Update on optimized purification and characterization of natural milk allergens

Highly purified allergens namely cow's milk alpha-lactalbumin (ALA). (Bos d 4), beta-lactoglobulin (BLG) (Bos d 5) and casein (Bos d 8) and goat's milk casein were prepared from the raw milk from a single animal with a known genetic background. Consequently the natural isoforms are limited, constant and characterized. Purification included selective precipitations and chromatographical steps. Characterization of structure and allergenic activity assessment of milk allergens were carried out using physicochemical and immunochemical methods. Taken together data demonstrated the absence of impurities and of contamination by other milk allergens in each preparation. NMR and circular dichroism analyses confirmed the native conformation and proper folding of ALA and BLG and the expected absence of folding of bovine and caprine casein. Enzyme immuno assays confirmed the native conformation of BLG and the purity and immunoreactivity of all the proteins. The allergenic activity, e. g. the IgE binding capacity, of purified proteins was identical as that of those proteins when present in milk. The purified proteins also demonstrated the ability to provoke the degranulation of humanized rat basophilic leukaemia cells. All the data thus confirm the purity, identity, structural conformation and functionality of the prepared milk allergens.     

Dynamic mechanical behaviour of random copolymers of a LC-methacrylate and octyl methacrylate

The dynamic mechanical behaviour of random copolymers of LC monomer-1-(hexyloxycarbonyl)ethyl 4-[4-(methacryloyloxy)benzoyloxy]benzoate (HB) and octyl methacrylate (OMA) was studied in the main transition and flow regions. Even though the aliphatic end groups of the side chain of HB and OMA are roughly the same, the T _g temperature of poly(HB) is ∼ 80 K higher than that of poly(OMA); this fact is due to the presence of the stiff phenyl benzoate mesogenic group in the side chain of HB. With increasing content of OMA in the copolymer the superimposed curves of the storage G′ _p and loss G′′ _p moduli at a constant temperature shift towards shorter frequencies. It has been shown that this shift is mainly due to an increase of the free volume in the copolymers with increasing content of OMA. While HB monomer shows liquid crystalline (LC) properties, its polymer (poly(HB)) and random copolymers with OMA show only isotropic thermal behaviour because no flexible spacer is present in the side chain of HB which would decouple the main chain and mesogenic group motions. This means that neither the homopolymer of HB, nor its copolymers with a flexible comonomer retain the LC properties of the starting LC monomer, HB. Received: 26 September 1996/Revised: 7 November 1996/Accepted: 7 November 1996     

Tensile stress measurements on linear and branches low-density polyethylene melts. Interpretation of results with a generalized Maxwell model

Dynamic shear experiments in the linear range of deformation and extensional tests at constant strain rate have been carried out on a linear low-density polyethylene (LLDPE) melt and on two branched low-density polyethylene (LDPE) melts with different amounts of long-chain branching. Both the dynamic shear moduli and the tensile stress obey the time–temperature superposition principle. A simple model based on a nonaffine generalized Maxwell model with two relaxation times is proposed to describe the rheological behavior in elongation of these melts. Close agreement between the model and the experimental data can be obtained by adjusting the two relaxation times and the “slip parameter” of entanglements. The variations of these parameters with strain rate and their relationship with molecular structure are discussed.     

Structural properties of CoPt films patterned using ion irradiation

Patterned CoPt films were fabricated using a combination of e-beam lithography and He⁺ ion irradiation to produce a planar array of ordered CoPt squares separated by disordered CoPt areas: (i) by molecular beam epitaxy was deposited a CoPt ordered film which corresponds to a “natural” multilayer: alternating pure cobalt and pure platinum (0 0 1) planes. (ii) The film was covered by a 300 nm thick Pt layer mask. (iii) Irradiation with appropriate ion beam and fluence disorders the CoPt film where not protected by the mask. X-ray diffraction, as well as atomic and magnetic force microscopy, is used to characterise the structural and magnetic changes in the film. The He⁺ ion irradiation does not significantly modify the surface of the CoPt film: the roughness almost remains identical (2 nm). This is promising for applications in magnetic recording technologies.     

Li-intercalation behaviour of vanadium oxide thin film prepared by thermal oxidation of vanadium metal

In order to produce thin films of crystalline V₂O₅, vanadium metal was thermally oxidised at 500 °C under oxygen pressures between 250 and 1000 mbar for 1-5 min. The oxide films were characterised by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), X-ray diffraction (XRD) and Rutherford backscattering spectrometry (RBS). The lithium intercalation performance of the oxide films was investigated by cyclic voltammetry (CV), chronopotentiometry and electrochemical impedance spectroscopy (EIS). It was shown that the composition, the crystallinity and the related lithium intercalation properties of the thin oxide films were critically dependent on the oxidation conditions. The formation of crystalline V₂O₅ films was stimulated by higher oxygen pressure and longer oxidation time. Exposure for 5 min at 750 mbar O₂ at 500 °C resulted in a surface oxide film composed of V₂O_5, and consisting of crystallites up to 200 nm in lateral size. The thickness of the layer was about 100 nm. This V₂O₅ oxide film was found to have good cycling performance in a potential window between 3.8 and 2.8 V, with a stable capacity of 117 ± 10 mAh/g at an applied current density of 3.4 μA/cm². The diffusion coefficients corresponding to the two plateaus at 3.4 and 3.2 V were determined from the impedance measurements to (5.2 and 3.0) × 10⁻¹³ cm² s⁻¹, respectively. Beneath the V₂O₅ layer, lower oxides (mainly VO₂) were found close to the metal. At lower oxygen pressure and shorter exposure times, the oxide films were less crystalline and the amount of V⁴⁺ increased in the surface oxide film, as revealed by XPS. At intermediate oxygen pressures and exposure times a mixture of crystalline V₂O₅ and V₆O₁₃ was found in the oxide film.     

Effect of added dietary fat and bovine somatotropin on the performance and metabolism of lactating dairy cows

Four early lactation Holstein cows (44 to 105 d postpartum) were used in a 4 X 4 Latin square experiment in a 2 X 2 factorial arrangement of treatments to study effects of added dietary fat and/or bovine somatotropin on performance and metabolism. Treatments were: 1) control diet plus placebo injection; 2) 5% added dietary fat (hydrolyzed blend of animal and vegetable fat) + placebo injection; 3) control diet + 50 IU bovine somatotropin/d; and 4) 5% added dietary fat + 50 IU bovine somatotropin/d. Dietary fat reduced dry matter intake (21.6 vs. 22.7 kg/d) and elevated plasma triglycerides (34.7 vs. 29.2 mg/100 ml). Injection of somatotropin lowered blood urea nitrogen, increased plasma free fatty acids, and increased plasma somatotropin. Milk production, milk fat percent, and 4% FCM production were increased by the injection of somatotropin. Milk protein percent was decreased (3.30 vs. 3.44%) with added fat and tended to be lower with somatotropin. The percentage of short-chain fatty acids (C6 to C14) in milk fat decreased with added fat while the percentage of stearic and oleic acids in milk fat increased. Production responses for fat plus somatotropin and somatotropin treatments were similar. Under the conditions of this study, the addition of dietary fat with injection of somatotropin had little effect on production parameters compared with that found with somatotropin alone.     

Atopic Dermatitis: The Fate of the Fat

Atopic dermatitis (AD) is a chronic and relapsing inflammatory skin disease in which dry and itchy skin may develop into skin lesions. AD has a strong genetic component, as children from parents with AD have a two-fold increased chance of developing the disease. Genetic risk loci and epigenetic modifications reported in AD mainly locate to genes involved in the immune response and epidermal barrier function. However, AD pathogenesis cannot be fully explained by (epi)genetic factors since environmental triggers such as stress, pollution, microbiota, climate, and allergens also play a crucial role. Alterations of the epidermal barrier in AD, observed at all stages of the disease and which precede the development of overt skin inflammation, manifest as: dry skin; epidermal ultrastructural abnormalities, notably anomalies of the lamellar body cargo system; and abnormal epidermal lipid composition, including shorter fatty acid moieties in several lipid classes, such as ceramides and free fatty acids. Thus, a compelling question is whether AD is primarily a lipid disorder evolving into a chronic inflammatory disease due to genetic susceptibility loci in immunogenic genes. In this review, we focus on lipid abnormalities observed in the epidermis and blood of AD patients and evaluate their primary role in eliciting an inflammatory response.     

Three-dimensional analysis of boreholes considering spatial variability of properties and poroelastoplasticity

In general, analysis of oil producing wells is carried out considering that both hydraulic and mechanical parameters of the rock mass are deterministic. Mechanical and hydraulic properties of rock masses and in particular sedimentary rock masses may show a considerable degree of spatial variability. This paper focuses on the evaluation of the three-dimensional borehole response, particularly of the plastic zone, taking into account the spatial variability of both hydraulic and mechanical properties. The analysis is performed with a developed finite element program that incorporates spatial variability, coupled fluid-mechanical effects and elastoplastic behavior of the rock mass. Examples are shown and conclusions are drawn regarding the effect of spatial variability on the three-dimensional borehole analysis.