. Alexandra; University of Kansas; Ingram IV, Paul B.; University of Kansas; Multon, Karen D.; University of Kansas; Warlick, Craig A.; University of Kansas

    2016-01-01

    This study examined ways to improve fit and interpretive capacity of the M5-50, an IPIP-based personality instrument, using the Openness/Intellect model (O/I) given a history of poor performance of the M5-50 Openness scale (Author, 2013; Socha, Cooper, & McCord, 2010). With participants from Amazon’s MTurk (n = 305), theoretical models for the five-factor model, Openness as a 10-item single factor, and the O/I model were tested and fitted poorly After removing one problematic item, the O/I m...

  12. Evaluation of the significance of cell wall polymers in flax infected with a pathogenic strain of Fusarium oxysporum.

    Science.gov (United States)

    Wojtasik, Wioleta; Kulma, Anna; Dymińska, Lucyna; Hanuza, Jerzy; Czemplik, Magdalena; Szopa, Jan

    2016-03-22

    Fusarium oxysporum infection leads to Fusarium-derived wilt, which is responsible for the greatest losses in flax (Linum usitatissimum) crop yield. Plants infected by Fusarium oxysporum show severe symptoms of dehydration due to the growth of the fungus in vascular tissues. As the disease develops, vascular browning and leaf yellowing can be observed. In the case of more virulent strains, plants die. The pathogen's attack starts with secretion of enzymes degrading the host cell wall. The main aim of the study was to evaluate the role of the cell wall polymers in the flax plant response to the infection in order to better understand the process of resistance and develop new ways to protect plants against infection. For this purpose, the expression of genes involved in cell wall polymer metabolism and corresponding polymer levels were investigated in flax seedlings after incubation with Fusarium oxysporum. This analysis was facilitated by selecting two groups of genes responding differently to the infection. The first group comprised genes strongly affected by the infection and activated later (phenylalanine ammonia lyase and glucosyltransferase). The second group comprised genes which are slightly affected (up to five times) and their expression vary as the infection progresses. Fusarium oxysporum infection did not affect the contents of cell wall polymers, but changed their structure. The results suggest that the role of the cell wall polymers in the plant response to Fusarium oxysporum infection is manifested through changes in expression of their genes and rearrangement of the cell wall polymers. Our studies provided new information about the role of cellulose and hemicelluloses in the infection process, the change of their structure and the expression of genes participating in their metabolism during the pathogen infection. We also confirmed the role of pectin and lignin in this process, indicating the major changes at the mRNA level of lignin metabolism genes

  13. Pravastatin and simvastatin inhibit the adhesion, replication and proliferation of Toxoplasma gondii (RH strain) in HeLa cells.

    Science.gov (United States)

    Sanfelice, Raquel Arruda; da Silva, Suelen Santos; Bosqui, Larissa Rodrigues; Miranda-Sapla, Milena Menegazzo; Barbosa, Bellisa Freitas; Silva, Rafaela José; Ferro, Eloísa A Vieira; Panagio, Luciano Aparecido; Navarro, Italmar Teodorico; Bordignon, Juliano; Conchon-Costa, Ivete; Pavanelli, Wander Rogerio; Almeida, Ricardo Sergio; Costa, Idessania Nazareth

    2017-03-01

    The conventional treatment for toxoplasmosis with pyrimethamine and sulfadiazine shows toxic effects to the host, and it is therefore necessary to search for new drugs. Some studies suggest the use of statins, which inhibit cholesterol synthesis in humans and also the initial processes of isoprenoid biosynthesis in the parasite. Thus, the objective of this study was to evaluate the activity of the statins pravastatin and simvastatin in HeLa cells infected in vitro with the RH strain of T. gondii. HeLa cells (1×10(5)) were infected with T. gondii tachyzoites (5×10(5)) following two different treatment protocols. In the first protocol, T. gondii tachyzoites were pretreated with pravastatin (50 and 100μg/mL) and simvastatin (1.56 and 3.125μg/mL) for 30min prior to infection. In the second, HeLa cells were first infected (5×10(5)) with tachyzoites and subsequently treated with pravastatin and simvastatin for 24h at the concentrations noted above. Initially, we evaluated the cytotoxicity of drugs by the MTT assay, number of tachyzoites adhered to cells, number of infected cells, and viability of tachyzoites by trypan blue exclusion. The supernatant of the cell cultures was collected post-treatment for determination of the pattern of Th1/Th2/Th17 cytokines by cytometric bead array. There was no cytotoxicity to HeLa cells with 50 and 100μg/mL pravastatin and 1.56 and 3.125μg/mL simvastatin. There was no change in the viability of tachyzoites that received pretreatment. Regarding the pre- and post-treatment of the cells with pravastatin and simvastatin alone, there was a reduction in adhesion, invasion and proliferation of cells to T. gondii. As for the production of cytokines, we found that IL-6 and IL-17 were significantly reduced in cells infected with T. gondii and treated with pravastatin and simvastatin, when compared to control. Based on these results, we can infer that pravastatin and simvastatin alone possess antiproliferative effects on tachyzoites forms

  14. Reaction and strain engineering for improved stereo-selective whole-cell reduction of a bicyclic diketone.

    Science.gov (United States)

    Johanson, Ted; Carlquist, Magnus; Olsson, Cecilia; Rudolf, Andreas; Frejd, Torbjörn; Gorwa-Grauslund, Marie F

    2008-01-01

    Reduction of bicyclo[2.2.2]octane-2,6-dione to (1R, 4S, 6S)-6-hydroxy-bicyclo[2.2.2]octane-2-one by whole cells of Saccharomyces cerevisiae was improved using an engineered recombinant strain and process design. The substrate inhibition followed a Han-Levenspiel model showing an effective concentration window between 12 and 22 g/l, in which the activity was kept above 95%. Yeast growth stage, substrate concentration and a stable pH were shown to be important parameters for effective conversion. The over-expression of the reductase gene YDR368w significantly improved diastereoselectivity compared to previously reported results. Using strain TMB4110 expressing YDR368w in batch reduction with pH control, complete conversion of 40 g/l (290 mM) substrate was achieved with 97% diastereomeric excess (de) and >99 enantiomeric excess (ee), allowing isolation of the optically pure ketoalcohol in 84% yield.

  15. Inflammatory cytokine and microRNA responses of primary human dendritic cells cultured with Helicobacter pylori strains.

    Directory of Open Access Journals (Sweden)

    Anaïs eHocès De La Guardia

    2013-08-01

    Full Text Available Primary human dendritic cells (DC were used to explore the inflammatory effectors, including cytokines and microRNAs, regulated by Helicobacter pylori. In a 48 h ex-vivo co-culture system, both H. pylori B38 and B45 strains activated human DCs and promoted a strong inflammatory response characterized by the early production of pro-inflammatory TNF and IL-6 cytokines, followed by IL-10, IL-1ß and IL-23 secretion. IL-23 was the only cytokine dependent on the cag pathogenicity island status of the bacterial strains. DC activation and cytokine production were accompanied by an early miR-146a upregulation followed by a strong miR-155 induction, which mainly controlled TNFα production. These results pave the way for further investigations into the nature of H. pylori antigens and the subsequently activated signaling pathways involved in the inflammatory response to H. pylori infection, the deregulation of which may likely contribute to gastric lymphomagenesis.

  16. Three-dimensional ionic conduction in the strained electrolytes of solid oxide fuel cells

    Science.gov (United States)

    Han, Yupei; Zou, Minda; Lv, Weiqiang; Mao, Yiwu; Wang, Wei; He, Weidong

    2016-05-01

    Flexible power sources including fuel cells and batteries are the key to realizing flexible electronic devices with pronounced foldability. To understand the bending effects in these devices, theoretical analysis on three-dimensional (3-D) lattice bending is necessary. In this report, we derive a 3-D analytical model to analyze the effects of electrolyte crystal bending on ionic conductivity in flexible solid-state batteries/fuel cells. By employing solid oxide fuel cells as a materials' platform, the intrinsic parameters of bent electrolyte materials, including lattice constant, Young's modulus, and Poisson ratio, are evaluated. Our work facilitates the rational design of highly efficient flexible electrolytes for high-performance flexible device applications.

  17. Listeria monocytogenes Strains Underrepresented during Selective Enrichment with an ISO Method Might Dominate during Passage through Simulated Gastric Fluid and In Vitro Infection of Caco-2 Cells

    Science.gov (United States)

    Zilelidou, Evangelia; Karmiri, Christina-Vasiliki; Zoumpopoulou, Georgia; Mavrogonatou, Eleni; Kletsas, Dimitris; Tsakalidou, Effie; Papadimitriou, Konstantinos; Drosinos, Eleftherios

    2016-01-01

    ABSTRACT Various Listeria monocytogenes strains may contaminate a single food product, potentially resulting in simultaneous exposure of consumers to multiple strains. However, due to bias in strain recovery, L. monocytogenes strains isolated from foods by selective enrichment (SE) might not always represent those that can better survive the immune system of a patient. We investigated the effect of cocultivation in tryptic soy broth with 0.6% yeast extract (TSB-Y) at 10°C for 8 days on (i) the detection of L. monocytogenes strains during SE with the ISO 11290-1:1996/Amd 1:2004 protocol and (ii) the in vitro virulence of strains toward the Caco-2 human colon epithelial cancer cell line following exposure to simulated gastric fluid (SGF; pH 2.0)-HCl (37°C). We determined whether the strains which were favored by SE would be effective competitors under the conditions of challenges related to gastrointestinal passage of the pathogen. Interstrain competition of L. monocytogenes in TSB-Y determined the relative population of each strain at the beginning of SE. This in turn impacted the outcome of SE (i.e., favoring survival of competitors with better fitness) and the levels exposed subsequently to SGF. However, strong growth competitors could be outcompeted after SGF exposure and infection of Caco-2 cells by strains outgrown in TSB-Y and underdetected (or even missed) during enrichment. Our data demonstrate a preferential selection of certain L. monocytogenes strains during enrichments, often not reflecting a selective advantage of strains during infection. These findings highlight a noteworthy scenario associated with the difficulty of matching the source of infection (food) with the L. monocytogenes isolate appearing to be the causative agent during listeriosis outbreak investigations. IMPORTANCE This report is relevant to understanding the processes involved in selection and prevalence of certain L. monocytogenes strains in different environments (i.e., foods or

  18. Phaeobacter sp. strain Y4I utilizes two separate cell-to-cell communication systems to regulate production of the antimicrobial indigoidine.

    Science.gov (United States)

    Cude, W Nathan; Prevatte, Carson W; Hadden, Mary K; May, Amanda L; Smith, Russell T; Swain, Caleb L; Campagna, Shawn R; Buchan, Alison

    2015-02-01

    The marine roseobacter Phaeobacter sp. strain Y4I synthesizes the blue antimicrobial secondary metabolite indigoidine when grown in a biofilm or on agar plates. Prior studies suggested that indigoidine production may be, in part, regulated by cell-to-cell communication systems. Phaeobacter sp. strain Y4I possesses two luxR and luxI homologous N-acyl-L-homoserine lactone (AHL)-mediated cell-to-cell communication systems, designated pgaRI and phaRI. We show here that Y4I produces two dominantAHLs, the novel monounsaturated N-(3-hydroxydodecenoyl)-L-homoserine lactone (3OHC(12:1)-HSL) and the relatively common N-octanoyl-L-homoserine lactone (C8-HSL), and provide evidence that they are synthesized by PhaI and PgaI, respectively.A Tn5 insertional mutation in either genetic locus results in the abolishment (pgaR::Tn5) or reduction (phaR::Tn5) of pigment production. Motility defects and denser biofilms were also observed in these mutant backgrounds, suggesting an overlap in the functional roles of these systems. Production of the AHLs occurs at distinct points during growth on an agar surface and was determined by isotope dilution high-performance liquid chromatography–tandem mass spectrometry (ID-HPLC-MS/MS) analysis.Within 2 h of surface inoculation, only 3OHC(12:1)-HSL was detected in agar extracts. As surface-attached cells became established (at approximately 10 h), the concentration of 3OHC(12:1)-HSL decreased, and the concentration of C8-HSL increased rapidly over 14 h.After longer (>24-h) establishment periods, the concentrations of the two AHLs increased to and stabilized at approximately 15 nM and approximately 600 nM for 3OHC12:1-HSL and C8-HSL, respectively. In contrast, the total amount of indigoidine increased steadily from undetectable to 642 Mby 48 h. Gene expression profiles of the AHL and indigoidine synthases (pgaI, phaI, and igiD) were consistent with their metabolite profiles. These data provide evidence that pgaRI and phaRI play overlapping roles

  19. A naturally occurring prfA truncation in a Listeria monocytogenes field strain contributes to reduced replication and cell-to-cell spread.

    Science.gov (United States)

    Rupp, Sebastian; Aguilar-Bultet, Lisandra; Jagannathan, Vidhya; Guldimann, Claudia; Drögemüller, Cord; Pfarrer, Christiane; Vidondo, Beatriz; Seuberlich, Torsten; Frey, Joachim; Oevermann, Anna

    2015-08-31

    Listeria (L.) monocytogenes is an environmental bacterium that may become an intracellular pathogen upon ingestion to cause gastroenteritis, septicaemia, abortions, and/or fatal infections of the central nervous system. We here describe a L. monocytogenes field strain (JF5171) isolated from a bovine placenta in the context of abortion, which exhibited attenuation in bovine brain-slice cultures. The whole genome of strain JF5171 was sequenced, and the invasion, replication, and intercellular spread of JF5171 were further analyzed by quantification of colony forming units and immunofluorescence studies. Phospholipase and hemolysis activity of JF5171 were also quantified along with transcription levels of actA, hly and prfA. The data obtained were compared to those of the widely used L. monocytogenes reference strain, EGD-e. JF5171 exhibited reduced replication and lower levels of phospholipase and hemolysis activity. Invasion and cell-to-cell spread was strongly decreased compared to EGD-e, and actin polymerization was absent. A frame shift deletion was identified in the JF5171 coding region of the major regulator for virulence, prfA. This resulted in a truncated C-terminus sequence (WEN* vs. WGKLN*). In addition, a point mutation resulted in a lysine to arginine substitution at amino acid position 197. Complementation with prfA from EGD-e and with (EGD-e) prfA-K197N increased the replication and spread efficiency of JF5171. In contrast, complementation with the truncated version of prfA had no effect. Taken together, these results suggest that the truncated C-terminus of prfA considerably contributes to the strongly attenuated phenotype observed in vitro. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. The Penicillium chrysogenum extracellular proteome. Conversion from a food-rotting strain to a versatile cell factory for white biotechnology.

    Science.gov (United States)

    Jami, Mohammad-Saeid; García-Estrada, Carlos; Barreiro, Carlos; Cuadrado, Abel-Alberto; Salehi-Najafabadi, Zahra; Martín, Juan-Francisco

    2010-12-01

    The filamentous fungus Penicillium chrysogenum is well-known by its ability to synthesize β-lactam antibiotics as well as other secondary metabolites. Like other filamentous fungi, this microorganism is an excellent host for secretion of extracellular proteins because of the high capacity of its protein secretion machinery. In this work, we have characterized the extracellular proteome reference map of P. chrysogenum Wisconsin 54-1255 by two-dimensional gel electrophoresis. This method allowed the correct identification of 279 spots by peptide mass fingerprinting and tandem MS. These 279 spots included 328 correctly identified proteins, which corresponded to 131 different proteins and their isoforms. One hundred and two proteins out of 131 were predicted to contain either classical or nonclassical secretion signal peptide sequences, providing evidence of the authentic extracellular location of these proteins. Proteins with higher representation in the extracellular proteome were those involved in plant cell wall degradation (polygalacturonase, pectate lyase, and glucan 1,3-β-glucosidase), utilization of nutrients (extracellular acid phosphatases and 6-hydroxy-d-nicotine oxidase), and stress response (catalase R). This filamentous fungus also secretes enzymes specially relevant for food industry, such as sulfydryl oxidase, dihydroxy-acid dehydratase, or glucoamylase. The identification of several antigens in the extracellular proteome also highlights the importance of this microorganism as one of the main indoor allergens. Comparison of the extracellular proteome among three strains of P. chrysogenum, the wild-type NRRL 1951, the Wis 54-1255 (an improved, moderate penicillin producer), and the AS-P-78 (a penicillin high-producer), provided important insights to consider improved strains of this filamentous fungus as versatile cell-factories of interest, beyond antibiotic production, for other aspects of white biotechnology.

  1. Attenuation of an original US porcine epidemic diarrhea virus strain PC22A via serial cell culture passage.

    Science.gov (United States)

    Lin, Chun-Ming; Hou, Yixuan; Marthaler, Douglas G; Gao, Xiang; Liu, Xinsheng; Zheng, Lanlan; Saif, Linda J; Wang, Qiuhong

    2017-03-01

    Although porcine epidemic diarrhea (PED) has caused huge economic losses in the pork industry worldwide, an effective live, attenuated vaccine is lacking. In this study, an original US, highly virulent PED virus (PEDV) strain PC22A was serially passaged in Vero CCL81 and Vero BI cells. The virus growth kinetics in cell culture, virulence in neonatal pigs and the whole genomic sequences of selected passages were examined. Increased virus titers and sizes of syncytia were observed at the 65th passage level (P65) and P120, respectively. Based on the severity of clinical signs, histopathological lesions and the distribution of PEDV antigens in the gut, the virulence of P100 and above, but not P95C13 (CCL81), was markedly reduced in 4-day-old, caesarian-derived, colostrum-deprived piglets. Subsequently, the attenuation of P120 and P160 was confirmed in 4-day-old, conventional suckling piglets. Compared with P120, P160 replicated less efficiently in the intestine of pigs and induced a lower rate of protection after challenge. Sequence analysis revealed that the virulent viruses [P3 and P95C13 (CCL81)] had one, one, sixteen (including an early termination of nine amino acids) and two amino acid differences in non-structure protein 1 (nsp1), nsp4, spike and membrane proteins, respectively, from the fully attenuated P160. However, the overall pattern of attenuation-related genetic changes in PC22A differed from those of the other four pairs of PEDV wild type strains and their attenuated derivatives. These results suggest that PEDV attenuation can occur through multiple molecular mechanisms. The knowledge provides insights into potential molecular mechanisms of PEDV attenuation. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Database of potential sources for earthquakes larger than M 5.5 in Italy. Version 2.0-2001

    Energy Technology Data Exchange (ETDEWEB)

    Valensise, G.; Pantatosti, D. [Istituto Nazionale di Geofisica e Vulcanologia, Rome (Italy)

    2001-07-01

    This volume presents and describes Version 2.0 of the Database of potential sources for earthquakes larger than M 5.5 in Italy (also referred to as DISS, the acronym of the short form database of Italy's seismogenic sources, or simply as database) that was first conceived at the Istituto Nazionale di Geofisica e Vulcanologia of Rome in 1996.

  3. Effect of Carburizing and Hardening Temperature on the Endurance of Forming Dies from Steel R6M5

    Science.gov (United States)

    Stepankin, I. N.; Ken'ko, V. M.; Boiko, A. A.

    2013-07-01

    Results of a study of the effect of carburizing of the forming surfaces of cold upset dies from high-speed steel R6M5 and of the hardening temperature on the structure and properties of the dies are presented. It is shown that the hardness and endurance of the carburized tools can be raised by hardening from lower temperatures.

  4. A Study of confinement for $Q\\bar{Q}$ potentials on D3, M2 & M5 branes

    CERN Document Server

    Quijada, Edward

    2016-01-01

    We study analytically and numerically the interaction potentials between a pair of quark an anti-quark on D3, M2 and M5 branes. These potentials are obtained using Maldacena's method involving Wilson loops and present confining and non-confining behaviours in different situations that we explore in this work.

  5. Curli Temper Adherence of Escherichia coli O157:H7 to Squamous Epithelial Cells from the Bovine Recto-Anal Junction in a Strain-Dependent Manner.

    Science.gov (United States)

    Kudva, Indira T; Carter, Michelle Q; Sharma, Vijay K; Stasko, Judith A; Giron, Jorge A

    2017-01-01

    Our recent studies have shown that intimin and the locus of enterocyte effacement-encoded proteins do not play a role in Escherichia coli O157:H7 (O157) adherence to the bovine recto-anal junction squamous epithelial (RSE) cells. To define factors that play a contributory role, we investigated the role of curli, fimbrial adhesins commonly implicated in adherence to various fomites and plant and human epithelial cells, in O157 adherence to RSE cells. Specifically, we examined (i) wild-type strains of O157; (ii) curli variants of O157 strains; (iii) isogenic curli deletion mutants of O157; and (iv) adherence inhibition of O157 using anti-curlin sera. Results of these experiments conducted under stringent conditions suggest that curli do not solely contribute to O157 adherence to RSE cells and in fact demonstrate a modulating effect on O157 adherence to RSE cells in contrast to HEp-2 cells (human epidermoid carcinoma of the larynx cells with HeLa contamination). The absence of curli and presence of blocking anti-curli antibodies enhanced O157-RSE cell interactions among some strains, thus alluding to a spatial, tempering effect of curli on O157 adherence to RSE cells when present. At the same time, the presence or absence of curli did not alter RSE cell adherence patterns of another O157 strain. These observations are at variance with the reported role of curli in O157 adherence to human cell lines such as HEp-2 and need to be factored in when developing anti-adherence modalities for preharvest control of O157 in cattle.

  6. A proteomic study of the differential protein expression in MDBK cells after bovine herpesvirus type 1 infection (BHV-1) strain treatment.

    Science.gov (United States)

    Guo, Li; Yang, Yanling; Liu, Linna; Liao, Peng; Wen, Yongjun; Wu, Hua; Cheng, Shipeng

    2015-01-01

    Different BHV-1 strains, such as the virulent IBRV LN01/08 strains and the attenuated vaccine strain IBRV LNM, produces different clinical immune responses; however, the study of the differential protein expression in Madin-Darby bovine kidney (MDBK) cells after BHV-1-infection still remains unclear. Here, we applied a comparative proteomic strategy, based on 2D and MALDI-TOF/MS platforms, to examine the differential expression of proteins in MDBK cells that were treated and not treated with virulent IBRV LN01/08 and attenuated IBRV LNM strains. A total of eight differential proteins, including pyruvate kinase, heat shock protein (HSP) 90 (HSP90AA1 and HSP90AB1), annexin A, albumin (ALB), scinderin (SCIN), tubulin (alpha 1a) and vimentin (VIM), were identified. Among these proteins, pyruvate kinase, and HSP90 (HSP90AB1), tubulin and vimentin were identified in the virulent IBRV LN01/08 strain group, but were not identified in the attenuated IBRV LNM group. These results play an important role in tumor formation and development, cell migration, tumor cell line apoptosis, cell invasion and viral infection. The HSP90 (HSP90AA1) protein was identified in the control group and the attenuated IBRV LNM-infected group. Most studies have shown that HSP90 proteins were more of a cancer gene target, and inhibiting its function would result to oncogene degradation during cancer treatment. On the other hand, ALB is associated to cell differentiation, apoptosis, necrosis, cell death, viral infection, autophagy, interstitial tissue inflammation, and cell survival. These results provide a theoretical basis for the systematic understanding of BHV-1-infection mechanisms and BHV-1-induced immune responses.

  7. Vpu-Deficient HIV Strains Stimulate Innate Immune Signaling Responses in Target Cells

    OpenAIRE

    Doehle, Brian P.; Chang, Kristina; Fleming, Lamar; McNevin, John; Hladik, Florian; McElrath, M. Juliana; Gale, Michael

    2012-01-01

    Acute virus infection induces a cell-intrinsic innate immune response comprising our first line of immunity to limit virus replication and spread, but viruses have developed strategies to overcome these defenses. HIV-1 is a major public health problem; however, the virus-host interactions that regulate innate immune defenses against HIV-1 are not fully defined. We have recently identified the viral protein Vpu to be a key determinant responsible for HIV-1 targeting and degradation of interfer...

  8. Mushroom body miscellanea: transgenic Drosophila strains expressing anatomical and physiological sensor proteins in Kenyon cells.

    Science.gov (United States)

    Pech, Ulrike; Dipt, Shubham; Barth, Jonas; Singh, Priyanka; Jauch, Mandy; Thum, Andreas S; Fiala, André; Riemensperger, Thomas

    2013-01-01

    The fruit fly Drosophila melanogaster represents a key model organism for analyzing how neuronal circuits regulate behavior. The mushroom body in the central brain is a particularly prominent brain region that has been intensely studied in several insect species and been implicated in a variety of behaviors, e.g., associative learning, locomotor activity, and sleep. Drosophila melanogaster offers the advantage that transgenes can be easily expressed in neuronal subpopulations, e.g., in intrinsic mushroom body neurons (Kenyon cells). A number of transgenes has been described and engineered to visualize the anatomy of neurons, to monitor physiological parameters of neuronal activity, and to manipulate neuronal function artificially. To target the expression of these transgenes selectively to specific neurons several sophisticated bi- or even multipartite transcription systems have been invented. However, the number of transgenes that can be combined in the genome of an individual fly is limited in practice. To facilitate the analysis of the mushroom body we provide a compilation of transgenic fruit flies that express transgenes under direct control of the Kenyon-cell specific promoter, mb247. The transgenes expressed are fluorescence reporters to analyze neuroanatomical aspects of the mushroom body, proteins to restrict ectopic gene expression to mushroom bodies, or fluorescent sensors to monitor physiological parameters of neuronal activity of Kenyon cells. Some of the transgenic animals compiled here have been published already, whereas others are novel and characterized here for the first time. Overall, the collection of transgenic flies expressing sensor and reporter genes in Kenyon cells facilitates combinations with binary transcription systems and might, ultimately, advance the physiological analysis of mushroom body function.

  9. Dependence of lattice strain relaxation, absorbance, and sheet resistance on thickness in textured ZnO@B transparent conductive oxide for thin-film solar cell applications.

    Science.gov (United States)

    Kou, Kuang-Yang; Huang, Yu-En; Chen, Chien-Hsun; Feng, Shih-Wei

    2016-01-01

    The interplay of surface texture, strain relaxation, absorbance, grain size, and sheet resistance in textured, boron-doped ZnO (ZnO@B), transparent conductive oxide (TCO) materials of different thicknesses used for thin film, solar cell applications is investigated. The residual strain induced by the lattice mismatch and the difference in the thermal expansion coefficient for thicker ZnO@B is relaxed, leading to an increased surface texture, stronger absorbance, larger grain size, and lower sheet resistance. These experimental results reveal the optical and material characteristics of the TCO layer, which could be useful for enhancing the performance of solar cells through an optimized TCO layer.

  10. Cytokine responses in primary chicken embryo intestinal cells infected with Campylobacter jejuni strains of human and chicken origin and the expression of bacterial virulence-associated genes

    DEFF Research Database (Denmark)

    Li, Yiping; Ingmer, Hanne; Madsen, Mogens

    2008-01-01

    of the bacterial genes. We have investigated the invasiveness of primary chicken embryo intestinal cells (CEICs) by C. jejuni strains of human and chicken origins and the production of pro-inflammatory cytokines as well as the expression of the bacterial virulence-associated genes during co-cultivation. Results C...... in vitro culture condition C. jejuni strains of both human and chicken origins can invade avian host cells with a pro-inflammatory response and that the virulence-associated genes of C. jejuni may play a role in this process....

  11. Ultrastructural study of adherence to and penetration of cultured cells by two invasive Escherichia coli strains isolated from infants with enteritis.

    OpenAIRE

    1984-01-01

    The adherence of invasive Escherichia coli strains 444-3 and 469-3 to human erythrocytes and to cultured HeLa and HEp-2 cells has been examined by electron microscopy. Bacteria elaborating type 1 fimbriae, glycocalyces , and nonfimbrial mannose-resistant hemagglutinins specific for human erythrocytes were identified in cultures of both strains, and each of these different bacterial surface components appeared to be involved in attachment of 444-3 and 469-3 to cultured epithelial cells or huma...

  12. Dependence of lattice strain relaxation, absorbance, and sheet resistance on thickness in textured ZnO@B transparent conductive oxide for thin-film solar cell applications

    Directory of Open Access Journals (Sweden)

    Kuang-Yang Kou

    2016-01-01

    Full Text Available The interplay of surface texture, strain relaxation, absorbance, grain size, and sheet resistance in textured, boron-doped ZnO (ZnO@B, transparent conductive oxide (TCO materials of different thicknesses used for thin film, solar cell applications is investigated. The residual strain induced by the lattice mismatch and the difference in the thermal expansion coefficient for thicker ZnO@B is relaxed, leading to an increased surface texture, stronger absorbance, larger grain size, and lower sheet resistance. These experimental results reveal the optical and material characteristics of the TCO layer, which could be useful for enhancing the performance of solar cells through an optimized TCO layer.

  13. Proteomics profile changes in cisplatin-treated human ovarian cancer cell strain

    Institute of Scientific and Technical Information of China (English)

    LI Zhengyu; ZHAO Xia; YANG Jinliang; WEI Yuquan

    2005-01-01

    To compare the alterations in proteomes between cisplatin-treated and -untreated human ovarian cancer SKOV3 cells, and to explore the feasibility of proteomics in research about antitumor mechanisms of agents, SKOV3 cells were exposed to cisplatin (6 μg/mL) for 6 h. Then, the cells were collected and solubilized and global proteins were extracted by lysis buffer; two-dimensional electrophoresis was conducted with the IPG readystrips as carriers; the gels were stained with Coomassie blue and alterations between gels were compared by PDQuest. Eventually, 11 spots with significant differences were selected and excised and the proteins were identified by PMF and MS/MS analysis. The results revealed that exposure to cisplatin could notably increase expressions of some proteins, such as tropomyosin family, actin family, triosephosphate isomerase family, and HSP60, etc.; while expressions of some other proteins decreased, such as enolase family, etc. Those proteins were involved in cellular energy metabolism, transformation, apoptosis and morphologic maintenance, which suggested that alterations of those physiological processes might be involved in anti-tumor mechanism of cisplatin.

  14. Characterization of two strains of Anaplasma marginale isolated from cattle in Rio de Janeiro, Brazil, after propagation in tick cell culture.

    Science.gov (United States)

    Baêta, Bruna A; Ribeiro, Carla C D U; Teixeira, Rafaella C; Cabezas-Cruz, Alejandro; Passos, Lygia M F; Zweygarth, Erich; Fonseca, Adivaldo H

    2015-03-01

    IDE8 tick cell cultures have been used for the isolation and propagation of several isolates of Anaplasma marginale. The genetic heterogeneity of A. marginale strains in cattle is diverse in endemic regions worldwide and the analyses of msp1α (major surface protein 1 alpha) gene sequences have allowed the identification of different strains. This study reports the isolation and propagation of two new isolates of A. marginale in IDE8 cells from blood of two cattle and their morphological and molecular characterization using light microscopy and the msp1α gene, respectively. Small colonies were observed in cytospin smears of each of the isolates 60 days after culture initiation. Based on msp1α sequence variation, the two isolates were found to be separate strains and were named AmRio1 and AmRio2. Analysis of msp1α microsatellite in both strains resulted in a single genotype, genotype E. The amino acid sequence of one MSP1α tandem repeat from the strain AmRio1 resulted in a new sequence (named 162) with one amino acid change. The results of these phylogenetic analyses demonstrated that A. marginale strains from Brazil and Argentina formed two large clusters of which one was less divergent that the other.

  15. Different Transcriptional Profiles of Human Monocyte-Derived Dendritic Cells Infected with Distinct Strains of Mycobacterium tuberculosis and Mycobacterium bovis Bacillus Calmette-Guérin

    Directory of Open Access Journals (Sweden)

    Nunzia Sanarico

    2011-01-01

    Full Text Available In order to analyze dendritic cells (DCs activation following infection with different mycobacterial strains, we studied the expression profiles of 165 genes of human monocyte-derived DCs infected with H37Rv, a virulent Mycobacterium tuberculosis (MTB laboratory strain, CMT97, a clinical MTB isolate, Mycobacterium bovis bacillus Calmette-Guérin (BCG, Aventis Pasteur, and BCG Japan, both employed as vaccine against tuberculosis. The analysis of the gene expression reveals that, despite a set of genes similarly modulated, DCs response resulted strain dependent. In particular, H37Rv significantly upregulated EBI3 expression compared with BCG Japan, while it was the only strain that failed to release a significant IL-10 amount. Of note, BCG Japan showed a marked increase in CCR7 and TNF-α expression regarding both MTB strains and it resulted the only strain failing in exponential intracellular growth. Our results suggest that DCs display the ability to elicit a tailored strain-specific immune response.

  16. Phenotypic and genotypic characteristics of novel mouse cell line (NIH/3T3-adapted human enterovirus 71 strains (EV71:TLLm and EV71:TLLmv.

    Directory of Open Access Journals (Sweden)

    Carla Bianca Luena Victorio

    Full Text Available Since its identification in 1969, Enterovirus 71 (EV71 has been causing periodic outbreaks of infection in children worldwide and most prominently in the Asia-Pacific Region. Understanding the pathogenesis of Enterovirus 71 (EV71 is hampered by the virus's inability to infect small animals and replicate in their derived in vitro cultured cells. This manuscript describes the phenotypic and genotypic characteristics of two selected EV71 strains (EV71:TLLm and EV71:TLLmv, which have been adapted to replicate in mouse-derived NIH/3T3 cells, in contrast to the original parental virus which is only able to replicate in primate cell lines. The EV71:TLLm strain exhibited productive infection in all primate and rodent cell lines tested, while EV71:TLLmv exhibited greater preference for mouse cell lines. EV71:TLLmv displayed higher degree of adaptation and temperature adaptability in NIH/3T3 cells than in Vero cells, suggesting much higher fitness in NIH/3T3 cells. In comparison with the parental EV71:BS strain, the adapted strains accumulated multiple adaptive mutations in the genome resulting in amino acid substitutions, most notably in the capsid-encoding region (P1 and viral RNA-dependent RNA polymerase (3D. Two mutations, E167D and L169F, were mapped to the VP1 canyon that binds the SCARB2 receptor on host cells. Another two mutations, S135T and K140I, were located in the VP2 neutralization epitope spanning amino acids 136-150. This is the first report of human EV71 with the ability to productively infect rodent cell lines in vitro.

  17. A single methyltransferase YefA (RlmCD) catalyses both m5U747 and m5U1939 modifications in Bacillus subtilis 23S rRNA

    DEFF Research Database (Denmark)

    Desmolaize, Benoit; Fabret, Céline; Brégeon, Damien;

    2011-01-01

    . However, as previously shown, the m(5)U54 modification in B. subtilis tRNAs is catalysed in a fundamentally different manner by the folate-dependent enzyme TrmFO, which is unrelated to the E. coli TrmA. Here, we show that methylation of U747 and U1939 in B. subtilis rRNA is catalysed by a single enzyme...

  18. Comparative growth of spotted fever group Rickettsia spp. strains in Vero cells

    Science.gov (United States)

    Silva, Arannadia Barbosa; Duarte, Myrian Morato; Vizzoni, Vinicius Figueiredo; Duré, Ana Íris de Lima; Lopéz, Diego Montenegro; Nogueira, Rita de Maria Seabra; Soares, Carlos Augusto Gomes; Machado-Ferreira, Erik; Gazêta, Gilberto Salles

    2016-01-01

    In Brazil, the spotted fever group (SFG) Rickettsia rickettsii and Rickettsia parkeri related species are the etiological agents of spotted fever rickettsiosis. However, the SFG, Rickettsia rhipicephali, that infects humans, has never been reported. The study of growth dynamics can be useful for understanding the infective and invasive capacity of these pathogens. Here, the growth rates of the Brazilian isolates R. rickettsii str. Taiaçu, R. parkeri str. At#24, and R. rhipicephali HJ#5, were evaluated in Vero cells by quantitative polymerase chain reaction. R. rhipicephali showed different kinetic growth compared to R. rickettsii and R. parkeri. PMID:27508322

  19. Combined Effects of Mechanical Strain and Hydroxyapatite/Collagen Composite on Osteogenic Differentiation of Rat Bone Marrow Derived Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Yan Huang

    2013-01-01

    Full Text Available Mesenchymal stem cells (MSCs represent a promising source for bone repair and regeneration. Recent lines of evidence have shown that appropriate strain could regulate the osteogenic differentiation of MSCs. Our previous study demonstrated that hydroxyapatite/collagen (HA/Col composite also played an important role in the osteogenic differentiation of MSCs. The aim of this study is to investigate the effects of mechanical strain and HA/Col composite on the osteogenic differentiation of rat bone marrow derived MSCs (rBMSCs in vitro. rBMSCs were treated with cyclic strain generated by a self-designed stretching device with or without the presence of HA/Col composite. Osteogenic differentiation levels were evaluated using reverse transcription polymerase chain reaction (RT-PCR, alkaline phosphatase spectrophotometry, and western blotting. The results demonstrated that mechanical strain combined with HA/Col composite could obviously induce the differentiation of rBMSCs into osteoblasts, which had a better effect than only mechanical strain or HA/Col composite treatment. This provides a new avenue for mechanistic studies of stem cell differentiation and a novel approach to obtain more committed differentiated cells.

  20. Comparison of Chloroflexus aurantiacus strain J-10-fl proteomes of cells grown chemoheterotrophically and photoheterotrophically

    Energy Technology Data Exchange (ETDEWEB)

    Cao, Li; Bryant, Donald A.; Schepmoes, Athena A.; Vogl, Kajetan; Smith, Richard D.; Lipton, Mary S.; Callister, Stephen J.

    2012-01-17

    Chloroflexus aurantiacus J-10-fl is a thermophilic green bacterium, a filamentous anoxygenic phototroph, and the model organism of the phylum Chloroflexi. We applied high-throughput, liquid chromatography-mass spectrometry in a global quantitative proteomics investigation of C. aurantiacus cells grown under oxic (chemoorganoheterotrophically) and anoxic (photoorganoheterotrophically) redox states. Our global analysis identified 13,524 high-confidence peptides that matched to 1,286 annotated proteins, 242 of which were either uniquely identified or significantly increased in abundance under anoxic culture conditions. Fifty-three of the 242 proteins are previously characterized photosynthesis-related proteins, including chlorosome proteins, proteins involved in the bacteriochlorophyll biosynthesis, 3-hydroxypropionate (3-OHP) CO2 fixation pathway, and components of electron transport chains. The remaining 190 proteins have not previously been reported. Of these, five proteins were found to be encoded by genes from a novel operon and observed only in photoheterotrophically grown cells. These proteins candidates may prove useful in further deciphering the phototrophic physiology of C. aurantiacus and other filamentous anoxygenic phototrophs.

  1. Comparative evaluation of cell culture-adapted and chicken embryo-adapted fowl pox vaccine strains.

    Science.gov (United States)

    Baxi, M K; Oberoi, M S

    1999-01-01

    Two types of vaccines, chicken embryo adapted (VacCE) and cell culture adapted (VacCC), were tested for their efficacy to elicite the immune response in birds vaccinated at 2 and 8 wk of age. The cell-mediated immune response studied by blastogenesis assay showed that birds vaccinated at the second week of age by both VacCE and VacCC vaccines had significant increase in T-lymphocyte count at 21 days postvaccination (PV) and 7 days postchallenge (PC), whereas in birds vaccinated at 8 wk of age, a significant increase was seen at 21 days PV and 7 days PC with the VacCC vaccine. The rise in passive hemagglutination titers was observed up to 21 days PV and 7 days PC in birds vaccinated at 2 wk of age. However, only the birds vaccinated with VacCC at 8 wk of age showed rise in titers at days 21 PV and 7 PC. Birds were challenged 90 days PV by scarification on the thigh region, and the birds vaccinated with VacCC showed 90% and 70% protection when vaccinated at 2 and 8 wk, respectively. The birds vaccinated with VacCE showed only 60% and 20% protection at the corresponding levels, respectively.

  2. Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells

    Science.gov (United States)

    Shi, Wen; Jia, Shuo; Zhao, Haiyuan; Yin, Jiyuan; Wang, Xiaona; Yu, Meiling; Ma, Sunting; Wu, Yang; Chen, Ying; Fan, Wenlu; Xu, Yigang; Li, Yijing

    2017-01-01

    Porcine epidemic diarrhea virus (PEDV), which is the causative agent of porcine epidemic diarrhea in China and other countries, is responsible for serious economic losses in the pork industry. Inactivated PEDV vaccine plays a key role in controlling the prevalence of PEDV. However, consistently low viral titers are obtained during the propagation of PEDV in vitro; this represents a challenge to molecular analyses of the virus and vaccine development. In this study, we successfully isolated a PEDV isolate (strain NJ) from clinical samples collected during a recent outbreak of diarrhea in piglets in China, using porcine intestinal epithelial cells (IEC). We found that the isolate was better adapted to growth in IECs than in Vero cells, and the titer of the IEC cultures was 104.5 TCID50/0.1 mL at passage 45. Mutations in the S protein increased with the viral passage and the mutations tended towards attenuation. Viral challenge showed that the survival of IEC-adapted cultures was higher at the 45th passage than at the 5th passage. The use of IECs to isolate and propagate PEDV provides an effective approach for laboratory-based diagnosis of PEDV, as well as studies of the epidemiological characteristics and molecular biology of this virus. PMID:28117718

  3. Mitochondrial proteomics of the acetic acid - induced programmed cell death response in a highly tolerant Zygosaccharomyces bailii - derived hybrid strain

    Science.gov (United States)

    Guerreiro, Joana F.; Sampaio-Marques, Belém; Soares, Renata; Coelho, Ana V.; Leão, Cecília; Ludovico, Paula; Sá-Correia, Isabel

    2016-01-01

    Very high concentrations of acetic acid at low pH induce programmed cell death (PCD) in both the experimental model Saccharomyces cerevisiae and in Zygosaccharomyces bailii, the latter being considered the most problematic acidic food spoilage yeast due to its remarkable intrinsic resistance to this food preservative. However, while the mechanisms underlying S. cerevisiae PCD induced by acetic acid have been previously examined, the corresponding molecular players remain largely unknown in Z. bailii. Also, the reason why acetic acid concentrations known to be necrotic for S. cerevisiae induce PCD with an apoptotic phenotype in Z. bailii remains to be elucidated. In this study, a 2-DE-based expression mitochondrial proteomic analysis was explored to obtain new insights into the mechanisms involved in PCD in the Z. bailii derived hybrid strain ISA1307. This allowed the quantitative assessment of expression of protein species derived from each of the parental strains, with special emphasis on the processes taking place in the mitochondria known to play a key role in acetic acid - induced PCD. A marked decrease in the content of proteins involved in mitochondrial metabolism, in particular, in respiratory metabolism (Cor1, Rip1, Lpd1, Lat1 and Pdb1), with a concomitant increase in the abundance of proteins involved in fermentation (Pdc1, Ald4, Dld3) was registered. Other differentially expressed identified proteins also suggest the involvement of the oxidative stress response, protein translation, amino acid and nucleotide metabolism, among other processes, in the PCD response. Overall, the results strengthen the emerging concept of the importance of metabolic regulation of yeast PCD. PMID:28357336

  4. Isolation and characterization of a novel strain of mesenchymal stem cells from mouse umbilical cord: potential application in cell-based therapy.

    Directory of Open Access Journals (Sweden)

    Wen-Wen Li

    Full Text Available Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs have recently been recognized as a potential source for cell-based therapy in various preclinical animal models, such as Parkinson's disease, cerebral ischemia, spinal cord injury, and liver failure; however, the precise cellular and molecular mechanisms underlying the beneficial outcomes remain under investigation. There is a growing concern regarding rejection and alteration of genetic code using this xenotransplantation approach. In this study, a novel strain of murine MSCs derived from the umbilical cord of wild-type and green fluorescent protein (GFP transgenic mice have been successfully isolated, expanded, and characterized. After 10 passages, the mUC-MSCs developed a rather homogeneous, triangular, spindle-shaped morphology, and were sub-cultured up to 7 months (over 50 passages without overt changes in morphology and doubling time. Cell surface markers are quite similar to MSCs isolated from other tissue origins as well as hUC-MSCs. These mUC-MSCs can differentiate into osteoblasts, adipocytes, neurons, and astrocytes in vitro, as well as hematopoietic lineage cells in vivo. mUC-MSCs also possess therapeutic potential against two disease models, focal ischemic stroke induced by middle cerebral artery occlusion (MCAo and acute hepatic failure. Subtle differences in the expression of cytokine-related genes exist between mUC-MSCs and hUC-MSCs, which may retard and jeopardize the advance of cell therapy. Allografts of these newly established mUC-MSCs into various mouse disease models may deepen our insights into the development of more effective cell therapy regimens.

  5. Coseismic and postseismic deformation due to the 2007 M5.5 Ghazaband fault earthquake, Balochistan, Pakistan

    Science.gov (United States)

    Fattahi, H.; Amelung, F.; Chaussard, E.; Wdowinski, S.

    2015-05-01

    Time series analysis of interferometric synthetic aperture radar data reveals coseismic and postseismic surface displacements associated with the 2007 M5.5 earthquake along the southern Ghazaband fault, a major but little studied fault in Pakistan. Modeling indicates that the coseismic surface deformation was caused by ~9 cm of strike-slip displacement along a shallow subvertical fault. The earthquake was followed by at least 1 year of afterslip, releasing ~70% of the moment of the main event, equivalent to a M5.4 earthquake. This high aseismic relative to the seismic moment release is consistent with previous observations for moderate earthquakes (M < 6) and suggests that smaller earthquakes are associated with a higher aseismic relative to seismic moment release than larger earthquakes.

  6. YebU is a m5C methyltransferase specific for 16 S rRNA nucleotide 1407

    DEFF Research Database (Denmark)

    Andersen, Niels Møller; Douthwaite, Stephen

    2006-01-01

    The rRNAs in Escherichia coli contain methylations at 24 nucleotides, which collectively are important for ribosome function. Three of these methylations are m5C modifications located at nucleotides C967 and C1407 in 16S rRNA and at nucleotide C1962 in 23S rRNA. Bacterial rRNA modifications gener...... methyltransferase gene rsmF, and that the nomenclature system be extended to include the rRNA methyltransferases that still await identification....

  7. Results of Severe Fuel Damage Experiment QUENCH-14 with Advanced Rod Cladding M5®. (KIT Scientific Reports ; 7549)

    OpenAIRE

    STUCKERT J.; Große, M.; Stegmaier, U.; Steinbrück, M.

    2010-01-01

    The QUENCH experiments are to investigate the hydrogen release resulting from the water injection into an uncovered core of a Light Water Reactor as well as the high-temperature behavior of core materials. The QUENCH-14 experiment investigated the effect of M5® cladding material on bundle oxidation and core reflood, in comparison with the tests QUENCH-06 that used standard Zircaloy-4 and QUENCH-12 that used VVER E110-claddings.

  8. Differential Effects of Escherichia coli Nissle and Lactobacillus rhamnosus Strain GG on Human Rotavirus Binding, Infection, and B Cell Immunity.

    Science.gov (United States)

    Kandasamy, Sukumar; Vlasova, Anastasia N; Fischer, David; Kumar, Anand; Chattha, Kuldeep S; Rauf, Abdul; Shao, Lulu; Langel, Stephanie N; Rajashekara, Gireesh; Saif, Linda J

    2016-02-15

    Rotavirus (RV) causes significant morbidity and mortality in children worldwide. The intestinal microbiota plays an important role in modulating host-pathogen interactions, but little is known about the impact of commonly used probiotics on human RV (HRV) infection. In this study, we compared the immunomodulatory effects of Gram-positive (Lactobacillus rhamnosus strain GG [LGG]) and Gram-negative (Escherichia coli Nissle [EcN]) probiotic bacteria on virulent human rotavirus (VirHRV) infection and immunity using neonatal gnotobiotic piglets. Gnotobiotic piglets were colonized with EcN, LGG, or EcN+LGG or uncolonized and challenged with VirHRV. Mean peak virus shedding titers and mean cumulative fecal scores were significantly lower in EcN-colonized compared with LGG-colonized or uncolonized piglets. Reduced viral shedding titers were correlated with significantly reduced small intestinal HRV IgA Ab responses in EcN-colonized compared with uncolonized piglets post-VirHRV challenge. However the total IgA levels post-VirHRV challenge in the intestine and pre-VirHRV challenge in serum were significantly higher in EcN-colonized than in LGG-colonized piglets. In vitro treatment of mononuclear cells with these probiotics demonstrated that EcN, but not LGG, induced IL-6, IL-10, and IgA, with the latter partially dependent on IL-10. However, addition of exogenous recombinant porcine IL-10 + IL-6 to mononuclear cells cocultured with LGG significantly enhanced IgA responses. The greater effectiveness of EcN in moderating HRV infection may also be explained by the binding of EcN but not LGG to Wa HRV particles or HRV 2/4/6 virus-like particles but not 2/6 virus-like particles. Results suggest that EcN and LGG differentially modulate RV infection and B cell responses.

  9. Convergent synthesis of a tetrasaccharide repeating unit of the O-specific polysaccharide from the cell wall lipopolysaccharide of Azospirillum brasilense strain Sp7

    Directory of Open Access Journals (Sweden)

    Pintu Kumar Mandal

    2014-01-01

    Full Text Available A straightforward convergent synthesis has been carried out for the tetrasaccharide repeating unit of the O-specific cell wall lipopolysaccharide of the strain Sp7 of Azospirillum brasilense. The target tetrasaccharide has been synthesized from suitably protected monosaccharide intermediates in 42% overall yield in seven steps by using a [2 + 2] block glycosylation approach.

  10. Assessment of the pathogenicity of cell-culture-adapted Newcastle disease virus strain Komarov.

    Science.gov (United States)

    Visnuvinayagam, Sivam; Thangavel, K; Lalitha, N; Malmarugan, S; Sukumar, Kuppannan

    2015-01-01

    Newcastle disease vaccines hitherto in vogue are produced from embryonated chicken eggs. Egg-adapted mesogenic vaccines possess several drawbacks such as paralysis and mortality in 2-week-old chicks and reduced egg production in the egg-laying flock. Owing to these possible drawbacks, we attempted to reduce the vaccine virulence for safe vaccination by adapting the virus in a chicken embryo fibroblast cell culture (CEFCC) system. Eighteen passages were carried out by CEFCC, and the pathogenicity was assessed on the basis of the mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index, at equal passage intervals. Although the reduction in virulence demonstrated with increasing passage levels in CEFCC was encouraging, 20% of the 2-week-old birds showed paralytic symptoms with the virus vaccine from the 18(th)(final) passage. Thus, a tissue-culture-adapted vaccine would demand a few more passages by CEFCC in order to achieve a complete reduction in virulence for use as a safe and effective vaccine, especially among younger chicks. Moreover, it can be safely administered even to unprimed 8-week-old birds.

  11. Assessment of the pathogenicity of cell-culture-adapted Newcastle disease virus strain Komarov

    Directory of Open Access Journals (Sweden)

    Sivam Visnuvinayagam

    2015-09-01

    Full Text Available Newcastle disease vaccines hitherto in vogue are produced from embryonated chicken eggs. Egg-adapted mesogenic vaccines possess several drawbacks such as paralysis and mortality in 2-week-old chicks and reduced egg production in the egg-laying flock. Owing to these possible drawbacks, we attempted to reduce the vaccine virulence for safe vaccination by adapting the virus in a chicken embryo fibroblast cell culture (CEFCC system. Eighteen passages were carried out by CEFCC, and the pathogenicity was assessed on the basis of the mean death time, intracerebral pathogenicity index, and intravenous pathogenicity index, at equal passage intervals. Although the reduction in virulence demonstrated with increasing passage levels in CEFCC was encouraging, 20% of the 2-week-old birds showed paralytic symptoms with the virus vaccine from the 18th(final passage. Thus, a tissue-culture-adapted vaccine would demand a few more passages by CEFCC in order to achieve a complete reduction in virulence for use as a safe and effective vaccine, especially among younger chicks. Moreover, it can be safely administered even to unprimed 8-week-old birds.

  12. The Wyckoff positional order and polyhedral intergrowth in the M3B2- and M5B3-type boride precipitated in the Ni-based superalloys

    Science.gov (United States)

    Hu, X. B.; Zhu, Y. L.; Sheng, N. C.; Ma, X. L.

    2014-12-01

    Ni-based single superalloys play a crucial role in the hottest parts of jet engines. However, due to the complex geometry and macro-segregation during the solidification process, the cast defect such as stray grains is inevitable. Therefore, the transient liquid phase (TLP) bonding which can join several small single crystalline castings together is gradually believed to be an effective method for improving the yields of production of the complex components. The melting point depressant element B is always added into the interlayer filler material. Consequently, borides including the M3B2 and M5B3 phase usually precipitate during the TLP bonding process. So a comprehensive knowledge of the fine structural characteristics of the borides is very critical for an accurate evaluation of the TLP bonding process. In this work, by means of the aberration-corrected transmission electron microscopy, we show, at an atomic scale, the Wyckoff positional order phenomenon of the metal atoms in the unit cell of M3B2- and M5B3-type boride. Meanwhile, the defect along the (001) plane of the above two types of boride are determined to be the polyhedral intergrowth with complex configurations.

  13. Drought forecasting in eastern Australia using multivariate adaptive regression spline, least square support vector machine and M5Tree model

    Science.gov (United States)

    Deo, Ravinesh C.; Kisi, Ozgur; Singh, Vijay P.

    2017-02-01

    Drought forecasting using standardized metrics of rainfall is a core task in hydrology and water resources management. Standardized Precipitation Index (SPI) is a rainfall-based metric that caters for different time-scales at which the drought occurs, and due to its standardization, is well-suited for forecasting drought at different periods in climatically diverse regions. This study advances drought modelling using multivariate adaptive regression splines (MARS), least square support vector machine (LSSVM), and M5Tree models by forecasting SPI in eastern Australia. MARS model incorporated rainfall as mandatory predictor with month (periodicity), Southern Oscillation Index, Pacific Decadal Oscillation Index and Indian Ocean Dipole, ENSO Modoki and Nino 3.0, 3.4 and 4.0 data added gradually. The performance was evaluated with root mean square error (RMSE), mean absolute error (MAE), and coefficient of determination (r2). Best MARS model required different input combinations, where rainfall, sea surface temperature and periodicity were used for all stations, but ENSO Modoki and Pacific Decadal Oscillation indices were not required for Bathurst, Collarenebri and Yamba, and the Southern Oscillation Index was not required for Collarenebri. Inclusion of periodicity increased the r2 value by 0.5-8.1% and reduced RMSE by 3.0-178.5%. Comparisons showed that MARS superseded the performance of the other counterparts for three out of five stations with lower MAE by 15.0-73.9% and 7.3-42.2%, respectively. For the other stations, M5Tree was better than MARS/LSSVM with lower MAE by 13.8-13.4% and 25.7-52.2%, respectively, and for Bathurst, LSSVM yielded more accurate result. For droughts identified by SPI ≤ - 0.5, accurate forecasts were attained by MARS/M5Tree for Bathurst, Yamba and Peak Hill, whereas for Collarenebri and Barraba, M5Tree was better than LSSVM/MARS. Seasonal analysis revealed disparate results where MARS/M5Tree was better than LSSVM. The results highlight the

  14. Lactobacillus plantarum Strains Can Enhance Human Mucosal and Systemic Immunity and Prevent Non-steroidal Anti-inflammatory Drug Induced Reduction in T Regulatory Cells

    Directory of Open Access Journals (Sweden)

    Paul de Vos

    2017-08-01

    Full Text Available Orally ingested bacteria interact with intestinal mucosa and may impact immunity. However, insights in mechanisms involved are limited. In this randomized placebo-controlled cross-over trial, healthy human subjects were given Lactobacillus plantarum supplementation (strain TIFN101, CIP104448, or WCFS1 or placebo for 7 days. To determine whether L. plantarum can enhance immune response, we compared the effects of three stains on systemic and gut mucosal immunity, by among others assessing memory responses against tetanus toxoid (TT-antigen, and mucosal gene transcription, in human volunteers during induction of mild immune stressor in the intestine, by giving a commonly used enteropathic drug, indomethacin [non-steroidal anti-inflammatory drug (NSAID]. Systemic effects of the interventions were studies in peripheral blood samples. NSAID was found to induce a reduction in serum CD4+/Foxp3 regulatory cells, which was prevented by L. plantarum TIFN101. T-cell polarization experiments showed L. plantarum TIFN101 to enhance responses against TT-antigen, which indicates stimulation of memory responses by this strain. Cell extracts of the specific L. plantarum strains provoked responses after WCFS1 and TIFN101 consumption, indicating stimulation of immune responses against the specific bacteria. Mucosal immunomodulatory effects were studied in duodenal biopsies. In small intestinal mucosa, TIFN101 upregulated genes associated with maintenance of T- and B-cell function and antigen presentation. Furthermore, L. plantarum TIFN101 and WCFS1 downregulated immunological pathways involved in antigen presentation and shared downregulation of snoRNAs, which may suggest cellular destabilization, but may also be an indicator of tissue repair. Full sequencing of the L. plantarum strains revealed possible gene clusters that might be responsible for the differential biological effects of the bacteria on host immunity. In conclusion, the impact of oral consumption L

  15. PG-2, a Potent AMP against Pathogenic Microbial Strains, from Potato (Solanum tuberosum L cv. Gogu Valley Tubers Not Cytotoxic against Human Cells

    Directory of Open Access Journals (Sweden)

    Yoonkyung Park

    2013-02-01

    Full Text Available In an earlier study, we isolated potamin-1 (PT-1, a 5.6-kDa trypsin-chymotrypsin protease inhibitor, from the tubers of a potato strain (Solanum tuberosum L cv. Gogu Valley. We established that PT-1 strongly inhibits pathogenic microbial strains, but not human bacterial strains, and that its sequence shows 62% homology with a serine protease inhibitor. In the present study, we isolated an antifungal and antibacterial peptide with no cytotoxicity from tubers of the same potato strain. The peptide (peptide-G2, PG-2 was isolated using salt-extraction, ultrafiltration and reverse-phase high performance liquid chromatography (RP-HPLC. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS showed the protein to have a molecular mass of 3228.5 Da, while automated Edman degradation showed the N-terminal sequence of PG-2 to be LVKDNPLDISPKQVQALCTDLVIRCMCCC-. PG-2 exhibited antimicrobial activity against Candida albicans, a human pathogenic yeast strain, and Clavibacter michiganensis subsp. michiganensis, a plant late blight strain. PG-2 also showed antibacterial activity against Staphylococcus aureus, but did not lyse human red blood cells and was thermostable. Overall, these results suggest PG-2 may be a good candidate to serve as a natural antimicrobial agent, agricultural pesticide and/or food additive.

  16. PG-2, a Potent AMP against Pathogenic Microbial Strains, from Potato (Solanum tuberosum L cv. Gogu Valley) Tubers Not Cytotoxic against Human Cells

    Science.gov (United States)

    Kim, Jin-Young; Gopal, Ramamourthy; Kim, Sang Young; Seo, Chang Ho; Lee, Hyang Burm; Cheong, Hyeonsook; Park, Yoonkyung

    2013-01-01

    In an earlier study, we isolated potamin-1 (PT-1), a 5.6-kDa trypsin-chymotrypsin protease inhibitor, from the tubers of a potato strain (Solanum tuberosum L cv. Gogu Valley). We established that PT-1 strongly inhibits pathogenic microbial strains, but not human bacterial strains, and that its sequence shows 62% homology with a serine protease inhibitor. In the present study, we isolated an antifungal and antibacterial peptide with no cytotoxicity from tubers of the same potato strain. The peptide (peptide-G2, PG-2) was isolated using salt-extraction, ultrafiltration and reverse-phase high performance liquid chromatography (RP-HPLC). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) showed the protein to have a molecular mass of 3228.5 Da, while automated Edman degradation showed the N-terminal sequence of PG-2 to be LVKDNPLDISPKQVQALCTDLVIRCMCCC-. PG-2 exhibited antimicrobial activity against Candida albicans, a human pathogenic yeast strain, and Clavibacter michiganensis subsp. michiganensis, a plant late blight strain. PG-2 also showed antibacterial activity against Staphylococcus aureus, but did not lyse human red blood cells and was thermostable. Overall, these results suggest PG-2 may be a good candidate to serve as a natural antimicrobial agent, agricultural pesticide and/or food additive. PMID:23429275

  17. Evaluation of the safety of ancient strains of wheat in coeliac disease reveals heterogeneous small intestinal T cell responses suggestive of coeliac toxicity.

    Science.gov (United States)

    Šuligoj, Tanja; Gregorini, Armando; Colomba, Mariastella; Ellis, H Julia; Ciclitira, Paul J

    2013-12-01

    Coeliac disease is a chronic small intestinal immune-mediated enteropathy triggered by dietary gluten in genetically predisposed individuals. Since it is unknown if all wheat varieties are equally toxic to coeliac patients seven Triticum accessions showing different origin (ancient/modern) and ploidy (di-, tetra- hexaploid) were studied. Selected strains of wheat were ancient Triticum monococcum precoce (AA genome) and Triticum speltoides (BB genome), accessions of Triticum turgidum durum (AABB genome) including two ancient (Graziella Ra and Kamut) and two modern (Senatore Cappelli and Svevo) durum strains of wheat and Triticum aestivum compactum (AABBDD genome). Small intestinal gluten-specific T-cell lines generated from 13 coeliac patients were tested with wheat accessions by proliferation assays. All strains of wheat independent of ploidy or ancient/modern origin triggered heterogeneous responses covering wide ranges of stimulation indices. Ancient strains of wheat, although previously suggested to be low or devoid of coeliac toxicity, should be tested for immunogenicity using gluten-specific T-cell lines from multiple coeliac patients rather than gluten-specific clones to assess their potential toxicity. Our findings provide further evidence for the need for a strict gluten-free diet in coeliac patients, including avoidance of ancient strains of wheat. Copyright © 2013 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

  18. Strain variation among Bordetella pertussis isolates in finland, where the whole-cell pertussis vaccine has been used for 50 years.

    Science.gov (United States)

    Elomaa, Annika; Advani, Abdolreza; Donnelly, Declan; Antila, Mia; Mertsola, Jussi; Hallander, Hans; He, Qiushui

    2005-08-01

    Pertussis is an infectious disease of the respiratory tract caused by Bordetella pertussis. Despite the introduction of mass vaccination against pertussis in Finland in 1952, pertussis has remained an endemic disease with regular epidemics. To monitor changes in the Finnish B. pertussis population, 101 isolates selected from 1991 to 2003 and 21 isolates selected from 1953 to 1982 were studied together with two Finnish vaccine strains. The analyses included serotyping of fimbriae (Fim), genotyping of the pertussis toxin S1 subunit (ptxA) and pertactin (prn), and pulsed-field gel electrophoresis (PFGE) after digestion of B. pertussis genomic DNA with XbaI restriction enzyme. Strains isolated before 1977 were found to harbor the same ptxA as the strains used in the Finnish whole-cell pertussis vaccine, and strains isolated before 1982 harbored the same prn as the strains used in the Finnish whole-cell pertussis vaccine. All recent isolates, however, represented genotypes distinct from those of the two vaccine strains. A marked shift of predominant serotype from Fim serotype 2 (Fim2) to Fim3 has been observed since the late 1990s. Temporal changes were seen in the genome of B. pertussis by PFGE analysis. Three PFGE profiles (BpSR1, BpSR11, and BpSR147) were distinguished by their prevalence between 1991 and 2003. The yearly emergence of the three profiles was distributed periodically. Our study stresses the importance of the continuous monitoring of emerging strains of B. pertussis and the need to obtain a better understanding of the relationship of the evolution of B. pertussis in vaccinated populations.

  19. Strain Variation among Bordetella pertussis Isolates in Finland, Where the Whole-Cell Pertussis Vaccine Has Been Used for 50 Years

    Science.gov (United States)

    Elomaa, Annika; Advani, Abdolreza; Donnelly, Declan; Antila, Mia; Mertsola, Jussi; Hallander, Hans; He, Qiushui

    2005-01-01

    Pertussis is an infectious disease of the respiratory tract caused by Bordetella pertussis. Despite the introduction of mass vaccination against pertussis in Finland in 1952, pertussis has remained an endemic disease with regular epidemics. To monitor changes in the Finnish B. pertussis population, 101 isolates selected from 1991 to 2003 and 21 isolates selected from 1953 to 1982 were studied together with two Finnish vaccine strains. The analyses included serotyping of fimbriae (Fim), genotyping of the pertussis toxin S1 subunit (ptxA) and pertactin (prn), and pulsed-field gel electrophoresis (PFGE) after digestion of B. pertussis genomic DNA with XbaI restriction enzyme. Strains isolated before 1977 were found to harbor the same ptxA as the strains used in the Finnish whole-cell pertussis vaccine, and strains isolated before 1982 harbored the same prn as the strains used in the Finnish whole-cell pertussis vaccine. All recent isolates, however, represented genotypes distinct from those of the two vaccine strains. A marked shift of predominant serotype from Fim serotype 2 (Fim2) to Fim3 has been observed since the late 1990s. Temporal changes were seen in the genome of B. pertussis by PFGE analysis. Three PFGE profiles (BpSR1, BpSR11, and BpSR147) were distinguished by their prevalence between 1991 and 2003. The yearly emergence of the three profiles was distributed periodically. Our study stresses the importance of the continuous monitoring of emerging strains of B. pertussis and the need to obtain a better understanding of the relationship of the evolution of B. pertussis in vaccinated populations. PMID:16081896

  20. X-ray sensitivity of fifty-three human diploid fibroblast cell strains from patients with characterized genetic disorders

    Energy Technology Data Exchange (ETDEWEB)

    Weichselbaum, R.R.; Nove, J.; Little, J.B.

    1980-03-01

    The in vitro response of 53 human diploid fibroblast strains to x-irradiation was studied using a clonogenic survival assay. The strains, derived from patients with a variety of characterized clinical conditions, most with a genetic component, ranged in Do (a measure of the slope of the survival curve) from 43 to 168 rads. The mean Do's of six strains from normal individuals was 140 to 152 rads, with an overall range, based on the extremes of their standard errors, of 128 to 164 rads. Three-quarters of the strains studied fell within this range. Strains identified as sensitive came from patients with ataxia telangiectasia, progeria, the two genetic forms of retinoblastoma, and partial trisomy of chromosome 13. No marked radiosensitivity was found among strains derived from patients with a number of other conditions associated with a predisposition to malignancy.

  1. Highly pathogenic H5N1 influenza A virus strains provoke heterogeneous IFN-α/β responses that distinctively affect viral propagation in human cells.

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    Markus Matthaei

    Full Text Available The fatal transmissions of highly pathogenic avian influenza A viruses (IAV of the H5N1 subtype to humans and high titer replication in the respiratory tract indicate that these pathogens can overcome the bird-to-human species barrier. While type I interferons (IFN-α/β are well described to contribute to the species barrier of many zoonotic viruses, current data to the role of these antiviral cytokines during human H5N1 IAV infections is limited and contradictory. We hypothesized an important role for the IFN system in limiting productive infection of avian H5N1 strains in human cells. Hence, we examined IFN-α/β gene activation by different avian and human H5N1 isolates, if the IFN-α/β response restricts H5N1 growth and whether the different strains were equally capable to regulate the IFN-α/β system via their IFN-antagonistic NS1 proteins. Two human H5N1 isolates and a seasonal H3N2 strain propagated efficiently in human respiratory cells and induced little IFN-β, whereas three purely avian H5N1 strains were attenuated for replication and provoked higher IFN secretion. Replication of avian viruses was significantly enhanced on interferon-deficient cells, and exogenous IFN potently limited the growth of all strains in human cells. Moreover, IFN-α/β activation by all strains depended on retinoic acid-inducible gene I excluding principal differences in receptor activation between the different viruses. Interestingly, all H5N1 NS1 proteins suppressed IFN-α/β induction comparably well to the NS1 of seasonal IAV. Thus, our study shows that H5N1 strains are heterogeneous in their capacity to activate human cells in an NS1-independent manner. Our findings also suggest that H5N1 viruses need to acquire adaptive changes to circumvent strong IFN-α/β activation in human host cells. Since no single amino acid polymorphism could be associated with a respective high- or low induction phenotype we propose that the necessary adaptations to

  2. Effect of environmental factors and cell physiological state on Pulsed Electric Fields resistance and repair capacity of various strains of Escherichia coli.

    Science.gov (United States)

    Somolinos, M; García, D; Mañas, P; Condón, S; Pagán, R

    2008-06-10

    The aim was to determine the resistance variation of four strains of Escherichia coli to Pulsed Electric Fields (PEF), the role of the sigma factor RpoS in PEF resistance, as well as the influence of several environmental factors and the cell physiological state on the PEF resistance and repair capacity. The rpoS null mutant, E. coli BJ4L1, exhibited decreased PEF resistance as compared with its wild-type parent, BJ4. W3110 and O157:H7 were the most PEF-resistant strains: whereas 2 and more than 3 Log10 cycles of BJ4 and BJ4L1 cells, respectively, were inactivated after 50 pulses at 35 kV/cm, only 0.5 Log10 cycle of inactivation of W3110 and O157:H7 was attained. A different pattern was observed and the resistance variation among strains was largely reduced, when selective recovery media were used. At exponential growth phase, the resistance of the four strains was lower, and more than 4 Log10 cycles of inactivation of all strains tested were attained at 30 kV/cm. Previous heat and cold shock treatments scarcely influenced cell PEF resistance. PEF survival increased with the reduction in water activity of the treatment medium to 0.94: the occurrence of sublethally injured cells was negligible, and less than 1 Log10 cycle of inactivation was attained at 35 kV/cm. PEF-treated cells were sensitive to a subsequent storage at pH 4.0 or in the presence of sorbic acid, attaining a final inactivation of 4-5 Log10 cycles after 24 hour-incubation. In conclusion, the work confirms the role of rpoS in PEF resistance. E. coli strains exhibit large differences in PEF resistance. These differences were less important when cells were recovered under selective conditions. Both resistance variation among strains and occurrence of sublethal damage were noticeably influenced by the environmental factors tested.

  3. Avian influenza A virus PB2 promotes interferon type I inducing properties of a swine strain in porcine dendritic cells

    Energy Technology Data Exchange (ETDEWEB)

    Ocana-Macchi, Manuela; Ricklin, Meret E.; Python, Sylvie; Monika, Gsell-Albert [Institute of Virology and Immunoprophylaxis, Mittelhaeusern (Switzerland); Stech, Juergen; Stech, Olga [Friedrich-Loeffler Institut, Greifswald-Insel Riems (Germany); Summerfield, Artur, E-mail: artur.summerfield@ivi.admin.ch [Institute of Virology and Immunoprophylaxis, Mittelhaeusern (Switzerland)

    2012-05-25

    The 2009 influenza A virus (IAV) pandemic resulted from reassortment of avian, human and swine strains probably in pigs. To elucidate the role of viral genes in host adaptation regarding innate immune responses, we focussed on the effect of genes from an avian H5N1 and a porcine H1N1 IAV on infectivity and activation of porcine GM-CSF-induced dendritic cells (DC). The highest interferon type I responses were achieved by the porcine virus reassortant containing the avian polymerase gene PB2. This finding was not due to differential tropism since all viruses infected DC equally. All viruses equally induced MHC class II, but porcine H1N1 expressing the avian viral PB2 induced more prominent nuclear NF-{kappa}B translocation compared to its parent IAV. The enhanced activation of DC may be detrimental or beneficial. An over-stimulation of innate responses could result in either pronounced tissue damage or increased resistance against IAV reassortants carrying avian PB2.

  4. Efficient infectious cell culture systems of the hepatitis C virus (HCV) prototype strains HCV-1 and H77

    DEFF Research Database (Denmark)

    Li, Yi-Ping; Ramirez, Santseharay; Mikkelsen, Lotte

    2015-01-01

    UNLABELLED: The first discovered and sequenced hepatitis C virus (HCV) genome and the first in vivo infectious HCV clones originated from the HCV prototype strains HCV-1 and H77, respectively, both widely used in research of this important human pathogen. In the present study, we developed...... mutations (HCV1cc) replicated efficiently in Huh7.5 cells and produced supernatant infectivity titers of 10(4.0) focus-forming units (FFU)/ml. Eight of these mutations were identified from passaged HCV-1 viruses, and the A970T/I1312V/C2419R/A2919T mutations were essential for infectious particle production......-adapted full-length TN viruses and a common NS3 helicase mutation (S1368P) derived from viable H77C and HCV-1 5-5A recombinants, initiated replication and culture adaptation of H77C containing LSG and TNcc(1a)-adaptive mutations. An H77C recombinant harboring 19 mutations (H77Ccc) replicated and spread...

  5. Nucleotide sequencing of hepatitis A virus Js-4 strain subcultured in human diploid cells%甲型肝炎病毒Js-4株经人二倍体细胞传代后的核苷酸序列分析

    Institute of Scientific and Technical Information of China (English)

    胡艳灵; 陈统球; 黄明; 戚凤春; 崔燕萍; 潘翔

    2012-01-01

    目的 分析甲型肝炎病毒(Hepatitis A virus,HAV)Js-4株经人二倍体细胞MRC-5传代后的核苷酸序列变异情况.方法 将HAV Js-4株经Vero细胞适应14代获得的原始株Js-4-V14经人二倍体细胞MRC-5于35℃连续适应培养15代,取其中8个适应株(Js-4-M2、Js-4-M3、Js-4-M4、Js-4-M5、Js-4-M10、Js-4-M12、Js-4-M14、Js-4-M15)及原始株,提取病毒基因组RNA,通过RTPCR扩增病毒全基因组,采用生物信息学软件DNAstar Lasergene 7进行序列分析.结果 8个适应株全基因组与Js-4-V14株的核苷酸序列同源性在99.8%~100.0%之间;第5代适应株在5′-NCR(101~ 200 bp)发生较大变异,表现为105~157bp缺失,之后该突变稳定遗传;第10、12、14、15代适应株在2A(3 012 ~3 210 bp)区段3 122位点发生点突变;8个适应株和Js-4-V14毒株与野毒株HM175为同一基因亚型,均为IB型;9个毒株的主要抗原位点与野生株HM175完全一致.结论 HAV Js-4-V14株在MRC-5细胞传代适应过程中,主要抗原位点未发生突变,其与细胞适应性和病毒增殖有关的基因在传代早期已经变异,传代后期这些变异保持相对稳定.%Objective To analyze the variation of nucleotide sequence of hepatitis A virus (HAV) Js-4 strain after subculture in human diploid MRC-5 cells. Methods HAV Js-4 strain was subcultured in Vero cells for 14 passages, and the obtained Js-4-V14 strain was inoculated to MRC-5 cells and further subcultured at 35 Xl for 15 passages. Genomic RNAs were extracted from Js-4-V14 strain as well as eight adapted strains, i. e. Js-4-M2, Js-4-M3, Js-4-M4, Js-4-M5, Js-4-M10, Js-4-Mu, Js-4-M14 and Js-4-Mi5, with which the whole genomes were amplified by RT-PCR and sequenced by bioinformatics software DNAstar Lasergene 1. Results The homologies of whole genomes of eight adapted strains to those of strain Js-4-V14 were 99. 8% ~ 100. 0%. However, obvious variation was observed in 5'-NCR (101 ~ 200 bp) of strain Js-4-M5, expressed as deletion

  6. CagA-positive Helicobacter pylori strain containing three EPIYA C phosphorylation sites produces increase of G cell and decrease of D cell in experimentally infected gerbils (Meriones unguiculatus).

    Science.gov (United States)

    Júnior, Moacir Ferreira; Batista, Sérgio de Assis; Barbuto, Rafael Calvão; Gomes, Adriana Dias; Queiroz, Dulciene Maria Magalhães; Araújo, Ivana Duval; Caliari, Marcelo Vidigal

    2016-09-01

    Human infection by Helicobacter pylori is associated with an increase in the number of gastrin-producing G cells and a concomitant decrease of somatostatin-producing D cells. However, to our knowledge, changes in G and D cell numbers in response to infection with H. pylori CagA-positive strains containing different number of EPIYA-C phosphorylation sites have not been analyzed to date. Therefore, the aim of this study was to perform a quantitative analysis of the number of G and D cells in Mongolian gerbils challenged with H. pylori strains with different numbers of EPIYA-C motifs. Mongolian gerbils were inoculated with isogenic H. pylori strains containing one to three phosphorylation sites. Mucosal fragments were evaluated by morphometry and immunohistochemistry using primary polyclonal rabbit anti-gastrin and anti-somatostatin antibodies. Positive cells were counted using an image analyzer. Forty-five days after infection, there was a decrease in the number of D cells and an increase in the G/D cell ratio in the group with three EPIYA-C. Six months after infection, there was a progressive and significant increase in the number of G cells and in the G/D cell ratio, with a concomitant decrease in the number of D cells, especially in the three EPIYA-C group. CagA-positive H. pylori strains containing a large number of EPIYA-C phosphorylation sites induce a decrease in D cell number and an increase in G cell number and G/D ratio, which were correlated with the number of inflammatory cells of the lamina propria. Copyright © 2016 Medical University of Bialystok. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  7. Brucella suis Vaccine Strain 2 Induces Endoplasmic Reticulum Stress that Affects Intracellular Replication in Goat Trophoblast Cells In vitro.

    Science.gov (United States)

    Wang, Xiangguo; Lin, Pengfei; Li, Yang; Xiang, Caixia; Yin, Yanlong; Chen, Zhi; Du, Yue; Zhou, Dong; Jin, Yaping; Wang, Aihua

    2016-01-01

    Brucella has been reported to impair placental trophoblasts, a cellular target where Brucella efficiently replicates in association with the endoplasmic reticulum (ER), and ultimately trigger abortion in pregnant animals. However, the precise effects of Brucella on trophoblast cells remain unclear. Here, we describe the infection and replication of Brucella suis vaccine strain 2 (B.suis.S2) in goat trophoblast cells (GTCs) and the cellular and molecular responses induced in vitro. Our studies demonstrated that B.suis.S2 was able to infect and proliferate to high titers, hamper the proliferation of GTCs and induce apoptosis due to ER stress. Tunicamycin (Tm), a pharmacological chaperone that strongly mounts ER stress-induced apoptosis, inhibited B.suis.S2 replication in GTCs. In addition, 4 phenyl butyric acid (4-PBA), a pharmacological chaperone that alleviates ER stress-induced apoptosis, significantly enhanced B.suis.S2 replication in GTCs. The Unfolded Protein Response (UPR) chaperone molecule GRP78 also promoted B.suis.S2 proliferation in GTCs by inhibiting ER stress-induced apoptosis. We also discovered that the IRE1 pathway, but not the PERK or ATF6 pathway, was activated in the process. However, decreasing the expression of phosphoIRE1α and IRE1α proteins with Irestatin 9389 (IRE1 antagonist) in GTCs did not affect the proliferation of B.suis.S2. Although GTC implantation was not affected upon B.suis.S2 infection, progesterone secretion was suppressed, and prolactin and estrogen secretion increased; these effects were accompanied by changes in the expression of genes encoding key steroidogenic enzymes. This study systematically explored the mechanisms of abortion in Brucella infection from the viewpoint of pathogen invasion, ER stress and reproductive endocrinology. Our findings may provide new insight for understanding the mechanisms involved in goat abortions caused by Brucella infection.

  8. Brucella suis vaccine strain 2 induces endoplasmic reticulum stress that affects intracellular replication in goat trophoblast cells in vitro

    Directory of Open Access Journals (Sweden)

    Xiangguo eWang

    2016-02-01

    Full Text Available Brucella has been reported to impair placental trophoblasts, a cellular target where Brucella efficiently replicates in association with the endoplasmic reticulum (ER, and ultimately trigger abortion in pregnant animals. However, the precise effects of Brucella on trophoblast cells remain unclear. Here, we describe the infection and replication of Brucella suis vaccine strain 2 (B.suis.S2 in goat trophoblast cells (GTCs and the cellular and molecular responses induced in vitro. Our studies demonstrated that B.suis.S2 was able to infect and proliferate to high titers, hamper the proliferation of GTCs and induce apoptosis due to ER stress. Tunicamycin (Tm, a pharmacological chaperone that strongly mounts ER stress-induced apoptosis, inhibited B.suis.S2 replication in GTCs. In addition, 4 phenyl butyric acid (4-PBA, a pharmacological chaperone that alleviates ER stress-induced apoptosis, significantly enhanced B.suis.S2 replication in GTCs. The Unfolded Protein Response (UPR chaperone molecule GRP78 also promoted B.suis.S2 proliferation in GTCs by inhibiting ER stress-induced apoptosis. We also discovered that the IRE1 pathway, but not the PERK or ATF6 pathway, was activated in the process. However, decreasing the expression of phosphoIRE1α and IRE1α proteins with Irestatin 9389 (IRE1 antagonist in GTCs did not affect the proliferation of B.suis.S2. Although GTC implantation was not affected upon B.suis.S2 infection, progesterone secretion was suppressed, and prolactin and estrogen secretion increased; these effects were accompanied by changes in the expression of genes encoding key steroidogenic enzymes. This study systematically explored the mechanisms of abortion in Brucella infection from the viewpoint of pathogen invasion, ER stress and reproductive endocrinology. Our findings may provide new insight for understanding the mechanisms involved in goat abortions caused by Brucella infection.

  9. Polymorphism of 41 kD Flagellin Gene and Its Human B-Cell Epitope in Borrelia burgdorferi Strains of China

    Directory of Open Access Journals (Sweden)

    Huixin Liu

    2016-01-01

    Full Text Available The 41 kD flagellin of Borrelia burgdorferi (B. burgdorferi is a major component of periplasmic flagellar filament core and a good candidate for serodiagnosis in early stage of Lyme disease. Here, we chose 89 B. burgdorferi strains in China, amplified the gene encoding the 41 kD flagellin, and compared the sequences. The results showed that genetic diversity presented in the 41 kD flagellin genes of all 89 strains among the four genotypes of B. burgdorferi, especially in the genotype of B. garinii. Some specific mutation sites for each genotype of the 41 kD flagellin genes were found, which could be used for genotyping B. burgdorferi strains in China. Human B-cell epitope analysis showed that thirteen of 15 nonsynonymous mutations occurred in the epitope region of 41 kD flagellin and thirty of 42 B-cell epitopes were altered due to all 13 nonsynonymous mutations in the epitope region, which may affect the function of the antigen. Nonsynonymous mutations and changed human B-cell epitopes exist in 41 kD flagellin of B. burgdorferi sensu lato strains; these changes should be considered in serodiagnosis of Lyme disease.

  10. Mycobacterium tuberculosis Multidrug-Resistant Strain M Induces Low IL-8 and Inhibits TNF-α Secretion by Bronchial Epithelial Cells Altering Neutrophil Effector Functions

    Directory of Open Access Journals (Sweden)

    Denise Kviatcovsky

    2017-01-01

    Full Text Available M strain, the most prevalent multidrug-resistant strain of Mycobacterium tuberculosis (Mtb in Argentina, has mounted mechanisms to evade innate immune response. The role of human bronchial epithelium in Mtb infection remains unknown as well as its crosstalk with neutrophils (PMN. In this work, we evaluate whether M and H37Rv strains invade and replicate within bronchial epithelial cell line Calu-6 and how conditioned media (CM derived from infected cells alter PMN responses. We demonstrated that M infects and survives within Calu-6 without promoting death. CM from M-infected Calu-6 (M-CM did not attract PMN in correlation with its low IL-8 content compared to H37Rv-CM. Also, PMN activation and ROS production in response to irradiated H37Rv were impaired after treatment with M-CM due to the lack of TNF-α. Interestingly, M-CM increased H37Rv replication in PMN which would allow the spreading of mycobacteria upon PMN death and sustain IL-8 release. Thus, our results indicate that even at low invasion/replication rate within Calu-6, M induces the secretion of factors altering the crosstalk between these nonphagocytic cells and PMN, representing an evasion mechanism developed by M strain to persist in the host. These data provide new insights on the role of bronchial epithelium upon M infection.

  11. Behaviour of Saccharomyces cerevisiae wine strains during adaptation to unfavourable conditions of fermentation on synthetic medium: cell lipid composition, membrane integrity, viability and fermentative activity.

    Science.gov (United States)

    Mannazzu, Ilaria; Angelozzi, Daniele; Belviso, Simona; Budroni, Marilena; Farris, Giovanni Antonio; Goffrini, Paola; Lodi, Tiziana; Marzona, Mario; Bardi, Laura

    2008-01-15

    During must fermentation wine strains are exposed to a variety of biotic and abiotic stresses which, when prevailing over the cellular defence systems, can affect cell viability with negative consequences on the progression of the fermentative process. To investigate the ability of wine strains to survive and adapt to unfavourable conditions of fermentation, the lipid composition, membrane integrity, cell viability and fermentative activity of three strains of Saccharomyces cerevisiae were analysed during hypoxic growth in a sugar-rich medium lacking lipid nutrients. These are stressful conditions, not unusual during must fermentation, which, by affecting lipid biosynthesis may exert a negative effect on yeast viability. The results obtained showed that the three strains were able to modulate cell lipid composition during fermentation. However, only two of them, which showed highest viability and membrane integrity at the end of the fermentation process, reached a fatty acid composition which seemed to be optimal for a successful adaptation. In particular, C16/TFA and UFA/TFA ratios, more than total lipid and ergosterol contents, seem to be involved in yeast adaptation.

  12. Influence of the impoundment of the Three Gorges Reservoir on the micro-seismicity and the 2013 M5.1 Badong earthquake (Yangtze, China)

    Science.gov (United States)

    Zhang, Huai; Cheng, Huihong; Pang, Yajin; Shi, Yaolin; Yuen, David A.

    2016-12-01

    On December 16, 2013, right after the Three Gorges Reservoir (TGR) reached its highest annual water level, a powerful M5.1 earthquake occurred in Badong County, China's Hubei Province. The epicenter is 5.5 km away from the upstream boundary and 100 km from the dam. Was this earthquake triggered by the impoundment of the TGR, and what are its subsequences? To answer these questions, we constructed a coupled three-dimensional poroelastic finite element model to examine the ground surface deformation, the Coulomb failure stress change (ΔCFS) due to the variation of elastic stress and pore pressure, and the elastic strain energy potential accumulation in the TGR region upon the occurrence of this event. Our calculated maximum surface deformation values beneath the TGR compare well with GPS observations, which validates our numerical model. At the hypocenter of the earthquake, ΔCFS is around 8.0 ∼ 11.0 kPa, revealing that it may be eventually triggered by the impoundment. We also discovered that the total elastic strain energy potential accumulation due to the impounded water load is around 1.7 × 1012 J, merely equivalent to 0.01% of the total energy released by this event, indicating that this earthquake is predominately controlled by the typical regional tectonic settings as well as the weak fault zones, and the reservoir impoundment might only facilitate its procedure or occurrence. Furthermore, the stress level in this region remains high after this earthquake and the subsequent reservoir-triggered micro-seismicity or even bigger event are highly possible.

  13. DNMT3A R882 mutation is associated with elevated expression of MAFB and M4/M5 immunophenotype of acute myeloid leukemia blasts.

    Science.gov (United States)

    Yang, Li; Liu, Ya'Nan; Zhu, Li; Xiao, Min

    2015-01-01

    Researchers have recognized that aberrant methylation is an important initiating event in the pathogenesis of hematological malignancies. DNMT3A is a DNA methyltransferase that plays a vital role in de novo methylation of DNA. Somatic mutation of DNMT3A, especially at the Arg882 (R882) site of the DNMT3A coding sequence, has been identified in pre-leukemic stem cell clones as one of the driver mutations of acute myeloid leukemia (AML). Statistical analysis has indicated that patients with AML with DNMT3A mutation tend to have the M4/M5 subtype of AML according to the French-American-British classification. In this study we aimed to investigate the association between the typical immunophenotype of leukemic blasts and mutation of DNMT3A R882. In addition, we further determined the relationship between DNMT3A R882 mutation and the expression of monocytic differentiation genes, and its clinical significance.

  14. Histomorphometric characteristics of immune cells in small intestine of pigs perorally immunized with vaccine candidate F18ac+ nonenterotoxigenic E. coli strain

    Directory of Open Access Journals (Sweden)

    I. Valpotic

    2009-12-01

    Full Text Available Colidiarrhea and colienterotoxemia caused by F4+ and/or F18+ enterotoxigenic E. coli (ETEC strains are the most prevalent infections of suckling and weaned pigs. Here we tested the immunogenicity and protective effectiveness of attenuated F18ac+ non-ETEC vaccine candidate strain against challenge infection with F4ac+ ETEC strain by quantitative phenotypic analysis of small intestinal leukocyte subsets in weaned pigs. We also evaluated levamisole as an immune response modifier (IRM and its adjuvanticity when given in the combination with the experimental vaccine. The pigs were parenterally immunized with either levamisole (at days -2, -1 and 0 or with levamisole and perorally given F18ac+ non-ETEC strain (at day 0, and challenged with F4ac+ ETEC strain 7 days later. At day 13 the pigs were euthanatized and sampled for immunohistological/histomorphometrical analyses. Lymphoid CD3+, CD45RA+, CD45RC+, CD21+, IgA+ and myeloid SWC3+ cell subsets were identified in jejunal and ileal epithelium, lamina propria and Peyer’s patches using the avidin-biotin complex method, and their numbers were determined by computer-assisted histomorphometry. Quantitative immunophenotypic analyses showed that levamisole treated pigs had highly increased numbers of jejunal CD3+, CD45RC+ and SWC3+ cells (p<0.05 as compared to those recorded in nontreated control pigs. In the ileum of these pigs we have recorded that only CD21+ cells were significantly increased (p<0.01. The pigs that were treated with levamisole adjuvanted experimental vaccine had significantly increased numbers of all tested cell subsets in both segments of the small intestine. It was concluded that levamisole adjuvanted F18ac+ non-ETEC vaccine was a requirement for the elicitation of protective gut immunity in this model; nonspecific immunization with levamisole was less effective, but confirmed its potential as an IRM.

  15. Diseño del mecanismo de rotación del espejo M5 de un telescopio reflector

    OpenAIRE

    Fernández Gómez, Jorge

    2010-01-01

    El objetivo principal de este proyecto fin de carrera consiste en diseñar una solución para el desplazamiento de uno de los espejos, denominado M5 (Mirror 5), que forman el sistema óptico del telescopio E-ELT. Para mover dicho espejo se hace necesario desarrollar un mecanismo que le permita rotar 180º alrededor del eje de azimut desde una posición A, hasta una posición B, según apunte a una estación focal o a otra. El mecanismo debe de ser capaz de realizar la maniobra con rapidez, seguridad,...

  16. Cell wall ultrastructure of flocculent and non-flocculent Schizosaccharomyces pombe strains. Effect of cell wall hydrolysing enzymes on flocculation and cell wall ultastructure.

    Science.gov (United States)

    Geleta, Anna; Kristóf, Z; Maráz, Anna

    2007-03-01

    Scanning and transmission electron microscopic studies revealed the presence of slime-like, amorphous material on the surface of Schizosaccahromyces pombe RIVE 4-2-1 cells, independently, whether they were in flocculated or in non-flocculated state. Close contact of the adjacent cells via the merging outermost cell wall layers was found, however, only in the case of floc formation, which was induced by cultivating the cells in the presence of 6% (v/v) ethanol. Irreversible loss of the flocculation ability of the cells by treatment with proteinases suggests that proteinaceous cell surface molecules as lectins contribute to the cell-to-cell interaction during flocculation. Both proteinase K and pronase treatments removed a distinct outer layer of the cell wall, which indicated that the protein moieties of the phosphogalactomannan outer surface layer has a crucial role in the maintenance of cell wall integrity. In the case of lysing enzyme treatment the removal of the outermost layer was also observed as the first step of the cell wall digestion, while driselase treatment resulted in almost complete digestion of the cell wall.

  17. Extent of Systemic Spread Determines CD8+ T Cell Immunodominance for Laboratory Strains, Smallpox Vaccines, and Zoonotic Isolates of Vaccinia Virus.

    Science.gov (United States)

    Flesch, Inge E A; Hollett, Natasha A; Wong, Yik Chun; Quinan, Bárbara Resende; Howard, Debbie; da Fonseca, Flávio G; Tscharke, David C

    2015-09-01

    CD8(+) T cells that recognize virus-derived peptides presented on MHC class I are vital antiviral effectors. Such peptides presented by any given virus vary greatly in immunogenicity, allowing them to be ranked in an immunodominance hierarchy. However, the full range of parameters that determine immunodominance and the underlying mechanisms remain unknown. In this study, we show across a range of vaccinia virus strains, including the current clonal smallpox vaccine, that the ability of a strain to spread systemically correlated with reduced immunodominance. Reduction in immunodominance was observed both in the lymphoid system and at the primary site of infection. Mechanistically, reduced immunodominance was associated with more robust priming and especially priming in the spleen. Finally, we show this is not just a property of vaccine and laboratory strains of virus, because an association between virulence and immunodominance was also observed in isolates from an outbreak of zoonotic vaccinia virus that occurred in Brazil.

  18. Detection of weak estrogenic flavonoids using a recombinant yeast strain and a modified MCF7 cell proliferation assay.

    Science.gov (United States)

    Breinholt, V; Larsen, J C

    1998-06-01

    A newly developed recombinant yeast strain, in which the human estrogen receptor has been stably integrated into the genome of the yeast, was used to gain information on the estrogenic activity of a large series of dietary flavonoids. Among 23 flavonoids investigated, 8 were found to markedly stimulate the transcriptional activity of the human estrogen receptor in the yeast assay increasing transcriptional activity 5-13-fold above background level, corresponding to EC50 values between 0.1 and 25 microM. Five compounds increased the transcriptional activity 2-5-fold over the control, with EC50 values ranging from 84 to 102 microM, whereas the remaining flavonoids were devoid of activity. The most potent flavonoid estrogens tested were naringenin, apigenin, kaempferol, phloretin, and the four isoflavonoids equol, genistein, daidzein, and biochanin A. With the exception of biochanin A, the main feature required to confer estrogenicity was the presence of a single hydroxyl group in the 4'-position of the B-ring of the flavan nucleus, corresponding to the 4-position on phloretin. The estrogenic potency of the flavonoids was found to be 4 000-4 000 000 times lower than that observed for 17beta-estradiol, when compared on the basis of EC50 values. The estrogenic activity of the dietary flavonoids was further investigated in estrogen-dependent human MCF7 breast cancer cells. In this system several of the flavonoids were likewise capable of mimicking natural estrogens and thereby induce cell proliferation. Similar structural requirements for estrogenic activity were found for the two assays. The present results provide evidence that several of the flavo-estrogens possess estrogenic properties comparable in activity to the well-established isoflavonoid estrogens. The use of Alamar Blue, a vital dye which is metabolically reduced by cellular enzymes to a fluorescent product, was found to greatly simplify the MCF7 cell-based estrogen screen, making this mammalian assay

  19. Effects of cyclic longitudinal mechanical strain and dexamethasone on osteogenic differentiation of human bone marrow stromal cells

    Directory of Open Access Journals (Sweden)

    Jagodzinski M.

    2004-04-01

    Full Text Available The aim of the study was to investigate the effect of cyclic mechanical strain on differentiation markers in the presence or absence of dexamethasone. Human bone marrow stromal cells (BMSC from seven donors (32.5±6.2 years were cultivated with (D+ or without (D- dexamethasone. A cyclic mechanical strain with an elongation of 2% (D+2; D-2 or 8% (D+8; D-8 was applied for three days with a stimulation time of three times two hours each day. Levels of alkaline phosphatase (ALP and osteocalcin (OC were compared after time intervals of four and seven days. mRNA expression of Collagen I, III and Cbfa1 was investigated after one, four, and seven days. ALP levels were significantly increased in the D+8 group after four and seven days (147.1±6.3%; p<0.05 and 168.6±6,5%; p<0.03 and in the D-8 group after 7 days (197.4±10.4; p<0.04. Cyclic strain had a significant influence on ALP-secretion (F=7.5; p<0.01. In the D-8 group there was a significant increase in OC secretion after 4 days (140.9±12.5%; p<0.05.; p<0.01. The effect of stretching was significantly stronger than that of dexamethasone (F=17.2 vs. 1.8. Collagen I (Col I expression was upregulated in D+8 cultures after 4 days (215.0±53.3 p<0.04 and after seven days (166.7±55.7; p<0.04. Collagen III (Col III expression was upregulated in D+2 and D+8 cultures after 4 days (200.7±16.3 and 185.9±12.7; p<0.04 and after seven days (154.4±10.1 and 118.8±16.4; p<0.04. There was a significant increase of Cbfa1 expression in D+8 cultures at all investigated time intervals (day 1: 105.5±3.7%; day 4: 104.7±3.0%; day 7: 104.4±2.1%; p<0.03. Stretching (F=20.0; p<0.01 was a stronger contributor to Cbfa-1 expression than dexamethasone (F=12.1; p<0.01. Cyclical mechanical stimulation with 8% elongation increases ALP and OC levels and upregulates Col I and III synthesis and Cbfa1 expression. In the short term, cyclical stretching is a stronger differentiation factor than dexamethasone. Cyclical stretching

  20. Modulation of intestinal inflammation by yeasts and cell wall extracts: strain dependence and unexpected anti-inflammatory role of glucan fractions.

    Directory of Open Access Journals (Sweden)

    Samir Jawhara

    Full Text Available Yeasts and their glycan components can have a beneficial or adverse effect on intestinal inflammation. Previous research has shown that the presence of Saccharomyces cerevisiae var. boulardii (Sb reduces intestinal inflammation and colonization by Candida albicans. The aim of this study was to identify dietary yeasts, which have comparable effects to the anti-C. albicans and anti-inflammatory properties of Sb and to assess the capabilities of yeast cell wall components to modulate intestinal inflammation. Mice received a single oral challenge of C. albicans and were then given 1.5% dextran-sulphate-sodium (DSS for 2 weeks followed by a 3-day restitution period. S. cerevisiae strains (Sb, Sc1 to Sc4, as well as mannoprotein (MP and β-glucan crude fractions prepared from Sc2 and highly purified β-glucans prepared from C. albicans were used in this curative model, starting 3 days after C. albicans challenge. Mice were assessed for the clinical, histological and inflammatory responses related to DSS administration. Strain Sc1-1 gave the same level of protection against C. albicans as Sb when assessed by mortality, clinical scores, colonization levels, reduction of TNFα and increase in IL-10 transcription. When Sc1-1 was compared with the other S. cerevisiae strains, the preparation process had a strong influence on biological activity. Interestingly, some S. cerevisiae strains dramatically increased mortality and clinical scores. Strain Sc4 and MP fraction favoured C. albicans colonization and inflammation, whereas β-glucan fraction was protective against both. Surprisingly, purified β-glucans from C. albicans had the same protective effect. Thus, some yeasts appear to be strong modulators of intestinal inflammation. These effects are dependent on the strain, species, preparation process and cell wall fraction. It was striking that β-glucan fractions or pure β-glucans from C. albicans displayed the most potent anti-inflammatory effect in the

  1. Modulation of intestinal inflammation by yeasts and cell wall extracts: strain dependence and unexpected anti-inflammatory role of glucan fractions.

    Science.gov (United States)

    Jawhara, Samir; Habib, Khalid; Maggiotto, François; Pignede, Georges; Vandekerckove, Pascal; Maes, Emmanuel; Dubuquoy, Laurent; Fontaine, Thierry; Guerardel, Yann; Poulain, Daniel

    2012-01-01

    Yeasts and their glycan components can have a beneficial or adverse effect on intestinal inflammation. Previous research has shown that the presence of Saccharomyces cerevisiae var. boulardii (Sb) reduces intestinal inflammation and colonization by Candida albicans. The aim of this study was to identify dietary yeasts, which have comparable effects to the anti-C. albicans and anti-inflammatory properties of Sb and to assess the capabilities of yeast cell wall components to modulate intestinal inflammation. Mice received a single oral challenge of C. albicans and were then given 1.5% dextran-sulphate-sodium (DSS) for 2 weeks followed by a 3-day restitution period. S. cerevisiae strains (Sb, Sc1 to Sc4), as well as mannoprotein (MP) and β-glucan crude fractions prepared from Sc2 and highly purified β-glucans prepared from C. albicans were used in this curative model, starting 3 days after C. albicans challenge. Mice were assessed for the clinical, histological and inflammatory responses related to DSS administration. Strain Sc1-1 gave the same level of protection against C. albicans as Sb when assessed by mortality, clinical scores, colonization levels, reduction of TNFα and increase in IL-10 transcription. When Sc1-1 was compared with the other S. cerevisiae strains, the preparation process had a strong influence on biological activity. Interestingly, some S. cerevisiae strains dramatically increased mortality and clinical scores. Strain Sc4 and MP fraction favoured C. albicans colonization and inflammation, whereas β-glucan fraction was protective against both. Surprisingly, purified β-glucans from C. albicans had the same protective effect. Thus, some yeasts appear to be strong modulators of intestinal inflammation. These effects are dependent on the strain, species, preparation process and cell wall fraction. It was striking that β-glucan fractions or pure β-glucans from C. albicans displayed the most potent anti-inflammatory effect in the DSS model.

  2. Experimental infection of Haemonchus contortus strains resistant and susceptible to benzimidazoles and the effect on mast cells distribution in the stomach of Mongolian gerbils (Meriones unguiculatus).

    Science.gov (United States)

    Königová, Alzbeta; Hrckova, Gabriela; Velebný, Samuel; Corba, Július; Várady, Marián

    2008-03-01

    Establishment rate of Haemonchus contortus in non-suppressed and immunosuppressed gerbils within 14 days post-infection was compared after inoculation with 1,000 third-stage larvae (L3), exsheathed BZ-susceptible larvae. Based on significantly higher number of larvae in gerbils receiving low doses of immunosuppressant agent hydrocortisone, development of benzimidazole (BZ)-susceptible and BZ-resistant strain of nematode in the stomach was studied on days 4, 7, 10, and 14 p.i. Sections of stomach from both groups of animals were examined for overall histopathological response and dynamics of mucosal mast cells (MMC) and connective tissue mast cells (CTMC). In the immunosuppressed gerbils, H. contortus L3 stage larvae developed to the L4 stage on days 10 and 14 p.i., and their sex ratio was higher toward female worms. Significantly higher ratios of establishment rate were recorded for BZ-susceptible than BZ-resistant strain. Infection elicited strong inflammation mainly in the lamina propria mucosae, where MMC numbers peaked on day 7 p.i., being present in a significantly higher numbers in gerbils infected with BZ-susceptible strain. Infection with BZ-susceptible strain of nematode also resulted in a higher number of CTMC in comparison with the effect of BZ-resistant strain, which were observed in the tela submucosa only. Thus, H. contortus infection in gerbils seems to be a suitable model to study host-parasite interactions. Our results indicate that BZ-resistant strain of H. contortus have a decreased capacity to establish infection in direct relation with lower mucosal and connective tissue MCs counts in the stomach.

  3. Genome-Wide Immune Modulation of TLR3-Mediated Inflammation in Intestinal Epithelial Cells Differs between Single and Multi-Strain Probiotic Combination

    Science.gov (United States)

    MacPherson, Chad W.; Shastri, Padmaja; Mathieu, Olivier; Tompkins, Thomas A.; Burguière, Pierre

    2017-01-01

    Genome-wide transcriptional analysis in intestinal epithelial cells (IEC) can aid in elucidating the impact of single versus multi-strain probiotic combinations on immunological and cellular mechanisms of action. In this study we used human expression microarray chips in an in vitro intestinal epithelial cell model to investigate the impact of three probiotic bacteria, Lactobacillus helveticus R0052 (Lh-R0052), Bifidobacterium longum subsp. infantis R0033 (Bl-R0033) and Bifidobacterium bifidum R0071 (Bb-R0071) individually and in combination, and of a surface-layer protein (SLP) purified from Lh-R0052, on HT-29 cells’ transcriptional profile to poly(I:C)-induced inflammation. Hierarchical heat map clustering, Set Distiller and String analyses revealed that the effects of Lh-R0052 and Bb-R0071 diverged from those of Bl-R0033 and Lh-R0052-SLP. It was evident from the global analyses with respect to the immune, cellular and homeostasis related pathways that the co-challenge with probiotic combination (PC) vastly differed in its effect from the single strains and Lh-R0052-SLP treatments. The multi-strain PC resulted in a greater reduction of modulated genes, found through functional connections between immune and cellular pathways. Cytokine and chemokine analyses based on specific outcomes from the TNF-α and NF-κB signaling pathways revealed single, multi-strain and Lh-R0052-SLP specific attenuation of the majority of proteins measured (TNF-α, IL-8, CXCL1, CXCL2 and CXCL10), indicating potentially different mechanisms. These findings indicate a synergistic effect of the bacterial combinations relative to the single strain and Lh-R0052-SLP treatments in resolving toll-like receptor 3 (TLR3)-induced inflammation in IEC and maintaining cellular homeostasis, reinforcing the rationale for using multi-strain formulations as a probiotic. PMID:28099447

  4. M5-branes on S^2 x M_4: Nahm's Equations and 4d Topological Sigma-models

    CERN Document Server

    Assel, Benjamin; Wong, Jin-Mann

    2016-01-01

    We study the 6d N=(0,2) superconformal field theory, which describes multiple M5-branes, on the product space S^2 x M_4, and suggest a correspondence between a 2d N=(0,2) half-twisted gauge theory on S^2 and a topological sigma-model on the four-manifold M_4. To set up this correspondence, we determine in this paper the dimensional reduction of the 6d N=(0,2) theory on a two-sphere and derive that the four-dimensional theory is a sigma-model into the moduli space of solutions to Nahm's equations, or equivalently the moduli space of k-centered SU(2) monopoles, where k is the number of M5-branes. We proceed in three steps: we reduce the 6d abelian theory to a 5d Super-Yang-Mills theory on I x M_4, with I an interval, then non-abelianize the 5d theory and finally reduce this to 4d. In the special case, when M_4 is a Hyper-Kahler manifold, we show that the dimensional reduction gives rise to a topological sigma-model based on tri-holomorphic maps. Deriving the theory on a general M_4 requires knowledge of the met...

  5. Discrimination and phylogenomic classification of Bacillus anthracis-cereus-thuringiensis strains based on LC-MS/MS analysis of whole cell protein digests.

    Science.gov (United States)

    Dworzanski, Jacek P; Dickinson, Danielle N; Deshpande, Samir V; Snyder, A Peter; Eckenrode, Brian A

    2010-01-01

    Modern taxonomy, diagnostics, and forensics of bacteria benefit from technologies that provide data for genome-based classification and identification of strains; however, full genome sequencing is still costly, lengthy, and labor intensive. Therefore, other methods are needed to estimate genomic relatedness among strains in an economical and timely manner. Although DNA-DNA hybridization and techniques based on genome fingerprinting or sequencing selected genes like 16S rDNA, gyrB, or rpoB are frequently used as phylogenetic markers, analyses of complete genome sequences showed that global measures of genome relatedness, such as the average genome conservation of shared genes, can provide better strain resolution and give phylogenies congruent with relatedness revealed by traditional phylogenetic markers. Bacterial genomes are characterized by a high gene density; therefore, we investigated the integration of mass spectrometry-based proteomic techniques with statistical methods for phylogenomic classification of bacterial strains. For this purpose, we used a set of well characterized Bacillus cereus group strains isolated from poisoned food to describe a method that relies on liquid chromatography-electrospray ionization-tandem mass spectrometry of tryptic peptides derived from whole cell digests. Peptides were identified and matched to a prototype database (DB) of reference bacteria with fully sequenced genomes to obtain their phylogenetic profiles. These profiles were processed for predicting genomic similarities with DB bacteria estimated by fractions of shared peptides (FSPs). FSPs served as descriptors for each food isolate and were jointly analyzed using hierarchical cluster analysis methods for revealing relatedness among investigated strains. The results showed that phylogenomic classification of tested food isolates was in consonance with results from established genomic methods, thus validating our findings. In conclusion, the proposed approach could be

  6. Some anomalous behaviour of vertebrates and insects preceding M5+ earthquakes in the North Western Apennines (Italy)

    Science.gov (United States)

    Straser, Valentino

    2013-04-01

    Earthquakes with a magnitude greater than M5+ are an unusual event in the seismic area of the Frignano District and the areas surrounding Parma in the North Western Apennines (Italy). Only two seismic events have occurred in the last four years: on 23 December 2008 (M5.1) and on 27 January 2012 (M5.4). The earthquake of 23 December 2008 allowed the verification of unusual behaviour in man and animals in the run-up to the main shock, in addition to anomalies of an electromagnetic type. An initial study showed that there are elements of coincidence between the seismic events and the number of admissions to hospitals around the epicentre: in the month of December 2008, the days with the greatest number of admissions coincided with seismic shocks. A half hour before the main event of 23 December, recorded at 16:24:21 local time (see: INGV), a slowworm (Anguis fragilis) left its hibernation site and died shortly afterwards from the cold on a road, as did a viper (Vipera aspis) found near some dwellings in an area around twenty kilometres from the epicentre. The investigation proceeded in 2009, but this time based on the number of daily admissions to the hospital A&E department, between June and December 2009. During the six months of the investigation, the maximum number of emergencies was 9 per day, while the earthquakes were in line with the usual number and magnitude for the Frignano seismic district. The earthquakes from June to December 2009 numbered 10, with a magnitude from M2.5 to M3.6. In 8 cases, in the 48 hours preceding the occurrence of the seism, there was a greater number of hospital emergencies. The subsequent occasion to check on a possible relationship between anomalous behaviour in animals and a seism occurred on 27 January 2012 (see: INGV), when an earthquake with a magnitude of M5.4 shook the North Western Apennines, thankfully without resulting in victims. Like 2008, in an area around fifteen kilometres from the epicentre, a grass snake (Zamenis

  7. Strain variability in fatty acid composition of Chattonella marina (Raphidophyceae) and its relation to differing ichthyotoxicity toward rainbow trout gill cells.

    Science.gov (United States)

    Dorantes-Aranda, Juan José; Nichols, Peter D; David Waite, Trevor; Hallegraeff, Gustaaf M

    2013-04-01

    Lipid profiles of three strains (Mexico, Australia, Japan) of Chattonella marina (Subrahmanyan) Hara et Chihara were studied under defined growth (phosphate, light, and growth phase) and harvest (intact and ruptured cells) conditions. Triacylglycerol levels were always 51% of total lipids). The major fatty acids in C. marina were palmitic (16:0), eicosapentaenoic (EPA, 20:5ω3), octadecatetraenoic (18:4ω3), myristic (14:0), and palmitoleic (16:1ω7c) acids. Higher levels of EPA were found in ruptured cells (21.4-29.4%) compared to intact cells (8.5-25.3%). In general, Japanese N-118 C. marina was the highest producer of EPA (14.3-29.4%), and Mexican CMCV-1 the lowest producer (7.9-27.1%). Algal cultures, free fatty acids from C. marina, and the two aldehydes 2E,4E-decadienal and 2E,4E-heptadienal (suspected fatty acid-derived products) were tested against the rainbow trout fish gill cell line RTgill-W1. The configuration of fatty acids plays an important role in ichthyotoxicity. Free fatty acid fractions, obtained by base saponification of total lipids from C. marina showed a potent toxicity toward gill cells (median lethal concentration, LC50 (at 1 h) of 0.44 μg · mL(-1) in light conditions, with a complete loss of viability at >3.2 μg · mL(-1) ). Live cultures of Mexican C. marina were less toxic than Japanese and Australian strains. This difference could be related to differing EPA content, superoxide anion production, and cell fragility. The aldehydes 2E,4E-decadienal and 2E,4E-heptadienal also showed high impact on gill cell viability, with LC50 (at 1 h) of 0.34 and 0.36 μg · mL(-1) , respectively. Superoxide anion production was highest in Australian strain CMPL01, followed by Japanese N-118 and Mexican CMCV-1 strains. Ruptured cells showed higher production of superoxide anion compared to intact cells (e.g., 19 vs. 9.5 pmol · cell(-1)  · hr(-1) for CMPL01, respectively). Our results indicate that C. marina is more ichthyotoxic

  8. Cytokine responses in primary chicken embryo intestinal cells infected with Campylobacter jejuni strains of human and chicken origin and the expression of bacterial virulence-associated genes

    Directory of Open Access Journals (Sweden)

    Bang Dang D

    2008-06-01

    Full Text Available Abstract Background Campylobacter jejuni is a major cause of inflammatory diarrhoea in humans and is considered a commensal of the gastroenteric tract of the avian host. However, little is known about the interaction between C. jejuni and the avian host including the cytokine responses and the expression of the bacterial genes. We have investigated the invasiveness of primary chicken embryo intestinal cells (CEICs by C. jejuni strains of human and chicken origins and the production of pro-inflammatory cytokines as well as the expression of the bacterial virulence-associated genes during co-cultivation. Results C. jejuni strains are capable of invading the CEICs and stimulate these cells in a pro-inflammatory manner and during this interaction the expression of the bacterial virulence-associated genes ciaB, dnaJ and racR is increased. Furthermore, incubation of bacteria with conditioned cell- and bacteria-free media from another co-cultivation experiment also increased the expression of the virulence-associated genes in the C. jejuni chicken isolate, indicating that the expression of bacterial genes is regulated by component(s secreted upon co-cultivation of bacteria and CEICs. Conclusion We show that under in vitro culture condition C. jejuni strains of both human and chicken origins can invade avian host cells with a pro-inflammatory response and that the virulence-associated genes of C. jejuni may play a role in this process.

  9. Heterologous expression and enzymatic properties of pullulanase from Klebsiella oxytoca M5 al%产酸克雷伯氏菌M5al普鲁兰酶的异源表达及酶学性质研究

    Institute of Scientific and Technical Information of China (English)

    温亮亮; 郭佳; 关锋

    2015-01-01

    Pullulanase(EC3. 2. 1. 41),one of debranching enzymes,can specifically hydrolyze α-1,6-glucosidic bonds in starch and greatly promote starch utilization. Pullulanase is widely used in food,textile,biofuel and detergent industry. In present study,pullulanase gene pulA was amplified by PCR using genomic DNA of Klebsiella oxytoca M5al as template, then cloned into vector pET28a(+)and expressed in Escherichia coli BL21 (DE3)induced by 0. 5 mmol/L IPTG. The expression of recombinant pullulanase was confirmed by SDS-PAGE and western blot. The enzyme was purified by Ni-Agarose resin and its enzymatic properties were determined. The optimum pH and temperature of recombinant enzyme were 5. 5 and 60 ° C respectively. The activity of recombinant enzyme was significantly increased while adding Mn2+,but slightly enhanced by adding Fe3+,Mg2+ and Fe2+. The enzyme activity was greatly inhibited in the presence of Cu2+. The optimal catalytic conditions of pullulanase from Klebsiella oxytoca M5 al met the requirement of saccharification process,showing its application potential in starch industry.%普鲁兰酶(EC 3.2.1.41)是一类淀粉脱支酶,能够特异性水解淀粉中的α-1,6-糖苷键,从而提高淀粉的利用率,在以淀粉为原料的食品、纺织、生物燃料和洗涤剂等行业中具有重要的应用价值。本研究以产酸克雷伯氏菌Klebsiella oxytoca M5 al基因组DNA为模板,将PCR扩增得到的普鲁兰酶基因pulA克隆至表达载体pET28 a (+),构建好的重组质粒转化大肠杆菌Escherichia coli BL21(DE3),在培养基中添加0.5 mmol/L异丙基硫代半乳糖苷(IPTG)的条件下对该酶基因进行诱导表达,经镍柱纯化获得重组普鲁兰酶用于酶学性质研究。SDS-PAGE及Western Blot检测显示普鲁兰酶基因pulA在上述大肠杆菌宿主中成功获得了表达。该重组酶最适反应pH5 .5 ,最适温度60℃。金属离子对酶活性有一定影响。Mn2+

  10. TheQc value of coda for EryuanM 5.5 earthquake in Yunan Province in the year 2013%2013年云南洱源M 5.5地震尾波Qc值

    Institute of Scientific and Technical Information of China (English)

    高琼; 李孝宾; 陈佳; 王军

    2016-01-01

    2013年3月3日云南洱源发生M 5.5地震,选取地震发生前后洱源地震台记录的40个数字化地震波形,基于地方震尾波单次散射模型,测算震源区尾波 Qc ( f )值。结果表明,当中心频率取3.0—18 Hz时,洱源地区尾波Qc值为181.08—692.88,得到Qc值与频率f的关系为Qc(f)=87.4f0.739,表明该区属中等构造活动地区。%AnM 5.5 earthquake occurred in Eryuan of Yunnan Province on Mar.3, 2013. Using forty digital data before and after Eryuan earthquake recorded by Eryuan Seismic Station, based on the single scattering theory of coda waves for local earthquakes, theQc value of coda at the source region has been estimated. The results show that theQc value of coda waves at Eryuan region is in the range of 181.08—692.88 when central frequency is in the range of 3.0—18.0 Hz. It was obtained that the relationship between codaQc and frequencyf isQc ( f ) = 87.4f0.739 in this area, so the seismic area belongs to moderate tectonic activity area.

  11. The 3-D aftershock distribution of three recent M5~5.5 earthquakes in the Anza region,California

    Science.gov (United States)

    Zhang, Q.; Wdowinski, S.; Lin, G.

    2011-12-01

    The San Jacinto fault zone (SJFZ) exhibits the highest level of seismicity compared to other regions in southern California. On average, it produces four earthquakes per day, most of them at depth of 10-17 km. Over the past decade, an increasing seismic activity occurred in the Anza region, which included three M5~5.5 events and their aftershock sequences. These events occurred in 2001, 2005, and 2010. In this research we map the 3-D distribution of these three events to evaluate their rupture geometry and better understand the unusual deep seismic pattern along the SJFZ, which was termed "deep creep" (Wdowinski, 2009). We relocated 97,562 events from 1981 to 2011 in Anza region by applying the Source-Specific Station Term (SSST) method (Lin et al., 2006) and used an accurate 1-D velocity model derived from 3-D model of Lin et al (2007) and used In order to separate the aftershock sequence from background seismicity, we characterized each of the three aftershock sequences using Omori's law. Preliminary results show that all three sequences had a similar geometry of deep elongated aftershock distribution. Most aftershocks occurred at depth of 10-17 km and extended over a 70 km long segments of the SJFZ, centered at the mainshock hypocenters. A comparative study of other M5~5.5 mainshocks and their aftershock sequences in southern California reveals very different geometrical pattern, suggesting that the three Anza M5~5.5 events are unique and can be indicative of "deep creep" deformation processes. Reference 1.Lin, G.and Shearer,P.M.,2006, The COMPLOC earthquake location package,Seism. Res. Lett.77, pp.440-444. 2.Lin, G. and Shearer, P.M., Hauksson, E., and Thurber C.H.,2007, A three-dimensional crustal seismic velocity model for southern California from a composite event method,J. Geophys.Res.112, B12306, doi: 10.1029/ 2007JB004977. 3.Wdowinski, S. ,2009, Deep creep as a cause for the excess seismicity along the San Jacinto fault, Nat. Geosci.,doi:10.1038/NGEO684.

  12. Rate of red blood cell destruction varies in different strains of mice infected with Plasmodium berghei-ANKA after chronic exposure

    Directory of Open Access Journals (Sweden)

    Kikuchi Mihoko

    2009-05-01

    Full Text Available Abstract Background Severe malaria anaemia in the semi-immune individuals in the holo-endemic area has been observed to occur at low parasite density with individual variation in the responses. Thus the following has been thought to be involved: auto-immune-mediated mechanisms of uninfected red blood cell destruction, and host genetic factors to explain the differences in individual responses under the same malaria transmission. In this study, the extent of red blood cell (RBC destruction in different strains of semi-immune mice model at relatively low parasitaemia was studied. Methodology To generate semi-immunity, four strains of mice were taken through several cycles of infection and treatment. By means of immunofluorescent assay and ELISA, sera were screened for anti-erythrocyte auto-antibodies, and their relationship with haematological parameters and parasitaemia in the strains of semi-immune mice was investigated. Results Upon challenge with Plasmodium berghei ANKA after generating semi-immune status, different mean percentage haemoglobin (Hb drop was observed in the mice strains (Balb/c = 47.1%; NZW = 30.05%; C57BL/6 = 28.44%; CBA = 25.1%, which occurred on different days for each strain (for Balb/c, mean period = 13.6 days; for C57BL/6, NZW, and CBA mean period = 10.6, 10.8, 10.9 days respectively. Binding of antibody to white ghost RBCs was observed in sera of the four strains of semi-immune mice by immunofluorescence. Mean percentage Hb drop per parasitaemia was highest in Balb/c (73.6, followed by C57BL/6 (8.6, CBA (6.9 and NZW (4.0, p = 0.0005. Consequently, auto-antibodies level to ghost RBC were correlated with degree of anaemia and were highest in Balb/c, when compared with the other strains, p Conclusion The results presented in this study seem to indicate that anti-RBC auto-antibodies may be involved in the destruction of uninfected RBC in semi-immune mice at relatively low parasite burden. Host genetic factors may also

  13. Effective actions and topological strings. Off-shell mirror symmetry and mock modularity of multiple M5-branes

    Energy Technology Data Exchange (ETDEWEB)

    Hecht, Michael

    2011-10-20

    This thesis addresses two different topics within the field of string theory. In the first part it is shown how Hodge-theoretic methods in conjunction with open string mirror symmetry can be used to compute non-perturbative effective superpotential couplings for type II/F-theory compactifications with D-branes and fluxes on compact Calabi-Yau manifolds. This is achieved by studying the at structure of operators which derives from the open/closed {beta}-model geometry. We analyze the variation of mixed Hodge structure of the relative cohomology induced by a family of divisors, which is wrapped by a D7-brane. This leads to a Picard-Fuchs system of differential operators, which can be used to compute the moduli dependence of the superpotential couplings as well as the mirror maps at various points in the open/closed deformation space. These techniques are used to obtain predictions for genuine A-model Ooguri-Vafa invariants of special Lagrangian submanifolds in compact Calabi-Yau geometries and real enumerative invariants of on-shell domain wall tensions. By an open/closed duality the system of differential equations can also be obtained from a gauged linear {sigma}-model, which describes a non-compact Calabi-Yau four-fold compactification without branes. This is used in the examples of multi-parameter models to study the various phases of the combined open/closed deformation space. It is furthermore shown how the brane geometry can be related to a F-theory compactification on a compact Calabi-Yau four-fold, where the Hodge-theoretic techniques can be used to compute the G-flux induced Gukov-Vafa-Witten potential. The dual F-theory picture also allows to conjecture the form of the Kaehler potential on the full open/closed deformation space. In the second part we analyze the background dependence of theories which derive from multiple wrapped M5-branes. Using the Kontsevich-Soibelman wall-crossing formula and the theory of mock modular forms we derive a holomorphic

  14. A kind of strain gauge load cell structure%一种电阻应蛮式称重传感器结构

    Institute of Scientific and Technical Information of China (English)

    张荣轩

    2012-01-01

    本文主要介绍了一种应用在称重式皮带给料机或称重式皮带配料秤产品上的平行梁型电阻应变式称重传感器创新结构。%This article mainly introduces a kind of parallel beam type strain gauge load cell innovation structure which is used in weighing belt feeding machine or weighing belt batch scale productions.

  15. Improved derivation efficiency and pluripotency of stem cells from the refractory inbred C57BL/6 mouse strain by small molecules.

    Science.gov (United States)

    Lin, Chih-Jen; Amano, Tomokazu; Tang, Yong; Tian, Xiuchun

    2014-01-01

    The ability of small molecules to maintain self-renewal and to inhibit differentiation of pluripotent stem cells has been well-demonstrated. Two widely used molecules are PD 98059 (PD), an inhibitor of extracellular-signal-regulated kinase 1 (ERK), and SC1 (Pluripotin), which inhibits the RasGAP and ERK pathways. However, no studies have been conducted to compare their effects on the pluripotency and derivation of embryonic stem (ES) cells from inbred mice C57BL/6, an important mouse strain frequently used to model behavior, cognitive functions, immune system, and metabolic disorders in humans and also the first mouse strain chosen to be sequenced for its entire genome. We found significantly increased derivation efficiency of ES cells from in vivo fertilized embryos (fES) of C57BL/6 with the use of PD (71.4% over the control of 35.3%). Because fES and ES from cloned embryos (ntES) are not distinguishable in transcription or translation profiles, we used ntES cells to compare the effect of small molecules on their in vitro characteristics, in vitro differentiation ability, and the ability to generate full-term ntES-4N pups by tetraploid complementation. NtES cells exhibited typical ES characteristics and up-regulated Sox2 expression in media with either small-molecule. Higher rates of full term ntES-4N pup were generated by the supplementation of PD or SC1. We obtained the highest efficiency of ntES-4N pup generation ever reported from this strain by supplementing ES medium with SC1. Lastly, we compared the pluripotency of fES, ntES and induced pluripotent stem (iPS) cells of C57BL/6 background using the tetraploid complementation assay. A significant increase in implantation sites and the number of full-term pups were obtained when fES, ntES, and iPS cells were cultured with SC1 compared to the control ES medium. In conclusion, supplementing ES cell culture medium with PD and SC1 increases the derivation efficiency and pluripotency, respectively, of stem cells

  16. M5-branes on S 2 × M 4: Nahm's equations and 4d topological sigma-models

    Science.gov (United States)

    Assel, Benjamin; Schäfer-Nameki, Sakura; Wong, Jin-Mann

    2016-09-01

    We study the 6d N = (0 , 2) superconformal field theory, which describes multiple M5-branes, on the product space S 2 × M 4, and suggest a correspondence between a 2d N = (0 , 2) half-twisted gauge theory on S 2 and a topological sigma-model on the four-manifold M 4. To set up this correspondence, we determine in this paper the dimensional reduction of the 6d N = (0 , 2) theory on a two-sphere and derive that the four-dimensional theory is a sigma-model into the moduli space of solutions to Nahm's equations, or equivalently the moduli space of k-centered SU(2) monopoles, where k is the number of M5-branes. We proceed in three steps: we reduce the 6d abelian theory to a 5d Super-Yang-Mills theory on I × M 4, with I an interval, then non-abelianize the 5d theory and finally reduce this to 4d. In the special case, when M 4 is a Hyper-Kähler manifold, we show that the dimensional reduction gives rise to a topological sigma-model based on tri-holomorphic maps. Deriving the theory on a general M 4 requires knowledge of the metric of the target space. For k = 2 the target space is the Atiyah-Hitchin manifold and we twist the theory to obtain a topological sigma-model, which has both scalar fields and self-dual two-forms.

  17. Characterization of the 2009 pandemic A/Beijing/501/2009 H1N1 influenza strain in human airway epithelial cells and ferrets.

    Directory of Open Access Journals (Sweden)

    Penghui Yang

    Full Text Available BACKGROUND: A novel 2009 swine-origin influenza A H1N1 virus (S-OIV H1N1 has been transmitted among humans worldwide. However, the pathogenesis of this virus in human airway epithelial cells and mammals is not well understood. METHODOLOGY/PRINCIPAL FINDING: In this study, we showed that a 2009 A (H1N1 influenza virus strain, A/Beijing/501/2009, isolated from a human patient, caused typical influenza-like symptoms including weight loss, fluctuations in body temperature, and pulmonary pathological changes in ferrets. We demonstrated that the human lung adenocarcinoma epithelial cell line A549 was susceptible to infection and that the infected cells underwent apoptosis at 24 h post-infection. In contrast to the seasonal H1N1 influenza virus, the 2009 A (H1N1 influenza virus strain A/Beijing/501/2009 induced more cell death involving caspase-3-dependent apoptosis in A549 cells. Additionally, ferrets infected with the A/Beijing/501/2009 H1N1 virus strain exhibited increased body temperature, greater weight loss, and higher viral titers in the lungs. Therefore, the A/Beijing/501/2009 H1N1 isolate successfully infected the lungs of ferrets and caused more pathological lesions than the seasonal influenza virus. Our findings demonstrate that the difference in virulence of the 2009 pandemic H1N1 influenza virus and the seasonal H1N1 influenza virus in vitro and in vivo may have been mediated by different mechanisms. CONCLUSION/SIGNIFICANCE: Our understanding of the pathogenesis of the 2009 A (H1N1 influenza virus infection in both humans and animals is broadened by our findings that apoptotic cell death is involved in the cytopathic effect observed in vitro and that the pathological alterations in the lungs of S-OIV H1N1-infected ferrets are much more severe.

  18. Analysis of the role of the LH92_11085 gene of a biofilm hyper-producing Acinetobacter baumannii strain on biofilm formation and attachment to eukaryotic cells.

    Science.gov (United States)

    Álvarez-Fraga, Laura; Pérez, Astrid; Rumbo-Feal, Soraya; Merino, María; Vallejo, Juan Andrés; Ohneck, Emily J; Edelmann, Richard E; Beceiro, Alejandro; Vázquez-Ucha, Juan C; Valle, Jaione; Actis, Luis A; Bou, Germán; Poza, Margarita

    2016-05-18

    Acinetobacter baumannii is a nosocomial pathogen that has a considerable ability to survive in the hospital environment partly due to its capacity to form biofilms. The first step in the process of establishing an infection is adherence of the bacteria to target cells. Chaperone-usher pili assembly systems are involved in pilus biogenesis pathways that play an important role in adhesion to host cells and tissues as well as medically relevant surfaces. After screening a collection of strains, a biofilm hyper-producing A. baumannii strain (MAR002) was selected to describe potential targets involved in pathogenicity. MAR002 showed a remarkable ability to form biofilm and attach to A549 human alveolar epithelial cells. Analysis of MAR002 using transmission electron microscopy (TEM) showed a significant presence of pili on the bacterial surface. Putative protein-coding genes involved in pili formation were identified based on the newly sequenced genome of MAR002 strain (JRHB01000001/2 or NZ_JRHB01000001/2). As assessed by qRT-PCR, the gene LH92_11085, belonging to the operon LH92_11070-11085, is overexpressed (ca. 25-fold more) in biofilm-associated cells compared to exponential planktonic cells. In the present work we investigate the role of this gene on the MAR002 biofilm phenotype. Scanning electron microscopy (SEM) and biofilm assays showed that inactivation of LH92_11085 gene significantly reduced bacterial attachment to A549 cells and biofilm formation on plastic, respectively. TEM analysis of the LH92_11085 mutant showed the absence of long pili formations normally present in the wild-type. These observations indicate the potential role this LH92_11085 gene could play in the pathobiology of A baumannii.

  19. Effects of gasket on coupled plastic flow and strain-induced phase transformations under high pressure and large torsion in a rotational diamond anvil cell

    Science.gov (United States)

    Feng, Biao; Levitas, Valery I.

    2016-01-01

    Combined plastic flow and strain-induced phase transformations (PTs) under high pressure in a sample within a gasket subjected to three dimensional compression and torsion in a rotational diamond anvil cell (RDAC) are studied using a finite element approach. The results are obtained for the weaker, equal-strength, and stronger high-pressure phases in comparison with low-pressure phases. It is found that, due to the strong gasket, the pressure in the sample is relatively homogenous and the geometry of the transformed zones is mostly determined by heterogeneity in plastic flow. For the equal-strength phases, the PT rate is higher than for the weaker and stronger high-pressure phases. For the weaker high-pressure phase, transformation softening induces material instability and leads to strain and PT localization. For the stronger high-pressure phase, the PT is suppressed by strain hardening during PT. The effect of the kinetic parameter k that scales the PT rate in the strain-controlled kinetic equation is also examined. In comparison with a traditional diamond anvil cell without torsion, the PT progress is much faster in RDAC under the same maximum pressure in the sample. Finally, the gasket size and strength effects are discussed. For a shorter and weaker gasket, faster plastic flow in radial and thickness directions leads to faster PT kinetics in comparison with a longer and stronger gasket. The rates of PT and plastic flows are not very sensitive to the modest change in a gasket thickness. Multiple experimental results are reproduced and interpreted. Obtained results allow one to design the desired pressure-plastic strain loading program in the experiments for searching new phases, reducing PT pressure by plastic shear, extracting kinetic properties from experiments with heterogeneous fields, and controlling homogeneity of all fields and kinetics of PTs.

  20. Influence of Additive Manufactured Scaffold Architecture on the Distribution of Surface Strains and Fluid Flow Shear Stresses and Expected Osteochondral Cell Differentiation.

    Science.gov (United States)

    Hendrikson, Wim J; Deegan, Anthony J; Yang, Ying; van Blitterswijk, Clemens A; Verdonschot, Nico; Moroni, Lorenzo; Rouwkema, Jeroen

    2017-01-01

    Scaffolds for regenerative medicine applications should instruct cells with the appropriate signals, including biophysical stimuli such as stress and strain, to form the desired tissue. Apart from that, scaffolds, especially for load-bearing applications, should be capable of providing mechanical stability. Since both scaffold strength and stress-strain distributions throughout the scaffold depend on the scaffold's internal architecture, it is important to understand how changes in architecture influence these parameters. In this study, four scaffold designs with different architectures were produced using additive manufacturing. The designs varied in fiber orientation, while fiber diameter, spacing, and layer height remained constant. Based on micro-CT (μCT) scans, finite element models (FEMs) were derived for finite element analysis (FEA) and computational fluid dynamics (CFD). FEA of scaffold compression was validated using μCT scan data of compressed scaffolds. Results of the FEA and CFD showed a significant impact of scaffold architecture on fluid shear stress and mechanical strain distribution. The average fluid shear stress ranged from 3.6 mPa for a 0/90 architecture to 6.8 mPa for a 0/90 offset architecture, and the surface shear strain from 0.0096 for a 0/90 offset architecture to 0.0214 for a 0/90 architecture. This subsequently resulted in variations of the predicted cell differentiation stimulus values on the scaffold surface. Fluid shear stress was mainly influenced by pore shape and size, while mechanical strain distribution depended mainly on the presence or absence of supportive columns in the scaffold architecture. Together, these results corroborate that scaffold architecture can be exploited to design scaffolds with regions that guide specific tissue development under compression and perfusion. In conjunction with optimization of stimulation regimes during bioreactor cultures, scaffold architecture optimization can be used to improve

  1. Recognition of tumor antigens in 4T1 cells by natural IgM from three strains of mice with different susceptibilities to spontaneous breast cancer

    Science.gov (United States)

    Díaz-Zaragoza, Mariana; Hernández-Ávila, Ricardo; Ostoa-Saloma, Pedro

    2017-01-01

    The issue of antibody responses to tumors is potentially important to cancer immunologists. Early detection of cancer represents one of the most promising approaches to reduce the growing cancer burden. Natural immunoglobulin (Ig)M antibodies have been associated with the recognition and elimination of cancerous and precancerous cells. Using natural IgM antibodies, the present study identified a set of antigens in healthy mice from three different strains and examined whether the global patterns of antibodies are able to discriminate between a condition of more or less susceptibility to breast cancer. The current study performed two-dimensional (2D) immunoblotting to detect antigens from 4T1 cells using natural IgM from serum of healthy female mice from three different strains. The t-test was used to analyze the total number of spots. There were no significant differences in the numbers of antigens recognized in each strain. However, differences in patterns were observed on 2D immunoblots among the three strains. The reactivity patterns of natural IgM antibodies to particular antigens exhibited non-random clustering, which discriminated between strains with different susceptibilities to spontaneous breast cancer. The results demonstrated that the patterns of reactivity to defined subsets of antigens are able to provide information regarding differential diagnosis associated with breast cancer sensitivity. Therefore, it may be concluded that it is possible to segregate the IgM humoral immune response toward cancer antigens according to the genetic background of individuals. In addition, it is possible to identify the recognized antigens that allow grouping or discriminate between the different IgM antibodies expressed. The possible association between a particular antigen and cancer susceptibility requires further study, but the methodology exposed in the present study may identify potential candidates for this possible association.

  2. Influence of Additive Manufactured Scaffold Architecture on the Distribution of Surface Strains and Fluid Flow Shear Stresses and Expected Osteochondral Cell Differentiation

    Science.gov (United States)

    Hendrikson, Wim J.; Deegan, Anthony J.; Yang, Ying; van Blitterswijk, Clemens A.; Verdonschot, Nico; Moroni, Lorenzo; Rouwkema, Jeroen

    2017-01-01

    Scaffolds for regenerative medicine applications should instruct cells with the appropriate signals, including biophysical stimuli such as stress and strain, to form the desired tissue. Apart from that, scaffolds, especially for load-bearing applications, should be capable of providing mechanical stability. Since both scaffold strength and stress–strain distributions throughout the scaffold depend on the scaffold’s internal architecture, it is important to understand how changes in architecture influence these parameters. In this study, four scaffold designs with different architectures were produced using additive manufacturing. The designs varied in fiber orientation, while fiber diameter, spacing, and layer height remained constant. Based on micro-CT (μCT) scans, finite element models (FEMs) were derived for finite element analysis (FEA) and computational fluid dynamics (CFD). FEA of scaffold compression was validated using μCT scan data of compressed scaffolds. Results of the FEA and CFD showed a significant impact of scaffold architecture on fluid shear stress and mechanical strain distribution. The average fluid shear stress ranged from 3.6 mPa for a 0/90 architecture to 6.8 mPa for a 0/90 offset architecture, and the surface shear strain from 0.0096 for a 0/90 offset architecture to 0.0214 for a 0/90 architecture. This subsequently resulted in variations of the predicted cell differentiation stimulus values on the scaffold surface. Fluid shear stress was mainly influenced by pore shape and size, while mechanical strain distribution depended mainly on the presence or absence of supportive columns in the scaffold architecture. Together, these results corroborate that scaffold architecture can be exploited to design scaffolds with regions that guide specific tissue development under compression and perfusion. In conjunction with optimization of stimulation regimes during bioreactor cultures, scaffold architecture optimization can be used to improve

  3. The effect of bacteriocin-producing Lactobacillus plantarum strains on the intracellular pH of sessile and planktonic Listeria monocytogenes single cells.

    Science.gov (United States)

    Nielsen, Dennis S; Cho, Gyu-Sung; Hanak, Alexander; Huch, Melanie; Franz, Charles M A P; Arneborg, Nils

    2010-07-31

    A wide range of lactic acid bacteria (LAB) produce bacteriocins mainly active against other closely related LAB, but some bacteriocins are also active against the food-borne pathogen Listeria monocytogenes. With the aim of increasing food safety it has thus been considered to utilise bacteriocins and/or bacteriocin-producing LAB as "natural" food preservatives in foods such as cheese, meat and ready-to-eat products. Some strains of Lactobacillus plantarum produce bacteriocins termed plantaricins. Using a single-cell based approach, the effect on the intracellular pH as a measure of the physiological state of sessile and planktonic L. monocytogenes (strains EGDe and N53-1) during co-culturing with plantaricin-producing L. plantarum (strains BFE 5092 and PCS 20) was investigated using fluorescence ratio imaging microscopy (FRIM). Mono-cultures of L. monocytogenes were used as control. Expression levels of plantaricin-encoding genes by sessile and planktonic L. plantarum were determined using qRT-PCR. L.plantarum BFE 5092 possesses the genes for plantaricin EF, JK and N, while L. plantarum PCS 20 contains the genes for plantaricin EF, although determination of the nucleotide sequence of the PCS 20 plantaricin E gene showed that this peptide is probably non-functional. When cultured as mono-culture, both L. monocytogenes strains maintained pH(i) at a constant level around 7.2-7.6 throughout the experiment, independently of the matrix. On a solid surface, L. plantarum BFE 5092 strongly affected pH(i) of L. monocytogenes N53-1 with only 20% of the cells being able to maintain pH(i) in the physiological optimal range with pH>7 and 52% of the cells with pH(i) approximately pH(ex,) showing that the cells had no proton gradient towards the environment. The effect on L. monocytogenes EGDe was less pronounced, but still notable. L.plantarum PCS 20 left both strains of L. monocytogenes virtually unaffected when co-cultured on a solid surface. In liquid, both L. plantarum

  4. Anaerobic and sequential aerobic production of high-titer ethanol and single cell protein from NaOH-pretreated corn stover by a genome shuffling-modified Saccharomyces cerevisiae strain.

    Science.gov (United States)

    Ren, Xueliang; Wang, Juncong; Yu, Hui; Peng, Chunlan; Hu, Jinlong; Ruan, Zhiyong; Zhao, Shumiao; Liang, Yunxiang; Peng, Nan

    2016-10-01

    In this study, a Saccharomyces cerevisiae recombinant strain 14 was constructed through genome shuffling method by transferring the whole genomic DNA of Candida intermedia strain 23 into a thermo-tolerant S. cerevisiae strain. The recombinant strain 14 combined the good natures of both parent strains that efficiently produced ethanol from glucose and single cell protein from xylose with 54.6% crude protein and all essential amino acids except cysteine at 35°C. Importantly, the recombinant strain 14 produced 64.07g/L ethanol from 25%(w/v) NaOH-pretreated and washed corn stover with the ethanol yield of 0.26g/g total stover by fed-batch simultaneous saccharification and fermentation and produced 66.50g/L dry cell mass subsequently from the residual hydrolysate and ethanol. Therefore, this study represents a feasible method to comprehensively utilize hexose and pentose in lignocellulosic materials.

  5. [Effect of mouse genotype on the hematopoietic stem cell count. II. The number of hematopietic stem cells in BALB/c and CC57BR strain mice differing by the level of endogenous colony formation].

    Science.gov (United States)

    Kozlov, V A

    1979-01-01

    The number of stem hematopoietic cells in the hematopoietic organs of mice of BALB/c and CC57BR strains and (CC57BRXBALB/c)F1 hybrids was studied by the method of exogenous colony-forming units. The assay of migration of stem cells from the bone marrow to the spleen was carried out. It was found that the spleen and the bone marrow of mice of the studied genotypes contain approximately the same relative number of hematopoietic stem cells. The number of stem cells which migrate from the bone marrow to the spleen is greater in the mice of BALB/c strain than in the CC57BR mice.

  6. Preservation of Bacillus firmus Strain 37 and Optimization of Cyclodextrin Biosynthesis by Cells Immobilized on Loofa Sponge

    Directory of Open Access Journals (Sweden)

    Cristiane Moriwaki

    2012-08-01

    Full Text Available The preservation of Bacillus firmus strain 37 cells by lyophilization was evaluated and response surface methodology (RSM was used to optimize the β-cyclodextrin (β-CD production by cells immobilized on loofa sponge. Interactions were studied with the variables temperature, pH and dextrin concentration using a central composite design (CCD. Immobilization time influence on β-CD production was also investigated. B. firmus strain 37 cells remained viable after one year of storage, showing that the lyophilization is a suitable method for preservation of the microorganism. From the three-dimensional diagrams and contour plots, the best conditions for β-CD production were determined: temperature 60 °C, pH 8, and 18% dextrin. Considering that the amount of dextrin was high, a new assay was carried out, in which dextrin concentrations of 10, 15, and 18% were tested and the temperature of 60 °C and pH 8 were maintained. The results achieved showed very small differences and therefore, for economic reasons, the use of 10% dextrin is suggested. Increasing the immobilization time of cells immobilized on synthetic sponge the β-CD production decreased and did not change for cells immobilized on loofa sponge. The results of this research are important for microorganism preservation and essential in the optimization of the biosynthesis of CD.

  7. Increased number of intestinal villous M cells in levamisole -pretreated weaned pigs experimentally infected with F4ac+ enterotoxigenic Escherichia coli strain

    Science.gov (United States)

    Valpotić, H.; Kovšca Janjatović, A.; Lacković, G.; Božić, F.; Dobranić, V.; Svoboda, D.; Valpotić, I.; Popović, M.

    2010-01-01

    Immunoprophylaxis of porcine postweaning colibacillosis (PWC) caused by enterotoxigenic Escherichia coli (ETEC) expressing F4 fimbriae is an unsolved problem. Just as ETEC strains can exploit intestinal microfold (M) cells as the entry portal for infection, their high transcytotic ability make them an attractive target for mucosally delivered vaccines, adjuvants and therapeutics. We have developed a model of parenteral/oral immunization of 4-weeks-old pigs with either levamisole or vaccine candidate F4ac+ non-ETEC strain to study their effects on de novo differentiation of antigen-sampling M cells. Identification, localization and morphometric quantification of cytokeratin 18 positive M cells in the ileal mucosa of 6-weeks-old pigs revealed that they were: 1) exclusively located within villous epithelial layer, 2) significantly numerous (P< 0.01) in levamisole pretreated/challenged pigs, and 3) only slightly, but not significantly numerous in vaccinated/challenged pigs compared with non-pretreated/challenged control pigs. The fact that levamisole may affect the M cells frequency by increasing their numbers, makes it an interesting adjuvant to study development of an effective M cell-targeted vaccine against porcine PWC. PMID:22073366

  8. Increased number of intestinal villous M cells in levamisole - pretreated weaned pigs experimentally infected with F4ac+ enterotoxigenic Escherichia coli strain

    Directory of Open Access Journals (Sweden)

    H. Valpotić

    2010-07-01

    Full Text Available Immunoprophylaxis of porcine postweaning colibacillosis (PWC caused by enterotoxigenic Escherichia coli (ETEC expressing F4 fimbriae is an unsolved problem. Just as ETEC strains can exploit intestinal microfold (M cells as the entry portal for infection, their high transcytotic ability make them an attractive target for mucosally delivered vaccines, adjuvants and therapeutics. We have developed a model of parenteral/oral immunization of 4-weeks-old pigs with either levamisole or vaccine candidate F4ac+ non-ETEC strain to study their effects on de novo differentiation of antigen-sampling M cells. Identification, localization and morphometric quantification of cytokeratin 18 positive M cells in the ileal mucosa of 6-weeks-old pigs revealed that they were: 1 exclusively located within villous epithelial layer, 2 significantly numerous (P< 0.01 in levamisole pretreated/challenged pigs, and 3 only slightly, but not significantly numerous in vaccinated/challenged pigs compared with non-pretreated/challenged control pigs. The fact that levamisole may affect the M cells frequency by increasing their numbers, makes it an interesting adjuvant to study development of an effective M cell-targeted vaccine against porcine PWC.

  9. Role of acinetobactin-mediated iron acquisition functions in the interaction of Acinetobacter baumannii strain ATCC 19606T with human lung epithelial cells, Galleria mellonella caterpillars, and mice.

    Science.gov (United States)

    Gaddy, Jennifer A; Arivett, Brock A; McConnell, Michael J; López-Rojas, Rafael; Pachón, Jerónimo; Actis, Luis A

    2012-03-01

    Acinetobacter baumannii, which causes serious infections in immunocompromised patients, expresses high-affinity iron acquisition functions needed for growth under iron-limiting laboratory conditions. In this study, we determined that the initial interaction of the ATCC 19606(T) type strain with A549 human alveolar epithelial cells is independent of the production of BasD and BauA, proteins needed for acinetobactin biosynthesis and transport, respectively. In contrast, these proteins are required for this strain to persist within epithelial cells and cause their apoptotic death. Infection assays using Galleria mellonella larvae showed that impairment of acinetobactin biosynthesis and transport functions significantly reduces the ability of ATCC 19606(T) cells to persist and kill this host, a defect that was corrected by adding inorganic iron to the inocula. The results obtained with these ex vivo and in vivo approaches were validated using a mouse sepsis model, which showed that expression of the acinetobactin-mediated iron acquisition system is critical for ATCC 19606(T) to establish an infection and kill this vertebrate host. These observations demonstrate that the virulence of the ATCC 19606(T) strain depends on the expression of a fully active acinetobactin-mediated system. Interestingly, the three models also showed that impairment of BasD production results in an intermediate virulence phenotype compared to those of the parental strain and the BauA mutant. This observation suggests that acinetobactin intermediates or precursors play a virulence role, although their contribution to iron acquisition is less relevant than that of mature acinetobactin.

  10. Interclonal variations in the molecular karyotype of Trypanosoma cruzi: chromosome rearrangements in a single cell-derived clone of the G strain.

    Science.gov (United States)

    Lima, Fabio Mitsuo; Souza, Renata Torres; Santori, Fábio Rinaldo; Santos, Michele Fernandes; Cortez, Danielle Rodrigues; Barros, Roberto Moraes; Cano, Maria Isabel; Valadares, Helder Magno Silva; Macedo, Andréa Mara; Mortara, Renato Arruda; da Silveira, José Franco

    2013-01-01

    Trypanosoma cruzi comprises a pool of populations which are genetically diverse in terms of DNA content, growth and infectivity. Inter- and intra-strain karyotype heterogeneities have been reported, suggesting that chromosomal rearrangements occurred during the evolution of this parasite. Clone D11 is a single-cell-derived clone of the T. cruzi G strain selected by the minimal dilution method and by infecting Vero cells with metacyclic trypomastigotes. Here we report that the karyotype of clone D11 differs from that of the G strain in both number and size of chromosomal bands. Large chromosomal rearrangement was observed in the chromosomes carrying the tubulin loci. However, most of the chromosome length polymorphisms were of small amplitude, and the absence of one band in clone D11 in relation to its reference position in the G strain could be correlated to the presence of a novel band migrating above or below this position. Despite the presence of chromosomal polymorphism, large syntenic groups were conserved between the isolates. The appearance of new chromosomal bands in clone D11 could be explained by chromosome fusion followed by a chromosome break or interchromosomal exchange of large DNA segments. Our results also suggest that telomeric regions are involved in this process. The variant represented by clone D11 could have been induced by the stress of the cloning procedure or could, as has been suggested for Leishmania infantum, have emerged from a multiclonal, mosaic parasite population submitted to frequent DNA amplification/deletion events, leading to a 'mosaic' structure with different individuals having differently sized versions of the same chromosomes. If this is the case, the variant represented by clone D11 would be better adapted to survive the stress induced by cloning, which includes intracellular development in the mammalian cell. Karyotype polymorphism could be part of the T. cruzi arsenal for responding to environmental pressure.

  11. Expression QTL mapping in regulatory and helper T cells from the BXD family of strains reveals novel cell-specific genes, gene-gene interactions and candidate genes for auto-immune disease

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    Alberts Rudi

    2011-12-01

    Full Text Available Abstract Background Regulatory T cells (Tregs play an essential role in the control of the immune response. Treg cells represent important targets for therapeutic interventions of the immune system. Therefore, it will be very important to understand in more detail which genes are specifically activated in Treg cells versus T helper (Th cells, and which gene regulatory circuits may be involved in specifying and maintaining Treg cell homeostasis. Results We isolated Treg and Th cells from a genetically diverse family of 31 BXD type recombinant inbred strains and the fully inbred parental strains of this family--C57BL/6J and DBA/2J. Subsequently genome-wide gene expression studies were performed from the isolated Treg and Th cells. A comparative analysis of the transcriptomes of these cell populations allowed us to identify many novel differentially expressed genes. Analysis of cis- and trans-expression Quantitative Trait Loci (eQTLs highlighted common and unique regulatory mechanisms that are active in the two cell types. Trans-eQTL regions were found for the Treg functional genes Nrp1, Stat3 and Ikzf4. Analyses of the respective QTL intervals suggested several candidate genes that may be involved in regulating these genes in Treg cells. Similarly, possible candidate genes were found which may regulate the expression of F2rl1, Ctla4, Klrb1f. In addition, we identified a focused group of candidate genes that may be important for the maintenance of self-tolerance and the prevention of allergy. Conclusions Variation of expression across the strains allowed us to find many novel gene-interaction networks in both T cell subsets. In addition, these two data sets enabled us to identify many differentially expressed genes and to nominate candidate genes that may have important functions for the maintenance of self-tolerance and the prevention of allergy.

  12. Insights into origins of Human T-cell Lymphotropic Virus Type 1 based on new strains from aboriginal people of Canada.

    Science.gov (United States)

    Andonov, Anton; Coulthart, Michael B; Pérez-Losada, Marcos; Crandall, Keith A; Posada, David; Padmore, Ruth; Giulivi, Antonio; Oger, Joel J; Peters, Andrew A; Dekaban, Gregory A

    2012-12-01

    The causes of the worldwide distribution of Human T-cell Lymphotropic Virus Type 1 (HTLV-1) remain incompletely understood, with competing hypotheses regarding the number and timing of events leading to intercontinental spread on historical and prehistoric timescales. Ongoing discovery of this virus in aboriginal populations of Asia and the Americas has been the main source of evidence for the latter. We conducted molecular phylogenetic and dating analyses for 13 newly reported HTLV-1 strains from Canada. We analyzed two full-length proviral genomes from aboriginal residents of Nunavut (an autonomous territory in Northern Canada including most of the Canadian Arctic), 11 long-terminal-repeat (LTR) sequences from aboriginal residents of British Columbia's Pacific coast, and 2 LTR sequences from non-aboriginal Canadians. Phylogenetic analysis demonstrated a well-supported affinity between the two Nunavut strains and two East Asian strains, suggesting the presence of an Asian-American sublineage within the widespread "transcontinental" subgroup A clade of HTLV-1 Cosmopolitan subtype a. This putative sublineage was estimated to be 5400-11,900 years in age, consistent with a long-term presence of HTLV-1 in aboriginal populations of the Canadian Arctic. Phylogenetic affinities of the other 11 Canadian HTLV-1 aboriginal strains were diverse, strengthening earlier evidence for multiple incursions of this virus into coastal aboriginal populations of British Columbia. Our results are consistent with the hypothesis of ancient presence of HTLV-1 in aboriginal populations of North America. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Immune Response Induced by an Immunodominant 60 kDa Glycoprotein of the Cell Wall of Sporothrix schenckii in Two Mice Strains with Experimental Sporotrichosis.

    Science.gov (United States)

    Alba-Fierro, Carlos A; Pérez-Torres, Armando; Toriello, Conchita; Pulido-Camarillo, Evelyn; López-Romero, Everardo; Romo-Lozano, Yolanda; Gutiérrez-Sánchez, Gerardo; Ruiz-Baca, Estela

    2016-01-01

    Cell wall (CW) components of fungus Sporothrix schenckii are the major inductors antigens of immune responses. The immunodominant 60 kDa glycoprotein (gp60) has been shown to be associated with the virulence of this fungus but its role in experimental sporotrichosis is unknown. In this work, the immunological effects of CW-purified gp60 were investigated in a model of experimental subcutaneous sporotrichosis in normal and gp60-preimmunized C57BL/6 and BALB/c mice strains which were then infected with S. schenckii conidia. Results showed that both mice strains use different cytokine profiles in order to fight S. schenckii infection; C57BL/6 mice seem to use a Th17 response while BALB/c mice tend to depend on a Th1 profile. Preimmunization with gp60 showed a downregulatory effect on the immune response since cytokines levels were diminished in both strains. There were no significant differences in the magnitude of dorsoplantar inflammation between gp60-preimmunized and nonimmunized mice of both strains. However, skin lesions due to the infection in gp60-preimmunized mice were more severe in BALB/c than in C57BL/6 mice, suggesting that the antigen exerts a higher downregulatory effect on the Th1 response.

  14. Immune Response Induced by an Immunodominant 60 kDa Glycoprotein of the Cell Wall of Sporothrix schenckii in Two Mice Strains with Experimental Sporotrichosis

    Directory of Open Access Journals (Sweden)

    Carlos A. Alba-Fierro

    2016-01-01

    Full Text Available Cell wall (CW components of fungus Sporothrix schenckii are the major inductors antigens of immune responses. The immunodominant 60 kDa glycoprotein (gp60 has been shown to be associated with the virulence of this fungus but its role in experimental sporotrichosis is unknown. In this work, the immunological effects of CW-purified gp60 were investigated in a model of experimental subcutaneous sporotrichosis in normal and gp60-preimmunized C57BL/6 and BALB/c mice strains which were then infected with S. schenckii conidia. Results showed that both mice strains use different cytokine profiles in order to fight S. schenckii infection; C57BL/6 mice seem to use a Th17 response while BALB/c mice tend to depend on a Th1 profile. Preimmunization with gp60 showed a downregulatory effect on the immune response since cytokines levels were diminished in both strains. There were no significant differences in the magnitude of dorsoplantar inflammation between gp60-preimmunized and nonimmunized mice of both strains. However, skin lesions due to the infection in gp60-preimmunized mice were more severe in BALB/c than in C57BL/6 mice, suggesting that the antigen exerts a higher downregulatory effect on the Th1 response.

  15. Host-dependent control of early regulatory and effector T-cell differentiation underlies the genetic susceptibility of RAG2-deficient mouse strains to transfer colitis.

    Science.gov (United States)

    Valatas, V; He, J; Rivollier, A; Kolios, G; Kitamura, K; Kelsall, B L

    2013-05-01

    De novo differentiation of CD4(+)Foxp3(+) regulatory T cells (induced (i) Tregs) occurs preferentially in the gut-associated lymphoid tissues (GALT). We addressed the contribution of background genetic factors in affecting the balance of iTreg, T helper type 1 (Th1), and Th17 cell differentiation in GALT in vivo following the transfer of naive CD4(+)CD45RB(high) T cells to strains of RAG2-deficient mice with differential susceptibility to inflammatory colitis. iTregs represented up to 5% of CD4(+) T cells in mesenteric lymph nodes of less-susceptible C57BL/6 RAG2(-/-) mice compared with <1% in highly susceptible C57BL/10 RAG2(-/-) mice 2 weeks following T-cell transfer before the onset of colitis. Early Treg induction was correlated inversely with effector cell expansion and the severity of colitis development, was controlled primarily by host and not T-cell-dependent factors, and was strongly associated with interleukin-12 (IL-12)/23 production by host CD11c(+)CD103(+) dendritic cells. These data highlight the importance of genetic factors regulating IL-12/23 production in controlling the balance between iTreg differentiation and effector-pathogenic CD4(+) T-cell expansion in lymphopenic mice and indicate a direct role for iTregs in the regulation of colonic inflammation in vivo.

  16. DUOX2 promotes the elimination of the Klebsiella pneumoniae strain K5 from T24 cells through the reactive oxygen species pathway.

    Science.gov (United States)

    Lu, Huixia; Wu, Qi; Yang, Huijun

    2015-08-01

    Dual oxidase 2 (DUOX2) plays a major role in host defense in intestinal and airway epithelial cells through the reactive oxygen species (ROS) pathway. Klebsiella pneumoniae is a uropathogen that causes urinary tract infections. It is not known whether DUOX2 plays a role in host defense in bladder cancer epithelial cells. It is also not known whether Klebsiella pneumoniae invades T24 human bladder carcinoma cells and whether DUOX2 plays a role in eliminating the Klebsiella pneumoniae strain K5 through the ROS pathway in T24 cells. Thus, in the present study, we aimed to investigate the infectious capability of the Klebsiella pneumoniae K5 strain and the immunity-promoting capability of DUOX2 in T24 cells. We quantified the number of viable intracellular bacteria using the plate count method. DUOX2 expression was evaluated by western blot analysis and reverse transcription-quantitative PCR (RT-qPCR) following treatment with or without multiple cytokines, phorbol 12-myristate 13-acetate (PMA), muramyl dipeptide (MDP), N-acetylmuramyl-D-alanyl-D-isoglutamine (MDP-DD), H2O2 inhibitor, catalase (CAT), the nicotinamide adenine dinucleotide phosphate-oxidase (NADPH) oxidase inhibitor, diphenyleneiodonium (DPI), or siRNA targeting DUOX2 (siDUOX2). The levels of ROS in the T24 cells infected with the K5 strain were examined following treatment with DPI, CAT or siDUOX2. Our results revealed that DUOX2 expression increased and the number of viable intracellular bacteria decreased in the T24 cells following infection with the K4 bacteria. Treatment with the cytokines and MDP and PMA also induced DUOX2 expression and decreased the number of viable intracellular bacteria. The levels of ROS also increased following treatment with the cytokines and MDP and PMA. However, when the cells were treated with the inhibitors (DPI or CAT), these effects were all reversed. Our data demonstrated that DUOX2 played an important role in innate immunity against bacterial cytoinvasion through the

  17. Application of Subspace Detection to the 6 November 2011 M5.6 Prague, Oklahoma Aftershock Sequence

    Science.gov (United States)

    McMahon, N. D.; Benz, H.; Johnson, C. E.; Aster, R. C.; McNamara, D. E.

    2015-12-01

    Subspace detection is a powerful tool for the identification of small seismic events. Subspace detectors improve upon single-event matched filtering techniques by using multiple orthogonal waveform templates whose linear combinations characterize a range of observed signals from previously identified earthquakes. Subspace detectors running on multiple stations can significantly increasing the number of locatable events, lowering the catalog's magnitude of completeness and thus providing extraord