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.

Specific label-free and real-time detection of oxidized low density lipoprotein (oxLDL) using an immunosensor with three monoclonal antibodies

Increased levels of plasma oxLDL, which is the oxidized fraction of Low Density Lipoprotein (LDL), are associated with atherosclerosis, an inflammatory disease, and the subsequent development of severe cardiovascular diseases that are today a major cause of death in modern countries. It is therefore important to find a reliable and fast assay to determine oxLDL in serum. A new immunosensor employing three monoclonal antibodies (mAbs) against oxLDL is proposed in this work as a quick and effective way to monitor oxLDL. The oxLDL was first employed to produce anti-oxLDL monoclonal antibodies by hybridoma cells that were previously obtained. The immunosensor was set-up by selfassembling cysteamine (Cyst) on a gold (Au) layer (4 mm diameter) of a disposable screen-printed electrode. Three mAbs were allowed to react with N-hydroxysuccinimide (NHS) and ethyl(dimethylaminopropyl)carbodiimide (EDAC), and subsequently incubated in the Au/Cys. Albumin from bovine serum (BSA) was immobilized further to ensure that other molecules apart from oxLDL could not bind to the electrode surface. All steps were followed by various characterization techniques such as electrochemical impedance spectroscopy (EIS) and square wave voltammetry (SWV). The analytical operation of the immunosensor was obtained by incubating the sensing layer of the device in oxLDL for 15 minutes, prior to EIS and SWV. This was done by using standard oxLDL solutions prepared in foetal calf serum, in order to simulate patient's plasma with circulating oxLDL. A sensitive response was observed from 0.5 to 18.0 mg mL 1 . The device was successfully applied to determine the oxLDL fraction in real serum, without prior dilution or necessary chemical treatment. The use of multiple monoclonal antibodies on a biosensing platform seemed to be a successful approach to produce a specific response towards a complex multi-analyte target, correlating well with the level of oxLDL within atherosclerosis disease, in a simple, fast and cheap way.

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.

Protein–polyphenol interaction on silica beads for astringency tests based on eye, photography or reflectance detection modes

Astringency is an organoleptic property of beverages and food products resulting mainly from the interaction of salivary proteins with dietary polyphenols. It is of great importance to consumers, but the only effective way of measuring it involves trained sensorial panellists, providing subjective and expensive responses. Concurrent chemical evaluations try to screen food astringency, by means of polyphenol and protein precipitation procedures, but these are far from the real human astringency sensation where not all polyphenol–protein interactions lead to the occurrence of precipitate. Here, a novel chemical approach that tries to mimic protein–polyphenol interactions in the mouth is presented to evaluate astringency. A protein, acting as a salivary protein, is attached to a solid support to which the polyphenol binds (just as happens when drinking wine), with subsequent colour alteration that is fully independent from the occurrence of precipitate. Employing this simple concept, Bovine Serum Albumin (BSA) was selected as the model salivary protein and used to cover the surface of silica beads. Tannic Acid (TA), employed as the model polyphenol, was allowed to interact with the BSA on the silica support and its adsorption to the protein was detected by reaction with Fe(III) and subsequent colour development. Quantitative data of TA in the samples were extracted by colorimetric or reflectance studies over the solid materials. The analysis was done by taking a regular picture with a digital camera, opening the image file in common software and extracting the colour coordinates from HSL (Hue, Saturation, Lightness) and RGB (Red, Green, Blue) colour model systems; linear ranges were observed from 10.6 to 106.0 μmol L−1. The latter was based on the Kubelka–Munk response, showing a linear gain with concentrations from 0.3 to 10.5 μmol L−1. In either of these two approaches, semi-quantitative estimation of TA was enabled by direct eye comparison. The correlation between the levels of adsorbed TA and the astringency of beverages was tested by using the assay to check the astringency of wines and comparing these to the response of sensorial panellists. Results of the two methods correlated well. The proposed sensor has significant potential as a robust tool for the quantitative/semi-quantitative evaluation of astringency in wine.

Novel LTCC-potentiometric microfluidic device for biparametric analysis of organic compounds carrying plastic antibodies as ionophores: Application to sulfamethoxazole and trimethoprim

Monitoring organic environmental contaminants is of crucial importance to ensure public health. This requires simple, portable and robust devices to carry out on-site analysis. For this purpose, a low-temperature co-fired ceramics (LTCC) microfluidic potentiometric device (LTCC/μPOT) was developed for the first time for an organic compound: sulfamethoxazole (SMX). Sensory materials relied on newly designed plastic antibodies. Sol–gel, self-assembling monolayer and molecular-imprinting techniques were merged for this purpose. Silica beads were amine-modified and linked to SMX via glutaraldehyde modification. Condensation polymerization was conducted around SMX to fill the vacant spaces. SMX was removed after, leaving behind imprinted sites of complementary shape. The obtained particles were used as ionophores in plasticized PVC membranes. The most suitable membrane composition was selected in steady-state assays. Its suitability to flow analysis was verified in flow-injection studies with regular tubular electrodes. The LTCC/μPOT device integrated a bidimensional mixer, an embedded reference electrode based on Ag/AgCl and an Ag-based contact screen-printed under a micromachined cavity of 600 μm depth. The sensing membranes were deposited over this contact and acted as indicating electrodes. Under optimum conditions, the SMX sensor displayed slopes of about −58.7 mV/decade in a range from 12.7 to 250 μg/mL, providing a detection limit of 3.85 μg/mL and a sampling throughput of 36 samples/h with a reagent consumption of 3.3 mL per sample. The system was adjusted later to multiple analyte detection by including a second potentiometric cell on the LTCC/μPOT device. No additional reference electrode was required. This concept was applied to Trimethoprim (TMP), always administered concomitantly with sulphonamide drugs, and tested in fish-farming waters. The biparametric microanalyzer displayed Nernstian behaviour, with average slopes −54.7 (SMX) and +57.8 (TMP) mV/decade. To demonstrate the microanalyzer capabilities for real applications, it was successfully applied to single and simultaneous determination of SMX and TMP in aquaculture waters.

Protecção radiológica do doente em tomografia computorizada

RESUMO - A preocupação com os conhecimentos, atitudes, crenças e práticas, no que concerne à utilização de radiações ionizantes para fins de diagnóstico, e a sensibilização de todos os agentes envolvidos, médicos, técnicos, físicos, utentes e responsáveis pela Saúde Publica, relativamente aos níveis de radiação emitida nos exames de Tomografia Computorizada (TC), assume particular importância no domínio da Saúde Pública, na medida em que é necessário influenciar o desenvolvimento de práticas que promovam, auditem e garantam a prestação do controlo da qualidade radiológica e dosimétrica nos serviços de Radiologia a nível Nacional. Para tal, e no âmbito da publicação de estudos já realizados ao nível da União Europeia, ―Orientações Europeias dos Critérios de Qualidade para a Tomografia Computorizada (1999) ‖, é proposto estabelecer orientações na realização de estudos que permitam, numa primeira fase, estabelecer a comparação com os resultados obtidos pelos mecanismos de Controlo da Qualidade (CQ), analisar e proceder aos ajustes (se necessário) e, numa segunda fase, implementar uma moldura sistemática de avaliação periódica dos níveis de dose de radiação por exame TC e que permita a monitorização dos dados. Nesse sentido, propõe-se a realização de um Estudo Nacional que envolva a rede hospitalar pública, privada e universitária, partindo da metodologia utilizada em estudos prévios noutros países da Europa, como seja, selecção do equipamento de TC existente na Instituição Hospitalar, onde serão reunidas informações através do preenchimento de questionários relativos ao equipamento a utilizar. Serão recolhidos dados relativos ao utente, ao equipamento e parâmetros de aquisição de imagem, que permitam identificar os níveis de referência de diagnóstico (NRD) em TC, na realidade Portuguesa. Foi efectuado um estudo piloto numa instituição EPE e os valores obtidos não são significativos, nem podem assumir valor preditivo dado o reduzido tamanho da amostra. Apesar disso, sugerem a existência de parâmetros que podem ser alterados e com isso podem fazer variar a dose de radiação utilizada. ENSP/UNL Maria de Fátima Vaz de Carvalho 5 Espera-se obter com este estudo, como foi referido, a base do estabelecimento dos NRD em TC em Portugal. ----------------- ABSTRACT - The purpose of this study, in an empirical point of view, emerges from concern with the knowledge, attitudes, beliefs and practices regarding the use of ionizing radiation for diagnostic purposes and awareness of all actors involved, medical physical, technical, and responsible public health for the development of practices that promote, audited and ensure the provision of radiological quality control and dosing in radiology service at national level. In view of the complexity and characteristics involved in relation to ionizing radiation, all assume their part in protecting the physical integrity of each user and a global perspective, to ensure the safeguarding of public health, while global and globalizing factor. To this end, and in the context of the publication of studies already carried out at European Union level, "European guidelines for quality criteria for computed tomography", it is proposed to establish guidelines in conducting studies to initially establish the comparison with the results obtained by QC and make adjustments if necessary, and subsequently implement a systematic periodic assessment frame that allows monitoring of data. Accordingly, it is proposed to conduct a national study involving the public network, private and University hospitals, that extends from the methodology used in previous studies in other countries of Europe, as is, selection of equipment of existing CT in Hospital Institution, where information will be gathered by filling out questionnaires concerning the equipment to be used. Data will be collected for the wearer, equipment and parameters of image acquisition, identifying diagnostic reference levels (NRD) in CT in Portuguese fact. A pilot study was carried out in an institution EPE and the values obtained are not significant, nor can they take predictive value given the small sample size. Despite this, suggest the existence of parameters that can be changed and this can vary the dose of radiation used It is hoped to get with this study, as mentioned, the basis of the establishment of NRD in CT in Portugal.

Generalized Extraction of Real-Time Parameters for Homogeneous Synchronous Dataflow Graphs

23rd Euromicro International Conference on Parallel, Distributed, and Network-Based Processing (PDP 2015). 4 to 6, Mar, 2015. Turku, Finland.

"Não nos deixem dormir...": Projeto de intervenção num Centro de Dia

O presente relatório apresenta o projeto desenvolvido na Casa-Acolhimento Santa Marta, cuja finalidade era a promoção de um envelhecimento ativo e bem-sucedido com vista à melhoria da qualidade de vida das pessoas idosas que frequentam a resposta social de Centro de Dia. O conhecimento coconstruído com as pessoas idosas e os profissionais da instituição permitiu a conceção e o desenvolvimento do projeto “Não nos deixem dormir…”. Sendo um projeto, elaborado em conjunto com os indivíduos, privilegiou os pressupostos da metodologia de investigação-ação participativa. Inerente a este posicionamento, incentivando a exploração e a rentabilização dos recursos e das potencialidades endógenas, bem como procurando atenuar ou resolver os problemas e as necessidades subjacentes, procurou-se tornar os sujeitos atores e autores das suas vidas. Deste modo, partindo dos contributos e das necessidades dos idosos o projeto justifica a sua importância, designadamente pela realização de ações que proporcionaram um maior número de atividades de acordo com as suas expectativas e os seus interesses e que promoveram as relações interpessoais propiciando momentos de convívio e de diálogo, fomentando o auto e o hétero conhecimento, bem como o respeito mútuo entre os idosos. De forma a sustentar a investigação e a intervenção, mobilizou-se contributos teóricos ligados sobretudo à terceira idade, que se tornaram essenciais na problematização, na reflexão e na ação. A concretização do projeto permitiu ainda uma constante reflexão acerca do papel do Educador Social junto da população idosa, bem como da pertinência da sua presença neste âmbito de intervenção.

Envelhecer: ser e estar - Projeto de intervenção num lar de idosos.

O presente documento reflete o percurso do desenho e do desenvolvimento de um projeto de Educação e Intervenção Social num Lar de idosos. Com base numa metodologia de Investigação-Ação Participativa, que privilegia os discursos, perceções e vivências dos atores sociais que integram esta realidade, procurou-se identificar os problemas e as necessidades. O conhecimento coconstruído com os idosos e profissionais da instituição permitiram a conceção e desenvolvimento do projeto “Envelhecer: ser e estar” que assentou, essencialmente, nas necessidades de (re) ativar e fortalecer as relações interpessoais, fomentar momentos de partilha, convívio, diálogo, discussão, negociação e participação, bem como, na urgência de envolver as pessoas na organização e ocupação do seu tempo livre promovendo a participação, o convívio e o lazer entre os idosos. Sendo um projeto das pessoas e com as pessoas, foram pensadas e planeadas ações e atividades que dessem resposta aos objetivos a que o projeto se propunha na tentativa de transformar e mudar os problemas e necessidades identificados pelos participantes. O projeto possibilitou, desta forma, iniciar um processo que permitiu fortalecer as relações entre os idosos, bem como envolvê-los na organização e participação das atividades do seu tempo livre, resultando num processo de transformação pessoal, grupal e institucional de permanente e contínua construção

Heróis de palmo e meio: a intervenção psicossocial e a relação de ajuda no acolhimento institucional

Os Lares de Infância e Juventude têm como objetivo defender o superior interesse das crianças, desempenhando um papel fundamental no acompanhamento do seu desenvolvimento. Este dever concretiza-se substancialmente na definição e acompanhamento dos seus projetos de vida. Neste sentido, as práticas dos profissionais devem ir ao encontro das reais necessidades dos seus atores, numa ação adequada, desenhada e concertada, tendo sempre como principio básico o respeito pelas idiossincrasias de cada criança e jovem. O presente Relatório tem como objetivo apresentar o desenho e desenvolvimento de um projeto de intervenção psicossocial e relação de ajuda com crianças em acolhimento institucional, intitulado “Heróis de Palmo e Meio. A intervenção psicossocial e a relação de ajuda no acolhimento institucional”. Este projeto teve como grande finalidade a “Participação das crianças e das suas famílias na construção dos seus projetos de vida com base na compreensão da institucionalização”. Priorizando as necessidades e respeitando o critério de urgência, foram desenhados dois subprojectos com duas crianças e suas famílias, tentando sempre responder às suas necessidades individuais. Foram definidos alguns objetivos gerais comuns, assim como objetivos específicos, estratégias e ações adequadas a cada um dos subprojectos. Os resultados deste projeto revelam-se significativos, já que permitiram, quer às crianças, quer às famílias, desenvolver um processo de reflexão e consciencialização em torno da sua história de vida, compreensão da sua realidade e participação ativa no projeto de vida dos filhos, o que veio facilitar o caminho inicial do processo da mudança.

Relatório de estágio em educação pré-escolar e ensino do 1º ciclo do ensino básico

O presente relatório – requisito parcial para obtenção do grau de mestre em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino Básico – reflete as experiências e conhecimentos desenvolvidos, resultantes da prática pedagógica supervisionada desenvolvida em dois contextos de estágio: Educação Pré-Escolar e 1.º Ciclo do Ensino Básico. Com efeito, a estudante teve como objetivo descrever, compreender e refletir acerca do processo de prática segundo o desenvolvimento de capacidades e competências substanciais à prática docente. Enquanto (futura) profissional de educação, verifica-se pertinente a mobilização de saberes científicos, pedagógicos e culturais, adquiridos ao longo da formação inicial para que se torne exequível uma prática sustentada. Similarmente, o docente, baseando-se em quadros teóricos e concetuais (amplificados de forma subjetiva e continuada), desenvolverá a sua forma pessoal de pensar e agir nos contextos de práticas reais visando a inclusão e equidade educativas, e a colaboração profissional e reflexiva. Conforme a metodologia de investigação-ação (constituída por várias etapas interligadas – observação, planificação, ação e avaliação, reflexão), a ação da mestranda desenvolveu-se de forma cíclica e articulada, numa perspetiva construtivista – e holística – do conhecimento a erigir pelas crianças (atores centrais do processo) ressalvando-se, igualmente, a pertinência dos demais instrumentos orientadores elaborados no decorrer da ação. Concludentemente, os estágios desenvolvidos nos dois contextos facultaram a edificação de uma postura profissional, reflexiva e investigativa, promotora da tomada de decisões em contexto de prática reafirmando-se competências profissionais e pessoais e valorando-se, efetivamente, a formação ao longo da vida para aquele que se constitui um docente generalista.

Cysticercus antibodies and antigens in serum from blood donors from Pondicherry, India

The aim of the present study was to screen the serum of blood donors, which are apparently healthy and residing in Pondicherry or its neighboring districts of Tamil Nadu State, for specific detection of Cysticercus antigens and antibodies. A total of 216 blood samples were collected from blood donors at the Central Blood Bank, JIPMER Hospital, Pondicherry, India during January and February 2004. Enzyme-linked immunosorbent assay (ELISA) was used to demonstrate anti-Cysticercus antibodies and the Co-agglutination (CoA) was used to detect antigen in sera. 14 (6.48 %) males were positive for either anti-Cysticercus antibodies or antigens. Of these eight sera were positive for anti-Cysticercus antibodies and six were positive for antigens. Results of the present study show that serum Cysticercus antigen detection may be a useful adjunct to antibody testing for seroprevalence studies of cysticercosis in the community. The present study is the first kind of study, carried out to determine both cysticercal antibodies as well as antigens in the serum samples collected from the healthy blood donors.

Innovations in technology: a friendly math project and the learning analytics “challenge”

Currently the world around us "reboots" every minute and “staying at the forefront” seems to be a very arduous task. The continuous and “speeded” progress of society requires, from all the actors, a dynamic and efficient attitude both in terms progress monitoring and moving adaptation. With regard to education, no matter how updated we are in relation to the contents, the didactic strategies and technological resources, we are inevitably compelled to adapt to new paradigms and rethink the traditional teaching methods. It is in this context that the contribution of e-learning platforms arises. Here teachers and students have at their disposal new ways to enhance the teaching and learning process, and these platforms are seen, at the present time, as significant virtual teaching and learning supporting environments. This paper presents a Project and attempts to illustrate the potential that new technologies present as a “backing” tool in different stages of teaching and learning at different levels and areas of knowledge, particularly in Mathematics. We intend to promote a constructive discussion moment, exposing our actual perception - that the use of the Learning Management System Moodle, by Higher Education teachers, as supplementary teaching-learning environment for virtual classroom sessions can contribute for greater efficiency and effectiveness of teaching practice and to improve student achievement. Regarding the Learning analytics experience we will present a few results obtained with some assessment Learning Analytics tools, where we profoundly felt that the assessment of students’ performance in online learning environments is a challenging and demanding task.

The International Multilateral System: Imperatives of Change, Potential Scenarios and Suggestions for the Future

Thesis presented to satisfy the necessary requirements for obtaining a PhD degree in International Relation with specialization in Globalization and the Environment,