Analysis and mapping of CACNB4, CHRNA1, KCNJ3, SCN2A and SPG4, physiological candidate genes for porcine congenital progressive ataxia and spastic paresis

The cause of porcine congenital progressive ataxia and spastic paresis (CPA) is unknown. This severe neuropathy manifests shortly after birth and is lethal. The disease is inherited as a single autosomal recessive allele, designated cpa. In a previous study, we demonstrated close linkage of cpa to microsatellite SW902 on porcine chromosome 3 (SSC3), which corresponds syntenically to human chromosome 2. This latter chromosome contains ion channel genes (Ca(2+), K(+) and Na(+)), a cholinergic receptor gene and the spastin (SPG4) gene, which cause human epilepsy and ataxia when mutated. We mapped porcine CACNB4, KCNJ3, SCN2A and CHRNA1 to SSC15 and SPG4 to SSC3 with the INRA-Minnesota porcine radiation hybrid panel (IMpRH) and we sequenced the entire open reading frames of CACNB4 and SPG4 without finding any differences between healthy and affected piglets. An anti-epileptic drug treatment with ethosuximide did not change the severity of the disease, and pigs with CPA did not exhibit the corticospinal tract axonal degeneration found in humans suffering from hereditary spastic paraplegia, which is associated with mutations in SPG4. For all these reasons, the hypothesis that CACNB4, CHRNA1, KCNJ3, SCN2A or SPG4 are identical with the CPA gene was rejected.     

Post-mortem changes in porcine M. longissimus studied by solid-state 13C cross-polarization magic-angle spinning nuclear magnetic resonance spectroscopy

Solid-state (13)C cross-polarization (CP) magic-angle spinning (MAS) nuclear magnetic resonance (NMR) experiments are carried out for the first time on rapidly frozen muscle biopsies taken in M. longissimus in vivo and at 1 min, 45 min, and 24 h post-mortem from three pigs. Two of the pigs were CO(2)-stunned (control animals), and one was pre-slaughter-stressed (treadmill exercise) followed by electrical stunning to induce difference in metabolism post-mortem. (13)C resonance signals from saturated and unsaturated carbons in fatty acids, carboxylic carbons, and carbons in lactate and glycogen are identified in the solid-state NMR spectra. The (13)C CP MAS spectra obtained for post-mortem samples of the stressed, electrically stunned pig differ significantly from the post-mortem control samples, as the intensity of a resonance line appearing at 30 ppm, assigned to carbons of the methylene chains, is reduced for the stressed pig. This spectral difference is probably due to changes in lipid mobility and indicates altered membrane properties in the muscle of the stressed/electrically stunned animal when compared with the control animals already 1 min post-mortem. In addition, the post-mortem period changes in glycogen carbons can be estimated from the (13)C CP MAS spectra, yielding a correlation of r = 0.74 to subsequent biochemical determination of the glycogen content.     

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.     

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.     

Favism-inducing toxins in broad beans (Vicia faba): Estimation of the vicine contents of broad bean and other legume samples

A method for the estimation of vicine in leguminous seeds and related material has been developed. It involves the extraction and isolation of mixed pyrimidine bases, thin layer chromatography and spectrophotometry. Results obtained withVicia faba (broad bean) samples from various sources are recorded and reference made to data obtained from some other leguminous crops.     

Synthesis of lipophilic clovamide derivatives and their antioxidative potential against lipid peroxidation

Some N-(hydroxycinnamoyl)-L-tyrosine and L-DOPA alkyl esters were synthesized and evaluated as a variation of the clovamide (N-caffeoyl-L-3,4-dihydroxyphenylalanine) structure, a known antioxidant found in red clover. The amides were prepared in good yields starting from methyl and dodecylesters of L-tyrosine and L-DOPA by reacting with the N-hydroxysuccinimidyl esters of ferulic, sinapic, and acetyl-protected caffeic acid, respectively. In the DPPH* (2,2-diphenyl-1-picrylhydrazyl) and superoxide radical quencher assays they showed radical scavenging activity equal to or higher than those of the standard antioxidants ascorbic acid and tocopherol. The antioxidative potentials of the clovamide derivatives against bulk lipid oxidation, as determined by the accelerated autoxidation of oils, were equal to or higher than those of the standard antioxidants; some of the compounds were able to protect an emulsion of linoleic acid/beta-carotene against oxidation. N-Caffeoyl L-tyrosine methyl ester and the N-cinnamoyl L-DOPA alkyl esters especially were potent antioxidants in bulk lipids and moderate protectants in emulsions.     

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).