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NMR and molecular modelling studies on elastase inhibitor-peptides for wound ma...

Barros, Sandra Cerqueira; Louro, Ricardo O.; Micaelo, N. M.; Martins, José A.; Marcos, João Carlos; Paulo, Artur Cavaco

Proteases play an important and critical role in the physiological process of wound repair. However, excessive and unregulated release of proteolytic enzymes (e.g., elastase) mediates abnormal degradation of healthy tissues, which leads to inflammatory disorders such as chronic wounds. Thus, it is of therapeutic interest to develop novel synthetic inhibitor-peptides of elastase, which can restore the balance be...


Recovery and partitioning of fibrinolytic protease from Bacillus sp. UFPEDA 485...

Souza, F. A. S. D.; Nascimento, J. C. S.; Marques, D. A. V.; Marcos, João Carlos; Porto, T. S.; Teixeira, J. A.; Porto, A .L .F.

Fibrinolytic proteases produced by Bacillus sp. has attracted interest in the pharmaceutical industry as a promising alternative in thrombolytic therapy due to their effectiveness in degrading fibrin, its production requiring the development of an efficient recovery process. Aqueous two-phase systems (ATPS) have been recognized as an efficient and economical process for recovering enzymes. To optimize the recov...


Characterization of potential elastase inhibitor-peptides regulated by a molecu...

Barros, Sandra Cerqueira; Martins, J. A. R.; Marcos, João Carlos; Paulo, Artur Cavaco

Elastase plays an important role in wound healing process, degrading damaged tissue and allowing complete tissue recovery. The levels of human neutrophil elastase (HNE) are usually controlled by endogenous inhibitors. However, in the presence of high levels of elastase, like the ones present in chronic wounds, the inhibitors cannot overcome this overproduction and the enzyme starts to degrade the surrounding he...


Lysozyme and bovine serum albumin partitioning in polyethylene glycol–phenylala...

Silvério, Sara C.; Ferreira, Luísa A.; Martins, J. A.; Marcos, João Carlos; Macedo, E. A.; Teixeira, J. A.

Polyethylene glycol (PEG) with average molecular weight of 8000 was used as a model polymer and modified by introducing an amino acid (phenylalanine) in its hydroxyl terminals. The effect of different percentages of the conjugate polymer in relation to the total polymer on binodal curve of PEG-8000–potassium phosphate buffer (KPB), pH 7.2 aqueous two-phase systems and solute partitioning was explored. The parti...


Influence of secretory leukocyte protease inhibitor (SLPI) basedeptides on elas...

Barros, Sandra Cerqueira; Martins, J. A. R.; Marcos, João Carlos; Paulo, Artur Cavaco

Chronic nonhealing skin wounds, such as leg ulcers and pressure sores, represent a major clinical problem and a financial burden for the health care systems. Chronic wounds are characterized by prolonged inflammatory phase that results in high levels of elastase, reactive oxygen species (ROS), and diminished growth factor activity. Under normal physiological conditions, elastase is a powerful host defence and i...


Purification of plasmid DNA from clarified and non-clarified Escherichia coli l...

Nunes, C. Caramelo; Gabriel, M. F.; Almeida, Paulo; Marcos, João Carlos; Tomaz, C. T.

In this study, berenil was tested as a ligand, specifically to purify plasmids of different sizes pVAX1-LacZ (6.05 Kbp) and pCAMBIA-1303 (12.361 Kbp) from clarified Escherichia coli alkaline lysates. For this purpose, chromatographic experiments were performed using Sepharose derivatized with berenil. The results showed that both pDNA molecules are completely purified using lower amounts of salt in the eluent t...


Electrochemical generation of hydroxyl radicals for protein oxidation studies

Oliveira, Raquel; Bento, M. Fátima; Geraldo, M. Dulce; Marcos, João Carlos; Rodrigues, João

In the last decades, different studies shown that the increase in oxidative stress is associated with the risk factors implicated in the pathophysiology of many chronic diseases. Therefore it is important to evaluate the role that hydroxyl radicals-mediated damage may play in major human disorders. Proteins are important targets as they compose ca. 70% of cells. To understand the impact of specific modification...


Estudo dos danos oxidativos provocados em proteínas por radicais hidroxilo gera...

Santos, Carlos; Pereira, Nelson; Oliveira, Raquel; Bento, M. Fátima; Geraldo, M. Dulce; Marcos, João Carlos

Os radicais HO são espécies altamente reativas usualmente designadas de espécies reativas de oxigénio (ROS). Estas espécies podem provocar danos importantes em organismos vivos, estando na origem do stresse oxidativo1, 2. As proteínas, dada a sua abundância em células, constituem alvos importantes do ataque por ROS, sendo assim relevante caracterizar as suas modificações oxidativas. Neste contexto é importante ...


Specific berenil–DNA interactions : an approach for separation of plasmid isofo...

Caramelo-Nunes, C.; Tente, T.; Almeida, P.; Marcos, João Carlos; Tomaz, C. T.

Small molecules, like some antibiotics and anticancer agents that bind DNA with high specificity can represent a relevant alternative as ligands in affinity processes for plasmid DNA (pDNA) purification. In the present study, pDNA binding affinities of berberine, berenil, kanamycin and neomycin were evaluated by a competitive displacement assay with ethidium bromide using a fluorimetric titration technique. The...


Influence of the chemical structure of the polymer in the partition of proteins...

Silvério, Sara C.; Madeira, Pedro; Martins, J. A. R.; Marcos, João Carlos; Teixeira, J. A.; Macedo, Eugénia


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    Financiadores do RCAAP

Fundação para a Ciência e a Tecnologia Universidade do Minho   Governo Português Ministério da Educação e Ciência Programa Operacional da Sociedade do Conhecimento União Europeia