Revisão do plano de segurança da água do sistema regional do Carvoeiro

Os Planos de Segurança da Água surgem com a necessidade de aumentar a segurança da água de abastecimento, superando a monitorização de conformidade de “fim de linha”, permitindo aumentar a confiança do consumidor na qualidade da água que lhe é fornecida. Esta nova abordagem recorre a uma metodologia de gestão baseada na identificação e no controlo de riscos em pontos críticos de um sistema de abastecimento, em complemento do controlo realizado através da monitorização da conformidade da água entregue aos consumidores. O Plano de Segurança da Água (PSA) encontra-se implementado no Sistema Regional do Carvoeiro (SRC) desde o ano de 2009. O SRC é um sistema de abastecimento de água em alta, sendo constituído por conjunto de infraestruturas de captação, tratamento, transporte e armazenamento de água desde a sua origem, localizada no rio Vouga, em Carvoeiro, até aos municípios integrados na Associação de Municípios do Carvoeiro-Vouga. Atendendo à obra de expansão do SRC, tornou-se imperativo efetuar uma revisão ao PSA, sendo este o objetivo primordial do trabalho de estágio desenvolvido na empresa Águas do Vouga S.A, concessionária responsável pela gestão do SRC. Para a prossecução deste objetivo, o trabalho desenvolvido envolveu os seguintes passos metodológicos: identificação das operações aplicadas no SRC; identificação de perigos e eventos perigosos em todos os órgãos constituintes do sistema; avaliação de riscos; identificação de pontos críticos de controlo; identificação de pontos de monitorização e medidas preventivas; elaboração do plano de monitorização, incluindo, procedimentos de controlo operacional em condições normais de funcionamento e em caso de desvio; validação deste plano. Deste trabalho resultou a identificação de 166 eventos perigosos, 17 tipologias de perigos, 3 pontos de controlo crítico e 17 pontos de monitorização. Os pontos de controlo crítico foram identificados nos processos de tratamento da ETA do Carvoeiro. O primeiro foi localizado na etapa de filtração com areia, antracite e zeólitos correspondendo aos perigos com metais (Fe e Mn), outros compostos químicos perigosos, partículas, turvação, matéria orgânica e alumínio. O segundo ponto foi identificado na etapa de filtração com filtros de carvão ativado granular relativo ao aparecimento de sabor e cianotoxinas. O terceiro ponto de controlo crítico foi encontrado na etapa de desinfeção referente aos microrganismos patogénicos. Os pontos de monitorização foram localizados ao longo do sistema em situações onde não se dispõem de nenhuma medida de controlo para eliminar o perigo e antes e após os pontos de controlo crítico. O plano de monitorização foi desenvolvido para estes pontos, embora os limites e procedimentos definidos devam ser alvo de revisão após a conclusão da obra de expansão do sistema. A validação da revisão do plano foi iniciada, mas cingiu-se apenas na avaliação preliminar de riscos, prévia ao início de operação da ETA do Carvoeiro. Para além da revisão deste plano, foram realizadas outras tarefas, nomeadamente uma análise à qualidade da água fornecida e distribuída pelo sistema, a elaboração do plano PCQA para o ano de 2016, a configuração da plataforma de gestão operacional NAVIATM e a revisão do Manual de Gestão da Águas do Vouga relativo ao processo de qualidade na captação, tratamento e distribuição e ao processo de qualidade na gestão do PSA.

Characterization of water-soluble organic compounds in bioaerosols by liquid chromatography and fluorescence spectroscopy

In the last decades, the effects of the air pollution have been increasing, especially in the case of the human health diseases. In order to overcome this problem, scientists have been studying the components of the air. As a part of water-soluble organic compounds, amino acids are present in the atmospheric environment as components of diverse living organisms which can be responsible for spreading diseases through the air. Liquid chromatography is one technique capable of distinguish the different amino acids from each other. In this work, aiming at separating the amino acids found in the aerosols samples collected in Aveiro, the ability of four columns (Mixed-Mode WAX-1, Mixed-Mode HILIC-1, Luna HILIC and Luna C18) to separate four amino acids (aspartic acid, lysine, glycine and tryptophan) and the way the interaction of the stationary phases of the columns with the analytes is influenced by organic solvent concentration and presence/concentration of the buffer, are being assessed. In the Mixed-Mode WAX-1 column, the chromatograms of the distinct amino acids revealed the separation was not efficient, since the retention times were very similar. In the case of lysine, in the elution with 80% (V/V) MeOH, the peaks appeared during the volume void. In the Mixed-Mode HILIC-1 column, the variation of the organic solvent concentration did not affect the elution of the four studied amino acids. Considering the Luna HILIC column, the retention times of the amino acids were too close to each other to ensure a separation among each other. Lastly, the Luna C18 column revealed to be useful to separate amino acids in a gradient mode, being the variation of the mobile phase composition in the organic solvent concentration (ACN). Luna C18 was the column used to separate the amino acids in the real samples and the mobile phase had acidified water and ACN. The gradient consisted in the following program: 0 – 2 min: 5% (V/V) ACN, 2 – 8 min: 5 – 2 % (V/V) ACN, 8 – 16 min: 2% (V/V) ACN, 16 – 20 min: 2 – 20 % (V/V) ACN, 20 – 35 min: 20 – 35 % (V/V) ACN. The aerosols samples were collected by using three passive samplers placed in two different locations in Aveiro and each sampler had two filters - one faced up and the other faced down. After the sampling, the water-soluble organic compounds was extracted by dissolution in ultra-pure water, sonication bath and filtration. The resulting filtered solutions were diluted in acidified water for the chromatographic separation. The results from liquid chromatography revealed the presence of the amino acids, although it was not possible to identify each one of them individually. The chromatograms and the fluorescence spectra showed the existence of some patterns: the samples that correspond to the up filters had more intense peaks and signals, revealing that the up filters collected more organic matter.

CD81 interacting proteins

Fertilization is a multistep and complex process culminating in the merge of gamete membranes, cytoplasmic unity and fusion of genome. CD81 is a tetraspanin protein that participates in sperm-oocyte interaction, being present at the oocyte surface. CD81 has also been implicated in other biological processes, however its specific function and molecular mechanisms of action remain to be elucidated. The interaction between CD81 and its binding partner proteins may underlie the CD81 involvement in a variety of cellular processes and modulate CD81/interactors specific functions. Interestingly, in a Yeast two Hybrid system previously performed in our lab, CD81 has emerged as a putative interactor of the Amyloid Precursor Protein (APP). In the work here described, bioinformatics analyses of CD81 interacting proteins were performed and the retrieved information used to construct a protein-protein interaction network, as well as to perform Gene Ontology enrichment analyses. CD81 expression was further evaluated in CHO, GC-1 and SH-SY5Y cell lines, and in human sperm cells. Additionally, its subcellular localization was analyzed in sperm cells and in the neuronal-like SH-SY5Y cell line. Subsequently, coimmunoprecipitation assays were performed in CHO and SH-SY5Y cells to attempt to prove the physical interaction between CD81 and APP. A functional interaction between these two proteins was accessed thought the analyses of the effects of CD81 overexpression on APP levels. A co-localization analysis of CD81 and some interactors proteins retrieved from the bioinformatics analyses, such as APP, AKT1 and cytoskeleton-related proteins, was also performed in sperm cells and in SH-SY5Y cells. The effects of CD81 in cytoskeleton remodeling was evaluated in SH-SY5Y cells through monitoring the effects of CD81 overexpression in actin and tubulin levels, and analyzing the colocalization between overexpressed CD81 and F-actin. Our results showed that CD81 is expressed in all cell lines tested, and also provided the first evidence of the presence of CD81 in human sperm cells. CD81 immunoreactivity was predominantly detected in the sperm head, including the acrosome membrane, and in the midpiece, where it co-localized with APP, as well as in the post-acrosomal region. Furthermore, CD81 co-localizes with APP in the plasma membrane and in cellular projections in SH-SY5Y cells, where CD81 overexpression has an influence on APP levels, also visible in CHO cells. The analysis of CD81 interacting proteins such as AKT1 and cytoskeletonrelated proteins showed that CD81 is involved in a variety of pathways that may underlie cytoskeleton remodeling events, related to processes such as sperm motility, cell migration and neuritogenesis. These results deepen our understanding on the functions of CD81 and some of its interactors in sperm and neuronal cells.

FTIR, a potential tool to dementia diagnosis trough analysis of plasma

Nowadays it is still difficult to perform an early and accurate diagnosis of dementia, therefore many research focus on the finding of new dementia biomarkers that can aid in that purpose. So scientists try to find a noninvasive, rapid, and relatively inexpensive procedures for early diagnosis purpose. Several studies demonstrated that the utilization of spectroscopic techniques, such as Fourier Transform Infrared Spectroscopy (FTIR) and Raman spectroscopy could be an useful and accurate procedure to diagnose dementia. As several biochemical mechanisms related to neurodegeneration and dementia can lead to changes in plasma components and others peripheral body fluids, blood-based samples and spectroscopic analyses can be used as a more simple and less invasive technique. This work is intended to confirm some of the hypotheses of previous studies in which FTIR was used in the study of plasma samples of possible patient with AD and respective controls and verify the reproducibility of this spectroscopic technique in the analysis of such samples. Through the spectroscopic analysis combined with multivariate analysis it is possible to discriminate controls and demented samples and identify key spectroscopic differences between these two groups of samples which allows the identification of metabolites altered in this disease. It can be concluded that there are three spectral regions, 3500-2700 cm -1, 1800-1400 cm-1 and 1200-900 cm-1 where it can be extracted relevant spectroscopic information. In the first region, the main conclusion that is possible to take is that there is an unbalance between the content of saturated and unsaturated lipids. In the 1800-1400 cm-1 region it is possible to see the presence of protein aggregates and the change in protein conformation for highly stable parallel β-sheet. The last region showed the presence of products of lipid peroxidation related to impairment of membranes, and nucleic acids oxidative damage. FTIR technique and the information gathered in this work can be used in the construction of classification models that may be used for the diagnosis of cognitive dysfunction.