Aberrant energy-linked reactions in mitochondria isolated from the livers of rats infected with the liver fluke Fasciola hepatica.

The respiratory properties of mitochondria isolated from the livers of rats infected with the parasite Fasciola hepatica were examined. Oligomycin-sensitive ATPase activity was also examined during the acute stage (2-4 weeks post-infection). At 2,4 and 6 weeks post-infection, mitochondrial respiration in vitro (supported by site I and site II substrates) was completely uncoupled. Limited respiratory control had returned by 11 weeks post-infection, but complete recovery was not observed even at 21 weeks post-infection. At 4 weeks post-infection, uncoupled respiration (from all three energy-conserving sites) was also markedly attenuated (to the greatest extent with NADH-linked substrate). Except for pyruvate-supported respiration, this attenuation was not apparent at any other stage of the infection. The attenuation of pyruvate-supported respiration declined, but was still present, at 6 weeks post-infection. In addition to these perturbations in mitochondrial respiratory properties, mitochondrial ATPase activity at 4 weeks post-infection was insensitive to oligomycin, indicating a change in the structural integrity of the ATPase complex.     

Comparison of the effect of hypoxia on the secretion of the atrial natriuretic factor in spontaneously hypertensive and normotensive rats.

The effect of short lasting hypoxia on blood pressure, plasma atrial natriuretic peptide level and number of specific atrial granules were studied in 26 male spontaneously hypertensive and 24 normotensive Wistar rats. A great difference occurred in ANP secretion between hypertensive and normotensive rats. In the hypertensive animals elevated plasma ANP concentration (130 +/- 27 pg/ml) and decreased granularity in the right atria (73 +/- 2) were found on the first day of hypoxia with a slight elevation in urinary sodium content versus normotensive controls. The blood pressure also decreased although not significantly (190 +/- 14 mm Hg). In Wistar rats increased plasma ANP (130 +/- 34 pg/ml) and decreased atrial granularity versus normotensive controls (72 +/- 10 in the left and 113 +/- 16 in the right atrium) were observed only on the third day of hypoxia without changes in blood pressure and natriuresis. In SHR the rapid but short timed ANP release might be of right atrial origin and probably the consequence of a continuous and perhaps increased secretion of the peptide in normoxic conditions too. In Wistar rats the plasma ANP elevation could be secondary due to the increased plasma level of different vasoactive hormones to hypoxia. In the altered effect of ANP in hypertensive and normotensive hypoxic animals, structural and functional changes in the vascular bed may play a role.     

Phosphoenolpyruvate carboxykinase from Fasciola hepatica

Behm C. A. and Bryant C. 1982. Phosphoenolpyruvate carboxykinase from Fasciola hepatica. International Journal for Parasitology12: 271–278. The kinetic properties of a partially purified preparation of phosphoenolpyruvate carboxykinase (PEPCK) from F. hepatica were examined. The pH optimum for the carboxylation reaction is 5.8–6.2. The enzyme is more active with Mn²⁺ than Mg²⁺ and the Mn²⁺ saturation curve was sigmoid. Apparent K_m values for the substrates GDP, IDP, PEP and HCO₃^? were determined and found to be in the same range as those reported for other helminths except that the enzyme is less sensitive to low PEP concentrations. GTP and ATP at 0.5 and 1.0 mM inhibit the enzyme; the GTP inhibition was greater in the presence of Mg²⁺ than Mn²⁺ and was competitive with GDP. It was concluded that the activity of PEPCK from F. hepatica is controlled by the concentration of reactants and the ambient pH, that the accumulation of GTP is a sensitive mechanism for inhibiting the carboxylation reaction and that PEPCK activity in the cytosol is likely to be favoured over that of pyruvate kinase except when pH is high and PEP concentration low.     

Regulatory properties of a partially purified preparation of pyruvate kinase from Fasciola hepatica

It possesses sigmoid kinetics with PEP; FBP activation changes the relationship to a rectangular hyperbola. The enzyme is inhibited by malate, which competes with PEP; FBP relieves the inhibition slightly. ATP and bicarbonate ions are also inhibitory at high concentrations. ATP inhibition is mixed-competitive with PEP; bicarbonate inhibition is non-competitive. It is suggested that pyruvate kinase may regulate both lactate and acetate production by moderating the size of the cytosolic pyruvate pool.     

ATP synthesis in a succinate decarboxylase system from Fasciola hepatica mitochondria

Köhler P. B.,Ryant C. and Behm Carolyn A. 1978. ATP synthesis in a succinate decarboxylase system from Fasciola hepatica mitochondria. International Journal for Parasitology8: 399–404. Succinate decarboxylation was measured by the formation of ¹⁴CO₂ from 1,4-¹⁴C-succinate in a particle free, dialysed mitochondrial extract from liver fluke. It has an absolute requirement for Mg²⁺ and CoA. ATP, ADP and inorganic phosphate are essential for optimal activity. Ap₅A, an inhibitor of adenylate kinase, and glutathione are also necessary. GTP supports decarboxylation as well as ATP, provided ADP is also present. The formation of CO₂ and propionate greatly exceeds the amount of ATP and CoA initially present in the reaction mixture. A net, substrate-level phosphorylation of ADP occurs, the amount of ATP formed being equivalent to the production of CO₂ or propionate. This system is inhibited in flukes incubated in vitro with mebendazole.It is concluded that ATP is required to spark the fermentation system when succinate is the initial substrate and intermediate substrates are absent; that the terminal step in propionate formation is catalysed by a transferase which transfers CoA from propionyl CoA to succinate; and that ATP formation is coupled to the decarboxylation of methylmalonyl-CoA. A reaction scheme is presented.     

Studies of regulatory metabolism in Moniezia expansa: the role of phosphoenolpyruvate carboxykinase.

Phosphoenolpyruvate carboxykinase (PEPCK) from M. expansa has been partially purified and its behaviour in a range of different assay conditions has been determined. Different PEPCK's were found in the cytosol and mitochondria. Some kinetic parameters for each are presented. Both enzymes are activated by Mn²⁺; cytosolic PEPCK is also activated by Mg²⁺. The enzymes have pH optima in the range 6·4–7·0. They do not differ with respect to their apparent affinities for inosine and guanosine diphosphates, but the latter allows higher maximal activity. Little activity is observed with adenosine diphosphate. Adenosine and inosine triphosphates exert weak inhibitory effects on the Mn²⁺ activated enzymes; a much strongsr inhibition is exerted on the cytosolic enzyme when activated by Mg²⁺. A number of non-nucleotide compounds were tested for possible inhibitory effects with no success. The forward and back reactions catalyzed by PEPCK proceed at similar rates, suggesting that the enzyme may be readily raversible in vivo.     

Massage and stretching reduce spinal reflex excitability without affecting twitch contractile properties

Both stretching and massage can increase range of motion. Whereas the stretching-induced increases in ROM have been attributed to changes in neural and muscle responses, there is no literature investigating the ROM mechanisms underlying the interaction of stretch and massage. The objective of this paper was to evaluate changes in neural and evoked muscle responses with two types of massage and static stretching. With this repeated measures design, 30 s of plantar flexors musculotendinous junction (MTJ) and tapotement (TAP) massage were implemented either with or without 1 min of concurrent stretching as well as a control condition. Measures included the soleus maximum H-reflex/M-wave (H/M) ratio, as well as electromechanical delay (EMD), and evoked contractile properties of the triceps surae. With the exception of EMD, massage and stretch did not significantly alter triceps surae evoked contractile properties. Massage with and without stretching decreased the soleus H/M ratio. Both TAP conditions provided greater H/M ratio depression than MTJ massage while the addition of stretch provided the greatest inhibition. Both massage types when combined with stretching increased the duration of the EMD. In conclusion, MTJ and TAP massage as well as stretching decreased spinal reflex excitability, with TAP providing the strongest suppression. While static stretching prolongs EMD, massage did not affect contractile properties.