bio

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