Jennifer Burns

Antioxidant flavonols from fruits, vegetables and beverages: measurements and bioavailability

Flavonols are polyphenolic secondary plant metabolites that are present in varying levels in commonly consumed fruits, vegetables and beverages. Flavonols have long held an interest for nutritionists, which has increased following a Dutch study in the early 1990s showing that dietary intake of flavonols was inversely correlated with the incidence of coronary heart disease. The main factors that have hindered workers in the field of flavonol research are (i) the accurate measurement of these compounds in foods and biological samples, and (ii) a dearth of information on their absorption and metabolism. This review aims to highlight the work of the authors in attempting to clarify the situation. The sensitive and selective HPLC procedure to identify and quantify common flavonols and their sugar conjugates is described. In addition, the results of an on-going screening program into the flavonol content of common produce and beverages are presented. The bioavailability of dietary flavonols is discussed with reference to an intervention study with onions, as well as pilot studies with tea, red wine and cherry tomatoes. It is concluded that flavonols are absorbable and accumulate in plasma and that consuming high flavonol-containing varieties of fruits and vegetables and particular types of beverages could increase their circulatory levels.

Daunorubicin-induced myocardial failure: Reversal by digoxin of an inability to use ATP for contraction

Abstract Daunorubicin (DNR) is an anti-cancer drug limited by fatal cardiac complications in which rhythm disturbances precede ventricular failure. We have used isolated working rat hearts to study the mechanism of myocardial DNR-induced work failure. DNR perfused at 19 to 38 μmol/l severely disturbed the aortic pressure profile and produced complete contractile failure within 90 min. It rapidly accumulates in isolated rat hearts to 2 μmol/g dry weight. Cardiac mechanical activity is not restored after removal of DNR from the perfusate. Nor is bound DNR eluted by DNR-free perfusion. DNR produces irreversible cardiac arrest with paradoxical conservation of ATP and creatine phosphate. Thus, DNR-induced work failure is associated with an inability to use ATP. Routes of ATP production appear functional. Normal production of lactate during perfusion indicates conservation of oxidative metabolism. At the end of the experimental period DNR-inhibited cells contain more lactate than controls. Myocardial fluid distribution is disturbed only at high perfusion concentrations of DNR. Total protection against DNR-induced myocardial work failure is provided by prior digitalization of the donor rats.

Amino acid transport and protein synthesis in energetically-stable calcium-tolerant isolated cardiac myocytes

Myocytes isolated by enzymic dispersion from adult rat ventricular tissue are shown to be energetically stable in the presence of 0.5 mM CaCl2. ATP and ADP content and rates of lactate production are comparable with those of intact myocardial tissue and consistent with these cells being tightly coupled. Addition of 2,4-dinitrophenol precipitates rapid changes in adenine nucleotide concentrations and a 10-fold increase in lactate production. Cardiac myocytes selectively transport neutral amino acids of the A and L classes. Transport of the amino acid analogue alpha-aminoisobutyric acid is an active, temperature-dependent and insulin-sensitive process. The apparent Km for alpha-aminoisobutyric acid transport is similar to that determined for embryonic cardiac cells. Mature myocytes incorporate labelled amino acids into cytoplasmic proteins with molecular weights ranging from 10 000 to 150 000. Newly synthesised protein is metabolically stable. The data establishes calcium-tolerant myocytes as an experimental system offering many advantages over whole hearts for short- and long-term studies of protein synthesis and catabolism.

Characterization of digoxin binding and daunorubicin uptake by isolated mature rat cardiac myocytes

Myocytes isolated from ventricular muscle of mature rat heart have been used for characterization of digoxin binding and to establish whether a relationship exists between digoxin binding and uptake of daunorubicin. High- and low-affinity digoxin binding sites have been identified; respectively, 0.9 +/- 0.1 X 10(7) sites/myocyte, Kd 70-77 nM and 7 +/- 2 X 10(7) sites/myocyte, Kd 1.4-1.7 microM. Myocytes accumulate daunorubicin to an intracellular concentration 30-40 times that in the medium. We find no evidence that saturation of digoxin binding sites alters daunorubicin uptake or that daunorubicin influences binding of digoxin. Alteration of sarcolemmal membrane properties is demonstrated by inhibition of amino acid transport reflected in protein synthesis rates. Calmodulin activation of phosphodiesterase appears insensitive to daunorubicin.