Reduction of immunoreactivity of bovine beta-lactoglobulin upon combined physical and proteolytic treatment

Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin before, during and after treatment at 600 MPa and pH 6.8 for 10 min at 30, 37 and 44 degrees C. The extent of beta-lactoglobulin hydrolysis under pressure was noticeably higher than at atmospheric pressure, particularly when chymotrypsin was used. Addition of proteases at ambient pressure to previously pressure-treated beta-lactoglobulin gave only a modest increase in proteolysis with respect to the untreated protein. Products of enzyme hydrolysis under pressure were separated by reverse-phase HPLC, and were found to be different from those obtained at atmospheric pressure when chymotrypsin was used. The residual immunochemical reactivity of the products of combined pressure-enzyme treatment was assessed on the unresolved hydrolysates by ELISA tests using polyclonal and monoclonal antibodies, and on individual hydrolytic fractions by Western Blotting using sera of paediatric patients allergic to whey proteins in cow milk. The immunoreactivity of the whole hydrolysates was related to their content of residual intact beta-lactoglobulin, and no immunochemical reactivity was found for all the products of chymotrypsin hydrolysis under pressure. The results indicate that chymotrypsin effectively hydrolysed hydrophobic regions of beta-lactoglobulin that were transiently exposed during the pressure treatments and that were not accessible in the native protein or in the protein that had been previously pressure treated.     

Immunochemical detection of ovalbumin in dairy-based foods

Commercially available antibodies were used for the detection and quantitation of ovalbumin in ice cream and cheese. Electrophoretic separation of protein components, followed by blotting and immunochemical recognition of ovalbumin allowed the detection of the addition of 50 g kg^-1 egg white to processed cheese and allowed detection of egg in ice cream. For quantitative purposes a competitive ELISA method was set up. Calibration curves for ovalbumin in various products were obtained. A marked matrix effect was evident in ice cream, cheese and yoghurt. Different types of cheese gave nearly identical matrix effects. Analytical application of the method should therefore be possible, at least within a given class of products. A study on the influence of thermal treatments on the ELISA response showed that melting of cheese at various temperatures modifies only slightly the ELISA response for ovalbumin.     

Molecular basis of open-angle glaucoma in Italy

Glaucoma is a group of ocular diseases characterized by an optic neuropathy in which degeneration of retinal ganglion cells leads to a characteristic excavation of the optic nerve head. Primary open-angle glaucoma (POAG) can be subdivided into two groups according to age of onset:- 1. the more common middle- to late-age onset, chronic open-angle glaucoma (COAG) diagnosed after the age of 40 years; 2. the rarer juvenile open-angle glaucoma (JOAG), which is diagnosed between the age of 3 years and early adulthood. Recently, the gene coding for the trabecular meshwork-induced glucocorticoid response protein (TIGR), located in chromosome 1 (1q23-25), was found mutated in patients affected by POAG. In this work we describe the clinical and molecular genetic features of several Italian families affected by autosomal dominant POAG, collected in various regions of Italy.     

Reversible and non-denaturing replacement of iron by cadmium in Clostridium pasteurianum ferredoxin

Incubation of native, reduced Clostridium pasteurianum ferredoxin with different metals gave a range of modifications in the electronic and EPR spectrum of the protein, or made the signals disappear. The reduced protein, isolated after incubation with different metals under identical conditions (50 microM protein, 1 mM metal, 1 h incubation) was found to contain amounts of foreign metals increasing with their thiophylicity, i.e. Cd2+ > Zn2+ > Co2+. Little, if any, incorporation was observed for Ni2+, Cu2+, Mn2+ or in the absence of reductant. The activity of substituted ferredoxins in a hydrogenase-coupled assay was proportional to the amount of residual iron, suggesting that the residual iron is present in a population of intact active molecules rather than in partially substituted clusters distributed among individual molecules. The cadmium-substituted ferredoxin did not contain iron, but contained eight cadmium atoms and six labile sulfide atoms/mol. Folding of the isolated, substituted proteins was investigated by CD and 1H-NMR. Both techniques showed retention of the main structural features of the protein upon metal substitution. The rate and extent of the substitution of iron by cadmium were essentially independent of pH, but were found to decrease with increasing ionic strength and to increase with the cadmium concentration. In the cadmium-substituted protein, cadmium was replaced by iron upon incubation with iron and mercaptoethanol in the absence of dithionite. In the presence of dithionite, cadmium was not replaced by iron upon incubation of the cadmium-substituted protein with excess iron and mercaptoethanol. In competition experiments, incubation of iron-containing ferredoxin with stoichiometric amounts of cadmium in the presence of dithionite and excess iron and mercaptoethanol resulted in quantifiable replacement of iron by cadmium. Therefore, substitution of iron by cadmium was only achieved under reducing conditions, and was only reversible in the absence of strong reductants.     

Théorie de l’information. Structure et modélisation d’un réseau de connexion

The authors study the switching network structures with emphasis on the information theory aspects of the problem (definition of combinatorial power and complexity). Different routing rules and rearranging algorithms are surveyed. Finally, a numerical simulation points out the extensivity concept which is useful in the study of switching networks models of reduced size.     

Conductivities of calcium chloride and calcium chloride-sodium chloride solutions saturated in calcium carbide and unsaturated in calcium oxide

The specific conductivity variation of the named solutions vs temperature in the range 750–950°C and vs the sodium chloride molar fractions have been measured. The specific conductivity increases sharply with increasing sodium chloride M fraction and reaches a maximum when the M fraction ratio of sodium chloride to calcium carbide is nearly two. This experimental result has been interpreted by assuming interactions between calcium carbide and sodium chloride, and constitutes a verification of a previous thermodynamic result obtained by the same authors.