Ricardo N. Pereira

Effects of Electric Fields on Protein Unfolding and Aggregation: Influence on Edible Films Formation

Electric fields application is receiving increased attention because of its uniform heating of liquids. The mechanisms of unfolding and aggregation of whey proteins during ohmic heating may influence properties of edible films made thereof. The aim of this work was to evaluate the effects of ohmic heating on physical and structural properties of whey protein edible films and compare them with those obtained by conventional heating. The results showed that ohmic heating determined less aggregation and lower concentration of free sulphydryls in film-forming solutions. Ohmic films were thinner, less permeable to water vapor and presented nearly the same mechanical properties of conventional films. Ohmic heating induced protein conformational changes by increasing the contents of β-sheet structures in the film network. This work emphasized the effects of ohmic heating in unfolding and aggregation mechanisms of whey proteins during heat denaturation, which determined the production of protein edible films with distinctive properties.

Death kinetics of Escherichia coli in goat milk and Bacillus licheniformis in cloudberry jam treated by ohmic heating

The influence of ohmic heating on the death kinetic parameters of Escherichia coli ATCC® 25922 in goat milk and spores of Bacillus licheniformis ATCC® 14580 in cloudberry jam was investigated and compared with that of conventional heating. Ohmic treatment of goat milk shortened the decimal reduction time D in comparison with the D values obtained at conventional treatment. Similarly, the z value, increase of temperature required for a ten-fold reduction of D, was also lower at ohmic treatment. The death kinetics of Bacillus licheniformis ATCC® 14580 spores in cloudberry jam was also studied employing both types of heat treatment. Similar conclusions were obtained for the D values as in the case of goat milk. However, the differences between the z values obtained for ohmic and conventional heating were not significant.

Thermal modification of activated carbon surface chemistry improves its capacity as redox mediator for azo dye reduction.

The surface chemistry of a commercial AC (AC(0)) was selectively modified, without changing significantly its textural properties, by chemical oxidation with HNO(3) (AC(HNO3)) and O(2) (AC(O2)), and thermal treatments under H(2) (AC(H2)) or N(2) (AC(N2)) flow. The effect of modified AC on anaerobic chemical dye reduction was assayed with sulphide at different pH values 5, 7 and 9. Four dyes were tested: Acid Orange 7, Reactive Red 2, Mordant Yellow 10 and Direct Blue 71. Batch experiments with low amounts of AC (0.1 g L(-1)) demonstrated an increase of the first-order reduction rate constants, up to 9-fold, as compared with assays without AC. Optimum rates were obtained at pH 5 except for MY10, higher at pH 7. In general, rates increased with increasing the pH of point zero charge (pH(pzc)), following the trend AC(HNO3) < AC(O2) < AC(0) < AC(N2) < AC(H2). The highest reduction rate was obtained for MY10 with AC(H2) at pH 7, which corresponded to the double, as compared with non-modified AC. In a biological system using granular biomass, AC(H2) also duplicated and increase 4.5-fold the decolourisation rates of MY10 and RR2, respectively. In this last experiment, reaction rate was independent of AC concentration in the tested range 0.1-0.6 g L(-1).

Removal of Ni(II) from aqueous solutions by an Arthrobacter viscosus biofilm supported on zeolite: From laboratory to pilot scale

This study discusses the retention of Ni(II) by Arthrobacter viscosus supported on zeolite 13 X in batch mode and in continuous mode, at laboratory scale and at pilot scale. The maximum adsorption capacities of 28.37, 20.21 and 11.13 mg/g were recorded for lab scale batch, for continuous lab scale minicolumns and for pilot scale bioreactors, respectively. The Sips isotherm and pseudo second order kinetics described well the observations registered in batch assays. The Adams-Bohart, Thomas and Yoon-Nelson models were applied to data obtained with the pilot scale bioreactor and a good fit was reached for Adams-Bohart and for Yoon-Nelson models. A fed-batch was performed at lab scale and the applicability of the biofilm in continuous mode for the described purpose was confirmed. The sorption mechanism was investigated in detail through FTIR, SEM and EDX analyses.

Exploring the Denaturation of Whey Proteins upon Application of Moderate Electric Fields: A Kinetic and Thermodynamic Study

Thermal processing often results in disruption of the native conformation of whey proteins, thus affecting functional properties. The aim of this work was to evaluate the effects of moderate electric fields on denaturation kinetics and thermodynamic properties of whey protein dispersions at temperatures ranging from 75 to 90 °C. Application of electric fields led to a lower denaturation of whey proteins, kinetically traduced by lower values of reaction order (n) and rate constant (k) (p < 0.05), when compared to those from conventional heating under equivalent heating rates and holding times. Furthermore, the application of electric fields combined with short come-up times has reduced considerably the denaturation of proteins during early stages of heating (>30% of native soluble protein than conventional heating) and has determined also considerable changes in calculated thermodynamic properties (such as E(a), ΔH(‡), ΔS(‡)). In general, denaturation reactions during moderate electric fields processing were less dependent on temperature increase.

Goat milk free fatty acid characterization during conventional and ohmic heating pasteurization.

The disruption of the milk fat globule membrane can lead to an excessive accumulation of free fatty acids in milk, which is frequently associated with the appearance of rancid flavors. Solid-phase microextraction and gas chromatography techniques have been shown to be useful tools in the quantification of individual free fatty acids in dairy products providing enough sensitivity to detect levels of rancidity in milk. Therefore, the aim of this study was to characterize the short-chain and medium-chain free fatty acid profile in i) raw untreated goat milk; ii) raw goat milk passing through pumps and heating units (plate-and-frame heat exchanger and ohmic heater); and iii) processed goat milk by conventional and ohmic pasteurization to determine the influence of each treatment in the final quality of the milk. Multivariate statistical analysis has shown that the treatments studied were not responsible for the variability found on free fatty acid contents. In particular, it was possible to conclude that ohmic pasteurization at 72 degrees C for 15 s did not promote an extended modification of free fatty acid contents in goat milk when compared with that of conventional pasteurization. Furthermore, principal component analysis showed that the capric acid can be used to discriminate goats milk with different free fatty acid concentrations. Hierarchical cluster analysis showed evidence of the existence of correlations between contents of short and medium chain free fatty acids in goat milk.

Do hypoxia/normoxia culturing conditions change the neuroregulatory profile of Wharton Jelly mesenchymal stem cell secretome?

IntroductionThe use of human umbilical cord Wharton Jelly-derived mesenchymal stem cells (hWJ-MSCs) has been considered a new potential source for future safe applications in regenerative medicine. Indeed, the application of hWJ-MSCs into different animal models of disease, including those from the central nervous system, has shown remarkable therapeutic benefits mostly associated with their secretome. Conventionally, hWJ-MSCs are cultured and characterized under normoxic conditions (21 % oxygen tension), although the oxygen levels within tissues are typically much lower (hypoxic) than these standard culture conditions. Therefore, oxygen tension represents an important environmental factor that may affect the performance of mesenchymal stem cells in vivo. However, the impact of hypoxic conditions on distinct mesenchymal stem cell characteristics, such as the secretome, still remains unclear.MethodsIn the present study, we have examined the effects of normoxic (21 % O2) and hypoxic (5 % O2) conditions on the hWJ-MSC secretome. Subsequently, we address the impact of the distinct secretome in the neuronal cell survival and differentiation of human neural progenitor cells.ResultsThe present data indicate that the hWJ-MSC secretome collected from normoxic and hypoxic conditions displayed similar effects in supporting neuronal differentiation of human neural progenitor cells in vitro. However, proteomic analysis revealed that the use of hypoxic preconditioning led to the upregulation of several proteins within the hWJ-MSC secretome.ConclusionsOur results suggest that the optimization of parameters such as hypoxia may lead to the development of strategies that enhance the therapeutic effects of the secretome for future regenerative medicine studies and applications.

Design of whey protein nanostructures for incorporation and release of nutraceutical compounds in food

ABSTRACT Whey proteins are widely used as nutritional and functional ingredients in formulated foods because they are relatively inexpensive, generally recognized as safe (GRAS) ingredient, and possess important biological, physical, and chemical functionalities. Denaturation and aggregation behavior of these proteins is of particular relevance toward manufacture of novel nanostructures with a number of potential uses. When these processes are properly engineered and controlled, whey proteins may be formed into nanohydrogels, nanofibrils, or nanotubes and be used as carrier of bioactive compounds. This review intends to discuss the latest understandings of nanoscale phenomena of whey protein denaturation and aggregation that may contribute for the design of protein nanostructures. Whey protein aggregation and gelation pathways under different processing and environmental conditions such as microwave heating, high voltage, and moderate electrical fields, high pressure, temperature, pH, and ionic strength were critically assessed. Moreover, several potential applications of nanohydrogels, nanofibrils, and nanotubes for controlled release of nutraceutical compounds (e.g. probiotics, vitamins, antioxidants, and peptides) were also included. Controlling the size of protein networks at nanoscale through application of different processing and environmental conditions can open perspectives for development of nanostructures with new or improved functionalities for incorporation and release of nutraceuticals in food matrices.

Effect of moderate electric fields in the properties of starch and chitosan films reinforced with microcrystalline cellulose

Microcrystalline cellulose (MCC) can provide improved properties when the aim is the development of biodegradable packaging materials. In this work the physicochemical properties of polysaccharide-based films (chitosan and starch) with the incorporation of MCC and the application of moderate electric field (MEF) and ultrasonic bath (UB) as treatments, were evaluated. For each treatment, the thickness, moisture content, solubility, water vapor permeability, contact angle, mechanical properties, along with its color and opacity were determined. The surface morphologies of the films were assessed by scanning electron microscopy (SEM). X-ray diffraction (XRD), Fourier-transform infrared (FTIR) spectroscopy and thermogravimetric analysis (TGA) were also performed. It was observed that the addition of different concentrations of MCC as well as the application of MEF are responsible for changes in the properties of the films, being this effect dependent on the polysaccharide used. Chitosan-based films were slightly yellow, transparent and presented a more homogeneous structure. The use of MEF was efficient in decreasing the permeability to water vapor in chitosan based films without MCC, as well as in production of films with a more hydrophobic surface. The addition of MCC promoted more opaque, rigid, less flexible and less hydrophobic films. Starch-based films were whitish, with a more heterogeneous structure and the application of MEF generated more hydrophilic films with lower tensile strength and Youngs modulus. The films with MCC were more opaque, less flexible and less hydrophilic than the films without MCC. The composites presented good thermal properties, which increases their applicability as packaging materials. Therefore, the incorporation of MCC into polysaccharide-based films as well as the application of MEF can be an approach to change the properties of films.

Comparison of chemical properties of food products processed by conventional and ohmic heating

The effect of ohmic and conventional heat processing of different food products on their chemical and physical parameters was studied. Depending on the food being analysed, parameters such as pH, total solids, ash, titratable acidity, ascorbic acid, total sugars, total fatty acids, total phenolic compounds, and anthocyanins content were determined before and after ohmic and conventional pasteurization techniques and the results were compared using one-way analysis of variance. In goat milk samples treated by ohmic technology the pH value (6.58) and total fatty acids content in milk fat (86.5 mass %) were comparable to those found in milk treated by conventional process, however, ohmically treated samples presented a lower content of lactic acid, 0.13 %. In cloudberry jam samples treated by ohmic technology the results of some of the main parameters tested, such as total sugar content 46.1 mass %, ascorbic acid content 2.83 mass %, and titratable acidity 6.01 mass % (as citric acid) did not show significant differences when compared with samples treated by conventional technology.