Late Quaternary warm marine mollusks from Santa Maria (Azores); paleoecologic and paleobiogeographic considerations

The sub-fossil fauna from the Late Quaternary marine deposits of Santa Maria is made of more than 50 species of gastropods and bivalves, 19 of them collected recently and for the first time in the northern coast of the island (Lagoinhas Bay). The sub-fossil shells are found in deposits of beach sands, situated 2-3 meters above the present low tide. The carbonated sands from the basal part of the succession yield an autochthonous association of borers dominated by the bivalve Myoforceps aristata (Dillwin, 1817). Upwards, the marine sands contain concentrations of beach drift shells, including well-preserved supratidal and intertidal gastropods, among them a large number of Rissoidae. The bivalve fauna is dominated by disarticulated valves of Ervilia castanea (Montagu, 1803), a small infaunal coloniser of mobile sandy substrates. The composition of the fauna is made essentially of West European species, many of them common to the West Coast of Portugal. However, a few "warm guests" with West African or Caribbean affinities were also found, suggesting a close relation with some of the "Tyrrhenian" warm associations found in the Western Mediterranean.

Simple laccase-based biosensor for formetanate hydrochloride quantification in fruits

This work describes the development of an electrochemical enzymatic biosensor for quantification of the pesticide formetanate hydrochloride (FMT). It is based on a gold electrode modified with electrodeposited gold nanoparticles and laccase. The principle behind its development relies on FMT's capacity to inhibit the laccase catalytic reaction that occurs in the presence of phenolic substrates. The optimum values for the relevant experimental variables such as gold nanoparticles electrochemical deposition (at − 0.2 V for 100 s), laccase immobilization (via glutaraldehyde cross-linking), laccase concentration (12.4 mg/mL), substrate selection and concentration (5.83×10−5 M of aminophenol), pH (5.0), buffer (Britton–Robinson), and square-wave voltammetric parameters were determined. The developed biosensor was successfully applied to FMT determination in mango and grapes. The attained limit of detection was 9.5×10−8 ± 9.5×10−10 M (0.02 ± 2.6×10−4 mg/kg on a fresh fruit weight basis). Recoveries for the five tested spiking levels ranged from 95.5 ± 2.9 (grapes) to 108.6 ± 2.5% (mango). The results indicated that the proposed device presents suitable characteristics in terms of sensitivity (20.58 ± 0.49 A/μM), linearity (9.43×10−7 to 1.13×10−5 M), accuracy, repeatability (RSD of 1.4%), reproducibility (RSD of 1.8%) and stability (19 days) for testing of compliance with established maximum residue limits of FMT in fruits and vegetables.

Solution-Processable Donor-Acceptor-Donor Oligomers with Cross-Linkable Functionality

Electron-acceptor units, combined with bithiophene substituted with flexible chains end-functionalized with cross-linkable moieties, provide soluble donor-acceptor-donor (DAD) it-conjugated oligomer-type molecules with cross-linking ability and broad absorption in the visible spectrum. A study on the cross-linking conditions of the new oligomers to yield insoluble polymer networks is presented, including conditions for obtaining polymer films over poly(3,4-ethylenedioxythiophene):polystyrene sulfonate-covered substrates. The combination of the DAD molecular design and cross-linking functionality opens prospects for applications in solution-processed small-molecule solar cells with morphologically-stable organic layers.

Influence of the substrate on structure and magnetic properties of Co-N thin films

Cubic cobalt nitride films were grown onto different single crystalline substrates Al2O3 (0 0 0 1) and (1 1 View the MathML source 0), MgO (1 0 0) and (1 1 0) and TiO2 (1 0 0) and (1 1 0). The films display low atomic densities compared with the bulk material, are ferromagnetic and have metallic electrical conductivity. X-ray diffraction and X-ray absorption fine structure confirm the cubic structure of the films and with RBS results indicate that samples are not homogeneous at the microscopic scale, coexisting Co4+xN nitride with nitrogen rich regions. The magnetization of the films decreases with increase of the nitrogen content, variation that is shown to be due to the decrease of the cobalt density, and not to a decrease of the magnetic moment per cobalt ion. The films are crystalline with a nitrogen deficient stoichiometry and epitaxial with orientation determined by the substrate.

The role of the efflux mechanisms in multidrug resistance in Mycobacterium tuberculosis

Abstract The emergence of multi and extensively drug resistant tuberculosis (MDRTB and XDRTB) has increased the concern of public health authorities around the world. The World Health Organization has defined MDRTB as tuberculosis (TB) caused by organisms resistant to at least isoniazid and rifampicin, the main first-line drugs used in TB therapy, whereas XDRTB refers to TB resistant not only to isoniazid and rifampicin, but also to a fluoroquinolone and to at least one of the three injectable second-line drugs, kanamycin, amikacin and capreomycin. Resistance in Mycobacterium tuberculosis is mainly due to the occurrence of spontaneous mutations and followed by selection of mutants by subsequent treatment. However, some resistant clinical isolates do not present mutations in any genes associated with resistance to a given antibiotic, which suggests that other mechanism(s) are involved in the development of drug resistance, namely the presence of efflux pump systems that extrude the drug to the exterior of the cell, preventing access to its target. Increased efflux activity can occur in response to prolonged exposure to subinhibitory concentrations of anti-TB drugs, a situation that may result from inadequate TB therapy. The inhibition of efflux activity with a non-antibiotic inhibitor may restore activity of an antibiotic subject to efflux and thus provide a way to enhance the activity of current anti-TB drugs. The work described in this thesis foccus on the study of efflux mechanisms in the development of multidrug resistance in M. tuberculosis and how phenotypic resistance, mediated by efflux pumps, correlates with genetic resistance. In order to accomplish this goal, several experimental protocols were developed using biological models such as Escherichia coli, the fast growing mycobacteria Mycobacterium smegmatis, and Mycobacterium avium, before their application to M. tuberculosis. This approach allowed the study of the mechanisms that result in the physiological adaptation of E. coli to subinhibitory concentrations of tetracycline (Chapter II), the development of a fluorometric method that allows the detection and quantification of efflux of ethidium bromide (Chapter III), the characterization of the ethidium bromide transport in M. smegmatis (Chapter IV) and the contribution of efflux activity to macrolide resistance in Mycobacterium avium complex (Chapter V). Finally, the methods developed allowed the study of the role of efflux pumps in M. tuberculosis strains induced to isoniazid resistance (Chapter VI). By this manner, in Chapter II it was possible to observe that the physiological adaptation of E. coli to tetracycline results from an interplay between events at the genetic level and protein folding that decrease permeability of the cell envelope and increase efflux pump activity. Furthermore, Chapter III describes the development of a semi-automated fluorometric method that allowed the correlation of this efflux activity with the transport kinetics of ethidium bromide (a known efflux pump substrate) in E. coli and the identification of efflux inhibitors. Concerning M. smegmatis, we have compared the wild-type M. smegmatis mc2155 with knockout mutants for LfrA and MspA for their ability to transport ethidium bromide. The results presented in Chapter IV showed that MspA, the major porin in M. smegmatis, plays an important role in the entrance of ethidium bromide and antibiotics into the cell and that efflux via the LfrA pump is involved in low-level resistance to these compounds in M. smegmatis. Chapter V describes the study of the contribution of efflux pumps to macrolide resistance in clinical M. avium complex isolates. It was demonstrated that resistance to clarithromycin was significantly reduced in the presence of efflux inhibitors such as thioridazine, chlorpromazine and verapamil. These same inhibitors decreased efflux of ethidium bromide and increased the retention of [14C]-erythromycin in these isolates. Finaly, the methods developed with the experimental models mentioned above allowed the study of the role of efflux pumps on M. tuberculosis strains induced to isoniazid resistance. This is described in Chapter VI of this Thesis, where it is demonstrated that induced resistance to isoniazid does not involve mutations in any of the genes known to be associated with isoniazid resistance, but an efflux system that is sensitive to efflux inhibitors. These inhibitors decreased the efflux of ethidium bromide and also reduced the minimum inhibitory concentration of isoniazid in these strains. Moreover, expression analysis showed overexpression of genes that code for efflux pumps in the induced strains relatively to the non-induced parental strains. In conclusion, the work described in this thesis demonstrates that efflux pumps play an important role in the development of drug resistance, namely in mycobacteria. A strategy to overcome efflux-mediated resistance may consist on the use of compounds that inhibit efflux activity, restoring the activity of antimicrobials that are efflux pump substrates, a useful approach particularly in TB where the most effective treatment regimens are becoming uneffective due to the increase of MDRTB/XDRTB.

Energy evaporation: the new concept of indoor systems for WPT and EH embedded into the floor

This work introduces a novel idea for wireless energy transfer, proposing for the first time the unit-cell of an indoor localization and RF harvesting system embedded into the floor. The unit-cell is composed by a 5.8 GHz patch antenna surrounded by a 13.56 MHz coil. The coil locates a device and activate the patch which, connected to a power grid, radiates to wirelessly charge the localized device. The HF and RF circuits co-existence and functionality are demonstrated in this paper, the novelty of which is also in the adoption of low cost and most of all ecofriendly materials, such as wood and cork, as substrates for electronics.

Hydrogen evolution on nanostructured Ni-Cu foams

Three-dimensional (3D) nickel-copper (Ni-Cu) nanostructured foams were prepared by galvanostatic electrodeposition, on stainless steel substrates, using the dynamic hydrogen bubble template. These foams were tested as electrodes for the hydrogen evolution reaction (HER) in 8 M KOH solutions. Polarisation curves were obtained for the Ni-Cu foams and for a solid Ni electrode, in the 25-85 degrees C temperature range, and the main kinetic parameters were determined. It was observed that the 3D foams have higher catalytic activity than pure Ni. HER activation energies for the Ni-Cu foams were lower (34-36 kJ mol(-1)) than those calculated for the Ni electrode (62 kJ mol(-1)). The foams also presented high stability for HER, which makes them potentially attractive cathode materials for application in industrial alkaline electrolysers.

Dual-phase inkjet printed electrochromic layers based on PTA and WOX/TiO2 nanoparticles for electrochromic applications

Thesis submitted in Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa for the degree of Master in Materials Engineering

Molecular details of INH-C-10 binding to wt KatG and Its S315T mutant

Isoniazid (INH) is still one of the two most effective antitubercular drugs and is included in all recommended multitherapeutic regimens. Because of the increasing resistance of Mycobacterium tuberculosis to INH, mainly associated with mutations in the katG gene, new INH-based compounds have been proposed to circumvent this problem. In this work, we present a detailed comparative study of the molecular determinants of the interactions between wt KatG or its S315T mutant form and either INH or INH-C10, a new acylated INH derivative. MD simulations were used to explore the conformational space of both proteins, and results indicate that the S315T mutation did not have a significant impact on the average size of the access tunnel in the vicinity of these residues. Our simulations also indicate that the steric hindrance role assigned to Asp137 is transient and that electrostatic changes can be important in understanding the enzyme activity data of mutations in KatG. Additionally, molecular docking studies were used to determine the preferred modes of binding of the two substrates. Upon mutation, the apparently less favored docking solution for reaction became the most abundant, suggesting that S315T mutation favors less optimal binding modes. Moreover, the aliphatic tail in INH-C10 seems to bring the hydrazine group closer to the heme, thus favoring the apparent most reactive binding mode, regardless of the enzyme form. The ITC data is in agreement with our interpretation of the C10 alkyl chain role and helped to rationalize the significantly lower experimental MIC value observed for INH-C10. This compound seems to be able to counterbalance most of the conformational restrictions introduced by the mutation, which are thought to be responsible for the decrease in INH activity in the mutated strain. Therefore, INH-C10 appears to be a very promising lead compound for drug development.

Influência do tipo de adesivo em juntas híbridas soldadas-adesivas

A necessidade de utilizar métodos de ligação entre componentes de forma mais rápida, eficaz e com melhores resultados, tem causado a crescente utilização das juntas adesivas, em detrimento dos métodos tradicionais de ligação tais como a soldadura, brasagem, ligações aparafusadas e rebitadas. A utilização das juntas adesivas tem vindo a aumentar em diversas aplicações industriais por estas apresentarem vantagens das quais se destacam a redução de peso, redução de concentrações de tensões e facilidade de fabrico. No entanto, também apresentam desvantagens, como a necessidade de preparação das juntas e o descentramento da carga aplicada que provoca efeitos de flexão, os quais dão origem a tensões normais na direcção da espessura do adesivo (tensões de arrancamento), afectando assim a resistência da junta. A combinação da ligação adesiva com a soldadura por pontos permite algumas vantagens em comparação com as juntas adesivas tradicionais como a maior resistência, aumento da rigidez, melhor resistência ao corte e arrancamento e também à fadiga. Neste trabalho é apresentado um estudo experimental e numérico de juntas de sobreposição simples adesivas e híbridas (adesivas-soldadas). Os adesivos utilizados são o Araldite AV138®, apresentado como sendo frágil, e os adesivos Araldite 2015® e Sikaforce® 7752, intitulados como adesivos dúcteis. Foram considerados substratos de aço (C45E) em juntas com diferentes comprimentos de sobreposição ( ), que foram sujeitas a esforços de tracção. Foi realizada uma análise dos valores experimentais e efectuada uma comparação destes valores com os resultados obtidos por Elementos Finitos (EF) no software ABAQUS®, que incluiu uma análise de tensões na camada de adesivo e previsão do comportamento das juntas por Modelos de Dano Coesivo (MDC). A análise por MDC permitiu obter os modos de rotura, as curvas força-deslocamento e a resistência das juntas com bastante precisão, com excepção das juntas coladas com o adesivo Sikaforce® 7752. Estes resultados permitiram validar a técnica de modelação proposta para as juntas coladas e híbridas, o que representa uma base para posterior aplicação desta técnica em projecto, com as vantagens decorrentes da redução do tempo de projecto e maior facilidade de optimização.

Ligação Adesiva de Polímeros Particularmente Difíceis de Colar

As ligações adesivas têm sido cada vez mais utilizadas nos últimos anos em detrimento de outros métodos tais como a soldadura, ligações aparafusadas e ligações rebitadas. Os plásticos de Engenharia têm um papel cada vez mais preponderante na indústria, devido às suas excelentes propriedades. Neste trabalho foram considerados três polímeros diferentes, o Policloreto de Vinilo (PVC) e o Polipropileno (PP) dado o seu baixo custo e peso e a superfície quimicamente inerte e o Politetrafluoretileno (PTFE) devido às suas boas propriedades químicas e excelentes propriedades de deslizamento. No entanto, estes materiais possuem uma baixa energia de superfície e, por isso, são muito difíceis de colar com mais relevância para o PTFE. Assim, após um estudo preliminar foi escolhido, para realizar as colagens necessárias, um adesivo da Tamarron Technology “Tam Tech Adhesive”, próprio para este tipo de substratos difíceis de colar. Posteriormente foi efetuada a sua caraterização através de ensaios de provetes maciços à tração. O principal objetivo deste trabalho foi estudar juntas de sobreposição simples de materiais poliméricos difíceis de colar tais como o PTFE, PP e PVC com recurso a um adesivo que não necessitasse de preparação de superfície. Foram fabricadas juntas de sobreposição simples (JSS) segundo os métodos Lap Shear (LS) e Block Shear (BS) dos três materiais referidos anteriormente e realizados os respetivos ensaios para avaliar o comportamento mecânico das ligações adesivas. Os materiais utilizados como substratos foram também submetidos a ensaios de tração com a finalidade de obter o módulo de elasticidade e as suas propriedades de resistência. Os substratos envolvidos nas juntas adesivas não sofreram qualquer preparação especial das superfícies. Na maioria dos casos consistiu apenas numa limpeza das superfícies com álcool etílico. Contudo, para o PTFE também se experimentou a preparação por abrasão com lixa e por chama. Foi também efetuado um trabalho de simulação numérica por elementos finitos utilizando um modelo de dano coesivo triangular. As resistências ao corte obtidas são superiores em BS comparativamente a LS, exceção feita aos substratos de PTFE aonde os resultados são similares. O tratamento por chama melhorou a resistência mecânica das juntas. Verificou-se também que o modelo numérico simulou adequadamente o comportamento das juntas principalmente das LS.

Light controlled synthesis of nucleic acids

Dissertação apresentada para obtenção do grau de Doutor em Bioquímica - especialidade Biotecnologia, pela Universidade Nova de Lisboa,Faculdade de Ciências e Tecnologia

Development of new biomaterials based on liquid crystalline phases of cellulose derivatives

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do Grau de Mestre em Engenharia Biomédica

The transport of carboxylic acids and important role of the Jen1p transporter during the development of yeast colonies

On solid substrates, yeast colonies pass through distinct developmental phases characterized by the changes in pH of their surroundings from acidic to nearly alkaline and vice versa. At the beginning of the alkali phase colonies start to produce ammonia, which functions as a quorum-sensing molecule inducing the reprogramming of cell metabolism. Such reprogramming includes, among others, the activation of several plasma membrane transporters and is connected with colony differentiation. In the present study, we show that colony cells can use two transport mechanisms to import lactic acid: a ‘saturable’ component of the transport, which requires the presence of a functional Jen1p transporter, and a ‘non-saturable’ component (diffusion) that is independent of Jen1p. During colony development, the efficiency of both transport components changes similarly in central and outer colonial cells. Although the lactate uptake capacity of central cells gradually decreases during colony development, the lactate uptake capacity of outer cells peaks during the alkali phase and is also kept relatively high in the second acidic phase. This lactate uptake profile correlates with the localization of the Jen1p transporter to the plasma membrane of colony cells. Both lactic acid uptake mechanisms are diminished in sok2 colonies where JEN1 expression is decreased. The Sok2p transcription factor may therefore be involved in the regulation of non-saturable lactic acid uptake in yeast colonies.

Neural mechanisms of stimulus generalization in auditory fear conditioning

Dissertation presented to obtain the Ph.D degree in Neuroscience Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa