Browsing by Issue Date, starting with "2022-11"
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- Green extraction approach based on μSPEed® followed by HPLC-MS/MS for the determination of atropine and scopolamine in tea and herbal tea infusionsPublication . González-Gómez, Lorena; Pereira, Jorge A. M.; Morante-Zarcero, Sonia; Câmara, José S.; Sierra, Isabel; Augusto Machado Pereira, Jorge; Câmara, Joséliquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been proposed for the analysis of atropine and scopolamine in tea and herbal tea infusions. For this, a digiVOL® Digital Syringe was used with different sorbents and working conditions to obtain a fast and efficient µSPEed® extraction. The best performance was achieved with a PS/DVB sorbent phase, sample loading of 5 × 500 µL and elution with 2 × 100 µL aliquots of methanol. The strategy based on µSPEed® followed by HPLC-MS/MS was validated, attaining quantitation limits lower than 0.15 ng mL− 1 and recoveries between 94 and 106% for both analytes and applied to seventeen tea and herbal tea infusions. Fourteen infusions showed contamination with one or both analytes above the maximum content legislated (sum of atropine and scopolamine < 0.2 ng mL− 1).
- Relatório de estágio para obtenção do grau de mestre em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino BásicoPublication . Gomes, Ana Cristina Azevedo; Correia, Fernando Luís de SousaO presente relatório de estágio surge no âmbito do Mestrado em Educação Pré Escolar e Ensino do 1.º Ciclo do Ensino Básico e tem como finalidade a obtenção do grau de mestre. Neste documento são apresentadas descrições detalhadas e reflexões acerca das três Práticas Pedagógicas (PP) vivenciadas, nomeadamente a PP I, que se desenvolveu na Escola Básica do 1.º Ciclo com Pré-Escolar da Achada, com o grupo da Sala do Arco Íris e as PP II e III, que se desenvolveram na Escola Básica do 1.º Ciclo com Pré-Escolar da Ladeira, com as turmas 4.º A e 4.º B. Saliento ainda que, a prática com a turma 4.º A se desenvolveu em ensino a distância e que, por sua vez, a prática com a turma 4.º B se desenrolou em ensino presencial. No que diz respeito à estrutura, numa primeira etapa são evidenciadas quais as bases teóricas e metodológicas que fundamentaram as práticas, sendo que numa segunda etapa são apresentados diversos momentos de aprendizagem desenvolvidos com as crianças, bem como algumas reflexões. Com o objetivo de colmatar as problemáticas identificadas nos diferentes grupos, foram desenvolvidos projetos de Investigação-Ação, que tiveram como ponto de partida as seguintes questões: “Como podem as crianças da Sala do Arco-Íris desenvolver noções matemáticas?” (PP I); “Como podem os alunos do 4.º B desenvolver atitudes de saber estar em sala de aula?” (PP III). É importante salientar ainda que, para dar respostas às questões apresentadas foi necessário efetuar uma pesquisa aprofundada sobre a metodologia de Investigação-Ação. Em suma, este relatório é uma compilação de tudo o que foi aprendido e vivenciado ao longo do meu percurso académico, mais em específico durante os dois anos de mestrado.
- Relatório de estágio para a obtenção do grau de mestre em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino BásicoPublication . Alves, Micaela Arnek Rodrigues da Costa; Camacho, Maria José de JesusO presente relatório de estágio surge no âmbito do Mestrado em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino Básico e possui como principal objetivo a obtenção de grau de mestre nestas áreas de educação. Este tem como finalidade a partilha das experiências vivenciadas ao longo das três práticas pedagógicas desenvolvidas nos contextos educativos enunciados anteriormente, sendo que a prática referente ao Pré-Escolar decorreu no Auxílio Maternal do Funchal e as práticas do 1.º Ciclo do Ensino Básico no Colégio do Marítimo. Cada vez mais compreende-se que o processo educativo deve apresentar na sua essência a criança, sendo esta responsável pela aquisição e desenvolvimento de novos conhecimentos. Para que tal possa acontecer, deve dar-se maior relevância à Inovação Pedagógica a fim de colmatar as consequências de uma educação tradicional. Numa primeira parte deste relatório apresenta-se uma fundamentação teórica que contextualiza as práticas evidenciadas, na medida em que toda a ação educativa deverá possuir como alicerce uma intencionalidade pedagógica e didática. De modo a alcançar uma atitude reflexiva e crítica e com o intuito de resolver problemáticas específicas de cada grupo de intervenção, desenvolveram-se três projetos baseados na metodologia de Investigação ação. Assim sendo, na parte dois deste relatório apresenta-se uma breve fundamentação que sustenta os projetos desenvolvidos. Por fim, a terceira parte deste relatório destinar-se-á à explanação das práticas desenvolvidas nos contextos referenciados anteriormente. Em suma, este relatório constitui-se como uma profunda reflexão e análise de toda a prática desenvolvida ao longo da minha formação, tendo por base uma revisão literária que possui como principal objetivo colocar em primeiro lugar os interesses e as necessidades das crianças.
- Bioactive compounds and antioxidant activity from spent coffee grounds as a powerful approach for Its valorizationPublication . Andrade, Carolina; Perestrelo, Rosa; Câmara, José S.Coffee is one of the world’s most popular beverages, and its consumption generates copious amounts of waste. The most relevant by-product of the coffee industry is the spent coffee grounds, with 6 million tons being produced worldwide per year. Although generally treated as waste, spent coffee grounds are a rich source of several bioactive compounds with applications in diverse industrial fields. The present work aimed at the analysis of spent coffee grounds from different geographical origins (Guatemala, Colombia, Brazil, Timor, and Ethiopia) for the identification of bioactive compounds with industrial interest. For this purpose, the identification and quantification of the bioactive compounds responsible for the antioxidant activity attributed to the spent coffee grounds were attempted using miniaturized solid-phase extraction (µ-SPEed), combined with ultrahigh-performance liquid chromatography with photodiode array detection (UHPLC-PDA). After validation of the µ-SPEed/UHPLC-PDA method, this allowed us to conclude that caffeine and 5-caffeoylquinic acid (5-CQA) are the most abundant bioactive compounds in all samples studied. The total phenolic content (TPC) and antioxidant activity are highest in Brazilian samples. The results obtained show that spent coffee grounds are a rich source of bioactive compounds, supporting its bioprospection based on the circular economy concept closing the loop of the coffee value chain, toward the valorization of coffee by-products.
- Relatório de estágio para a obtenção do grau de mestre em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino BásicoPublication . Tanque, Tanya Isabel Azevedo; Gouveia, Ana IsabelO presente relatório espelha todo o processo de aquisição de competências e tem como principal objetivo refletir e partilhar as experiências vividas durante este percurso de formação inicial para a docência nas valências de Educação Pré-Escolar e do 1.º Ciclo do Ensino Básico. Organizado em dois momentos que se complementam, este relatório visa apresentar na primeira parte as bases teóricas e metodológicas que fundamentam as Práticas Pedagógicas que, por sua vez, surgem na segunda parte. Relacionado com as Práticas Pedagógicas torna-se importante referir que se adotou, aquando das mesmas, uma postura crítico-reflexiva fundamental para a identificação de problemáticas. Após a identificação das problemáticas em ambas as valências, procurou se desenvolver através da metodologia de Investigação-Ação dois projetos que se basearam nas seguintes questões-problema: (1) Educação Pré-Escolar: Como é que as crianças da Sala da Pré B poderão desenvolver atitudes de formação pessoal e social nomeadamente de cumprimento de regras? (2) 1º Ciclo do Ensino Básico: Como auxiliar os alunos do 2.ºB a aprimorar as suas competências ao nível da leitura e da escrita? É de salientar que este documento é, portanto, o testemunho de um percurso de formação inicial para a docência, onde se pretende explanar todas as aprendizagens adquiridas, sendo das mais importantes, observar, refletir, planificar e atuar, tendo em consideração os interesses e as dificuldades da criança, colocando-a no centro de todo o processo de ensino-aprendizagem.
- Spheroid development for in vitro Chondrogenesis: preliminary studiesPublication . Jardim, Filipa Raquel Nunes; Tomás, Helena Maria Pires Gaspar; Gonçalves, Mara Isabel JesusThis thesis presents the first set of experimental work done at CQM – Centro de Química da Madeira aimed at the implementation of a new research line focused on spheroids for chondrogenesis study. Three different techniques for spheroid preparation using human mesenchymal stem cells (hMSCs) were tested. The preparation of spheroids by cell self-aggregation in ultra-low adhesion (ULA) cell culture plates was shown to be easy to use, and to lead to stable spheroids. The addition of 50 ng/mL carboxymethyl cellulose (CMC) to the cell culture medium helped to control spheroid’s size and spheroidicity, improving cell viability as well. The influence of different culture medium compositions on cell metabolic activity and on cell differentiation towards the chondrogenic lineage was evaluated in 2D cell cultures. The addition of TGF-β1, as well as other supplements, to the chondrogenic medium (CM) increased cell proliferation, and induced chondrogenesis (revealed through histochemical staining). For the assessment of the chondrogenesis in 3D constructs, spheroids were initially prepared and matured for 14 days (50 ng/mL of CMC was added to the medium at day 5) in ULA plates (poly-HEMA coating) in BM with high glucose content, at a high cell seeding density. Then, a spheroid group was exposed to CM; and another maintained in Basal medium (BM), as a control (CTRL) group for more 28 days (chondrogenesis induction). Spheroid’s growth was observed in the CM group, whereas their shrinking was evident in CTRL group. Also, CM led to higher cell viability values in spheroids. Histochemistry staining experiments were only successful using the Fast Green/Safranin O system, revealing the occurrence of chondrogenesis in spheroids exposed to CM. ALP activity measurements showed the absence of the hypertrophic phenotype in the differentiated spheroids (chondrospheres). Finally, ELISA assays allowed the detection of collagen type II and aggrecan in the chondrospheres cultured in CM.
- Urinary metabolomic fingerprints as a powerful tool for the next generation of prostate cancer diagnosisPublication . Berenguer, Cristina Viveiros; Câmara, José de Sousa; Pereira, Jorge Augusto MachadoProstate cancer (PCa) remains the most frequent malignant tumour and a leading cause of oncological death in men, despite the spectacular advances in molecular medicine, including genomics, proteomics, transcriptomics, lipidomics, and personalized medicine. Apart from classical biomarkers, the study of endogenous volatile organic metabolites (VOMs) biosynthesized by different metabolic pathways and found in several biofluids is emerging as an innovative, efficient, and non-invasive source of data to establish the volatilomic biosignature of PCa. In this context, the primary objective of this thesis was to establish the urinary volatilomic profile of PCa using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME/GC-MS). This non-invasive approach to set putative PCa biomarkers was applied to PCa patients (n = 29), men subjected to a radical prostatectomy (RP, n = 34), and cancer-free individuals (control group, n = 49). A total of 60 VOMs belonging to different chemical families were identified in the groups under study. This included phenolic compounds, terpenes, norisoprenoids, ketones, alcohols, and sulphur containing and furanic compounds. The data matrix obtained was submitted to multivariant analysis, through partial least-squares discriminant analysis (PLS-DA). The results obtained did not show complete discrimination between the groups under study, since more than 50 % of the variability in the outcome data could not be explained by the models. The heatmap according to Pearson’s correlation showed that 2-(1-cyclopentyl)furan, 2-pentanone and 2-bromophenol were more associated with the control group, while carvone, α-corocalene, 2-acetylfuran, and cyclohexanone showed high correlations with the PCa group. In contrast, 2-ethyl-5- methylfuran, methanethiol, o-methoxyphenol, p-cymenene, and 3,4-dehydro-β-ionene, were more associated with the RP group. An exhaustive study of the demographic and clinical characteristics of the patients recruited is crucial to elucidate the changes that occur in their volatilomic profile. Moreover, a larger cohort of samples is necessary to improve the predictive power and reliability of the statistical models developed.
- Implicações do mobbing no bem-estar familiarPublication . Pereira, Estela Simone Ornelas; Garcês, Soraia Fernandes; Pocinho, Margarida Maria Ferreira Diogo DiasCom base na literatura, o Mobbing é caracterizado pela prática de comportamentos hostis e de longa duração no local de trabalho, colocando a vítima numa posição vulnerável e sem apoio e que pode levar esta a afastar-se ao máximo do seu ambiente de trabalho (Leymann, 1996). As redes de apoio são fundamentais para ultrapassar este tipo de desafios, sendo a principal delas a família que é o primeiro agente de socialização dos indivíduos (Krys et al., 2021). Neste sentido, esta investigação tem como objetivo perceber se a ocorrência de Mobbing tem impacto no bem-estar familiar dos docentes em Portugal. A amostra deste estudo contou com 406 participantes com idades compreendidas entre os 21 e 69 anos. Relativamente aos instrumentos utilizados, foi construída uma Escala de Bem-Estar Familiar com base no Modelo PERMA (Pereira, Garcês, & Pocinho, submetido) e a Escala de Avaliação do Mobbing (Pocinho & Garcês, 2018). No que concerne aos resultados, é possível verificar que existe uma correlação negativa entre o Mobbing e o Bem-estar familiar. Não existem diferenças significativas em relação ao género, idade, meio ou ciclo que lecionam. Possuir uma relação é a variável que apresenta resultados significativos, mostrando que quem se encontra numa relação apresenta menores níveis de Mobbing e maior Bem-estar familiar.
- Preparation and characterization of anionic PPI dendrimers for delivery of the hydroxychloroquine into tumor cellsPublication . Rodrigues, Ana Filipa Gonçalves; Rodrigues, João Manuel Cunha; Maciel, Dina Maria SousaCancer is one of the most lethal diseases worldwide. In 2020, 19.3 million new cancer cases were reported, with approximately 10 million deaths. Female breast cancer was the most diagnosed, with 2.3 million new cases. Hydroxychloroquine (HCQ) is an immunomodulatory drug used to treat malaria and has been shown to have therapeutic potential in oncology. It has been proven that HCQ increases tumor cell death alone or combined with targeted agents or cytotoxic chemotherapy. By combining HCQ and a nanocarrier, it is possible to direct the drug to the tumor tissues, reducing the side effects and enhancing the bioavailability. Based on the previous experience of our group, the main goal of this master thesis was to prepare anionic carboxylate and sulfonate PPI dendrimers to transport and deliver the HCQ into tumor cells. First, amine-terminated PPI dendrimers, generation 0 to 3 (G0-G3), were synthesized, with a yield in the range of 57% -99%, and characterized by 1H- and 13C- NMR. After the G0 to G3 amine-terminated PPI dendrimers were prepared and obtained, the functionalization of the terminal groups with carboxylate and sulfonate dendrimers was done. Additionally, for the first time, the carboxylate- and sulfonate-terminated dendrimers G4 was synthesized. The anionic dendrimers have the advantage of being less cytotoxic, hemotoxic, and immunogenicity than cationic dendrimers. After G1 to G3 characterization by 1H 13C NMR, ATR-FTIR, DLS, zeta potential, and MS, HCQ was encapsulated. Following this, the in vitro drug release was performed in PBS with a pH 5 and 7.4, since the cumulative release is slightly higher in PBS with a pH 5. In general, all dendrimers showed a sustained release of HCQ in both pHs. The hemotoxicity of the dendrimers and the HCQ were also evaluated and showed no hemotoxicity. Finally, the cytotoxicity of the synthesized dendrimers, non-loaded and loaded with HCQ, was evaluated in a cancer cell line (MCF-7) and a non-cancer cell line (BJ cells). The HCQ in a range of concentration between 0.5 and 50 µM are highly cytotoxic for BJ cells and less cytotoxic for MCF-7 cells. The HCQ was further tested with the CACO-2 cells and showed to be less sensitive to the drug than with the MCF-7 cells. In order to improve the cytotoxicity of the prepared dendrimers, an assay with doxorubicin (DOX) was performed, but no synergetic effect was observed.
- MALDI-TOF MS-based urinary proteomic/peptidomic signature of breast cancer as innovative diagnostic approachPublication . Sousa, Patrícia Maria Gonçalves; Gouveia, Rosa Maria de Sá Perestrelo; Câmara, José de SousaThe development of a rapid and high-sensitive breast cancer (BC) diagnostic method has been increasingly investigated by many researchers, to have a more effective therapy, through the early detection of this disease that affects millions of women worldwide. Therefore, the discovery of specific biomarkers is one of the topics in clinical medicine, however several challenges remain in the development of optimal sample preparation for proteomic/peptidomic analysis of urine, due to its highly variable content, as well as the presence of various proteins in low abundance or modified forms. Thus, this investigation aims to establish potential BC urinary protein biomarkers using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D SDS-PAGE) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOF MS) for BC diagnosis and monitoring the efficiency of therapy. The results of the Lowry´s assay demonstrated that total protein concentration increased after precipitation and that healthy control group (HCs) have higher total protein concentrations than BC patients. Related to MALDI-TOF MS analysis, results revealed that four protein/peptide ion signatures (m/z 1046.5, 1062.4, 1237.7 and 1727.9), with variable importance in projection (VIP) > 1, allowed the discrimination between BC patients and HCs. The discrimination efficiency and accuracy of BC urine proteins/peptides were ascertained by receiver operating characteristic (ROC) curve analysis that allowed the identification of some features with high sensitivity and specificity (99.6%). Therefore, the obtained data revealed MALDI-TOF MS as a powerful tool to explore proteomic/peptidomic biosignatures, due to its speed, sensitivity, and mass accuracy, which allow establishing novel disease biomarkers. Nevertheless, a deep study using a higher number of samples must be carried out to confirm and consolidate the data obtained.
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