Reabilitação de pontes rodoviárias de betão armado

Cada vez mais na atualidade a reabilitação de obras de arte é de grande importância. Muitas dessas estruturas apresentam degradações e problemas estruturais consideráveis, no qual necessitam urgentemente de ações de reparação e de reforço para garantir um nível de segurança. Em Portugal só se deu uma maior importância à conservação e a reabilitação em obras de arte após a um colapso de uma ponte no concelho de Castelo de Paiva que levou à perda de vidas humanas. Este trabalho preocupa-se a dar a conhecer os vários tipos de patologias que poderão ocorrer na vida de uma ponte, desde da fase conceção e projeto até a fase de serviço, com o intuito de poder sensibilizar para futuras construções de pontes. De forma esquemática apresentam-se os diferentes tipos de inspeções, ensaios e monitorização, cujo através destes, consegue-se acompanhar o estado de conservação e o aparecimento de eventuais anomalias, que possam implicar possíveis restrições ao tráfego ou até mesmo o encerramento. De modo a poder garantir as condições de segurança e durabilidade das pontes de betão armado, ilustram-se algumas soluções e técnicas de reparação e de reforço, salientando que estas soluções e técnicas dependem da natureza e grau das anomalias, da necessidade ou não de manter a circulação de veículos e de fatores técnicos e económicos. De modo a poder exemplificar alguns aspetos importantes, nomeadamente, conservação, patologias, reparação e reforço estrutural, abordam-se duas obras de arte de Portugal Continental (um viaduto e uma ponte) que foram sujeitas a intervenções devido ao seu estado de degradação e a problemas estruturais. Por fim, apresentam-se algumas obras de arte pertencentes à Região Autónoma da Madeira (RAM) onde foram detetadas alguns problemas. Neste sentido, expõem-se algumas soluções às patologias abordadas no contacto visual, de forma a expor o conhecimento adquirido ao longo deste trabalho.

Os efeitos do plasma não térmico à pressão atmosférica no desenvolvimento larval e anomalias morfológicas de Drosophila Melanogaster

O plasma não térmico nos cuidados de saúde é um campo emergente que tem as suas raízes na ciência de plasmas. Este tipo de investigação tem crescido rapidamente e é agora objeto de um amplo esforço de pesquisa interdisciplinar envolvendo a medicina, a biologia, a física, a química e a engenharia. Têm sido feitos vários trabalhos de modo a elucidar quais as interações das espécies produzidas pelo plasma com os sistemas vivos. É evidente que o mecanismo da interação do plasma com os sistemas vivos é complexo, em parte devido à complexidade do plasma mas principalmente devido à enorme complexidade da biologia. O principal objetivo desta dissertação foi observar os efeitos do plasma não térmico à pressão atmosférica (PNTPAs) no desenvolvimento larval e anomalias morfológicas de Drosophila melanogaster. Para o efeito, foram expostas e analisadas fenotipicamente 2.566 larvas após exposição, dos diferentes estádios (1.º, 2.º e 3.º) de desenvolvimento. Os testes foram realizados com aplicações de plasma com e sem ultra violeta, em duas linhas diferentes de Drosophila; uma linha selvagem preparada por nós e uma linha laboratorial. A análise fenotípica revelou que após exposição as larvas apresentavam alterações no fenótipo e no comportamento que não foram observadas no controlo, nomeadamente anomalias nas mudas, traqueias partidas, formação de massas melanóticas que podiam persistir até à fase adulta, excesso de gotículas lipídicas, atraso no desenvolvimento, comportamento de não alimentação e formação de pupa imatura que levava à formação de pupa precoce e morte pupal. Na fase pupal, as anomalias mais comuns estavam relacionadas com a forma do pupário (causadas pela pipação prematura), apresentando um desenvolvimento aberrante. Entre os vários fenótipos observados, o mais significativo foi o criptocefálico (alterações na eversão dos discos imaginais) levando à morte pupal. Nos adultos, as principais anomalias morfológicas foram registadas na formação e segmentação das patas, na forma e padrão das nervuras das asas e na formação do tórax. A similaridade destes resultados com trabalhos publicados relacionados com a hormona esteróide ecdisona indicam que provavelmente o PNTPA poderá ter influenciado a biossíntese e/ou a regulação da ecdisona, a principal hormona que regula o desenvolvimento e a metamorfose em Drosophila.

Papel da proteína de reparo XPC na regulação das proteínas de reparo APE1, OGG1 e PARP-1 em células humanas e de camundongos

studies using UV as a source of DNA damage. However, even though unrepaired UV-induced DNA damages are related to mutagenesis, cell death and tumorigenesis, they do not explain phenotypes such as neurodegeneration and internal tumors observed in patients with syndromes like Xeroderma Pigmentosum (XP) and Cockayne Syndrome (CS) that are associated with NER deficiency. Recent evidences point to a role of NER in the repair of 8-oxodG, a typical substrate of Base Excision Repair (BER). Since deficiencies in BER result in genomic instability, neurodegenerative diseases and cancer, it was investigated in this research the impact of XPC deficiency on BER functions in human cells. It was analyzed both the expression and the cellular localization of APE1, OGG1 e PARP-1, the mainly BER enzymes, in different NER-deficient human fibroblasts. The endogenous levels of these enzymes are reduced in XPC deficient cells. Surprisingly, XP-C fibroblasts were more resistant to oxidative agents than the other NER deficient fibroblasts, despite presenting the highest of 8-oxodG. Furthermore, subtle changes in the nuclear and mitochondrial localization of APE1 were detected in XP-C fibroblasts. To confirm the impact of XPC deficiency in the regulation of APE1 and OGG1 expression and activity, we constructed a XPC-complemented cell line. Although the XPC complementation was only partial, we found that XPC-complemented cells presented increased levels of OGG1 than XPC-deficient cells. The extracts from XPC-complemented cells also presented an elevated OGG1 enzimatic activity. However, it was not observed changes in APE1 expression and activity in the XPCcomplemented cells. In addition, we found that full-length APE1 (37 kDa) and OGG1- α are in the mitochondria of XPC-deficient fibroblasts and XPC-complemented fibroblasts before and after induction of oxidative stress. On the other hand, the expression of APE1 and PARP-1 are not altered in brain and liver of XPC knockout mice. However, XPC deficiency changed the APE1 localization in hypoccampus and hypothalamus. We also observed a physical interaction between XPC and APE1 proteins in human cells. In conclusion, the data suggest that XPC protein has a role in the regulation of OGG1 expression and activity in human cells and is involved mainly in the regulation of APE1 localization in mice. Aditionally, the response of NER deficient cells under oxidative stress may not be only associated to the NER deficiency per se, but it may include the new functions of NER enzymes in regulation of expression and cell localization of BER proteins

Análise da estabilidade genética de células-tronco mesenquimais humanas

Human multipotent mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have become an important and attractive therapeutic tool since they are easily isolated and cultured, have in vitro expansion potential, substantial plasticity and secrete bioactive molecules that exert trophic effects. The human umbilical cord as a cell source for cell therapy will help to avoid several ethical, political, religious and technical issues. One of the main issues with SC lines from different sources, mainly those of embryonic origin, is the possibility of chromosomal alterations and genomic instability during in vitro expansion. Cells isolated from one umbilical cord exhibited a rare balanced paracentric inversion, likely a cytogenetic constitutional alteration, karyotype: 46,XY,inv(3)(p13p25~26). Important genes related to cancer predisposition and others involved in DNA repair are located in 3p25~26. Titanium is an excellent biomaterial for bone-implant integration; however, the use can result in the generation of particulate debris that can accumulate in the tissues adjacent to the prosthesis, in the local bone marrow, in the lymph nodes, liver and spleen. Subsequently may elicit important biological responses that aren´t well studied. In this work, we have studied the genetic stability of MSC isolated from the umbilical cord vein during in vitro expansion, after the cryopreservation, and under different concentrations and time of exposition to titanium microparticles. Cells were isolated, in vitro expanded, demonstrated capacity for osteogenic, adipogenic and chondrogenic differentiation and were evaluated using flow cytometry, so they met the minimum requirements for characterization as MSCs. The cells were expanded under different concentrations and time of exposition to titanium microparticles. The genetic stability of MSCs was assessed by cytogenetic analysis, fluorescence in situ hybridization (FISH) and analysis of micronucleus and other nuclear alterations (CBMN). The cells were able to internalize the titanium microparticles, but MSCs preserve their morphology, differentiation capacity and surface marker expression profiles. Furthermore, there was an increase in the genomic instability after long time of in vitro expansion, and this instability was greater when cells were exposed to high doses of titanium microparticles that induced oxidative stress. It is necessary always assess the risks/ benefits of using titanium in tissue therapy involving MSCs, considering the biosafety of the use of bone regeneration using titanium and MSCs. Even without using titanium, it is important that the therapeutic use of such cells is based on analyzes that ensure quality, security and cellular stability, with the standardization of quality control programs appropriate. In conclusion, it is suggested that cytogenetic analysis, FISH analysis and the micronucleus and other nuclear alterations are carried out in CTMH before implanting in a patient

Transcriptoma diferencial entre células-tronco mesenquimais humanas jovens e senescentes

Human mesenchymal stem cells (MSC) are powerful sources for cell therapy in regenerative medicine. The long time cultivation can result in replicative senescence or can be related to the emergence of chromosomal alterations responsible for the acquisition of tumorigenesis features in vitro. In this study, for the first time, the expression profile of MSC with a paracentric chromosomal inversion (MSC/inv) was compared to normal karyotype (MSC/n) in early and late passages. Furthermore, we compared the transcriptome of each MSC in early passages with late passages. MSC used in this study were obtained from the umbilical vein of three donors, two MSC/n and one MSC/inv. After their cryopreservation, they have been expanded in vitro until reached senescence. Total RNA was extracted using the RNeasy mini kit (Qiagen) and marked with the GeneChip ® 3 IVT Express Kit (Affymetrix Inc.). Subsequently, the fragmented aRNA was hybridized on the microarranjo Affymetrix Human Genome U133 Plus 2.0 arrays (Affymetrix Inc.). The statistical analysis of differential gene expression was performed between groups MSC by the Partek Genomic Suite software, version 6.4 (Partek Inc.). Was considered statistically significant differences in expression to p-value Bonferroni correction ˂.01. Only signals with fold change ˃ 3.0 were included in the list of differentially expressed. Differences in gene expression data obtained from microarrays were confirmed by Real Time RT-PCR. For the interpretation of biological expression data were used: IPA (Ingenuity Systems) for analysis enrichment functions, the STRING 9.0 for construction of network interactions; Cytoscape 2.8 to the network visualization and analysis bottlenecks with the aid of the GraphPad Prism 5.0 software. BiNGO Cytoscape pluggin was used to access overrepresentation of Gene Ontology categories in Biological Networks. The comparison between senescent and young at each group of MSC has shown that there is a difference in the expression parttern, being higher in the senescent MSC/inv group. The results also showed difference in expression profiles between the MSC/inv versus MSC/n, being greater when they are senescent. New networks were identified for genes related to the response of two of MSC over cultivation time. Were also identified genes that can coordinate functional categories over represented at networks, such as CXCL12, SFRP1, xvi EGF, SPP1, MMP1 e THBS1. The biological interpretation of these data suggests that the population of MSC/inv has different constitutional characteristics, related to their potential for differentiation, proliferation and response to stimuli, responsible for a distinct process of replicative senescence in MSC/inv compared to MSC/n. The genes identified in this study are candidates for biomarkers of cellular senescence in MSC, but their functional relevance in this process should be evaluated in additional in vitro and/or in vivo assays

Proteômica comparativa de linhagens celulares humanas expostas a estresse oxidativo induzido por riboflavina fotossensibilizada

Reactive oxygen species (ROS) are continuously generated and can be derived from cellular metabolism or induced by exogenous factors, in addition, have the capacity to damage molecules like DNA and proteins. BER is considered the main route of DNA damage oxidative repair, however, several studies have demonstrated the importance of the proteins participation of other ways to correct these injuries. NER enzymes deficiency, such as CSB and XPC, acting in the damage recognition step in the two subways this system influences the effectiveness of oxidative damage repair. However, the mechanisms by which cells deficient in these enzymes respond to oxidative stress and its consequences still need to be better understood. Thus, the aim of this study was to perform a proteomic analysis of cell lines proficient and deficient in NER, exposed to oxidative stress, in order to identify proteins involved, directly or not, in response to oxidative stress and DNA repair. For this, three strains of human fibroblasts, MRC5-SV, CS1AN (CSBdeficient) and XP4PA (XPC-deficient) were treated with photosensitized riboflavin and then carried out the differentially expressed proteins identification by mass spectrometry. From the results, it was observed in MRC5-SV increase expression in most of the proteins involved in cellular defense, an expected response to a normal cell line subjected to stress. CS1AN showed a response disjointed, it is not possible to establish many interactions between the proteins identified, may be one explanation for their sensitivity to treatment with riboflavin and other oxidants and increased cell death probably by induction of pro-apoptotic pathways. Already XP4PA showed higher expression of apoptosis-blocking proteins, as there was inhibition or reduced expression of others involved with the activation of this process, suggesting the activation of an anti-apoptotic mechanism in this lineage, which may help explain the high susceptibility to develop cancers in XPC individuals. These results also contribute to elucidate action mechanisms of NER in oxidative damage and the understanding of important routes in the oxidative stress correlation, repair and malignant tumors formation

Clara Nunes como obra de arte: a epistemologia das ciências humanas a partir da cultura musical

The thesis has its largest array in the reorganization of science that is built from the middle of the last century and its horizon reconnection between scientific culture and humanistic culture, and the dialogue between science, art and literature. This epistemological regeneration view of the scientific paradigm incorporates the poetic language and sociological analysis, and brings out a complex, open and transdisciplinary narrative. To undertake this exercise as interlocutors we have thinkers like Nietzsche, Lévi-Strauss, Edgar Morin and Bruno Latour, to name a few, and as a reference for analyzing the entire artistic production of one of the icons of Brazilian music, Clara Nunes. It is problematized up in this work, through the singer s discography, lyrics and fragments of her biography, the construction of a social character that politicized culture, increased the mestizo consciousness of popular imagery, and exceeded the excessively prosaic narratives of the academic and scientific culture. The central argument of the thesis recognizes a Hybrid Subject Clara Nunes, as indeed is what is expected of the politically engaged intellectual in the 21st century

Uso e conservação da fauna por populações humanas no Rio Grande do Norte, Nordeste do Brasil

Muitos países subdesenvolvidos são caracterizados por uma significativa diversidade sociocultural e por uma acentuada pobreza em grande parte da população, que fazem uso da caça de animais como uma fonte alternativa de alimento e para outros fins. O estudo objetivou investigar se a distância das populações humanas da zona rural em relação à zona urbana e fatores socioeconômicos podem ser preditores de um maior conhecimento sobre as espécies da fauna utilizadas. Além disso, o estudo objetivou também testar uma ferramenta capaz de gerar uma lista da fauna prioritária para a conservação local a partir do conhecimento local. Os dados foram coletados por meio de 30 entrevistas utilizando formulários semi-estruturados em quatro comunidades rurais no município de Pedro Avelino, RN. Foram calculados o fator de consenso dos informantes (ICF), a importância relativa (IR) e o nível de fidelidade (FL). Adicionalmente, foi calculado o índice de Shannon-Wiener para as categorias de uso: estimação, alimentar, medicinal e para a multiplicidade de uso por informante. Os entrevistados reconhecem 83 espécies pertencentes a 45 famílias de animais. As espécies citadas pertencem a seis grandes grupos taxonômicos: aves, mamíferos, répteis, anfíbios, peixes e insetos. O conhecimento sobre a utilização de animais para as categorias de uso definidas neste trabalho não mostrou relação significativa com os fatores socioeconômicos e nem com a distância das comunidades da zona rural para a zona urbana de Pedro Avelino. Os resultados sugerem, portanto, que a pobreza nivela o conhecimento sobre o uso da fauna independente da distância que as comunidades encontram-se da zona urbana. As listas geradas permitirão direcionar estudos ecológicos mais aprofundados visando à conservação das espécies do bioma Caatinga. Desta forma, é importante considerar as interações existentes entre as pessoas e os animais para melhor entender a dinâmica de utilização e fornecer, assim, subsídios para a conservação das espécies.

Uso da biodiversidade por populações humanas na caatinga e na Mata Atlântica, Rio Grande do Norte, Nordeste do Brasil

The use of animals is a common activity but it can be really variable between human populations. This work analyzed the knowledge about useful animal species between human communities in Caatinga and Atlantic Forest considering the influence of socioeconomics and environmental factors on the species selection. Forty people were interviewed in Caraúbas community (Atlantic Forest) and thirty were interviewed in Barrocas community (Caatinga). A total of 193 animals were cited. The communities showed a low degree of similarity (20%) and the general diversity of mentioned species was significantly bigger in Caraúbas. Men cited a diversity of species greater than women for food category in Caraúbas and pet category in Caraúbas and Barrocas. Women cited greater species diversity for medicinal use in Caraúbas. In general, in both communities, the age group >60 years showed a greater knowledge than the others. The multiple regression analysis showed the age as the only socioeconomic factor which influenced significantly the knowledge about animals for the food category in both communities. Considering the richness of known species, there were differences only between gender for sum of uses and food category in Caraúbas and for the pet category in Barrocas with men showing a greater knowledge than women. The results showed that local environmental differences are fundamental at the species selection but they also showed that within communities there are other factors like gender and age that tend to influence the knowledge about species. It is important to consider cultural, economic, social and environmental factors to understand better the interactions between people and animals so that this information can be used to facilitate the implementation of conservation measures

Sistema de supervisão aérea baseado em navegação visual para detecção de anomalias em instalações de petróleo e gás

This work deals with the development of a prototype of a helicopter quadrotor for monitoring applications in oil facilities. Anomaly detection problems can be resolved through monitoringmissions performed by a suitably instrumented quadrotor, i.e. infrared thermosensors should be embedded. The proposed monitoring system aims to reduce accidents as well as to make possible the use of non-destructive techniques for detection and location of leaks caused by corrosion. To this end, the implementation of a prototype, its stabilization and a navigation strategy have been proposed. The control strategy is based on dividing the problem into two control hierarchical levels: the lower level stabilizes the angles and the altitude of the vehicle at the desired values, while the higher one provide appropriate references signals to the lower level in order the quadrotor performs the desired movements. The navigation strategy for helicopter quadrotor is made using information provided by a acquisition image system (monocular camera) embedded onto the helicopter. Considering that the low-level control has been solved, the proposed vision-based navigation technique treats the problem as high level control strategies, such as, relative position control, trajectory generation and trajectory tracking. For the position control we use a control technique for visual servoing based on image features. The trajectory generation is done in a offline step, which is a visual trajectory composed of a sequence of images. For the trajectory tracking problem is proposed a control strategy by continuous servovision, thus enabling a navigation strategy without metric maps. Simulation and experimental results are presented to validate the proposal

Técnicas neuronais de agrupamentos aplicadas à detecção de anomalias em mamogramas digitais

This work proposes the development of an intelligent system for analysis of digital mammograms, capable to detect and to classify masses and microcalcifications. The digital mammograms will be pre-processed through techniques of digital processing of images with the purpose of adapting the image to the detection system and automatic classification of the existent calcifications in the suckles. The model adopted for the detection and classification of the mammograms uses the neural network of Kohonen by the algorithm Self Organization Map - SOM. The algorithm of Vector quantization, Kmeans it is also used with the same purpose of the SOM. An analysis of the performance of the two algorithms in the automatic classification of digital mammograms is developed. The developed system will aid the radiologist in the diagnosis and accompaniment of the development of abnormalities