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This study examined the effect of the intake of orange juice provided freshly squeezed (FS) or processed using low-temperature pasteurisation (LP), high-pressure processing (HPP), or pulsed electric field (PEF) treatment on the serum carotenoid concentrations of 12 healthy individuals, aged 20–32 years, enrolled in a crossover study. Participants were instructed to consume 500 ml of orange juice/day for 14 days. Carotenoid concentrations in the orange juice as well as serum samples retrieved on days 7 and 14 were analysed via HPLC. A significant increase in serum xanthophyll concentrations, but not serum carotenes, was observed, with the highest increase in α- and β-cryptoxanthin. The processing technologies applied appeared to affect serum carotenoid concentrations, with concentrations being similar in the HPP and FS orange juice types. As high variability in serum carotenoid concentrations was observed, the effect of different technologies on serum carotenoid concentration warrants further studies with larger sample sizes., Begoña Olmedilla-Alonso participates in the Spanish Carotenoid Network (CaRed) funded by the Spanish MINECO (Ministry of Economy, Industry and Competitiveness) grant BIO2015-71703- REDT. Begoña Olmedilla-Alonso and M Pilar Cano participate in the European Carotenoid Network (EuroCaroten) funded by the European Commission COST Action CA15136. This work was supported by the Consejería de Educación, Comunidad Autónoma de Madrid, Spain (grants: CAM 07G/0040/2000 and 07G/0041/2000) and by the Instituto de Salud Carlos III, Spain (grant RCMN C03/08).

[Background]: The acquired resistance to chemotherapy represents the major limitation in the treatment of cancer. New strategies to solve this failure and improve patients' outcomes are necessary. The cancer preventive effect of β-cryptoxanthin has been widely described in population studies. Few reports support its putative use as an antitumoral compound. Here we focus on the therapeutic potential of β-cryptoxanthin individually or in combination with oxaliplatin in colon cancer and try to decipher the molecular basis underlying its effect. [Methods]: Apoptosis, viability and proliferation assays, mouse models, and an intervention study in 20 healthy subjects were performed. A PCR array was carried out to unravel the molecular putative basis of the β-cryptoxanthin effect, and further signaling experiments were conducted. Comet Assay was completed to evaluate the genotoxicity of the treatments. [Results]: β-Cryptoxanthin differentially regulates the expression of the P73 variants in vitro, in vivo, and in a human intervention study. This carotenoid decreases the proliferation of cancer cells and cooperates with oxaliplatin to induce apoptosis through the negative regulation of ΔNP73. The antitumoral concentrations of oxaliplatin decrease in the presence of β-cryptoxanthin to achieve same percentage of growth inhibition. The genotoxicity in peripheral blood mononuclear cells of mice decreased in the combined treatment. [Conclusions]: We propose a putative novel therapeutic strategy for the treatment of colon cancer based on the combination of β-cryptoxanthin and oxaliplatin. The combined regimen produced more benefit than either individual modality without increasing side effects. In addition, the concentration-limiting toxicity of oxaliplatin is reduced in the presence of the carotenoid., This study was supported by FIS-ISCIII grant PI11/00593 (cosupported with FEDER Funds), Fundacion Banco Santander, ISCIII-RTICC-RD12/0036/0041, Fundacón Científica AECC, and Comunidad de Madrid S2010/BMD-2344.

This paper reports a study of the physicochemical characteristics of lutein-enriched frankfurter-type products with two fat levels, and the lutein in vitro bioaccessibility in these products. In general, the high-fat products possessed better emulsion stability and greater hardness and chewiness than low-fat samples. For each fat level, the addition of lutein extract in olive oil reduced emulsion stability in meat products. The presence of lutein reduced the lightness and increased the redness of samples. Lutein was highly stable upon in vitro digestion, with overall recovery of over 84% at the end of the duodenal phase and very low isomerisation. Micellarisation was high but depended on the fat content, ranging from 29-34% (for the low-fat sample) up to 73-81% of the amount initially present (for the high-fat sample). Storage (22 days at 4 °C) did not significantly affect lutein content or bioaccessibility. Our results support the suitability of meat products as lutein carriers and as a means to increase the systematic intake of lutein. © 2009 Elsevier Ltd. All rights reserved.

Our aim was to assess the suitability of ultra-high performance liquid chromatography (UHPLC) for the simultaneous determination of biomarkers of vitamins A (retinol, retinyl esters), E (alpha- and gamma-tocopherol), D (25-OH-vitamin D), and the major carotenoids in human serum to be used in clinical practice. UHPLC analysis was performed on HSS T3 column (2.1 x 100 mm; 1.8 microm) using gradient elution and UV-VIS detection. The system allows the simultaneous determination of retinol, retinyl palmitate, 25-OH-vitamin D, alpha- and gamma-tocopherol, lutein plus zeaxanthin, alpha-carotene, beta-carotene, alpha- and beta-cryptoxanthin and lycopene. The method showed a good linearity over the physiological range with an adequate accuracy in samples from quality control programs. Suitability of the method in clinical practice was tested by analyzing samples (n = 286) from patients. In conclusion, UHPLC constitutes a reliable approach for nutrient/biomarker profiling allowing the rapid, simultaneous and low-cost determination of vitamins A, E, and D (including vitamers and ester forms) and the major carotenoids in clinical practice.

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To compare the in vitro bioaccessibility of lutein, zeaxanthin, β-cryptoxanthin, lycopene, and α-and β-carotenes from relevant dietary contributors, a gastrointestinal model was used to assess the stability, isomerization, carotenol ester hydrolysis, and micellarization. Salivar, gastric, duodenal, and micellar phases were extracted, with and without saponification, and analyzed by using a quality-controlled HPLC method. The stability of carotenoids under digestion conditions was >75%, regardless of the food analyzed, whereas micellarization ranged from 5 to 100%, depending on the carotenoid and the food. cis-Isomers were maintained in processed foods, but increased in fresh foods. Xanthophyll ester hydrolysis was incomplete (

[Aim of the study]: To assess the in vitro bioaccessibility of carotenoids, including xanthophyll esters, and tocopherols from fruits and vegetables., [Results]: Stability for carotenoids and tocopherols was over 70%. Xanthophyll esters were cleaved by cholesterol esterase but not by human pancreatic lipase. Less than 40% of the β-cryptoxanthin initially present was hydrolyzed and the amount of free xanthophylls recovered was higher when liquid was used than when fresh homogenized matrix was employed. cis-Isomers of β-carotene and lutein did not significantly increase during the process. Xanthophylls were more efficiently transferred into supernatants than tocopherols and β-carotene. cis-Carotenoids, epoxy-xanthophylls and ester forms were also transferred., [Conclusion]: The results are consistent with observations in other in vitro digestion models and human studies and support the usefulness of in vitro assessment to study food-related determinants of the bioavailability of carotenoids and tocopherols from fruits and vegetables., The support of the Ministerio de Ciencia y Tecnología (AGL 2002-04059-C02-01) and Instituto de Salud Carlos III (RCMN C03/08) is acknowledged.

[Objective]: To assess the carotenoid status in young type I diabetic patients and its relationship to the glycaemic control of the disease., [Design]: A follow-up study., [Setting]: Hospital Universitario Puerta de Hierro, Health Area VI of Madrid (Spain)., [Subjects]: Forty-seven type I diabetic patients, followed for 2.5 years., [Interventions]: Coinciding with physical examination and laboratory tests, serum levels of carotenoids were analysed by HPLC, and dietary intake of carotenoids was evaluated by a semiquantitative food frequency questionnaire and 3-day prospective dietary records., [Results]: In type I diabetic patients, average intake, serum levels and correlations between diet and serum levels of carotenoids were comparable to those in reference non-diabetic groups. Between-subjects seasonal variations were observed for β-cryptoxanthin intake and serum levels (higher in winter) and serum lycopene (higher in summer). Significant within-subjects seasonal changes were shown for dietary and serum β-cryptoxanthin and serum β-carotene. Serum carotenoids were unrelated to glycaemic control markers. Subjects with clinically acceptable glycaemic control showed lower lycopene intake than those with unacceptable control. Intake of carotenoids did not explain variance in insulin dose, fasting glycaemia, fructosamine or HbA1c. With the exception of lycopene, serum carotenoids were predicted by dietary intake, but in no case by fasting glycaemia, HbA1c or fructosamine., [Conclusion]: In type I diabetic patients, serum carotenoid concentrations and their variance are determined by dietary intake patterns, and are unrelated to the glycaemic control of the disease, as assessed by biochemical markers., The funding of Instituto de Salud Carlos III (RCMN C03/08) and Lilly, SA (Madrid, Spain) is acknowledged.

Introduccion y objetivos. El objetivo de este trabajo fueanalizar el dano por isquemia-reperfusion mediado por radicales libres que se produce durante el trasplante cardiaco y eva-luar el posible efecto citoprotector de la trimetazidina (TMZ). Material y metodo. Se realizaron 21 trasplantes cardiacos ortotopicos en cerdos. Dividimos los experimentos en 2 grupos: A (n = 11), en el que se realizo una proteccion miocardica estandar, y B (n = 10), en el que se administro TMZ en la cardioplejia empleada para parar el corazon donante (TMZ, 10?5 mol/l), como pretratamiento intravenosodel receptor (TMZ, 2,5 mg/kg) y como parte de la cardio-plejia infundida en el receptor antes de despinzar la aorta(TMZ, 10?5 mol/l). Se tomaron muestras de sangre del senocoronario del receptor en 3 momentos: basal, isquemia y reperfusion. Se determino la concentracion de malonildial-dehido como marcador de peroxidacion lipidica y de variosantioxidantes: glutation peroxidasa, glutation reductasa,superoxido dismutasa, a-tocoferol, retinol y estado de antioxidantes totales. Resultados. Durante la isquemia-reperfusion aumentola produccion de malonildialdehido y la actividad de losantioxidantes enzimaticos, mientras que el retinol disminuyo. El incremento de malonildialdehido y de la actividad de la glutation peroxidasa entre el momento basal y la reperfusion fue significativamente mayor en el grupo A. Conclusiones. Durante el trasplante se incremento progresivamente el nivel de peroxidacion lipidica y se activaronlos sistemas antioxidantes intracelulares. La TMZ ejercio un efecto citoprotector y limito el dano por isquemia-reperfusion generado por los radicales libres, ademas de modificar el patron de reaccion de parte de los sistemas de defensa.

10 páginas., The bioavailability of vitamin C from pulsed electric fields (PEF)-treated vegetable soup in comparison with freshly made (FM) vegetable soup—gazpacho—and its impact on 8-epiPGF2α and uric acid concentrations in a human population were assessed. For this purpose six subjects consumed 500 ml PEF-treated vegetable soup/day, and six subjects consumed 500 ml FM vegetable soup/day for 14 days. On the first day of the study, the subjects drank the vegetable soup in one dose (dose–response study), and on days 2–14 they consumed 250 ml in the morning and 250 ml in the afternoon (multiple-dose–response study). Blood was collected every hour for 6 h on the first day and again on days 7 and 14. All blood samples were analyzed for vitamin C, 8-epiPGF2α, and uric acid. The maximum increase in plasma vitamin C occurred 3 h post-dose in both the PEF and the FM groups. Vitamin C remained significantly higher (P≤0.05) on days 7 and 14. The plasma 8-epiPGF2α concentration was significantly lower at the end of the study in both the PEF group (P=0.002) and the FM group (P=0.05). Plasma levels of vitamin C and 8-epiPGF2α were inversely correlated in both groups (r= − 0.549, P=0.018; and r= − 0.743, P=0.0004, respectively). To summarize, drinking two servings (500 ml) of PEF-treated or FM gazpacho daily increases plasma vitamin C and significantly decreases 8-epiPGF2α concentrations in healthy humans.

Free radicals are involved in ischemia-reperfusion injury and inflammatory processes. The commercial formulation of the anesthetic propofol contains gamma-tocopherol and delta-tocopherol, which may exert antioxidant effects during transplantation. Animals were randomly assigned to a control group or experimental groups for lung transplantation after 3 and 24 h of ischemia. Individual tocopherols, malondialdehyde, biochemical indices, and hemodynamic, blood gas, and ventilatory parameters were determined during reperfusion. Results showed that administration of commercially available propofol provoked a time- and dose-dependent increment in serum gamma-tocopherol and delta-tocopherol in control animals and in the group receiving lungs subjected to 3 h of ischemia, but not in the group with 24 h of ischemia. Malondialdehyde levels increased during reperfusion and did not differ significantly between the two experimental groups, which did not differ with respect to lung function either. gamma-Tocopherol, supplied by the anesthetic, may act as an antioxidant that is consumed during reperfusion. This potential effect could be relevant to the choice of anesthetic agents in situations where free radical damage to tissues is expected.

Lutein is one of the most widely found carotenoids distributed in fruits and vegetables frequently consumed. Its presence in human tissues is entirely of dietary origin. Distribution of lutein among tissues is similar to other carotenoids but, along with zeaxanthin, they are found selectively at the centre of the retina, being usually referred to as macular pigments. Lutein has no provitamin A activity in man but it displays biological activities that have attracted great attention in relation to human health. Epidemiological studies have shown inconsistent associations between high intake or serum levels of lutein and lower risk for developing cardiovascular disease, several types of cancer, cataracts and age-related maculopathy. Also, lutein supplementation has provided both null and positive results on different biomarkers of oxidative stress although it is effective in increasing macular pigment concentration and in improving visual function in some, but not all, subjects with different eye pathologies. Overall, data suggest that whereas serum levels of lutein have, at present, no predictive, diagnostic or prognostic value in clinical practice, its determination may be very helpful in assessing compliance and efficacy of intervention as well as potential toxicity. In addition, available evidence suggests that a serum lutein concentration between 0·6 and 1·05 μmol/l seems to be a safe, dietary achievable and desirable target potentially associated with beneficial impact on visual function and, possibly, on the development of other chronic diseases. The use of lutein as a biomarker of exposure in clinical practice may provide some rationale for assessing its relationship with human health as well as its potential use within the context of evidence-based medicine.

Within Europe there are differences in cardiovascular disease (CVD) risk between countries and this might be related to dietary habits. Oxidative modification of LDL is suggested to increase the risk of CVD and both the fatty acid and antioxidant content of LDL can affect its oxidation. In the present study, concentration of LDL fatty acid and antioxidant micronutrients (tocopherols and carotenoids) andex vivooxidative resistance of LDL (lag phase) was compared in volunteers from five countries with different fruit and vegetable intakes and reported rates of CVD. Eighty volunteers (forty males, forty females per centre), age range 25–45 years, were recruited from France, Northern Ireland, UK, Republic of Ireland, The Netherlands, and Spain, and their LDL composition and lag phase were measured. There were some differences in LDL carotenoid and α-tocopherol concentrations between countries. α-Tocopherol was low and β- + γ-tocopherol were high (PPP

Phytosterol (PS) intake may be used for hypercholesterolaemia in some groups although the presence of non-responders is well known. Carotenoids and PS/cholesterol may compete for the same transporters during absorption. As part of a randomized, double-blind, crossover, multiple-dose supplementation study with β-cryptoxanthin (β-Cx) and PS, single and combined, polymorphisms of ABCG8 (A632V) and NCPL1 (L272L) were determined in 19 post-menopausal women. Subjects carrying CC polymorphism for NCP1L1 (L272L) showed a net increase in total cholesterol and LDL after PS intake but, interestingly, displayed a decrease in both lipid fractions after consuming PS plus β-Cx. For the ABCG8 (A632V) gene, CT/TT carriers consuming PS also displayed an increase in total cholesterol and LDL, but this increment was much lower after the intake of PS plus β-Cx. Additionally, in CC carriers for ABCG8 (A632V), a greater decrease in total cholesterol and LDL was found after the intake of PS plus β-Cx compared to that observed after PS alone. Overall, our results suggest that β-Cx improves the response to PS in individuals carrying specific genetic polymorphisms (i.e. non-responders), opening the possibility to modulate the response to PS by food technology. (ClinicalTrials.gov NCT01074723).

Summary Background & aims Vitamin D deficiency is an important concern in clinical practice as hypovitaminosis D has been associated with important short and long-term health effects. The best indicator for vitamin D status is serum 25-OH-vitamin D but methodological issues limit the comparability between studies and the definition of cut-offs for hypovitaminosis D. Our aim was to assess the vitamin D status in outpatients as part of clinical practice. Methods We assessed, by a quality-controlled high-performance liquid chromatography (HPLC) method, the serum levels of 25-OH-vitamin D in 4040 outpatients belonging to Health Area VI of Madrid (Spain). Results Twenty-five percent of the subjects had levels 75 nmol/l) and 5% had “desirable” concentrations (>100 nmol/l). Serum distribution was not influenced by gender or season. Conclusions Prevalence of vitamin D insufficiency was high but depended on the cut-offs considered. Because of the health and economical consequences, cut-offs should be defined according to clinical (to avoid deficiency) and public health goals (optimal health promotion).

Carotenoids are thought to act as antioxidants in vivo, decreasing oxidative damage to biomolecules and thus protecting against coronary heart disease and cancer. However, human intervention studies with beta-carotene have given equivocal results in terms of cancer incidence. In an alternative molecular epidemiological approach, we have employed the 'comet assay' (single cell alkaline gel electrophoresis) to measure strand breaks, oxidized pyrimidines and altered purines in the DNA of lymphocytes from volunteers supplemented with alpha/beta-carotene, lutein, lycopene or placebo. In addition, we measured concentrations of the main serum carotenoids, and vitamins E and C, by HPLC. We report a significant negative correlation between basal concentrations of total serum carotenoids and oxidized pyrimidines. A similar correlation was seen between individual carotenoids (notably lutein and beta-carotene) and oxidized pyrimidines. However, carotenoid supplementation did not have a significant effect on endogenous oxidative damage. This suggests that there are some factors in the basal diet, probably found in fruit and vegetables, that decrease oxidative damage to DNA. In this case, basal serum carotenoids may simply be markers of consumption of fruit and vegetables, they themselves having little or no protective value.

Lutein, one of the major carotenoids present in serum, is also widely consumed by most populations. For the purpose of testing the potential health benefits of several carotenoids, lutein was supplied as part of an intervention trial to test whether the consumption of these food constituents reduces oxidative damage to human tissue components. Lutein from a natural source (15 mg/d as mixed ester forms) was supplied for 4 months to eighteen non-smoking, apparently healthy volunteers (nine men, nine women) aged 25–45 years. The serum carotenoid profile was analysed at baseline and monthly thereafter. On average, lutein concentrations increased 5-fold after the first month of supplementation (mean 1·34 (range 0·6–3·34) μmol/l). On reviewing the results, in those volunteers whose lutein levels surpassed 1·05 μmol/l (fourteeen of seventeen), we tentatively identified lutein monopalmitate along with another unidentified ester (possibly from a monoketocarotenoid) in serum. Lutein levels returned to baseline values and ester forms were not present 3 months after supplementation was discontinued. Their concentrations did not correlate with, and represented less than 3% of, lutein levels achieved in serum. They were observed before development of, and despite the presence of, carotenodermia. To our knowledge, this is the first time xanthophyll esters have been described in human serum. In view of the fact that xanthophyll esters have not been previously reported in serum and chylomicrons, it seems unlikely that these ester forms would be a reflection of the contents of the capsule. They may indicate a ‘ceiling effect’ on or saturation of the transport capacity for xanthophylls, and may have been re-esterifiedin vivobecause of the unusual dietary conditions. The determination of the physiological importance of this finding will require further investigation, although neither haematological nor biochemical changes were detected.

To establish reference ranges for use in clinical and epidemiological studies, we determined concentrations of retinol, α-tocopherol, β-carotene, α-carotene, β-cryptoxanthin, lutein, zeaxanthin, and lycopene in 450 Spanish control subjects and 123 Spanish patients with insulin-dependent diabetes mellitus (IDDM). Results were grouped according to sex, and samples were collected throughout the year. Concentrations of retinol were significantly lower and β-carotene and α-carotene were higher in women than in men, both in controls and IDDM subjects, whereas β-cryptoxanthin concentrations were higher only in control women. Conditional logistic regression analysis showed that retinol, β-carotene, and lycopene were the variables associated with diabetes. In comparison with other populations, our controls showed, in general, ordinary concentrations of retinol, comparatively low β-carotene and high β-cryptoxanthin concentrations, and a relatively high α-tocopherol/cholesterol ratio.

Intoxication from vitamin D supplements has been rarely reported, but nowadays, it occurs more frequently. The presence of the C-3 epimer of 25-hydroxyvitamin D(3) (3-epi-25-OH-D(3)) is highly prevalent in adults, although there is little information regarding its in vivo relevance, if any, especially under pathological conditions.Our aim was to assess the presence of the 3-epi-25-OH-D(3) in serum samples displaying 25-OH-D(3) concentrations indicative of hypervitaminosis D.A total of 58 samples displaying a wide range of concentrations of 25-OH-D(3) (64-439 ng/ml) by ultrafast liquid chromatography were consecutively recruited and reassessed for the presence of 3-epi-25-OH-D(3) using a second chromatographic system. Data from additional biochemical tests performed as part of the patient evaluation were also recorded.Mean relative contribution of 3-epi-25-OH-D(3) was less than 4%, and concentrations ranged from 2-28.6 ng/ml. Serum levels of the C3 epimer, but not the relative contribution, correlate with serum 25-OH-D(3). Overall, in subjects with 25-OH-D(3) concentrations indicative of hypervitaminosis D, the presence of the C-3 epimer and its levels were apparently unrelated to age, serum markers of renal and liver function, acute-phase reactants, and the presence of hypercalcemia. 3-Epi-25-OH-D(3) did not correlate with PTH, but subjects displaying PTH suppression (14 pg/ml) showed higher concentrations of 3-epi-25-OH-D(3).The relative contribution of 3-epi-25-D(3) was not significantly altered during hypervitaminosis D, although the absolute levels reached in serum may be biologically relevant. From a clinical viewpoint, although the small size of the group may affect the lack of relationships, the presence of 3-epi-25-OH-D(3) was apparently unrelated to serum markers of renal and liver function, acute-phase reactants, PTH, and the presence of hypercalcemia.

A Criminale-Ericksen-Filbey (CEF) constitutive equation was used to simulate the behaviour of HDPE samples of different molecular weights and blends with UHMWPE. Shear rate dependent viscosity and first normal stress coefficient data were obtained by capillary rheometry. The extruder operation point was calculated by using the central finite differences method and the result was compared with the experimental and power law values. At low temperatures, the CEF gave better approximations.

Carotenoids are one of the major food micronutrients in human diets and the overall objective of this review is to re-examine the role of carotenoids in human nutrition. We have emphasized the attention on the following carotenoids present in food and human tissues: β-carotene, β-cryptoxanthin, α-carotene, lycopene, lutein and zeaxanthin; we have reported the major food sources and dietary intake of these compounds. We have tried to summarize positive and negative effects of food processing, storage, cooking on carotenoid content and carotenoid bioavailability. In particular, we have evidenced the possibility to improve carotenoids bioavailability in accordance with changes and variations of technology procedures. © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

In vitro models based on human physiology have been developed as simple, inexpensive and reproducible tools to predict the bioavailability of different food components. Our aim was to assess the applicability of an in vitro digestion model to estimate the bioaccessibility of fat-soluble vitamins contained in commercially available liquid milk. For vitamin A, the rate and degree of hydrolysis of both endogenous and synthetic ester forms varied according to the type of milk (skimmed versus whole), although the amount of free retinol transferred into micelles was equivalent. For vitamin E, there was apparently no hydrolysis of tocopheryl acetate contained in fortified whole milk, while it was incomplete in the skimmed milk. Overall, in vitro results appear consistent with in vivo observations supporting the potential applicability and predictive value of the in vitro approach to assess the bioavailability of fat-soluble vitamins from dairy matrices., The support of the Ministerio de Ciencia y Tecnología (AGL 2002-04059-C02-01) and Instituto de Salud Carlos III (RCMN C03/08) is acknowledged.

Background: Dietary carotenoids have attracted a great deal of attention due to their potential clinical relevance in conditions such as age-related maculopathy, osteoporosis, cardiovascular disease, and cancer. Surgical procedures have become the primary treatment of severe obesity, although nutrient deficiencies are common and long-term metabolic sequelae remain unknown. Thus, our aim was to assess the carotenoid status in serum of subjects after obesity surgery. Methods: We evaluated the status of lutein, zeaxanthin, α- and β-cryptoxanthin, lycopene, α- and β-carotene, and fat-soluble vitamins by a quality-controlled high-performance liquid chromatography method in serum of 53 patients. Subjects were consecutively included as they were monitored for nutritional status after Roux-en-Y gastric bypass (RYGBP) or biliopancreatic diversion (BPD). Average follow-up time was 18 and 14 months for each protocol, respectively. Results: After obesity surgery, a consistent and continuous decline in all carotenoids to almost undetectable levels occurs, especially in those who underwent BPD diversion who, on average, displayed serum levels about one half to one third of those found in RYGBP patients. Conclusion: The hypocarotenemia observed after bariatric surgery may compromise the availability of carotenoids to tissues and the vitamin A status, reducing the fat-soluble antioxidant capacity and constituting an additional risk factor for several clinical conditions. Given the emerging role of carotenoids in disease prevention, dietary advice on carotenoid-rich and fortified foods or the use of supplements in these patients should be considered. © 2008 Springer Science + Business Media, LLC.

The food industry is playing an increasing role in the development and marketing of new products although little is known regarding the bioavailability of the phytochemicals they contain. Our aim was to assess the effect of the presence of absorption modifiers (milk and iron) on the in vitro bioaccessibility and the serum response in vivo of carotenoids and α-tocopherol from fruit juices. Thirty-two young women participated in a three-period (21 d each) supplementation study with a 2-week wash-out in between. Subjects consumed consecutively 2 × 250 ml/d vitamin C-fortified juices supplied as fruit juice, fruit juice containing milk and fruit juice containing milk and iron. Fasting blood samples were collected before and after each supplementation period. In vitro bioaccessibility of carotenoids and α-tocopherol was assessed by a static digestion model. Vitamin E and carotenoids from both studies were determined by HPLC. In vitro, xanthophyll ester hydrolysis and transference of free xanthophylls and α-tocopherol into the micellar phase were higher in the presence of absorption modifiers. In vivo, consumption of the fruit juices provoked significant increments (within-subject) of α-tocopherol and some carotenoids in serum. Dose-adjusted increments in serum of some carotenoids were higher when subjects consumed juices with milk and milk plus iron, although differences did not reach statistical significance. In conclusion, the presence of milk and milk plus iron do not influence the bioavailability of carotenoids and α-tocopherol from fruit juices in vivo. Our results support the use of in vitro models to assess food-related factors affecting bioavailability of carotenoids and tocopherols from foods.

7 páginas, 1 figura, 1 tabla., We assessed the bioavailability of lutein from lutein-fortified fermented milk using in vivo and in vitro approaches. Twenty-four volunteers were randomized to take lutein-fortified fermented milk at two levels of fortification. Single-dose bioavailability study (2×100 ml, ca. 8 or 16 mg of lutein) was performed using a three-point approach (baseline, 3.5 and 6.5 h). Multiple-dose study consisted of consuming one serving/day (ca. 4 or 8 mg/100 ml) for 14 days. Blood samples for biochemical, hematological and lutein analysis were drawn at baseline, Day 7 and Day 14. In vitro bioaccessibility was assessed by a static gastrointestinal digestion model. Lutein content, in vitro ester hydrolysis and micellarization, and lutein concentrations achieved in serum were analyzed by HPLC. In vivo, post-prandial response was higher using the high content fermented milk, but the percentage of absorption was not different according to the dose consumed. Net increments at Day 7 and Day 14 were significantly higher on consuming the high-dose milk as well. In vitro, lutein ester hydrolysis was incomplete regardless of the amount initially present. Free lutein released was higher using the high-dose fermented milk, but the percentage of hydrolysis was similar at both levels of fortification. In the micellar phase, the percentage of free and total lutein was not different according to the dose. Our results support the suitability of the fermented milk as a carrier of lutein esters and an in vivo dose-dependent effect upon regular consumption and suggest the usefulness of in vitro models to provide relevant information to predict in vivo responses., This work has been partially funded through a research agreement between Cognis Deutschland GmbH Co. and KG, and Fundación para la Investigación Biomédica Hospital Universitario Puerta de Hierro.

7 páginas, 4 tablas., [Aim of the study]: Ready-to-eat and pre-packed vegetables are increasingly accepted by consumers but little is known about the effect of these technological approaches on the bioavailability of the nutrients. To assess the effect of modified-atmosphere packaging (MAP) on the bioavailability in humans of carotenoids and tocopherols from broccoli. [Results]: Serum lutein increased significantly upon broccoli intake but those of β-carotene, α- and γ-tocopherol did not reach statistical significance. Serum changes were observed regardless of the type of broccoli consumed. [Conclusions]: Modified-atmosphere packaging does not affect significantly the in vivo bioavailability of carotenoids and tocopherols from broccoli, supporting its convenience for use by the food industry and consumers., This work was funded by Ministerio de Ciencia y Tecnología (AGL-2002-04059-C02-01) and the Instituto de Salud Carlos III (PI 051610), Spain.

Background: Diet and lifestyle are modifiable factors involved in the development and prevention of non-communicable diseases, including cardiovascular disease. Nut consumption, particularly walnut intake, has been inversely related to incident coronary heart disease (CHD) in observational studies and to improved lipid profiles in short-term feeding trials. Objective: To assess the potential functional effect associated with the regular consumption of walnut-enriched restructured meat products in subjects at risk for cardiovascular disease (CVD). Design: A crossover single-dose bioavailability study (n = 3) using γ-tocopherol as exposure marker and a crossover unblinded dietary intervention study (5 weeks) in subjects at risk (n = 25). Dietary intervention consisted of regular consumption of the meat product, with or without walnuts, five times per week for five weeks with a 1-month washout in between. Overnight fasting blood samples were collected on days 0, 12, 21, 28 and 35, coinciding with blood pressure and body weight recordings. Participants were asked to complete a diet record throughout the study. The functional effects were assessed using clinically relevant and related biomarkers of CHD: serum total, HDL and LDL cholesterol, triacylglycerols, homocysteine, vitamins B6 and B12, folic acid, α-tocopherol and platelet function test (obturation time). Results: The regular consumption of walnut-enriched meat products compared with that of the restructured meat products without added walnuts provokes a decrease in total cholesterol of 6.8 mg/dl (CI95%: -12.8, -0.85). Compared to baseline (mixed diet), meat products with walnuts decreased total cholesterol (-10.7 mg/dl, CI95%: -17.1, -4.2), LDL cholesterol (-7.6 mg/dl, CI95%: -2.2, -13.0) and body weight (-0.5 kg, CI95%: -0.1, -0.9) and increased γ-tocopherol (8.9 mg/dl, CI95%: 1.0, 16.8). Conclusions: The restructured meat products with added walnuts supplied in this study can be considered functional foods for subjects at high risk for CVD, as their regular consumption provokes a reduction in total cholesterol of 4.5% with respect to baseline values (mixed diet) and 3% with respect to the restructured meat without walnuts.

Summary: β-Cryptoxanthin displays a unique anabolic effect on bone calcification. In women with osteoporosis, serum β-cryptoxanthin and 25-OH-vitamin D3 showed a weak but significant correlation and exhibited a complementary seasonal distribution. The potential role of β-cryptoxanthin as a nutritional approach to improving bone health deserves further evaluation. Introduction: Dietary intake and serum levels of β-cryptoxanthin have been inversely related to different bone and joint disorders and in vitro and animal studies have shown that β-cryptoxanthin displays a unique anabolic effect on bone calcification. Due to the emerging role of β-cryptoxanthin in bone biology, we aimed to assess the serum distribution and variability of β-cryptoxanthin and their potential relation to 25-OH-vitamin D3 in women with osteoporosis. Methods: Serum concentrations of α- and β-cryptoxanthin and 25-OH- D 3 in women with osteoporosis (N=644) were analyzed using a quality-controlled high-performance liquid chromatography (HPLC) method. Results: Overall, significant seasonal variations were found for the three analytes and inter-individual variation was also high (60-73%). β-cryptoxanthin and 25-OH-vitamin D3 exhibited a marked complementary seasonal distribution in serum, with vitamin D displaying the highest values in summer and β-cryptoxanthin in winter. Conclusions: Given the anabolic effect of β-cryptoxanthin on bone calcification and its complementary seasonal distribution with respect to 25-OH-vitamin D3, the potential role of β-cryptoxanthin as a sustainable nutritional approach to improving bone health deserves to be further evaluated. © 2007 International Osteoporosis Foundation and National Osteoporosis Foundation., β-Cryptoxanthin . Bone health . human study . Osteoporosis. 25-OH-vitamin D3

A simple and sensitive isocratic reversed-phase high-performance liquid chromatography (HPLC) method for simultaneous determination of retinol, α-tocopherol and six carotenoids in human plasma was described. Sample preparation of the earlier published method was further developed by addition of ultrapure water, which enabled aqueous layer to freeze facilitating phase separation without pipetting thus also improving precision of the method. Developed method appeared to be less laborious and time consuming compared to the traditional extraction methods, which require removal of organic layer by pipetting. The recoveries (absolute and relative) were between 80% and 103%. The intra-assay CVs were 1.1–4.0% (normal level) and 3.3–9.0% (low level). Inter-assay CVs were 5.3–8.8%. Reference method for all these analytes was not available, but a comparison with another published method was carried out. The results of the comparison matched satisfactorily. The method is used routinely in our laboratory in a large population-based study., This study was supported by the University of Kuopio (J. Karppi), Yrjö Jahnsson Foundation (J. Karppi), Orion-Farmos Research Foundation (J. Karppi), Academy of Finland Grant # 114526 (T. Nurmi) and Grant # 45155 (K. Nyyssönen).

Objective: To assess changes in carotenoid intake based on the variations in the consumption of fresh fruit and vegetables in the Spanish population over the period 1964-2004. Design: Consumption data of fresh fruit and vegetables from Family Budget Surveys carried out in 1964, 1980, 1990 and 2004. Consumption data (g per person per day) accounted for >90% of fruit and vegetable consumption at each time point. Quality controlled high-performance liquid chromatography analysis of the carotenoid composition of Spanish fruit and vegetable was used. Subjects: Randomly selected, private households throughout Spain (20 800 households in 1964, 30 311 households in 1980, 21 155 households in 1990 and 6000 households in 2004). Twelve vegetables and 16 fruits representing 89-96% of the total consumption of fresh fruit and vegetables were used. Results: Individual consumption of fruit and vegetables has changed over this period, altering the total and individual intake of carotenoids. Total carotenoid intake increased from 2.5 mg per person per day in 1964 to 4.1 mg per person per day in 1990, with a decrease to 3.3 mg per person per day in 2004. These increments are due to an increase in lycopene, α- and β-carotene, while a decrease in lutein and zeaxanthin is observed during the last decade. A continuous and consistent decrease in the relative contribution of lutein in the diet is observed over the period studied. Conclusion: Although the consumption of fruit and vegetables is still consistent with a Mediterranean-type pattern, modifications in the consumption of individual fruits and vegetables have provoked changes in total and specific carotenoid intake with potential relevance in human health. © 2007 The Authors.

7 páginas, 1 figura, 2 tablas.-- Comunicación presentada al Experimental Biology 2004, celebrado en Washington DC (USA) en Abril de 2004., Consumption of fruits and vegetables is associated with a reduced risk of death from all causes including heart disease and stroke. In this work, the bioavailability of vitamin C from a Mediterranean vegetable soup (gazpacho) constituted mainly of tomato, pepper and cucumber, and its influence on plasma vitamin C, 8-epi-prostaglandin F2α (8-epi-PGF2α), prostaglandin E2 (PGE2), monocyte chemotactic protein-1 (MCP-1), and the cytokines/tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 concentrations in a healthy human population were assessed. Six men and six women consumed 500 ml of commercial gazpacho per day for 14 days, corresponding to an intake of 78 mg of ascorbic acid per day. There were no differences (P=.22) in baseline plasma vitamin C concentrations between the men and women. The maximum increase (P, Supported in part by the Coordinated Grant CAM 07G/0040/2000- CAM 07G/0041/2000 (Comunidad Autónoma de Madrid), the Grant AGL2002-04059-C02-02 (Ministry of Science and Technology, Madrid, Spain) (MPC); a Fulbright/Ministry of Education, Culture and Sports Award for Postdoctoral Research in the United States of America, Visiting Scholar Program, Commission for Cultural, Educational and Scientific Exchange between the United States of America and Spain, and a Ramón y Cajal Research Contract (Ministry of Science and Technology, Madrid, Spain) (CS-M).

[Objective]: To optimize an HPLC method for the simultaneous determination of vitamins A, E and 25-OH-vitamin D., [Methods]: Modifications include sample volume, internal standard, temperature, detection and injection volume., [Results and discussion]: Validity was supported by performance in interlaboratory exercises. Utility and versatility in clinical practice were assessed in outpatients and hospitalized subjects. The method was shown to be accurate and reliable for the simultaneous determination of vitamins A, E and 25-OH-vitamin D in serum throughout the entire range of clinically relevant concentrations., The support of the Instituto de Salud Carlos III (RCMN C03/08) is acknowledged.

Background: Dairy products fortified with vitamins and minerals represent a growing market that is of interest to those sectors of the population with unbalanced diets and increased needs. However, there is little information on the bioavailability of micronutrients in milk products at dietary intake levels. Aim of the study: To evaluate the bioavailability of vitamins A and E in whole milk and fortified whole and skimmed milk in control subjects., Methods: A single-dose bioavailability study was performed using three commercially available milks (unfortified whole milk and whole and skimmed milk fortified with vitamins A and E). Nineteen volunteers (10 women and 9 men) ingested 430 ml of each milk on different days. The contents of retinol and α-tocopherol provided in the milks and the retinyl esters and α-tocopherol in triglyceride-rich lipoprotein fractions (TRL) from plasma collected for 6.5 h postprandially were assayed using a quality-controlled HPLC method. The relative absorption of vitamins A and E from milks was calculated on the basis of area under the curve (AUC) versus time curve estimations, adjusted for plasma volume and expressed as percentage of the amount of nutrient provided. Results: The total amounts of retinol and α-tocopherol provided ranged between 0.48 and 4.15 μmol and 0.41 and 32.49 μmol, respectively. The AUC value of retinyl palmitate in TRL was higher for fortified whole milk than for the other two milks (unfortified whole and fortified skimmed milk). The percent relative absorption of vitamin A did not differ among the three types of milk. The AUC for α-tocopherol was no different after the ingestion of any of these milks. Conclusion: The mean percentage of retinol absorption was apparently similar for the three types of milk, regardless of the amount of fat ingested with each type of milk and the vitamin A provided. © 2006 Steinkopff Verlag.

Fruits and vegetables are the major sources of biologically active compounds, and carotenoids and tocopherols constitute important groups in human diets. Bioavailability is a critical feature in the assessment of the role of micronutrients in human health, and the approaches to this issue include in vitro and in vivo methods. Our aim was to evaluate the bioavailability of carotenoids and tocopherols present in broccoli and to compare in vitro and in vivo approaches. Fourteen apparently healthy volunteers consumed 200 g broccoli once a day for seven days. Blood samples were drawn at baseline and after intervention to determine changes in lutein, β-carotene, and Α- and γ-tocopherol as relevant phytochemicals provided with this vegetable. Broccoli also was subjected to simulated gastrointestinal digestion to assess changes related to preabsorptive processes. Analytes in serum and at each phase of the digestion were assayed by high-performance liquid chromatography. During the intervention, the amounts supplied dally ranged from 2.4 to 3.1 mg lutein, 1.4 to 1.8 mg β-carotene, 4.5 to 6.8 mg Α-tocopherol, and 0.8 to 1.8 mg γ-tocopherol. Significant changes in serum in both men and women were observed only for lutein, whereas for γ-tocopherol a significant change was detected in women. No changes were observed for Α-tocopherol, β-carotene, retinol, the Α-tocopherol-to-cholesterol ratio, or serum lipids. Using the in vitro model, more than 75% of lutein, β-carotene, γ-tocopherol, and Α-tocopherol remained at the duodenal phase, whereas Incorporation Into the supernatants accounted for, This work was supported by the Ministerio de Ciencia y Tecnologia (AGL 2002- 04059-C02-01) and the Instituto de Salud Carlos III (RCMN C03/08).

Meat and meat products are essential components of diets in developed countries and despite the convincing evidence that relate them to an increased risk for CVD, a growing consumption of meat products is foreseen. Epidemiological studies show that regular consumption of nuts, in general, and walnuts in particular, correlates inversely with myocardial infarction and ischaemic vascular disease. We assess the nutritional basis for and technological approach to the development of functional meat-based products potentially relevant in cardiovascular disease (CVD) risk reduction. Using the available strategies in the meat industry (reformulation processes) and a food-based approach, we address the design and development of restructured beef steak with added walnuts, potentially functional for CVD risk reduction. Its adequacy as a vehicle for active nutrients is confirmed by a pharmacokinetic pilot study in humans using γ-tocopherol as an exposure biomarker in chylomicrons during the post-prandial state. Effect and potential “functionality” is being assessed by a dietary intervention study in subjects at risk and markers and indicators related to CVD are being evaluated. Within the conceptual framework of evidence-based medicine, development of meat-based functional products may become a useful approach for specific applications, with a potential market and health benefits of great importance at a population level., This work has been performed with financial support from the Ministerio de Ciencia y Tecnología (AGL 2001-2398-C03-02) and Red de Investigación Cooperativa del Instituto de Salud Carlos III, Spain (RCMN C03/08).

7 páginas, 1 figura, 2 tablas., Orange juice, a rich source of vitamin C, accounts for 60% of all fruit juices and juice-based drinks consumed in western Europe. Orange juice preservation is currently accomplished by traditional pasteurization. Pulsed electric fields (PEF) have been studied as a nonthermal food preservation method. Food technology needs in the area of processing are driven by nutrition. Therefore, the objectives of this study were to assess the bioavailability of vitamin C from pulsed electric fields–treated orange juice in comparison with freshly squeezed orange juice and its impact on 8-epiPGF2α concentrations (biomarker of lipid peroxidation) in a healthy human population. Six subjects consumed 500 mL/day of pulsed electric fields–treated orange juice and six subjects consumed 500 mL/day of freshly squeezed orange juice for 14 days, corresponding to an intake of about 185 mg/day of ascorbic acid. On the first day of the study, subjects drank the juice in one dose, and on days 2–14 they consumed 250 mL in the morning and 250 mL in the afternoon. Blood was collected every hour for 6 hours on the first day and again on days 7 and 14. In the dose-response study, the maximum increase in plasma vitamin C occurred 4 hours postdose. Vitamin C remained significantly higher on days 7 and 14 in both orange juice groups. Plasma 8-epiPGF2α concentrations was lower at the end of the study (P < 0.001) in both groups. Plasma levels of vitamin C and 8-epiPGF2α were inversely correlated. Pulsed electric fields–preservation of orange juice retains the vitamin C bioavailability and antioxidant properties of fresh juice with a longer shelf-life., This work was supported in part by the Coordinated Grant CAM 07G/0040/2000-CAM 07G/0041/2000 from the Comunidad Autónoma de Madrid; Grant AGL2002-04059- C02-02 from the Ministry of Science and Technology, Madrid, Spain (MPC); a Fulbright/Ministry of Education, Culture and Sports Award for Postdoctoral Research in the United States of America, Visiting Scholar Program, Commission for Cultural, Educational and Scientific Exchange between the United States of America and Spain (CS-M); and a Ramón y Cajal Research Contract from the Ministry of Science and Technology, Madrid, Spain (CS-M).

Introduction We need valid food frequency questionnaires (FFQ) to assess diet in preschool children using parental report. Methods We collected dietary information for 171 children aged 4–5 years from parents of the INMA study in Valencia, a prospective mother-child cohort study. A 105 food items FFQ was used to assess the child9s diet of previous year, at baseline and, on average, 10 months later. As the reference method, we estimated several carotenoids, vitamin C and E in the plasma of the children. Pearson (and Spearman) correlations were calculated for reproducibility (FFQ1 vs FFQ1), and validity by comparing nutrient estimates from FFQ to nutrient biomarkers (biochemical calibration). Results The average of correlation coefficients for reproducibility (Spearman r) between the two FFQ was 0.44 (0.44 for energy, 0.41 for protein, 0.39 for carbohydrate, 0.41 fat, 0.55 for β-carotene, 0.60 for vitamin C and 0.39 for vitamin E). The average of correlation coefficients for validity (Pearson r) between the mean of two FFQ and nutrients in plasma were 0.06 for α-tocopherol, 0.10 for lutein-zeaxanthin, 0.44 for β-cryptoxanthin (p Conclusions The FFQ showed a relatively good reproducibility and satisfactory agreements with most nutrient biomarkers measured in blood which may support its use as a valid instrument for dietary assessment in preschool children at these early ages.

A food frequency questionnaire (FFQ) and carotenoid database with information on α- and β-carotene, lutein, lycopene and β-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 β-carotene in all countries except Spain where spinach was most important. Likewise, carrots were also the main source of α-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, β-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. Chemicals/CAS: Anticarcinogenic Agents; Antioxidants; beta Carotene, 7235-40-7; Carotenoids, 36-88-4; cryptoxanthin, 472-70-8; Lutein, 127-40-2; lycopene, 502-65-8; Xanthophylls

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.

Lutein, one of the major carotenoids present in serum, is also widely consumed by most populations. For the purpose of testing the potential health benefits of several carotenoids, lutein was supplied as part of an intervention trial to test whether the consumption of these food constituents reduces oxidative damage to human tissue components. Lutein from a natural source (15 mg/d as mixed ester forms) was supplied for 4 months to eighteen non-smoking, apparently healthy volunteers (nine men, nine women) aged 25-45 years. The serum carotenoid profile was analysed at baseline and monthly thereafter. On average, lutein concentrations increased 5-fold after the first month of supplementation (mean 1.34 (range 0.6-3.34) mumol/l). On reviewing the results, in those volunteers whose lutein levels surpassed 1.05 mumol/l (fourteen of seventeen), we tentatively identified lutein monopalmitate along with another unidentified ester (possibly from a monoketocarotenoid) in serum. Lutein levels returned to baseline values and ester forms were not present 3 months after supplementation was discontinued. Their concentrations did not correlate with, and represented less than 3% of, lutein levels achieved in serum. They were observed before development of, and despite the presence of, carotenodermia. To our knowledge, this is the first time xanthophyll esters have been described in human serum. In view of the fact that xanthophyll esters have not been previously reported in serum and chylomicrons, it seems unlikely that these ester forms would be a reflection of the contents of the capsule. They may indicate a 'ceiling effect' on or saturation of the transport capacity for xanthophylls, and may have been re-esterified in vivo because of the unusual dietary conditions. The determination of the physiological importance of this finding will require further investigation, although neither haematological nor biochemical changes were detected.

Carotenoids are a group of fat soluble pigments which are present in the human being, both in blood, as in tissues, and which are obtained through the diet, mainly from fruits and vegetables. The interest of these compounds is due not only to the provitamin A activity of some of them, but also due to a whole series of biological activities such as: antioxidant or prooxidant, photo-protective, modulator of the immune response, anti-carcinogen, etc. The best analytical method available for the analysis of carotenoids is high performance liquid chromatography (HPLC), which is used in our study both for serum as for foods, and it is controlled throughout periodic quality controls. In this article we present the preliminary results of the levels of the major serum carotenoids (b-carotene, a-carotene, b-cryptoxanthin, lutein, zeaxanthin, and lycopene) in control subjects from five European countries, as well as indicating the major dietary contributors to the carotenoids intake in the Spanish population. The percentage of each carotenoid to the total of the carotenoids analyzed, varies according to the origin of the studied population. Ireland and the UK show a very similar carotenoids profile. France presents the highest levels of lutein and b-carotene, which are present simultaneously in green vegetables. Spain shows the lowest levels of b-carotene, along with the highest levels of b-cryptoxanthin, which in our country is supplied mainly by oranges and tangerines. The most abundant carotenoid in all countries was lycopene. The average daily intake of these carotenoids (from fresh fruits and vegetables) in our population, is 3.5 mg/day. Through the relationships between the dietary carotenoid contents and serum the identification of "biomarkers" have been proposed, which might be correlated with several pathological situations, and thus contribute to the prevention of certain diseases.

To establish reference ranges for use in clinical and epidemiological studies, we determined concentrations of retinol, alpha-tocopherol, beta-carotene, alpha-carotene, beta-cryptoxanthin, lutein, zeaxanthin, and lycopene in 450 Spanish control subjects and 123 Spanish patients with insulin-dependent diabetes mellitus (IDDM). Results were grouped according to sex, and samples were collected throughout the year. Concentrations of retinol were significantly lower and beta-carotene and alpha-carotene were higher in women than in men, both in controls and IDDM subjects, whereas beta-cryptoxanthin concentrations were higher only in control women. Conditional logistic regression analysis showed that retinol, beta-carotene, and lycopene were the variables associated with diabetes. In comparison with other populations, our controls showed, in general, ordinary concentrations of retinol, comparatively low beta-carotene and high beta-cryptoxanthin concentrations, and a relatively high alpha-tocopherol/ cholesterol ratio.

Dietary factors provide protection against several forms of DNA damage. Additionally, consumer demand for natural products favours the development of bioactive food ingredients with health benefits. Lutein is a promising biologically active component in the food industry. The EFSA Panel on Dietetic Products, Nutrition and Allergies considers that protection from oxidative damage may be a beneficial physiological effect but that a cause and effect relationship has not been established. Thus, our aim was to evaluate the safety and potential functional effect of a lutein-enriched milk product using the Comet Assay in order to analyze the baseline, the induced DNA-damage and the repair capacity in the lymphocytes of 10 healthy donors before and after the intake of the mentioned product. Our data suggest that the regular consumption of lutein-enriched fermented milk results in a significant increase in serum lutein levels and this change is associated with an improvement in the resistance of DNA to damage and the capacity of DNA repair in lymphocytes. Our results also support the lack of a genotoxic effect at the doses supplied as well as the absence of interactions and side effects on other nutritional and biochemicals markers.

An HPLC study of 18 of the fresh vegetables (raw and cooked) most frequently consumed in Spain was done to determine their carotenoid composition. The results are grouped according to the color of the edible portion of each: green, red-orange, or yellowish-white vegetables. beta-Carotene and lutein were found to be present in all of the vegetables analyzed except the sweet red pepper, which contains zeaxanthin but not lutein. Lutein and zeaxanthin are present in five of the vegetables analyzed, with the highest concentration of both of these components being found in spinach. In green and yellowish-white vegetables, lutein predominates over beta-carotene. Red-orange vegetables show a wider carotenoid profile, in which the lutein levels are surpassed by other carotenoids (e.g., lycopene in tomatoes, alpha- and beta-carotene in carrots). Boiling was not found to alter the carotenoid profile of the samples, but the amounts of carotenoids quantified were higher when compared to those in raw samples.

Carotenoids and retinol-equivalents in food composition tables from European countries (EPIC Study)

The isocratic separation of 14 carotenoids, as well as retinol, retinyl acetate, retinyl palmitate, α-tocopherol and tocopherol acetate, is accomplished in 12 minutes, using a Spheri-5-ODS column and acetonitrile:dichloromethane:methanol (70:20:10) as mobile phase, with two-channel, programmable multiwavelength detection. The carotenoids separated are as follows: lutein/zeaxanthin, canthaxanthin, β-apo-8′ carotenal, β-cryptoxanthin, echinenone, lycopene, γ-carotene, α-carotene, β-carotene, 9-cis-β-carotene, 15-cis-/13-cis-β-carotene, phytoene and phytofluene. The separation of lutein and zeaxanthin is obtained simply by changing the mobile phase to acetonitrile:methanol (85:15).

The methods described herein were developed for the quantitative analysis of 10 carotenoids (lutein, zeaxanthin, canthaxanthin, beta-apo-8′carotenal, beta-cryptoxanthin, echinenone, lycopene, gamma-carotene, alpha-carotene, beta-carotene/15-cis-beta-carotene), 3 retinoids (retinol, retinyl acetate, retinyl palmitate) and alpha-tocopherol and d-alpha-tocopherol acetate with two-channel detection and a single sample preparation. Two fast and simple methods are described using a Spheri-5-RP-18 column, with two different mobile phases-acetonitrile:dichloromethane:methanol (70:20:10) in 10 minutes and acetonitrile:methanol (85:15) in 25 minutes. These methods would be beneficial to food scientists, cancer researchers and epidemiologists for the accurate estimation of vitamin A activity and the antioxidant capability of different compounds present in the human diet and the levels of these compounds in human serum.

The results by Montonen et al. (1) regarding the potential role of dietary antioxidants in the prevention of type 2 diabetes are of considerable interest. The authors identify β-cryptoxanthin as a preventive factor, regardless of adjustment for other potential confounding factors, and point out that data support the hypothesis that “a sufficient intake of antioxidants plays a role in type 2 diabetes prevention.” Although the follow-up time and number of subjects are considerable, …