Nitrate Reductase And Glutamine Synthetase Activities In S1 Endogamic Families Of The Maize Populations Sol Da Manhã Nf And Catetão

The possibility of improving nitrogen use efficiency in maize was investigated using S1 endogamic families of the populations Sol da Manhã NF and Catetão. A simple 10 × 10 lattice design was adopted and the trials carried out at the experimental field of MITLA AGRÍCOLA LTDA, in Uberlândia, State of Minas Gerais, during the 1994/95 planting season. Based on grain production figures, the three best and three worst performing S1 endogamic families were selected for this study. These were pooled to form four sub-populations denominated NFB, NFR (the best and worst families, respectively, of the Sol da Manhã NF variety), CATB and CATR (the best and worst families, respectively, of the Catetão variety). Each of these sub-populations was evaluated under greenhouse conditions. The experimental design was factorial with treatments arranged in randomized blocks. Sample replicates consisted of pots with four plants. Feeding with modified Hoagland's nutrient solution began on the seventh day after sowing. The study involved four nitrogen regimes, where varying proportions of NO3 - and NH4 + were formulated, such that the nutrient solution contained the following mixtures: 75% NO3 -: 25% NH4 +; 25% NO3 -: 75% NH4 +; 50% NO3 -: 50% NH4 + (all high N mixtures) and 5% NO3 -: 5% NH4 + (low N mixture). Twenty-five days after planting, the activities of the enzymes nitrate reductase and glutamine synthetase (transferase and synthetase assays) were determined for the leaves using the third topmost expanded leaf of the four plants in each pot. 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Porto Oscillation Code (POSC)

The Porto Oscillation Code (POSC) has been developed in 1995 and improved over the years, with the main goal of calculating linear adiabatic oscillations for models of solar-type stars. It has also been used to estimate the frequencies and eigenfunctions of stars from the pre-main sequence up to the sub-giant phase, having a mass between 0.8 and 4 solar masses. The code solves the linearised perturbation equations of adiabatic pulsations for an equilibrium model using a second order numerical integration method. The possibility of using Richardson extrapolation is implemented. Several options for the surface boundary condition can be used. In this work we briefly review the key ingredients of the calculations, namely the equations, the numerical scheme and the output.Comment: Accepted for publication in Astrophysics and Space Science

Scaling up a chemically-defined aggregate-based suspension culture system for neural commitment of human pluripotent stem cells

The demand of high cell numbers for applications in cellular therapies and drug screening requires the development of scalable platforms capable to generating highly pure populations of tissue-specific cells from human pluripotent stem cells. This work describes the scaling-up of an aggregate-based culture system for neural induction of human induced pluripotent stem cells (hiPSCs) under chemically-defined conditions. Since initial cell density and aggregate size have an important impact in the expansion and commitment of these cells into a particular lineage, a combination of non-enzymatic dissociation and rotary agitation was successfully used to produce homogeneous populations of hiPSC aggregates with an optimal (140 µm) and narrow distribution of diameters (coefficient of variation of 21.6%). Scalable neural commitment of hiPSCs as 3D aggregates was then performed in 50 mL spinner flasks, and process optimization using a factorial design approach was developed involving parameters such as agitation rate and seeding density. We were able to produce neural progenitor cell cultures, that at the end of a 6-day neural induction process contained less than 3% of Oct4-positive cells and that, after replating, retained more than 60% of Pax6-positive neural cells. Furthermore, after scalable differentiation, hiPSC-derived neural progenitors still retained their multipotent potential, being able to give rise to neuronal and glial cells. The results presented in this work should set the stage for the future generation of a clinically relevant number of human neural progenitors for transplantation and other biomedical applications using totally controlled, automated and reproducible large-scale bioreactor culture systems

Texture analysis of computed tomography images of acute ischemic stroke patients

Computed tomography (CT) images are routinely used to assess ischemic brain stroke in the acute phase. They can provide important clues about whether to treat the patient by thrombolysis with tissue plasminogen activator. However, in the acute phase, the lesions may be difficult to detect in the images using standard visual analysis. The objective of the present study was to determine if texture analysis techniques applied to CT images of stroke patients could differentiate between normal tissue and affected areas that usually go unperceived under visual analysis. We performed a pilot study in which texture analysis, based on the gray level co-occurrence matrix, was applied to the CT brain images of 5 patients and of 5 control subjects and the results were compared by discriminant analysis. Thirteen regions of interest, regarding areas that may be potentially affected by ischemic stroke, were selected for calculation of texture parameters. All regions of interest for all subjects were classified as lesional or non-lesional tissue by an expert neuroradiologist. Visual assessment of the discriminant analysis graphs showed differences in the values of texture parameters between patients and controls, and also between texture parameters for lesional and non-lesional tissue of the patients. This suggests that texture analysis can indeed be a useful tool to help neurologists in the early assessment of ischemic stroke and quantification of the extent of the affected areas.1076107

A Comparison between Dual Phase Steel and Interstitial Free Steel Due To the Springback Effect

International audienceThis is a study of the springback effect on two kinds of high strength steel, which are: dualphase and interstitial free, currently used as feedstock in the production of vehicles. The mechanical characterization of the springback effect was performed by means of a mechanical conformation test, called three-point air bending, performed by adapting it to the unconstrained cylindrical bending test. It was also evaluated the mechanical properties of the material defined by the tensile test in order to determine its tensile strength, yield strength, and elongation. Furthermore, it was performed a microstructural characterization of advanced steels by identifying and quantifying the present phases in coexistence by means of digital image processing. The results indicate that the springback effect in the dual-phase steel has the highest springback rates due to its high mechanical strength, and it causes a decrease in the aspect ratio of the grains that suffered mechanical conformation attempting to return it to its original form. On the other hand, the springback effect has the lowest rates, and the change in aspect ratio depends only on the interstitial free steel elongation capacity due to its lower mechanical strength

ZapA, a possible virulence factor from Proteus mirabilis exhibits broad protease substrate specificity

The opportunistic bacterium Proteus mirabilis secretes a metalloprotease, ZapA, considered to be one of its virulence factors due to its IgA-degrading activity. However, the substrate specificity of this enzyme has not yet been fully characterized. In the present study we used fluorescent peptides derived from bioactive peptides and the oxidized ß-chain of insulin to determine the enzyme specificity. The bradykinin- and dynorphin-derived peptides were cleaved at the single bonds Phe-Ser and Phe-Leu, with catalytic efficiencies of 291 and 13 mM/s, respectively. Besides confirming already published cleavage sites, a novel cleavage site was determined for the ß-chain of insulin (Val-Asn). Both the natural and the recombinant enzyme displayed the same broad specificity, demonstrated by the presence of hydrophobic, hydrophilic, charged and uncharged amino acid residues at the scissile bonds. Native IgA, however, was resistant to hydrolysis by ZapA.Universidade de São Paulo Instituto de Ciências Biomédicas Departamento de MicrobiologiaInstituto Butantan CEPID-FAPESP Centro de Toxinologia AplicadaUniversidade Federal de São Paulo (UNIFESP) CEPID-FAPESP Centro de Toxinologia AplicadaUniversidade Estadual Paulista CEPID-FAPESP Centro de Toxinologia AplicadaUNIFESP, CEPID-FAPESP Centro de Toxinologia AplicadaSciEL

Development of an operational substrate for ZapA, a metalloprotease secreted by the bacterium Proteus mirabilis

The protease ZapA, secreted by Proteus mirabilis, has been considered to be a virulence factor of this opportunistic bacterium. The control of its expression requires the use of an appropriate methodology, which until now has not been developed. The present study focused on the replacement of azocasein with fluorogenic substrates, and on the definition of enzyme specificity. Eight fluorogenic substrates were tested, and the peptide Abz-Ala-Phe-Arg-Ser-Ala-Ala-Gln-EDDnp was found to be the most convenient for use as an operational substrate for ZapA. A single peptide bond (Arg-Ser) was cleaved with a Km of 4.6 µM, a k cat of 1.73 s-1, and a catalytic efficiency of 376 (mM s)-1. Another good substrate for ZapA was peptide 6 (Abz-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg-Gln-EDDnp) which was cleaved at a single bond (Phe-Ser) with a Km of 13.6 µM, a k cat of 3.96 s-1 and a catalytic efficiency of 291 (mM s)-1. The properties of the amino acids flanking the scissile bonds were also evaluated, and no clear requirement for the amino acid residue at P1 was found, although the enzyme seems to have a preference for a hydrophobic residue at P2.Universidade de São PauloUniversidade Federal de São Paulo (UNIFESP)Universidade Estadual de São PauloInstituto ButantanUNIFESPSciEL