Abstract: Allergic reactions to food are on the rise worldwide and there is a corresponding increase in interest to understand the molecular mechanisms responsible. Peanut allergies are the most problematic because the reaction often persists into adulthood and can be as severe as anaphylaxis and death. The purpose of the work presented here was to develop a reproducible method to produce large quantities of pure recombinant Ara h 1(rAra h 1) that will enable standardization of immunological tests for patients and allow structural and immunological studies on the wild type and mutagenized forms of the protein. Ara h 1 is initially a pre-pro-protein which, following two endoproteolytic cleavages, becomes the mature form found in peanut. The mature form however has flexible regions that make it refractory to some structural studies including crystallography. Therefore, independent purification of the mature and core regions was desirable. Expression constructs were synthesized cDNA clones for each in a pET plasmid vector without tags. Codons were optimized for expression in E. coli. High-level expression was achieved in BL21 strains. Purification to near homogeneity was achieved by a combination of ammonium sulfate precipitation and ion exchange chromatography. The purified rAra h 1 was then compared with natural Ara h 1 for IgE binding. All patients recognized both the folded natural and rAra h 1, but the IgE binding to the rArah1 was significantly reduced in comparison to the natural allergen, which could potentially make it useful for immunotherapeutic purposes.
Abstract: Aim: To determine the contribution of food and vitamin D supplements on the serum 25-hydroxyvitamin D3 (25(OH)D3) concentration between October and April in a northern country (almost absent vitamin D synthesis by sunlight). Methods: Children aged 1–18 years were selected who visited the general pediatrician with a complaint whereby serum 25(OH)D3 concentration was determined. The intake of vitamin D was calculated based on a dietary questionnaire. Results: 51.1% of the 174 children had a serum 25(OH)D3 concentration below 50 nmol/L, 9.2% had a serum 25(OH)D3 concentration below 30 nmol/L. Adolescents showed lower concentrations compared to younger children. There was a positive correlation between the total amount of vitamin D obtained from food and the serum 25(OH)D3 concentration (r = 0.218, p = 0.004). The intake of milk contributed more to the serum 25(OH)D3 concentration compared to the intake of artificial supplementation, butter or fish. Conclusions: In the absence of vitamin D synthesis by sunlight, vitamin D obtained from food has a significant influence on the serum 25(OH)D3 concentration in children. Vitamin D supplements can be described as trivial. This means we should pay more attention to food as a natural source of vitamin D.
Abstract: Seafood is an important food source for many. Consumers should be entitled to an informed choice, and there is growing concern about correct composition labeling of seafood. Due to its high price, it has been shown to be vulnerable to adulteration. In the present study, we focus on moisture levels in seafood. Moisture and crude protein contents of chilled and frozen cod, pangasius, salmon, shrimp and tilapia purchased from various retail outlets in the Netherlands were examined by reference methods and the values of which were compared with the reported data from other studies in literature. Significant differences in proximate composition were determined for different species and between chilled and frozen products of the same species. Pangasius products showed the highest moisture contents in general (86.3 g/100 g), and shrimp products revealed the largest differences between chilled and frozen products. Comparison with literature values and good manufacturing practice (GMP) standards exposed that, generally, chilled pangasius, frozen pangasius and frozen shrimp products presented considerably higher moisture and lower crude protein/nitrogen contents than those found in other studies. From the GMP standards, extraneous water was estimated on average at 26 g/100 g chilled pangasius product, and 25 and 34 g/100 g product for frozen shrimp and pangasius products, respectively.
Abstract: Starch is the major component of cereal grains and starchy foods, and changes in its biophysical and biochemical properties (e.g., amylose, amylopectin, pasting, gelatinization, viscosity) will have a direct effect on its end use properties (e.g., bread, malt, polymers). The use of rapid and non-destructive methods to study and monitor starch properties, such as gelatinization, retrogradation, water absorption in cereals and starchy foods, is of great interest in order to improve and assess their quality. In recent years, near infrared reflectance (NIR) and mid infrared (MIR) spectroscopy have been explored to predict several quality parameters, such as those generated by instrumental methods commonly used in routine analysis like the rapid visco analyser (RVA) or viscometers. In this review, applications of both NIR and MIR spectroscopy to measure and monitor starch biochemical (amylose, amylopectin, starch) and biophysical properties (e.g., pasting properties) will be presented and discussed.
Abstract: Coffee is a major source of dietary antioxidants; some are present in the green bean, whereas others are generated during roasting. However, there is no single accepted analytical method for their routine determination. This paper describes the adaption of three complementary assays (Folin-Ciocalteu (FC), ABTS and ORAC) for the routine assessment of antioxidant capacity of beverages, their validation, and use for determining the antioxidant capacities of extracts from coffee beans at different stages in the roasting process. All assays showed a progressive increase in antioxidant capacity during roasting to a light roast state, consistent with the production of melanoidins having a higher antioxidant effect than the degradation of CGAs. However, the three assays gave different numbers for the total antioxidant capacity of green beans relative to gallic acid (GA), although the range of values was much smaller when chlorogenic acid (CGA) was used as reference. Therefore, although all three assays indicated that there was an increase in antioxidant activity during coffee roasting, and the large differences in responses to GA and CGA illustrate their different sensitivities to different types of antioxidant molecule.
Abstract: Beyond the main bulk components of cereals such as the polysaccharides and proteins, lower concentration secondary metabolites largely contribute to the nutritional value. This paper outlines a comprehensive protocol for GC-MS metabolomic profiling of phenolics and organic acids in grains, the performance of which is demonstrated through a comparison of the metabolite profiles of the main northern European cereal crops: wheat, barley, oat and rye. Phenolics and organic acids were extracted using acidic hydrolysis, trimethylsilylated using a new method based on trimethylsilyl cyanide and analyzed by GC-MS. In order to extract pure metabolite peaks, the raw chromatographic data were processed by a multi-way decomposition method, Parallel Factor Analysis 2. This approach lead to the semi-quantitative detection of a total of 247 analytes, out of which 89 were identified based on RI and EI-MS library match. The cereal metabolome included 32 phenolics, 30 organic acids, 10 fatty acids, 11 carbohydrates and 6 sterols. The metabolome of the four cereals were compared in detail, including low concentration phenolics and organic acids. Rye and oat displayed higher total concentration of phenolic acids, but ferulic, caffeic and sinapinic acids and their esters were found to be the main phenolics in all four cereals. Compared to the previously reported methods, the outlined protocol provided an efficient and high throughput analysis of the cereal metabolome and the acidic hydrolysis improved the detection of conjugated phenolics.