RF-PECVD was used to prepare amorphous of carbon (DLC) onto stainless steel 316 and glass substrates. The substrates were negatively biased at between 100 V to 400 V. Thin films of DLC have been deposited using C2H2 and titanium isopropoxide (TIPOT). Argon was used to generate the plasma in the PECVD system chamber. DEKTAK 8 surface stylus profilometer was used to measure the film thickness and the deposition rate was calculated. Micro Raman spectroscopy was employed to determine the chemical structure and bonding present in the films. Composition analysis of the samples was carried out using VGTOF SIMS (IX23LS) instrument. In addition, X-ray photoelectron spectroscopy (XPS) was used to analyze the composition and chemical state of the films. The wettability of the films was examined using the optical contact angle meter (CAM200) system. Two types of liquids with different polarities were used to study changes in the surface energy. The as-grown films were in the thickness range of 200-400 nm. Raman spectroscopy results showed that the I(D)/I(G) ratio decreased when the bias voltage on the stainless steel substrates was increased. This indicates an increase in the graphitic nature of the film deposited. In contrast, on the glass substrates the I(D)/I(G) ratio increased when the bias voltage was increased indicates a greater degree of diamond like character. Chemical composition determined using XPS showed the presence of carbon and oxygen in both samples on glass and stainless steel substrates. Both coatings the contact angle of the films decreased except for 400 V which showed a slight increase. The oxygen is thought to play an important role on the polar component of a-C.

Time-of-flight secondary ion mass spectrometry was used to study fourhumancalculi and to compare the results with those from twelve commercially available urinary calculi minerals including three organic compounds (L-cystine, uric acid and sodium urate). Phase identification of calcium phos- phate compounds was carried out by considering the relative ion abundances of [Ca2O]R and [CaPO2]R. Deprotonated [M–H] and protonated [MRH]R uric acid were detected and used for component recognition in pure uric acid and in the mixed samples of struvite, calcium oxalate and uric acid. Iodine related to the medical history of a patient was also detected.

This work demonstrates experimentally the use of mass spectrometry (MS) for on-line, quantitative monitoring of organophilic pervaporation processes operated not only under variable conditions of tem- perature of condensation/downstream pressure but also when using complex multi-component feed streams. Due to its high sensitivity, mass spectrometry is particularly suitable for on-line monitoring of aromas in dilute streams under reduced pressure, as happens with natural aromas in permeate streams. Mass spectrometry is also suitable for on-line monitoring of solvent and co-solvent permeants (water and ethanol in the selected case study), enabling for on-line determination of selectivity of the solutes of interest. A new method of calibration is proposed and validated, which correlates the intensity of the characteristic mass peak mi/z for each compound of interest with its partial pressure in the perme- ate stream. Moreover, mass spectrometry proves to be a powerful tool for studying the fractionation of aromas, recovered by integrated pervaporation–condensation processes, allowing on-line monitoring of each solute vapour in the permeate stream. Additionally, this technique enables for accurate transient studies due to the possibility of acquiring one data point every 12–15s (potentially less, if desired) in real-time.

The isolation and characterization of a new metalloprotein containing Cu and Fe atoms is reported. The as-isolated Cu-Fe protein shows an UV-visible spectrum with absorption bands at 320 nm, 409 nm and 615 nm. Molecular mass of the native protein along with denaturating electrophoresis and mass spectrometry data show that this protein is a multimer consisting of 14 +/- 1 subunits of 15254.3 +/- 7.6 Da. Mossbauer spectroscopy data of the as-isolated Cu-Fe protein is consistent with the presence of [2Fe-2S](2+) centers. Data interpretation of the dithionite reduced protein suggest that the metallic cluster could be constituted by two ferromagnetically coupled [2Fe-2S](+) spin delocalized pairs. The biochemical properties of the Cu-Fe protein are similar to the recently reported molybdenum resistance associated protein from Desulfovibrio, D. alaskensis. Further-more, a BLAST search from the DNA deduced amino acid sequence shows that the Cu-Fe protein has homology with proteins annotated as zinc resistance associated proteins from Desulfovibrio, D. alaskensis, D. vulgaris Hildenborough, D. piger ATCC 29098. These facts suggest a possible role of the Cu-Fe protein in metal tolerance. (C) 2009 Published by Elsevier Inc.

A presente Dissertação consiste num estudo do comportamento competitivo de entidades, produtoras e comercializadoras de energia eléctrica, num período de tempo antes e após a entrada em vigor do Mercado Ibérico de Electricidade (MIBEL). Esse período de tempo engloba os meses entre Abril de 2007 e Abril de 2008, sendo que o MIBEL teve início a 1 de Julho de 2007. O estudo incide principalmente no grau de competitividade ou ausência da mesma, que as referidas entidades adoptaram após o começo do MIBEL. Para poder efectuar essa avaliação, recorre-se a uma abordagem baseada em variações conjecturais, que constituem um indicador desse grau de competitividade. Ou seja, as variações conjecturais representam a convicção que determinada empresa X tem face ao comportamento que outra empresa Y adoptará após uma acção efectuada pela empresa X. Por acção depreende-se, por exemplo, uma redução ou um aumento da quantidade de produção de uma empresa. Se o valor da variável conjectural for 0, as empresas encontram-se a concorrer à Cournot. Caso seja -1, as empresas encontram-se em concorrência perfeita. Por outro lado, se o valor da variação conjectural for superior a 0 pode existir uma situação de conluio entre as empresas. De modo a obter estes valores das variações conjecturais, utiliza-se e readapta-se um simulador de mercado diário, denominado SiMEC - Simulador do Mercado Eléctrico e do Carbono. Este simulador, que anteriormente simulava apenas períodos temporais precedentes ao MIBEL, foi reajustado de modo a incluir dados pós MIBEL. Finalmente, são então realizados gráficos comparativos e retiradas conclusões acerca do comportamento das entidades face a alterações na estrutura do mercado de electricidade.

Nitric Oxide Reductase (NOR) is an integral membrane protein performing the reduction of NO to N(2)O. NOR is composed of two subunits: the large one (NorB) is a bundle of 12 transmembrane helices (TMH). It contains a b type heme and a binuclear iron site, which is believed to be the catalytic site, comprising a heme b and a non-hemic iron. The small subunit (NorC) harbors a cytochrome c and is attached to the membrane through a unique TMH. With the aim to perform structural and functional studies of NOR, we have immunized dromedaries with NOR and produced several antibody fragments of the heavy chain (VHHs, also known as nanobodies). These fragments have been used to develop a faster NOR purification procedure, to proceed to crystallization assays and to analyze the electron transfer of electron donors. BIAcore experiments have revealed that up to three VHHs can bind concomitantly to NOR with affinities in the nanomolar range. This is the first example of the use of VHHs with an integral membrane protein. Our results indicate that VHHs are able to recognize with high affinity distinct epitopes on this class of proteins, and can be used as versatile and valuable tool for purification, functional study and crystallization of integral membrane proteins.

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