Thermodynamic and kinetic modelling of the redox properties of tetrahaem cytochromes C3

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

Tailoring of structures and permeation properties of asymmetric nanocomposite cellulose acetatesilver membranes

Cellulose acetate (CA)-silver (Ag) nanocomposite asymmetric membranes were prepared via the wet-phase inversion method by dispersing polyvinylpirrolydone-protected Ag nanoparticles in the membrane casting solutions of different compositions. Silver nanoparticles were synthesized ex situ and added to the casting solution as a concentrated aqueous colloidal dispersion. The effects of the dispersion addition on the structure and on the selective permeation properties of the membranes were studied by comparing the nanocomposites with the silver-free materials. The casting solution composition played an important role in the adequate dispersion of the silver nanoparticles in the membrane. Incorporation of nanoscale silver and the final silver content resulted in structural changes leading to an increase in the hydraulic permeability and molecular weight cut-off of the nanocomposite membranes. (c) 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015, 132, 41796.

Alternative chelating agents: Evaluation of the ready biodegradability and complexation properties

The ready biodegradability of four chelating agents, N,N -(S,S)-bis[1-carboxy-2-(imidazol-4-yl)ethyl]ethylenediamine (BCIEE), N - ethylenedi-L-cysteine (EC), N,N -bis (4-imidazolymethyl)ethylenediamine (EMI) and 2,6-pyridine dicarboxylic acid (PDA), was tested according to the OECD guideline for testing of chemicals. PDA proved to be a readily biodegradable substance. However, none of the other three compounds were degraded during the 28 days of the test. Chemical simulations were performed for the four compounds in order to understand their ability to complex with some metal ions (Ca, Cd, Co, Cu, Fe, Mg, Mn, Ni, Pb, Zn) and discuss possible applications of these chelating agents. Two different conditions were simulated: (i) in the presence of the chelating agent and one metal ion, and (ii) in the simultaneous presence of the chelating agent and all metal ions with an excess of Ca. For those compounds that were revealed not to be readily biodegradable (BCIEE, EC and EMI), applications were evaluated where this property was not fundamental or even not required. Chemical simulations pointed out that possible applications for these chelating agents are: food fortification, food process, fertilizers, biocides, soil remediation and treatment of metal poisoning. Additionally, chemical simulations also predicted that PDA is an efficient chelating agent for Ca incrustations removal, detergents and for pulp metal ions removal process.

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.

Antibacterial activity of Ionic Liquids based on ampicillin against resistant bacteria

Antibacterial activity of novel Active Pharmaceutical Ingredient Ionic Liquids (API-ILs) based on ampicillin anion [Amp] have been evaluated. They showed growth inhibition and bactericidal properties on some sensitive bacteria and especially some Gram-negative resistant bacteria when compared to the [Na][Amp] and the initial bromide and chloride salts. For these studies were analysed the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBIC) against sensitive Gram-negative bacteria Escherichia coli ATCC 25922 and Klebsiella pneumonia (clinically isolated), as well as sensitive Gram positive S. Aureus ATCC 25923, Staphylococcus epidermidis and Enterococcus faecalis and completed using clinically isolated resistent strains: E. coli TEM CTX M9, E. coli CTX M2 and E. coli AmpC Mox. From the obtained MIC values of studied APIs-ILs and standard [Na][Amp] were derived RDIC values (relative decrease of inhibitory concentration). High RDIC values of [C16Pyr][Amp] especially against two resistant Gram-negative strains E. coli TEM CTX M9 (RDIC>1000) and E. coli CTX M2 (RDIC>100) point clearly to a potential promising role of APIs-ILs as antimicrobial drugs especially against resistant bacterial strains.

Serological, electrophoretic and biological properties of Fasciola hepatica antigens

Fasciola hepatica somatic antigen, its partially purified fractions and excretion-secretion products were investigated as to serological, electrophoretic and biological properties. In a Sephadex G-100 column (SG-100), Fasciola hepatica total antigen (FhTA) gave 5 fractions, and SDS-PAGE analysis showed they were glycoproteins ranging from 14 to 94 kDa molecular weight (MW). When these fractions were analyzed by enzyme linked immunotransfer blot (EITB) and immunodiffusion in gel (ID) with serum from immunized rats with FhTA, the presence of different antigenic components was revealed. In the SDS-PAGE of excretor-secretor antigen (ESA), it was possible to observe peptides from 12 to 22 kDa, which were also present in FhTA. When the FhTA, its fractions and the ESA were analyzed by EITB with the immune rat serum (IRS), it was observed that only some fractions of the SG-100 shared antigens with the FhTA and ESA. Moreover, DTH and ITH responses were studied in FhTA immunized rats challenged with these different antigen components, revealing that the protein/carbohydrate ratio is important for inducing DTH response. The ESA was the most active component in the DTH and ITH response.

Low temperature structural transitions in dipolar hard spheres: the influence on magnetic properties

We investigate the structural chain-to-ring transition at low temperature in a gas of dipolar hard spheres (DRS). Due to the weakening of entropic contribution, ring formation becomes noticeable when the effective dipole-dipole magnetic interaction increases, It results in the redistribution of particles from usually observed flexible chains into flexible rings. The concentration (rho) of DI-IS plays a crucial part in this transition: at a very low rho only chains and rings are observed, whereas even a slight increase of the volume fraction leads to the formation of branched or defect structures. As a result, the fraction of DHS aggregated in defect-free rings turns out to be a non-monotonic function of rho. The average ring size is found to be a slower increasing function of rho when compared Lo that of chains. Both theory and computer simulations confirm the dramatic influence of the ring formation on the rho-dependence of the initial magnetic susceptibility (chi) when the temperature decreases. The rings clue to their zero total dipole moment are irresponsive to a weak magnetic field and drive to the strong decrease of the initial magnetic susceptibility. (C) 2014 Elsevier B.V. All rights reserved.

Antimicrobial resistance and plasmid detection in strains of the Bacteroides fragilis group

Resistant populations of the Bacteroides fragilis group bacteria (two reference ones and two isolated from human and Callithrix penicillata marmoset) were obtained by the gradient plate technique, to clindamycin, penicillin G, metronidazole and mercuric chloride. All the four tested strains were originaly susceptible to the four antimicrobial drugs at the breakpoint used in this study. MICs determination for the four cultures gave constant values for each antimicrobial, on the several steps by the gradient plate technique. The intestinal human B. fragilis strains showed three DNA bands, that could be representative of only two plasmids in the closed covalently circular (CCC) form with molecular weights of approximately 25 and 2.5 Md. The results do not permit an association between the presence of plasmid in the human strain with the susceptibility to the studied drugs. The four strains were ß-lactamase negative in the two methods used, and no particular chromosomal genetic resistance marker was demonstred. The resistance (MIC) observed, after contact with penicillin G and mercuric chloride, were two-fold in the four tested strains

Estimation of the Crustal Bulk Properties Beneath Mainland Portugal from P-Wave Teleseismic Receiver Functions

In this work, we present results from teleseismic P-wave receiver functions (PRFs) obtained in Portugal, Western Iberia. A dense seismic station deployment conducted between 2010 and 2012, in the scope of the WILAS project and covering the entire country, allowed the most spatially extensive probing on the bulk crustal seismic properties of Portugal up to date. The application of the H-κ stacking algorithm to the PRFs enabled us to estimate the crustal thickness (H) and the average crustal ratio of the P- and S-waves velocities V p/V s (κ) for the region. Observations of Moho conversions indicate that this interface is relatively smooth with the crustal thickness ranging between 24 and 34 km, with an average of 30 km. The highest V p/V s values are found on the Mesozoic-Cenozoic crust beneath the western and southern coastal domain of Portugal, whereas the lowest values correspond to Palaeozoic crust underlying the remaining part of the subject area. An average V p/V s is found to be 1.72, ranging 1.63-1.86 across the study area, indicating a predominantly felsic composition. Overall, we systematically observe a decrease of V p/V s with increasing crustal thickness. Taken as a whole, our results indicate a clear distinction between the geological zones of the Variscan Iberian Massif in Portugal, the overall shape of the anomalies conditioned by the shape of the Ibero-Armorican Arc, and associated Late Paleozoic suture zones, and the Meso-Cenozoic basin associated with Atlantic rifting stages. Thickened crust (30-34 km) across the studied region may be inherited from continental collision during the Paleozoic Variscan orogeny. An anomalous crustal thinning to around 28 km is observed beneath the central part of the Central Iberian Zone and the eastern part of South Portuguese Zone.

Properties and Sustainability of Biodiesel from Animal Fats and Fish Oil

This work presents and analyses the fat and fuel properties and the methyl ester profile of biodiesel from animal fats and fish oil (beef tallow, pork lard, chicken fat and sardine oil). Also, their sustainability is evaluated in comparison with rapeseed biodiesel and fossil diesel, currently the dominant liquid fuels for transportation in Europe. Results show that from a technological point of view it is possible to use animal fats and fish oil as feedstock for biodiesel production. From the sustainability perspective, beef tallow biodiesel seems to be the most sustainable one, as its contribution to global warming has the same value of fossil diesel and in terms of energy efficiency it has the best value of the biodiesels under consideration. Although biodiesel is not so energy efficient as fossil diesel there is room to improve it, for example, by replacing the fossil energy used in the process with renewable energy generated using co-products (e.g. straw, biomass cake, glycerine).

Influence of the crushing process of recycled aggregates on concrete properties

This work intends to evaluate the (mechanical and durability) performance of concrete made with coarse recycled concrete aggregates (CRCA) obtained using two crushing processes: primary crushing (PC) and primary plus secondary crushing (PSC). This analysis intends to select the most efficient production process of recycled aggregates (RA). The RA used here resulted from precast products (P), with strength classes of 20 MPa, 45 MPa and 65 MPa, and from laboratory-made concrete (L) with the same compressive strengths. The evaluation of concrete was made with the following tests: compressive strength; splitting tensile strength; modulus of elasticity; carbona-tion resistance; chloride penetration resistance; capillary water absorption; and water absorption by immersion. These findings contribute to a solid and innovative basis that allows the precasting industry to use without restrictions the waste it generates. © (2015) Trans Tech Publications, Switzerland.

Unravelling novel modes of antimicrobial action

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

Bioatividade de Líquidos Iónicos derivados do Ácido Valpróico em bactérias Gram-negativa

Os Líquidos Iónicos (LIs) são sais orgânicos constituídos exclusivamente por iões e possuem pontos de fusão inferiores a 100ºC. As suas propriedades únicas e o facto de ser possível ajustar as suas propriedades físicas, químicas e biológicas, de acordo com o objetivo pretendido, tornam esta classe de compostos, um grande objeto de estudo de inúmeros investigadores. Desde os inícios da sua aplicação até à atualidade, a investigação nesta área expandiu o seu raio de ação, estando já descrito o seu potencial como agentes antimicrobianos e, mais recentemente, como compostos farmacêuticos ativos. Atualmente muitas das suas aplicações são baseadas nas suas propriedades biológicas. Esta Tese teve como objetivo avaliar a influência que os LIs podem exercer a nível do crescimento bacteriano e estudar alternativas de combater a resistência bacteriana. Todos os LIs utilizados neste trabalho tinham como anião o ácido valpróico, sendo utilizados catiões orgânicos de amónio e de imidazólio. Foram utilizadas 4 bactérias e avaliou-se a atividade biológica e a respetiva taxa de crescimento. O estudo da sua atividade biológica foi feito através da determinação da Concentração Mínima Inibitória (CMI) e a análise das suas curvas de crescimentos na presença e ausência de composto. Com este trabalho foi possível verificar que dentro dos compostos em estudo, LIs derivados do valproato, o Valproato com o cetilperidínio [valp] [cetylpir] foi o que influenciou o crescimento de todas as bactérias estudadas. Este estudo demonstrou o potencial antibacteriano de alguns compostos, podendo desta forma vir a ser utilizados para fins farmacêuticos

Doseamento da Azadiractina e avaliação da atividade antimicrobiana em produtos contendo óleo de Neem

O Neem (Azadirachta indica) é uma árvore indiana conhecida pela atividade pesticida e por várias atividades farmacológicas. De entre os vários compostos já isolados e estudados, a Azadiractina (AZA) foi identificada como o principal composto bioativo desta planta. Este composto apresenta uma grande diversidade de localizações nesta planta, porém assume a sua máxima concentração ao nível das sementes, porção que se apresenta também como a principal fonte de obtenção do óleo de Neem. O óleo apresenta-se como a porção menos estudada do Neem, quer ao nível do seu teor em AZA, quer ao nível das suas propriedades, nomeadamente antimicrobianas. Neste sentido, os objetivos primordiais deste estudo foram o doseamento da Azadiractina e a avaliação da atividade antimicrobiana em produtos contendo óleo de Neem. Um método analítico rápido, sensível e seletivo utilizando HPLC-UV foi desenvolvido para a identificação e quantificação da Azadiractina-A (AZA-A) e 3-tigloylazadirachtol (AZA-B) em diferentes amostras de óleo de Neem. O teor de AZA-A, B e A+B determinado nas amostras de óleo de Neem apresentou valores entre 58,53-843,42 mg/kg, 12,52-800,223 mg/kg e 104,20-1642,17 mg/kg, respetivamente. Na generalidade, os valores obtidos foram inferiores aos descritos na literatura. A partir dos resultados obtidos, verificou-se ainda que o teor destes compostos não é similar em todas as amostras, sendo este condicionado pela qualidade das sementes que deram origem ao óleo e pelo processo extrativo utilizado. Para além disso, foi possível inferir que duas das amostras testadas teriam qualidade inferior, dados os teores reduzidos de AZA que apresentavam. As diferentes amostras de óleo de Neem, bem como formulações comerciais contendo óleo de Neem, foram testadas em 14 microrganismos de forma a avaliar o seu potencial antimicrobiano. Após a análise, verificou-se atividade antimicrobiana de todas as amostras sobre todos os microrganismos testados, observando-se atividade tanto em bactérias Gram+ como Gram-. Os resultados alcançados mostraram que o óleo de Neem e as formulações comerciais contendo óleo de Neem têm um potencial antimicrobiano interessante, principalmente sobre bactérias comuns em patologias da pele. Para além disso, foi possível comprovar que, no caso do óleo de Neem, a AZA não será a principal responsável por esta atividade. Por outro lado, verificou-se que a atividade antimicrobiana das formulações comerciais não se deverá exclusivamente à presença do óleo de Neem, Doseamento da Azadiractina e avaliação da atividade antimicrobiana em produtos contendo óleo de Neem X uma vez que os valores dos halos de inibição obtidos com as formulações tenderam a ser superiores aos verificados apenas com o óleo, além de que os valores de inibição mais elevados foram observados para as formulações contendo menor percentagem de óleo de Neem incorporado. Em suma, os resultados alcançados para os diferentes produtos analisados são promissores e, na sua maioria, convergem com o que está descrito na literatura. No entanto, apesar destes resultados serem um grande contributo, mais estudos são necessários e importantes para conhecer melhor os produtos analisados e assim poder tirar o maior proveito deles.

Stress Ratio Effects on the fatigue Properties of Biaxially Loaded Tubular FRP-Specimens – Experimental Study

In this work tubular fiber reinforced specimens are tested for fatigue life. The specimens are biaxially loaded with tension and shear stresses, with a load angle β of 30° and 60° and a load ratio of R=0,1. There are many factors that affect fatigue life of a fiber reinforced material and the main goal of this work is to study the effects of load ratio R by obtaining S-N curves and compare them to the previous works (1). All the other parameters, such as specimen production, fatigue loading frequency and temperature, will be the same as for the previous tests. For every specimen, stiffness, temperature of the specimen during testing, crack counting and final fracture mode are obtained. Prior to testing, a study if the literature regarding the load ratio effects on composites fatigue life and with that review estimate the initial stresses to be applied in testing. In previous works (1) similar specimens have only been tested for a load ratio of R=-1 and therefore the behaviour of this tubular specimens for a different load ratio is unknown. All the data acquired will be analysed and compared to the previous works, emphasizing the differences found and discussing the possible explanations for those differences. The crack counting software, developed at the institute, has shown useful before, however different adjustments to the software parameters lead to different cracks numbers for the same picture, and therefore a better methodology will be discussed to improve the crack counting results. After the specimen’s failure, all the data will be collected and stored and fibre volume content for every specimen is also determinate. The number of tests required to make the S-N curves are obtained according to the existent standards. Additionally are also identified some improvements to the testing machine setup and to the procedures for future testing.