A biomimetic sensor for monitoring oxidative stress biomarker in point-of-care

1st ASPIC International Congress

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.

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.

Voltammetric immunosensor for the diagnosis of celiac disease based on the quantification of anti-gliadin antibodies

Antibodies against gliadin are used to detect celiac disease (CD) in patients. An electrochemical immunosensor for the voltammetric detection of human anti-gliadin antibodies (AGA) IgA and AGA IgG in real serum samples is proposed. The transducer surface consists of screen-printed carbon electrodes modified with a carbon nanotube/gold nanoparticle hybrid system, which provides a very useful surface for the amplification of the immunological interactions. The immunosensing strategy is based on the immobilization of gliadin, the antigen for the autoantibodies of interest, onto the nanostructured surface. The antigen–antibody interaction is recorded using alkaline phosphatase labeled anti-human antibodies and a mixture of 3-indoxyl phosphate with silver ions (3-IP/Ag+) was used as the substrate. The analytical signal is based on the anodic redissolution of the enzymatically generated silver by cyclic voltammetry. The electrochemical behavior of this immunosensor was carefully evaluated assessing aspects as sensitivity, non-specific binding and matrix effects, and repeatability and reproducibility. The results were supported with a commercial ELISA test.

Multiplexed electrochemical immunosensor for detection of celiac disease serological markers

Celiac disease (CD) is a gluten-induced autoimmune enteropathy characterized by the presence of antibodies against gliadin (AGA) and anti-tissue transglutaminase (anti-tTG) antibodies. A disposable electrochemical dual immunosensor for the simultaneous detection of IgA and IgG type AGA and antitTG antibodies in real patient’s samples is presented. The proposed immunosensor is based on a dual screen-printed carbon electrode, with two working electrodes, nanostructured with a carbon–metal hybrid system that worked as the transducer surface. The immunosensing strategy consisted of the immobilization of gliadin and tTG (i.e. CD specific antigens) on the nanostructured electrode surface. The electrochemical detection of the human antibodies present in the assayed serum samples was carried out through the antigen–antibody interaction and recorded using alkaline phosphatase labelled anti-human antibodies and a mixture of 3-indoxyl phosphate with silver ions was used as the substrate. The analytical signal was based on the anodic redissolution of enzymatically generated silver by cyclic voltammetry. The results obtained were corroborated with commercial ELISA kits indicating that the developed sensor can be a good alternative to the traditional methods allowing a decentralization of the analyses towards a point-of-care strategy.

Orchestration of e-learning services for automatic evaluation of programming exercises

Managing programming exercises require several heterogeneous systems such as evaluation engines, learning objects repositories and exercise resolution environments. The coordination of networks of such disparate systems is rather complex. These tools would be too specific to incorporate in an e-Learning platform. Even if they could be provided as pluggable components, the burden of maintaining them would be prohibitive to institutions with few courses in those domains. This work presents a standard based approach for the coordination of a network of e-Learning systems participating on the automatic evaluation of programming exercises. The proposed approach uses a pivot component to orchestrate the interaction among all the systems using communication standards. This approach was validated through its effective use on classroom and we present some preliminary results.

Satisfação profissional dos enfermeiros em unidades de cuidados continuados integrados

A Satisfação no trabalho, definida como reações individuais, cognitivas, afetivas e avaliativas perante o trabalho (Locke, 1983), tem sido amplamente estudada no âmbito das organizações. A sua medição na área da saúde contribui para incrementar um bom ambiente na prática e bem-estar dos profissionais e para aumentar a qualidade dos cuidados. Em 2006 surge um novo contexto de trabalho através da criação da RNCCI, que pretende fazer face ao crescente aumento de população em situação de dependência ou incapacidade, mas também aliviar a sobrecarga e custos associados aos cuidados hospitalares. A reorganização do trabalho nas unidades de cuidados continuados acenta nos princípios da interdisciplinariedade e cooperação entre a equipa, requerendo mudança e adaptação à prática. Sendo os enfermeiros a classe profissional com maior tempo de contacto nos cuidados, importa verificar se estão satisfeitos com a mudança induzida. Assim, este estudo de natureza quantitativa pretende medir e analisar a satisfação profissional ddos enfermeiros que trabalham no serviço de longa duração e manutenção nas unidades de cuidados continuados do distrito de Braga, e verificar se esta variável é influenciada por fatores sociodemográficos e laborais. Para a recolha de dados utiliza-se a Escala de Avaliação da Satisfação no Trabalho para Enfermeiros - EASPE© (α=0,814) e um questionário sociodemográfico e profissional. A análise dos dados é efetuada com recurso à estatística descritiva e inferencial com utilização dos testes de Mann-Whitney e de Kruskal-Wallis. Os resultados obtidos evidenciam que os enfermeiros apresentam um nível de satisfação global no trabalho positivo, mas negativo nas dimensões satisfação com benefícios e recompensas e satisfação com promoção. Conclui-se, ainda, que a satisfação com promoção é influenciada pelas habilitações literárias, que a satisfação com a comunicação é influenciada pela remuneração, que a satisfação com benefícios e recompensas é influenciada pelo horário semanal e que a unidade de cuidados é determinante na satisfação global no trabalho.