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Genetic diversity of heat-labile toxin expressed by enterotoxigenic Escherichia coli strains isolated from humans

LASARO, M. A.; RODRIGUES, J. F.; MATHIAS-SANTOS, C.; GUTH, B. E. C.; BALAN, A.; SBROGIO-ALMEIDA, M. E.; FERREIRA, L. C. S.
Fonte: AMER SOC MICROBIOLOGY Publicador: AMER SOC MICROBIOLOGY
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
25.81%
The natural diversity of the eft operons, encoding the heat-labile toxin LT-I (LT), carried by enterotoxigenic Escherichia coli (ETEC) strains isolated from humans was investigated. For many years, LT was supposed to be represented by a rather conserved toxin, and one derivative, produced by the reference H10407 strain, was intensively studied either as a virulence factor or as a vaccine adjuvant. Amplicons encompassing the two LT-encoding genes (eltA and eltB) of 51 human-derived ETEC strains, either LT+ (25 strains) only or LT+/ST+ (26 strains), isolated from asymptomatic (24 strains) or diarrheic (27 strains) subjects, were subjected to restriction fragment length polymorphism (RFLP) analysis and DNA sequencing. Seven polymorphic RFLP types of the H10407 strain were detected with six (BsaI, DdeI, HhaI, HincII, HphI, and MspI) restriction enzymes. Additionally, the single-nucleotide polymorphic analysis revealed 50 base changes in the eft operon, including 21 polymorphic sites at eltA and 9 at eltB. Based on the deduced amino acid sequences, 16 LT types were identified, including LT1, expressed by the H10407 strain and 23 other strains belonging to seven different serotypes, and LT2, expressed by 11 strains of six different serotypes. In vitro experiments carried out with purified toxins indicated that no significant differences in GM1-binding affinity could be detected among LT1...

Characterization of the Operon Encoding the Alternative ςB Factor from Bacillus anthracis and Its Role in Virulence

Fouet, Agnès; Namy, Olivier; Lambert, Guillaume
Fonte: American Society for Microbiology Publicador: American Society for Microbiology
Tipo: Artigo de Revista Científica
Publicado em /09/2000 EN
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The operon encoding the general stress transcription factor ςB and two proteins of its regulatory network, RsbV and RsbW, was cloned from the gram-positive bacterium Bacillus anthracis by PCR amplification of chromosomal DNA with degenerate primers, by inverse PCR, and by direct cloning. The gene cluster was very similar to the Bacillus subtilis sigB operon both in the primary sequences of the gene products and in the order of its three genes. However, the deduced products of sequences upstream and downstream from this operon showed no similarity to other proteins encoded by the B. subtilis sigB operon. Therefore, the B. anthracis sigB operon contains three genes rather than eight as in B. subtilis. The B. anthracis operon is preceded by a ςB-like promoter sequence, the expression of which depends on an intact ςB transcription factor in B. subtilis. It is followed by another open reading frame that is also preceded by a promoter sequence similarly dependent on B. subtilis ςB. We found that in B. anthracis, both these promoters were induced during the stationary phase and induction required an intact sigB gene. The sigB operon was induced by heat shock. Mutants from which sigB was deleted were constructed in a toxinogenic and a plasmidless strain. These mutants differed from the parental strains in terms of morphology. The toxinogenic sigB mutant strain was also less virulent than the parental strain in the mouse model. B. anthracis ςB may therefore be a minor virulence factor.

Amplification of the housekeeping sigma factor in Pseudomonas fluorescens CHA0 enhances antibiotic production and improves biocontrol abilities.

Schnider, U; Keel, C; Blumer, C; Troxler, J; Défago, G; Haas, D
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /09/1995 EN
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25.81%
Pseudomonas fluorescens CHA0 produces a variety of secondary metabolites, in particular the antibiotics pyoluteorin and 2,4-diacetylphloroglucinol, and protects various plants from diseases caused by soilborne pathogenic fungi. The rpoD gene encoding the housekeeping sigma factor sigma 70 of P. fluorescens was sequenced. The deduced RpoD protein showed 83% identity with RpoD of Pseudomonas aeruginosa and 67% identity with RpoD of Escherichia coli. Attempts to inactivate the single chromosomal rpoD gene of strain CHA0 were unsuccessful, indicating an essential role of this gene. When rpoD was carried by an IncP vector in strain CHA0, the production of both antibiotics was increased severalfold and, in parallel, protection of cucumber against disease caused by Pythium ultimum was improved, in comparison with strain CHA0.

Evidence that the potyvirus P1 proteinase functions in trans as an accessory factor for genome amplification.

Verchot, J; Carrington, J C
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /06/1995 EN
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25.85%
The tobacco etch potyvirus (TEV) polyprotein is proteolytically processed by three viral proteinases (NIa, HC-Pro, and P1). While the NIa and HC-Pro proteinases each provide multiple functions essential for viral infectivity, the role of the P1 proteinase beyond its autoproteolytic activity is understood poorly. To determine if P1 is necessary for genome amplification and/or virus movement from cell to cell, a mutant lacking the entire P1 coding region (delta P1 mutant) was produced with a modified TEV strain (TEV-GUS) expressing beta-glucuronidase (GUS) as a reporter, and its replication and movement phenotypes were assayed in tobacco protoplasts and plants. The delta P1 mutant accumulated in protoplasts to approximately 2 to 3% the level of parental TEV-GUS, indicating that the P1 protein may contribute to but is not strictly required for viral RNA amplification. The delta P1 mutant was capable of cell-to-cell and systemic (leaf-to-leaf) movement in plants but at reduced rates compared with parental virus. This is in contrast to the S256A mutant, which encodes a processing-defective P1 proteinase and which was nonviable in plants. Both delta P1 and S256A mutants were complemented by P1 proteinase expressed in a transgenic host. In transgenic protoplasts...

Patient-to-patient spread of a single strain of Corynebacterium striatum causing infections in a surgical intensive care unit.

Brandenburg, A H; van Belkum, A; van Pelt, C; Bruining, H A; Mouton, J W; Verbrugh, H A
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /09/1996 EN
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25.8%
Over a 12-month period, Corynebacterium striatum strains were isolated from clinical specimens from 14 patients admitted to a surgical intensive care unit. These isolates were identical by morphology and biotype and displayed the same antibiogram. Ten isolates were found to be the sole possible pathogen. These 10 isolates were from six patients, three of whom had signs of infection at the time of positive culture. Further typing was performed by random amplification of polymorphic DNA analysis, by which all strains were identical and were found to differ to various degrees from reference strains and from isolates found in clinical samples from other wards. In a case-control study the only independent risk factor for acquiring the strain was intubation for longer than 24 h (odds ratio, 20.09; 95% confidence interval, 2.29 to 176.09). The same strain was isolated from surfaces and from air sampled in the direct vicinity of infected patients but never from surfaces or air in other places of the ward. The strain was not isolated from the ventilators. The strain was cultured from the hands of personnel attending to infected patients, but no long-term carriers were found among members of the hospital personnel, suggesting transient carriage only. We conclude that C. striatum can cause serious nosocomial infections in surgical intensive care unit patients and may spread from patient to patient via the hands of attending personnel.

Human immunodeficiency virus-1 infection of macrophages in vitro neither induces tumor necrosis factor (TNF)/cachectin gene expression nor alters TNF/cachectin induction by lipopolysaccharide.

Munis, J R; Richman, D D; Kornbluth, R S
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /02/1990 EN
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25.77%
The synthesis of tumor necrosis factor (TNF)/cachectin was assessed in primary monocyte-derived macrophage (MDM) cultures after in vitro infection with a macrophage-tropic strain of HIV-1 (HTLV-IIIBa-L/85). Productive and cytopathic infections in MDM cultures were established using a high multiplicity of infection (m.o.i. = 3) under conditions that minimized endotoxin contamination. Culture supernatants were tested for TNF/cachectin activity by L929 cell cytotoxicity assay, and TNF/cachectin mRNA was assessed by a sensitive PCR amplification technique that could detect between 1 and 10 cells fully activated for TNF/cachectin expression. Unstimulated MDM cultures produced no detectable levels of TNF/cachectin activity or mRNA, consistent with previous demonstrations that production of this cytokine by macrophages is an inducible and not a constitutive event. HIV-1 infection failed to induce detectable TNF/cachectin activity or mRNA in these unstimulated cultures. In addition, the responsiveness of macrophages to lipopolysaccharide (LPS) induction of TNF/cachectin production was assessed in dose-response and kinetic experiments. No differences between infected and uninfected cultures were discernable. These results demonstrate that productive and cytopathic infection with a macrophage-tropic strain of HIV-1 does not alter the regulation of TNF/cachectin expression in macrophages.

Self-limiting nature of seasonal cholera epidemics: Role of host-mediated amplification of phage

Faruque, Shah M.; Islam, M. Johirul; Ahmad, Qazi Shafi; Faruque, A. S. G.; Sack, David A.; Nair, G. Balakrish; Mekalanos, John J.
Fonte: National Academy of Sciences Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
EN
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25.94%
Phage predation of Vibrio cholerae has recently been reported to be a factor that influences seasonal epidemics of cholera in Bangladesh. To understand more about this phenomenon, we studied the dynamics of the V. cholerae-phage interaction during a recent epidemic in Dhaka. Because the outbreak strain causing this epidemic was resistant to multiple antibiotics, including streptomycin, we used a selective medium containing streptomycin to monitor accurately the abundance of this strain in the environment. The changing prevalence in the environment of the epidemic V. cholerae O1 strain and a particular lytic cholera phage (JSF4) to which it was sensitive was measured every 48-72 h for 17 weeks. We also monitored the incidence of phage excretion in stools of 387 cholera patients during the epidemic. The peak of the epidemic was preceded by high V. cholerae prevalence in the environment and was followed by high JSF4 phage levels as the epidemic ended. The buildup to the phage peak in the environment coincided with increasing excretion of the same phage in the stools of cholera patients. These results suggest that patients toward the end of the epidemic ingested both JSF4 phage and the outbreak V. cholerae strain. Host-mediated phage amplification during the cholera epidemic likely contributed to increased environmental phage abundance...

Amplified Expression of Fructose 1,6-Bisphosphatase in Corynebacterium glutamicum Increases In Vivo Flux through the Pentose Phosphate Pathway and Lysine Production on Different Carbon Sources

Becker, Judith; Klopprogge, Corinna; Zelder, Oskar; Heinzle, Elmar; Wittmann, Christoph
Fonte: American Society for Microbiology Publicador: American Society for Microbiology
Tipo: Artigo de Revista Científica
Publicado em /12/2005 EN
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25.75%
The overexpression of fructose 1,6-bisphosphatase (FBPase) in Corynebacterium glutamicum leads to significant improvement of lysine production on different sugars. Amplified expression of FBPase via the promoter of the gene encoding elongation factor TU (EFTU) increased the lysine yield in the feedback-deregulated lysine-producing strain C. glutamicum lysCfbr by 40% on glucose and 30% on fructose or sucrose. Additionally formation of the by-products glycerol and dihydroxyacetone was significantly reduced in the PEFTUfbp mutant. As revealed by 13C metabolic flux analysis on glucose the overexpression of FBPase causes a redirection of carbon flux from glycolysis toward the pentose phosphate pathway (PPP) and thus leads to increased NADPH supply. Normalized to an uptake flux of glucose of 100%, the relative flux into the PPP was 56% for C. glutamicum lysCfbr PEFTUfbp and 46% for C. glutamicum lysCfbr. The flux for NADPH supply was 180% in the PEFTUfbp strain and only 146% in the parent strain. Amplification of FBPase increases the production of lysine via an increased supply of NADPH. Comparative studies with another mutant containing the sod promoter upstream of the fbp gene indicate that the expression level of FBPase relates to the extent of the metabolic effects. The overexpression of FBPase seems useful for starch- and molasses-based industrial lysine production with C. glutamicum. The redirection of flux toward the PPP should also be interesting for the production of other NADPH-demanding compounds as well as for products directly stemming from the PPP.

Deletion of Complement Factor H–Related Genes CFHR1 and CFHR3 Is Associated with Atypical Hemolytic Uremic Syndrome

Zipfel, Peter F; Edey, Matthew; Heinen, Stefan; Józsi, Mihály; Richter, Heiko; Misselwitz, Joachim; Hoppe, Bernd; Routledge, Danny; Strain, Lisa; Hughes, Anne E; Goodship, Judith A; Licht, Christoph; Goodship, Timothy H. J; Skerka, Christine
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
EN
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35.77%
Atypical hemolytic uremic syndrome (aHUS) is associated with defective complement regulation. Disease-associated mutations have been described in the genes encoding the complement regulators complement factor H, membrane cofactor protein, factor B, and factor I. In this study, we show in two independent cohorts of aHUS patients that deletion of two closely related genes, complement factor H–related 1 (CFHR1) and complement factor H–related 3 (CFHR3), increases the risk of aHUS. Amplification analysis and sequencing of genomic DNA of three affected individuals revealed a chromosomal deletion of ∼84 kb in the RCA gene cluster, resulting in loss of the genes coding for CFHR1 and CFHR3, but leaving the genomic structure of factor H intact. The CFHR1 and CFHR3 genes are flanked by long homologous repeats with long interspersed nuclear elements (retrotransposons) and we suggest that nonallelic homologous recombination between these repeats results in the loss of the two genes. Impaired protection of erythrocytes from complement activation is observed in the serum of aHUS patients deficient in CFHR1 and CFHR3, thus suggesting a regulatory role for CFHR1 and CFHR3 in complement activation. The identification of CFHR1/CFHR3 deficiency in aHUS patients may lead to the design of new diagnostic approaches...

The dynamic mechanical environment of the chondrocyte: A biphasic finite element model of cell-matrix interactions under cyclic compressive loading

Kim, Eunjung; Guilak, Farshid; Haider, Mansoor A.
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /12/2008 EN
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25.86%
Cyclic mechanical loading of articular cartilage results in a complex biomechanical environment at the scale of the chondrocytes that strongly affects cellular metabolic activity. Under dynamic loading conditions, the quantitative relationships between macroscopic loading characteristics and solid and fluid mechanical variables in the local cellular environment are not well understood. In this study, an axisymmetric multiscale model of linear biphasic cell-matrix interactions in articular cartilage was developed to investigate the cellular microenvironment in an explant subjected to cyclic confined compressive loading. The model was based on the displacement-velocity-pressure (u-v-p) mixed penalty weighted residual formulation of linear biphasic theory that was implemented in the COMSOL Multiphysics software package. The microscale cartilage environment was represented as a three-zone biphasic region consisting of a spherical chondrocyte with encapsulating pericellular matrix (PCM) that was embedded in a cylindrical extracellular matrix (ECM) subjected to cyclic confined compressive loading boundary conditions. Biphasic material properties for the chondrocyte and the PCM were chosen based on previous in vitro micropipette studies of cells or chondrons isolated from normal or osteoarthritic cartilage. Simulations performed at four loading frequencies in the range 0.01–1.0 Hz supported the hypothesized dual role of the PCM as both a protective layer for the cell and a mechanical transducer of strain. Time varying biphasic variables at the cellular scale were strongly dependent on relative magnitudes of the loading period...

Association of factor H autoantibodies with deletions of CFHR1, CFHR3, CFHR4, and with mutations in CFH, CFI, CD46, and C3 in patients with atypical hemolytic uremic syndrome

Moore, Iain; Strain, Lisa; Pappworth, Isabel; Kavanagh, David; Barlow, Paul N.; Herbert, Andrew P.; Schmidt, Christoph Q.; Staniforth, Scott J.; Holmes, Lucy V.; Ward, Roy; Morgan, Lynn; Goodship, Timothy H. J.; Marchbank, Kevin J.
Fonte: American Society of Hematology Publicador: American Society of Hematology
Tipo: Artigo de Revista Científica
Publicado em 14/01/2010 EN
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35.77%
Factor H autoantibodies have been reported in approximately 10% of patients with atypical hemolytic uremic syndrome (aHUS) and are associated with deficiency of factor H–related proteins 1 and 3. In this study we examined the prevalence of factor H autoantibodies in the Newcastle cohort of aHUS patients, determined whether the presence of such autoantibodies is always associated with deficiency of factor H–related proteins 1 and 3, and examined whether such patients have additional susceptibility factors and/or mutations in the genes encoding complement regulator/activators. We screened 142 patients with aHUS and found factor H autoantibodies in 13 individuals (age 1-11 years). The presence of the autoantibodies was confirmed by Western blotting. By using multiplex ligation-dependent probe amplification we measured complement factor H–related (CFHR)1 and CFHR3 copy number. In 10 of the 13 patients there were 0 copies of CFHR1, and in 3 patients there were 2. In 3 of the patients with 0 copies of CFHR1 there was 1 copy of CFHR3, and these individuals exhibited a novel deletion incorporating CFHR1 and CFHR4. In 5 patients mutations were identified: 1 in CFH, 1 in CFI, 1 in CD46, and 2 in C3. The latter observation emphasizes that multiple concurrent factors may be necessary in individual patients for disease manifestation.

Specific Role of the Cyanobacterial PipX Factor in the Heterocysts of Anabaena sp. Strain PCC 7120▿

Valladares, Ana; Rodríguez, Virginia; Camargo, Sergio; Martínez-Noël, Giselle M. A.; Herrero, Antonia; Luque, Ignacio
Fonte: American Society for Microbiology (ASM) Publicador: American Society for Microbiology (ASM)
Tipo: Artigo de Revista Científica
EN
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25.8%
The PipX factor is a regulatory protein that seems to occur only in cyanobacteria. In the filamentous, heterocyst-forming Anabaena sp. strain PCC 7120, open reading frame (ORF) asr0485, identified as the pipX gene, is expressed mainly under conditions of combined-nitrogen deprivation dependent on the global N regulator NtcA and the heterocyst-specific regulator HetR. Primer extension and 5′ rapid amplification of cDNA ends (RACE) analyses detected three transcription start points corresponding to a canonical NtcA-activated promoter (to which direct binding of NtcA was observed), an NtcA- and HetR-dependent promoter, and a consensus-type promoter, the last with putative −35 and −10 determinants. Activation of pipX took place in cells differentiating into heterocysts at intermediate to late stages of the process. Accordingly, disruption of pipX led to impaired diazotrophic growth, reduced nitrogenase activity, and impaired activation of the nitrogenase structural genes. The nitrogenase activity of the mutant was low under oxic conditions, likely resulting from inefficient protection against oxygen. In line with this, the activation of the coxB2A2C2 and coxB3A3C3 operons, encoding heterocyst-specific terminal respiratory oxidases responsible for internal oxygen removal...

Quaternary Structure of Pathological Prion Protein as a Determining Factor of Strain-Specific Prion Replication Dynamics

Laferrière, Florent; Tixador, Philippe; Moudjou, Mohammed; Chapuis, Jérôme; Sibille, Pierre; Herzog, Laetitia; Reine, Fabienne; Jaumain, Emilie; Laude, Hubert; Rezaei, Human; Béringue, Vincent
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
EN
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25.87%
Prions are proteinaceous infectious agents responsible for fatal neurodegenerative diseases in animals and humans. They are essentially composed of PrPSc, an aggregated, misfolded conformer of the ubiquitously expressed host-encoded prion protein (PrPC). Stable variations in PrPSc conformation are assumed to encode the phenotypically tangible prion strains diversity. However the direct contribution of PrPSc quaternary structure to the strain biological information remains mostly unknown. Applying a sedimentation velocity fractionation technique to a panel of ovine prion strains, classified as fast and slow according to their incubation time in ovine PrP transgenic mice, has previously led to the observation that the relationship between prion infectivity and PrPSc quaternary structure was not univocal. For the fast strains specifically, infectivity sedimented slowly and segregated from the bulk of proteinase-K resistant PrPSc. To carefully separate the respective contributions of size and density to this hydrodynamic behavior, we performed sedimentation at the equilibrium and varied the solubilization conditions. The density profile of prion infectivity and proteinase-K resistant PrPSc tended to overlap whatever the strain, fast or slow...

Genotypic Analysis of Meningococcal Factor H-Binding Protein from Non-Culture Clinical Specimens

Clark, Stephen A.; Lucidarme, Jay; Newbold, Lynne S.; Borrow, Ray
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em 24/02/2014 EN
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25.8%
Factor H-Binding Protein (fHbp) is an outer membrane protein antigen included in two novel meningococcal group B vaccines and, as such, is an important typing target. Approximately 50% of meningococcal disease cases in England and Wales are confirmed using real-time PCR on non-culture clinical specimens only. Protocols for typing fHbp from this subset of cases have not yet been established. Here we present a nested PCR-based assay designed to amplify and sequence fHbp from non-culture clinical specimens. From analytical sensitivity experiments carried out using diluted DNA extracts, an estimated analytical sensitivity limit of 6 fg/µL of DNA (<3 genome copies/µL) was calculated. The sensitivity of the assay was shown to be comparable to the ctrA-directed real-time PCR assay currently used to confirm invasive disease diagnoses from submitted clinical specimens. A panel of 96 diverse, patient-matched clinical specimen/isolate pairs from invasive disease cases was used to illustrate the breadth of strain coverage for the assay. All fHbp alleles sequenced from the isolates matched those derived from previous whole genome analyses. The first-round PCR primer binding sites are highly conserved, however an exceptional second-round PCR primer site mismatch in one validation isolate prevented amplification. In this case...

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

Milner, Jaques S.; Grol, Matthew W.; Beaucage, Kim L.; Dixon, S. Jeffrey; Holdsworth, David W.
Fonte: MDPI Publicador: MDPI
Tipo: Artigo de Revista Científica
Publicado em 22/03/2012 EN
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35.91%
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.

Strain Amplification Analysis of an Osteocyte under Static and Cyclic Loading: A Finite Element Study

Wang, Liping; Dong, Jianghui; Xian, Cory J.
Fonte: Hindawi Publishing Corporation Publicador: Hindawi Publishing Corporation
Tipo: Artigo de Revista Científica
EN
Relevância na Pesquisa
56.11%
Osteocytes, the major type of bone cells which reside in their lacunar and canalicular system within the bone matrix, function as biomechanosensors and biomechanotransducers of the bone. Although biomechanical behaviour of the osteocyte-lacunar-canalicular system has been investigated in previous studies mostly using computational 2-dimensional (2D) geometric models, only a few studies have used the 3-dimensional (3D) finite element (FE) model. In the current study, a 3D FE model was used to predict the responses of strain distributions of osteocyte-lacunar-canalicular system analyzed under static and cyclic loads. The strain amplification factor was calculated for all simulations. Effects on the strain of the osteocyte system were investigated under 500, 1500, 2000, and 3000 microstrain loading magnitudes and 1, 5, 10, 40, and 100 Hz loading frequencies. The maximum strain was found to change with loading magnitude and frequency. It was observed that maximum strain under 3000-microstrain loading was higher than those under 500, 1500, and 2000 microstrains. When the loading strain reached the maximum magnitude, the strain amplification factor of 100 Hz was higher than those of the other frequencies. Data from this 3D FE model study suggests that the strain amplification factor of the osteocyte-lacunar-canalicular system increases with loading frequency and loading strain increasing.

Effect of the microstructure parameters on the Mullins softening in carbon-black filled SBRs

MERCKEL, Yannick; DIANI, Julie; BRIEU, Matthias; GILORMINI, Pierre; CAILLARD, Julien
Fonte: Wiley Publicador: Wiley
EN
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66.09%
[The definitive version is available at www3.interscience.wiley.com]; A quantitative estimate of the Mullins softening is proposed and tested on various carbon-black filled styrene-butadiene rubbers. In order to model the behaviour of elastomeric materials, some constitutive equations reported in the literature are based on the account of a strain amplification factor, which evolves with the maximum strain history. The amplification factor is grounded on the representation of filled rubbers as heterogeneous materials made of hard rigid domains and soft deformable domains. In the present work, this factor is splitted into two parts with opposite effects that account for the Mullins softening and for the filler reinforcement, respectively. Evolutions of both parts are obtained through a direct analysis of cyclic uniaxial tensile tests performed on a series of materials. The Mullins softening part is shown to linearly depend on the filler volume fraction and on the maximum strain applied, when defined as the first invariant of the Hencky tensor. Its changes with the gum crosslink density parameter are insignificant. The reinforcement part of the amplification factor shows quadratic dependence on the filler volume fraction.; ANR MATETPRO AMUFISE

Charcaterization of the Mullins effect of carbon-black filled rubbers

MERCKEL, Yannick; DIANI, Julie; BRIEU, Matthias; GILORMINI, Pierre; CAILLARD, Julien
Fonte: American Chemical Society Publicador: American Chemical Society
EN
Relevância na Pesquisa
35.71%
Publisher version : http://rubberchemtechnol.org/doi/abs/10.5254/1.3592294?journalCode=rcat; Several carbon-black filled styrene-butadiene rubbers showed different sensibilities to the Mullins softening when submitted to cyclic uniaxial tension. In order to quantify this softening, a damage parameter was introduced. It is defined by using a classic damage approach and can be estimated by using either the strain amplification factor method or the tangent modulus at zero stress. The proposed parameter is used to study the effects of crosslink density and filler amount on the Mullins softening. The latter is shown to remain unaffected by a change of crosslink density and to increase with an increase of filler amount. The damage parameter exhibits mere linear dependences on the maximum Hencky strain applied and on the filler volume fraction. A simple linear expression is given finally to predict the Mullins softening of filled rubbers. The parameter also provides an objective analysis for the Mullins softening that supports comments on a better understanding of this effect.; ANR MATETPRO AMUFISE

The Plasmid of Escherichia coli Strain S88 (O45:K1:H7) That Causes Neonatal Meningitis Is Closely Related to Avian Pathogenic E. coli Plasmids and Is Associated with High-Level Bacteremia in a Neonatal Rat Meningitis Model▿

Peigne, Chantal; Bidet, Philippe; Mahjoub-Messai, Farah; Plainvert, Céline; Barbe, Valérie; Médigue, Claudine; Frapy, Eric; Nassif, Xavier; Denamur, Erick; Bingen, Edouard; Bonacorsi, Stéphane
Fonte: American Society for Microbiology (ASM) Publicador: American Society for Microbiology (ASM)
Tipo: Artigo de Revista Científica
EN
Relevância na Pesquisa
25.75%
A new Escherichia coli virulent clonal group, O45:K1, belonging to the highly virulent subgroup B21 was recently identified in France, where it accounts for one-third of E. coli neonatal meningitis cases. Here we describe the sequence, epidemiology and function of the large plasmid harbored by strain S88, which is representative of the O45:K1 clonal group. Plasmid pS88 is 133,853 bp long and contains 144 protein-coding genes. It harbors three different iron uptake systems (aerobactin, salmochelin, and the sitABCD genes) and other putative virulence genes (iss, etsABC, ompTP, and hlyF). The pS88 sequence is composed of several gene blocks homologous to avian pathogenic E. coli plasmids pAPEC-O2-ColV and pAPEC-O1-ColBM. PCR amplification of 11 open reading frames scattered throughout the plasmid was used to investigate the distribution of pS88 and showed that a pS88-like plasmid is present in other meningitis clonal groups such as O18:K1, O1:K1, and O83:K1. A pS88-like plasmid was also found in avian pathogenic strains and human urosepsis strains belonging to subgroup B21. A variant of S88 cured of its plasmid displayed a marked loss of virulence relative to the wild-type strain in a neonatal rat model, with bacteremia more than 2 log CFU/ml lower. The salmochelin siderophore...

Detection of Aeromonas salmonicida from fish by using polymerase chain reaction amplification of the virulence surface array protein gene.

Gustafson, C E; Thomas, C J; Trust, T J
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /12/1992 EN
Relevância na Pesquisa
25.74%
A DNA-based assay was developed to detect Aeromonas salmonicida from infected fish by analyzing tissues, feces, and the tank water in which the infected fish were held. This analysis was done both by direct detection from samples and after a bacterial outgrowth step. Polymerase chain reaction (PCR) amplification of a 421-bp sequence from the 3' region of the surface array protein gene (vapA) of A. salmonicida provided a specific and sensitive method for the detection and identification of this important fish pathogen. The sensitivity of PCR detection of A. salmonicida directly from tissues was less than 10 CFU/mg. Furthermore, a detection level of 5 fg, equivalent to approximately 1 cell, was obtained by using purified chromosomal DNA as the template. This highly reproducible assay, which requires 45 min to complete, is therefore sensitive enough to be used as a noninvasive method for monitoring fish populations for the presence of carrier fish. Because the surface protein array (A-layer) is a virulence factor of A. salmonicida, PCR analysis with oligonucleotide primers directed at vapA can also be used to provide information on the potential virulence of a strain.