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.

Detecção de prováveis deformações neotectônicas no Vale do Rio do Peixe, Região Ocidental Paulista, mediante aplicaçao de Índices RDE (Relac̈ão Declividade-Extensão) em segmentos de drenagem

Running water is one of the most important of all the physical processes which fashion the landscape, allowing gravity to operate along the valley floors. Besides this, the streams show a fast adjustment to the crustal deformations, even to the most gentle ones. This geologic behavior turns them a potential tool for neotectonic studies, specially the analysis of morphotnetric parameters associated with hydraulic gradient and discharge, this second factor being directly proportional to the extension of the streams. Both elements, gradient and stream length, can be combined in the SL index. The purpose of this paper is to show the RDE index application in the neotectonics analysis of the Rio do Peixe hydrographic basin and to compare the obtained values with the geologic basement incised by the streams. This basement encompasses Cretaceous sedimentary rocks of post-Serra Oeral Formation magmatism (Caiuá and Bauru groups) and Quaternary deposits that include chiefly recent alluvial plains and some Pleistocene terrace deposits. In the final part of this paper, an attempt is made in order to correlate the RDE results and the neotectonic framework admitted to this portion of the São Paulo State territory, as well as with field geologic, seismologic and paleoseismologic known elements. The results indicate the presence of two groups of anomalies: The first set corresponds to the Marília-Exaporã Plateau border, and the second one, located in the central portion of the hydrographic basin, is correlated to the Presidente Prudente seimogenic zone.

Análise de séries temporais de coordenadas estimadas com gps: Uma proposta metodológica para detecção, remoção e recuperação de efeitos sazonais

GPS active networks are more and more used in geodetic surveying and scientific experiments, as water vapor monitoring in the atmosphere and lithosphere plate movement. Among the methods of GPS positioning, Precise Point Positioning (PPP) has provided very good results. A characteristic of PPP is related to the modeling and / or estimation of the errors involved in this method. The accuracy obtained for the coordinates can reach few millimeters. Seasonal effects can affect such accuracy if they are not consistent treated during the data processing. Coordinates time series analyses have been realized using Fourier or Harmonics spectral analyses, wavelets, least squares estimation among others. An approach is presented in this paper aiming to investigate the seasonal effects included in the stations coordinates time series. Experiments were carried out using data from stations Manaus (NAUS) and Fortaleza (BRFT) which belong to the Brazilian Continuous GPS Network (RBMC). The coordinates of these stations were estimated daily using PPP and were analyzed through wavelets for identification of the periods of the seasonal effects (annual and semi-annual) in each time series. These effects were removed by means of a filtering process applied in the series via the least squares adjustment (LSQ) of a periodic function. The results showed that the combination of these two mathematical tools, wavelets and LSQ, is an interesting and efficient technique for removal of seasonal effects in time series.

Doses de Nitrogênio na Produção de rabanete fertirrigado e determinação de clorofila por medidor portátil nas folhas

The relative chlorophyll determination is used to predict the need for nitrogen fertilization aiming to increase production in various cultures. The objective of this study was to evaluate the soil nitrogen dose response added to the soil via fertigation in radish production and the relation between chlorophyll and cultivar Redondo Vermelho leaf nitrogen content. Transverse diameter of root, leaf area, green index, leaf N contents, shoots (stem) production, number of commercial and noncommercial roots, and the total commercial mass roots were evaluated. The N doses didn't interfere in the radish production and the readings taken with portable chlorophyll meter are not very accurate in ascertaining the level of N on radish plant growth.

Extração de vias combinando métodos de detecção de regiões e linhas em imagens de intensidade de pulso laser

This paper aims at extracting street centerlines from previously isolated street regions by using the image of laser scanning intensity. In this image, streets are easily identified, since they manifest as dark, elongate ribbons contrasting with background objects. The intensity image is segmented by using the region growing technique, which generates regions representing the streets. From these regions, the street centerlines are extracted in two manners. The first one is through the Steger lines detection method combined with a line length thresholding by which lines being shorter than a minimum length are removed. The other manner is by combining the skeletonization method of regions based on the Medial Axis Transform and with a pruning process to eliminate as much as possible the ramifications. Experiments showed that the Steger-based method provided results better than the method based on skeletonization.

Detecção de rodovias em imagens digitais de alta resolução com o uso da teoria de morfologia matemática

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detecção de áreas de oclusão para geração de ortoimagem verdadeira utilizando dados LASER

Pós-graduação em Ciências Cartográficas - FCT

Algoritmos de detecção de bordas implementados em FPGA

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Extração semi-automática de edificações com análise do modelo numérico de elevações

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

Desenvolvimento de método para determinação de acetaldeído em álcool etílico hidratado combustível por cromatografia líquida de alta eficiência com detecção eletroquímica

Pós-graduação em Química - IQ

Implementação em FPGA de um sistema para processamento de imagens digitais para aplicações diversificadas

Pós-graduação em Ciência da Computação - IBILCE

Sistema de controle de escoamento de gás natural em dutos de distribuição com detecção de vazamentos utilizando redes neurais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Medidas de deslocamento e vibrações através de imagens de câmeras digitais

Pós-graduação em Engenharia Mecânica - FEIS

Imputation of non-genotyped individuals using genotyped progeny in Nellore, a Bos indicus cattle breed

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Detecção automática e análise temporal de slope streaks na superfície de Marte

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)