Controlled oxidative protein refolding using an ion-exchange column

Column-based refolding of complex and highly disulfide-bonded proteins simplifies protein renaturation at both preparative and process scale by integrating and automating a number of operations commonly used in dilution refolding. Bovine serum albumin (BSA) was used as a model protein for refolding and oxido-shuffling on an ion-exchange column to give a refolding yield of 55 % after 40 Ih incubation. Successful on-column refolding was conducted at protein concentrations of up to 10 mg/ml and refolded protein, purified from misfolded forms, was eluted directly from the column at a concentration of 3 mg/ml. This technique integrates the dithiothreitol removal, refolding, concentration and purification steps, achieving a high level of process simplification and automation, and a significant saving in reagent costs when scaled. Importantly, the current result suggests that it is possible to controllably refold disulfide-bonded proteins using common and inexpensive matrices, and that it is not always necessary to control protein-surface interactions using affinity tags and expensive chromatographic matrices. Moreover, it is possible to strictly control the oxidative refolding environment once denatured protein is bound to the ion-exchange column, thus allowing precisely controlled oxido-shuffling. (c) 2005 Elsevier B.V. All rights reserved.

NMR microscopy: A tool for measuring monomer diffusion in supercritical CO2

The diffusion of styrene into linear low density polyethylene in a solution of supercritical CO2 was investigated using NMR microimaging. For both pure styrene and styrene dissolved in supercritical CO2, the diffusion was found to follow Fickian kinetics. Supercritical CO2 was found to enhance the rate and extent of diffusion of styrene into the substrate by up to three times under the conditions of this investigation, compared to pure styrene. NMR imaging was used to measure the concentration profiles of the styrene penetrants in real time, and the results were fitted to a Fickian model for diffusion. At a CO2 pressure of 150 bar and temperature of 40 degrees C, the diffusion coefficient of a 30 wt-% solution of styrene into LLDPE was calculated to be 1 X 10(-11) m(2). s(-1). This is significantly faster than the diffusion coefficient measured for pure styrene diffusion at 40 degrees C (3 x 10(-12) m(2). s(-1)). The diffusion coefficients determined by gravimetric analysis were slightly higher than those determined by the imaging method. This was probably due to residual styrene and/or polystyrene adhering to the surface of the films in the gravimetric technique.

Web based expert elicitation of uncertainties in environmental model inputs

When constructing and using environmental models, it is typical that many of the inputs to the models will not be known perfectly. In some cases, it will be possible to make observations, or occasionally physics-based uncertainty propagation, to ascertain the uncertainty on these inputs. However, such observations are often either not available or even possible, and another approach to characterising the uncertainty on the inputs must be sought. Even when observations are available, if the analysis is being carried out within a Bayesian framework then prior distributions will have to be specified. One option for gathering or at least estimating this information is to employ expert elicitation. Expert elicitation is well studied within statistics and psychology and involves the assessment of the beliefs of a group of experts about an uncertain quantity, (for example an input / parameter within a model), typically in terms of obtaining a probability distribution. One of the challenges in expert elicitation is to minimise the biases that might enter into the judgements made by the individual experts, and then to come to a consensus decision within the group of experts. Effort is made in the elicitation exercise to prevent biases clouding the judgements through well-devised questioning schemes. It is also important that, when reaching a consensus, the experts are exposed to the knowledge of the others in the group. Within the FP7 UncertWeb project (http://www.uncertweb.org/), there is a requirement to build a Webbased tool for expert elicitation. In this paper, we discuss some of the issues of building a Web-based elicitation system - both the technological aspects and the statistical and scientific issues. In particular, we demonstrate two tools: a Web-based system for the elicitation of continuous random variables and a system designed to elicit uncertainty about categorical random variables in the setting of landcover classification uncertainty. The first of these examples is a generic tool developed to elicit uncertainty about univariate continuous random variables. It is designed to be used within an application context and extends the existing SHELF method, adding a web interface and access to metadata. The tool is developed so that it can be readily integrated with environmental models exposed as web services. The second example was developed for the TREES-3 initiative which monitors tropical landcover change through ground-truthing at confluence points. It allows experts to validate the accuracy of automated landcover classifications using site-specific imagery and local knowledge. Experts may provide uncertainty information at various levels: from a general rating of their confidence in a site validation to a numerical ranking of the possible landcover types within a segment. A key challenge in the web based setting is the design of the user interface and the method of interacting between the problem owner and the problem experts. We show the workflow of the elicitation tool, and show how we can represent the final elicited distributions and confusion matrices using UncertML, ready for integration into uncertainty enabled workflows.We also show how the metadata associated with the elicitation exercise is captured and can be referenced from the elicited result, providing crucial lineage information and thus traceability in the decision making process.

Product cipher negotiation with on-line evaluation for private communication over computer networks

A method is proposed to offer privacy in computer communications, using symmetric product block ciphers. The security protocol involved a cipher negotiation stage, in which two communicating parties select privately a cipher from a public cipher space. The cipher negotiation process includes an on-line cipher evaluation stage, in which the cryptographic strength of the proposed cipher is estimated. The cryptographic strength of the ciphers is measured by confusion and diffusion. A method is proposed to describe quantitatively these two properties. For the calculation of confusion and diffusion a number of parameters are defined, such as the confusion and diffusion matrices and the marginal diffusion. These parameters involve computationally intensive calculations that are performed off-line, before any communication takes place. Once they are calculated, they are used to obtain estimation equations, which are used for on-line, fast evaluation of the confusion and diffusion of the negotiated cipher. A technique proposed in this thesis describes how to calculate the parameters and how to use the results for fast estimation of confusion and diffusion for any cipher instance within the defined cipher space.

The effect of thermochemical treatment on the fatigue properties of alloy steels

The effect of thermochemical treatment namely carburising on the fatigue behaviour of one carbon and two alloy steels has been studied in rotating and unidirectional bending. The effect of carbon profile on the unidirect¬ional bending fatigue strength of 63SA14 was assessed, and it was found that single stage carburising with a surface carbon content of 0.8% has resulted in a higher fatigue strength than other types of carbon profiles. Residual stresses and other metallurgical variables arising from different carbon profiles, were also considered. The highest compressive stresses h~e resulted from boost-diffuse-carburising. On the other hand surface decarburisation was associated with tensile residual stresses and a reduced fatigue strength. Retained austenite was found to be detrimental in unidirectional bending fatigue; however its presence in carburised 83SAIS did not seem to influence the rotating bending fatigue strength. Carbide particles in globular and/or intergranular form were detrimental to compressive residual stresses; the unidirectional bending fatigue strength is markedly lowered. The highest fatigue strength was accomplished by vacuum carburising. The absence of internal oxidation was the key factor in the increased fatigue strength; the presence of uniformly distributed fine carbide particles did not upset the superior fatigue strength of vacuum carburised pieces. The effect of mean stress on the fatigue strength of carburised 63SA14 was studied. Increasing the mean stress as would be expected resulted in a decreased fatigue strength. Carburisation showed its advantages at low mean stress, but at high mean stress it offers little advantage over the uncarburised hardened conditions. Notch effect was also studied in unidirectional bending of carburised 080MlS. The general trend showed that the fatigue strength decreases with increasing the stress concentration factor. But different carburising conditions have different effect on notch sensitivity.

FemtoNewton force sensing with optically trapped nanotubes

We extract the distribution of both center-of-mass and angular fluctuations from three-dimensional tracking of optically trapped nanotubes. We measure the optical force and torque constants from autocorrelation and cross-correlation of the tracking signals. This allows us to isolate the angular Brownian motion. We demonstrate that nanotubes enable nanometer spatial and femtonewton force resolution in photonic force microscopy, the smallest to date. This has wide implications in nanotechnology, biotechnology, nanofluidics, and material science.

A perturbative analysis of dispersion-managed solitons

We study soliton solutions of the path-averaged propagation equation governing the transmission of dispersion-managed (DM) optical pulses in the (practical) limit when residual dispersion and nonlinearity only slightly affect the pulse dynamics over one compensation period. In the case of small dispersion map strengths, the averaged pulse dynamics is governed by a perturbed form of the nonlinear Schrödinger equation; applying a perturbation theory – elsewhere developed – based on inverse scattering theory, we derive an analytic expression for the envelope of the DM soliton. This expression correctly predicts the power enhancement arising from the dispersion management. Theoretical results are verified by direct numerical simulations.

A perturbative analysis of dispersion-managed solitons

We study soliton solutions of the path-averaged propagation equation governing the transmission of dispersion-managed (DM) optical pulses in the (practical) limit when residual dispersion and nonlinearity only slightly affect the pulse dynamics over one compensation period. In the case of small dispersion map strengths, the averaged pulse dynamics is governed by a perturbed form of the nonlinear Schrödinger equation; applying a perturbation theory – elsewhere developed – based on inverse scattering theory, we derive an analytic expression for the envelope of the DM soliton. This expression correctly predicts the power enhancement arising from the dispersion management. Theoretical results are verified by direct numerical simulations.

Thermal stability of sputter-deposited 330 austenitic stainless-steel thin films with nanoscale growth twins

We have explored the thermal stability of nanoscale growth twins in sputter-deposited 330 stainless-steel (SS) films by vacuum annealing up to 500 °C. In spite of an average twin spacing of only 4 nm in the as-deposited films, no detectable variation in the twin spacing or orientation of twin interfaces was observed after annealing. An increase in the average columnar grain size was observed after annealing. The hardness of 330 SS films increases after annealing, from 7 GPa for as-deposited films to around 8 GPa for annealed films, while the electrical resistivity decreases slightly after annealing. The changes in mechanical and electrical properties after annealing are interpreted in terms of the corresponding changes in the residual stress and microstructure of the films. © 2005 American Institute of Physics.

In situ studies of structure-reactivity relations in biodiesel synthesis over nanocrystalline MgO

High temperature processing of solvothermally synthesised MgO nanoparticles promotes striking changes in their morphology, and surface chemical and electronic structure. As-prepared NanoMgO comprised ∼4 nm cubic periclase nanocrystals, interspersed within an amorphous Mg(OH)(OCH3) matrix. These crystallites appear predominantly (1 0 0) terminated, and the overall material exhibits carbonate and hydroxyl surface functionalities of predominantly weak/moderate base character. Heating promotes gradual crystallisation and growth of the MgO nanoparticles, and concomitant loss of Mg(OH)(OCH3). In situ DRIFTS confirms the residual precursor and surface carbonate begin to decompose above 300 °C, while in situ XPS shows these morphological changes are accompanied by the disappearance of surface hydroxyl/methoxide species and genesis of O- centres which enhance both the surface density and basicity of the resulting stepped and defective MgO nanocrystals. The catalytic performance in tributyrin transesterification with methanol is directly proportional to the density of strong surface base sites. © 2010 Elsevier B.V. All rights reserved.

Some Examples of Rigid Representations

*Research partially supported by INTAS grant 97-1644.

A decision support framework for infrastructure maintenance investment decision-making

Infrastructure management agencies are facing multiple challenges, including aging infrastructure, reduction in capacity of existing infrastructure, and availability of limited funds. Therefore, decision makers are required to think innovatively and develop inventive ways of using available funds. Maintenance investment decisions are generally made based on physical condition only. It is important to understand that spending money on public infrastructure is synonymous with spending money on people themselves. This also requires consideration of decision parameters, in addition to physical condition, such as strategic importance, socioeconomic contribution and infrastructure utilization. Consideration of multiple decision parameters for infrastructure maintenance investments can be beneficial in case of limited funding. Given this motivation, this dissertation presents a prototype decision support framework to evaluate trade-off, among competing infrastructures, that are candidates for infrastructure maintenance, repair and rehabilitation investments. Decision parameters' performances measured through various factors are combined to determine the integrated state of an infrastructure using Multi-Attribute Utility Theory (MAUT). The integrated state, cost and benefit estimates of probable maintenance actions are utilized alongside expert opinion to develop transition probability and reward matrices for each probable maintenance action for a particular candidate infrastructure. These matrices are then used as an input to the Markov Decision Process (MDP) for the finite-stage dynamic programming model to perform project (candidate)-level analysis to determine optimized maintenance strategies based on reward maximization. The outcomes of project (candidate)-level analysis are then utilized to perform network-level analysis taking the portfolio management approach to determine a suitable portfolio under budgetary constraints. The major decision support outcomes of the prototype framework include performance trend curves, decision logic maps, and a network-level maintenance investment plan for the upcoming years. The framework has been implemented with a set of bridges considered as a network with the assistance of the Pima County DOT, AZ. It is expected that the concept of this prototype framework can help infrastructure management agencies better manage their available funds for maintenance.

A política nacional de educação ambiental: limites e desafios para a sua efetivação na Universidade Federal do Rio Grande do Norte

The Environmental Education is a plural and diversified knowledge field, composed by a group of social agents of specific State, government and civil society sectors, with different world views and theoretical matrices that exercised and influenced its genealogy and constitutive dynamic. The Environmental Education – while specific knowledge field – has been produced, systematized and diffused in Brazil in the last decades by official State organisms, through public policies, as well as by social movements of popular education, and constitutes a large and historical movement about the environmental question, that was worldwide projected in the 1960 decade (the environmentalist movement). Concerning the creation of public policies and specific programs to the Environmental Education, in the scope of the governmental initiative, the approval of the National Policy of Environmental Education, by means of the Law n. 9.795/99 – together with its regulatory decree, the Decree nº 4.281 – represents the consolidation of a inclusion process of the environmental dimension in the educational field. These normative acts, beyond charging the public power with the incumbency to define public policies that incorporate the environmental dimension and to promote the Environmental Education in all education levels, also charge the educational institution with the duty of promoting this component in an integrated and articulated way with the educational programs that such institutions develop. In this context, it is aimed to identify and analyze under the light of the dialectical and historical materialism, the practices and concepts developed under the Federal University of Rio Grande do Norte (UFRN), with regard to its institutional policy on Environmental Education, relating it to establishing the Pnea. The analysis by means specific legislation on the subject and institutional documents UFRN. The research showed that, in general, the National Environmental Education Policy has limits to its effectiveness given the absence of specific funding for this purpose and the little government involvement in that the Brazilian State assumes, by through effective policies, the material conditions of financing actions with respect to this field of education. The fragmented and disjointed way the analyzed actions are developed constitutes a limit to the challenge for the UFRN implement, consistently and objectively, an Environmental Education policy, which can be monitored and evaluated as an effective public policy, both landmarks major goal of own Pnea as the demands of development whose agenda a critical environmental perspective.

Modelo para reconstrução 3D de cenas baseado em imagens

3D Reconstruction is the process used to obtain a detailed graphical model in three dimensions that represents some real objectified scene. This process uses sequences of images taken from the scene, so it can automatically extract the information about the depth of feature points. These points are then highlighted using some computational technique on the images that compose the used dataset. Using SURF feature points this work propose a model for obtaining depth information of feature points detected by the system. At the ending, the proposed system extract three important information from the images dataset: the 3D position for feature points; relative rotation and translation matrices between images; the realtion between the baseline for adjacent images and the 3D point accuracy error found.

Um estudo sobre polinômios matriciais

This research work aims to make a study of the algebraic theory of matrix monic polynomials, as well as the definitions, concepts and properties with respect to block eigenvalues, block eigenvectors and solvents of P(X). We investigte the main relations between the matrix polynomial and the Companion and Vandermonde matrices. We study the construction of matrix polynomials with certain solvents and the extention of the Power Method, to calculate block eigenvalues and solvents of P(X). Through the relationship between the dominant block eigenvalue of the Companion matrix and the dominant solvent of P(X) it is possible to obtain the convergence of the algorithm for the dominant solvent of the matrix polynomial. We illustrate with numerical examples for diferent cases of convergence.