Ph.D. in Pharmaceutical and Clinical Biochemistry (FCUL, Portugal) Lic. in Biochemistry (FCUL, Portugal)
My research activity has been focused on: • Reactive oxygen and nitrogen species (ROS/RNS) metabolism – Mammalian formation/consumption of ROS/RNS, nitric oxide radical, superoxide anion radical and peroxides (hydrogen and lipidics) – Mechanisms of ROS/RNS-mediated diseases, namely in the pathologies associated with vascular dysfunction and alcohol hepatotoxicity • Human signalling pathways – Novel pathways of formation/consumption of signalling molecules, namely nitric oxide – In vitro studies with purified rat liver and human liver xanthine oxidase, xanthine dehydrogenase and aldehyde oxidase – In situ studies with human cell lines • New catalytic activities of molybdoenzymes – NADH oxidation, nitrite reduction, nitric oxide formation, carbon dioxide reduction (carbon dioxide scavenging) – Mechanistic and kinetic studies – Mammalian and bacterial enzymes • Structure-activity relationships in metalloenzymes – Mechanistic and kinetic studies – Small proteins mimetics (molybdenum-substituted rubredoxin) – Mammalian and bacterial enzymes (rat and human liver xanthine oxidase, xanthine dehydrogenase and aldehyde oxidase, bovine xanthine oxidase, Desulfovibrio aldehyde oxidoreductases and formate dehydrogenases, Marinobacter nitric oxide reductase)
Methodologies used: • Optimization of enzyme purification procedures – Chromatography (ionic exchange, adsorption, filtration), centrifugation, precipitation, electrophoresis • Kinetic characterisation – Reduction/oxidation of nitric oxide precursors, nitric oxide, metabolites relevant to ischemia injury, alcohol hepatotoxicity (purinic and pyridinic compounds), aldehydes metabolism (aromatic and aliphatic), carbon dioxide metabolism – Spectroscopic (UV-visible, fluorescence, EPR, NMR), HPLC and polarographic methodologies • Mechanistic studies – Enzymes redox active centres characterisation, electron transfer within the centres and substrates/products/inhibitors, aiming to characterise the reactions mechanism with atomic detail – EPR, NMR, mass, UV-visible spectroscopies, with substrates, products, specific inhibitors, isotopically labelled compounds and spin-traps • Quantification of enzymatic ROS/RNS formation (•NO, O2•-, H2O2, HO•, ONOO-), endogenous antioxidants (vitamins E and C, GSH, SOD, catalase), and oxidative modifications on proteins, nucleic acids and lipids – Spectrophotometric, fluorimetric, EPR, MS, HPLC, GC and electrophoretic methodologies, lipid purification techniques • Tissue cultures (human cell lines) – Fluorescence methodologies (probes to follow signalling molecules) • Models of oxidative stress
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