Detergents modify proteinase K resistance of PrP Sc in different transmissible spongiform encephalopathies (TSEs)

Prion diseases are diagnosed by the detection of their proteinase K-resistant prion protein fragment (PrP(Sc)). Various biochemical protocols use different detergents for the tissue preparation. We found that the resistance of PrP(Sc) against proteinase K may vary strongly with the detergent used. In our study, we investigated the influence of the most commonly used detergents on eight different TSE agents derived from different species and distinct prion disease forms. For a high throughput we used a membrane adsorption assay to detect small amounts of prion aggregates, as well as Western blotting. Tissue lysates were prepared using DOC, SLS, SDS or Triton X-100 in different concentrations and these were digested with various amounts of proteinase K. Detergents are able to enhance or diminish the detectability of PrP(Sc) after proteinase K digestion. Depending on the kind of detergent, its concentration - but also on the host species that developed the TSE and the disease form or prion type - the detectability of PrP(Sc) can be very different. The results obtained here may be helpful during the development or improvement of a PrP(Sc) detection method and they point towards a detergent effect that can be additionally used for decontamination purposes. A plausible explanation for the detergent effects described in this article could be an interaction with the lipids associated with PrP(Sc) that may stabilize the aggregates.     

Sulphur amino acid requirement of laying hens and the effects of excess dietary methionine on laying performance

1. Individually‐caged laying hens were fed on maize‐soya bean meal diets containing 11.50 or 12.35 MJ ME/kg with sulphur amino acid (SAA) contents varying from 5.0 to 7.0 g/kg in period 1 (0 to 20 weeks); from 5.0 to 8.0 g/kg in period 2 (21 to 36 weeks) and from 5.0 to 10.0 g/kg in period 3 (37 to 52 weeks).

2. A SAA requirement of about 750 mg/hen d, of which about 425 mg was methionine, was found to be adequate for birds producing on average 51 g egg mass per hen d over 52 weeks. The SAA requirement was found to be higher for maximum efficiency of food utilisation than for maximum egg yield.

3. A diet containing 140 g protein/kg adequately supplemented with methionine and lysine, could sustain laying performance almost identical to that achieved on a diet containing 167 g protein/kg.

4. In diets with 140 g protein/kg the addition of 0.5 to 3.5 g dl‐methionine/kg diet in excess of the requirement did not affect egg production adversely, but food conversion efficiency was decreased.     

Molecular water motions of skim milk powder solutions during acidification studied by 17O and 1H nuclear magnetic resonance and rheology

The molecular motion of water was studied in glucono-δ-lactone-acidified skim milk powder (SMP) solutions with various pH values and dry matter contents. NMR relaxometry measurements revealed that lowering the pH in SMP solutions affected 17O and 1H T2 relaxation rates almost identically. Consequently, the present study indicates that the proteins present in the samples do not affect the 1H relaxation behavior markedly, even at relatively high SMP concentrations (15-25%). Comparison of rheological measurements and NMR measurements suggested that the collapse of κ-casein during acidification could contribute to the initial decrease in 17O and 1H relaxation rate in the pH range between 6.6 and 5.5 for 15% SMP and in the pH range between 6.6 and 5.9 for 25% SMP. However, below pH 5.5 the viscosity and 17O and 1H NMR relaxation rates did not correlate, revealing that the aggregation of casein micelles, which increases viscosity below pH 5.5, does not involve major repartitioning of water.     

Energy

The 1990s was a decade in which radical institutional restructuring was imposed from above on an energy economy which was in a stage of stable maturity following half a century of rapid development. The key plank in the reforms was the withdrawal of the state from ownership of energy production where possible (natural gas and parts of electricity), and from any non‐commercial philosophy in relation to the electricity and coal enterprises which remained state‐owned. Regulation of natural monopoly elements (pipelines, transmission and distribution lines, processing plants) was virtually eliminated under the rubric of ‘light‐handed regulation’, and the scheduling of electricity generators was moved from a planned to a market system. Underlying real trends in energy intensity, production and use were unaffected by the reforms, and the expected efficiency gains which had motivated reform were not in evidence by the end of the decade.     

Favism-inducing toxins in broad beans (Vicia faba) examination of bean extracts for pyrimidine glucosides

Methods for the isolation and identification of the pyrimidine glucoside, vicine, in extracts of broad beans, are described, By thin-layer chromatography, five fluorescing or fluorescencequenching components of the extracts are identifiable. One of them, which has been isolated from the mixture, is identified as vicine, 2:4-diamino-5:6-dihydroxypyrimidine-5-(-D-glucopyranoside).