Percorrer por autor "Vaz, Daniela C."
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- Alginate films encapsulating polyphenols for nutraceutical and biomedical applicationsPublication . Lagoa, Ricardo; Vanat, Pavlo; Ferreira, Stefanie; Silva, João; Marques-da-Silva, Dorinda; Vaz, Daniela C.; Barreiros, Fátima; Rodrigues, Joaquim Rui; Santos Ribeiro, Vânia
- BSA-PEG Hydrogel: A Novel Protein-Ligand Binding 3D MatrixPublication . Coelho, Carlos D. F.; Jesus, João A.; Vaz, Daniela C.; Lagoa, Ricardo; Moreno, Maria JoãoHydrogel materials have good biomimetic properties and high potential for biomedical and bioanalytical applications. In this work, a hydrogel of serum albumin crosslinked with poly-(ethylene glycol) was prepared and characterized for its water content, protein structure and stability. The ability of the hydrogel to bind small molecule ligands with different hydrophobicity was evaluated using a homologous series of amphiphiles (NBD-Cn, n = 4, 6 and 8) and the calculated binding affinities were similar to that of free protein in solution. Overall, the results indicate this type of hydrogel system as a convenient tool for studying the binding of xenobiotics to tissue proteins.
- Chemical characterization of fungal chitosan for industrial applicationsPublication . Cruz, Pedro; Vaz, Daniela C.; Ruben, Rui B.; Avelelas, Francisco; Silva, Susana; Campos, Maria
- Corema album spp: Edible wild crowberries with a high content in minerals and organic acidsPublication . Brito, Catarina; Bertotti, Taciana; Primitivo, Maria João; Neves, Marta; Pires, Cristiana L.; Cruz, Pedro F.; Martins, Patrícia A. T.; Rodrigues, Ana Cristina; Moreno, Maria João; Brito, Rui M. M.; Campos, Maria; Vaz, Daniela C.; Pessoa, Maria Fernanda; Lidon, Fernando; Reboredo, Fernando; Santos Ribeiro, VâniaEdible wild plants are part of the ethnobotanical and gastronomic heritage of different geographical areas. Corema album (L.) D. Don is an endemic species of the dune systems of the Atlantic coast of the Iberian Peninsula. The aerial parts of Corema album are a source of nutrients and antioxidants. The Corema album white berry (Portuguese crowberry) is rich in calcium, iron, and zinc. The plant also shows high phenolic content and antioxidant capacity associated with the leaves, fruit, and flowers. The presence of organic acids, namely phenolic acids, such as hydroxycinnamic acids, and long chain polyunsaturated fatty acids (PUFAs) omega-3 and omega-6 has also been confirmed. Toxicity studies evaluated by cell viability tests with human intestinal epithelium model cells (Caco-2) have shown that, at low concentrations, plant extracts may present beneficial effects.
- Development of novel alginate based hydrogel films for wound healing applicationsPublication . Pereira, Rúben; Carvalho, Anabela; Vaz, Daniela C.; Gil, M. H.; Mendes, Ausenda; Bártolo, PauloAlginate and Aloe vera are natural materials widely investigated and used in the biomedical field. In this research work, thin hydrogel films composed by alginate and Aloe vera gel in different proportions (95:5, 85:15 and 75:25, v/v) were prepared and characterized. The films were evaluated regarding the light transmission behavior, contact angle measurements, and chemical, thermal and mechanical properties. These thin hydrogel films, prepared by crosslinking reaction using 5% calcium chloride solution, were also investigated relatively to their water solubility and swelling behavior. Results showed that Aloe vera improved the transparency of the films, as well their thermal stability. The developed films present adequate mechanical properties for skin applications, while the solubility studies demonstrated the insolubility of the films after 24 h of immersion in distilled water. The water absorption and swelling behavior of these films were greatly improved by the increase in Aloe vera proportion.
- DIBMA nanodiscs keep α-synuclein foldedPublication . Adão, Regina; Cruz, Pedro F.; Vaz, Daniela C.; Fonseca, Fátima; Pedersen, Jannik Nedergaard; Ferreira-da-Silva, Frederico; Brito, Rui M. M.; Ramos, Carlos H. I.; Otzen, Daniel; Keller, Sandro; Bastos, Margaridaα-Synuclein (αsyn) is a cytosolic intrinsically disordered protein (IDP) known to fold into an α-helical structure when binding to membrane lipids, decreasing protein aggregation. Model membrane enable elucidation of factors critically affecting protein folding/aggregation, mostly using either small unilamellar vesicles (SUVs) or nanodiscs surrounded by membrane scaffold proteins (MSPs). Yet SUVs are mechanically strained, while MSP nanodiscs are expensive. To test the impact of lipid particle size on α-syn structuring, while overcoming the limitations associated with the lipid particles used so far, we compared the effects of large unilamellar vesicles (LUVs) and lipid-bilayer nanodiscs encapsulated by diisobutylene/maleic acid copolymer (DIBMA) on αsyn secondary-structure formation, using human-, elephant- and whale -αsyn. Our results confirm that negatively charged lipids induce αsyn folding in h-αsyn and e-αsyn but not in w-αsyn. When a mixture of zwitterionic and negatively charged lipids was used, no increase in the secondary structure was detected at 45 °C. Further, our results show that DIBMA/lipid particles (DIBMALPs) are highly suitable nanoscale membrane mimics for studying αsyn secondary-structure formation and aggregation, as folding was essentially independent of the lipid/protein ratio, in contrast with what we observed for LUVs having the same lipid compositions. This study reveals a new and promising application of polymer-encapsulated lipid-bilayer nanodiscs, due to their excellent efficiency in structuring disordered proteins such as αsyn into nontoxic α-helical structures. This will contribute to the unravelling and modelling aspects concerning protein-lipid interactions and α-helix formation by αsyn, paramount to the proposal of new methods to avoid protein aggregation and disease.
- Edible flowers of Helichrysum italicum: Composition, nutritive value, and bioactivitiesPublication . Primitivo, Maria João; Neves, Marta; Pires, Cristiana L.; Cruz, Pedro F.; Brito, Catarina; Rodrigues, Ana C.; Carvalho, Carla C. C. R. de; Mortimer, Megan M.; Moreno, Maria João; Brito, Rui M. M.; Taylor, Edward J.; Millson, Stefan H.; Reboredo, Fernando; Campos, Maria; Vaz, Daniela C.; Santos Ribeiro, VâniaHelichrysum italicum (H. italicum) is a halophyte shrub with bright yellow flowers with a strong curry-like aroma. The essential oils of H. italicum have been used in the production of cosmetics and pharmaceuticals, due to their antiallergic and anti-inflammatory properties. In the agri-food sector, H. italicum flowers can be used for seasoning and flavoring food, and as natural food preservatives. Here, we report on the composition, bioactive compounds, and nutritive value of H. italicum flowers. Flowers were mainly composed of carbohydrates (>80 % dry weight), followed by minerals (6.31 ± 0.95 % dw), protein (5.44 ± 0.35 % dw), and lipids (3.59 % ± 0.53 % dw). High percentages of Fe, Zn, Ca, and K were found in the flower material, along with a high content in antioxidants, polyphenols, and carotenoids, as corroborated by the nuclear magnetic resonance (NMR) data. Flowers were mainly composed of saturated fatty acids (SFAs) (54.50 ± 0.95 % of total FA), followed by polyunsaturated fatty acids (PUFAs) (37.73 ± 1.25 % of total FA) and monounsaturated fatty acids (MUFAs) (7.77 ± 0.34 %), as detected by gas chromatography mass spectrometry (GC–MS). The omega-6 PUFA linoleic acid (22.55 ± 0.76 % of total FA) was the most abundant fatty acid found. Flower extracts showed antimicrobial activity against Saccharomyces cerevisiae and Komagataella phaffii, as well as against Gram-negative (Klebsiella pneumoniae) and Gram-positive (Staphylococcus aureus) bacteria. H. italicum flower material was nontoxic to human intestinal Caco-2 model cells at concentrations up to 1.0 % w/v.
- Enthalpic and entropic contributions mediate the role of disulfide bonds on the conformational stability of Interleukin-4Publication . Vaz, Daniela C.; Rodrigues, J. Rui; Sebald, Walter; Dobson, Christopher M.; Brito, Rui M. M.The role of disulfide bridges in the structure, stability, and folding pathways of proteins has been the subject of wide interest in the fields of protein design and engineering. However, the relative importance of entropic and enthalpic contributions for the stabilization of proteins provided by disulfides is not always clear. Here, we perform a detailed analysis of the role of disulfidesin the conformational stability of human Interleukin-4 (IL4), a four-helix bundle protein. In order to evaluate the contribution of two out of the three disulfides to the structure and stability of IL4, two IL4 mutants, C3T-IL4 and C24T-IL4, were used. NMR and ANS binding experiments were compatible with altered dynamics and an increase of the nonpolar solventaccessible surface area of the folded state of the mutant proteins. Chemical and thermal unfolding experiments followed by fluorescence and circular dichroism revealed that both mutant proteins have lower conformational stability than the wild-type protein. Transition temperatures of unfolding decreased 14C for C3T-IL4 and 10C for C24T-IL4, when compared toWT-IL4, and the conformational stability, at 25C, decreased 4.9 kcal/mol for C3T-IL4 and 3.2 kcal/mol for C24T-IL4. Interestingly, both the enthalpy and the entropy of unfolding, at the transition temperature, decreased in the mutant proteins. Moreover, a smaller change in heat capacity of unfolding was also observed for the mutants. Thus, disulfide bridges in IL4 play a critical role in maintaining the thermodynamic stability and core packing of the helix bundle.
- Fotopolimerização de hidrogéis de ácido hialurónicoPublication . Duarte, Stacy; Pereira, Rúben; Campos, Carlos; Vaz, Daniela C.; Paulo Flores et al.O ácido hialurónico (HA) é um polissacarídeo amplamente presente no organismo humano e constitui um material com potencial para ser incorporado em hidrogéis direcionados para aplicações biomédicas. No entanto, as fracas propriedades mecânicas, rápida degradação e libertação in vivo do HA não reticulado solúvel limita muitas aplicações clínicas diretas. Deste modo, com vista melhorar as suas caraterísticas materiais procedemos à modificação química do HA com anidrido metacrílico (MA), com formação de ácido hialurónico metacrilado (MeHA), com propriedades fotopolimerizáveis, combinado ainda na presença e ausência de agente gelificante extra, tal como o polissacarídeo natural iota-carragenina. O HA utilizado foi caracterizado através de espectroscopia de ressonância magnética nuclear (RMN) e de dicroísmo circular (CD). Os hidrogéis produzidos foram analisados via testes de absorção de água e de degradação em meio aquoso, bem como através de análise mecânica dinâmica (DMA) à compressão. O estudo conduzido permitiu verificar que, enquanto os biofilmes de HA se mostraram quebradiços e com fracas propriedades mecânicas, o MeHA não só foi capaz de formar hidrogéis, por fotopolimerização, como a adição do iota-carragenina à formulação, possibilitou a formação de IPNs (Interpenetrating Polymer Networks) e de semi-IPNs com melhores propriedades físicas e mecânicas.
- Influence of commensal bacteria on the proteolytic and antigenic profiles of INFOGEST-like digested wheat gliadinPublication . Pereira-Costa, Flávio; Domingues, Vanessa S.; Roque, Ana; Almeida, Zaida L.; Cruz, Pedro F.; Cordeiro, Rachel; Trindade, Daniela; Moura, Carla; Melo, Joana B.; Pereira, Sónia G.; Vaz, Daniela C.Introduction: Celiac disease (CeD) is a chronic autoimmune enteropathy developed by genetically predisposed individuals when exposed to gluten. Gluten gliadins, along with gut microbiota, may influence CeD onset and progression through mechanisms that remain unclear. Methods: Gliadin-degrading bacterial isolates obtained from CeD patients, and their 1st-degree relatives’ stool and blood were identified (Bacillus tropicus, Enterococcus faecalis, Micrococcus sp., Cronobacter sakazakii, Pseudomonas aeruginosa, and Serratia marcescens) and used in an INFOGEST-like protocol to simulate gliadin digestion after 4 h (digested gliadin, d-gliadin). The d-gliadin digesta were analyzed by fast protein liquid chromatography (FPLC), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), fluorescence spectroscopy, and polyclonal and monoclonal (R5 and G12) enzyme-linked immunosorbent assays (ELISA). Results and discussion: In the absence of the bacterial isolates, gliadin is poorly digested and self-assembles within 1 day into intermediate and large protein oligomers/aggregates, enriched in β-sheet structure (FTIR amide I band between 1,600 and 1,700 cm−1) and able to bind thioflavin T and Congo red. Conversely, in the presence of the bacterial isolates, gliadin is further digested, leading to an increase in protein fragments. After 4 h, the P. aeruginosa, C. sakazakii, and B. tropicus d-gliadin digesta presented a mixture of d-gliadin peptides and aggregates that showed higher antigenicity (associated with the exposure of the 5-amino acid QQPFP and 6-amino acid QPQLPY epitopes, present in the 25-mer and 33-mer, respectively) than control digestions (without bacteria), while E. faecalis led to lower antigenicity. In turn, within 24 h of incubation, all bacterial isolates led to the formation of undigested material with lower antigenicity, either due to fewer 33-mers and 25-mers in solution, or to fragment aggregation into amorphous material, not exposing antigenic sequences. Hence, intestinal flora may enhance or diminish the antigenicity of gliadin, thereby modulating the immunogenic response to gliadin/gluten.
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