46 resultados para Dextransucrase assays
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Membrane-like scaffolds are suitable to induce regeneration in many and different anatomic sites, such as periodontal membrane, skin, liver and cardiac tissues. In some circumstances, the films should adapt to geometrical changes of the attached tissues, such as in cardiac or blood vessel tissue engineering applications. In this context, we developed stretchable two-dimensional multilayer constructs through the assembling of two natural-based polyelectrolytes, chitosan (CHT) and chondroitin sulphate (CS), using the layer-by-layer methodology. The morphology, topography and the transparency of the films were evaluated. The in- fluence of genipin, a natural-derived cross-linker agent, was also investigated in the control of the mechanical properties of the CHT/CS films. The water uptake ability can be tailored by changing the cross-linker concentration, which influenced the young modulus and ultimate tensile strength. The maximum extension tends to decrease with the increase of genipin concentration, compromising the elastic properties of CHT/CS films: nevertheless using lower cross-linker contents, the ultimate tensile stress is similar to the films not cross-linked but exhibiting a significant higher modulus. The in vitro biological assays showed better L929 cell adhesion and proliferation when using the crosslinked membranes and confirmed the non-cytotoxicity of the CHT/CS films. The developed free-standing biomimetic multilayer could be designed to fulfill specific therapeutic requirements by tuning properties such as swelling, mechanical and biological performances.
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Tese de Doutoramento em Ciências da Saúde
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Doctoral Dissertation for PhD degree in Chemical and Biological Engineering
Simultaneous detection of cyclopiazonic acid and aflatoxin B1 by HPLC in methanol/water mobile phase
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A simple procedure for the simultaneous detection of cyclopiazonic acid (CPA) and aflatoxin B1 from fungal extracts is presented, using a methanol and water mobile phase and fluorescence detection. This methodology has been tested with standard solutions of both mycotoxins CPA and Aflatoxin B1 and with methanolic extracts of Aspergillus section Flavi strains, previously characterized for their mycotoxin production profile. Previously available methodology required the use of two different chromatographic runs for these mycotoxins, with distinct columns and detectors (fluorescence detection with a post-column photochemical derivatization (PHRED) for aflatoxin B1 and UV detection for CPA). The proposed method detects both mycotoxins in a single run. Data from these assays will be presented and discussed.
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The presence of mycotoxins in foodstuff is a matter of concern for food safety. Wines can also be contaminated with these toxicants. Several authors have demonstrated the presence of mycotoxins in wine, especially ochratoxin A (OTA) [1]. As these toxicants can never be completely removed from the food chain, many countries have defined levels in food in order to attend health concerns. The maximum acceptable level of OTA in wines is 2.0 µg/kg according to the Commission regulation No. 1881/2006 [2]. Although, higher levels of OTA have been detected in several wine samples. In order to reduce OTA to safer levels, several oenological products can be used in wine; including activated carbons, as shown in previous experiments. Regarding this, the aim of present study was to evaluate the effectiveness of several activated carbons for reducing the amount of OTA present in white and red wines as well as to evaluate their effect on wines physicochemical characteristics. Wine samples were artificially supplemented with OTA at a final concentration of 10.0 µg/L. The different activated carbons were applied at the concentration recommended by the manufacturer in order to evaluate their efficiency in reducing OTA levels. A mixture composed by gelatine, bentonite and activated carbon reduced 80% of OTA concentration in white wine. The same mixture was however less efficient in red wine, achieving only a reduction of 55%. Thereafter, the effect of activated carbon was evaluated in a red wine, achieving reductions of 66%. Considering these results more assays are being performed with other commercial activated carbons, in order to evaluate their efficiency. These results may provide valuable information for winemakers. Knowing the effect of commercial activated carbons they may choose most appropriate products to remove OTA, thus enhancing wine safety and quality.
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The preclinical development of nanomedicines raises several challenges and requires a comprehensive characterization. Among them is the evaluation of the biodistribution following systemic administration. In previous work, the biocompatibility and in vitro targeting ability of a glycol chitosan (GC) based nanogel have been validated. In the present study, its biodistribution in the mice is assessed, using near-infrared (NIR) fluorescence imaging as a tool to track the nanogel over time, after intravenous administration. Rapid whole body biodistribution of both Cy5.5 labeled GC nanogel and free polymer is found at early times. It remains widespreadly distributed in the body at least up to 6 h postinjection and its concentration then decreases drastically after 24 h. Nanogel blood circulation half-life lies around 2 h with the free linear GC polymer presenting lower blood clearance rate. After 24 h, the blood NIR fluorescence intensity associated with both samples decreases to insignificant values. NIR imaging of the organs shows that the nanogel had a body clearance time of 48 h, because at this time point a weak signal of NIR fluorescence is observed only in the kidneys. Hereupon it can be concluded that the engineered GC nanogel has a fairly long blood circulation time, suitable for biomedical applications, namely, drug delivery, simultaneously allowing efficient and quick body clearance.
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Dissertação de mestrado em Ecologia
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Dissertação de mestrado em Biofísica e Bionanossistemas
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Dissertação de mestrado em Biofísica e Bionanossistemas
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Dissertação de mestrado em Genética Molecular
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Dissertação de mestrado em Genética Molecular
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Dissertação de Mestrado em Química Medicinal
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Dissertação de mestrado em Ecology
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The increase in heavy metal contamination in freshwater systems causes serious environmental problems in most industrialized countries, and the effort to find ecofriendly techniques for reducing water and sediment contamination is fundamental for environmental protection. Permeable barriers made of natural clays can be used as low-cost and eco-friendly materials for adsorbing heavy metals from water solution and thus reducing the sediment contamination. This study discusses the application of permeable barriers made of vermiculite clay for heavy metals remediation at the interface between water and sediments and investigates the possibility to increase their efficiency by loading the vermiculite surface with a microbial biofilm of Pseudomonas putida, which is well known to be a heavy metal accumulator. Some batch assays were performed to verify the uptake capacity of two systems and their adsorption kinetics, and the results indicated that the vermiculite bio-barrier system had a higher removal capacity than the vermiculite barrier (?34.4 and 22.8 % for Cu and Zn, respectively). Moreover, the presence of P. putida biofilm strongly contributed to fasten the kinetics of metals adsorption onto vermiculite sheets. In open-system conditions, the presence of a vermiculite barrier at the interface between water and sediment could reduce the sediment contamination up to 20 and 23 % for Cu and Zn, respectively, highlighting the efficiency of these eco-friendly materials for environmental applications. Nevertheless, the contribution of microbial biofilm in open-system setup should be optimized, and some important considerations about biofilm attachment in a continuous-flow system have been discussed.
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Dissertação de mestrado em Bioengenharia