Homeostasis of metabolites in Escherichia coli on transition from anaerobic to aerobic conditions and the transient secretion of pyruvate

We have developed a method for rapid quenching of samples taken from chemostat cultures of Escherichia coli that gives reproducible and reliable measurements of extracellular and intracellular metabolites by 1 H NMR and have applied it to study the major central metabolites during the transition from anaerobic to aerobic growth. Almost all metabolites showed a gradual change after perturbation with air, consistent with immediate inhibition of pyruvate formate-lyase, dilution of overflow metabolites and induction of aerobic enzymes. Surprisingly, although pyruvate showed almost no change in intracellular concentration, the extracellular concentration transiently increased. The absence of intracellular accumulation of pyruvate suggested that one or more glycolytic enzymes might relocate to the cell membrane. To test this hypothesis, chromosomal pyruvate kinase ( pykF ) was modified to express either PykF-green fluorescent protein or PykF-FLAG fusion proteins. Measurements showed that PykF-FLAG relocates to the cell membrane within 5 min of aeration and then slowly returns to the cytoplasm, suggesting that on aeration, PykF associates with the membrane to facilitate secretion of pyruvate to maintain constant intracellular levels.

Immunologic reactivity and metabolic activity of blood lymphocytes associated with pathogenic form of allergic rhinosinusitis

We had studied functional and metabolic activity of blood lymphocytes in allergic rhinosinusitis (ARS) - atopic ARS and polypous rhinosinusitis (PRS). ARS is characterized by the increased level of CD25+ and HLA-DR+-lymphocytes in peripheral blood, IgA concentration in blood serum and sIgA in nasal secretion. At the same time the deviation of immune response is directed towards Th2-lymphocytes and is accompanied by high content of IL-4 and IL-6 in blood serum and nasal secretion. The activity of metabolic enzymes in blood lymphocytes in ARS is caused by exasperation of plastic processes, intensification of lipoid catabolism and increase of the speed of both anaerobic and aerobic oxidation. There are the following specific features of PRS: high level of CD19+-cells, activation of Thl-lymphocytes and high level of those cytokines, which they produce (IFN-γ and IL-2 in blood serum and nasal secretion). At the same time intensification of aerobic enzymes and lipoid metabolism and decreasing of anaerobic processes and aminoacid metabolism in blood lymphocytes are in the progress.

Thermal acclimation in fish: conservative and labile properties of swimming muscle

Given the rapid thermal equilibration of most fish with their environment, thermal compensation of metabolic and contractile properties is essential for the maintenance of locomotory capacities over a wide range of temperatures. The response of fish swimming performance, contractile properties of isolated fibers, myosin ATPase activity, and metabolic systems for ATP generation to short- and long-term changes in temperature have received sufficient study to allow one to identify certain constrained and labile properties. Sustained swimming performance and its components generally have their optimal performance and lowest thermal sensitivity within the range of temperatures frequently encountered by the organism. These principles are particularly well established for isolated enzyme systems. Furthermore, swimming performance and most of its components demonstrate thermal compensation on the evolutionary time scale. Temperature acclimation also leads to compensatory responses which, while quite species-specific, consistently increase the capacity for sustained swimming at low temperatures. The position of the thermal optimum for locomotion in relation to the width of a species' tolerance limits aids in predicting the species' capacity for thermal compensation during acclimation. Goldfish (Carassius auratus) and common carp (Cyprinus carpio), which tolerate temperatures 25–30 °C below their optimum for locomotion, show thermal compensation in terms of contractile properties, myosin ATPase activity, the proportion of red fibers in their axial musculature, and the levels of aerobic enzymes in their musculature. By contrast, striped bass (Morone saxatilis) and chain pickerel (Esox niger), which have lower optimal temperatures for locomotion, only increase the proportion of red fibers and (or) the levels of aerobic enzymes with cold acclimation. Finally, lake whitefish (Coregonus clupeaformis), which have their optimal temperature for locomotion at 12 °C, show none of these responses. Given that when thermal compensation occurs, aerobic enzymes in red muscle generally increase, the capacity of red muscle to generate ATP seems more temperature sensitive than other metabolic or contractile properties. Whether this compensatory response serves to counteract the effect of temperature on diffusive exchange between mitochondria and the cytoplasm or its effect on the catalytic capacity of aerobic metabolism remains to be established.

Influence of exercise training on reactivity and contractility of arterial strips from hypertensive rats

To determine whether the lowered resting blood pressure values in hypertensive rats were associated with changes in vascular reactivity and/or contractility, helical strips were obtained from different arteries from exercise-trained (running and swimming) hypertensive rats and from running normotensive rats. Each subgroup contained nontrained controls for comparison. Changes in muscle aerobic enzymes, maximum O2 consumption, and body weight indicated that a training effect had occurred. When norepinephrine was added in a dose-response manner to the testing chamber containing helical strips from either the descending aorta, femoral artery, or renal artery, there were no significant differences in reactivity (the negative log dose-response curve) attributable to the training of hypertensive or normotensive groups. However, the trained hypertensive rats exhibited a trend for lower contractility values (dyn/mm2) in some but not all of the strips tested. From these results, we concluded that the lowered resting blood pressures associated with exercise training of hypertensive rats could not be explained by changes in vascular reactivity of their arterial strips.