Percorrer por autor "Coelho, Andreia"
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- Cry1A(b)16 toxin from Bacillus thuringiensis: Theoretical refinement of three-dimensional structure and prediction of peptides as molecular markers for detection of genetically modified organismsPublication . Plácido, Alexandra; Coelho, Andreia; Abreu Nascimento, Lucas; Gomes Vasconcelos, Andreanne; Barroso, M. Fátima; Ramos-Jesus, Joilson; Costa, Vladimir; Lima, Francisco das Chagas Alves; Delerue-Matos, Cristina; Ramos, Ricardo Martins; Marani, Mariela M.; Leite, José Roberto de Souza de AlmeidaTransgenic maize produced by the insertion of the Cry transgene into its genome became the second most cultivated crop worldwide. Cry gene from Bacillus thuringiensis kurstaki expresses protein derivatives of crystalline endotoxins which confer insect resistance onto the maize crop. Mandatory labeling of processed food containing or made by genetically modified organisms is in force in many countries, so, it is very urgent to develop fast and practical methods for GMO identification, for example, biosensors. In the absence of an available empirical structure of Cry1A(b)16 protein, a theoretical model was effectively generated, in this work, by homology modeling and molecular dynamics simulations based on two available homologous protein structures. Molecular dynamics simulations were carried out to refine the selected model, and an analysis of its global structure was performed. The refined models of Cry1A(b)16 showed a standard fold and structural characteristics similar to those seen in Bacillus thuringiensis Cry1A(a) insecticidal toxin and Bacillus thuringiensis serovar kurstaki Cry1A(c) toxin. After in silico analysis of Cry1A(b)16, two immunoreactive candidate peptides were selected and specific polyclonal antibodies were produced resulting in antibody-peptide interaction. Biosensing devices are expected to be developed for detection of the Cry1A(b) protein as a marker of transgenic maize in food. Proteins 2017; 85:1248-1257. © 2017 Wiley Periodicals, Inc.
- DNA-based sensor against nitrite oxide radical: Evaluation of total antioxidant capacity in beveragesPublication . Cruz, Diana; Barroso, M. Fátima; Ramalhosa, Maria João; Coelho, Andreia; Silva, Hélder da; Duarte, Abel J.; González-García, Maria Begoña; Carvalho, Ana P.; Delerue-Matos, CristinaReactive nitrogen and oxygen species are produced in cells and play an important role in the oxidative metabolism; when in abnormal concentrations, they are able to induce oxidative damage on biomolecules, namely in DNA. In this paper it is described, an electrochemical DNA-based sensor against NOradical dot radical developed for total antioxidant capacity (TAC) evaluation. The sensor consisted on dA20 (adenine-rich oligonucleotide) physically adsorbed into carbon paste electrode (CPE). When this dA20-CPE was damaged, by immersion in a freshly generated NOradical dot radical, a protective effect onto dA20 was observed in the presence of antioxidants (ascorbic, gallic, caffeic, p-coumaric acids). Electrochemical studies were performed through square wave voltammetry. The construction of the sensor is simple, fast and the results indicated that the DNA-based sensor is suitable, accurate, and can be used to the assessment of TAC in commercial samples of juices.
- Eruca sativa: Benefits as antioxidants source versus risks of already banned pesticidesPublication . Maia, M. Luz; Correia-Sá, Luísa; Coelho, Andreia; Barroso, M. Fátima; Domingues, Valentina F.; Delerue-Matos, CristinaEruca sativa (rocket salad) has been intensely consumed all over the world, insomuch as, this work was undertaken to evaluate the antioxidant status and the environmental contamination (positive and negative nutritional contribution) of leaves and stems from this vegetable. Antioxidant capacity of rocket salad was assessed by mean of optical methods, such as the total phenolic content (TPC), reducing power assay and DPPH radical scavenging activity. The extent of the environmental contamination was reached through the quantification of thirteen organochlorine pesticides (OCP) by using gas chromatography coupled with electron-capture detector (GC-ECD) and compound confirmations employing gas chromatography tandem mass-spectrometry (GC-MS/MS). The OCP residues were extracted by using Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) methodology.The extent of the environmental contamination was reached through the quantification of thirteen OCP by using gas chromatography coupled with electron-capture detector (GC-ECD) and compound confirmations employing GC-MS/MS. The OCP residues were extracted by using Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) methodology. This demonstrated that leaves presented more antioxidant activity than stems, emphasizing that leaves contained six times more polyphenolic compounds than stems. In what concerns the OCP occurrence, the average recoveries obtained at the three levels tested (40, 60 and 80 µg kg−1) ranged from 55% to 149% with a relative standard deviation of 11%, (except hexachrorobenzene). Three vegetables samples were collected from supermarkets and analysed following this study. According to data, only one sample achieved 16.21 of β-hexachlorocyclohexane, confirmed by GC-MS/MS. About OCP quantification, the data indicated that only one sample achieved 16.21 µg kg−1 of β-hexachlorocyclohexane, confirmed by GC-MS/MS, being the QuEChERS a good choice for the of OCPs extraction. Furthermore, the leaves consumption guaranty higher levels of antioxidants than stems.