Characterisation of the intracellular-glutamate decarboxylase system (GAD): analysis of its function, transcription and role in the acid resistance of various strains of Listeria monocytogenes

The glutamate decarboxylase (GAD) system is important for the acid resistance of Listeria monocytogenes. We previously showed that under acidic conditions, glutamate (Glt)/γ-aminobutyrate (GABA) antiport is impaired in minimal media but not in rich ones, like brain heart infusion. Here we demonstrate that this behavior is more complex and it is subject to strain and medium variation. Despite the impaired Glt/GABA antiport, cells accumulate intracellular GABA (GABA(i)) as a standard response against acid in any medium, and this occurs in all strains tested. Since these systems can occur independently of one another, we refer to them as the extracellular (GAD(e)) and intracellular (GAD(i)) systems. We show here that GAD(i) contributes to acid resistance since in a ΔgadD1D2 mutant, reduced GABA(i) accumulation coincided with a 3.2-log-unit reduction in survival at pH 3.0 compared to that of wild-type strain LO28. Among 20 different strains, the GAD(i) system was found to remove 23.11% ± 18.87% of the protons removed by the overall GAD system. Furthermore, the GAD(i) system is activated at milder pH values (4.5 to 5.0) than the GAD(e) system (pH 4.0 to 4.5), suggesting that GAD(i) is the more responsive of the two and the first line of defense against acid. Through functional genomics, we found a major role for GadD2 in the function of GAD(i), while that of GadD1 was minor. Furthermore, the transcription of the gad genes in three common reference strains (10403S, LO28, and EGD-e) during an acid challenge correlated well with their relative acid sensitivity. No transcriptional upregulation of the gadT2D2 operon, which is the most important component of the GAD system, was observed, while gadD3 transcription was the highest among all gad genes in all strains. In this study, we present a revised model for the function of the GAD system and highlight the important role of GAD(i) in the acid resistance of L. monocytogenes.

Evaluation of Postpolymerization as a Function of the Storage Time of Triethylene Glycol Dimethacrylate/2,2-Bis[4-(2-Hydroxy-3-Methacryloxy-Prop-1-Oxy)-Phenyl]-propane Bisphenyl-alpha-Glycidyl Ether Dimethacrylate Copolymers Used in Dental Resins by Differential Scanning Calorimetry and Dynamic Mechanical Analysis

The aim of this work was to evaluate the effect of the storage time on the thermal properties of triethylene glycol dimethacrylate/2,2-bis[4-(2-hydroxy-3-methacryloxy-prop-1-oxy)-phenyl]propane bisphenyl-alpha-glycidyl ether dimethacrylate (TB) copolymers used in formulations of dental resins after photopolymerization. The TB copolymers were prepared by photopolymerization with an Ultrablue IS light-emitting diode, stored in the dark for 160 days at 37 degrees C, and characterized with differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), and Fourier transform infrared spectroscopy with attenuated total reflection. DSC curves indicated the presence of an exothermic peak, confirming that the reaction was not completed during the photopolymerization process. This exothermic peak became smaller as a function of the storage time and was shifted at higher temperatures. In DMA studies, a plot of the loss tangent versus the temperature initially showed the presence of two well-defined peaks. The presence of both peaks confirmed the presence of residual monomers that were not converted during the photopolymerization process. (C) 2009 Wiley Periodicals, Inc. J Appl Polym Sci 112: 679-684, 2009

Aggregate vs disaggregate data analysis: a paradox in the estimation of money demand function of Japan under the low interest rate policy

We investigate the issue of whether there was a stable money demand function for Japan in 1990's using both aggregate and disaggregate time series data. The aggregate data appears to support the contention that there was no stable money demand function. The disaggregate data shows that there was a stable money demand function. Neither was there any indication of the presence of liquidity trapo Possible sources of discrepancy are explored and the diametrically opposite results between the aggregate and disaggregate analysis are attributed to the neglected heterogeneity among micro units. We also conduct simulation analysis to show that when heterogeneity among micro units is present. The prediction of aggregate outcomes, using aggregate data is less accurate than the prediction based on micro equations. Moreover. policy evaluation based on aggregate data can be grossly misleading.

SMase II, a new sphingomyelinase D from Loxosceles laeta venom gland: Molecular cloning, expression, function and structural analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

FTY720 in combination with cyclosporine - an analysis of skin allograft survival and renal function

Acute and chronic nephrotoxicity caused by CsA Continuous administration impair kidney allograft survival. Several clinical and experimental protocols have shown benefits to the kidney after decreasing CsA dose, withdrawing the drug or delaying its introduction after transplantation.FTY720 is a new Compound that has immunosuppressive characteristics and increase allograft survival in animal models without causing the side effects of calcineurin inhibitors (CNIs). FTY720 described mechanism of action that consists to alter the lymphocyte migration pattern without impairment of the immune system response against pathogens.In our mice model, FTY720 administered alone or in combination with CsA during 21 days increased skin allograft survival in a fully mismatched strain combination and did not cause significant changes in renal function. Moreover, renal structure was normal in all groups suggesting that at low doses (10 mg/kg/day) CsA can be associated during short-term period to other immunosuppressive drugs, i.e. FTY720 without affecting the kidney.Combination of immunosuppressive compounds with FTY720 and/or delayed introduction of low cyclosporine dose Could prevent graft rejection and avoid nephrotoxicity. (c) 2006 Elsevier B.V. All rights reserved.

Analysis of the Paracoccidioides brasiliensis triosephosphate isomerase suggests the potential for adhesin function

Paracoccidioides brasiliensis is an important fungal pathogen. The disease it causes, paracoccidioidomycosis (PCM), ranges from localized pulmonary infection to systemic processes that endanger the life of the patient. Paracoccidioides brasiliensis adhesion to host tissues contributes to its virulence, but we know relatively little about molecules and the molecular mechanisms governing fungal adhesion to mammalian cells. Triosephosphate isomerase (TPI: EC 5.3.1.1) of P. brasiliensis (PbTPI) is a fungal antigen characterized by microsequencing of peptides. The protein, which is predominantly expressed in the yeast parasitic phase, localizes at the cell wall and in the cytoplasmic compartment. TPI and the respective polyclonal antibody produced against this protein inhibited the interaction of P. brasiliensis to in vitro cultured epithelial cells. TPI binds preferentially to laminin, as determined by peptide inhibition assays. Collectively, these results suggest that TPI is required for interactions between P. brasiliensis and extracellular matrix molecules such as laminin and that this interaction may play an important role in the fungal adherence and invasion of host cells.

Analysis of camphorquinone in composite resins as a function of shade

Objective. To identify and quantify the camphorquinone (CQ) used in different brands of composite resins as a function of the shade analyzed.Materials and methods. Filtek Z250 A3 (FZA3), Filtek Z-250 Incisal (FZI), Pyramid Enamel A1 (PEA1), Pyramid Enamel Translucent (PET), Filtek Supreme A3E (FSA3) and Filtek Supreme GT (FSGT) were used. Five hundred milligrams of each resin were weighed and then dissolved in 1.0 ml of methanol. The samples were centrifuged to accelerate the sedimentation of the inorganic particles. 0.8 ml of the supernatant solution was collected with a pipette and assessed under gas chromatography coupled to the mass spectrometer (GC-MS). The results were compared to pure CQ solutions, used as a standard. Student's t-test, (p = 0.05) significant at the level of 5%, compared the results of each brand shade.Results. A smaller amount of camphorquinone was found in Filtek Z-250 (FZI) resin incisal shade when compared to (FZA3) A3 shade. on the other hand, Filtek Supreme resin featured a statistically larger camphorquinone amount in the incisal shade. in Pyramid Enamel resin camphorquinone was found only in shade Al, while the photoinitiator used in the Translucent shade was not identified.Significance. Based on the data obtained, it is possible to conclude that a single composite resin brand may feature differences in amount and type of photoinitiator used. (C) 2006 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Three-dimensional finite element analysis of MHD duct flow by the penalty function formulation

A numerical scheme based on the Finite Element Method (FEM) is presented to calculate the full solution of a three-dimensional steady magnetohydrodynamic (MHD) flow with moderately high Hartmann numbers and interaction parameters. An incompressible, viscous and electrically conducting liquid-metal is considered. Assuming a low magnetic Reynolds number, the solution method solves the coupled Navier-Stokes and Maxwell's equations through the use of a penalty function method. Results are presented for Hartmann numbers in the range 10(2)-10(3).

Assessment of luteal function in goats by ultrasonographic image attribute analysis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Micromechanics of dentin/adhesive interface in function of dentin depth: 3d finite element analysis

Objectives: The aim of this study was to analyze the stress distribution on dentin/adhesive interface (d/a) through a 3-D finite element analysis (FEA) varying the number and diameter of the dentin tubules orifice according to dentin depth, keeping hybrid layer (HL) thickness and TAǴs length constant. Materials and Methods: 3 models were built through the SolidWorks software: SD - specimen simulating superficial dentin (41 x 41 x 82 μm), with a 3 μm thick HL, a 17 μm length Tag, and 8 tubules with a 0.9 μm diameter restored with composite resin. MD - similar to M1 with 12 tubules with a 1.2 μm diameter, simulating medium dentin. DD - similar to M1 with 16 tubules with a 2.5 μm diameter, simulating deep dentin. Other two models were built in order to keep the diameter constant in 2.5 μm: MS - similar to SD with 8 tubules; and MM - similar to MD with 12 tubules. The boundary condition was applied to the base surface of each specimen. Tensile load (0.03N) was performed on the composite resin top surface. Stress field (maximum principal stress in tension - σMAX) was performed using Ansys Wokbench 10.0. Results: The peak of σMAX (MPa) were similar between SD (110) and MD (106), and higher for DD (134). The stress distribution pathway was similar for all models, starting from peritubular dentin to adhesive layer, intertubular dentin and hybrid layer. The peak of σMAX (MPa) for those structures was, respectively: 134 (DD), 56.9 (SD), 45.5 (DD), and 36.7 (MD). Conclusions: The number of dentin tubules had no influence in the σMAX at the dentin/adhesive interface. Peritubular and intertubular dentin showed higher stress with the bigger dentin tubules orifice condition. The σMAX in the hybrid layer and adhesive layer were going down from superficial dentin to deeper dentin. In a failure scenario, the hybrid layer in contact with peritubular dentin and adhesive layer is the first region for breaking the adhesion. © 2011 Nova Science Publishers, Inc.

Color degradation of acrylic resin denture teeth as a function of liquid diet: ultraviolet-visible reflection analysis

The effect of different beverages on acrylic resin denture teeth color degradation is evaluated. Ten acrylic resin denture teeth brands were evaluated: Art Plus (AP), Biolux (BX), Biotone IPN (BI), Magister (MG), Mondial 6 (MD), Premium 6 (PR), SR Vivodent PE (SR), Trilux (TR), Trubyte Biotone (TB), and Vipi Dent Plus (VP). Teeth were immersed in staining solutions (coffee, cola, and orange juice) or artificial saliva (control) (n = 6) for 1, 7, 15, or 30 days. Specimen colors were evaluated spectrophotometrically based on the Commission Internationale d'Eclairage L*a*b* system. Color differences (Delta E) were calculated between the baseline and post-staining results. Data were evaluated by analysis of variance and Tukey test (alpha = 0.05). BI (1.82 +/- 0.95) and TR (1.78 +/- 0.72) teeth exhibited the greatest Delta E values, while BX (0.88 +/- 0.43) and MD (1.09 +/- 0.44) teeth were the lowest, regardless of solution and measurement period, and were different from BI and TR teeth (P < 0.05). Cola and coffee promoted higher denture teeth color alterations than orange juice and saliva (P < 0.05). Saliva generated the lowest denture teeth color alterations. Greater immersion times caused higher denture teeth color changes. The lifespan of removable dentures and the aesthetic satisfaction of several edentulous patients may be increased with the use of stain-resistant artificial denture teeth. (C) The Authors.

Large-scale analysis of the SDSS-III DR8 photometric luminous galaxies angular correlation function

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Analysis of renal function after on and off pump coronary artery bypass grafting

Cardiopulmonary bypass (CPB) is often associated with renal dysfunction, as measured by plasma creatinine levels and hemodialysis rates. Aim. To compare creatinine clearance (CrCl), estimated with the Cockroft and Gault formula, between patients undergoing off-pump coronary artery bypass grafting (OPCAB) versus on-pump CABG (on-CAB). Material and methods. Between April 2008 and April 2009, 119 patients underwent coronary bypass graft surgery. Fifty-eight (58) of these patients underwent OPCAB while 61 had on-CAB. Creatinine clearance, plasma creatinine levels, and clinical outcome were compared between the groups. A creatinine clearance value of 50 mL/minute was accepted as the lowest limit of normal renal function. Results. There were two hospital deaths caused by sepses after pulmonary infection. Creatinine clearance (Preoperative OPCAB 73,64±33,72 x on-CAB 75,70±34,30mL/min; discharge OPCAB 75,73±35,07 x on-CAB 79,07±34,71 mL/ min; p=0,609), and creatinine levels (Preoperative OPCAB 1,04±0,38 x on-CAB 1,13±0,53 mg/dL; discharge OPCAB 1,12±0,79 x on-CAB 1,04±0,29mg/dL; p=0,407) did not show statistically inter-group differences. Conclusion. Deterioration in renal function is associated with higher rates of postoperative complications. No significant difference in CrCl could be demonstrated between the groups.

Optimizing urban traffic flow using genetic algorithm with petri net analysis as fitness function

This paper describes a new methodology adopted for urban traffic stream optimization. By using Petri net analysis as fitness function of a Genetic Algorithm, an entire urban road network is controlled in real time. With the advent of new technologies that have been published, particularly focusing on communications among vehicles and roads infrastructures, we consider that vehicles can provide their positions and their destinations to a central server so that it is able to calculate the best route for one of them. Our tests concentrate on comparisons between the proposed approach and other algorithms that are currently used for the same purpose, being possible to conclude that our algorithm optimizes traffic in a relevant manner.