Under flow impedimetric measurements using magnetic particles for label-free detection affinity target

The necessity to adapt sensors based on electrochemical techniques for high throughput analysis control increases the interest to develop new analytical systems able to perform measurements under buffer now. In this report we explored the possibility of employing a new system to make impedimetric measurements to detect the interaction between proteins and small molecules. The well-known biotin-streptavidin interaction was adopted to evaluate the proposed assembly. This system allows us to perform experiments under flow. Magnetic beads functionalized with streptavidin were used and first characterized using AFM and FTIR. Non-faradic impedance spectroscopy allowed the detection of the biotin-streptavidin interaction. Using our new system and under a flow of PBS buffer, 5 10-5 M of biotin was detected with a stable signal. (c) 2007 Elsevier B.V. All rights reserved.

Simultaneous analysis of five antidepressant drugs using direct injection of biofluids in a capillary restricted-access media-liquid chromatography-tandem mass spectrometry system

Direct analysis, with minimal sample pretreatment, of antidepressant drugs, fluoxetine, imipramine, desipramine, amitriptyline, and nortriptyline in biofluids was developed with a total run time of 8 min. The setup consists of two HPLC pumps, injection valve, capillary RAM-ADS-C18 pre-column and a capillary analytical C 18 column connected by means of a six-port valve in backflush mode. Detection was performed with ESI-MS/MS and only 1 mu m of sample was injected. Validation was adequately carried out using FLU-d(5) as internal standard. Calibration curves were constructed under a linear range of 1-250 ng mL(-1) in plasma, being the limit of quantification (LOQ), determined as 1 ng mL(-1), for all the analytes. With the described approach it was possible to reach a quantified mass sensitivity of 0.3 pg for each analyte (equivalent to 1.1-1.3 fmol), translating to a lower sample consumption (in the order of 103 less sample than using conventional methods). (C) 2008 Elsevier B.V. All rights reserved.

Fluoxetine and norfluoxetine analysis by direct injection of human plasma in a column switching liquid chromatographic system

A column switching LC method is presented for the analysis of fluoxetine (FLU) and norfluoxetine (NFLU) by direct injection of human plasma using a lab-made restricted access media (RAM) column. A RAM-BSA-octadecyl silica (C-18) column (40 min x 4.6 mm, 10 mu m) is evaluated in both backflush and foreflush elution modes and coupled with a C-18 lab-made (50 mm x 4.6 mm, 3 pm) analytical column in order to perform online sample preparation. Direct injection of 100 mu L, of plasma samples is possible with the developed approach. In addition, reduction of sample handling is obtained when compared with traditional liquid-liquid extraction (LLE) and SPE. The total analysis time is around 20 min. A LOQ of 15 ng/mL is achieved in a concentration range of 15-500 ng/mL, allowing the therapeutic drug monitoring of clinical samples. The precision values achieved are lower than 15% for all the evaluated points with adequate recovery and accuracy. Furthermore, no matrix interferences are found in the analysis and the proposed method shows to be an adequate alternative for analysis of FLU in plasma.

A scenario-driven decision support system for serious crime investigation

Consideration of a wide range of plausible crime scenarios during any crime investigation is important to seek convincing evidence and hence to minimize the likelihood of miscarriages of justice. It is equally important for crime investigators to be able to employ effective and efficient evidence-collection strategies that are likely to produce the most conclusive information under limited available resources. An intelligent decision support system that can assist human investigators by automatically constructing plausible scenarios, and reasoning with the likely best investigating actions will clearly be very helpful in addressing these challenging problems. This paper presents a system for creating scenario spaces from given evidence, based on an integrated application of techniques for compositional modelling and Bayesian network-based evidence evaluation. Methods of analysis are also provided by the use of entropy to exploit the synthesized scenario spaces in order to prioritize investigating actions and hypotheses. These theoretical developments are illustrated by realistic examples of serious crime investigation.

Involvement of free radicals in peroxidatic reactions catalyzed by chloroperoxidase

The mechanism of chloroperoxidase (CPO)-catalyzed peroxidatic reactions of several substituted hydroquinones was studied at various hydrogen peroxide concentrations. The pathway was studied using cytochrome c as the radical trapping agent. As the hydroquinones became more hindered there was a difference in the amount of radicals trapped. For hydroquinone, 59.3% radical pathway, and methylhydroquinone, 81.4% radical, the difference in radicals trapped is due to a difference in pathway. For 2,3-dimethylhydroquinone (75.4%), trimethylhydroquinone (44.5%), and t-butylhydroquinone (0%) other non-peroxidatic reactions are noticed. Thus, for the more substituted hydroquinones the difference in radicals trapped can not be assigned to a difference in radical pathway. Also, there were problems drawing conclusions for this system due to the catalytic reaction of hydrogen peroxide. The radical trapping ability of 2,4,6-trimethylphenol was investigated for various other substrates. TMP reacted with the radicals generated in the enzymatic reactions of phenol, resorcinol, and m-methoxyphenol. Thus, this TMP system offers further potential as another radical trapping agent for use in these studies.

Flood Risk Management In Small Urban River Using A Sustainable Urban Drainage System: Wortley Beck, Leeds, Uk

In the UK, urban river basins are particularly vulnerable to flash floods due to short and intense rainfall. This paper presents potential flood resilience approaches for the highly urbanised Wortley Beck river basin, south west of the Leeds city centre. The reach of Wortley Beck is approximately 6km long with contributing catchment area of 30km2 that drain into the River Aire. Lower Wortley has experienced regular flooding over the last few years from a range of sources, including Wortley Beck and surface and ground water, that affects properties both upstream and downstream of Farnley Lake as well as Wortley Ring Road. This has serious implications for society, the environment and economy activity in the City of Leeds. The first stage of the study involves systematically incorporating Wortley Beck’s land scape features on an Arc-GIS platform to identify existing green features in the region. This process also enables the exploration of potential blue green features: green spaces, green roofs, water retention ponds and swales at appropriate locations and connect them with existing green corridors to maximize their productivity. The next stage is involved in developing a detailed 2D urban flood inundation model for the Wortley Beck region using the CityCat model. CityCat is capable to model the effects of permeable/impermeable ground surfaces and buildings/roofs to generate flood depth and velocity maps at 1m caused by design storm events. The final stage of the study is involved in simulation of range of rainfall and flood event scenarios through CityCat model with different blue green features. Installation of other hard engineering individual property protection measures through water butts and flood walls are also incorporated in the CityCat model. This enables an integrated sustainable flood resilience strategy for this region.

A low cost one-camera optical tracking system for indoor wide-area augmented and virtual reality environments

In the last years the number of industrial applications for Augmented Reality (AR) and Virtual Reality (VR) environments has significantly increased. Optical tracking systems are an important component of AR/VR environments. In this work, a low cost optical tracking system with adequate attributes for professional use is proposed. The system works in infrared spectral region to reduce optical noise. A highspeed camera, equipped with daylight blocking filter and infrared flash strobes, transfers uncompressed grayscale images to a regular PC, where image pre-processing software and the PTrack tracking algorithm recognize a set of retro-reflective markers and extract its 3D position and orientation. Included in this work is a comprehensive research on image pre-processing and tracking algorithms. A testbed was built to perform accuracy and precision tests. Results show that the system reaches accuracy and precision levels slightly worse than but still comparable to professional systems. Due to its modularity, the system can be expanded by using several one-camera tracking modules linked by a sensor fusion algorithm, in order to obtain a larger working range. A setup with two modules was built and tested, resulting in performance similar to the stand-alone configuration.

Contratos de aliança e formas híbridas de contratos de construção

Este trabalho analisa a sistemática de contratações inerentes à atividade de construção civil, inserida num ambiente de liberdade de contratação e levando em consideração a criação de modelos atípicos e, portanto, híbridos. Para tal, faz-se necessária uma gama de subsídios: aplicação de argumentos econômicos; capacidade de operacionalizar objetivamente as demandas oriundas de tais modalidades de contratos, que sempre visa maximizar seus resultados; necessidade de velocidade e economia de tempo nas construções. Há de se destacar que os referidos modelos de contrato são híbridos, sim, mas nem por isso ilícitos, posto que, no direito brasileiro, vigora o princípio de liberdade de contratar, desde que tais contratações estejam assentadas em princípios básicos estabelecidos por lei, quais sejam: objeto lícito, agentes capazes e atendimento à forma prescrita ou não defesa em lei. Esse ambiente de liberdade de contratação propiciou a criação de modelos atípicos e moventes de contratos de construção, que variam e se mesclam entre si sob os mais diversos aspectos, com destaque para: alocação de riscos; forma de remuneração e aplicação de penalidades, o que impacta diretamente na forma, conteúdo e valor do orçamento a ser apresentado. Desse modo, é imperioso que as rédeas desses diversos modelos de contratação estejam direcionadas pelo racional econômico, não só em grandes obras de infraestrutura, mas também em outros empreendimentos, fazendo emergir uma cultura de participação entre os contratantes, de forma a propiciar obras mais rápidas e com um menor custo, que seria de fundamental importância para o desenvolvimento do país.

A new and improved strategy combining a dispersive-solid phase extraction-based multiclass method with ultra high pressure liquid chromatography for analysis of low molecular weight polyphenols in vegetables

This paper reports on the development and optimization of a modified Quick, Easy, Cheap Effective, Rugged and Safe (QuEChERS) based extraction technique coupled with a clean-up dispersive-solid phase extraction (dSPE) as a new, reliable and powerful strategy to enhance the extraction efficiency of free low molecular-weight polyphenols in selected species of dietary vegetables. The process involves two simple steps. First, the homogenized samples are extracted and partitioned using an organic solvent and salt solution. Then, the supernatant is further extracted and cleaned using a dSPE technique. Final clear extracts of vegetables were concentrated under vacuum to near dryness and taken up into initial mobile phase (0.1% formic acid and 20% methanol). The separation and quantification of free low molecular weight polyphenols from the vegetable extracts was achieved by ultrahigh pressure liquid chromatography (UHPLC) equipped with a phodiode array (PDA) detection system and a Trifunctional High Strength Silica capillary analytical column (HSS T3), specially designed for polar compounds. The performance of the method was assessed by studying the selectivity, linear dynamic range, the limit of detection (LOD) and limit of quantification (LOQ), precision, trueness, and matrix effects. The validation parameters of the method showed satisfactory figures of merit. Good linearity (View the MathML sourceRvalues2>0.954; (+)-catechin in carrot samples) was achieved at the studied concentration range. Reproducibility was better than 3%. Consistent recoveries of polyphenols ranging from 78.4 to 99.9% were observed when all target vegetable samples were spiked at two concentration levels, with relative standard deviations (RSDs, n = 5) lower than 2.9%. The LODs and the LOQs ranged from 0.005 μg mL−1 (trans-resveratrol, carrot) to 0.62 μg mL−1 (syringic acid, garlic) and from 0.016 μg mL−1 (trans-resveratrol, carrot) to 0.87 μg mL−1 ((+)-catechin, carrot) depending on the compound. The method was applied for studying the occurrence of free low molecular weight polyphenols in eight selected dietary vegetables (broccoli, tomato, carrot, garlic, onion, red pepper, green pepper and beetroot), providing a valuable and promising tool for food quality evaluation.

Applying DEMO to model and automate a process in a workflow system: a case study in a city hall

Organizations are Complex systems. A conceptual model of the enterprise is needed that is: coherent the distinguished aspect models constitute a logical and truly integral comprehensive all relevant issues are covered consistent the aspect models are free from contradictions or irregularities concise no superfluous matters are contained in it essential it shows only the essence of the enterprise, i.e., the model abstracts from all realization and implementation issues. The world is in great need for transparency about the operation of all the systems we daily work with, ranging from the domestic appliances to the big societal institutions. In this context the field of enterprise ontology has emerged with the aim to create models that help to understand the essence of the construction and operation of complete systems; more specifically, of enterprises. Enterprise ontology arises in the way to look through the distracting and confusing appearance of an enterprise right into its deep kernel. This, from the perspective of the system designer gives him the tools needed to design a successful system in a way that’s reflects the desires and needs of the workers of the enterprise. This project’s context is the use of DEMO (Design and Engineering Methodology for Organizations) for (re)designing or (re)engineering of an enterprise, namely a process of the construction department of a city hall, the lack of a well-founded theory about the construction and operation of this processes that was the motivation behind this work. The purpose of studying applying the DEMO theory and method was to optimize the process, automating it as much as possible, while reducing paper and time spent between tasks and provide a better service to the citizens.

Is the meander growth in the Brazil Current system off Southeast Brazil due to baroclinic instability?

Temporally-growing frontal meandering and occasional eddy-shedding is observed in the Brazil Current (BC) as it flows adjacent to the Brazilian Coast. No study of the dynamics of this phenomenon has been conducted to date in the region between 22 degrees S and 25 degrees S. Within this latitude range, the flow over the intermediate continental slope is marked by a current inversion at a depth that is associated with the Intermediate Western Boundary Current (IWBC). A time series analysis of 10-current-meter mooring data was used to describe a mean vertical profile for the BC-IWBC jet and a typical meander vertical structure. The latter was obtained by an empirical orthogonal function (EOF) analysis that showed a single mode explaining 82% of the total variance. This mode structure decayed sharply with depth, revealing that the meandering is much more vigorous within the BC domain than it is in the IWBC region. As the spectral analysis of the mode amplitude time series revealed no significant periods, we searched for dominant wavelengths. This search was done via a spatial EOF analysis on 51 thermal front patterns derived from digitized AVHRR images. Four modes were statistically significant at the 95% confidence level. Modes 3 and 4, which together explained 18% of the total variance, are associated with 266 and 338-km vorticity waves, respectively. With this new information derived from the data, the [Johns, W.E., 1988. One-dimensional baroclinically unstable waves on the Gulf Stream potential vorticity gradient near Cape Hatteras. Dyn. Atmos. Oceans 11, 323-350] one-dimensional quasi-geostrophic model was applied to the interpolated mean BC-IWBC jet. The results indicated that the BC system is indeed baroclinically unstable and that the wavelengths depicted in the thermal front analysis are associated with the most unstable waves produced by the model. Growth rates were about 0.06 (0.05) days(-1) for the 266-km (338-km) wave. Moreover, phase speeds for these waves were low compared to the surface BC velocity and may account for remarks in the literature about growing standing or stationary meanders off southeast Brazil. The theoretical vertical structure modes associated with these waves resembled very closely to the one obtained for the current-meter mooring EOF analysis. We interpret this agreement as a confirmation that baroclinic instability is an important mechanism in meander growth in the BC system. (C) 2008 Elsevier B.V. All rights reserved.

Weed interference on maize (Zea mays) under no tillage system

A field trial was carried out in Brazil in March 2002 with the aim to evaluate the effects of different timing and extension of weedy period on maize productivity. The hybrid Pioneer 30K75 was sowed under 7 t ha(-1) mulching promoted by glyphosate spraying. The treatments were divided in two groups: In the first group, weeds were maintained since the maize sowing until different periods in the crop cycle: 0, 14, 28, 42, 56, 70, and 150 days (harvesting time). In the second group, the maize crop was kept weed free for the same periods of the first group. Weed control was done through hand hoeing. A complete randomized blocks experimental design with five replications was used for plots distribution in the field. Nonlinear regression model was used to study the effects of weedy or weedfree periods on maize productivity. Weed community included 13 families and 31 species. Asteraceae, Poaceae, and Euphorbiaceae were the most abundant families. Results showed that under no tillage condition with 7 t ha-1 mulching at sowing time, the maize crop could cohabit with weed community for 54 days without any yield lost. on the other hand, if the crop was kept weed free for 27 days, the weed interference was not enable to reduce maize production. According to these results one weed control measure between 27 and 54 days after crop emergence could be enough to avoid any reduction in maize productivity.

Campo educacional e representação social da formação docente :o olhar dos agentes

Teaching formation has been the target of many changes, having been forged according to numerous formats and models through different times and spaces, composing thus, different codes and curricular proposals for different levels of qualification. We've tried in this work to pinpoint the main discussions which take place in the area of the teaching formation, based in the views of modern writers. We ve also tried, through the Social Representations Theory, to reveal and learn how the agents directly involved in this process realize and react, that is, the teachers who cope with the beginning stages of the Fundamental Teaching in the public school system in Natal/RN, taking into account that, in many cases, such formation demands the rethinking of the very formation policies. We've also adopted the concepts of field and educational field introduced by the praxiology of Pierre Bourdieu. It s been considered a fundamental theoretical reference which enables the understanding of social phenomena, both in macro and micro viewpoints. Thus, we do not neglect the whole, and particularly, the nuances of each context or specific situation. In the methodological track we ve used for data gathering the Free Word Association Test, and the Semi-Structured Interview, and also secondary sources for the characterization of the research spectre. Data treatment and analysis were performed with the help of the following software: SPSS Statistical Package for Social Sciences and EVOC; and the method of Content Category Analysis. The combination of the concepts and techniques mentioned above was necessary to cope with the qualitative and quantitative aspects, in our attempt to offer a wider range of contributions and outcome validations, which have shown, among other less explicit elements, the existence of a social representation of the teaching formation such as: knowledge theoretical and practical; a necessity imposed by the symbolical conflicts of the social field; capacitation and compromisse. We acknowledge the relevance of the thoughts discussed here, though aware that this is just one of the possible approaches to the theme

Construction of an alkaline phosphatase-liposome system: a tool for biomineralization study

Alkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper. we standardize a method for construction an alkaline phosphatase liposome system to mimic matrix vesicles and examine a some kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into liposomes constituted from dimyristoylphosphatidylcholine (DMPC), dilaurilphosphatidylcholine (DLPC) or dipalmitoylphosphatidylcholine (DPPC). This process was time-dependent and >95% of the enzyme was incorporated into the liposome after 4 h of incubation at 25 degreesC. Although, incorporation was more rapid when vesicles constituted from DPPC were used, the incorporation was more efficient using vesicles constituted from DMPC. The 395 nm diameter of the alkaline phosphatase-liposome system was relatively homogeneous and more stable when stored at 4 degreesC.Alkaline phosphatase was completely released from liposome system only using purified phosphatidylinositol-specific phospholipase C (PIPLC). These experiments confirm that the interaction between alkaline phosphatase and lipid bilayer of liposome is via GPI anchor of the enzyme, alone. An important point shown is that an enzyme bound to liposome does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but a liposome environment affects its kinetic properties, specifically for pyrophosphate.The standardization of such system allows the study of the effect of phospholipids and the enzyme in in vitro and in vivo mineralization, since it reproduces many essential features of the matrix vesicle. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Erythrocyte ghost cell-alkaline phosphatase: construction and characterization of a vesicular system for use in biomineralization studies

Alkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper we standardize a method to construction a resealed ghost cell-alkaline phosphatase system to mimic matrix vesicles and examine the kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into resealed ghost cells. This process was time-dependent and practically 50% of the enzyme was incorporated into the vesicles in 40 h of incubation, at 25 degreesC. Alkaline phosphatase-ghost cell systems were relatively homogeneous with diameters of about 300 nm and were more stable when stored at -20 degreesC.Alkaline phosphatase was completely released from the resealed ghost cell-system using only phospholipase C. These experiments confirm that the interaction between alkaline phosphatase and the lipid bilayer of resealed ghost cell is exclusively via glycosylphosphatidylinositol (GPI) anchor of the enzyme.An important point shown is that an enzyme bound to resealed ghost cell does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but the presence of a ghost membrane, as a support of the enzyme, affects its kinetic properties. Moreover, calcium ions stimulate and phosphate ions inhibit the PNPPase activity of alkaline phosphatase present in resealed ghost cells. (C) 2002 Elsevier B.V. B.V. All rights reserved.