Development of a high-throughput monitoring technique of bacteria photodynamic inactivation

Summary form only given. Bacterial infections and the fight against them have been one of the major concerns of mankind since the dawn of time. During the `golden years' of antibiotic discovery, during the 1940-90s, it was thought that the war against infectious diseases had been won. However currently, due to the drug resistance increase, associated with the inefficiency of discovering new antibiotic classes, infectious diseases are again a major public health concern. A potential alternative to antibiotic treatments may be the antimicrobial photodynamic inactivation (PDI) therapy. To date no indication of antimicrobial PDI resistance development has been reported. However the PDI protocol depends on the bacteria species [1], and in some cases on the bacteria strains, for instance Staphylococcus aureus [2]. Therefore the development of PDI monitoring techniques for diverse bacteria strains is critical in pursuing further understanding of such promising alternative therapy. The present works aims to evaluate Fourier-Transformed-Infra-Red (FT-IR) spectroscopy to monitor the PDI of two model bacteria, a gram-negative (Escherichia coli) and a gram-positive (S. aureus) bacteria. For that a high-throughput FTIR spectroscopic method was implemented as generally described in Scholz et al. [3], using short incubation periods and microliter quantities of the incubation mixture containing the bacteria and the PDI-drug model the known bactericidal tetracationic porphyrin 5,10,15,20-tetrakis (4-N, N, Ntrimethylammoniumphenyl)-porphyrin p-tosylate (TTAP4+). In both bacteria models it was possible to detect, by FTIR-spectroscopy, the drugs effect on the cellular composition either directly on the spectra or on score plots of principal component analysis. Furthermore the technique enabled to infer the effect of PDI on the major cellular biomolecules and metabolic status, for example the turn-over metabolism. In summary bacteria PDI was monitored in an economic, rapid (in minutes- , high-throughput (using microplates with 96 wells) and highly sensitive mode resourcing to FTIR spectroscopy, which could serve has a technological basis for the evaluation of antimicrobial PDI therapies efficiency.

Projetos geotérmicos para usos diretos em Portugal Continental: tipologias e viabilidade económica

Os aproveitamentos geotérmicos têm vindo a aumentar significativamente em todo o mundo, sendo os Estados Unidos da América, o maior produtor desta energia proveniente do interior da Terra, com cerca de 3.187 MW de capacidade instalada. Portugal tem capacidade instalada total de 29 MW, no entanto no que se refere ao aproveitamento de “alta entalpia”, isto é, o aproveitamento geotérmico para produção elétrica, apenas se encontra no arquipélago dos Açores, na ilha de S. Miguel, onde estão instaladas e em funcionamento duas centrais geotérmicas com a potência total de 23 MW, com produção de energia de 185 GWh. Em Portugal Continental, não se consegue produzir energia elétrica devido às temperaturas existentes, restringindo esta utilização apenas ao aproveitamento de baixa entalpia (máximo de 76 ºC). Este aproveitamento normalmente é feito em cascata, segundo, predominando o aquecimento de águas sanitárias, climatização, e para termas, usando águas termominerais. Para a exploração deste recurso renovável, é necessário conhecer a hidrogeologia do país, e relacioná-la com a fracturação, e acidentes tectónicos. Portugal Continental, está divido em quatros partes distintas a nível hidrogeológico, o Maciço Antigo, a Orla Ocidental, a Bacia Tejo-Sado e a Orla Meridional. Qualquer aproveitamento geotérmico em Portugal terá de atender a estas características, potenciando também, novas explorações geotérmicas orientadas para as pessoas, respeitando os valores sociais, culturais e ambientais. Neste contexto, existem alguns complexos geotérmicos em funcionamento, outros abandonados, e muitos outros em estudo para uma breve aplicação. Um exemplo de sucesso no aproveitamento do calor geotérmico, é o complexo de Chaves, que foi evoluindo desde 1985, até aos dias de hoje, continuando em exploração e em expansão para um melhor servir da população local. A existência de dois furos, e brevemente dum terceiro, servem para o abastecimento duma piscina, dum hotel, das termas, e da balneoterapia. Devido à riqueza a nível das temperaturas, dos caudais, e ao nível das necessidades energéticas existentes, este complexo apresenta um tempo de retorno de investimento de cerca de 7 anos, o que é geralmente considerado para investimentos para fins públicos, como é o caso. No âmbito das investigações agora realizadas, foi constatado que estes projetos suportam a cobertura de alguma incerteza hidrogeológica, dada a importante procura existente.

Redes de terras associadas a estações de telecomunicações

As redes de terras são algo que atormenta muitas operadoras de telecomunicações. Quando o comum civil pensa que apenas existem antenas e que é importante a transmissão de dados porque naturalmente não lhes faz falta saber mais nada, as pessoas envolventes na construção de uma estação de telecomunicações têm preocupações não só com a parte de rádio e antenas, mas fundamentalmente com a parte de energia e de infra-estruturas. No desenvolvimento deste projecto, torna-se então importante perceber como as redes de terras são implementadas, como atingir valores satisfatórios para as operadoras, como realizar melhoria da rede de terras já existentes, implementar furos artesianos/valas e sobretudo como melhorar no futuro. Os materiais utilizados e o estudo prévio das condições do terreno onde a estação rádio-base é implementada nem sempre coincidem com os resultados desejados e que pensamos que seriam facilmente atingidos após implementação da torre. Neste projecto, vamos então acompanhar todos os processos, desde a legalização, construção, testes e finalmente a medição da resistência de terras final. Se os valores foram aceitáveis, óptimo para a operadora de telecomunicações e para a empresa responsável pela implementação. Caso os resultados estejam aquém das expectativas, é altura de selecionar os melhores e recorrendo a um conjunto de técnicas que garantam resultados e o menos dispêndio de dinheiro possível, vamos avançar para as melhorias. Os regimes de protecção das pessoas e da estação são bastante importantes. É importante também perceber as condições dos solos e saber simular com o programa ERICO que será fundamental para se efetuar uma comparação entre o teórico e o implementado praticamente. No fim, parte fulcral serão as medições finais, a relação preço-qualidade das melhorias implementadas, a análise dos resultados com argumentos válidos e o bom senso da operadora para aceitar ou rejeitar os trabalhos. Contrato é contrato, fazer mais e melhor é uma exigência constante.

Reproduction of Leishmania (Leishmania ) infantum chagasi in conditioned cell culture growth medium

Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM). Leishmania (Leishmania) infantum chagasi (100 parasites) grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells) and its multiplication was observed for five days (120 hours). After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes). The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µ L and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes) used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times) as observed by growth kinetics, within a shorter time. These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias) or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits.

Investigações hidrogeológicas da sondagem CC3 do aquífero termomineral de Chaves: implicações para o modelo conceptual

O presente estudo centra‐se na conceptualização de pormenor do modelo de circulação do aquífero termomineral de Chaves na sua zona de descarga no campo hidromineral e geotérmico de Chaves (Norte de Portugal). Para tal, utilizam‐se as informações resultantes da bibliografia disponível, da construção das captações de água termomineral AC1 e AC2, já existentes, e, principalmente, as informações obtidas a partir dos trabalhos de pesquisa, prospecção e construção da captação CC3, obra desenvolvida no ano de 2014 para reforço de caudal das Caldas de Chaves. Incidindo fundamentalmente na análise de perfis geofísicos, de parâmetros de perfuração, “cuttings” e diagrafias diferidas inerentes à sondagem de pesquisa CC3, e da sua posterior transformação em captação, integrando‐se posteriormente toda a informação daí resultante, o estudo culmina na apresentação de um modelo conceptual de pormenor para a circulação de águas tipo “Chaves” na zona de descarga que permite, com considerável robustez, à luz dos dados conhecidos à presente data, o melhor conhecimento do aquífero para futuros projectos de desenvolvimento dos recursos hidrominerais e geotérmicos de Chaves.

Stratigraphic interpretation of Well-Log data of the Athabasca Oil Sands of Alberta Canada through Pattern recognition and Artificial Intelligence

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

QUANTITATIVE VS. CONVENTIONAL PCR FOR DETECTION OF HUMAN ADENOVIRUSES IN WATER AND SEDIMENT SAMPLES

SUMMARY Human Adenoviruses (HAdV) are notably resistant in the environment. These agents may serve as effective indicators of fecal contamination, and may act as causative agents of a number of different diseases in human beings. Conventional polymerase chain reaction (PCR) and, more recently, quantitative PCR (qPCR) are widely used for detection of viral agents in environmental matrices. In the present study PCR and SYBR(r)Green qPCR assays were compared for detection of HAdV in water (55) and sediments (20) samples of spring and artesian wells, ponds and streams, collected from dairy farms. By the quantitative methodology HAdV were detected in 87.3% of the water samples and 80% of the sediments, while by the conventional PCR 47.3% and 35% were detected in water samples and sediments, respectively.

WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL

SUMMARY The aim of this work was to compare, from a parasitological ( Cryptosporidiumspp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidiumspp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardiaspp. or Cryptosporidiumspp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidiumspp. and Giardiaspp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

Contribuição para a validação do código numérico OpenFOAM em escoamentos com superfície livre

Dissertação para obtenção do Grau de Mestre em Engenharia Mecânica

Two competitive enzyme immunoassays for the detection of IgG class antibodies to hepatitis a antigen

Two competitive enzyme immunoassays (EIA) techniques were developed: in the first (COMP-1), test sera were added together with HAV antigen on anti-HAV IgG-coated wells followed by an anti-HA VHRP conjugate; in the second (COMP-2), test sera and anti-HA VHRP conjugate competed for HAV epitopes previously adsorbed to anti-HA V IgG-coated wells. Both procedures used tetramethylbenzidine (TMB) as a substrate. Both competitive tests were shown to be reproducible and suitable for routine diagnosis and research purposes.

Caracterização da qualidade da energia entregue à rede por centrais de energia dos oceanos

Dissertação para obtenção do Grau de Mestre em Engenharia Electrotécnica e de Computadores

Correction of the anisotropy in resistivity: application to pore pressure prediction

Dissertação para obtenção do Grau de Mestre em Engenharia Geológica (Georrecursos)

Groundwater quality, vulnerability and potential assessment in Kobo Valley development project, Ethiopia

This study deals with investigating the groundwater quality for irrigation purpose, the vulnerability of the aquifer system to pollution and also the aquifer potential for sustainable water resources development in Kobo Valley development project. The groundwater quality is evaluated up on predicting the best possible distribution of hydrogeochemicals using geostatistical method and comparing them with the water quality guidelines given for the purpose of irrigation. The hydro geochemical parameters considered are SAR, EC, TDS, Cl-, Na+, Ca++, SO4 2- and HCO3 -. The spatial variability map reveals that these parameters falls under safe, moderate and severe or increasing problems. In order to present it clearly, the aggregated Water Quality Index (WQI) map is constructed using Weighted Arithmetic Mean method. It is found that Kobo-Gerbi sub basin is suffered from bad water quality for the irrigation purpose. Waja Golesha sub-basin has moderate and Hormat Golena is the better sub basin in terms of water quality. The groundwater vulnerability assessment of the study area is made using the GOD rating system. It is found that the whole area is experiencing moderate to high risk of vulnerability and it is a good warning for proper management of the resource. The high risks of vulnerability are noticed in Hormat Golena and Waja Golesha sub basins. The aquifer potential of the study area is obtained using weighted overlay analysis and 73.3% of the total area is a good site for future water well development. The rest 26.7% of the area is not considered as a good site for spotting groundwater wells. Most of this area fall under Kobo-Gerbi sub basin.

Localization in underground tunnels

Positioning technologies are becoming ubiquitous and are being used more and more frequently for supporting a large variety of applica- tions. For outdoor applications, global navigation satellite systems (GNSSs), such as the global positioning system (GPS), are the most common and popular choice because of their wide coverage. GPS is also augmented with network-based systems that exploit existing wireless and mobile networks for providing positioning functions where GPS is not available or to save energy in battery-powered devices. Indoors, GNSSs are not a viable solution, but many applications require very accurate, fast, and exible positioning, tracking, and navigation functions. These and other requirements have stim- ulated research activities, in both industry and academia, where a variety of fundamental principles, techniques, and sensors are being integrated to provide positioning functions to many applications. The large majority of positioning technologies is for indoor environments, and most of the existing commercial products have been developed for use in of ce buildings, airports, shopping malls, factory plants, and similar spaces. There are, however, other spaces where positioning, tracking, and navigation systems play a central role in safety and in rescue operations, as well as in supporting speci c activities or for scienti c research activities in other elds. Among those spaces are underground tunnels, mines, and even underwater wells and caves. This chapter describes the research efforts over the past few years that have been put into the development of positioning systems for underground tun- nels, with particular emphasis in the case of the Large Hadron Collider (LHC) at CERN (the European Organization for Nuclear Research), where localiza- tion aims at enabling more automatic and unmanned radiation surveys. Examples of positioning and localization systems that have been devel- oped in the past few years for underground facilities are presented in the fol- lowing section, together with a brief characterization of those spaces’ special conditions and the requirements of some of the most common applications. Section 5.2 provides a short overview of some of the most representative research efforts that are currently being carried out by many research teams around the world. In addition, some of the fundamental principles and tech- niques are identi ed, such as the use of leaky coaxial cables, as used at the LHC. In Section 5.3, we introduce the speci c environment of the LHC and de ne the positioning requirements for the envisaged application. This is followed by a detailed description of our approach and the results that have been achieved so far. Some last comments and remarks are presented in a nal section.

Metodologias para a qualificação de águas minerais naturais e águas de nascente

Dissertação de mestrado em Ordenamento e Valorização de Recursos Geológicos