990 resultados para 2-COMPONENT SYSTEM


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The decomposition of organic hydroperoxides into peroxyl radicals is a potential source of singlet molecular oxygen [O(2) ((1)Delta(g))] in biological systems. This study shows that 5-(hydroperoxymethyl)uracil (5-HPMU), a thymine hydroperoxide within DNA, reacts with metal ions or HOCl, generating O(2) ((1)Delta(g)). Spectroscopic evidence for generation of O(2) ((1)Delta(g)) was obtained by measuring (i) the bimolecular decay, (ii) the monomolecular decay, and (iii) the observation of D(2)O enhancement of O(2) ((1)Delta(g)) production and the quenching effect of NaN(3). Moreover, the presence of O(2) ((1)Delta(g)) was unequivocally demonstrated by the direct characterization of the near-infrared light emission. For the sake of comparison, O(2) ((1)Delta(g)) derived from the H(2)O(2)/HOCl system and from the thermolysis of the N,N`-di(2,3-dihydroxypropyl)-1,4-naphthalenedipropanamide endoperoxide was also monitored. More evidence of O(2) ((1)Delta(g)) generation was obtained by chemical trapping of O(2) ((1)Delta(g)) with anthracene-9,10-divinylsulfonate (AVS) and detection of the specific AVS endoperoxide by HPLC/MS/MS. The detection by HPLC/MS of 5-(hydroxymethyl)uracil and 5-formyluracil, two thymine oxidation products generated from the reaction of 5-HPMU and Ce(4+) ions, supports the Russell mechanism. These photoemission properties and chemical trapping clearly demonstrate that the decomposition of 5-HPMU generates O(2) ((1)Delta(g)) by the Russell mechanism and point to the involvement of O(2) ((1)Delta(g)) in thymidine hydroperoxide cytotoxicity. (C) 2009 Elsevier Inc. All rights reserved.

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RpfG is a paradigm for a class of widespread bacterial two-component regulators with a CheY-like receiver domain attached to a histidine-aspartic acid-glycine-tyrosine-proline (HD-GYP) cyclic di-GMP phosphodiesterase domain. In the plant pathogen Xanthomonas campestris pv. campestris (Xcc), a two-component system comprising RpfG and the complex sensor kinase RpfC is implicated in sensing and responding to the diffusible signaling factor (DSF), which is essential for cell-cell signaling. RpfF is involved in synthesizing DSF, and mutations of rpfF, rpfG, or rpfC lead to a coordinate reduction in the synthesis of virulence factors such as extracellular enzymes, biofilm structure, and motility. Using yeast two-hybrid analysis and fluorescence resonance energy transfer experiments in Xcc, we show that the physical interaction of RpfG with two proteins with diguanylate cyclase (GGDEF) domains controls a subset of RpfG-regulated virulence functions. RpfG interactions were abolished by alanine substitutions of the three residues of the conserved GYP motif in the HD-GYP domain. Changing the GYP motif or deletion of the two GGDEF-domain proteins reduced Xcc motility but not the synthesis of extracellular enzymes or biofilm formation. RpfG-GGDEF interactions are dynamic and depend on DSF signaling, being reduced in the rpfF mutant but restored by DSF addition. The results are consistent with a model in which DSF signal transduction controlling motility depends on a highly regulated, dynamic interaction of proteins that influence the localized expression of cyclic di-GMP.

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The opportunistic pathogen Pseudomonas aeruginosa PA14 possesses four fimbrial cup clusters, which may confer the ability to adapt to different environments. cupD lies in the pathogenicity island PAPI-1 next to genes coding for a putative phosphorelay system composed of the hybrid histidine kinase RcsC and the response regulator RcsB. The main focus of this work was the regulation of cupD at the mRNA level. It was found that the HN-S-like protein MvaT does not exert a strong influence on cupD transcript levels, as it does for cupA. cupD transcription is higher in cultures grown at 28 degrees C, which agrees with a cupD mutant presenting attenuated virulence only in a plant model, but not in a mouse model of infection. Whereas an rcsC in-frame deletion mutant presented higher levels of cupD mRNA, rcsB deletion had the opposite effect. Accordingly, overexpression of RcsB increased the levels of cupD transcription, and promoted biofilm formation and the appearance of fimbriae. A single transcription start site was determined for cupD and transcription from this site was induced by RcsB. A motif similar to the enterobacterial RcsB/RcsA-binding site was detected adjacent to the -35 region, suggesting that this could be the RcsB-binding site. Comparison of P. aeruginosa and Escherichia coli Rcs may provide insights into how similar systems can be used by different bacteria to control gene expression and to adapt to various environmental conditions.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Objectives: The transmission of light through translucent posts was observed, and the microhardness of light-cured cement used to secure these posts was evaluated at different depths. Methods: Fifteen single-rooted standard bovine teeth, 16 mm in size, were used. The root canals were prepared using #3 drills Light-Post (five teeth) and Aestheti Post (five teeth) systems (BISCO), with a working-length of 12 mm. In five teeth, translucent posts were cemented (Light-Post #2), while another five teeth received opaque posts (Aestheti Post #2). The roots were painted with black nail varnish to prevent the passage of light through the lateral walls of the roots. The root canals of all the specimens were treated with the All-Bond 2 adhesive system (BISCO) and cemented with light-cured cement (Enforce, Dentsply). All the roots were transversally cut to obtain six specimens 1.5 mm thick. Every two sections corresponded to a specific region of the root (cervical, middle, apical), making it possible to observe the cement microhardness at different levels. The groups (n=10) were defined as: G1: translucent post (TP)/cervical region; G2: TP/middle region; G3: TP/apical region; G4: Opaque post (OP)/cervical region; G5: OP/middle region; G6: PO/apical region. Five root canals were only filled with cement for use as a control (G7). Then, Vickers microhardness analyses were performed. Results: In G3, G5 and G6, the cement was not sufficiently hard to allow for microhardness analysis. When submitted to the ANOVA test, G1 (35.07), G2 (24.28) and G4 (28.64) presented no statistical differences. When the previous groups were compared to G7 (51.00) using the Kruskal-Wallis test, a statistical difference was found. Conclusion: Translucent posts allow cement polymerization up to the middle portion of the root.

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Purpose: To evaluate the bond strength of indirect restorations to dentin using self-adhesive cements with and without the application of adhesive systems.Material and Methods: Seventy-two bovine incisors were used, in which the buccal surfaces were ground down to expose an area of dentin measuring a minimum of 4 x 4 mm. The indirect resin composite Resilab was used to make 72 blocks, which were cemented onto the dentin surface of the teeth and divided into 4 groups (n = 18): group 1: self-adhesive resin cement BiFix SE, applied according to manufacturer's recommendations; group 2: self-adhesive resin cement RelyX Unicem, used according to manufacturer's recommendations; group 3: etch-and-rinse Solobond M adhesive system + BiFix SE; group 4: etch-and-rinse Single Bond 2 adhesive system + RelyX Unicem. The specimens were sectioned into sticks and subjected to microtensile testing in a universal testing machine (EMIC DL-200MF). Data were subjected to one-way ANOVA and Tukey's test (alpha = 5%).Results: The mean values (+/- standard deviation) obtained for the groups were: group 1: 15.28 (+/- 8.17)(a), group 2: 14.60 (+/- 5.21)(a), group 3: 39.20 (+/- 9.98)(c), group 4: 27.59 (+/- 6.57)(b). Different letters indicate significant differences (ANOVA; p = 0.0000).Conclusion: The application of adhesive systems before self-adhesive cements significantly increased the bond strength to dentin. In group 2, RelyX Unicem associated with the adhesive system Single Bond 2 showed significantly lower mean tensile bond strengths than group 3 (BiFix SE associated with the etch-and-rinse Solobond M adhesive system).

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The aim of this paper is to analyse the influence of the load centre of gravity on heavy vehicle acceleration. This analysis is done through a method in which a vehicle centre of gravity map is used. A model for the driving force is presented for bus, truck and tractor-semi trailer combinations. The proposed model takes into consideration the resistance forces (drag, rolling resistance, translation and rotation acceleration, climbing resistance) and the 4 X 2 traction system. The positions of the vehicle centre of gravity as a function of the position of the load centre of gravity are determined. The vehicle acceleration is calculated based on the position of the load centre of gravity. This study analyses the acceleration of one of the Mercedes-Benz do Brasil tractor-semitrailer vehicle. A comparison of the acceleration for different maximum adhesion coefficients and ramps are presented, showing new results. An example showing the variations of the load centre of gravity position with the acceleration time and distance is provided. The load centre of gravity position is important for vehicle safety and the efficiency and economy in the transportation of the load.

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Purpose: This study tested the hypothesis that the tribochemical silica coating on ceramic surfaces increases the bond strength of resin cement to a glass-infiltrated zirconium-based ceramic. Materials and Methods: Fifteen blocks of In-Ceram Zirconia from CEREC InLab (5 per group) and 15 composite blocks (Z-250) 5 mm x 5 mm x 4 mm were made. The ceramic surfaces were polished, and the blocks were divided into three groups: (1) airborne abrasion with 110-μm aluminum oxide particles; (2) Rocatec system, tribochemical silica coating; and (3) CoJet system, tribochemical silica coating. The ceramic blocks were cemented to the composite blocks using Panavia F according to the manufacturer's specifications. All samples were stored in 37°C distilled water for 7 days and later sectioned in two axes using a diamond disk under cooling to obtain specimens with a cross-sectional area of approximately 1 mm2 (n = 45). Each specimen was then attached with cyanoacrylate glue to an adapted device for the microtensile test, which was carried out on a universal testing machine. Results: The results were subjected to ANOVA and Tukey's test. Group 2 (23.0 ± 6.7 MPa) and group 3 (26.8 ± 7.4 MPa) showed greater bond strength than group 1 (15.1 ± 5.3 MPa). There was no significant difference between groups 2 and 3. All failures were in the adhesive zone. Conclusion: The hypothesis was confirmed - the tribochemical systems increased the bond strength between Panavia F and In-Ceram Zirconia.

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Aim : To compare the push-out strength of bovine- and human-root dentin and, thus, evaluate the suitability of bovine-root dentin to substitute human-root dentin for bond strength testing. Materials and Methods : Ten single-rooted human-teeth and ten bovine incisors were prepared using a #3 bur of a fiber post system (12 mm long). The posts were duplicated with resin cement (Duolink). The root canals were treated with All Bond 2 adhesive system and the resin posts were cemented using Duolink. The specimens were cut perpendicular to their long axis, yielding disc-specimens with 1.5 mm thickness, which were submitted to a push-out test (1 mm/min). Ten bond strength values per group (n = 10) were used for statistical analysis (Student t test, a =.05). Results : Statistically significant differences were found for the bond strength values between bovine- (4.1 1.3 MPa) and human-root dentin (8.6 5.7 MPa) (P =.0001). Conclusion : The push-out strengths of bovine- and human-root dentin were statistically different.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Pós-graduação em Ciências Farmacêuticas - FCFAR

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Pós-graduação em Desenvolvimento Humano e Tecnologias - IBRC