Mridula Chopra

Oxygen free radicals and congestive heart failure.

Plasma lipid peroxides (malondialdehyde) and thiols were measured in 45 patients with congestive heart failure and 45 controls. Malondialdehyde concentrations were significantly higher in the patients with congestive heart failure (median 9.0 nmol/ml interquartile range (IQR) 7.9-10.2) than in the controls (median 7.7 nmol/ml (IQR 6.9-9.2)). Plasma thiols were significantly lower in congestive heart failure (median 420 mumol/l (IQR 379-480)) than in the controls (median 463 mumol/l (IQR 445-525)). There was a significant but weak negative correlation between malondialdehyde and left ventricular ejection fraction (r = -0.35) and a positive correlation between plasma thiols and left ventricular ejection fraction (r = 0.39). This study provides clinical support for experimental data indicating that free radicals may be important in heart failure. It also suggests that the degree of free radical production may be linked to the severity of the disease.

A European carotenoid database to assess carotenoid intakes and its use in a five-country comparative study

A food frequency questionnaire (FFQ) and carotenoid database with information on alpha- and beta-carotene, lutein, lycopene and beta-cryptoxanthin was prepared and used to compare the carotenoid intakes in five European countries: UK, Republic of Ireland, Spain, France and The Netherlands. Eighty, age- (25-45 years) and sex-matched volunteers were recruited in each of the five countries. A FFQ and carotenoid database was prepared of the most commonly consumed carotenoid rich foods in the participating countries and the information was used to calculate frequency and intake of carotenoid-rich foods. The median total carotenoid intake based on the sum of the five carotenoids, was significantly higher (P < 0.05) in France (16.1 mg/day) and lower in Spain (9.5 mg/day,) than the other countries, where the average intake was approximately 14 mg/day. Comparison of dietary source of carotenoids showed that carrots were the major source of beta-carotene in all countries except Spain where spinach was most important. Likewise, carrots were also the main source of alpha-carotene. Tomato or tomato products, were the major source of lycopene. Lutein was mainly obtained from peas in Republic of Ireland and the UK, however, spinach was found to be the major source in other countries. In all countries, beta-cryptoxanthin was primarily obtained from citrus fruit. Comparing the data with that from specific European country studies suggests that the FFQ and carotenoid database described in the present paper can be used for comparative dietary intake studies within Europe. The results show that within Europe there are differences in the specific intake of some carotenoids which are related to different foods consumed by people in different countries.

Serum concentrations of carotenoids and vitamins A, E, and C in control subjects from five European countries

High intakes of fruits and vegetables, or high circulating levels of their biomarkers (carotenoids, vitamins C and E), have been associated with a relatively low incidence of cardiovascular disease, cataract and cancer. Exposure to a high fruit and vegetable diet increases antioxidant concentrations in blood and body tissues, and potentially protects against oxidative damage to cells and tissues. This paper describes blood concentrations of carotenoids, tocopherols, ascorbic acid and retinol in well-defined groups of healthy, non-smokers, aged 25-45 years, 175 men and 174 women from five European countries (France, UK (Northern Ireland), Republic of Ireland, The Netherlands and Spain). Analysis was centralised and performed within 18 months. Within-gender, vitamin C showed no significant differences between centres. Females in France, Republic of Ireland and Spain had significantly higher plasma vitamin C concentrations than their male counterparts. Serum retinol and alpha-tocopherol levels were similar between centres, but gamma-tocopherol showed a great variability being the lowest in Spain and France, and the highest in The Netherlands. The provitamin A: non-provitamin A carotenoid ratio was similar among countries, whereas the xanthophylls (lutein, zeaxanthin, beta-cryptoxanthin) to carotenes (alpha-carotene, beta-carotene, lycopene) ratio was double in southern (Spain) compared to the northern areas (Northern Ireland and Republic of Ireland). Serum concentrations of lutein and zeaxanthin were highest in France and Spain; beta-cryptoxanthin was highest in Spain and The Netherlands; trans-lycopene tended to be highest in Irish males and lowest in Spanish males; alpha-carotene and beta-carotene were higher in the French volunteers. Due to the study design, the concentrations of carotenoids and vitamins A, C and E represent physiological ranges achievable by dietary means and may be considered as reference values in serum of healthy, non-smoking middle-aged subjects from five European countries. The results suggest that lutein (and zeaxanthin), beta-cryptoxanthin, total xanthophylls and gamma-tocopherol (and alpha- : gamma-tocopherol) may be important markers related to the healthy or protective effects of the Mediterranean-like diet.

Antioxidant effects of Angiotensin-Converting Enzyme (ACE) inhibitors: free radical and oxidant scavenging are sulfhydryl dependent, but lipid peroxidation is inhibited by both sulfhydryl- and nonsulfhydryl-containing ACE inhibitors

Summary: With an assay that generates free radicals (FR) through photooxidation of dianisidine sensitized by riboflavin, 4 × 10−5M captopril, epicaptopril (SQ 14,534, captoprils stereoisomer), zofenopril, and fentiapril [all sulfhydryl (-SH)-containing angiotensin-converting enzyme (ACE) inhibitors] were shown effective scavengers of nonsuperoxide free radicals whereas non-SH ACE inhibitors were not. Captopril was a more effective FR scavenger at pH 5.0 than at pH 7.5. Captopril (2 × 10−5M) also scavenged the other toxic oxygen species hydrogen peroxide and singlet oxygen and inhibited microsomal lipid peroxidation. Finally, captopril reduced the amount of superoxide anion-radical detected after neutrophils in whole blood were activated with zymosan, probably by inhibiting leukocyte superoxide production.

Consumption of tomato products with olive oil but not sunflower oil increases the antioxidant activity of plasma

Health benefits of lycopene from tomato products have been suggested to be related to its antioxidant activity. Dietary fat may influence the absorption and hence the plasma levels and antioxidant activity of lycopene. In the present study, we have compared the effect of consumption of tomato products with extra-virgin olive oil vs. tomato products plus sunflower oil on plasma lycopene and antioxidant levels. Results show that the oil composition does not affect the absorption of lycopene from tomato products because similar levels of plasma lycopene (mean +/- SD) were obtained on feeding tomatoes (providing approximately 46 mg lycopene/d) for 7 d with either olive oil (0.66 +/- 0.26 vs 1.20 +/- 0.20 micromol/l, p <.002) or sunflower oil (0.67 +/- 0.27 vs. 1.14 micromol/l, p <.001). However, consumption of tomato products with olive oil significantly raised the plasma antioxidant activity (FRAP) from 930 +/- 150 to 1118 +/- 184 micromol/l, p <.01) but no effect was observed when the sunflower oil was used. The change (supplementation minus start values) in FRAP following the consumption of tomato products with oil was significantly higher for olive oil (190 +/- 101) than for sunflower oil (-9.6 +/- 99, p <. 005). In conclusion, the results of the study show that consumption of tomato products with olive oil but not with sunflower oil improves the antioxidant activity of the plasma.

No Significant Effects of Lutein, Lycopene or β-Carotene Supplementation on Biological Markers of Oxidative Stress and LDL Oxidizability in Healthy Adult Subjects

Objective: The objective of this study was to determine the effect of individual carotenoid supplementation on biochemical indices of oxidative status in apparently healthy adult males. Methods:The study was a placebo controlled single blind study. Healthy male volunteers (n=175) were assigned to four groups. They received daily supplements of β-carotene (15 mg), lutein (15 mg), lycopene (15 mg) and placebo for three months. The effects of the supplementation on antioxidant status were monitored by plasma carotenoid, vitamin C and A levels, glutathione (GSH and GSSG) concentrations, protein SH groups, erythrocyte antioxidant enzyme activities (Cu-Zn SOD, Se-GSH-Px) and susceptibility of LDL to copper-induced oxidation. Results:β-carotene, lycopene and lutein supplementation led to significant plasma and LDL increases in each of these carotenoids, without modifications of other carotenoid levels in plasma or in LDL. The supplementation failed to enhance the resistance of LDL to oxidation or to modify the LDL polyunsaturated/saturated fatty acid ratio. Vitamin C, GSH, protein SH groups and antioxidant metalloenzyme activities were also unchanged. Conclusion: We did not observe beneficial or adverse effects of lutein, lycopene or β-carotene supplementation on biomarkers of oxidative stress. In apparently healthy subjects, carotenoid supplementation does not lead to significantly measurable improvement in antioxidant defenses.

Effect of increased fruit and vegetable intake on the susceptibility of lipoprotein to oxidation in smokers

Objective: To evaluate the effect of an increased dietary intake of fruit and vegetables on susceptibility of LDL to oxidation in smokers and nonsmokers. Design: A descriptive and prospective study. Setting: Joseph Fourier University, Grenoble. Subjects: Volunteers were age and sex matched in the smoking and nonsmoking groups and were recruited by announcement. Interventions: Increased intake of fruits and vegetables for two weeks providing 30u2005mg/day of carotenoids. Main outcome measures: Circulating levels of beta-carotene, lutein, lycopene, α-tocopherol; susceptibility of LDL to oxidation. Cu-Zn superoxide dismutase (Cu-Zn SOD), and Se glutathione peroxidase (Se-GSH-Px) activities and reduced (GSH) and oxidized (GSSG) glutathione. Results: At entry (week 0: W0) smokers exhibited a lower plasma carotene level but the plasma parameters of oxidative stress and LDL oxidizability were not different from nonsmokers. After two weeks of increased intake of fruits and vegetables the circulating levels of carotenoids increased in smokers 23% and 11% in nonsmokers. At the same time the resistance of LDL to oxidation increased by 14% in smokers (P≤0.05) and by 28% in nonsmokers (P≤0.025). The mean whole bloodGSH level was higher in smokers at entry but returned to a concentration similar to nonsmokers at the end of the study. Conclusion: This pilot study indicates that an increased, carotenoid rich food intake through its inhibitory effect on the susceptibility of LDL to oxidation may be an interesting approach to reduce the risk of atherosclerosis both in smokers and nonsmokers. Sponsorship: This research has been supported by the European Union: AAIR project (AIR2-CT93-0888, DG 12SSMA).

Antioxidants and lipoprotein metabolism.

Lipoprotein metabolism plays a significant role in the pathogenesis of atherosclerosis and risk of vascular disease. Elevated levels of LDL and low levels of HDL are linked to the risk of cardiovascular disease. It is now widely accepted that oxidative modification of LDL affects the metabolism of lipoproteins, leading to their increased accumulation in arterial intima. The present paper will examine some of the accumulating evidence from studies showing relationships between dietary intake of antioxidant nutrients, i.e. vitamins E and C, the carotenoids and the flavonoids, and their influence on lipoprotein metabolism. In particular, the present paper will review the available literature on the affect of antioxidant supplementation on lipoprotein oxidation, and hence metabolism of LDL in human subjects.

Copper supplementation in humans does not affect the susceptibility of low density lipoprotein to in vitro induced oxidation (FOODCUE project).

The oxidative modification of low-density lipoprotein cholesterol (LDL) has been implicated in the pathogenesis of atherosclerosis. Copper (Cu) is essential for antioxidant enzymes in vivo and animal studies show that Cu deficiency is accompanied by increased atherogenesis and LDL susceptibility to oxidation. Nevertheless, Cu has been proposed as a pro-oxidant in vivo and is routinely used to induce lipid peroxidation in vitro. Given the dual role of Cu as an in vivo antioxidant and an in vitro pro-oxidant, a multicenter European study (FOODCUE) was instigated to provide data on the biological effects of increased dietary Cu. Four centers, Northern Ireland (coordinator), England, Denmark, and France, using different experimental protocols, examined the effect of Cu supplementation (3 or 6 mg/d) on top of normal Cu dietary intakes or Cu-controlled diets (0.7/1.6/6.0 mg/d), on Cu-mediated and peroxynitrite-initiated LDL oxidation in apparently healthy volunteers. Each center coordinated its own supplementation regimen and all samples were subsequently transported to Northern Ireland where lipid peroxidation analysis was completed. The results from all centers showed that dietary Cu supplementation had no effect on Cu- or peroxynitrite-induced LDL susceptibility to oxidation. These data show that high intakes (up to 6 mg Cu) for extended periods do not promote LDL susceptibility to in vitro-induced oxidation.

Evidence for oxidative stress in unstable angina

Objective—To determine whether unstable angina, which is characterised by recurring episodes of myocardial ischaemia and reperfusion, is associated with oxidative stress (that is, where there is an imbalance between oxidants, such as free radicals, which are in excess and antioxidants). Design—Between group comparison of patients with unstable angina, stable angina, and healthy controls. Setting—The coronary care unit and cardiac investigation ward of a regional cardiology centre. Patients—Twenty five consecutive patients admitted to the coronary care unit with unstable angina. Twenty five consecutive patients admitted to the cardiac investigation ward (patients with stable angina undergoing coronary angiography) were used as control for the presence of [ill], drug treatment, and smoking habit. Thirty eight healthy controls (hospital staff and patients admitted for minor surgical procedures who were otherwise healthy) were also studied. Main outcome measures—Thiobarbituric acid related substances (TBARS) in plasma and plasma reduced thiol (PSH) as indicators of oxidative damage to lipids and proteins respectively were measured. Coronary angiography was performed in all patients with stable angina and roughly half of those with unstable angina. Results—Mean (SEM) plasma TBARS in unstable angina and stable angina were 9·95 (0·36) nmol/ml and 9·14 (0·28) nmol/ml respectively (p = 0·08). Mean plasma TBARS in healthy controls were 8·09 (0·21) nmol/ml (p < 0·05 compared with both angina groups). Mean (SEM) PSH concentration in unstable angina was 4·21 (9) nmol/ml and in stable angina was 4·85 (9) nmol/ml (p < 0·05). Mean PSH in healthy controls was 5·64 (8) nmol/ml (p < 0·001 compared with both angina groups). The extent of coronary artery disease, use of medication, and smoking habit were not significantly different between the angina groups. Conclusions—Biochemical indicators of oxidative stress are more abnormal in unstable than stable angina. This is in keeping with experimental evidence that episodes of ischaemia and reperfusion lead to generation of free radicals and toxic oxygen species and depression of endogenous antioxidant activity. The clinical significance of this finding remains to be determined, although, experimentally, free radicals and toxic oxygen species have adverse effects on myocardial contractile function, myocardial electrical stability, endothelial mediated vasodilatation, and coagulation.