Trypanocidal CoQ analogues: their effect on other mitochondrial systems

1. A comparative study of compounds which inhibit the respiration of the infective form of the protozoan parasite Trypanosoma brucei brucei, such as salicylhydroxamic acid, other substituted benzhydroxamic acid, esters of 2,3- and 3,4-dihydroxybenzoic acid and structurally related compounds, showed that they have a remarkable degree of selectivity for the trypanosome as compared to rat liver mitochondria even though they are putative CoQ analogues and both respiratory systems are dependent on CoQ. 2. The minimal inhibition of mammalian mitochondrial function could not be assigned to inhibition of ubiquinone function in these mitochondria. 3. CoQ-reducing mitochondrial dehydrogenases from rat liver, trypanosomes and skunk cabbage (Symplocarpus foetidus) were insensitive to these inhibitors. 4. The alternative oxidase of skunk cabbage mitochondria was sensitive to a spectrum of trypanosome respiration inhibitors suggesting a similarity to the oxidase of the trypanosome although differing degrees of sensitivity and differing responses to alterations in the molecular structure of the inhibitors indicate that the milieu of the active sites are dissimilar.     

Effects of 30 days of creatine ingestion in older men

In this investigation we evaluated the effects of oral creatine (Cr) supplementation on body composition, strength of the elbow flexors, and fatigue of the knee extensors in 20 males aged 60-82 years who were randomly administered Cr or placebo (P) in a double-blind fashion. Subjects ingested either 20 g of Cr or P for 10 days, followed by either 4 g of Cr or P, respectively, for 20 days. Tests were conducted pre-supplementation and following 10 and 30 days of supplementation. Leg fatigue was determined using an isokinetic dynamometer; subjects performed 5 sets of 30 maximal voluntary contractions at 180 degrees x s(-1), with 1 min of recovery between sets. The strength of the elbow flexors was assessed using a modified preacher bench attached to a strain gauge. There was a significant interaction (P < 0.05; group x time) in leg fatigue following supplementation. However, this interaction appears to have resulted from a combination of the improved fatigue score by the Cr-supplemented group and the decreased fatigue score by the P-supplemented group, because when the simple main effects were analyzed for the groups individually, there was no significant difference over time for either of the groups. There were no significant differences in body mass, body density, or fat-free mass as assessed by hydrostatic weighing, or strength between the Cr-supplemented or P-supplemented groups. These data suggest that 30 days of Cr-supplementation may have a beneficial effect on reducing muscle fatigue in men over the age of 60 years, but it does not affect body composition or strength.     

Pilot-scale verification of a computer-based simulation for fractional protein precipitation

The development and experimental verification at pilot scale of a suite of models for the batch precipitation by two-cut ammonium sulphate salting-out of total protein and alcohol dehydrogenase from yeast homogenate is presented. The model consists of two elements: protein and enzyme solubility prediction and precipitate phase particle property prediction. An isotherm-based approach has been used successfully to describe solubility behaviour for a range of operating conditions typical of those obtained at a process scale. Estimation of the precipitate phase particle size distributions has been achieved through a discretized population balance approach using simplified terms to account for particle breakage and aggregation. The developed model accounts for the effects of average shear rate and residence time in the precipitation vessel across a two orders of magnitude range of scale. The size-density relationship for the precipitate phase has been defined. Results of simulations are compared with pilot scale verification data to confirm the validity of the models developed.     

Exercise-induced muscle damage, repair, and adaptation in humans

This study examined exercise-induced muscle damage, repair, and rapid adaptation. Eight college-age women performed three eccentric exercises of the forearm flexors. One arm performed 70 maximal contractions (70-MAX condition), and the other arm performed 24 maximal contractions (24-MAX) followed 2 wk later by 70 maximal contractions (70-MAX2). Criterion measures of serum creatine kinase, muscle soreness and pain, isometric strength, and muscle shortening were assessed before, immediately after, and for 5 days after each exercise. Significant changes in all criterion measures were found after the 70-MAX exercise with a slow recovery that was not complete by day 5 after exercise. The 24-MAX condition showed only small changes in the criterion measures. Changes in the criterion measures after the 70-MAX2 exercise were significantly smaller than those after the 70-MAX exercise. Results from this study, with regard to the ability of the muscle to adapt to exercise-induced damage, suggest that an adaptation takes place such that the muscle is more resistant to damage and any damage that does occur is repaired at a faster rate. It is also clear that a relatively small insult will produce this adaptation.     

INACCURACY OF ESTIMATIONS OF S PHASE FRACTION BY REDUCTION IN CLONING EFFICIENCY WITH HYDROXYUREA OR TRITIATED THYMIDINE

The current hypothesis, that the fractional reduction of cloning efficiency in semi-solid culture systems induced by pretreatment of the cells with hydroxyurea (HU) or [³H]TdR equals the fraction of cells initially in S phase, is tested. A lymphoblastoid cell line, SK-L7, with known cell cycle kinetics was exposed to cytotoxic concentrations of HU or suicidal doses of [³H]TdR and then initiated in semi-solid and liquid culture. Although approximately 0.6 of the initial population was in S, 1-hr exposures of HU at concentrations of up to 10^-2 M failed to reduce subsequent cloning efficiency. the 1-hr exposure to HU did not reduce either the immediate cell number or the gross population doubling rate over 24 hr. A 24-hr exposure to 10^-3 M HU reduced the cloning efficiency by approximately 98%, confirming the drug's cytotoxic capability. [³H]TdR at doses of 100 μCi/ml for 20–40 min reduced the cloning efficiency by approximately 60 and 70%, respectively. Although no cytotoxicity immediately after exposure was observed in either case, gross population doubling rate in liquid culture was reduced. While HU failed to reduce subsequent cloning efficiency, [³H]TdR reduced cloning efficiency by approximately the fraction of initial cells in S. the above hypothesis, therefore, cannot be applied naïvely as a technique for quantitating the fraction of a clonogenic cell population in S phase.