Development of a real-time PCR method for the simultaneous detection of soya and lupin mitochondrial DNA as markers for the presence of allergens in processed food

Lupin and soya are members of the Leguminosae family which are recognised as some of the richest source of vegetable proteins. Lupin- and soya-containing products are available on the EU market and could cause severe adverse reactions in allergic individuals, even if consumed at low concentrations. In this context the development of methods for reliable detection of these allergens in food products is a useful tool for the surveillance of established legislation on food labelling within the EU. This work described the development of a duplex real-time PCR method allowing the simultaneous detection of traces of lupin and soya in processed food based on a specific TaqMan® probe designed on a mitochondrial tRNA-MET gene. A set of primers and probes was designed for the amplification of a 168 and 175 bp fragment of lupin and soya mitochondrial DNA, respectively. The performance of the method was established using lupin and soya flours and cookies baked from lupin- and soya-containing dough (different concentrations and baking times). The PCR platform yielded consistent and repeatable results. The specificity of the system was tested with DNA from 28 plant species. The sensitivity of the method was suitable to detect allergenic ingredients in the low mg per kg range. Both lupin and soya at a level of 2.5 mg per kg food matrix could be detected in cookies baked at 180 °C for 10 min. The method was successfully applied to bakery (e.g. bread) and vegetarian (e.g. non-meat sausages) food products that contain or may contain soya and/or lupin as ingredient or contaminant (according to the declaration on the product label).     

Influence of Poultry Species, Muscle Groups, and NaCl Level on Strength, Deformability, and Water Retention in Heat-Set Muscle Gels

The effects of poultry species (chicken and turkey), muscle groups (breast, thigh, and drum) and NaCl concentration (1% and 2%) on a low fat gel system were compared using measurements of shear stress (strength), shear strain (deformability), and two water retention measurements. Results showed doubling NaCl tended to double stress for all species and muscle groups and increased strain by 30–50%. Cross species differences showed higher stress in turkey gels than in chicken gels with 2% NaCl. For both species, stress, strain and water retention were lowest in breast gels with both NaCl concentrations, and strain was greater in thigh gels than in drum gels with 2% NaCl.     

Steam Surface Pasteurization of Beef Frankfurters

Meat and poultry products are sources of foodborne bacterial contamination. We used flash steam heating followed by evaporative cooling to quickly reduce bacterial contamination on the surface of beef frankfurters without degrading appearance. The rate of heat transfer was studied by measuring surface temperature in a pressure chamber during steam heating. Quality changes were determined by measuring color and weight as functions of steam temperature and treatment time. Efficacy of the process was verified by microbiological experiments using L. innocua. Treatment times of 30–40 set at 115–136°C gave a 4 log reduction in bacterial counts on the surface without severely affecting color or weight. After long-term storage at 6°C and at 19°C, levels of bacteria on inoculated frankfurters remained reduced and there was no difference between treated and untreated products in color or weight.     

DNA-encoding to improve performance and allow parallel evaluation of the binding characteristics of multiple antibodies in a surface-bound immunoassay format

High affinity capture agents against protein targets are essential components for immunoassays, regardless of specific analysis format. Here, we describe the use of DNA-encoded antibodies for rapidly screening the kinetic and equilibrium binding properties of twelve commercial antibodies in a parallel analysis format using a multiplexed array of microring optical resonators. We show that DNA-encoding offers advantages in terms of antigen binding capacity, compared to covalently tethered antibodies; we also demonstrate that this linkage modality facilitates the rapid self-assembly of multiplexed arrays on account of complementarity between the DNA sequences on the antibodies and sensor array, respectively. Furthermore, DNA-encoded antibodies also allow for sensor array regeneration and reprogramming, as chaotropic agents can be used to disrupt the DNA-DNA duplexes that link the capture agents to the sensor without harming the underlying DNA on the surface, which can subsequently be reloaded with antibodies either targeting the same or different antigens.     

Thermal Decomposition of Europium Hydroxide

The thermal decomposition of europium hydroxide in an air atmosphere was investigated by means of weight-loss measurements, infrared spectroscopy, X-ray diffraction analysis, and electron microscopy. These studies showed that EU(OH)° decomposed at temperatures between 225° and 300°C into EuOOH, which was stable up to about 425°C. Between 435° and 465°C this compound decomposed into cubic Eu₂O₃, which was stable until its inversion to the high-temperature monoclinic form. X-ray diffraction data were collected for Eu(OH)₃ and EuOOH and showed that the trihydroxide has a hexagonal crystal structure and the oxyhydroxide is possibly orthorhombic. The Eu(OH)₂, EuOOH, and cubic EunOa powders contained particles up to several microns in size consisting of agglomerates of crystallites in the size range 200 to 400 A. The single monoclinic Eu₂O₃ sample studied contained crystallites whose average size was greater than 2000 A.     

Effect of Grinding on Flaw Geometry and Fracture of Glass

Fracture mechanics is combined with fracture surface analysis to analyze brittle failure of glass bars which were tested relative to the direction of grinding. Grinding essentially produces two sets of flaws from which failure occurs. In the most severe set, formed basically parallel to the grinding direction, the ratio of the average depth ( a ) to the half-width ( b ) is 0.5. In the less severe set, formed perpendicular to the grinding direction, the average a / b ratio is 1.6. In both sets the most severe flaws are generally associated with a particularly deep grinding groove or gouge. The strength reduction resulting from testing perpendicular to the grinding direction results from the larger flaw size and slightly higher stress-intensity factor resulting from the greater ellipticity of the flaws formed parallel to the grinding grooves and perpendicular to the tensile axis. Detailed analysis of these 2 sets of flaws causing failure of appropriately oriented specimens shows that (1) the fracture mirror radius, r , occurs at a constant stress-intensity level independent of flaw geometry; (2) unsymmetric fracture mirrors result from unsymmetric, irregular flaws leading to unsymmetric stress-intensity distributions; (3) is constant for semielliptical flaws; and (4) fracture energy calculated from an expression including mirror constants, the flaw-to-mirror size ratio, and the flaw geometry agrees with measured values over a wide range of a / b values.     

Swelling Control in Uranium Carbide

Fine particles were dispersed in a fuel structure to trap fission gases as very small bubbles and thereby reduce fuel swelling. The dispersions were made by adding 1.5 to 3.0 wt% W to the UC; the specimens were irradiated to a maximum of ∼2 at.% burnup. Density changes were used as the measure of swelling; there was much scatter in the results. Tungsten reduced the swelling when it was uniformly dispersed. For some samples having excess U and W or a segregated structure, W increased the swelling. The results tentatively confirm that fine particles are of value in reducing fission-gas swelling of carbide fuels.     

Strain Effects and Spalling in Alpha-Bombarded ThO2

Samples of bulk polycrystalline ThO₂ were bombarded with 5-MeV α-particles to doses between 9.4×10¹⁶ and 6.0×10¹⁷ ions/cm². The sample which received the highest dose spalled during bombardment; those receiving lower doses either did not spall or did so only after postirradiation annealing. The spalling was investigated by X-ray analysis and replica and transmission electron microscopy. It is concluded that spalling resulted from severe lattice strains at the interface between damaged and undamaged material and that sintering pores played a part in the fracture process. The role of lattice defects in initiating fracture is discussed.     

Mechanical Relaxations in Mixed Alkali Silicate Glasses: II,Discussion*

The internal friction of mixed Li-Na, Li-K, Li-Rb, Li-Cs, Na-K, Na-Rb, Na-Cs, and K-Rb silicate glasses is interpreted on the basis of the various explanations proposed for the mixed alkali effect. A direct correlation was found between the mechanical loss peak due to the stress-induced movement of the alkali ions, i.e. the alkali peak, and the electrical loss properties. The large mechanical loss peak appearing when the alkalis are mixed was attributed to an interaction between dissimilar alkali ions such that an elastic dipole is formed. The absence of a comparable electrical loss peak was interpreted as indicating that the elastic dipole is electrically inactive. In glasses containing equimolar quantities of two alkalis, the activation energy and height of the new loss peak, i.e. the mixed peak, were more closely related to the size difference than to the mass difference of the alkali ions.