Measurement of the oxidation-reduction potential of cheddar cheese.

The objective of this study was to develop a method to measure the oxidation-reduction (redox) potential of hard cheeses such as cheddar and to investigate the impact on this parameter of measurement temperature, and factors associated with electrochemical cell design such as distance between reference and working electrodes and depth into the cheese of the platinum electrodes. For this purpose, a novel, self-sealing, platinum working electrode was constructed which was thin and flexible enough to be inserted directly into the cheese sample. A calomel electrode was used as the reference electrode and the circuit was completed with a 3 M KCl salt bridge. The physical orientation of electrodes, such as distance between reference electrode and working electrode, had a substantial effect on equilibrium time for redox potential measurement. The time required for redox potential to reach equilibrium was 2 d in cheddar cheese and the optimum distance between the platinum and calomel electrodes was 2.5 cm. The fastest equilibration time was obtained when the working electrode was inserted 5 or 6 cm into the cheese. Temperature also had an important effect on redox potential. The shortest time to reach equilibrium of potential was at room temperature (20 degrees C), but it was not practical to keep cheese at this temperature for a period of 2 d. Therefore, redox measurement at 12 degrees C was recommended in spite of the longer equilibration time compared with room temperature. The results of this study suggest that the novel platinum working electrode allows reproducible measurement of the oxidation-reduction potential of cheddar cheese.

Effects of mineral salts and calcium chelating agents on the gelation of renneted skim milk.

The effects of adding CaCl2, orthophosphate, citrate, EDTA, or a mixture of these, to reconstituted skim milk (90 g of solids/kg solution) on the gelation of renneted milk were mediated by changes in Ca2+ activity and the casein micelle. At pH 6.65, the addition of citrate or EDTA, which removed more than 33% of the original colloidal calcium phosphate with the accompanying release of 20% casein from the micelle, completely inhibited gelation. Reformation of the depleted colloidal calcium phosphate and casein in the micelle, by the addition of CaCl2, removed this inhibition. When the minimum requirements for colloidal calcium phosphate and casein in the micelle were met, the coagulation time decreased with increasing Ca2+ activity, leveling off at high Ca2+ activity. The storage modulus of renneted gels, measured at 3 h, increased with increasing colloidal calcium phosphate content of micelles up to a level at which it was approximately 130% of the original colloidal calcium phosphate in the micelles. Further increases in colloidal calcium phosphate by the addition of CaCl2, orthophosphate, or mixtures of these, which did not change the proportion of casein in the micelle, decreased the storage modulus. The gelation of the renneted milk was influenced by Ca2+ activity, the amounts of colloidal calcium phosphate, and casein within the micelle, with the effects of colloidal calcium phosphate and casein within the micelle clearly dominating the storage modulus. These results are consistent with the model of Horne (Int. Dairy J. 8:171-177, 1998) which postulates that, following cleavage of the stabilizing K-casein hairs by rennet, the properties of the rennet gel are determined by the balance between the electrostatic and hydrophobic forces between casein micelles.

Mineral and casein equilibria in milk: effects of added salts and calcium-chelating agents.

We have investigated the effects of adding a range of mineral salts and calcium-chelating agents on the distribution of casein and minerals between the non-pelleted and pelleted phases of milk obtained upon centrifugation at 78000 g for 90 min. Adding CaCl2 or mixtures of NaH2PO4 and Na2HPO4 to reconstituted skim milk (90 g milk solids/kg) at pH 6.65 increased both pelleted casein and pelleted calcium phosphate. Opposite effects were obtained by adding citrate or EDTA. The change in pelleted calcium phosphate was not simply related to casein release from the micelle. Upon adding 5 mmol EDTA/kg milk, 20% of the pelleted Ca, 22% of the pelleted phosphate and 5% of the micellar casein were removed. Increasing the concentration of EDTA to 10 mmol/kg milk decreased the pelleted Ca by 44% and the pelleted phosphate by 46%, and caused 30% of the micellar casein to be released. The effects of adding phosphate, citrate or EDTA at pH 6.65, followed by the addition of CaCl2, demonstrated the reversibility of the dissolution and formation of the micellar calcium phosphate. There were limits to this reversibility that were related to the amount of colloidal calcium phosphate removed from the casein micelles. Adding CaCl2 to milk containing > or = 20 mmol EDTA or > or = 30 mmol citrate/kg milk did not result in complete reformation of casein micelles. Light-scattering experiments confirmed that the dissolution of moderate amounts of colloidal calcium phosphate had little effect on micellar size and were reversible, while the dissolution of larger amounts of colloidal calcium phosphate resulted in large reductions in micellar size and was irreversible.

A twin titration microcalorimeter for the study of biochemical reactions.

A small-volume (200 microliter) titration calorimeter of high sensitivity (1 mu cal ) has been developed for the purpose of studying biochemical reactions where the amounts of material are limited to a few nanomoles. High sensitivity is achieved by calorimetric twining , use of glass cells, elimination of vapor space, effective low-energy stirring, and reduction of measurement time. The calorimeter operates using the heat conduction principal with computer-controlled electrical compensation, which reduces the measurement time of each point from 10 to 3 min. This reduction in time is accompanied by a corresponding increase in the precision of measurement. The use of the calorimeter is demonstrated by a measurement of the heat of oxygenation of hemocyanin.

The effect of 1,4-diaminobutanone on the stability of ornithine decarboxylase from Aspergillus nidulans.

1,4-Diaminobutanone, a competitive inhibitor of ornithine decarboxylase in Aspergillus nidulans, is able to increase the half-life of this enzyme and thus stimulate an increase in its activity in vivo. It also protects ornithine decarboxylase against proteolysis by chymotrypsin in vitro.

An anaphylactoid reaction to gallamine triethiodide.

A patient suffered profound anaphylactoid reaction during anaesthesia. Intradermal skin testing confirmed the clinical impression that gallamine caused the reaction. The method of testing is discussed. Previous reports of hypersensitivity to the drug are reviewed, and the use of adrenaline, antihistamines and corticosteroids in patient management are considered.

Polyamine and ornithine metabolism during the germination of conidia of Aspergillus nidulans.

1. The activities of ornithine decarboxylase, S-adenosylmethionine decarboxylase and ornithine-2-oxoglutarate aminotransferase were studied during the first 24 h of conidial germination in Aspergillus nidulans. 2. Increases (over 100-fold) in the activities of ornithine decarboxylase and S-adenosylmethionine decarboxylase occurred during the emergence of the germ-tube and before the doubling of DNA and this was followed by a sharp fall in the activities of both enzymes by 16h. 3. The increase in ornithine decarboxylase could be largely suppressed if 0.6 mM-putrescine was added to the growth medium. 4. Low concentrations of cycloheximide, which delayed germination by 2h, caused a corresponding delay in the changes in ornithine decarboxylase activity. 5. Ornithine-2-oxoglutarate aminotransferase activity increased steadily during the first 24h of germination. 6. Ornithine or arginine in the growth medium induced higher activity of ornithine-2-oxoglutarate aminotransferase, but did not affect ornithine decarboxylase activity. 7. The significance of these enzyme changes during germination is discussed.