Relatório de estágio apresentado à Universidade da Madeira para obtenção do grau de mestre em Ciências da Educação-Educação Sénior

O presente relatório refere-se ao estágio realizado no Lar Vila Assunção da Fundação Mary Jane Wilson, em São Gonçalo, na Região Autónoma da Madeira. O estágio é a unidade curricular que conclui o Mestrado em Ciências da Educação-Educação Sénior da Universidade da Madeira. O Lar Vila Assunção é uma IPSS (Instituição Particular de Solidariedade Social), direcionado para as pessoas idosas do sexo feminino. Neste contexto educacional (Lar), numa fase preliminar ou diagnóstica realizámos atividades em diversas áreas, temáticas e domínios, bem como em diferentes espaços de ação, no sentido de observarmos as capacidades e potencialidades dos idosos. Além das atividades, aplicámos um questionário com o objetivo de conhecer as características e os interesses de cada um dos idosos. Estes dados, no seu conjunto, permitiu estruturar o projeto de estágio propriamente dito. Após as reflexões sobre as atividades e a análise da recolha de dados do questionário, elaborámos um Projeto que se dividiu em duas áreas ou sub-projetos: “Ler + Cultura” e “Prática de Atividade Física”. Foi necessário fazer uma revisão de literatura sobre a temática do envelhecimento e as áreas específicas de forma a aprofundarmos os conteúdos significativos. Usámos em ambos os sub-projetos uma metodologia colaborativa e cooperativa que facilitou a entreajuda dos idosos, que naturalmente se apresentavam com níveis de envelhecimento diferentes. Esperemos que o conhecimento adquirido neste estágio possa contribuir para futuras práticas/reflexões e pesquisas sobre o idoso institucionalizado, refletindo-se, desta forma, na melhoria da sua qualidade de vida.

Relatório de estágio: educação pré-escolar e 1º CEB

No contexto do curso do 2º ciclo de Educação Pré-Escolar e Ensino e do 1º ciclo do Ensino Básico da Universidade da Madeira, realizou-se o presente relatório final, resultante da Unidade Curricular de Estágio e Relatório. Este relatório constitui a sistematização e reflexão escrita de todas as atividades e projetos realizados durante a intervenção pedagógica referente à valência de Educação Pré-Escolar e à do 1º Ciclo do Ensino Básico. Para além disso, também apresenta uma avaliação geral dos grupos de intervenção, como de uma criança, na vertente de Educação Pré-Escolar, no que se refere ao interesse e à participação nas propostas de trabalho.

Educação do campo e inovação pedagógica: perspectivas e possibilidades

As reflexões trazidas neste documento é fruto de uma longa estada no povoado de Lençóis na Escola Chico Mendes em Una-Bahia-Brasil, visando compreender quais ações pedagógicas estão sendo utilizadas para organização do conhecimento científico e social, o qual valoriza a aprendizagem do sujeito do campo. Dessa forma, para alcançar os objetivos previstos realizou-se a pesquisa à luz da etnografia, dando flexibilidade ao trabalho, construindo os ajustes necessários durante a pesquisa no campo. Nesse contexto, para a realização das análises, optou-se pela observação participante, análise dos artefatos culturais, dos documentos oficiais, diário escolar, vídeos e registros fotográficos, além da análise do discurso construído pelos sujeitos sociais: alunos, professor e comunidade, estes assumiram grande significado para a pesquisa. Por isso, pretendeu-se com este estudo, compreender se a educação do campo favorece a materialização de prática pedagógica inovadora, permitindo a participação de todos, por meio da pedagogia da equidade, e de ações multissensorias. Os resultados permitem-nos afirmar que os atos produzidos nos diversos contextos da educação do campo desse povoado valorizam a relação desses sujeitos com a terra, a aprendizagem cultural, a sua expressividade, e a ação pedagógica conscientizada. Os referenciais aqui utilizados contribuíram significativamente para interpretar o fenômeno e sustentar as possibilidades de inovação pedagógica na escola.

Development of a novel microextraction by packed sorbent-based approach followed by ultrahigh pressure liquid chromatography as a powerful technique for quantification phenolic constituents of biological interest in wines

A novel analytical approach, based on a miniaturized extraction technique, the microextraction by packed sorbent (MEPS), followed by ultrahigh pressure liquid chromatography (UHPLC) separation combined with a photodiode array (PDA) detection, has been developed and validated for the quantitative determination of sixteen biologically active phenolic constituents of wine. In addition to performing routine experiments to establish the validity of the assay to internationally accepted criteria (linearity, sensitivity, selectivity, precision, accuracy), experiments are included to assess the effect of the important experimental parameters on the MEPS performance such as the type of sorbent material (C2, C8, C18, SIL, and M1), number of extraction cycles (extract-discard), elution volume, sample volume, and ethanol content, were studied. The optimal conditions of MEPS extraction were obtained using C8 sorbent and small sample volumes (250 μL) in five extraction cycle and in a short time period (about 5 min for the entire sample preparation step). The wine bioactive phenolics were eluted by 250 μL of the mixture containing 95% methanol and 5% water, and the separation was carried out on a HSS T3 analytical column (100 mm × 2.1 mm, 1.8 μm particle size) using a binary mobile phase composed of aqueous 0.1% formic acid (eluent A) and methanol (eluent B) in the gradient elution mode (10 min of total analysis). The method gave satisfactory results in terms of linearity with r2-values > 0.9986 within the established concentration range. The LOD varied from 85 ng mL−1 (ferulic acid) to 0.32 μg mL−1 ((+)-catechin), whereas the LOQ values from 0.028 μg mL−1 (ferulic acid) to 1.08 μg mL−1 ((+)-catechin). Typical recoveries ranged between 81.1 and 99.6% for red wines and between 77.1 and 99.3% for white wines, with relative standard deviations (RSD) no larger than 10%. The extraction yields of the MEPSC8/UHPLC–PDA methodology were found between 78.1 (syringic acid) and 99.6% (o-coumaric acid) for red wines and between 76.2 and 99.1% for white wines. The inter-day precision, expressed as the relative standard deviation (RSD%), varied between 0.2% (p-coumaric and o-coumaric acids) and 7.5% (gentisic acid) while the intra-day precision between 0.2% (o-coumaric and cinnamic acids) and 4.7% (gallic acid and (−)-epicatechin). On the basis of analytical validation, it is shown that the MEPSC8/UHPLC–PDA methodology proves to be an improved, reliable, and ultra-fast approach for wine bioactive phenolics analysis, because of its capability for determining simultaneously in a single chromatographic run several bioactive metabolites with high sensitivity, selectivity and resolving power within only 10 min. Preliminary studies have been carried out on 34 real whole wine samples, in order to assess the performance of the described procedure. The new approach offers decreased sample preparation and analysis time, and moreover is cheaper, more environmentally friendly and easier to perform as compared to traditional methodologies.

Effectiveness of high-throughput miniaturized sorbent- and solid phase microextraction techniques combined with gas chromatography–mass spectrometry analysis for a rapid screening of volatile and semi-volatile composition of wines: a comparative study

In this study the feasibility of different extraction procedures was evaluated in order to test their potential for the extraction of the volatile (VOCs) and semi-volatile constituents (SVOCs) from wines. In this sense, and before they could be analysed by gas chromatography–quadrupole first stage masss spectrometry (GC–qMS), three different high-throughput miniaturized (ad)sorptive extraction techniques, based on solid phase extraction (SPE), microextraction by packed sorbents (MEPS) and solid phase microextraction (SPME), were studied for the first time together, for the extraction step. To achieve the most complete volatile and semi-volatile signature, distinct SPE (LiChrolut EN, Poropak Q, Styrene-Divinylbenzene and Amberlite XAD-2) and MEPS (C2, C8, C18, Silica and M1 (mixed C8-SCX)) sorbent materials, and different SPME fibre coatings (PA, PDMS, PEG, DVB/CAR/PDMS, PDMS/DVB, and CAR/PDMS), were tested and compared. All the extraction techniques were followed by GC–qMS analysis, which allowed the identification of up to 103 VOCs and SVOCs, distributed by distinct chemical families: higher alcohols, esters, fatty acids, carbonyl compounds and furan compounds. Mass spectra, standard compounds and retention index were used for identification purposes. SPE technique, using LiChrolut EN as sorbent (SPELiChrolut EN), was the most efficient method allowing for the identification of 78 VOCs and SVOCs, 63 and 19 more than MEPS and SPME techniques, respectively. In MEPS technique the best results in terms of number of extractable/identified compounds and total peak areas of volatile and semi-volatile fraction, were obtained by using C8 resin whereas DVB/CAR/PDMS was revealed the most efficient SPME coating to extract VOCs and SVOCs from Bual wine. Diethyl malate (18.8 ± 3.2%) was the main component found in wine SPELiChrolut EN extracts followed by ethyl succinate (13.5 ± 5.3%), 3-methyl-1-butanol (13.2 ± 1.7%), and 2-phenylethanol (11.2 ± 9.9%), while in SPMEDVB/CAR/PDMS technique 3-methyl-1-butanol (43.3 ± 0.6%) followed by diethyl succinate (18.9 ± 1.6%), and 2-furfural (10.4 ± 0.4%), are the major compounds. The major VOCs and SVOCs isolated by MEPSC8 were 3-methyl-1-butanol (26.8 ± 0.6%, from wine total volatile fraction), diethyl succinate (24.9 ± 0.8%), and diethyl malate (16.3 ± 0.9%). Regardless of the extraction technique, the highest extraction efficiency corresponds to esters and higher alcohols and the lowest to fatty acids. Despite some drawbacks associated with the SPE procedure such as the use of organic solvents, the time-consuming and tedious sampling procedure, it was observed that SPELiChrolut EN, revealed to be the most effective technique allowing the extraction of a higher number of compounds (78) rather than the other extraction techniques studied.

An improved and fast UHPLC-PDA methodology for determination of L-ascorbic and dehydroascorbic acids in fruits and vegetables: evaluation of degradation rate during storage

This study provides a versatile validated method to determine the total vitamin C content, as the sum of the contents of L-ascorbic acid (L-AA) and dehydroascorbic acid (DHAA), in several fruits and vegetables and its degradability with storage time. Seven horticultural crops from two different origins were analyzed using an ultrahigh-performance liquid chromatographic–photodiode array (UHPLC-PDA) system, equipped with a new trifunctional high strength silica (100% silica particle) analytical column (100 mm×2.1 mm, 1.7 μm particle size) using 0.1% (v/v) formic acid as mobile phase, in isocratic mode. This new stationary phase, specially designed for polar compounds, overcomes the problems normally encountered in HPLC and is suitable for the analysis of large batches of samples without L-AA degradation. In addition, it proves to be an excellent alternative to conventional C18 columns for the determination of L-AA in fruits and vegetables. The method was fully validated in terms of linearity, detection (LOD) and quantification (LOQ) limits, accuracy, and inter/intraday precision. Validation experiments revealed very good recovery rate of 96.6±4.4% for L-AA and 103.1±4.8 % for total vitamin C, good linearity with r2-values >0.999 within the established concentration range, excellent repeatability (0.5%), and reproducibility (1.6%) values. The LOD of the method was 22 ng/mL whereas the LOQ was 67 ng/mL. It was possible to demonstrate that L-AA and DHAA concentrations in the different horticulture products varied oppositely with time of storage not always affecting the total amount of vitamin C during shelf-life. Locally produced fruits have higher concentrations of vitamin C, compared with imported ones, but vegetables showed the opposite trend. Moreover, this UHPLC-PDA methodology proves to be an improved, simple, and fast approach for determining the total content of vitamin C in various food commodities, with high sensitivity, selectivity, and resolving power within 3 min of run analysis.

Relatório de estágio de mestrado

Este relatório foi elaborado para obtenção do grau de mestre em Educação PréEscolar e Ensino do 1.º Ciclo do Ensino Básico. Retrata algumas das experiências vividas na valência a educação pré-escolar aquando do estágio na Escola Básica do 1º Ciclo com Pré-Escolar da Achada. Uma vez que quem faz da educação o seu campo de ação não pode ter uma mente "vazia", toda a prática pedagógica foi norteada por pressupostos teóricos e metodológicos, sendo que em todos eles não foi esquecida a maneira participativa com que olho a pedagogia. Conceitos como a autonomia, a cooperação, a participação ativa da criança e o envolvimento da família no processo educativo foram fundamentais, na medida em que auxiliaram e enriqueceram o planeamento de todo o trajeto desenvolvido. O educador vive constantemente momentos de reflexão, que visam o aperfeiçoamento da sua intervenção, sendo a investigação-ação uma metodologia auxiliar nesse processo. Ótima como forma de formação ao longo da vida, porque permite "ver" para além da prática e da ação da criança, permitiu-me traçar planos interventivos com o objetivo de responder a algumas questões problemáticas observadas. De realçar que o docente vive um dia de cada vez e deve ser capaz de responder às necessidades, interesses e sugestões das crianças, colocando-as em primeiro plano. Deste modo, o presente relatório explana algumas das atividades desenvolvidas nesta etapa de formação académica que me permitiu aprender a aprender a ser educadora, porque esta caminhada não é o fim de uma formação docente, apenas me abre as portas a novos momentos de aprendizagem.

A micro-extraction technique using a new digitally controlled syringe combined with UHPLC for assessment of urinary biomarkers of oxidatively damaged DNA

The formation of reactive oxygen species (ROS) within cells causes damage to biomolecules, including membrane lipids, DNA, proteins and sugars. An important type of oxidative damage is DNA base hydroxylation which leads to the formation of 8-oxo-7,8-dihydro-29-deoxyguanosine (8-oxodG) and 5-hydroxymethyluracil (5-HMUra). Measurement of these biomarkers in urine is challenging, due to the low levels of the analytes and the matrix complexity. In order to simultaneously quantify 8-oxodG and 5-HMUra in human urine, a new, reliable and powerful strategy was optimised and validated. It is based on a semi-automatic microextraction by packed sorbent (MEPS) technique, using a new digitally controlled syringe (eVolH), to enhance the extraction efficiency of the target metabolites, followed by a fast and sensitive ultrahigh pressure liquid chromatography (UHPLC). The optimal methodological conditions involve loading of 250 mL urine sample (1:10 dilution) through a C8 sorbent in a MEPS syringe placed in the semi-automatic eVolH syringe followed by elution using 90 mL of 20% methanol in 0.01% formic acid solution. The obtained extract is directly analysed in the UHPLC system using a binary mobile phase composed of aqueous 0.1% formic acid and methanol in the isocratic elution mode (3.5 min total analysis time). The method was validated in terms of selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), extraction yield, accuracy, precision and matrix effect. Satisfactory results were obtained in terms of linearity (r2 . 0.991) within the established concentration range. The LOD varied from 0.00005 to 0.04 mg mL21 and the LOQ from 0.00023 to 0.13 mg mL21. The extraction yields were between 80.1 and 82.2 %, while inter-day precision (n=3 days) varied between 4.9 and 7.7 % and intra-day precision between 1.0 and 8.3 %. This approach presents as main advantages the ability to easily collect and store urine samples for further processing and the high sensitivity, reproducibility, and robustness of eVolHMEPS combined with UHPLC analysis, thus retrieving a fast and reliable assessment of oxidatively damaged DNA.