Daniela Martini

Anthocyanin absorption, metabolism, and distribution from a wild blueberry-enriched diet (Vaccinium angustifolium) is affected by diet duration in the Sprague-Dawley rat.

The effect of wild blueberry consumption on anthocyanin (ACNs) distribution and metabolism in Sprague-Dawley (SD) rats was investigated. Thirty-two rats were fed for 4 or 8 weeks with a control (C) or a wild blueberry-enriched diet (8%) (WB). Anthocyanin profile in plasma, urine, feces, brain, and liver was evaluated by LC-MS/MS, and significantly increased in urine and not in feces after 8 weeks on the WB diet compared to that in 4 weeks, but no anthocyanins were detected in plasma, liver, and brain samples either in the C or WB groups. Metabolites of ACNs were detected in the plasma, urine, feces, and tissues of both the C and WB groups, but the urinary excretion of hippuric acid increased significantly after 4 and 8 weeks of WB consumption. Thus, it seems that ACNs are metabolized by the intestinal microflora to respective phenyl-alkyl acids, which can be further metabolized to benzoic acid. In conclusion, ACNs are bioavailable in rats, and the extent of their metabolism and excretion is based on diet duration. Additionally, urinary hippuric acid content could represent a potential biomarker of ACNs absorption and metabolism in the SD rat under the present experimental conditions.

DNA damage and repair activity after broccoli intake in young healthy smokers

Cruciferous vegetables contain compounds with antioxidant properties (e.g. carotenoids, vitamin C and folates) and can alter the activity of xenobiotic metabolism (i.e. isothiocyanates). These constituents may be particularly important for subjects who are exposed to free radicals and genotoxic compounds, including smokers. The aim of the study was to evaluate the effect of broccoli intake on biomarkers of DNA damage and repair. Twenty-seven young healthy smokers consumed a portion of steamed broccoli (250 g/day) or a control diet for 10 days each within a crossover design with a washout period. Blood was collected before and after each period. The level of oxidatively damaged DNA lesions (formamidopyrimidine DNA glycosylase-sensitive sites), resistance to ex vivo H(2)O(2) treatment and repair of oxidised DNA lesions were measured in peripheral blood mononuclear cells (PBMCs). We also measured mRNA expression levels of repair and defence enzymes: 8-oxoguanine DNA glycosylase (OGG1), nucleoside diphosphate linked moiety X-type motif 1 (NUDT1) and heme oxygenase 1 (HO-1). After broccoli consumption, the level of oxidised DNA lesions decreased by 41% (95% confidence interval: 10%, 72%) and the resistance to H(2)O(2)-induced DNA strand breaks increased by 23% (95% CI: 13%, 34%). Following broccoli intake, a higher protection was observed in subjects with glutathione S-transferase (GST) M1-null genotype. The expression level and activity of repair enzymes was unaltered. In conclusion, broccoli intake was associated with increased protection against H(2)O(2)-induced DNA strand breaks and lower levels of oxidised DNA bases in PBMCs from smokers. This protective effect could be related to an overall improved antioxidant status.

Effect of Broccoli Intake on Markers Related to Oxidative Stress and Cancer Risk in Healthy Smokers and Nonsmokers

Cruciferous vegetables (CVs) have been widely studied for their anticarcinogenic properties. The aim of the present study was to evaluate the protective effect of broccoli intake in smokers and nonsmokers. Twenty young healthy males (10 smokers and 10 nonsmokers) were randomized in a cross-over design and received a portion of broccoli (200 g) or maintained a controlled diet for 10 days each. The two periods were separated by a wash out period (20 days). Blood samples were collected at 0, 10, 30, and 40 days and used for the evaluation of DNA damage, insulin-like growth factor-I (IGF-I) and histone deacetylase (HDAC). Ex vivo protection from H 2 O 2 -induced DNA damage and endogenous DNA damage were evaluated in lymphocytes by means of the comet assay. Strand breaks decreased significantly after the broccoli diet in smokers as well as in nonsmokers (−22.2%; P < 0.0001), whereas oxidized purines decreased significantly only in smokers (−51.0%; P < 0.0001). Broccoli intake did not modify HDAC activity and IGF-I serum levels. Our results strengthen the importance of consuming CVs to increase cell protection against DNA damage. Future investigation, with different amount of broccoli and/or different time of exposure, is needed to understand the lack of effect on HDAC activity and IGF-I levels.

Identification and Quantification of Soluble Free, Soluble Conjugated, and Insoluble Bound Phenolic Acids in Durum Wheat (Triticum turgidum L. var. durum) and Derived Products by RP-HPLC on a Semimicro Separation Scale

A straightforward semimicro separation scale RP-HPLC method was developed for the identification and quantification of phenolic acids (PAs) occurring as soluble free, soluble conjugated, and insoluble bound compounds, which were independently extracted from wholemeal of durum wheat and from its derived products coarse bran, semolina, and dried pasta. A narrow bore column and a semimicro photodiode array detector (PDA) cell, in conjunction with a single quadrupole mass spectrometer, equipped with an electrospray ionization source (ESI-MS), were employed. The method was validated in terms of linearity of calibration graphs, limits of detection, limits of quantification, repeatability, and accuracy, which was evaluated by a recovery study. In each sample (wholemeal, coarse bran, semolina, and dried pasta), the total amounts of the three different forms of PAs were in the order bound > conjugated > free, with bound PAs accounting for 61.0-83.6% of the total PAs. Ferulic acid was the most abundant PA in both soluble free and insoluble bound forms, whereas sinapic acid predominated in the conjugated ones. The highest PA content, calculated as the sum of total PAs quantified in the three forms, was found in coarse bran, followed by wholemeal, semolina, and dried pasta.

Coffee Consumption and Oxidative Stress: A Review of Human Intervention Studies

Research on the potential protective effects of coffee and its bioactives (caffeine, chlorogenic acids and diterpenes) against oxidative stress and related chronic disease risk has been increasing in the last years. The present review summarizes the main findings on the effect of coffee consumption on protection against lipid, protein and DNA damage, as well as on the modulation of antioxidant capacity and antioxidant enzymes in human studies. Twenty-six dietary intervention studies (involving acute and chronic coffee intake) have been considered. Overall, the results suggest that coffee consumption can increase glutathione levels and improve protection against DNA damage, especially following regular/repeated intake. On the contrary, the effects of coffee on plasma antioxidant capacity and antioxidant enzymes, as well as on protein and lipid damage, are unclear following both acute and chronic exposure. The high heterogeneity in terms of type of coffee, doses and duration of the studies, the lack of information on coffee and/or brew bioactive composition, as well as the choice of biomarkers and the methods used for their evaluation, may partially explain the variability observed among findings. More robust and well-controlled intervention studies are necessary for a thorough understanding of the effect of coffee on oxidative stress markers in humans.

Improvement of lymphocyte resistance against H2O2-induced DNA damage in Sprague–Dawley rats after eight weeks of a wild blueberry (Vaccinium angustifolium)-enriched diet

The effect of wild-blueberry consumption on the resistance of H(2)O(2)-induced DNA damage was evaluated in the Sprague-Dawley (SD) rat. Thirty-two, four week-old SD rats were randomly assigned to two groups of sixteen rats each and fed a control diet (C) or a wild-blueberry (WB) diet for four or eight weeks. Following the dietary treatment, plasma antioxidant capacity was assessed by the TRAP assay. Ex vivo protection from H(2)O(2)-induced DNA damage was evaluated in lymphocytes by means of the comet assay. No significant effect was detected in plasma antioxidant capacity at four and eight weeks in both dietary groups. In contrast, the level of DNA damage was significantly lower in rats fed the WB diet compared with those on the C diet after eight weeks (11.± 1.0% vs 17.2 ± 2.4% DNA in tail, p ≤ 0.05) but not after four weeks. In conclusion, while WB consumption did not affect plasma antioxidant activity, it improved lymphocyte protection against oxidative damage, but only after eight weeks.

Effects of Genotype and Environment on Phenolic Acids Content and Total Antioxidant Capacity in Durum Wheat

ABSTRACT In cereals, phenolic acid (PA) content and total antioxidant capacity (TAC) may have a wide range of variability, probably because of several factors influencing the occurrence of grain antioxidants, which include genotype, environment, and their possible interactions. However, only a few studies have investigated the influence of these factors on durum wheat. In the present study, we investigated the impact of the genetic and environmental factors on the profile and content of PAs occurring as soluble free, soluble conjugated, and insoluble bound compounds, as well as on the TAC level, in three genotypes of durum wheat grown in three different Italian agroclimatic areas during two crop years. The results show that genotype, environment, and crop year have highly significant effects on TAC levels and on PA contents. In particular, TAC and free PAs are most influenced by year, whereas conjugated and bound PAs are most influenced by environment × year and genotype, respectively. Therefore, it is ev...

Claimed effects, outcome variables and methods of measurement for health claims proposed under European Community Regulation 1924/2006 in the framework of protection against oxidative damage and cardiovascular health

BACKGROUND AND AIMS The high number of negative opinions from the European Food Safety Authority (EFSA) to the requests for authorization of health claims is largely due to the design of human intervention studies, including the inappropriate choice of outcome variables (OVs) and of their methods of measurement (MMs). The present manuscript reports the results of an investigation aimed to collect, collate and critically analyse the information in relation to claimed effects, OVs and MMs, in the context of protection against oxidative damage and cardiovascular health compliant with Regulation 1924/2006. METHODS AND RESULTS Claimed effects, OVs and the related MMs were collected from EFSA Guidance documents and applications for authorization of health claims under Articles 13.5 and 14. The OVs and their MMs were evaluated only if the claimed effect was sufficiently defined and was considered beneficial by EFSA. The collection, collation and critical analysis of the relevant scientific literature consisted in the definition of the keywords, the PubMed search strategies and the creation of databases of references. The critical analysis of the OVs and their MMs was performed on the basis of the literature review and was aimed at defining the appropriateness of OVs and MMs in the context of the specific claimed effects. CONCLUSIONS The information provided in this document could serve to EFSA for the development of further guidance on the scientific requirements for health claims, as well as to the stakeholders for the proper design of human intervention studies aimed to substantiate such health claims.

In Vitro Bioaccessibility of Phenolic Acids from a Commercial Aleurone-Enriched Bread Compared to a Whole Grain Bread

Wheat aleurone, due to its potentially higher bioaccessibility and bioavailability of micronutrients and phenolic acids, could represent a useful ingredient in the production of commonly consumed cereal-based food. The aim of the present study was to investigate the in vitro bioaccessibility of phenolic acids both from an aleurone-enriched bread and from a whole grain bread. The two bread samples were firstly characterized for the phenolic acid content. An in vitro digestion was then performed in order to evaluate the release of phenolic acids. The results obtained suggest that the bioaccessibility of the phenolic acids in the aleurone-enriched bread is higher than in the whole grain bread. These in vitro results suggest the potential use of aleurone in the production of foods, and this may represent an attractive possibility to vehicle nutritionally interesting components to consumers.

Lycopene absorption in humans after the intake of two different single-dose lycopene formulations.

Lycopene is the main carotenoid present in tomato and its derivatives. It can also be obtained from the fungus Blakeslea trispora and its use as novel food ingredient was recently approved. The aim of the present study was to investigate the absorption of lycopene, in humans, after the intake of a single dose providing 15mg of lycopene from tomato extract (oleoresin 6%) or B. trispora (oil suspension 6%). Twelve female subjects were enrolled and divided into two groups: group 1 was assigned to the sequence tomato lycopene/wash-out/B. trispora lycopene, whereas group 2 followed the sequence B. trispora lycopene/wash-out/tomato lycopene. The formulations were consumed early in the morning with 5mL of sunflower oil, 100g of bread and 150mL of water. Blood was collected before consumption and after 2, 4, 6, 8, 10, 12 and 24h. Plasma lycopene concentrations were determined by HPLC analysis. On the whole, statistical analysis of data did not demonstrate a different effect of the type of lycopene source on the carotenoid absorption. The maximum increase in plasma lycopene concentration was about 40nmol/L for both products at 10-12h (p<0.05) post-consumption and decreased to basal values at 24h. A transient higher increase in lycopene concentration at 4-6h (p<0.05) after tomato lycopene with respect to B. trispora lycopene intake was observed. In conclusion, the intake of a single dose of the two liposoluble lycopene formulations revealed a comparable, small, but significant increase in plasma lycopene concentrations.