João de Barros e o Erasmismo

pp. 19-26

Detection of cardiac biomarker proteins using a disposable based on a molecularly imprinted polymer grafted onto graphite

A low-cost disposable was developed for rapid detection of the protein biomarker myoglobin (Myo) as a model analyte. A screen printed electrode was modified with a molecularly imprinted material grafted on a graphite support and incorporated in a matrix composed of poly(vinyl chloride) and the plasticizer o-nitrophenyloctyl ether. The protein-imprinted material (PIM) was produced by growing a reticulated polymer around a protein template. This is followed by radical polymerization of 4-styrenesulfonic acid, 2-aminoethyl methacrylate hydrochloride, and ethylene glycol dimethacrylate. The polymeric layer was then covalently bound to the graphitic support, and Myo was added during the imprinting stage to act as a template. Non-imprinted control materials (CM) were also prepared by omitting the Myo template. Morphological and structural analysis of PIM and CM by FTIR, Raman, and SEM/EDC microscopies confirmed the modification of the graphite support. The analytical performance of the SPE was assessed by square wave voltammetry. The average limit of detection is 0.79 μg of Myo per mL, and the slope is −0.193 ± 0.006 μA per decade. The SPE-CM cannot detect such low levels of Myo but gives a linear response at above 7.2 μg · mL−1, with a slope of −0.719 ± 0.02 μA per decade. Interference studies with hemoglobin, bovine serum albumin, creatinine, and sodium chloride demonstrated good selectivity for Myo. The method was successfully applied to the determination of Myo urine and is conceived to be a promising tool for screening Myo in point-of-care patients with ischemia.

Protein-responsive polymers for point-of-care detection of cardiac biomarker

This work describes a novel use for the polymeric film, poly(o-aminophenol) (PAP) that was made responsive to a specific protein. This was achieved through templated electropolymerization of aminophenol (AP) in the presence of protein. The procedure involved adsorbing protein on the electrode surface and thereafter electroploymerizing the aminophenol. Proteins embedded at the outer surface of the polymeric film were digested by proteinase K and then washed away thereby creating vacant sites. The capacity of the template film to specifically rebind protein was tested with myoglobin (Myo), a cardiac biomarker for ischemia. The films acted as biomimetic artificial antibodies and were produced on a gold (Au) screen printed electrode (SPE), as a step towards disposable sensors to enable point-of-care applications. Raman spectroscopy was used to follow the surface modification of the Au-SPE. The ability of the material to rebind Myo was measured by electrochemical techniques, namely electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV). The devices displayed linear responses to Myo in EIS and SWV assays down to 4.0 and 3.5 μg/mL, respectively, with detection limits of 1.5 and 0.8 μg/mL. Good selectivity was observed in the presence of troponin T (TnT) and creatine kinase (CKMB) in SWV assays, and accurate results were obtained in applications to spiked serum. The sensor described in this work is a potential tool for screening Myo in point-of-care due to the simplicity of fabrication, disposability, short time response, low cost, good sensitivity and selectivity.

Electrochemical biosensor based on biomimetic material for myoglobin detection

A novel reusable molecularly imprinted polymer (MIP) assembled on a polymeric layer of carboxylated poly(vinyl chloride) (PVCsingle bondCOOH) for myoglobin (Myo) detection was developed. This polymer was casted on the gold working area of a screen printed electrode (Au-SPE), creating a novel disposable device relying on plastic antibodies. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and Fourier transform infrared spectroscopy (FTIR) studies confirmed the surface modification. The MIP/Au-SPE devices displayed a linear behaviour in EIS from 0.852 to 4.26 μg mL−1, of positive slope 6.50 ± 1.48 (kΩ mL μg−1). The limit of detection was 2.25 μg mL−1. Square wave voltammetric (SWV) assays were made in parallel and showed linear responses between 1.1 and 2.98 μg mL−1. A current decrease was observed against Myo concentration, producing average slopes of −0.28 ± 0.038 μA mL μg−1. MIP/Au-SPE also showed good results in terms of selectivity. The error% found for each interfering species were 7% for troponin T (TnT), 11% for bovine serum albumin (BSA) and 2% for creatine kinase MB (CKMB), respectively. Overall, the technical modification over the Au-SPE was found a suitable approach for screening Myo in biological fluids.

Figuras do Dom (em torno do miserere de João Morais Barbosa

pp. 223-228

Novel biosensing device for point-of-care applications with plastic antibodies grown on Au-Screen Printed Electrodes

A gold screen printed electrode (Au-SPE) was modified by merging Molecular Imprinting and Self-Assembly Monolayer techniques for fast screening cardiac biomarkers in point-of-care (POC). For this purpose, Myoglobin (Myo) was selected as target analyte and its plastic antibody imprinted over a glutaraldehyde (Glu)/cysteamine (Cys) layer on the gold-surface. The imprinting effect was produced by growing a reticulated polymer of acrylamide (AAM) and N,N′-methylenebisacrylamide (NNMBA) around the Myo template, covalently attached to the biosensing surface. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies were carried out in all chemical modification steps to confirm the surface changes in the Au-SPE. The analytical features of the resulting biosensor were studied by different electrochemical techniques, including EIS, square wave voltammetry (SWV) and potentiometry. The limits of detection ranged from 0.13 to 8 μg/mL. Only potentiometry assays showed limits of detection including the cut-off Myo levels. Quantitative information was also produced for Myo concentrations ≥0.2 μg/mL. The linear response of the biosensing device showed an anionic slope of ~70 mV per decade molar concentration up to 0.3 μg/mL. The interference of coexisting species was tested and good selectivity was observed. The biosensor was successfully applied to biological fluids.

Smart Plastic Antibody Material (SPAM) tailored on disposable screen printed electrodes for protein recognition: application to Myoglobin detection

This work introduces two major changes to the conventional protocol for designing plastic antibodies: (i) the imprinted sites were created with charged monomers while the surrounding environment was tailored using neutral material; and (ii) the protein was removed from its imprinted site by means of a protease, aiming at preserving the polymeric network of the plastic antibody. To our knowledge, these approaches were never presented before and the resulting material was named here as smart plastic antibody material (SPAM). As proof of concept, SPAM was tailored on top of disposable gold-screen printed electrodes (Au-SPE), following a bottom-up approach, for targeting myoglobin (Myo) in a point-of-care context. The existence of imprinted sites was checked by comparing a SPAM modified surface to a negative control, consisting of similar material where the template was omitted from the procedure and called non-imprinted materials (NIMs). All stages of the creation of the SPAM and NIM on the Au layer were followed by both electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). AFM imaging was also performed to characterize the topography of the surface. There are two major reasons supporting the fact that plastic antibodies were effectively designed by the above approach: (i) they were visualized for the first time by AFM, being present only in the SPAM network; and (ii) only the SPAM material was able to rebind to the target protein and produce a linear electrical response against EIS and square wave voltammetry (SWV) assays, with NIMs showing a similar-to-random behavior. The SPAM/Au-SPE devices displayed linear responses to Myo in EIS and SWV assays down to 3.5 μg/mL and 0.58 μg/mL, respectively, with detection limits of 1.5 and 0.28 μg/mL. SPAM materials also showed negligible interference from troponin T (TnT), bovine serum albumin (BSA) and urea under SWV assays, showing promising results for point-of-care applications when applied to spiked biological fluids.

Surface Imprinting Approach on Screen Printed Electrodes Coated with Carboxylated PVC for Myoglobin detection with Electrochemical Transduction

A novel surface molecularly-imprinted (MI) material to detect myoglobin (Myo) using gold screen printed electrodes (SPE) was developed. The sensitive detection was carry out by introducing a carboxylic polyvinyl chloride (PVC-COOH) layer on gold SPE surface. Myo was attached to the surface of gold SPE/PVC-COOH and the vacant spaces around it were filled by polymerizing acrylamide and N,N-methylenebisacrylamide (cross-linker). This polymerization was initiated by ammonium persulphate. After removing the template, the obtained material was able to rebind Myo and discriminate it among other interfering species. Various characterization techniques including electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) confirmed the surface modification. This sensor seemed a promising tool for screening Myo in point-of-care.

Artificial antibodies for troponin T by its imprinting on the surface of multiwalled carbon nanotubes: Its use as sensory surfaces

A novel artificial antibody for troponin T (TnT) was synthesized by molecular imprint (MI) on the surface of multiwalled carbon nanotubes (MWCNT). This was done by attaching TnT to the MWCNT surface, and filling the vacant spaces by polymerizing under mild conditions acrylamide (monomer) in N,N′-methylenebisacrylamide (cross-linker) and ammonium persulphate (initiator). After removing the template, the obtained biomaterial was able to rebind TnT and discriminate it among other interfering species. Stereochemical recognition of TnT was confirmed by the non-rebinding ability displayed by non-imprinted (NI) materials, obtained by imprinting without a template. SEM and FTIR analysis confirmed the surface modification of the MWCNT. The ability of this biomaterial to rebind TnT was confirmed by including it as electroactive compound in a PVC/plasticizer mixture coating a wire of silver, gold or titanium. Anionic slopes of 50 mV decade−1 were obtained for the gold wire coated with MI-based membranes dipped in HEPES buffer of pH 7. The limit of detection was 0.16 μg mL−1. Neither the NI-MWCNT nor the MWCNT showed the ability to recognize the template. Good selectivity was observed against creatinine, sucrose, fructose, myoglobin, sodium glutamate, thiamine and urea. The sensor was tested successfully on serum samples. It is expected that this work opens new horizons on the design of new artificial antibodies for complex protein structures.

Myoglobin-biomimetic electroactive materials made by surface molecular imprinting on silica beads and their use as ionophores in polymeric membranes for potentiometric transduction

Myoglobin (Mb) is among the cardiac biomarkers playing a major role in urgent diagnosis of cardiovascular diseases. Its monitoring in point-of-care is therefore fundamental. Pursuing this goal, a novel biomimetic ionophore for the potentiometric transduction of Mb is presented. It was synthesized by surface molecular imprinting (SMI) with the purpose of developing highly efficient sensor layers for near-stereochemical recognition of Mb. The template (Mb) was imprinted on a silane surface that was covalently attached to silica beads by means of self-assembled monolayers. First the silica was modified with an external layer of aldehyde groups. Then, Mb was attached by reaction with its amine groups (on the external surface) and subsequent formation of imine bonds. The vacant places surrounding Mb were filled by polymerization of the silane monomers 3-aminopropyltrimethoxysilane (APTMS) and propyltrimethoxysilane (PTMS). Finally, the template was removed by imine cleavage after treatment with oxalic acid. The results materials were finely dispersed in plasticized PVC selective membranes and used as ionophores in potentiometric transduction. The best analytical features were found in HEPES buffer of pH 4. Under this condition, the limits of detection were of 1.3 × 10−6 mol/L for a linear response after 8.0 × 10−7 mol/L with an anionic slope of −65.9 mV/decade. The imprinting effect was tested by preparing non-imprinted (NI) particles and employing these materials as ionophores. The resulting membranes showed no ability to detect Mb. Good selectivity was observed towards creatinine, sacarose, fructose, galactose, sodium glutamate, and alanine. The analytical application was conducted successfully and showed accurate and precise results.

Proposta de tratamento documental do arquivo da brigada de estudos de azulejaria da Biblioteca de Arte da Fundação Calouste Gulbenkian

Relatório de Estágio apresentado para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Ciências da Informação e da Documentação - Área de Especialização em Biblioteconomia

Resident's perceptions on impacts of hosting the “Guimarães 2012 European Capital of Culture

. Residents tend to have high expectations about the benefits of hosting a mega‐event. So, it was not surprising that the nomination of Guimarães, Portugal, as the 2012 European Capital of Culture (2012 ECOC) had raised great expectations in the local community towards its socio‐economic and cultural benefits. The present research was designed to examine the Guimarães residents’ perceptions on the impacts of hosting the 2012 ECOC approached in two different time schedules, the pre‐ and the post‐event, trying to capture the evolution of the residents` evaluation of its impacts. For getting the data, two surveys were applied to Guimarães` residents, one in the pre‐event phase, in 2011, and another in the post‐event phase, in 2013. This approach is uncommonly applied to Portugal data and it is even the first time it was done to a Portuguese European Capital of Culture. After a factor analysis, the results of t‐tests indicate that there were significant differences (p<0.05) between the samples from the pre‐ and post‐2012 ECOC on two positive impact factors (Community’ benefits and Residents’ benefits) and one negative impact factor (Economic, social and environmental costs). Respondents also showed a negative perception of the impacts in all dimensions, except Changes in habits of Guimarães residents. 

Ex-ante evaluation by the media of the Guimarães 2012 European Capital of Culture

The nomination of Guimarães to host the 2012 European Capital of Culture (ECC) has put on the agenda of the city the need of measuring the effects that the implementation of this mega event could have in it and in the municipality a whole. The balance of the benefits and costs and an extended community involvement tend to reduce negative impacts and enhance positive ones. This chapter analyzes the involvement of population and local associations in the planning and organization of the 2012 Guimarães European Capital of Culture, using the coverage made during 2011 by local and national press of the mega event. A content analysis of the news published covering the period between January and December 2011 and using three newspapers was conducted. From those, two were local and weekly newspapers and one was a national daily one. Looking to data results, it can be concluded that it was poor the community involvement and, also, the one of the cultural associations in the organizations of the 2012 ECC. A strong negative reaction to the model choose to plan the mega event conducted by official organizers was found, which has cast doubts on the desirable participation of the residents and, consequently, on the success of the mega event, especially in a perspective of a medium and long term effects.

Potenciais impactos para Guimarães do acolhimento da Capital Europeia da Cultura 2012

A nomeação de Guimarães como Capital Europeia da Cultura (CEC), em 2012, suscitou grandes expectativas na comunidade local, nos territórios limítrofes e, de um modo geral, na população portuguesa. No entanto, muitos dos potenciais impactos são ainda pouco perceptíveis pela maior parte das pessoas, se bem que muitas delas antecipem que os impactos sejam positivos. A presente comunicação pretende aferir, de forma prospectiva, os possíveis impactos do acolhimento por Guimarães da CEC, em 2012. Por referência a experiências anteriores de organização de outras CEC, procura-se identificar alguns dos efeitos, em termos de atracção de turistas (nacionais e internacionais), das actividades culturais programadas, disponibilidades orçamentais e audiências, bem como aferir do potencial impacto económico e do legado do evento para o futuro da cidade. Com base na análise efectuada, pode concluir-se que Guimarães CEC 2012 oferece um potencial de atracção para o Norte do país e região da Galiza, possui um orçamento, por habitante, comparável a outras CEC e beneficia do empenhamento dos agentes locais. Menos positiva é a distância da cidade a dois dos principais destinos turísticos nacionais (Lisboa e Algarve), a extensão da programação cultural do evento, o fraco envolvimento dos agentes culturais locais no programa de actividades do evento e a inexistência de espaços museológicos de referência internacional. Em contrapartida, destaca-se a existência de ligações aéreas de baixo custo a várias cidades Europeias através do aeroporto do Porto, a possibilidade de reposicionar a imagem da cidade ao nível nacional e internacional, a disponibilização de fundos para a revitalização urbana do centro histórico e o aumento esperado do número de turistas (nacionais e estrangeiros). A recessão económica (nacional e Europeia), as dificuldades orçamentais, a pouca ambição da política de angariação de patrocínios e de recursos financeiros privados e a sustentabilidade do evento no futuro são algumas ameaças que poderão comprometer o sucesso de Guimarães CEC 2012

Análise de sinais de voz para Detecção de patologias da Laringe Utilizando Entropia de Tsallis

Este trabalho apresenta uma nova metodologia para se comparar com trabalhos anteriores realizados por Scalassara (2009) e santos (2011), para o diagnóstico de patologias da laringe por meio da análise da voz. Nos trabalhos foram utilizados as Entropias shannon, Rényi e suas Entropias relativas para se obter uma classificação a partir do sinal de voz. desta maneira detecta-se a pessoa está doente e distingue-se entre duas patologias, nódulos nas pregas vocais ou o edema Reinke. Neste estudo é utilizada a entropia de Tsaillis, com o objetivo de se avaliar um melhor método de avaliação para o mesmo problema. o aperfeiçoamento desta técnica de diagnóstico é uma interessante alternativa às práticas atuais, sendo a principal diferença não consistir num exame invasivo e desta forma desconfortável para o paciente. Os resultados obtidos foram satisfatórios e foi possível determinar com precisão sinais saudáveis de sinais com as duas patologias contempladas pelo estudo.