There is a strong trend in fabricating miniaturized total analytical systems, µTAS, for various biochemical and cell biology applications. These miniaturized systems could e.g. gain better separation performances, be faster, consume less expensive reagents and be used for studies that are difficult to access in the macro world. Disposable µTAS eliminate the risk of carry-over and can be fabricated to a low cost. This work focused on the development of µTAS modules with the intentional use for miniaturized diagnostics. Modules for blood separation, desalting, enrichment, separation and ESI-MS detection were successfully fabricated. Surface coatings were additionally developed and evaluated for applications in µTAS with complex biological samples. The first heparin coating could be easily immobilized in a one-step-process, whereas the second heparin coating was aimed to form a hydrophilic surface that was able to draw blood or plasma samples into a microfluidic system by capillary forces. The last mentioned heparin surface was further utilized when developing a chip-based sensor for performing CD4-count in human blood, an important marker to determine the stage of an HIV-infection. All devices in this work were fabricated in PDMS, an elastomeric polymer with the advantage of rapid and less expensive prototyping of the microfabricated master. It was shown that PDMS could be considered as the material of choice for future commercial µTAS. The devices were intentionally produced using a low grade of fabrication complexity. It was however demonstrated that even with low complexity, it is possible to integrate several functional chip modules into a single microfluidic device.

Analysis of compounds present in complex matrices is always a challenge, which can be partly overcome by applying various sample preparation techniques prior to detection. Ideally, the extraction techniques should be as selective as possible, to minimize the concentration of interfering substances. In addition, results can be improved by efficient chromatographic separation of the sample components. The elimination of interfering substances is especially important when utilizing mass spectrometry (MS) as a detection technique since they influence the ionization yields. It is also important to optimize ionization methods in order to minimize detection limits. In the work this thesis is based upon, selective solid phase extraction (SPE) materials, a restricted access material (RAM) and graphitized carbon black (GCB) were employed for clean up and/or pre-concentration of analytes in plasma, urine and agricultural drainage water prior to liquid chromatography/mass spectrometry (LC/MS). Two SPE formats, in which GCB was incorporated in µ-traps and disks, were developed for cleaning up small and large volume samples, respectively. In addition, techniques based on use of sub-2 µm C18 particles at elevated temperatures and a linear ion trap (LIT) mass spectrometer were developed to improve the efficiency of LC separation and sensitivity of detection of 6-formylindolo[3,2-b]carbazole (FICZ) metabolites in human urine. It was also found that GCB can serve not only as a SPE sorbent, but also as a valuable surface for surface-assisted laser desorption ionization (SALDI) of small molecules. The dual functionality of GCB was utilized in a combined screening-identification/quantification procedure for fast elimination of negative samples. This may be particularly useful when processing large numbers of samples. SALDI analyses of small molecules was further investigated and improved by employing two kinds of new surfaces: oxidized GCB nanoparticles and silicon nitride.

Background: The needs of the consumers seem to develop at the same rate as the technology advancements and put more pressure on firms to produce new and innovative products at a faster pace. Research has shown that external sources can have a significant effect on the firms’ innovation performance, but the results are partial contradicting and more research is needed. Purpose: Investigate customer and supplier involvement in product innovation in Swedish manufacturing firms. Delimitations: The study was carried out to manufacturing firms in Sweden, as Swedish firms are the most innovative in Europe at this point in time. Only large and medium sized firms were under investigation as larger firms are more eager to innovate than smaller firms. Method: The research had a deductive quantitative approach. The data was collected through a questionnaire sent out by e-mail and 124 firms participated in the study. After assessing validity and reliability, the hypotheses were tested by multiple and single regression analysis. Conclusions: Firms that strives to improve their new product development process should seize their customer’s knowledge and use it to co-develop new products. Another important factor was to use lead users, as these will improve the innovativeness even more than “ordinary” customers. The study found no support for early supplier integration in the new product development process.

El propòsit d'aquesta tesi és examinar les aportacions a l'estudi de la predicció de la taxa de canvi basada en l'ús de tècniques d'aprenentatge automàtic. Aquestes aportacions es veuen facilitades i millorades per l'ús de variables econòmiques, indicadors tècnics i variables de tipus ‘business and consumer survey’. Aquesta investigació s’organitza entorn d’una recopilació de quatre articles. L'objectiu de cadascun dels quatre treballs de recerca d'aquesta tesi és el de contribuir a l'avanç del coneixement sobre els efectes i mecanismes mitjançant els quals l'ús de variables econòmiques, indicadors tècnics, variables de tipus ‘business and consumer survey’, i la selecció dels paràmetres de models predictius són capaços de millorar les prediccions de la taxa de canvi. Fent ús d'una tècnica de predicció no lineal, el primer article d'aquesta tesi es centra majoritàriament en l'impacte que tenen l'ús de variables econòmiques i la selecció dels paràmetres dels models en les prediccions de la taxa de canvi per a dos països. L'últim experiment d'aquest primer article fa ús de la taxa de canvi del període anterior i d'indicadors econòmics com a variables d'entrada en els models predictius. El segon article d'aquesta tesi analitza com la combinació de mitjanes mòbils, variables de tipus ‘business and consumer survey’ i la selecció dels paràmetres dels models milloren les prediccions del canvi per a dos països. A diferència del primer article, aquest segon treball de recerca afegeix mitjanes mòbils i variables de tipus ‘business and consumer survey’ com a variables d'entrada en els models predictius, i descarta l'ús de variables econòmiques. Un dels objectius d'aquest segon article és determinar el possible impacte de les variables de tipus ‘business and consumer survey’ en les taxes de canvi. El tercer article d'aquesta tesi té els mateixos objectius que el segon, però amb l'excepció que l'anàlisi abasta les taxes de canvi de set països. El quart article de la tesi compta amb els mateixos objectius que l'article anterior, però amb la diferència que fa ús d'un sol indicador tècnic. En general, l'enfocament d'aquesta tesi pretén examinar diferents alternatives per a millorar les prediccions del tipus de canvi a través de l'ús de màquines de suport vectorial. Una combinació de variables i la selecció dels paràmetres dels models predictius ajudaran a aconseguir aquest propòsit. / El propósito de esta tesis es examinar las aportaciones al estudio de la predicción de la tasa de cambio basada en el uso de técnicas de aprendizaje automático. Dichas aportaciones se ven facilitadas y mejoradas por el uso de variables económicas, indicadores técnicos y variables de tipo ‘business and consumer survey’. Esta investigación está organizada en un compendio de cuatro artículos. El objetivo de cada uno de los cuatro trabajos de investigación de esta tesis es el de contribuir al avance del conocimiento sobre los efectos y mecanismos mediante los cuales el uso de variables económicas, indicadores técnicos, variables de tipo ‘business and consumer survey’, y la selección de los parámetros de modelos predictivos son capaces de mejorar las predicciones de la tasa de cambio. Haciendo uso de una técnica de predicción no lineal, el primer artículo de esta tesis se centra mayoritariamente en el impacto que tienen el uso de variables económicas y la selección de los parámetros de los modelos en las predicciones de la tasa de cambio para dos países. El último experimento de este primer artículo hace uso de la tasa de cambio del periodo anterior y de indicadores económicos como variables de entrada en los modelos predictivos. El segundo artículo de esta tesis analiza cómo la combinación de medias móviles, variables de tipo ‘business and consumer survey’ y la selección de los parámetros de los modelos mejoran las predicciones del cambio para dos países. A diferencia del primer artículo, este segundo trabajo de investigación añade medias móviles y variables de tipo ‘business and consumer survey’ como variables de entrada en los modelos predictivos, y descarta el uso de variables económicas. Uno de los objetivos de este segundo artículo es determinar el posible impacto de las variables de tipo ‘business and consumer survey’ en las tasas de cambio. El tercer artículo de esta tesis tiene los mismos objetivos que el segundo, pero con la salvedad de que el análisis abarca las tasas de cambio de siete países. El cuarto artículo de esta tesis cuenta con los mismos objetivos que el artículo anterior, pero con la diferencia de que hace uso de un solo indicador técnico. En general, el enfoque de esta tesis pretende examinar diferentes alternativas para mejorar las predicciones del tipo de cambio a través del uso de máquinas de soporte vectorial. Una combinación de variables y la selección de los parámetros de los modelos predictivos ayudarán a conseguir este propósito. / The purpose of this thesis is to examine the contribution made by machine learning techniques on exchange rate forecasting. Such contributions are facilitated and enhanced by the use of fundamental economic variables, technical indicators and business and consumer survey variables as inputs in the forecasting models selected. This research has been organized in a compendium of four articles. The aim of each of these four articles is to contribute to advance our knowledge on the effects and means by which the use of fundamental economic variables, technical indicators, business and consumer surveys, and a model’s free-parameters selection is capable of improving exchange rate predictions. Through the use of a non-linear forecasting technique, one research paper examines the effect of fundamental economic variables and a model’s parameters selection on exchange rate forecasts, whereas the other three articles concentrate on the effect of technical indicators, a model’s parameters selection and business and consumer surveys variables on exchange rate forecasting. The first paper of this thesis has the objective of examining fundamental economic variables and a forecasting model’s parameters in an effort to understand the possible advantages or disadvantages these variables may bring to the exchange rate predictions in terms of forecasting performance and accuracy. The second paper of this thesis analyses how the combination of moving averages, business and consumer surveys and a forecasting model’s parameters improves exchange rate predictions. Compared to the first paper, this second paper adds moving averages and business and consumer surveys variables as inputs to the forecasting model, and disregards the use of fundamental economic variables. One of the goals of this paper is to determine the possible effects of business and consumer surveys on exchange rates. The third paper of this thesis has the same objectives as the second paper, but its analysis is expanded by taking into account the exchange rates of 7 countries. The fourth paper in this thesis takes a similar approach as the second and third papers, but makes use of a single technical indicator. In general, this thesis focuses on the improvement of exchange rate predictions through the use of support vector machines. A combination of variables and a model’s parameters selection enhances the way to achieve this purpose.

L'analyse de la composition chimique de deux orchidées de la sous-tribu des Aeridinae, Aerides rosea Lodd. ex. Lindl. & Paxton et Acampe rigida (Buch.-Ham. ex Sm.) P. F. Hunt, a été menée en ayant recours à des techniques de couplages et grâce à une stratégie de déréplication. Dix dérivés phénanthréniques ont ainsi été identifiés dans les tiges d'A. rosea, dont deux nouvellement décrits. Quatre stilbénoïdes, trois dérivés d'acides phénoliques et quatre esters d'acide cinnamique ont été identifiés dans les tiges d'A. rigida. Nous avons également effectué des dosages de traceurs dans différents échantillons de deux représentants de la tribu des Vandeae : Vanda teres (Roxb.) Lindl. et Vanda coerulea Griff. ex. Lindl., afin de mettre en évidence d'éventuelles variations de composition en fonction des facteurs environnementaux et du stade de croissance végétative. Enfin, nous avons mis au point des conditions d'isolement préparatif par chromatographie de partage centrifuge afin de purifier des marqueurs biologiques préalablement identifiés dans les tiges de Vanda teres : trois glucospyranosyloxybenzyl - malates ainsi que leur précurseur biosynthétique.

The analysis of low abundant biological molecules is often challenging due to their chemical properties, low concentration and limited sample volumes. Neuropeptides are one group of molecules that fits these criteria. Neuropeptides also play an important role in biological functions, which makes them extra interesting to analyze. A classic chemical analysis involves sampling, sample preparation, separation and detection. In this thesis, an enhanced solid supported microdialysis method was developed and used as a combined sampling- and preparation technique. In general, significantly increased extraction efficiency was obtained for all studied peptides. To be able to control the small sample volumes and to minimize the loss of neuropeptides because of unwanted adsorption onto surfaces, the subsequent analysis steps were miniaturized to a micro total analysis system (µ-TAS), which allowed sample pre-treatment, injection, separation, manipulation and detection. In order to incorporate these analysis functions to a microchip, a novel microfabrication protocol was developed. This method facilitated three-dimensional structures to be fabricated without the need of clean room facilities. The sample pre-treatment step was carried out by solid phase extraction from beads packed in the microchip. Femtomole levels of neuropeptides were detected from samples possessing the same properties as microdialysates. The developed injection system made it possible to conduct injections from a liquid chromatographic separation into a capillary electrophoresis channel, which facilitated for advanced multidimensional separations. An electrochemical sample manipulation system was also developed. In the last part, different electrospray emitter tip designs made directly from the edge of the microchip substrate were developed and evaluated. The emitters were proven to be comparable with conventional, capillary based emitters in stability, durability and dynamic flow range. Although additional developments remain, the analysis steps described in this thesis open a door to an integrated, on-line µ-TAS for neuropeptides analysis in complex biological samples.

Advances in life science require knowledge of active molecules in complex biological systems. These molecules are often only present for a certain time and at limited concentrations. Integrated micro-analytical tools for sampling, separation and mass spectrometric (MS) detection would meet these requests and are therefore continuously gaining interest. An on-line coupling of analytical functions provides shorter analysis time and less manual sample handling. In this thesis, improved compatibility of microdialysis sampling and multidimensional separations coupled to MS detection are developed and discussed. Microdialysis was used in vitro for determination of the non-protein bound fraction of the drug ropivacaine. The sampling unit was coupled on-line to capillary column liquid chromatography (LC) followed by ultraviolet or MS detection. For MS detection, the system was extended with a desalting step and an addition of internal standard. A method for MS screening of microdialysates, collected in vivo, was also developed. The method involved sampling and measurements of the chemical pattern of molecules that generally are ignored in clinical investigations. Chemometric tools were used to extract the relevant information and to compare samples from stimulated and control tissues. Complex samples often require separation in more than one dimension. On-line interfaces for sample transfer between LC and capillary electrophoresis (CE) were developed in soft poly(dimethylsiloxane) (PDMS). MS detection in the LC-CE system was optimised on frequent sampling of the CE peak or on high resolution in mass spectra using time-of-flight (TOF)MS or Fourier transform ion cyclotron resonance (FTICR)MS, respectively. Aspects on electrode positioning in the LC-CE interface led to development of an on-column CE electrode. A successful method for deactivation of the PDMS surface using a polyamine polymer was also developed. The systems were evaluated using peptides and proteins, molecules that are gaining increased attention in bioscience, and consequently also in chemical analysis.

Made available in DSpace on 2014-06-11T19:26:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-08-03Bitstream added on 2014-06-13T20:54:15Z : No. of bitstreams: 1 perinotto_rrc_me_rcla.pdf: 7856943 bytes, checksum: 17e8581630be4a2e1add4cefd13abc6f (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Derrames de óleo causam recorrentes e graves prejuízos aos ecossistemas costeiros em todo o mundo. O mapeamento de sensibilidade ambiental é componente essencial e fonte de informação primária para o planejamento de contingência, que estabelece diretrizes de ações de resposta aos derrames. O objetivo deste trabalho foi definir áreas prioritárias de proteção e zonas de sacrifício por meio do mapeamento, em escala operacional, da sensibilidade a derrames de óleo dos ambientes costeiros dos municípios de São Vicente, Santos e Guarujá (SP), baseando-se na proposta elaborada pelo Ministério do Meio Ambiente, com utilização de um Sistema de Informação Geográfica. A região apresenta elevada complexidade e relevância econômica com alto risco de ocorrência de derrames pela presença do Porto de Santos. Possui grande variedade de ecossistemas costeiros e estuarinos sensíveis que abrigam comunidades de elevada diversidade biológica. Como resultados são apresentados 3 Cartas Táticas, 21 Cartas Operacionais e Quadros Descritivos com informações de cada segmento mapeado; além do mapa e das indicações das Áreas Prioritárias e Zonas de Sacrifício, sugeridas de acordo com critérios relevantes, baseados nas características de cada segmento, e considerados a partir do conhecimento detalhado adquirido no desenvolvimento deste trabalho, que constitui importante contribuição à gestão ambiental da zona costeira estudada / Oil spills have caused recurrent and severe damages in coastal ecosystems all around the world. The environmental sensitivity mapping is an essential component and prime information source for the contingency planning, which establishes emergency response actions to oil spills. This study aimed to define high priority areas for protection and sacrifice zones by mapping, in detailed scales, the environmental sensitivity to oil spills of coastal habitats in São Vicente, Santos and Guarujá municipalities (São Paulo State). This mapping was done using a Geographic Information System and was based on the methodology and standards suggested by the Brazilian Environmental Ministry. The study area has high economic importance and complexity, with many risks of oil spills because of the presence of the “Porto de Santos”. The region shows a wide variety of sensitive estuarine and coastal ecosystems that shelter high biodiversity. As result are presented 3 ESI maps in tactical scales; 21 ESI maps in detailed scales; Descriptive Tables with information from each coastal segment mapped; and the map with indicated areas for high priority protection and sacrifice zones. These areas were suggested according to relevant criteria, based on characteristics of each segment, and considered from the detailed knowledge acquired in the development of this work that represents an important contribution to the coastal environmental management in the study area

The Oil Pollution Act of 1990 requires the development of Regional and Area Contingency Plans. For more than 20 years, the State of Florida, under both the Department of Environmental Protection and the Florida Fish and Wildlife Commission, has worked closely with the U.S. Coast Guard and the National Oceanic and Atmospheric Administration to develop these plans for coastal and marine oil spill response. Current plans, developed with local, state and federal stakeholder input, use geographic information systems (GIS) data such as location and extent of sensitive ecological, wildlife, and human-use features (termed Environmental Sensitivity Index data), pre-defined protection priorities, and spatially explicit protection strategies to support decision-making by responders (termed Geographic Response Plans). However, they are long overdue for improvements that incorporate modern oceanographic modeling techniques and integrated data from coastal ocean observing systems. Better understanding of circulation in nearshore and estuarine waters, at a scale consistent with other spatial data, is especially lacking in Area Contingency Plans. This paper identifies the gaps in readily available information on the circulation-driven causes and effects missing in current oil spill contingency planning and describes a sample methodology whereby multiple coastal and ocean spatial science disciplines are used to answer questions that no single, non-integrated discipline can answer by itself. A path forward for further integration and development of more comprehensive plans to better support coastal protection in Florida is proposed. The advances made here are applicable to other coastal regions of the world.

A Estação Ecológica (ESEC) Tupinambás, uma UC marinha de proteção integral, está localizada no litoral norte do estado de São Paulo (SP). Foi criada em 1987, sendo utilizada pela Marinha do Brasil para exercícios de tiro até recentemente no arquipélago dos Alcatrazes (São Sebastião, SP). Isto justifica a baixa quantidade de informações disponíveis sobre essa região. Através do levantamento de informações sobre a composição biológica, aspectos socioeconômicos e oceanográficos da área do arquipélago, da utilização de índices ecológicos (dominância, diversidade, riqueza e equabilidade) e multimétricos (Índice ABC e Índice de Integridade Biótica), e da elaboração da Carta de Sensibilidade Ambiental ao Derramamento de Óleo, verificou-se (1) a semelhança da composição ictiofaunística do arquipélago dos Alcatrazes em relação à Santos e São Sebastião, (2) se a região mantém a integridade biótica da ictiofauna mesmo localizando-se entre regiões impactadas, e (3) se o ponto mais sensível ecologicamente detectado pela carta SAO pode ser a área de maior suscetibilidade de ser atingida por um derramamento de óleo. Os dados analisados são provenientes de trabalhos pretéritos e coletas realizadas em setembro de 2011 e janeiro de 2014. A composição ictíica variou ao longo do tempo e períodos de coleta, respondendo à composição sedimentar e à variabilidade das propriedades hidrográficas. Foi constatada alta similaridade da ictiofauna do arquipélago com aquelas encontradas na plataforma continental de Santos e São Sebastião. Em relação à integridade biótica local, a partir da ictiofauna, as duas técnicas aplicadas refletiram o histórico no estado de conservação da ESEC, com a melhora dos índices ao longo do período estudado. A produção da carta SAO resultou em um produto único, que poderá auxiliar a gestão da UC e em planos de contenção, e em um compêndio das espécies encontradas na região, dentre as quais se encontram algumas endêmicas e muitas ameaçadas, segundo critérios nacionais e internacionais. Finalmente, espera-se que a alta biodiversidade do entorno do arquipélago, que representa uma vasta fonte de conhecimentos para o futuro, se encontre protegida pelo respeito à legislação e fiscalização da ESEC em que se encontra, pela Área Delta da Marinha do Brasil, e pela distância da costa e de fontes de contaminação. / The Tupinambás Ecological Station (TES), a Marine Protected Area (MPA) of integral protection, is located on the northern coast of São Paulo state (SP). It was established in 1987 and it has been used by the Brazilian Navy for shooting practice until recently in the Alcatrazes archipelago (São Sebastião, SP). That accounts for the low amount of the available data in this region. Using information on the biological composition, socioeconomic and oceanographic aspects variables, combined with ecological (dominance, diversity, richness and evenness) and multimetric (ABC method and Biotic Integrity Index) indexes and the preparation of the Environmental Sensitivity Index (ESI) Map, we (1) assessed the similarity of the ichthyofauna of the Alcatrazes archipelago regarding compared to Santos and São Sebastião, (2) verified whether the region retains the ichthyofauna biotic integrity despite being located between affected regions and (3) verified whether the most ecologically sensitive region detected by the ESI map is the area most likely to be hit by an oil spill. The analyzed data come from previous works and samplings carried out in September 2011 and January 2014. The ichthyofauna varied over time and sampling points. Its distribution also reflected the sedimentary composition and the variability of hydrographic properties. High similarity between the ichthyofauna of the archipelago and those found on the continental shelf off Santos and São Sebastião. Regarding the biotic integrity, the two techniques applied reflected the conservation history of the TES, with an improvement in the indexes during the study period. The production of the ESI map resulted in an unique product that can assist in the management of the MPA and containment plans, and a species compendium found in the region, among which many of them are endemic and endangered, according to national and international classifications. Finally, it is expected that the high biodiversity of the archipelago surroundings, which still represents a vast source of knowledge for the future, is protected by the Delta Area of the Brazilian Navy, by the law that ensure the TES and by the distance from the coast.