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Fundamental changes in the management of water resources in Portugal are now evolving. Five regional organisations termed Administracaos de Regiao Hidrographic (ARH), will be created to manage water resources within their respective geographical areas. These areas will be catchment based. As a fore-runner to the implementation of the five ARH's a foundation project has been established within the Direcao-Geral do Recursos Naturais to examine the practical implications of the new system. This project has been divided into a number of sub-projects and complementary projects to include the Tejo complementary project. The Tejo complementary project is the focus of this report. The report is to advise on the role of biology in the proposed ARH, to establish priorities for biological studies within the present Projecto de Gestao Integrada dos Recursos Hidricos da Bacia Hidrografica do Rio Tejo (PGIRH/T) and to assist with the planning of laboratory facilities for biology at the new PGIRH/T laboratory at Alges, Lisboa.

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Systems-level studies of biological systems rely on observations taken at a resolution lower than the essential unit of biology, the cell. Recent technical advances in DNA sequencing have enabled measurements of the transcriptomes in single cells excised from their environment, but it remains a daunting technical problem to reconstruct in situ gene expression patterns from sequencing data. In this thesis I develop methods for the routine, quantitative in situ measurement of gene expression using fluorescence microscopy.

The number of molecular species that can be measured simultaneously by fluorescence microscopy is limited by the pallet of spectrally distinct fluorophores. Thus, fluorescence microscopy is traditionally limited to the simultaneous measurement of only five labeled biomolecules at a time. The two methods described in this thesis, super-resolution barcoding and temporal barcoding, represent strategies for overcoming this limitation to monitor expression of many genes in a single cell. Super-resolution barcoding employs optical super-resolution microscopy (SRM) and combinatorial labeling via-smFISH (single molecule fluorescence in situ hybridization) to uniquely label individual mRNA species with distinct barcodes resolvable at nanometer resolution. This method dramatically increases the optical space in a cell, allowing a large numbers of barcodes to be visualized simultaneously. As a proof of principle this technology was used to study the S. cerevisiae calcium stress response. The second method, sequential barcoding, reads out a temporal barcode through multiple rounds of oligonucleotide hybridization to the same mRNA. The multiplexing capacity of sequential barcoding increases exponentially with the number of rounds of hybridization, allowing over a hundred genes to be profiled in only a few rounds of hybridization.

The utility of sequential barcoding was further demonstrated by adapting this method to study gene expression in mammalian tissues. Mammalian tissues suffer both from a large amount of auto-fluorescence and light scattering, making detection of smFISH probes on mRNA difficult. An amplified single molecule detection technology, smHCR (single molecule hairpin chain reaction), was developed to allow for the quantification of mRNA in tissue. This technology is demonstrated in combination with light sheet microscopy and background reducing tissue clearing technology, enabling whole-organ sequential barcoding to monitor in situ gene expression directly in intact mammalian tissue.

The methods presented in this thesis, specifically sequential barcoding and smHCR, enable multiplexed transcriptional observations in any tissue of interest. These technologies will serve as a general platform for future transcriptomic studies of complex tissues.

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Specklegram in multimode fiber has successfully been used as a sensor for detecting mechanical disturbance. Speckles in a multimode pure silica grapefruit fiber are observed and compared to that of a step-index multimode fiber, showing different features between them. The sensitivities to external disturbance of two kinds of fiber were measured, based on single-multiple-single mode (SMS) fiber structure. Experimental results show that the grapefruit fiber shows higher sensitivity than does the step-index multimode fiber. The transmission spectrum of the grapefruit fiber was measured as well, showing some oscillation features that are significantly different from that of a step-index multimode fiber. The experiments may provide suggestions to understand the mechanisms of light propagation in grapefruit fibers. (D 2008 Optical Society of America.

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Ecosystem level models are motivated by some combination of scientific and practical concerns. Those models motivated by practical concerns are likely to bear little historical relation to previous models. Mechanisms of interaction between particular species and their ecosystems vary enormously. Some species literally construct their own ecosystems. Others have more or less complex and important interactions with other species so that their presence or absence may alter the ecosystem. Prior information about the natural history of particular species can make ecosystem investigations quicker, cheaper, and more effective. The optimal resource for preparing to deal with the unlimited diversity of questions asked of ecologists would be a large' computerized databank of natural history observations for as many species as possible.

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A novel method incorporating the shielded method and the post-processing method has been proposed to fabricate the pi-phase-shilted fibre grating. Then an Er-doped pi-phase-shifted distributed feedback fibre grating laser has been fabricated using the grating. The laser threshold is 20 mW. When pumped with 90 mW light at 980 nm, the laser gives an output of 1.1 mW. Its signal-to-noise ratio is better than 60 dB. It is demonstrated that the laser is single mode operation by means of a Fabry-Perot scanning interferometer.

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Stable single-frequency and single-polarization distributed-feedback (DFB) fiber laser was realized by giving a pressure on the phase shift region of the fiber grating. The output wavelength of the DFB fiber laser is 1053 nm. When the pump power of 980 nm laser diode is 100 and 254 mW, the output power can reach 8.3 and 37.1 mW and the polarization extinction ratio was 26 and 20 dB, respectively. After chopped by Acousto-optic modulator (0.3 Hz), the pulse peak value variance is 4.65%(peak to peak) and 1.098% (RMS) for 31 min. (C) 2006 Elsevier Ltd. All rights reserved.

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Incorporating the shielded method and post-processing method, a 75 mW single frequency Yb-doped DFB fiber laser was obtained with a 250 mW laser diode pump source at 978 nm. The threshold of the laser is 2 mW. The laser is single-polarization operation and the output power fluctuation is less than 0.2 mW in one hour when the pump power is 250 mW.

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Crystallization is achieved in amorphous Ge2Sb2Te5 films upon irradiation with a single femtosecond laser pulse. Transmission electron microscopy images evidence the morphology of the crystallized spot which depends on the fluence of the ferntosecond laser pulse. Fine crystalline grains are induced at low fluence, and the coarse crystalline grains are obtained at high fluence. At the damage fluence, ablation of the films occurs. (C) 2004 Elsevier Ltd. All rights reserved.

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The morphology of materials resulting from laser irradiation of the single-layer and the multilayer amorphous Ge2Sb2Te5 films using 120 fs pulses at 800 nm was observed using scanning electron microscopy and atomic force microscopy. For the single-layer film, the center of the irradiated spot is depression and the border is protrusion, however, for the multilayer film, the center morphology changes from a depression to a protrusion as the increase of the energy. The crystallization threshold fluence of the single-layer and the multilayer film is 22 and 23 mJ/cm(2), respectively. (c) 2005 Elsevier B.V. All rights reserved.

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Esta pesquisa tem como objetivo geral, compreender as práticas desenvolvidas no cuidado realizado por enfermeiros nos Programas Nacionais de Saúde em uma unidade da rede básica de saúde no município do Rio de Janeiro. Este estudo corresponde ao desdobramento do projeto Práticas de cuidado no SUS: o papel do enfermeiro na Atenção Básica, apoiado por bolsa de produtividade em pesquisa da Universidade do Estado do Rio de Janeiro (UERJ/ Prociência). A presente dissertação trata-se de um estudo de caso realizado no âmbito da Atenção Básica, e, possui como cenário o Centro Municipal de Saúde (CMS) Milton Fontes Magarão. Os sujeitos da pesquisa corresponderam a oito enfermeiros, sendo sete entrevistas e um depoimento. Os instrumentos de coleta de dados utilizados, durante o período de julho a setembro de 2011, foram: observação sistemática, entrevista semi-estruturada, depoimento, formulário e pesquisa em fontes documentais. Para a avaliação dos resultados utilizou-se a técnica de análise temática, proposta por Minayo. No plano metodológico a análise foi qualitativa, no entanto, no tratamento dos dados, a análise aplicada se apropriou de elementos da abordagem quantitativa, com o objetivo de encontrar os núcleos de sentido contextualizados nas entrevistas. A partir dos resultados alcançados foi possível elaborar três categorias. A primeira categoria focou uma abordagem temática sobre as práticas desenvolvidas no cuidado realizado por enfermeiros nos Programas Nacionais. A segunda categoria buscou descrever a visão do enfermeiro sobre o desenvolvimento de suas práticas e do cuidado realizado, no qual, foram discutidas duas temáticas centrais: noções acerca das atividades realizadas pelos enfermeiros e noções dos enfermeiros acerca dos princípios e diretrizes para atenção à saúde. A terceira categoria direcionou-se para os fatores que interferem nas práticas e no cuidado realizado pelos enfermeiros. A prática e o cuidado realizados pelos enfermeiros foram analisados e discutidos, a partir dos relatos dos profissionais sobre suas vivências. Sendo assim, não coube a expressão de uma única forma de prática ou de uma única dimensão do cuidado, visto que essas práticas parecem estar entrelaçadas a diversos aspectos subjetivos, epistemológicos, culturais e sócio-econômicos. As práticas realizadas, independente do Programa Nacional de atuação dos enfermeiros, estão voltadas principalmente para a consulta de enfermagem, a prática educativa, a organização do fluxo de entrada, o sistema de referência e a visita domiciliar. Pôde-se inferir que diversos são os fatores que influenciam, de forma positiva ou negativa, o desenvolvimento das atividades diárias. Por fim, cabe ressaltar a necessidade de novos estudos sobre a temática a fim de estimular a construção de saberes teórico-práticos e a formulação de uma visão crítica sobre o cuidado profissional realizado pelo enfermeiro no âmbito da Atenção Básica.

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A single-longitudinal-mode (SLM) laser-diode pumped Nd: YAG laser with adjustable pulse width is developed by using the techniques of pre-lasing and changing polarization of birefingent crystal. The Q-switching voltage is triggered by the peak of the pre-lasing pulse to achieve the higher stability of output pulse energy. The output energy of more than 1 mJ is obtained with output energy stability of 3% (rms) at 100 Hz. The pulse-width can be adjusted from 30 ns to 300 ns by changing the Q-switching voltage. The probability of putting out single-longitudinal-mode pulses is almost 100%. The laser can be run over four hours continually without mode hopping.