The metabolism of [1,2-13C2]glucose and [U-13C4]3-hydroxybutyrate was studied in rat brain with in vivo and in vitro 13C NMR spectroscopy, taking advantage, in particular, of homonuclear 13C-13C spin coupling patterns. After infusion of [1,2-13C2]glucose or [U-13C4]3-hydroxybutyrate into rats, the uptake of the substrates in brain and their metabolism to [1-13C]bicarbonate could be detected with in vivo 13C NMR spectroscopy. At the end of the infusion experiment, methanol/HCl/HClO4 extracts of the brain tissue were further analysed by high resolution 13C NMR spectroscopy. The 13C spin coupling patterns revealed entirely different isotopomer distributions for the closely related cerebral metabolites glutamate, glutamine and 4-aminobutyric acid. A quantitative analysis of the 13C spectra demonstrated (i) the existence of two kinetically distinct pools of glutamate, (ii) a pronounced CO2 fixation via pyruvate carboxylase in the glial cells accounting for as much as 38% of the oxaloacetate synthesis in the glial tricarboxylic acid cycle, (iii) a cerebral pyruvate recycling system contributing maximally 17% of the pyruvate metabolism through the pyruvate dehydrogenase in neurons, and (iv) a predominant production of 4-aminobutyric acid from glutamate synthesized in the neurons. In addition, the labelling pattern of N-acetyl aspartate upon infusion of labelled glucose or 3-hydroxybutyrate provided insight into the synthesis of this compound in mammalian brain. While the acetyl moiety originates from the metabolic equivalent of the C-1-C-2 part of cerebral glutamate, the aspartyl moiety is not in direct contact with the intermediates of glycolysis or of the tricarboxylic acid cycles.

3-Hydroxybutyric Acid↗

PubMedDate not supplied

Conformational changes of the phosphatidylcholine headgroup due to membrane dehydration. A 2H-NMR study.

The influence of hydration on the orientation of the phosphocholine dipole in bilayer membranes was studied with nuclear magnetic resonance. The phosphocholine headgroup of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) was deuterated at the two methylene segments. Phosphorus-31 and deuterium nuclear magnetic resonance measurements were made as a function of hydration in the range of 10-70 wt.% H2O revealing a distinct change in the alignment of the phosphocholine headgroup. With decreasing hydration the N+ end of the phosphocholine head group dipole moves closer to the hydrocarbon layer. The conformational change induced by the loss of water molecules at the membrane surface is qualitatively similar to that observed upon addition of polyhydroxyl compounds.

Deuterium↗

PubMedDate not supplied

Domain formation induced by lipid-ion and lipid-peptide interactions.

High sensitivity titration calorimetry was performed for metal ions such as calcium and lanthanum and for different types of Alzheimer peptides. Ca2+ adsorbs to mixed phosphatidylcholine (PC)/phosphatidylglycerol (PG) membranes with an endothermic reaction enthalpy of delta H approximately +0.1 kcal/mol. La3+ binds to sonified PC vesicles with a reaction enthalpy of delta H approximately + 1.8 kcal/mol. The binding constants are of the order of 10 M-1 for Ca2+ and 4 x 10(3) M-1 for La3+. The role of lipids in the random coil<-->beta-sheet equilibrium of different types of Alzheimer model peptides was investigated with circular dichroism (CD) and high sensitivity titration calorimetry. Alzheimer peptide beta AP(1-40)OH and several fragments of this peptide undergo a concentration-dependent, co-operative random coil<-->beta-sheet transition in solution which can be described by a linear association model with a nucleation parameter sigma approximately 0.2-0.01 and a growth parameter s approximately 10(4) M-1. Addition of sonified lipid vesicles containing negatively charged lipids shifts the equilibrium towards the beta-sheet conformation. This can be explained by an aggregation phenomenon at the lipid/water interphase. The cationic peptides are attracted to the negatively charged membrane surface causing a local increase in peptide concentration. The high peptide concentration, together with the ordering of the peptide molecules on the membrane surface, facilitates beta-sheet formation, constituting the first experimental evidence for the induction of beta-sheet formation via the membrane surface. The binding of Alzheimer peptide fragments to the lipid membrane is accompanied by an exothermic heat of reaction with delta H in the range -2 - -8 kcal/mol.

Adsorption↗

PubMedDate not supplied

[Ischemic preconditioning improves post-ischemic function but not energy metabolism of skeletal muscles].

Ischemic preconditioning (IP) refers to a phenomenon whereby short periods of ischemia reduce tissue damage after a subsequent sustained ischemia. The effect of IP before tourniquet ischemia of the extremities has not yet been evaluated. We developed a rat model of skeletal muscle ischemia and measured the effect of IP on postischemic function and high-energy phosphate levels. IP consisted in three cycles of 10 min ischemia and 10 min reperfusion each. IP improved significantly skeletal muscle function after 3 hours of ischemia and 2 hours of reperfusion. High-energy phosphate levels, however, remained unchanged. This study shows a protective effect of IP in skeletal muscles. These results furthermore suggest that the protection of energy metabolism is not a mechanism of IP in this model. IP could be easily performed before surgery of the extremities under tourniquet ischemia. The protective effect on postischemic skeletal muscle has therefore to be further investigated.

Animals↗