Novos recursos tecnológicos aplicados às redes de abastecimento de água e de drenagem de águas residuais: tecnologias sem abertura de vala

A presente dissertação tem como objectivo a pesquisa sobre as tecnologias actuais e em desenvolvimento de detecção, inspecção e reabilitação de tubagens existentes e de instalação de novas tubagens, sem recorrer à abertura de vala. Na perspectiva do dono de obra, esta representa uma ferramenta para auxílio na ponderação e análise de métodos propostos e na tomada de decisão da sua adequação quer em termos técnicos e funcionais quer em termos económicos. Esta serve também para auxiliar a actividade de fiscalização de obras nomeadamente na inspecção, reabilitação e instalação de tubagens. Os temas, alvo de pesquisa são os quatro capítulos principais que compõem esta dissertação. O primeiro capítulo incide numa pesquisa sobre os métodos desenvolvidos para a inspecção do estado de conservação dos diversos tipos de tubagens enterradas. A pesquisa avançou conforme surgia a necessidade de alcançar novas tecnologias que permitissem transpor os novos obstáculos encontrados. O segundo capítulo aborda a complexidade da reabilitação de tubagens enterradas e o desenvolvimento das diversas técnicas que têm vindo a aumentar o "leque" de possibilidades e a facilitar a resolução dos diversos problemas. Ao analisar cada situação o método escolhido tem de assentar em quatro bases principais e manter o equilíbrio entre elas: a eficácia, o custo, a rapidez e o impacto à superfície. A detecção e localização de tubagens enterradas, tema desenvolvido no terceiro capítulo, é uma ferramenta fundamental na eliminação de problemas e custos desnecessários na fase de execução da obra. O seu domínio é fundamental e a sua problemática está na existência de interferências que prejudicam o desempenho dos equipamentos. O último capítulo descreve as principais tecnologias de perfuração para instalação de novas tubagens atendendo ao tipo de solo envolvente, diâmetro, material, extensão da tubagem a instalar ou do obstáculo a transpor e o impacto provocado à superfície

Differential HFE Gene Expression is Regulated by Alternative Splicing in Human Tissues

Background - The pathophysiology of HFE-derived Hereditary Hemochromatosis and the function of HFE protein in iron homeostasis remain uncertain. Also, the role of alternative splicing in HFE gene expression regulation and the possible function of the corresponding protein isoforms are still unknown. The aim of this study was to gain insights into the physiological significance of these alternative HFE variants. Methodology/Principal Findings - Alternatively spliced HFE transcripts in diverse human tissues were identified by RT-PCR, cloning and sequencing. Total HFE transcripts, as well as two alternative splicing transcripts were quantified using a real-time PCR methodology. Intracellular localization, trafficking and protein association of GFP-tagged HFE protein variants were analysed in transiently transfected HepG2 cells by immunoprecipitation and immunofluorescence assays. Alternatively spliced HFE transcripts present both level- and tissue-specificity. Concerning the exon 2 skipping and intron 4 inclusion transcripts, the liver presents the lowest relative level, while duodenum presents one of the highest amounts. The protein resulting from exon 2 skipping transcript is unable to associate with β2M and TfR1 and reveals an ER retention. Conversely, the intron 4 inclusion transcript gives rise to a truncated, soluble protein (sHFE) that is mostly secreted by cells to the medium in association with β2M. Conclusions/Significance - HFE gene post-transcriptional regulation is clearly affected by a tissue-dependent alternative splicing mechanism. Among the corresponding proteins, a sHFE isoform stands out, which upon being secreted into the bloodstream, may act in remote tissues. It could be either an agonist or antagonist of the full length HFE, through hepcidin expression regulation in the liver or by controlling dietary iron absorption in the duodenum.This work was partially supported by Fundação para a Ciência e a Tecnologia (FCT; grant PTDC/SAU/GMG/64494/2006) and Programa de Financiamento Plurianual do CIGMH. RM and BS were supported by FCT fellowships (SFRH/BD/21340/2005 and SFRH/BD/60718/2009)

A soluble HFE splice variant seems to regulate the expression of duodenal cytochrome b and hephaestin contributing to iron metabolism regulation

INTRODUCTION: Hereditary Hemochromatosis is an autosomal recessive disorder characterized by excessive intestinal iron absorption and pathological iron deposition in organs such as liver, heart and pancreas. The disease is predominantly caused by homozygosity for the p.C282Y mutation in HFE, which impairs protein association with its chaperone beta-2 microglobulin (B2M), for correct folding and traffic to the cell surface. Several alternative HFE transcripts have been reported but their functional significance remains elusive. Since we have identified an alternative HFE transcript due to intron 4 inclusion, we aimed to investigate its physiological role on iron homeostasis. MATERIALS AND METHODS: We analysed the expression level of the alternative transcript in several human tissues by quantitative Real-Time PCR. In addition, we produced the corresponding GFP-tagged HFE protein variant. HepG2 cells were transfected with this construct and protein cellular location analyzed by immunofluorescence using several antibodies. In parallel, immunoprecipitation was performed. Finally, the variant was over-expressed in a duodenal cell line (Hutu-80) under normal and iron overload conditions and the expression of several iron metabolism related genes (TFR1, DMT1, CYBRD1, SLC40A1 and HEPH) was evaluated by quantitative Real-Time PCR. RESULTS: We have found that the HFE_intron 4 inclusion transcript has an ubiquitous expression, being its relative expression higher in duodenum and lower in the liver. Also, we found that it gives rise to a truncated soluble protein (sHFE) that is secreted by cells maintaining its interaction with B2M. Its overexpression in HuTu-80 cells revealed that the sHFE is able to down-regulate the duodenal cytochrome b (CYBRD1) expression, as it happens with the HFE_full length protein. Also, it seems to play a specific role in the regulation of hephaestin (HEPH) expression. CONCLUSIONS: Through this study we might have unveiled the contribution of the sHFE variant to iron homeostasis. In fact, sHFE may be secreted into the bloodstream and act in remote tissues such as duodenum, down-regulating the expression of some of the iron metabolism related genes, as CYBRD1 and HEPH, and consequently reducing dietary iron absorption, preventing iron overload and contributing to iron metabolism regulation. Partially funded by FCT: PTDC/SAU-GMG/64494/2006; CIGMH; SFRH/BD/21340/2005 and SFRH/BD/60718/2009. The authors declare no competing interests

Duodenal Cytochrome B and Hephaestin Expression is Regulated by the Soluble HFE Isoform

INTRODUCTION: Hereditary Hemochromatosis is an autosomal recessive disorder characterized by excessive intestinal iron absorption and iron deposition in organs such as liver, heart and pancreas, potentially leading to cirrhosis, hepatocelular carcinoma, diabetes, cardiac failure and arthritis. This disorder is mainly due to mutations in HFE gene. HFE protein associates with beta-2 microglobulin (B2M) for trafficking to the cell surface. However, the HFE’s role on iron homeostasis is not completely cleared. It may regulate hepcidin expression in the liver and iron trafficking in the duodenum. Several HFE alternative splicing transcripts have been reported, but their structural and functional characterization have been poorly studied. MATERIALS AND METHODS: Aiming to investigate the putative biological role of an alternative HFE transcript originated by the intron 4 inclusion, we measured its expression level in several human tissues by quantitative Real-Time PCR. Also, we produced the corresponding GFP-tagged HFE variant. HepG2 cells were transfected with this construct and protein cellular location analyzed by immunofluorescence, using B2M, TfR1 and calnexin antibodies. In parallel, immunoprecipitation was performed. Finally the intron 4 inclusion variant was over-expressed in a human duodenum adenocarcinoma cell line (Hutu-80) under normal and iron overload conditions and the expression of several iron metabolism genes (TFR1, DMT1, DCYTB, SLC40A1 and HEPH) evaluated by quantitative Real-Time PCR. RESULTS: We have found that the intron 4 inclusion transcript has an ubiquitous expression in the analyzed tissues, being its relative expression higher in duodenum and lower in the liver. Also, we found that this variant gives rise to a truncated protein (sHFE) that is secreted by the cells and is able to maintain its interaction with B2M. Its overexpression in HuTu-80 cells showed that sHFE down-regulates the duodenal cytochrome b (CYBRD1) expression in about 20% independently of cellular iron status, as it happens with the HFE_full length protein. Also, sHFE seems to be involved in the down-regulation of hephaestin (HEPH) expression, being its effect higher in the presence of iron overload (reduction of ~40 and ~50%, respectively). CONCLUSIONS: Through this study we might have unveiled the contribution of the HFE’s intron 4 inclusion splice variant to the maintenance of iron homeostasis. sHFE may be secreted into the bloodstream and act in remote tissues such as the duodenum, down-regulating the expression of some of the iron metabolism related genes, as CYBRD1 and HEPH, and consequently reducing dietary iron absorption. Also we are currently exploring the hypothesis of a possible effect of sHFE in the expression of other iron metabolism related genes in hepatic cells and macrophages.Partially funded by FCT: PTDC/SAU-GMG/64494/2006; Pest-OE/SAU/UI0009/2011; SFRH/BD/21340/2005 and SFRH/BD/60718/200

Adequação dos exames de radiologia solicitados por um departamento de emergência: um estudo retrospetivo

Imaging tests are essential for diagnosis in the emergency context and convey clinical information that is essential to assess the appropriateness of the tests and improve their interpretation. Therefore, we aimed to analyze the imaging tests requested by the Emergency Department in a district hospital.info:eu-repo/semantics/publishedVersio

Temperature responsiveness of gilthead sea bream bone; an in vitro and in vivo approach

This study aimed to characterize the molecules involved in osteogenesis in seabream and establish using in vitro/in vivo approaches the responsiveness of selected key genes to temperature. The impact of a temperature drop from 23 to 13 degrees C was evaluated in juvenile fish thermally imprinted during embryogenesis. Both, in vitro/in vivo, Fib1a, appeared important in the first stages of bone formation, and Col1A1, ON and OP, in regulating matrix production and mineralization. OCN mRNA levels were up-regulated in the final larval stages when mineralization was more intense. Moreover, temperature-dependent differential gene expression was observed, with lower transcript levels in the larvae at 18 degrees C relative to those at 22 degrees C, suggesting bone formation was enhanced in the latter group. Results revealed that thermal imprinting affected the long-term regulation of osteogenesis. Specifically, juveniles under the low and low-to-high-temperature regimes had reduced levels of OCN when challenged, indicative of impaired bone development. In contrast, gene expression in fish from the high and high-to-low-temperature treatments was unchanged, suggesting imprinting may have a protective effect. Overall, the present study revealed that thermal imprinting modulates bone development in seabream larvae, and demonstrated the utility of the in vitro MSC culture as a reliable tool to investigate fish osteogenesis."Ministerio de Economia y Competitividad" (MINECO) [BES-2015-074654]; Portuguese Science Foundation (FCT) [SFRH/BPD/111512/2015, SFRH/BD/81625/2011]; MINECO, Spain [AGL2010-17324, AGL2014-57974-R]; "Generalitat de Catalunya" (XRAq); Generalitat de Catalunya [2014SGR-01371]; FCT, Portugal [CCMAR/Multi/04326/2013]; European Union [LIFECYCLE EU-FP7 222719]info:eu-repo/semantics/publishedVersio

Depression and sleep quality in older adults: a meta-analysis.

The literature emphasizes depression and poor sleep quality as problems that affect many elderly individuals. However, these problems have been related in few studies and there is no meta-analysis performed so far on this relationship. The present research reviewed the studies performed on the subjective sleep quality in order to understand how it relates to depression in older adults. The review was conducted in January 2016 and comprised publications between 2005 and 2015. Based on the electronic databases Web of Science and EBSCO, we used the keywords 'sleep quality', 'depression', and 'older' to identify the empirical studies performed. After assessing the collected studies, we selected those that presented the elderly as participants, resulting in nine papers (N=3069). A random-effects method was used to evaluate the relationship between depression and sleep. We found that an older person's lack of good sleep quality is significantly related with depression. The main limitation of this study was the difficulty in collecting a greater number of studies. Future research should consider the importance of additional variables (e.g. moderators) in order to understand and investigate viable interventions for prevention and health promotion in the elderly

O ajustamento à doença crónica: aspectos conceptuais

Quando alguém é afectado por uma doença crónica tem que alterar o seu estilo de vida de modo a poder viver o melhor possível com a doença que o vai acompanhar, se não durante toda a vida, pelo menos durante grande parte da vida. Em função das características pessoais e da interacção como o meio envolvente, social e físico, alguns ajustar-se-ão melhor e mais facilmente do que outros. Estas alterações denominam-se Ajustamento ou Adaptação. Embora estes termos sejam sinónimos, por vezes a denominação expressa orientação teórica diferente: a primeira, mais interactiva, para designar o comportamento resultante dessa interacção, momento a momento, com o meio envolvente, a segunda mais estrutural. “Ajustamento” é um termo do senso comum, utilizado na linguagem de todos os dias. O dicionário diz que ajustamento e adaptação são sinónimos e, em contexto de psicologia, saúde e doenças, os dois termos também são utilizados como sinónimos. No entanto, na psicologia, por vezes, estes termos podem expressar conceitos diferentes, consoante a orientação teórica que os utiliza. O interesse pela mudança subjacente ao ajustamento ou à adaptação considera dois aspectos: a estrutura e o processo. A estrutura refere-se a factores estáveis tais como a traços de personalidade ou estrutura cognitiva e, também, a características estáveis do meio ambiente, enquanto o processo refere-se às mudanças que vão ocorrendo em consequência das interacções, momento a momento, com as situações que surgem, explicam Lazarus e Folkman (1985)

Novel galanin receptors in teleost fish: identification, expression and regulation by sex steroids

In fish, the onset of puberty, the transition from juvenile to sexually reproductive adult animals, is triggered by the activation of pituitary gonadotrophin secretion and its timing is influenced by external and internal factors that include the growth/adiposity status of the animal. Kisseptins have been implicated in the activation of puberty but peripheral signals coming from the immature gonad or associated to the metabolic/nutritional status are also thought to be involved. Additionally, there is evidence that the galaninergic system in the brain and testis of pre-pubertal male sea bass is a possible mediator involved in the translation of somatic signals leading to gonadal maturation. Here, the transcripts for four galanin receptors (GALR), named GALR1a, 1b, 2a and 2b, were isolated from European sea bass, Dicentrarchus labrax. Phylogenetic analysis confirmed the previously reported duplication of GALR1 in teleost fish, and unravelled the duplication of GALR2 in teleost fish and in some tetrapod species. Comparison with human showed that the key amino acids involved in ligand binding are present in the corresponding GALR1 and GALR2 orthologues. Transcripts for all four receptors are expressed in brain and testes of adult fish with GALR1a and GALR1b abundant in testes 15 and hardly detected in ovaries. In order to investigate whether GALR1 dimorphic expression was dependent on steroid context we evaluated the effect of 11-ketotestosterone and 17β-estradiol treatments on the receptor expression in brain and testes of pre-pubertal males. Interestingly, steroid treatments had no effect on the expression of GALRs in the brain while in the testes, GALR1a and GALR1b were significantly up regulated by 11KT. Altogether, these results support a role for the galaninergic system, in particular the GALR1 paralog in fish reproductive function

Funcionamento sexual e saúde mental em seis doenças crónicas: convergências e divergências

Ainda que a saúde mental dos doentes crónicos seja cada vez mais uma preocupação dos profissionais de saúde, não é generalizada a avaliação rotineira do seu funcionamento sexual, que poderá ter impacto sobre a sua saúde mental. O objectivo do presente estudo é explorar a relação entre funcionamento sexual e saúde mental em doentes crónicos. Foram avaliados 77 adultos com diabetes tipo 1, 40 com diabetes tipo 2, 100 com esclerose múltipla, 79 com epilepsia, 205 com obesidade e 106 com cancro, recorrendo a um Questionário Sócio-demográfico e Clínico, à Escala de Função Sexual do MSQOL-54 e à Escala de Saúde Mental do SF-36. Na amostra total, verificaram-se correlações lineares estatisticamente significativas entre Funcionamento Sexual e Saúde Mental nos dois sexos (homens: r(161)=-0,35, p<0,0001; mulheres: r(387)=-0,36, p<0,0001). Entre os homens, as correlações oscilaram entre rs(33)=-0,58 (p<0,0001) e rs(34)=-0,21 (p=0,23); entre as mulheres, entre rs(161)=-0,49 (p<0,0001) e rs(19)=-0,03 (p=0,89). Mais concretamente, nos indivíduos com diabetes tipo 1 e cancro verificaram-se correlações lineares estatisticamente significativas entre Funcionamento Sexual e Saúde Mental nos dois sexos; nos indivíduos com diabetes tipo 2 e esclerose múltipla não se verificaram correlações significativas em nenhum dos sexos; nos indivíduos com obesidade só se verificaram correlações significativas nos indivíduos do sexo feminino; nos indivíduos com epilepsia só se verificaram correlações significativas nos indivíduos do sexo masculino. Os resultados sugerem que a promoção do funcionamento sexual de doentes crónicos poderá saldar-se por uma melhoria na sua saúde mental (e vice-versa), mas não em todas as doenças crónicas analisadas.Sob o apoio da bolsa da Fundação para a Ciência e a Tecnologia PTDC/PSI/71635/2006