Docking and small angle X-ray scattering studies of purine nucleoside phosphorylase

Docking simulations have been used to assess protein complexes with some success. Small angle X-ray scattering (SAXS) is a well-established technique to investigate protein spatial configuration. This work describes the integration of geometric docking with SAXS to investigate the quaternary structure of recombinant human purine nucleoside phosphorylase (PNP). This enzyme catalyzes the reversible phosphorolysis of N-ribosidic bonds of purine nucleosides and deoxynucleosides. A genetic deficiency due to mutations in the gene encoding for PNP causes gradual decrease in T-cell immunity. Inappropriate activation of T-cells has been implicated in several clinically relevant human conditions such as transplant rejection, rheumatoid arthritis, lupus, and T-cell lymphomas. PNP is therefore a target for inhibitor development aiming at T-cell immune response modulation and has been submitted to extensive structure-based drug design. The present analysis confirms the trimeric structure observed in the crystal. The potential application of the present procedure to other systems is discussed. (C) 2003 Elsevier B.V. All rights reserved.

Cloning, overexpression, and purification of functional human purine nucleoside phosphorylase

Purine nucleoside phosphorylase (PNP) catalyzes the phosphorolysis of the N-ribosidic bonds of purine nucleosides and deoxynucleosides. A genetic deficiency due to mutations in the gene encoding for human PNP causes T-cell deficiency as the major physiological defect. Inappropriate activation of T-cells has been implicated in several clinically relevant human conditions such as transplant tissue rejection, psoriasis, rheumatoid arthritis, lupus, and T-cell lymphomas. Human PNP is therefore a target for inhibitor development aiming at T-cell immune response modulation. In addition, bacterial PNP has been used as reactant in a fast and sensitive spectrophotometric method that allows both quantitation of inorganic phosphate (Pi) and continuous assay of reactions that generate P i such as those catalyzed by ATPases and GTPases. Human PNP may therefore be an important biotechnological tool for P i detection. However, low expression of human PNP in bacterial hosts, protein purification protocols involving many steps, and low protein yields represent technical obstacles to be overcome if human PNP is to be used in either high-throughput drug screening or as a reagent in an affordable P i detection method. Here, we describe PCR amplification of human PNP from a liver cDNA library, cloning, expression in Escherichia coli host, purification, and activity measurement of homogeneous enzyme. Human PNP represented approximately 42% of total soluble cell proteins with no induction being necessary to express the target protein. Enzyme activity measurements demonstrated a 707-fold increase in specific activity of cloned human PNP as compared to control. Purification of cloned human PNP was achieved by a two-step purification protocol, yielding 48 mg homogeneous enzyme from 1 L cell culture, with a specific activity value of 80 U mg -1. © 2002 Elsevier Science (USA). All rights reserved.

O contexto afetivo-familiar de relações incestuosas entre irmãos

Pós-graduação em Psicologia - FCLAS

Surface morphology of Diplodon expansus (Kuster, 1856; Mollusca, Bivalvia, Hyriidae) gill filaments after exposure to environmentally relevant concentrations of atrazine herbicide

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

Effects of culture conditions on the production of inulinase by Kluyveromyces marxianus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Determination of selenium in nutritionally relevant foods by graphite furnace atomic absorption spectrometry using arsenic as internal standard

A method has been developed for the direct determination of Se in nutritionally relevant foods by graphite furnace atomic absorption spectrometry. Tungsten/rhodium carbide coating on the integrated platform of a transversely heated graphite atomizer or W coating with co-injection of Pd(NO3)(2) were used as a permanent modifiers. Samples and reference solutions were spiked with 500 mu g L-1 As and absorbance variations due to changes in experimental conditions were minimized. For 20 mu L aqueous analytical solutions delivered into the graphite tube, analytical curves in the 5.0-40 mu g L-1 with good linear correlation were established. Pyrolysis and atomization temperatures were evaluated using pyrolysis and atomization curves, respectively. The optimized heating program (temperature, ramp time, hold time) of the graphite tube of the Perkin-Elmer SIMAA 6000 atomic absorption spectrometer was: dry steps (110 degrees C, 5 s, 10 s; 130 degrees C, 15 s, 15 s); air-assisted pyrolysis step (600 degrees C, 20 s, 40 s; 20 degrees C, 1 s, 40 s); pyrolysis step (1300 degrees C, 10 s, 20 s); atomization step (2100 degrees C, 0 s, 4 s); clean step (2550 degrees C, 1 s, 5 s). The method was applied for Se determination in coconut water, coconut milk, soybean milk, cow milk, tomato juice, mango juice, grape juice and drinking water samples and four standard reference materials and results were in agreement at 95% confidence level. The lifetime of the tube was 500 firings and the relative standard deviations of measurements of typical samples containing 25 mu gL(-1) Se were 3.0% and 6.0% (n = 12) with and without internal standardization, respectively. The limits of detection were in the 0.35 mu g L-1-0.7 mu g Se L-1 range. The accuracy of the proposed method was evaluated by an addition-recovery experiment and all recovered values were in the 98-109% range. (c) 2004 Elsevier Ltd. All rights reserved.

A aprendizagem matemática em uma posição de fronteira: Foregrounds e intencionalidade de estudantes de uma favela brasileira

In a large metropolis, students from different neighbourhoods can experience very different life opportunities. This can influence their attitude towards schooling and learning, including the learning of mathematics. We interviewed a group of six students from a favela in a large city in the interior of the state of São Paulo in Brazil. We invited the students to look into their future and explore whether or not there could be learning motives that linked mathematics in school to possible out-of-school practices, either in terms of possible future jobs or further studies. We identified some themes in the students' descriptions of their experiences. The first theme is discrimination. The students feel discriminated against due to the fact that they come from a poor neighbourhood. They fear being trapped in some stereotypes. The second theme is escape. There is a strong motivation to begin a new life away from the favela. A third theme concerns the obscurity of mathematics. It seems clear to everybody that education is relevant to ensure a change in life. However, the mathematics lessons do not provide any clues regarding how mathematics might function is this respect. The fourth theme is uncertainty with respect to the future. The students could easily formulate almost unattainable aspirations, while reality might impose some very heavy limitations. In this article we introduce a theoretical framework for discussing the relation between favela students' life conditions in relation to their educational experiences and opportunities. Students' intentions for learning are related to their foregrounds, that is, how they perceive their future possibilities, as made evident to them by their social environment. Students in a favela could experience what we call a borderland position, a relational space where individuals meet their social environment and come to terms with the multiple choices that cultural and economic diversity make available to them.

Distributional potential of the Triatoma brasiliensis species complex at present and under scenarios of future climate conditions

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Lifetime comparison of Y-TZP/porcelain crowns under different loading conditions

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pre-task music improves swimming performance

The purpose of this study was to investigate the effects of pre--task music on swimming performance and other psychological variables. A randomized counterbalanced within--subjects (experimental and control condition) design was employed. Eighteen regional level male swimmers performed two 200m freestyle swimming time trials. Participants were exposed to either 5min of self--selected music (pre--task music condition) or 5min of silence (control condition) and, after 1 minute, performed the swimming task. Swimming time was significantly shorter (--1.44%) in the pre--task music condition. Listening to pre--task music increased motivation to perform the swimming task, while arousal remained unchanged. While fatigue increased after the swimming task in both conditions, vigour, ratings of perceived exertion and affective valence were unaltered. It is concluded, for the first time, that pre--task music improves swimming performance.

Influência da música preferida no desempenho e estados afetivos de idosos durante exercício de intensidade auto-selecionada

Low adherence to physical exercise programs is a factor that contributes to the high rates of physical inactivity worldwide. In this sense, strategies to improve the affective responses during exercise are used to impact adherence, as the use of music and self-selected exercise intensity. Despite the benefits of these strategies, there are few studies involving the combination of both (self-selected intensity and music), especially in the elderly population. Thus, the aim of this study was to investigate the influence of music on performance, ratings of perceived exertion (RPE) and on affective and motivational responses of elderly during walking exercise with self-selected intensity. Eleven elderly performed walking sessions on an athletic track in three conditions: without music, with motivational music and with oudeterous music. RPE, mood states, motivation, affective responses, session RPE and attitudes related to the experience of exercise were evaluated. Results demonstrated that both motivational and oudeterous music improved affective responses and decreased RPE. However, only the motivational music condition showed an effect on distance walked, the only presenting a consistent increase in distance. Therefore, it is concluded that the use of music, particularly motivational music, besides increasing the distance walked, reduces RPE and increase pleasure of the elderly during the exercise. Such strategy, in addition to potentially impact adherence positively, can lead to greater benefits and physical adaptations related to the training in the long term