Environmental assessment of proton exchange membrane fuel cell platinum catalyst recycling

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Protection by polyphenol extract from olive stones against apoptosis produced by oxidative stress in human neuroblastoma cells

Objective: We evaluated the protective activity of an extract from a by-product such as olive stones, through its ability to inhibit H2O2 induced apoptosis in the SH-SY5Y human neuroblastoma cell line. Material and methods: To such end, 20,000 cells/well were cultivated and differentiation with retinoic acid was initiated. Once the cells were differentiated, apoptosis was induced with and without H2O2 extract. Finally, cDNA extraction was performed, and pro-apoptotic genes Bax and anti-apoptotic genes Bcl-2 were analyzed. Quantification of the gene expression was performed using the GAPDH gene marker. Results: Cell viability with the extract is 97.6% (SD 5.7) with 10 mg/l and 62.8% (SD 1.2) to 50 mg/l, using 10 mg/l for the biomarker assay. The retinoic acid differentiated SH-S cell line (10 µM) shows a clear apoptosis when treated with H2O2 150 µM, with a Bax/Bcl-2 ratio of 3.75 (SD 0.80) in contrast to the differentiated control cells subjected to H2O2 and with extract, which have the same ratio of 1.02 (SD 0.01-0.03). Conclusion: The olive stone extract shows anti-apoptotic activity in the provoked cell death of SH-SY5Y human neuroblastoma cells in their normal state, defending them from oxidative stress which produces a significant increase in the apoptotic gene ratio in contrast to anti-apoptotic genes (Bax/Bcl-2).

Evaluation of the antioxidant activity and antitumor activity of marine invertebrates extracts

Dissertação de Mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologias, Universidade do Algarve, 2014

Estabilidade térmica dos compostos biologicamente ativos do colostro

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Síntese enzimática de ésteres de açúcar: surfactantes e polímeros como novos materiais ambientalmente seguros

Sugar esters are substances which possess surfactant, antifungical and bactericidal actions and can be obtained through two renewable sources of raw materials: sugars and vegetable oils. Their excellent biodegradability, allied to lhe fact that they are non toxic, insipid, inodorous, biocompatible, no-ionic, digestible and because they can resist to adverse conditions of temperature, pH and salinity, explain lhe crescent use of these substances in several sections of lhe industry. The objective of this thesis was to synthesize and characterize surfactants and polymers containing sugar branched in their structures, through enzymatic transesterification of vinyl esters and sugars, using alkaline protease from Bacillus subtilis as catalyst, in organic medium (DMF).Three types of sugars were used: L-arabinose, D-glucose and sucrose and two types of vinyl esters: vinyl laurate and vinyl adipate. Aiming to reach high conversions from substrates to products for a possible future large scale industrial production, a serie of variables was optimized, through Design of Experiments (DOE), using Response Surface Methodology (RSM).The investigated variables were: (1) enzyme concentration; (2) molar reason of substrates; (3) water/solvent rale; (4) temperature and (5) time. We obtained six distinct sugar esters: 5-0-lauroyl L-arabinose, 6-0-lauroyl D-glucose, 1'-O-lauroyl sucrose, 5-0-vinyladipoyl L-arabinose, 6-0-vinyladipoyl D-glucose and 1 '-O-vinyladipoyl sucrose, being lhe last three polymerizable. The progress of lhe reaction was monitored by HPLC analysis, through lhe decrease of sugar concentration in comparison to lhe blank. Qualitative analysis by TLC confirmed lhe formation of lhe products. In lhe purification step, two methodologies were adopted: (1) chromatographic column and (2) extraction with hot acetone. The acylation position and lhe chemical structure were determined by 13C-RMN. The polymerization of lhe three vinyl sugar esters was possible, through chemical catalysis, using H2O2 and K2S2O8 as initiators, at 60°C, for 24 hours. IR spectra of lhe monomers and respective polymers were compared revealing lhe disappearance of lhe vinyl group in lhe polymer spectra. The molar weights of lhe polymers were determined by GPC and presented lhe following results: poly (5-0-vinyladipoyl L-arabinose): Mw = 7.2 X 104; PD = 2.48; poly (6-0-vinyladipoyl D-glucose): Mw = 2.7 X 103; PD = 1.75 and poly (1'-O-vinyladipoyl sucrose): Mw = 4.2 X 104; PD = 6.57. The six sugar esters were submitted to superficial tension tests for determination of the critical micelle concentrations (CMC), which varied from 122 to 167 ppm. Finally, a study of applicability of these sugar esters, as lubricants for completion fluids of petroleum wells was' accomplished through comparative analysis of lhe efficiency of these sugar esters, in relation to three commercial lubricants. The products synthesized in this thesis presented equivalent or superior action to lhe tested commercial products

Avaliação de metodologia de preparo de amostra de petróleo por digestão ácida auxiliada por microondas para determinação de metais por ICP-OES

This project describes a methodology optimization that would allow for a more efficient microwave assisted digestion process for petroleum samples. With the possible chance to vary various factors at once to see if any one factor was significant enough in the answers, experimental planning was used. Microwave assisted digestion allows, through the application of potency, an increasing number of collisions between the HNO3 and H2O2 molecules, favoring sample opening for complex matrixes. For this, a 24 factorial experimental planning was used, varying potency, time and the volumes for HNO3 65% and H2O2 30%. To achieve the desired answers, several elements were monitored (C, Cu, Cr, Fe, Ni, Zn and V) through Inductively coupled plasma atomic emission spectroscopy (ICP-OES). With this initial study it was noticed that the HNO3 was not a significant factor for any of the statistical studies for any of the analytes and the other 3 factors and their interactions showed statistical significance. A Box Behnken experimental planning was used taking in consideration 3 factors: H2O2 volume, time (min) and Potency (W), Nitric Acid kept at 4mL for a mass of 0,1g of petroleum. The results were extremely satisfying showing higher efficiency in the digestion process and taking in a responsibility between the answers for each analyte and the carbon monitoring was achieved in the following conditions: 7mL of H2O2, 700 Watts of potency and a reaction time of 7 minutes with 4mL de HNO3 for a mass of 0,1g of petroleum. The optimized digestion process was applied to four different petroleum samples and the analytes determined by ICP-OES

Functionalized Carbon Micro/Nanostructures for Biomolecular Detection

Advancements in the micro-and nano-scale fabrication techniques have opened up new avenues for the development of portable, scalable and easier-to-use biosensors. Over the last few years, electrodes made of carbon have been widely used as sensing units in biosensors due to their attractive physiochemical properties. The aim of this research is to investigate different strategies to develop functionalized high surface carbon micro/nano-structures for electrochemical and biosensing devices. High aspect ratio three-dimensional carbon microarrays were fabricated via carbon microelectromechanical systems (C-MEMS) technique, which is based on pyrolyzing pre-patterned organic photoresist polymers. To further increase the surface area of the carbon microstructures, surface porosity was introduced by two strategies, i.e. (i) using F127 as porogen and (ii) oxygen reactive ion etch (RIE) treatment. Electrochemical characterization showed that porous carbon thin film electrodes prepared by using F127 as porogen had an effective surface area (Aeff 185%) compared to the conventional carbon electrode. To achieve enhanced electrochemical sensitivity for C-MEMS based functional devices, graphene was conformally coated onto high aspect ratio three-dimensional (3D) carbon micropillar arrays using electrostatic spray deposition (ESD) technique. The amperometric response of graphene/carbon micropillar electrode arrays exhibited higher electrochemical activity, improved charge transfer and a linear response towards H2O2 detection between 250μM to 5.5mM. Furthermore, carbon structures with dimensions from 50 nano-to micrometer level have been fabricated by pyrolyzing photo-nanoimprint lithography patterned organic resist polymer. Microstructure, elemental composition and resistivity characterization of the carbon nanostructures produced by this process were very similar to conventional photoresist derived carbon. Surface functionalization of the carbon nanostructures was performed using direct amination technique. Considering the need for requisite functional groups to covalently attach bioreceptors on the carbon surface for biomolecule detection, different oxidation techniques were compared to study the types of carbon–oxygen groups formed on the surface and their percentages with respect to different oxidation pretreatment times. Finally, a label-free detection strategy using signaling aptamer/protein binding complex for platelet-derived growth factor oncoprotein detection on functionalized three-dimensional carbon microarrays platform was demonstrated. The sensor showed near linear relationship between the relative fluorescence difference and protein concentration even in the sub-nanomolar range with an excellent detection limit of 5 pmol.

Nuclear Magnetic Resonance Spectroscopic and Computational Investigations of Chloroperoxidase Catalyzed Regio- and Enantio-Selective Transformations

Chloroperoxidase (CPO) is the most versatile heme-containing enzyme that catalyzes a broad spectrum of reactions. The remarkable feature of this enzyme is the high regio- and enantio-selectivity exhibited in CPO-catalyzed oxidation reactions. The aim of this dissertation is to elucidate the structural basis for regio- and enantio-selective transformations and investigate the application of CPO in biodegradation of synthetic dyes. To unravel the mechanism of CPO-catalyzed regioselective oxidation of indole, the dissertation explored the structure of CPO-indole complex using paramagnetic relaxation and molecular modeling. The distances between the protons of indole and the heme iron revealed that the pyrrole ring of indole is oriented toward the heme with its 2-H pointing directly at the heme iron. This provides the first experimental and theoretical explanation for the "unexpected" regioselectivity of CPO-catalyzed indole oxidation. Furthermore, the residues including Leu 70, Phe 103, Ile 179, Val 182, Glu 183, and Phe 186 were found essential to the substrate binding to CPO. These results will serve as a lighthouse in guiding the design of CPO mutants with tailor-made activities for biotechnological applications. To understand the origin of the enantioselectivity of CPO-catalyzed oxidation reactions, the interactions of CPO with substrates such as 2-(methylthio)thiophene were investigated by nuclear magnetic resonance spectroscopy (NMR) and computational techniques. In particular, the enantioselectivity is partly explained by the binding orientation of substrates. In third facet of this dissertation, a green and efficient system for degradation of synthetic dyes was developed. Several commercial dyes such as orange G were tested in the CPO-H2O2-Cl- system, where degradation of these dyes was found very efficient. The presence of halide ions and acidic pH were found necessary to the decomposition of dyes. Significantly, the results revealed that this degradation of azo dyes involves a ferric hypochlorite intermediate of CPO (Fe-OCl), compound X.

Niobic acid nanoparticle catalysts for the aqueous phase transformation of glucose and fructose to 5-hydroxymethylfurfural

A family of bulk and SBA-15 supported peroxo niobic acid sols were prepared by peptisation of niobic acid precipitates with H2O2 as heterogeneous catalysts for aqueous phase glucose and fructose conversion to 5-hydroxymethylfurfural (5-HMF). Niobic acid nanoparticles possess a high density of Brønsted and Lewis acid sites, conferring good activity towards glucose and fructose conversion, albeit with modest 5-HMF yields under mild reaction conditions (100 °C). Thermally-induced niobia crystallisation suppresses solid acidity and activity. Nanoparticulate niobic acid dispersed over SBA-15 exhibits pure Brønsted acidity and an enhanced Turnover Frequency for fructose dehydration.

Fenton-like degradation of Bisphenol A catalyzed by mesoporous Cu/TUD-1

A family of copper oxide catalysts with loadings spanning 1–5 wt% were dispersed on a three dimensional, mesoporous TUD-1 silica through a hydrothermal, surfactant-free route employing tetraethylene glycol as a structure-directing agent. Their bulk and surface properties were characterized by N2 physisorption, XRD, DRUVS, EPR, TEM and Raman spectroscopy, confirming the expected mesoporous wormhole/foam support morphology and presence of well-dispersed CuO nanoparticles (∼5–20 nm). The catalytic performance of Cu/TUD-1 was evaluated as heterogeneous Fenton-like catalysts for Bisphenol A (BPA) oxidative degradation in the presence of H2O2 as a function of [H2O2], and CuO loading. Up to 90.4% of 100 ppm BPA removal was achieved over 2.5 wt% Cu/TUD-1 within 180 min, with negligible Cu leaching into the treated water.

A non-classical LysR-type transcriptional regulator PA2206 is required for an effective oxidative stress response in Pseudomonas aeruginosa

LysR-type transcriptional regulators (LTTRs) are emerging as key circuit components in regulating microbial stress responses and are implicated in modulating oxidative stress in the human opportunistic pathogen Pseudomonas aeruginosa. The oxidative stress response encapsulates several strategies to overcome the deleterious effects of reactive oxygen species. However, many of the regulatory components and associated molecular mechanisms underpinning this key adaptive response remain to be characterised. Comparative analysis of publically available transcriptomic datasets led to the identification of a novel LTTR, PA2206, whose expression was altered in response to a range of host signals in addition to oxidative stress. PA2206 was found to be required for tolerance to H2O2 in vitro and lethality in vivo in the Zebrafish embryo model of infection. Transcriptomic analysis in the presence of H2O2 showed that PA2206 altered the expression of 58 genes, including a large repertoire of oxidative stress and iron responsive genes, independent of the master regulator of oxidative stress, OxyR. Contrary to the classic mechanism of LysR regulation, PA2206 did not autoregulate its own expression and did not influence expression of adjacent or divergently transcribed genes. The PA2214-15 operon was identified as a direct target of PA2206 with truncated promoter fragments revealing binding to the 5'-ATTGCCTGGGGTTAT-3' LysR box adjacent to the predicted -35 region. PA2206 also interacted with the pvdS promoter suggesting a global dimension to the PA2206 regulon, and suggests PA2206 is an important regulatory component of P. aeruginosa adaptation during oxidative stress.

Branqueamento dentário

A estética dentária tem recebido bastante enfoque nos últimos anos, particularmente devido à importância a que a população atribui à aparência estética do sorriso. É, assim, desejado um sorriso o mais branco possível e que de preferência seja fácil de obter, eficaz, rápido, económico e que seja o menos invasivo possível. No entanto, muitos pacientes apresentam frequentemente dentes com cor alterada, comprometendo desta forma a estética do sorriso. O branqueamento dentário é uma técnica não invasiva, conservadora que não altera a forma natural do dente, e que permite alterações estéticas consideráveis. A procura de uma melhoria estética a todos os níveis, leva a que o Branqueamento dentário se apresente hoje como método de eleição para a remoção da pigmentação dentária. Recorre-se assim a substâncias oxidantes, que na maioria dos casos têm origem no Peróxido de Hidrogénio (H2O2). Um dos efeitos secundários ao Branqueamento, prende-se com a sensibilidade dentária, sendo que esta pode originar algum desconforto ou mesmo ser condicionante para a não realização ou término do tratamento. Para se atingir sucesso num tratamento branqueador é da maior importância o diagnóstico preciso da etiologia da alteração de cor, por isso uma anamnese detalhada e um exame clínico e dentário são da maior importância para se poder aconselhar o paciente pelo melhor tratamento a adotar. O objetivo deste trabalho foi avaliar a informação científica disponível sobre as técnicas disponíveis para realizar branqueamento dentário, vantagens e desvantagens de cada técnica, agentes branqueadores utilizados, mecanismos de ação e os seus efeitos adversos. Para tal foi efetuada uma pesquisa nas bases de dados PubMed e B-On de artigos publicados entre 2006-2016 com as seguintes palavras-chave: dental bleaching, teeth whitening, peroxides, branqueamento dentário, clareamento dentário. O branqueamento dentário, apresenta algumas limitações e contra-indicações, assim como vários efeitos adversos, que devem ser do conhecimento do Médico para este poder intervir devidamente. Foi percetível que um tratamento branqueador depende de inúmeros fatores e que a forma de atuação do profissional é tão importante para o sucesso do tratamento como o tipo de agente branqueador utilizado.

Efficient removal of paracetamol using LaCu1-xMxO3 (M=Mn, Ti) perovskites as heterogeneous Fenton-like catalysts

A series of perovskite-like oxides LaCu1-xMxO3 (M=Mn, Ti; 0.0 ⩽ x ⩽ 0.8) was prepared by amorphous citrate decomposition and characterized by XRD, ICP-OES and XPS techniques. The catalysts were tested in the Fenton-like degradation of paracetamol with H2O2, under mild reaction conditions, 25 °C and nearly neutral pH. Values of decomposition of paracetamol between 80 and 97% at 300 min were achieved for most of samples. The presence of the Cu2+/Cu+ pair at the surface of the catalysts is necessary to carry out the reaction and the catalysts containing higher amount of copper at the surface, resulted to be more active. The leaching of metals was less than 1%, which discards the contribution of the homogenous Fenton-like reaction and remarks the high stability of the metals into the mixed oxide network. The catalytic activity of LaCu0.8Mn0.2O3 was maintained after three cycles of reaction, which proves the stability and reusability of the catalyst.

he genome and genetics of a high oxidative stress tolerant Serratia sp. LCN16 isolated from the plant parasitic nematode Bursaphelenchus xylophilus

Background: Pine wilt disease (PWD) is a worldwide threat to pine forests, and is caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus. Bacteria are known to be associated with PWN and may have an important role in PWD. Serratia sp. LCN16 is a PWN-associated bacterium, highly resistant to oxidative stress in vitro, and which beneficially contributes to the PWN survival under these conditions. Oxidative stress is generated as a part of the basal defense mechanism used by plants to combat pathogenic invasion. Here, we studied the biology of Serratia sp. LCN16 through genome analyses, and further investigated, using reverse genetics, the role of two genes directly involved in the neutralization of H2O2, namely the H2O2 transcriptional factor oxyR; and the H2O2-targeting enzyme, catalase katA. Results: Serratia sp. LCN16 is phylogenetically most closely related to the phytosphere group of Serratia, which includes S. proteamaculans, S. grimessi and S. liquefaciens. Likewise, Serratia sp. LCN16 shares many features with endophytes (plant-associated bacteria), such as genes coding for plant polymer degrading enzymes, iron uptake/ transport, siderophore and phytohormone synthesis, aromatic compound degradation and detoxification enzymes. OxyR and KatA are directly involved in the high tolerance to H2O2 of Serratia sp. LCN16. Under oxidative stress, Serratia sp. LCN16 expresses katA independently of OxyR in contrast with katG which is under positive regulation of OxyR. Serratia sp. LCN16 mutants for oxyR (oxyR::int(614)) and katA (katA::int(808)) were sensitive to H2O2 in relation with wild-type, and both failed to protect the PWN from H2O2-stress exposure. Moreover, both mutants showed different phenotypes in terms of biofilm production and swimming/swarming behaviors. Conclusions: This study provides new insights into the biology of PWN-associated bacteria Serratia sp. LCN16 and its extreme resistance to oxidative stress conditions, encouraging further research on the potential role of this bacterium in interaction with PWN in planta environment.

Caracterização analítica de rochas silicatadas e avaliação de seu potencial agrícola como fonte de potássio.

O presente estudo avaliou a eficiência agronômica da aplicação de pó de rochas silicatadas como fonte de potássio em cultivos consecutivos de milheto e alfafa em casa de vegetação. As rochas Biotita Xisto (BX), Sienito Ceraíma (TA15), Mafurito Acreúna (TA21), Sienito SANW (TA20), Biotita Gnaísse (EL02) e Fonolito Curimbaba (FN) foram aplicadas como fontes de potássio de liberação lenta em três doses de 1,25; 2,50 e 5,0 g de rocha moída por kg de solo. Cloreto de potássio foi utilizado como fertilizante convencional em doses de 0,10; 0,20 e 0,40 g de KCl kg-1 de solo. O solo utilizado nos experimentos foi um latossolo amarelo distrófico de textura média, com teor de potássio de 1,2 mmolc dm-3 solo. A rocha TA21 foi testada sem e com correção da acidez do solo. Com a finalidade de observar a dissolução das rochas em um ambiente com plantas foram realizados cinco cultivos consecutivos: (1) milheto (62 dias de cultivo); (2), (3) e (4) alfafa (80, 115 e 155 dias de cultivo, respectivamente) e (5) milheto, com 80 dias de cultivo. A produção de matéria seca no cultivo da alfafa foi significativamente menor do que o cultivo do milheto. As amostras de forrageira foram digeridas e quantificadas empregando o método de decomposição assistida por radiação micro-ondas com HNO3 + H2O2 para a determinação do teor de potássio extraído pelas plantas. As amostras de rochas foram caracterizadas por fluorescência de raios X, difração de raios X, microscopia eletrônica de varredura e distribuição de tamanho das partículas e submetidas a ensaios com solução extratora Mehlich-1, ácido acético e ácido fumárico 0,05mol L-1, em diferentes tempos de extração. Todos os extratos foram quantificados por espectrometria de emissão óptica com plasma acoplado indutivamente (ICP OES), sendo determinados os teores de Al, Ca, K, Mg, Mn, Na, P e Zn. O método da resina trocadora de íons foi aplicado para todas as amostras de rochas avaliadas, sendo quantificados K, Ca, Mg e P. Potássio solúvel foi determinado por fotometria de chama após extração com água quente, método oficial do Ministério da Agricultura, Pecuária e Abastecimento (MAPA). Para extração drástica foram avaliados dois métodos de decomposição assistida por radiação microondas: com uso de água régia e água régia invertida com adição de H2O2 e 2 etapas de pré-decomposição. Os elementos Al, Ca, K, Mg, Mn, Na e Zn foram quantificados por ICP OES e os resultados obtidos foram comparados por meio do teste t pareado, que indicou não haver diferença significativa entre os métodos de decomposição no intervalo de 95% de confiança. A partir das concentrações obtidas nas extrações e da quantidade de potássio extraída nos cultivos foi realizado o cálculo de correlação entre os extratores e as plantas, considerando a eficiência agronômica. O uso da calagem nos solos que receberam adição da rocha TA21 influenciou significativamente a absorção de potássio, sendo que foi observado efeito relativo à liberação de K apenas quando essa rocha foi adicionada ao solo sem tratamento prévio com a calagem. A ordem decrescente de eficiência de extração de potássio, considerando o total aplicado nos tratamentos e o K extraído pelas plantas foi EL02 > TA21-SC > KCl > FN> BX> TA20 > TA15 > TA21. Os dois métodos de decomposição desenvolvidos apresentaram melhor correlação com o potássio extraído das rochas EL02. O estudo de casa de vegetação corroborou com os ensaios de extração realizados em laboratório, que indicaram que a rocha EL02 é a que possui o maior percentual de liberação de K em relação às demais rochas avaliadas.