Characterization of plasma protein gels by means of image analysis

In this study, image analysis is presented as a powerful tool to analyse changes in the microstructural properties of protein gels. Several methods were used to elucidate changes in plasma protein gels induced by changes in the pH conditions, visually noticeable but difficult to assess objectively. Fractal analysis was used to measure the degree of self-similarity, lacunarity analysis to study the distribution of pores within the protein network and the spatial variability of SEM images to describe the compactness of the gel. It was found that the lacunarity measurements and the cut-off of the autocorrelation functions complemented the fractal dimension (D_f) and the average pore size, the most used parameters in describing the microstructure of protein gels. Correlations between macroscopic properties and microstructural attributes revealed that an increase in textural attributes, such as hardness, springiness and cohesiveness, and in the capacity of gels to retain water, entailed an increase of D_f and the compactness of gels, but a decrease in the pore size and the heterogeneity of gaps.     

Physical and mechanical properties of peanut protein films

The properties of peanut protein films were modified using physical and chemical treatments, and their effects on color, mechanical strength, water solubility and barrier to water vapor and oxygen of the films were investigated. Physical treatments consisted of heat denaturation of film-forming solution for 30 min at 60°C, 70°C, 80°C and 90°C, ultraviolet irradiation of films for up to 24 h, and three ultrasound processes of film-forming solution. Chemical treatments consisted of addition of aldehydes and anhydrides. Heat curing at 70°C, ultraviolet irradiation for 24 h, ultrasound for 10 min in a water-bath, and formaldehyde and glutaraldehyde addition caused a significant increase in the tensile strength of the films. The water vapor permeability (WVP) and oxygen permeability (OP) of the films decreased after heat denaturation and aldehyde treatment. OP also decreased with UV treatment. Heat curing was the most effective treatment, making the films stronger, more resistant to water and less permeable to water vapor and oxygen.     

Polymerase chain reaction coupled with peptide nucleic acid high-performance liquid chromatography for the sensitive detection of traces of potentially allergenic hazelnut in foodstuffs

A new DNA extraction method suitable for a wide variety of complex food matrices has been devised and applied in combination with a new polymerase chain reaction (PCR) method for the sensitive detection of a specific DNA sequence univocally identifying the presence of potentially allergenic hazelnut (Corylus avellana). A 156 base pair amplicon corresponding to an internal region of the complementary DNA of the major hazelnut allergen (Cor a 1) was designed, and was found to be highly specific; the method was tested on both pure and complex food matrices and was found to be able to confirm the presence of hazelnut down to 5 pg of its DNA. The sequence of the amplicon was further confirmed by high-performance liquid chromatography (HPLC) analysis with a specific peptide nucleic acid (PNA) probe. A 15-mer PNA probe was expressly designed and synthesized to hybridize an internal sequence of the previously described amplicon. Given its reported sequence specificity and high hybridization efficiency, the PNA probe was used to develop an anion-exchange HPLC method allowing for a fast and reliable confirmation of the identity of the amplified products. The PCR–HPLC method was successfully tested on commercial samples, allowing for the detection of the presence of potentially hidden allergens even in products where the presence of hazelnut as an ingredient or possible contaminant was not reported.     

Parameters for the evaluation of the thermal damage and nutraceutical potential of lupin-based ingredients and food products

Foods based on sweet lupin proteins are gaining attention from industry and consumers because of their possible role in the prevention of cardiovascular disease. When promoting lupin-based foods for inclusion in a daily diet, the thermal damage suffered during processing is of relevance to the bioactive and nutritional quality of the food product. N-(2-furoylmethyl)-L-lysine (furosine) quantification demonstrates that currently available sweet lupin protein isolates have a thermal damage comparable to or lower than other traditional food ingredients, and are a good source of lysine in non-dairy products. In lupin-based foods claiming to have cholesterol-lowering potential, shotgun proteomics offers itself as a fast and effective screening method for assessing the biological availability of active peptides. Such a method is readily applicable to other legume-enriched food products.     

Optimization of a gelled emulsion intended to supply ω-3 fatty acids into meat products by means of response surface methodology

The optimization of a gelled oil-in-water emulsion was performed for use as fat replacer in the formulation of ω-3 PUFA-enriched cooked meat products. The linseed oil content, carrageenan concentration and surfactant–oil ratio were properly combined in a surface response design for maximizing the hardness and minimizing the syneresis of the PUFA delivery system. The optimal formulation resulted in a gelled emulsion containing 40% of oil and 1.5% of carrageenan, keeping a surfactant–oil ratio of 0.003. The gel was applied as a partial fat replacer in a Bologna-type sausage and compared to the use of an O/W emulsion also enriched in ω-3. Both experimental sausages contributed with higher ω-3 PUFA content than the control. No sensory differences were found among formulations. The selected optimized gelled oil-in-water emulsion was demonstrated to be a suitable lipophilic delivery system for ω-3 PUFA compounds and applicable in food formulations as fat replacer.     

Healthy reduced-fat Bologna sausages enriched in ALA and DHA and stabilized with Melissa officinalis extract

Reduced-energy and reduced-fat Bologna products enriched with ω-3 polyunsaturated fatty acids were formulated by replacing the pork back-fat by an oil-in-water emulsion containing a mixture of linseed-algae oil stabilized with a lyophilized Melissa officinalis extract. Healthier composition and lipid profile was obtained: 85 kcal/100 g, 3.6% fat, 0.6 g ALA and 0.44 g DHA per 100 g of product and ω-6/ω-3 ratio of 0.4. Technological and sensory problems were not detected in the new formulations. Reformulation did not cause oxidation problems during 32 days of storage under refrigeration. The results suggest that it is possible to obtain reduced-fat Bologna-type sausages rich in ALA and DHA and stabilized with natural antioxidants, applying the appropriate technology without significant effects on the sensory quality, yielding interesting products from a nutritional point of view.     

Study of OH Radicals in Human Serum Blood of Healthy Individuals and Those with Pathological Schizophrenia

The human body is constantly under attack from free radicals that occur as part of normal cell metabolism, and by exposure to environmental factors such as UV light, cigarette smoke, environmental pollutants and gamma radiation. The resulting “Reactive Oxygen Species” (ROS) circulate freely in the body with access to all organs and tissues, which can have serious repercussions throughout the body. The body possesses a number of mechanisms both to control the production of ROS and to cope with free radicals in order to limit or repair damage to tissues. Overproduction of ROS or insufficient defense mechanisms leads to a dangerous disbalance in the organism. Thereby several pathomechanisms implicated in over 100 human diseases, e.g., cardiovascular disease, cancer, diabetes mellitus, physiological disease, aging, etc., can be induced. Thus, a detailed investigation on the quantity of oxygen radicals, such as hydroxyl radicals (OH^•) in human serum blood, and its possible correlation with antioxidant therapy effects, is highly topical. The subject of this study was the influence of schizophrenia on the amount of OH^• in human serum blood. The radicals were detected by fluorimetry, using terephthalic acid as a chemical trap. For all experiments the serum blood of healthy people was used as a control group.     

Mechanistic Investigation of ROS-Induced DNA Damage by Oestrogenic Compounds in Lymphocytes and Sperm Using the Comet Assay

Past research has demonstrated that oestrogenic compounds produce strand breaks in the DNA of sperm and lymphocytes via reactive oxygen species (ROS). In the current investigation, sperm and lymphocytes were treated in vitro with oestrogenic compounds (diethylstilboestrol, progesterone, 17β-oestradiol, noradrenaline and triiodotyronine) and several aspects of DNA damage were investigated. Firstly, mediation of DNA damage by lipid peroxidation was investigated in the presence of BHA (a lipid peroxidation blocker). BHA reduced the DNA damage generated by 17β-oestradiol and diethylstilboestrol in a statistically significant manner. No effects were observed for sperm. Secondly, the presence of oxidized bases employing FPG and EndoIII were detected for lymphocytes and sperm in the negative control and after 24 h recovery in lymphocytes but not immediately after treatment for both cell types. The successful detection of oxidized bases in the negative control (untreated) of sperm provides an opportunity for its application in biomonitoring studies. DNA repair at 24 h after exposure was also studied. A nearly complete recovery to negative control levels was shown in lymphocytes 24 h recovery after oestrogenic exposure and this was statistically significant in all cases. Rapid rejoining of DNA, in a matter of hours, is a characteristic of DNA damaged by ROS.     

Determination of hemin-binding characteristics of proteins by a combinatorial peptide library approach

Studies of the binding of heme/hemin to proteins or peptides have recently intensified as it became evident that heme serves not only as a prosthetic group, but also as a regulator and effector molecule interacting with transmembrane and cytoplasmic proteins. The iron‐ion‐containing heme group can associate with these proteins in different ways, with the amino acids Cys, His, and Tyr allowing individual modes of binding. Strong coordinate‐covalent binding, such as in cytochrome c, is known, and reversible attachment is also discussed. Ligands for both types of binding have been reported independently, though sometimes with different affinities for similar sequences. We applied a combinatorial approach using the library (X)₄(C/H/Y)(X)₄ to characterize peptide ligands with considerable hemin binding capacities. Some of the library‐selected peptides were comparable in terms of hemin association independently of whether or not a cysteine residue was present in the sequence. Indeed, a preference for His‐based (≈39 %) and Tyr‐based (≈40 %) sequences over Cys‐based ones (≈21 %) was detected. The binding affinities for the library‐selected peptides, as determined by UV/Vis spectroscopy, were in the nanomolar range. Moreover, selected representatives efficiently competed for hemin binding with the human BK channel hSlo1, which is known to be regulated by heme through binding to its heme‐binding domain.     

Total asymmetric synthesis of monosaccharides and analogues

Since the discovery of the 'formose reaction' by Butlerow, total synthesis of carbohydrates has undergone rapid development. The most important methods for the asymmetric synthesis of monosaccharides and analogues of biological importance are presented. Nowadays any natural and non-natural monosaccharide can be prepared pure in both enantiomeric forms starting from inexpensive starting materials. Metal-based asymmetric catalysis and organocatalysis have been successfully applied, alone or in combination with chemoenzymatic methods. Alternative methods rely upon substrate- or reagent-controlled diastereo- and enantioselective reactions. Suitably protected carbohydrates have been prepared by total synthesis, thus allowing their direct use in the preparation of oligosaccharides and analogues.