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Background C-reactive protein is an acute phase response marker and, in an epidemiological context, a predictor of cardiovascular risk. 25-Hydroxy-vitamin D is the best indicator for vitamin D status, but it can be altered by the presence of acute phase response. Our aim was to evaluate the association between serum concentrations of 25-hydroxy-vitamin D and C-reactive protein to assist the interpretation of vitamin D status in a clinical context. Methods We evaluated retrospectively 5076 patients ( n = 4087 women) assessed for 25-hydroxy-vitamin D and C-reactive protein simultaneously. Subjects were classified according to the origin as hospitalized patients ( n = 410) and outpatients ( n = 4666). Outpatients included patients from specialized ( n = 3943) and primary ( n = 723) care. Serum 25-hydroxy-vitamin D was determined by using liquid chromatography and serum C-reactive protein by using immunoturbidimetry. Results Concentrations of 25-hydroxy-vitamin D and C-reactive protein were significantly different between hospitalized subjects and outpatients but not for specialized and primary care settings. Serum concentrations of 25-hydroxy-vitamin D decreased as C-reactive protein increased. Hospitalized patients with C-reactive protein concentrations >30 mg/L showed a significant reduction of 25-hydroxy-vitamin D. In outpatients with C-reactive protein within the reference range (≤10 mg/L), C-reactive protein concentrations were not significantly different for serum 25-hydroxy-vitamin D concentrations >37.5 nmol/L. Conclusions Our data question the reliability and usefulness of assessing 25-hydroxy-vitamin D status as a biomarker of nutritional status in patients displaying acute phase response, especially at concentrations of C-reactive protein >30 mg/L. In addition, the present study shows that in subjects displaying C-reactive protein values within the reference range, serum concentrations of 25-hydroxy-vitamin D >37.5 nmol/L were not associated with lower concentrations of cardiovascular risk (as assessed by C-reactive protein concentrations).

The hypocholesterolemic effect and the modification of serum biomarkers of a dietary plant sterol (PS) intake, cholesterol precursors and cytokines after the consumption of milk-based fruit beverages with a milk fat globule membrane were evaluated by a randomized, double-blind, crossover, multiple dose bioavailability study. Postmenopausal women (n = 38) consumed daily 250 mL of a beverage with or without 2 g of PS added during 6 weeks in each of the study periods. With the intake of the PS-added beverage, significant decreases (mg dL-1) in serum total cholesterol (pre-treatment: 220.0 ± 27.8 vs. post-treatment: 212.9 ± 25.8; p < 0.05) and LDL-cholesterol (129.4 ± 28.5 vs. 121.7 ± 24.4; p < 0.05) were detected. The cholesterol precursor lathosterol (11.2%), markers of the dietary PS intake (campesterol 43.1% and β-sitosterol 32.5%), and anti-inflammatory IL-10 cytokine (22.5%) increased significantly, with a concomitant significant reduction in pro-inflammatory IL-1β (6.7%). No variations in HDL-cholesterol, other sterols (desmosterol and stigmasterol) or cytokines (IL-6, IL-8, IL-12p70 and TNF-α) were detected. These results indicated that this kind of PS-enriched milk-based fruit beverage is suitable during the period of clinical intervention, and its consumption may be an adequate way to improve PS functionality since a significant reduction in cholesterol levels has been observed. Therefore, the intake of this beverage could contribute to reduce the risk of cardiovascular disease also obtaining a beneficial effect on the serum inflammatory status in postmenopausal women.

[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.

Objective We aim to determine 3-epi-25-OH-D 3 in 155 leftover infant samples ( Methods Samples were randomly collected and assessed for 3-epi-25-OH-D 3 by HPLC. Biochemical data were recorded, if available, including C-reactive protein, calcium, liver and renal markers. Season of birth, gestational age (pre-term vs full-term), weight and height were recorded for infants Results 3-Epi-25-OH-D 3 was quantified in 97% of the samples. Serum levels of the epimer increased at the end of the pregnancy and decreased over the first year of life. 3-Epi-25-OH-D 3 was negatively associated with bilirubin and C-reactive protein but unrelated to gestational age, season, age after delivery, height and weight, and calcemia or renal function. Conclusions Based on the present data, both liver function and acute phase reactants should be considered when measuring 3-epi-25-OH-D 3 and assessing vitamin D status in infants.

Summary Background & aim Vitamin D deficiency is an important concern in clinical settings and recently, international agencies have recognised the importance of 25-OHD assays in defining vitamin D status. Thus, our aim was to assess the consistency of different vitamin D assays in clinical practice. Methods: 25-OH-vitamin D was measured in 332 patients by ultra-fast liquid chromatography (UHPLC) and two immunoassays (Liaison Total 25(OH) and ADVIA Centaur Vitamin D Total Assay). Samples from the Vitamin D External Quality Survey (DEQAS) and the Standard Reference Material SRM 972 were used for analytical quality control. Results: All methods displayed an acceptable performance with DEQAS samples but immunoassays showed a significant bias against certified materials. Compared to UHPLC, differences were significant for both immunoassays in the deficiency interval but the systematic bias was higher for the ADVIA assay throughout the whole range of concentrations. Conclusion: The prevalence of vitamin D deficiency in clinical practice is assay-dependent and physicians should be aware of the uncertainty associated with vitamin D assessment.

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Background Increased consumer´s interest in health has driven the development of foods that offer specific beneficial effects. The list of foods and ingredients includes essential and non-essential nutrients, plant and marine components, whole foods, microorganisms, microalgae and technological approaches. Traditionally, health outcomes focussed on the prevention of chronic diseases but health targets have expanded to cover areas such as brain health, inflammation, eye health, women´s health, healthy ageing and beauty. Objective This review highlights, from a nutritional biochemistry perspective, differential aspects on designing and interpreting human studies to support the health effects of functional foods. Results Despite the available evidence from in vitro, animal and observational studies, welldesigned human studies are necessary to support the health effects of functional foods. Intervention trials with foods are complex as they imply limitations due to methodological, food-related and host-related factors. The use of responsive, validated and clinically relevant markers becomes essential even though there is a lack of reliable biomarkers of exposure for many bioactives. Furthermore, the effect of modulating factors such as subclinical inflammation, gut microbiota and genetic variability should be taken into account. Multiple indicators may provide a more reliable alternative to assess physiological processes while emerging biomarkers (microRNAs, epigenetic changes) constitute a promising approach. Additionally, the magnitude of the change is critical to support any health effect although interventions may have a limited clinical impact but be epidemiologically relevant. Also, based on the available data, the premise that bioactivescontaining foods are safe may be questionable. Conclusion An integrated approach including multiple biomarkers, genetic variability, effect of gut microbiota and risk/benefit assessment should be used to support the potential health effects of functional foods.

Recent interest in vitamin D has led to a substantial increase in the use of vitamin D supplements. Vitamin D intoxication may be a concern as hypervitaminosis D can result in irreversible calcification of soft tissues so that it is important to detect early markers of vitamin D intoxication. Our aim was to assess the simultaneous presence of biochemical markers of vitamin D toxicity (i.e. hypervitaminosis D, hypercalcemia) and determine the concentrations of 25-OH-vitamin D at which the risk of hypercalcemia, and thus toxicity, might begin.We evaluated retrospectively a 6-year period during which 25.567 samples were assessed for 25-OH-vitamin D status by UHPLC. Hypervitaminosis D was defined at serum 25-OH-vitamin D160 nmol/L. Serum and urine calcium, phosphorus and iPTH were also recorded, if available. Medical history revision was performed in subjects displaying simultaneously hypervitaminosis D and hypercalcemia.Overall, hypervitaminosis D was found in 475 samples (1.86%) of which 51 displayed hypercalcemia (11.1%). A total of 382 samples were identified as the first record of hypervitaminosis D and 39 presented hypercalcemia (10.2%), most of them at 25-OH-vitamin D levels between 161 and 375 nmol/L. Only in 15 subjects, hypercalcemia could be directly attributed to vitamin D and serum 25-OH-vitamin D ranged between 164 and 1139 nmol/l. In no case, serum calcium achieved concentrations considered as critical values (13 mg/dl).Hypercalcemia due to vitamin D represented4% of the total hypervitaminosis D detected and0.1% of the tests performed. However, a highly variable response was observed and most subjects presented hypercalcemia at serum concentrations of 25-OH-vitamin D 375 nmol/L.

Background:Vitamin D deficiency is an important concern in clinical settings although there is no consensus on who should undergo 25-OH-vitamin D testing. We studied the prevalence of vitamin D deficiency before and after introducing adequacy (clinical and biochemical) criteria for testing.Methods:A total of 32,363 tests for 25-OH-vitamin D were retrospectively evaluated. Requests were unrestricted until December 2010 and justification criteria were applied from January 2011. During 6 years, 25,656 samples were analyzed (UHPLC) of which 12,315 were considered the first visit. The prevalence of deficiency was assessed for all the samples and according to the year, sex, season, age, origin of the requests, inclusion of adequacy criteria and consecutive visits.Results:A significant proportion of the requests (25%) were unjustified and less than half of the clinically or biochemically-justified tests displayed serum concentrations indicative of deficiency. Application of adequacy criteria resulted in a non-significant increase in the prevalence of deficiency, both at the first visit (36.5 vs. 41.7, p=0.17) and for all the samples analyzed (32.0 vs. 35.5, p=0.14). The percentage of deficiency decreased in consecutive visits although 2/3 and 41% of the patients remained deficient on the second and third visit, respectively. Moreover, at least 1/5 of sufficient patients at the first test became deficient in subsequent evaluations.Conclusions:A significant proportion of the requests was unjustified by clinical or biochemical criteria. Our data also indicate that clinical and biochemical criteria may be necessary (to be present) to justify vitamin D testing but not sufficient (predictive) to indicate the presence of vitamin D deficiency.

[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.

Objective: To assess the comparability of concentrations of retinol, α- and γ-tocopherols and individual carotenoids in serum and (Li-hep) plasma over a wide range of concentrations. Material and methods: One hundred sixty-six pairs of samples (serum and lithium-heparin plasma) were analyzed by a quality-controlled HPLC method. Means and 95% confidence intervals, differences, interchangeability and the degree of agreement (Bland–Altman plot) were calculated. Results: Distribution of all analytes in the two matrices are comparable and interchangeable with minor quantitative adjustments. Within the range of concentrations assessed, the degree of agreement was high, although some differences were observed for minor components with greater analytical imprecision. Conclusions: The results indicate an acceptable degree of agreement using either of the two matrices for the analytes assessed except possibly for minor blood components. For retinol and α-tocopherol, the comparability and interchangeability of results below the cut-off points for inadequacy need further confirmation.

Natural and added forms of vitamin A (all-trans-retinol, retinyl esters and β -carotene) and vitamin E ( α -tocopherol, γ -tocopherol, tocopheryl acetate) were determined in commercially available dairy products that are frequently consumed. Retinyl esters, β -carotene and α -tocopherol are customarily found in natural dairy products, whereas retinyl and α -tocopheryl acetate occur in most vitamin-fortified products. Lutein, zeaxanthin and γ -tocopherol were found in egg- and oil-containing products, respectively. α -Tocopheryl acetate was found in all vitamin-fortified dairy products analysed, whereas retinyl acetate was only present in fluid milks but not in margarines. Minor amounts of retinyl acetate may occasionally be found in some non-fortified milks, suggesting contamination during processing. In commercially available dairy products in Spain, retinyl palmitate and α -tocopherol contents vary substantially, especially in high fat and vitamin-fortified dairy products. Compared with the label claims in vitamin-fortified products, over-fortification predominates, although over- and under-fortification of both vitamins A and E may occur simultaneously in fluid milks. The wide variation observed in commercially available natural and vitamin-fortified dairy products may be an important source of bias and imprecision in nutritional studies.

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.

Objective We evaluated the effect of intensive insulin therapy and glycemic control in patients with type 1 diabetes on biochemical markers of vitamin A and E. Methods Fifty-seven patients with type 1 diabetes were enrolled in a follow-up study for 3 to 33 mo. At entrance, all patients were on conventional insulin therapy or recently had been diagnosed with the disease. Intensive insulin therapy (multiple daily glycemia records and at least three insulin doses daily) was established, and every 3 to 6 mo patients were screened for clinical, biochemical, and hematologic indexes. Biochemical markers of vitamin A and E nutrition status were measured at each visit by a quality-controlled high-performance liquid chromatography. Results At entrance, serum retinol concentrations, but not the ratio of α-tocopherol to cholesterol, showed a negative correlation with increasing values of HbA1c and insulin dose, neither of which was significant in multiple regression models. With intensive insulin therapy, a trend to normalize parameters of glycemic control (HbA1c and fructosamine) was observed within subjects and on a group level. However, no significant changes were observed in serum retinol or α-tocopherol:cholesterol ratio according to the metabolic control of the disease. Conclusions Patients with type 1 diabetes under intensive insulin therapy tend to normalize the clinical parameters of glycemic control, although this improvement does not significantly affect biochemical markers of vitamin A and E status.

Bioavailability is a critical feature in the assessment of the role of micronutrients in human health. Poorly bioavailable micronutrients like carotenoids may reach significant concentrations in the gastrointestinal tract where they may exert biological actions. We evaluated a simple collection protocol to determine vitamin A, E and carotenoids in microsamples of human faeces as a non-invasive approach for nutritional studies. Microsamples of human faeces were collected using a commercially available device, extracted and analysed on two LC systems. Suitability of the protocol was assessed by evaluating several factors including the effect of simulated colonic conditions and two nutritional scenarios with different dietary components, chemical forms, nutritional goals and target groups. The protocol was reproducible and representative of a faeces sample. The major dietary and serum carotenoids, and several “unidentified” compounds (possibly metabolites) could be detected, and cis-/trans-β-carotene profile reflected dietary intervention. In faeces of neonates, free retinol, retinyl and α-tocopheryl acetate (from infant formula), long-chain fatty acid retinyl esters (from human milk), free γ-tocopherol and α-tocopherol could be detected. Our results show that the analysis of vitamin A, E and carotenoids in microsamples of human faeces is a suitable, non-invasive approach that may provide relevant information regarding responsiveness, nutrient stability and metabolism and may help assess adequacy of chemical forms and delivery systems reaching the colon.

Background Lutein, a non-provitamin A carotenoid, is frequently consumed in the human diet. It is distributed preferentially in certain human tissues (i. e., retina) and shows a high antioxidant activity. Type 1 diabetic patients have been considered to be at risk of increased oxidative stress that may contribute to accelerated atherogenesis and to the microangiopathic complications of the disease. Aim of the study To assess the influence of type 1 diabetes mellitus on the serum depletion rate and bioavailability of lutein. Subjects and Methods Ten type 1 diabetics and eight controls consumed a low carotenoid diet for 21 days and the bioavailability study was performed in 7 diabetics and 5 controls on day 15 with the administration of a capsule of lutein esters from marigold extract. Samples were collected at baseline and on days 1, 2, 3, 6, 11, 15 (eight times during 9 h), 16, 17 and 21. Lutein and other carotenoids were determined by HPLC in triglyceride-rich lipoprotein (TRL) fractions and plasma or serum. Results Serum depletion curve, area of concentrations under time curve (AUC) and final concentration percentages were similar in diabetics and controls. In the bioavailability study, all-trans-lutein increased in both groups and AUC, maxima concentrations in TRL and serum and time required for maxima concentration in serum were similar in diabetics and controls. Conclusions These data suggest that in the group of patients assessed, type 1 diabetes mellitus does not apparently influence the absorption and depletion rate of lutein in serum.

Increased levels of oxidative stress have been implicated in tissue damage and the development of chronic diseases, and dietary antioxidants may reduce the risk of oxidative tissue damage. As part of a European multicentre project, several studies were undertaken with the aim of testing whether the consumption of foods rich in carotenoids reduces oxidative damage to human tissue components. We describe here the serum response of carotenoids and tocopherols upon supplementation with carotenoids from natural extracts (alpha-carotene+beta-carotene, lutein or lycopene; 15 mg/day) and/or with alpha-tocopherol (100 mg/day) in a multicentre, placebo-controlled intervention study in 400 healthy male and female volunteers, aged 25-45 years, from five European regions (France, Northern Ireland, Republic of Ireland, The Netherlands and Spain). Supplementation with alpha-tocopherol increased serum alpha-tocopherol levels, while producing a marked decrease in serum gamma-tocopherol. Supplementation with alpha- + beta-carotene (carotene-rich palm oil) resulted in 14-fold and 5-fold increases respectively in serum levels of these carotenoids. Supplementation with lutein (from marigold extracts) elevated serum lutein (approx. 5-fold), zeaxanthin (approx. doubled) and ketocarotenoids (although these were not present in the supplement), whereas lycopene supplementation (from tomato paste) resulted in a 2-fold increase in serum lycopene. The isomer distributions of beta-carotene and lycopene in serum remained constant regardless of the isomer composition in the capsules. In Spanish volunteers, additional data showed that the serum response to carotenoid supplementation reached a plateau after 4 weeks, and no significant side effects (except carotenodermia) or changes in biochemical or haematological indices were observed throughout the study. This part of the study describes dose-time responses, isomer distribution, subject variability and side effects during supplementation with the major dietary carotenoids in healthy subjects.

OBJECTIVE: Epidemiologic studies have suggested that dietary intake and plasma concentrations of antioxidants have an inverse relation with coronary heart disease. To test whether fat-soluble antioxidants can play a role against the occurrence of myocardial infarction (MI), we measured plasma levels of retinol, tocopherols, and individual carotenoids in MI patients. METHODS: A case-control and follow-up study of patients in the Mostoles area (Madrid, Spain). One hundred six patients (62 after 1 y) and 104 control subjects participated in the study. Blood samples were collected after overnight fast or during the first 24 h of MI onset for biochemical profiles of retinol, α- and γ-tocopherols, and carotenoid by means of a quality-controlled high-performance liquid chromatography. RESULTS: During the acute phase after MI onset, plasma levels of retinol, γ-tocopherol, and xanthophylls (lutein/zeaxanthin and β-cryptoxanthin) decreased, whereas α-tocopherol, α-carotene, β-carotene, and lycopene showed levels similar to those of control subjects. Logistic regression analysis showed low concentrations of γ-tocopherol (and retinol) in plasma as the only statistically significant factor associated with MI, after adjusting for traditional risk factors. However, 1 y later, the MI patients showed a general improvement in plasma lipids and fat-soluble antioxidant status, and none of the analytes was associated with MI. CONCLUSIONS: The decreased plasma status of retinol, γ-tocopherol, and xanthophylls during the acute phase of MI normalized the year after the MI event, suggesting that most subjects had followed an overall healthier lifestyle and dietary pattern. The results also raise concerns on the usefulness of these plasma compounds as specific, relevant, and predictive markers in relation to coronary heart disease.

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).

Carotenoids are mainly present in vegetables and fruits and display several important biological actions (provitamin A, antioxidant and anticarcinogenic activities and immunomodulation). To assess the intake of carotenoids and their potential health benefits in humans, it is necessary to have reliable information about carotenoid content in food. Saponification is commonly used during analysis of carotenoids in vegetables and fruits to release esterified xanthophylls and to eliminate interfering substances. It is a time-consuming step and involves the use of considerable volumes of solvents in the partition. To reduce the time and costs of the saponification process, we compare the reliability of a “shortcut” (small volumes, vortex 3 min, 20% potassium hydroxide, hexane/methylene chloride extraction) with that of a “standard” protocol (stirring 30 min, 5% potassium hydroxide, petroleum ether/diethyl ether extraction twice plus washings). Accuracy and precision were assessed according to studies of certified reference materials for carotenoid analysis in foods. For lutein, zeaxanthin, α -cryptoxanthin, β -cryptoxanthin, lycopene, α -carotene andβ -carotene, the “shortcut” showed within-day and between-day variability similar to or lower than that of the “standard” protocol, comparable deviation from the assigned values and higher recovery for carotenes, especially for lycopene. Advantages of the “shortcut” include considerably reduced time, higher overall recoveries and lower solvent expenditure, resulting in a reduction in time and total cost of up to 80–90% without loss of accuracy or precision.

Type 1 diabetics are considered to be at risk for marginal status of vitamin A, although they frequently present higher serum levels of provitamin A carotenoids, suggesting an impaired bioconversion into retinol. Our aim was to assess the bioavailability of provitamin A carotenoids (absorption and conversion into retinol) in these subjects compared to non-diabetics using the post-prandial triacylglycerol-rich lipoprotein fractions (TRL) model and the depletion rate in serum after 21 days on a low-carotenoid diet. The analysis was performed by a quality-controlled HPLC method. In both groups, total retinyl esters accounted for approximately 60% of the total amount absorbed from the capsule, retinyl palmitate being the predominant ester and efficacy of conversion (AUC total esters/AUC α-+β-carotene) was similar. Markers of glycemic control were correlated with absorption but not with conversion into retinol. Serum depletion rate in diabetics was not significantly different from control subjects. Provitamin A carotenoids bioavailability in type 1 diabetics is, at least qualitatively, similar to control subjects.

Lutein, a non-provitamin A xanthophyll, is widely distributed in fruits and vegetables frequently consumed. In human serum, lutein is transported by lipoproteins and selectively accumulated in certain tissues (eg the retina). Epidemiological studies suggest that high intake and serum levels of lutein are associated with a lower risk of cataracts and age-related maculopathy. Subjects diagnosed with cataracts (CA; n =5) or age-related macular degeneration (ARMD; n =5) agreed to take three lutein capsules per week. Each capsule potentially provided about 12 mg of all-trans-lutein, 3 mg of 13/15-cis-lutein and 3.3mg of α-tocopherol, as revealed by HPLC. Zeaxanthin was not detected. Average supplementation time was 13 months (range 4-20 months) for ARMD subjects and 26 months (range 16-36 months) for CA subjects. Blood samples for carotenoid analysis were collected every 3 months, coinciding with ophthalmological revision. In serum, concentrations of lutein, 13-cis-lutein and two ketocarotenoids increased significantly. Maximum increments were observed at 3-6 months, reaching levels above the 95th percentile of the reference population (>0.44 μmol l -1 ). Ophthalmological evaluation showed an average increment in visual acuity of 0.4, and glare sensitivity also improved. No significant side effects such as hypercarotenemia, carotenodermia or changes in biochemical or haematological profile were observed. Thus, lutein supplementation at achievable dietary levels increased and maintained serum lutein levels, which were associated with an improvement in the visual function of the patients. Des etudes epidemiologiques suggerent que la supplementation en luteine et des taux seriques eleves sont associes a des risques de cataractes et de degenerescences maculaires plus faibles. Des sujets atteints ont ete supplementes par voie orale, a raison de 3 prises par semaine. Chaque prise contient 12mg de luteine all-trans, 3 mg de 13/15-cis- luteine, et 3,3 mg d'α-tocopherol, pas de zeaxanthine. La duree du traitement etant de 13 mois en moyenne pour la cataracte et de 26 mois pour la degenerescence maculaire. Le dosage des carotenoides sanguins a ete effectue tous les 3 mois en meme temps que les controles ophtamologiques.Aucun effet secondaire n'a ete note, et les fonctions visuelles des patients sont ameliorees.

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.

1. Patients with insulin-dependent diabetes mellitus are classified among the groups at risk for low vitamin status, and recent studies suggest that some degree of supplementation with antioxidants may be beneficial in helping to prevent certain long-term complications of diabetes mellitus. Our objective was to compare the status of the fat-soluble vitamins and antioxidant-related compounds in patients with well-defined insulin-dependent diabetes mellitus with that of their first-degree relatives, controlling seasonal and analytical variability as factors influencing the interpretation of the data. 2. Fifty-four patients with insulin-dependent diabetes mellitus, 214 non-diabetic, first-degree relatives (controls) and 236 unrelated controls were analysed for retinol, tocopherols (α and γ) and main carotenoids in serum (β-carotene, α-carotene, β-cryptoxanthin, lutein, zeaxanthin and lycopene) by means of a validated HPLC method. 3. Insulin-dependent diabetes mellitus was associated with lower retinol levels and higher levels of β-carotene, α-carotene and β-cryptoxanthin than sex-matched, first-degree relatives. α-Tocopherol, the α-tocopherol/cholesterol ratio, γ-tocopherol, lutein, zeaxanthin and lycopene showed no differences. Retinol and β-carotene were the variables most closely associated with diabetes. 4. Patients with insulin-dependent diabetes mellitus showed lower serum retinol status together with higher concentrations of provitamin-A carotenoids. Serum fat-soluble antioxidant levels were greater than or equal to those in controls. According to the serum status observed, individuals with diabetes do not require supplementation with α-tocopherol or carotenoids, although the need for retinol supplementation in patients with marginal serum levels should be evaluated.

Background and aim Post-menopausal women are at higher risk of cardiovascular disease and bone demineralization. Phytosterols (PS) may be used for hypercholesterolemia in some groups and β-cryptoxanthin (β-Cx) displays a unique anabolic effect on bone. Our aim was to assess the changes in cardiovascular and bone turnover markers from the oral intake of β-Cx and PS in post-menopausal women. Methods and results A randomized, double-blind, crossover study with β-Cx (0.75 mg/day) and PS (1.5 g/day), single and combined, was performed in 38 postmenopausal women. Diet was supplemented with 1 × 250 mL milk-based fruit drink/day for 4 weeks with a wash-out period of 4-weeks in between. Serum β-Cx and PS were determined by UPLC and CG-FID respectively. Outcome variables included markers of bone turnover and cardiovascular risk. Biological effect was assessed by paired t test and generalized estimating equations analysis that included the previous treatment, the order of intervention and the interactions. The intake of beverages containing β-Cx and PS brought about a significant increase in serum levels of β-Cx, β-sitosterol and campesterol. Intervention caused changes in almost all the markers while the order, previous treatment and the interaction did not reach statistical significance. Only the intake of the beverage containing β-Cx plus PS brought about significant decreases in total cholesterol, c-HDL, c-LDL and bone turnover markers. Conclusions β-Cx improves the cholesterol-lowering effect of PS when supplied simultaneously and this combination may also be beneficial in reducing risk of osteoporosis. Clinical Trial Registry ClinicalTrials.gov number NCT01074723.

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.

Diet constitutes a key modifiable factor in the development and maintenance of health. Malnutrition (both over- and under-nutrition) plays a critical role in morbidity and mortality, and therefore the evaluation of nutritional status is a key point to improve the health of the individuals and popula...

Obesity constitutes a growing health problem, and surgical treatment of severe obesity is increasingly used. Nutrient deficiencies are common following bariatric surgery and the evidence indicates a progressive increase in the incidence and severity of the deficiency of certain vitamins and related clinical conditions. Because of the potential role of carotenoids in disease prevention, our aim was to assess the carotenoid status in candidates for obesity surgery and the time-course changes following two bariatric procedures.Seventy-five candidates for bariatric surgery (17 men, 58 women; age 43 ± 10 years) and a total of 362 serum samples after obesity surgery (i.e., Roux-en-Y gastric bypass (n = 187) and biliopancreatic diversion (n = 175)) were consecutively collected and assessed. Retinol, α- and γ-tocopherol, 25-OH-vitamin D3, lutein, zeaxanthin, α- and β-cryptoxanthin, lycopene (trans and cis), α- and β-carotene (trans and cis) were analyzed by high-performance liquid chromatography.Mean serum levels of carotenoids in candidates for obesity surgery were within the reference values reported in controls and seasonal variations were present in several analytes. After surgery, and regardless of the type of intervention, all serum carotenoids dropped following first-order kinetics. Cis/trans ratio of lycopene and β-carotene did not change after surgery, over the time or between surgical procedures. On a long-term, serum carotenoids were at or below fifth percentile of reference groups.The chronic low levels of carotenoids in these patients compromise their availability to tissues, constituting an additional risk factor for other clinical conditions. Dietary advice on carotenoid-rich, fortified foods or supplements should be also evaluated in these patients.

The available evidence indicates a progressive increase in the incidence and severity of the deficiency of certain vitamins and related clinical conditions (i.e., metabolic bone disease). Because of the potential role of fat-soluble vitamins and carotenoids in bone metabolism, our aim was to assess the time-course changes of fat-soluble vitamins and serum markers of bone metabolism in candidates for obesity surgery and following two bariatric procedures.Sixty-five candidates for bariatric surgery and 150 serum samples after obesity surgery (i.e., Roux-en-Y gastric bypass, n = 85; biliopancreatic diversion, n = 65) were consecutively analyzed over a period of more than 2 years. Retinol, α- and γ-tocopherol, 25-OH-vitamin D3, β-cryptoxanthin, and β-carotene were analyzed by high-performance liquid chromatography. Calcium, phosphorus, alkaline phosphatase, intact parathyroid hormone (iPTH), osteocalcin, beta-crosslaps, and N-terminal peptide of procollagen I (P1NP) were determined using commercial kits.Retinol, β-cryptoxanthin, β-carotene, and α- and γ-tocopherol levels were significantly lower in post-surgery samples while osteocalcin, b-crosslaps, and P1NP were significantly increased. Along the time and regardless of the surgical procedure, P1NP, b-crosslaps, and osteocalcin increased during the first 12-24 months but declined afterward. 25-OH-vitamin D increased during the first 12 months and tended to decrease afterward while iPTH remained constant or decreased but increased after 1 year in both groups. Vitamin A remained constant but α- and γ-tocopherol, β-cryptoxanthin, and β-carotene decreased in both groups.In addition to the nutritional assessment, regular monitoring of bone markers seems necessary in these patients and the early introduction of preventive strategies (i.e., the use of antiresorptive agents) should be evaluated.

Objective To compare two methods for assessing vitamin D status in clinical settings. Design and methods 25-OH-vitamin D status was measured in 120 patients by HPLC and a commercial immunoassay. Results Only 53% of the subjects considered as vitamin D sufficient by immunoassay fell within this category according to HPLC although subjects with concentrations above 75 nmol/L, regardless of the method used, presented normal PTH concentrations. Conclusions Both methods are not exchangeable to classify subjects based on unique cut-offs but they are comparable when interpreted in relation with PTH concentrations.

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, 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

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.

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).

As part of a European project to test if 'the consumption of diets rich in carotenoids reduces oxidative damage to human tissue components', 15 mg/day lutein and 100 mg/day alpha-tocopherol were supplied to ten volunteers. alpha-Tocopherol (alone and combined) provoked an increase in alpha-tocopherol and a decrease in gamma-tocopherol serum levels. Lutein levels increased during the first month (given alone) in all controls but they behaved very differently during the following 3 months. Ketocarotenoids (not supplied) increased whereas anhydroluteins (not supplied) did not change, suggesting that they are formed in vivo.