DNA repair phenotype and dietary antioxidant supplementation.

Phytochemicals may protect cellular DNA by direct antioxidant effect or modulation of the DNA repair activity. We investigated the repair activity towards oxidised DNA in human mononuclear blood cells (MNBC) in two placebo-controlled antioxidant intervention studies as follows: (1) well-nourished subjects who ingested 600 g fruits and vegetables, or tablets containing the equivalent amount of vitamins and minerals, for 24 d; (2) poorly nourished male smokers who ingested 500 mg vitamin C/d as slow- or plain-release formulations together with 182 mg vitamin E/d for 4 weeks. The mean baseline levels of DNA repair incisions were 65.2 (95 % CI 60.4, 70.0) and 86.1 (95 % CI 76.2, 99.9) among the male smokers and well-nourished subjects, respectively. The male smokers also had high baseline levels of oxidised guanines in MNBC. After supplementation, only the male smokers supplemented with slow-release vitamin C tablets had increased DNA repair activity (27 (95 % CI 12, 41) % higher incision activity). These subjects also benefited from the supplementation by reduced levels of oxidised guanines in MNBC. In conclusion, nutritional status, DNA repair activity and DNA damage are linked, and beneficial effects of antioxidants might only be observed among poorly nourished subjects with high levels of oxidised DNA damage and low repair activity.

Mutation of SOD1 in ALS: a gain of a loss of function.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease caused by motoneuron loss. Some familial cases (fALS) are linked to mutations of superoxide dismutase type-1 (SOD1), an antioxidant enzyme whose activity is preserved in most mutant forms. Owing to the similarities in sporadic and fALS forms, mutant SOD1 animal and cellular models are a useful tool to study the disease. In transgenic mice expressing either wild-type (wt) human SOD1 or mutant G93A-SOD1, we found that wtSOD1 was present in cytoplasm and in nuclei of motoneurons, whereas mutant SOD1 was mainly cytoplasmic. Similar results were obtained in immortalized motoneurons (NSC34 cells) expressing either wtSOD1 or G93A-SOD1. Analyzing the proteasome activity, responsible for misfolded protein clearance, in the two subcellular compartments, we found proteasome impairment only in the cytoplasm. The effect of G93A-SOD1 exclusion from nuclei was then analyzed after oxidative stress. Cells expressing G93A-SOD1 showed a higher DNA damage compared with those expressing wtSOD1, possibly because of a loss of nuclear protection. The toxicity of mutant SOD1 might, therefore, arise from an initial misfolding (gain of function) reducing nuclear protection from the active enzyme (loss of function in the nuclei), a process that may be involved in ALS pathogenesis.

Orange juice vs vitamin C: effect on hydrogen peroxide-induced DNA damage in mononuclear blood cells.

The intake of fruits rich in vitamin C seems to increase the antioxidant defence of the organism. However, it is still not clear whether vitamin C alone is responsible for this effect. The aim of the present investigation was to study the effect of the intake of a single portion of blood orange juice (BOJ, 300 ml, providing 150 mg vitamin C) on mononuclear blood cell (MNBC) DNA damage, compared with a drink supplemented with the same amount of vitamin C (C-drink) or sugars (S-drink). Seven young healthy subjects were randomised in a repeated-measures design in which they received each drink on different occasions, 2 weeks apart. Blood samples were collected at baseline, every hour for 8 h, and at 24 h after the intake of each drink. Vitamin C was analysed at each time point by HPLC, whereas H2O2-induced MNBC DNA damage was evaluated at 0, 3 and 24 h by means of the comet assay. Plasma vitamin C concentration increased similarly following BOJ or C-drink intake and was not affected by the S-drink. DNA damage significantly decreased 3 h after BOJ intake (about 18 %; P < 0.01) and remained constant at 24 h (about 16 %; P < 0.01). No effect of the C-drink and S-drink was observed. In conclusion, the intake of a single portion of BOJ provided an early protection of MNBC against oxidative DNA damage; however, the protective effect of BOJ was not explained by vitamin C alone, thus other phytochemicals could be involved.

Flavanone plasma pharmacokinetics from blood orange juice in human subjects.

Some blood orange juice (BOJ) flavanones may have antioxidant, anti-inflammatory, anti-allergic, hypolipidaemic, vasoprotective and anticarcinogenic properties. The aim of the present study was to evaluate the pharmacokinetics of hesperetin and naringenin in human subjects after BOJ intake. In a cross-over study, seven healthy female volunteers consumed 150 and 300 ml BOJ corresponding to about 51-102 mg hesperetin and to 6-12 mg naringenin, respectively. Plasma samples were collected before, each hour for 8 h and 24 h after BOJ administration and analysed for their content of hesperetin and naringenin by liquid chromatography-MS/MS. The plasma concentrations of these compounds were dose dependent and the peak concentration (Cmax) was reached in 5.1 (sd 0.6) h after BOJ intake. The Cmax of hesperetin was 43.4 (sd 32.4) and 79.8 (sd 60.1) ng/ml after 150 and 300 ml intake, respectively. For naringenin the plasma peak was 16.4 (sd 11.9) and 34.0 (sd 20.6) ng/ml. Moreover, the conjugated forms of these flavanones represent more than 95 % of the plasma concentration. The results indicate that both hesperetin and naringenin are bioavailable after BOJ intake; naringenin seemingly more so than hesperetin.

Effect of a tomato-based drink on markers of inflammation, immunomodulation, and oxidative stress.

Regular consumption of tomato and its products is being consistently associated with lower risk of several types of cancer and, to a lesser extent, coronary heart disease. Among the many tomato components credited with healthful properties, carotenoids and particularly lycopene are being actively investigated. Given the recognized role of immune/inflammatory processes in atherogenesis, the effects of a tomato-based drink (Lyc-o-Mato), which was previously shown to afford DNA protection from oxidative stress, on the modulation of immune and inflammatory markers (by enzyme immunoessay), on basal lymphocyte DNA damage (by comet assay), and on F2-isoprostane excretion (by LC-MS/MS), were investigated in 26 healthy young volunteers. In a placebo-controlled, double-blind, crossover study, Lyc-o-Mato (5.7 mg of lycopene, 3.7 mg of phytoene, 2.7 mg of phytofluene, 1 mg of beta-carotene, and 1.8 mg of alpha-tocopherol) or a placebo drink (same taste and flavor, but devoid of active compounds) were given for 26 days, separated by a wash-out period. During the study subjects maintained their habitual, hence unrestricted, diet. TNF-alpha production by whole blood was 34.4% lower after 26 days of drink consumption, whereas the other parameters were not significantly modified by the treatment. In turn, modest effects of the regular intake of a tomato drink, providing small amounts of carotenoids, were found on the production of inflammatory mediators, such as TNF-alpha, in young healthy volunteers. Future intervention trials in subjects with low carotenoid status and/or compromised immune system will resolve the issue of whether carotenoids modulate immune parameters in humans.

Daily intake of a formulated tomato drink affects carotenoid plasma and lymphocyte concentrations and improves cellular antioxidant protection.

The salutary characteristics of the tomato are normally related to its content of carotenoids, especially lycopene, and other antioxidants. Our purpose was to verify whether the daily intake of a beverage prototype called Lyc-o-Mato((R)) containing a natural tomato extract (Lyc-o-Mato((R)) oleoresin 6 %) was able to modify plasma and lymphocyte carotenoid concentrations, particularly those of lycopene, phytoene, phytofluene and beta-carotene, and to evaluate whether this intake was sufficient to improve protection against DNA damage in lymphocytes. In a double-blind, cross-over study, twenty-six healthy subjects consumed 250 ml of the drink daily, providing about 6 mg lycopene, 4 mg phytoene, 3 mg phytofluene, 1 mg beta-carotene and 1.8 mg alpha-tocopherol, or a placebo drink. Treatments were separated by a wash-out period. Plasma and lymphocyte carotenoid and alpha-tocopherol concentrations were determined by HPLC, and DNA damage by the comet assay. After 26 d of consumption of the drink, plasma carotenoid levels increased significantly: concentrations of lycopene were 1.7-fold higher (P<0.0001); of phytofluene were 1.6-fold higher (P<0.0001); of phytoene were doubled (P<0.0005); of beta-carotene were 1.3-fold higher (P<0.05). Lymphocyte carotenoid concentrations also increased significantly: that of lycopene doubled (P<0.001); that of phytofluene was 1.8-fold higher (P<0.005); that of phytoene was 2.6-fold higher (P<0.005); that of beta-carotene was 1.5-fold higher (P<0.01). In contrast, the alpha-tocopherol concentration remained nearly constant. The intake of the tomato drink significantly reduced (by about 42 %) DNA damage (P<0.0001) in lymphocytes subjected to oxidative stress. In conclusion, the present study supports the fact that a low intake of carotenoids from tomato products improves cell antioxidant protection.