A decrease in iron status in young healthy women after long-term daily consumption of the recommended intake of fibre-rich wheat bread.

BACKGROUND: Fibrerich bread and cereals are included in the recommendations of a healthy diet. OBJECTIVE: To measure the effects of long-term consumption of the recommended intake of fibre-rich wheat bread on the iron status of young healthy women with adequate iron stores. DESIGN: Four-months intervention study including healthy female subjects assigned into two groups provided daily with 300 g of fibre-rich wheat bread, prepared with or without phytase as a supplement to their habitual diet. SUBJECTS: Forty-one women aged 24.8 +/- 3.8 years (mean +/- SD) and an average BMI of 22.0 +/- 2.9 kg/m2 participated. Baseline values for serum ferritin were 45 microg/L, 22-83 (geometric mean, range) and for haemoglobin 132 g/L, 119-148 (arithmetic mean, range), respectively. RESULTS: Distribution of energy intake from protein, fat and carbohydrate, and daily intake of dietary fibre and iron were similar in the two groups and within the recommended levels. There was no effect of the phytase added to the wheat bread on the iron status of the subjects, but an effect of the intervention period. Serum ferritin and haemoglobin levels were significantly reduced by 12 +/- 1.1 microg/L (27%) (P < 0.001) and 2 +/- 0.8 g/l (1.5%) (mean +/- SE) (P < 0.05) respectively, after four months of intervention. CONCLUSIONS: The present long-term study indicates that consumption of the recommended daily intake of fibre-rich wheat bread results in an impairment of iron status in women with initially sufficient iron stores. Reduction of the phytic acid concentration in the bread was not sufficient to maintain iron status.

Pork meat: a good source of selenium?

The knowledge about Se bioavailability from animal food is sparse. This study was therefore initiated in order to evaluate the bioavailability of Se from pork meat in humans. Twelve male volunteers (age 21-30 years) participated in a randomised crossover study with strictly controlled diet containing 170 g pig meat/10 MJ per day and 106 +/- 13 microg Se/d for 3 x 3 weeks. Complete faecal and urinary collections were made during the last week of each period. Bioavailability was evaluated from absorption and retention of Se and changes in plasma Se concentration and blood glutathione peroxidase (GSHPx) activity. The apparent absorption of Se was 94 +/- 2% (100 +/- 13 microg/d). Faecal and urinary excretion were 7 +/- 1 microg/d and 39 +/- 21 microg/d, respectively, resulting in a retention of 61 +/- 24 microg/d. The diet intervention did not affect plasma Se concentration and GSHPx activity. Absorption and retention of Se from pig meat were high suggesting a high availability. However, the availability of pig meat Se for blood Se protein appears to be low.

The 6-a-day study: effects of fruit and vegetables on markers of oxidative stress and antioxidative defense in healthy nonsmokers.

BACKGROUND: Fruit and vegetables contain both nutritive and nonnutritive factors that might contribute to redox (antioxidant and prooxidant) actions. OBJECTIVE: We investigated the relative influence of nutritive and nonnutritive factors in fruit and vegetables on oxidative damage and enzymatic defense. DESIGN: A 25-d intervention study with complete control of dietary intake was performed in 43 healthy male and female nonsmokers who were randomly assigned to 1 of 3 groups. In addition to a basic diet devoid of fruit and vegetables, the fruit and vegetables (Fruveg) group received 600 g fruit and vegetables/d; the placebo group received a placebo pill, and the supplement group received a vitamin pill designed to contain vitamins and minerals corresponding to those in 600 g fruit and vegetables. Biomarkers of oxidative damage to protein and lipids and of antioxidant nutrients and defense enzymes were determined before and during intervention. RESULTS: Plasma lipid oxidation lag times increased during intervention in the Fruveg and supplement groups, and the increase was significantly higher in the former. Plasma protein carbonyl formation at lysine residues also increased in both of these groups. Glutathione peroxidase activity increased in the Fruveg group only. Other markers of oxidative damage, oxidative capacity, or antioxidant defense were largely unaffected by the intervention. CONCLUSIONS: Fruit and vegetables increase erythrocyte glutathione peroxidase activity and resistance of plasma lipoproteins to oxidation more efficiently than do the vitamins and minerals that fruit and vegetables are known to contain. Plasma protein carbonyl formation at lysine residues increases because of the vitamins and minerals in fruit and vegetables.

A solid dietary fat containing fish oil redistributes lipoprotein subclasses without increasing oxidative stress in men.

There is a demand and need for healthy solid dietary fats. However, synthetic fats can be tailored to contain specific physiologic properties. Our goal was to design dietary solid test fats that would be both beneficial to the atherogenic lipid profile and stable against lipid peroxidation. Sixteen men (age 35-75 y) substituted 80 g of their normal dietary fat intake with test fat for two periods of 21 d each in a double-blind, randomized, crossover study. Although solid, both test fats were low in cholesterol-raising SFA. Test fat "F" contained 5 g/100 g long chain (n-3) fatty acids matched by oleic acid in test fat "O." Plasma total triacylglycerol (TAG), VLDL TAG, cholesterol in VLDL, and intermediate density lipoproteins (IDL) were lower (P < 0.05), whereas apolipoprotein (apo) B of the large LDL-2 (d = 1031-1042 g/L) subclass, and cholesterol of HDL(2b) subclass, were higher after intake of F than O fat (P < 0.05). There was no difference in the effect on in vivo oxidation measured as the ratio of plasma isoprostanes F(2) to arachidonic acid and urinary isoprostanes, whereas the vitamin E activity/plasma total lipids ratio was higher after intake of F than O (P = 0.008). In conclusion, a solid dietary fat containing (n-3) PUFA decreased plasma TAG, VLDL, and IDL cholesterol, and redistributed lipoprotein subclasses in LDL and HDL, with a higher concentration of the larger and less atherogenic subfractions. These changes took place without an increase in oxidative stress as measured by in vivo markers.

Does fat in milk, butter and cheese affect blood lipids and cholesterol differently?

OBJECTIVE: To compare the effects of isoenergetic amounts of milk, cheese and butter (adjusted to the same content of lactose and casein) on fasting and postprandial blood lipids and lipoproteins, and on postprandial glucose and insulin response. DESIGN: The experiments were designed to provide 20% of total energy from dairy fat, as either whole milk, mean (+/-SD) 2164 (+/-97) g, butter 93 (+/-4) g, and hard cheese 305 (+/-45) g, which were served to 14 healthy young men for three periods of three weeks each, separated by washout periods, in a randomized, cross-over study with strictly controlled dietary intake. Fasting blood samples were taken at the end of the study periods. Measurements of the postprandial effect of the three different dairy test products (0.7 g of milk fat/kg body weight) were carried out on day 4 of each intervention period. Blood samples were taken before and at 2, 4, 6 and 8 hours following intake of the meals. RESULTS: Fasting LDL cholesterol concentration was significantly higher after butter than cheese diet (p = 0.037), with a borderline significant difference in total cholesterol (p = 0.054) after the experimental periods of three weeks. Postprandial glucose showed a higher response after cheese diet than after milk diet (p = 0.010, diet x time interaction). CONCLUSIONS: A different effect of fat in milk and butter could not be confirmed in this study. The moderately lower LDL cholesterol after cheese diet compared to butter diet should be investigated further.

Effects of medium-chain fatty acids and oleic acid on blood lipids, lipoproteins, glucose, insulin, and lipid transfer protein activities.

BACKGROUND: Dietary medium-chain fatty acids (MCFAs) are of nutritional interest because they are more easily absorbed from dietary medium-chain triacylglycerols (MCTs) than are long-chain fatty acids from, for example, vegetable oils. It has generally been claimed that MCFAs do not increase plasma cholesterol, although this claim is poorly documented. OBJECTIVE: We compared the effects of a diet rich in either MCFAs or oleic acid on fasting blood lipids, lipoproteins, glucose, insulin, and lipid transfer protein activities in healthy men. DESIGN: In a study with a double-blind, randomized, crossover design, 17 healthy young men replaced part of their habitual dietary fat intake with 70 g MCTs (66% 8:0 and 34% 10:0) or high-oleic sunflower oil (89.4% 18:1). Each intervention period lasted 21 d, and the 2 periods were separated by a washout period of 2 wk. Blood samples were taken before and after the intervention periods. RESULTS: Compared with the intake of high-oleic sunflower oil, MCT intake resulted in 11% higher plasma total cholesterol (P = 0.0005), 12% higher LDL cholesterol (P = 0.0001), 32% higher VLDL cholesterol (P = 0.080), a 12% higher ratio of LDL to HDL cholesterol (P = 0.002), 22% higher plasma total triacylglycerol (P = 0.0361), and higher plasma glucose (P = 0.033). Plasma HDL-cholesterol and insulin concentrations and activities of cholesterol ester transfer protein and phospholipid transfer protein did not differ significantly between the diets. CONCLUSIONS: Compared with fat high in oleic acid, MCT fat unfavorably affected lipid profiles in healthy young men by increasing plasma LDL cholesterol and triacylglycerol. No changes in the activities of phospholipid transfer protein and cholesterol ester transfer protein were evident.

No effect of 600 grams fruit and vegetables per day on oxidative DNA damage and repair in healthy nonsmokers.

In several epidemiological studies, high intakes of fruits and vegetables have been associated with a lower incidence of cancer. Theoretically, intake of antioxidants by consumption of fruits and vegetables should protect against reactive oxygen species and decrease the formation of oxidative DNA damage. We set up a parallel 24-day dietary placebo-controlled intervention study in which 43 subjects were randomized into three groups receiving an antioxidant-free basal diet and 600 g of fruits and vegetables, or a supplement containing the corresponding amounts of vitamins and minerals, or placebo. Blood and urine samples were collected before, once a week, and 4 weeks after the intervention period. The level of strand breaks, endonuclease III sites, formamidopyrimidine sites, and sensitivity to hydrogen peroxide was assessed in mononuclear blood cells by the comet assay. Excretion of 7-hydro-8-oxo-2'-deoxyguanine was measured in urine. The expressions of oxoguanine glycosylase 1 and excision repair cross complementing 1 DNA repair genes, determined by real-time reverse transcription-PCR of mRNAs, were investigated in leukocytes. Consumption of fruits and vegetables or vitamins and minerals had no effect on oxidative DNA damage measured in mononuclear cell DNA or urine. Hydrogen peroxide sensitivity, detected by the comet assay, did not differ between the groups. Expression of excision repair cross complementing 1 and oxoguanine glycosylase 1 in leukocytes was not related to the diet consumed. Our results show that after 24 days of complete depletion of fruits and vegetables, or daily ingestion of 600 g of fruit and vegetables, or the corresponding amount of vitamins and minerals, the level of oxidative DNA damage was unchanged. This suggests that the inherent antioxidant defense mechanisms are sufficient to protect circulating mononuclear blood cells from reactive oxygen species.

Effect of diets based on foods from conventional versus organic production on intake and excretion of flavonoids and markers of antioxidative defense in humans.

Different food production methods may result in differences in the content of secondary metabolites such as polyphenolic compounds. The present study compared conventionally (CPD) and organically produced (OPD) diets in a human crossover intervention study (n = 16) with respect to the intake and excretion of five selected flavonoids and effect on markers of oxidative defense. The urinary excretion of quercetin and kaempferol was higher after 22 days of intake of the OPD when compared to the CPD (P < 0.05). The excretions of flavonoids in urine as a percentage of intake (0.6-4%) were similar after both interventions. Most markers of antioxidative defense did not differ between the diets, but intake of OPD resulted in an increased protein oxidation and a decreased total plasma antioxidant capacity compared to baseline (P < 0.05). Some varietal difference was seen in the study, and because selection of more resistant varieties is of central importance to organic farming, it cannot be excluded that the observed effects originate from these differences. The food production method affected the content of the major flavonoid, quercetin, in foods and also affected urinary flavonoids and markers of oxidation in humans.

Effect of individual dietary fatty acids on postprandial activation of blood coagulation factor VII and fibrinolysis in healthy young men.

BACKGROUND: Hypertriglyceridemia may represent a procoagulant state involving disturbances to the hemostatic system. Plasminogen activator inhibitor type 1 (PAI-1) is increased in the presence of hypertriglyceridemia. Free fatty acids (FFAs) in plasma may promote factor VII (FVII) activation. OBJECTIVE: We tested the hypothesis that FVII activation would be less after consumption of saturated fatty acids than after other fatty acids. DESIGN: The effects of 6 matching dietary test fats, rich in stearic (S), palmitic (P), palmitic + myristic (M), oleic (O), trans 18:1 (T), and linoleic (L) acid, respectively, on the postprandial lipid and hemostatic profile (after 2, 4, 6, and 8 h) were investigated in 16 young men. High-fat meals (1 g fat/kg body wt; 43% from the test fatty acid) were served in the morning on 6 separate days. RESULTS: All fats increased FVII activation. The S fat resulted in a lower increase in activated FVII (FVIIa) than did the T fat and in a lower FVII coagulant activity (FVII:c) than did the O fat (P < 0.02, diet x time interaction). When the data were pooled, the saturated (S, P, and M) test fats resulted in a smaller postprandial increase in FVIIa (P = 0.036, diet effect), a smaller increase in FVII:c (P < 0.001, diet x time interaction), a greater rise in tissue plasminogen activator concentrations (P = 0.028, diet effect), and a tendency to a greater postprandial decline in PAI-1 (P = 0.06, diet effect) compared with the unsaturated test fats (O, T, and L). The increase in FVIIa was not significantly associated with the level of lipemia, plasma FFAs, or plasma lipoprotein lipase activity. CONCLUSION: Our results indicate a lesser increase in FVIIa after the consumption of saturated fats, especially the S fat, than after unsaturated test fats.

Simultaneous determination of hydroxycinnamates and catechins in human urine samples by column switching liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry.

A quantitative liquid chromatography mass spectrometry (LC-MS) methodology with online sample clean up by column switching is described for the simultaneous determination of the hydroxycinnamates, caffeic acid and chlorogenic acid, and of the catechins, epicatechin and catechin in human urine samples. Enzymatically treated urine samples were directly injected onto the LC-MS system, where sample clean up was performed by a reversed-phase Zorbax 300SB C(3) column and selective elution of the target compounds onto a Zorbax SB C(18) column resulted in final separation prior to detection by atmospheric pressure chemical ionization (APCI) MS using single ion monitoring (SIM) in negative mode. Linear calibration graphs were achieved in the dynamic range of 10-1000 ng/ml urine. The inter- and intraassay coefficients of variation (C.V.%) for the analysis of the four compounds in quality control urine samples were between 7.8 and 10.9, n=17 (reproducibility), and the repeatability of the assay was between 2.5 and 5.0% (n=12). Analyses of urine samples from a human dietary intervention study with intake of 600 g of fruits and vegetables were demonstrated. To our knowledge, this is the first method described that allows simultaneous determination of both hydroxycinnamates and catechins in biological samples.

A novel dual radio- and stable-isotope method for measuring calcium absorption in humans: comparison with the whole-body radioisotope retention method.

BACKGROUND: Dietary calcium absorption can be determined only with the use of isotope techniques. Currently used isotope techniques require exclusive equipment or are not true tracer approaches. OBJECTIVE: The objective was to compare a dual-isotope method combining radioisotopes and stable isotopes with a whole-body radioisotope retention method for measuring calcium absorption. DESIGN: Seven healthy adults aged 21-27 y consumed a test meal containing 63 +/- 14 (macro x +/- SD) mg Ca together with a water solution of (47)Ca (0.11 MBq). One hour after ingestion, 18 mg (44)Ca was administered intravenously. All feces and urine were collected for 5 and 6 d, respectively. Calcium absorption was estimated from whole-body retention of the radioisotope 12 times over 3 wk after ingestion and from the excretion of (47)Ca and (44)Ca in a 24-h urine sample collected on day 2. (44)Ca in urine was determined by inductively coupled plasma mass spectrometry. RESULTS: Mean (+/- SD) calcium absorption was 75 +/- 9% with the dual-isotope method and was 74 +/- 8% with the whole-body radioisotope retention method. There was a high degree of agreement between the methods. CONCLUSION: The dual-isotope method is a valid approach for measuring calcium absorption from a single meal.

Increasing the cooking temperature of meat does not affect nonheme iron absorption from a phytate-rich meal in women.

The effect of increasing cooking temperatures of meat on nonheme iron absorption from a composite meal was investigated. Cysteine-containing peptides may have a role in the iron absorption enhancing effect of muscle proteins. Heat treatment can change the content of sulfhydryl groups produced from cysteine and thereby affect iron absorption. Twenty-one women (25 +/- 3 y) were served a basic meal without meat and two other meals consisting of the basic meal plus 75 g of pork meat cooked at 70, 95 or 120 degrees C. The meals were extrinsically labeled with (55)Fe or (59)Fe. Iron absorption was determined from measurements of whole-body (59)Fe retention and the activity of (55)Fe and (59)Fe in blood samples. Nonheme iron absorptions were 0.9 (0.5-4.0)% (P = 0.06), 0.7 (0.4-3.9)% (P = 0.1) and 2.0 (1.3-3.1)% (P < 0.001) greater when meat cooked at 70, 95 or 120 degrees C, respectively, was added to the basic meal. Increasing the cooking temperature of meat did not impair nonheme iron absorption compared with cooking at 70 degrees C. Because the cysteine content of meat decreased with increasing cooking temperature, this argues against a specific contribution of sulfhydryl groups from cysteine residues in the promotion of nonheme iron absorption by meat proteins.

Nonheme-iron absorption from a phytate-rich meal is increased by the addition of small amounts of pork meat.

BACKGROUND: Muscle tissue from various sources is known to promote nonheme-iron absorption. However, systematic studies of the dose dependency of this effect of meat on iron absorption from an inhibitory meal with low amounts of meat are lacking. OBJECTIVE: We investigated the dose-response effect of small amounts of meat on nonheme-iron absorption from a meal presumed to have low iron bioavailability. DESIGN: Forty-five healthy women with a mean (+/-SD) age of 24 +/- 3 y were randomly assigned to 1 of 3 groups, each of which was served (A) a basic meal (rice, tomato sauce, pea purée, and a wheat roll) and (B) the basic meal with either 25, 50, or 75 g pork (longissimus muscle). Meal A contained 2.3 mg nonheme iron, 7.4 mg vitamin C, and 220 mg (358 micro mol) phytate. Each meal was served twice, and the order of the meals was ABBA or BAAB. The meals were extrinsically labeled with (55)Fe or (59)Fe. Iron absorption was determined from measurements of (59)Fe whole-body retention and the activity of (55)Fe and (59)Fe in blood samples. RESULTS: Twenty-five grams meat did not increase nonheme-iron absorption significantly (P = 0.13), whereas absorption increased 44% (P < 0.001) and 57% (P < 0.001), respectively, when 50 and 75 g meat were added to the basic meal. In absolute values, this corresponds to an absorption that was 2.6% and 3.4% higher, respectively, than that with the basic meal after adjustment of the data to a level of 40% absorption from a reference dose. CONCLUSION: Small amounts of meat (>or=50 g) significantly increase nonheme-iron absorption from a phytate-rich meal low in vitamin C.

Inter-individual variation, seasonal variation and close correlation of OGG1 and ERCC1 mRNA levels in full blood from healthy volunteers.

The mRNA levels of the nucleotide excision DNA repair gene ERCC1 and the base excision DNA repair gene OGG1 were quantified in 43 healthy volunteers in a dietary intervention trial as markers for the DNA repair capacity. Nine samples were collected from each subject over a period of 52 days. Sampling took place from January to May. The mRNA levels of OGG1 and ERCC1 correlated closely (r = 0.86, P << 0.0001) after normalization to either 18S ribosomal RNA or to beta-actin mRNA. The levels of OGG1 and ERCC1 mRNA were relatively constant within an individual with intra-individual correlation (R(2) = 0.45-0.46) in a General Linear Model. The amounts of ERCC1 and OGG1 relative to 18S RNA were doubled in May compared with January. This coincided with an increase in the monthly influx of sunlight from 18 MJ/m(2) in January to 242 MJ/m(2) in May. The mRNA levels of both ERCC1 and OGG1 were positively correlated to the average daily influx of sunlight in the previous 30 and 5 days (r = 0.49; r = 0.37, respectively, P << 0.001). There were no significant effects of the dietary interventions. The inter-individual variation was 5-10-fold, which is more than the observed 2-3-fold seasonal variation. Thus, despite seasonal variation of the individual mRNA levels, the inter-person variation is still far larger than the intra-person variation, supporting the use as biomarkers.

Different effects of diets rich in olive oil, rapeseed oil and sunflower-seed oil on postprandial lipid and lipoprotein concentrations and on lipoprotein oxidation susceptibility.

Elevated concentrations of fasting and non-fasting triacylglycerol-rich lipoproteins (TRL) as well as oxidative changes of lipoproteins may increase the risk of ischaemic heart disease. To compare the effects of different diets rich in unsaturated fatty acids on the concentrations and in vitro oxidation of fasting and postprandial lipoproteins eighteen males consumed diets enriched with rapeseed oil (RO), olive oil (OO), or sunflower-seed oil (SO) in randomised order for periods of 3 weeks followed by a RO test meal. In the postprandial state the concentrations of cholesterol and triacylglycerol (TAG) in TRL were higher after consumption of OO compared with RO and SO (P<0.04), possibly related to differences in the fasting state. The propagation rates for VLDL and LDL oxidation were higher in the postprandial compared with the fasting state irrespective of diet. In the fasting state, the propagation rates were highest after SO (P<0.001), and in the postprandial state, SO gave rise to a shorter VLDL lag time (P=0.03) and a higher propagation rate than OO consumption (P=0.04). Overall, the SO diet resulted in a higher postprandial propagation rate of LDL (P<0.001) compared with RO and OO, while there was no effect of diet on LDL oxidation lag time. Our results suggest that RO and SO diets lower the postprandial cholesterol and TAG concentrations compared with OO, while RO and OO diets result in similar and lower in vitro susceptibility to oxidation of lipoproteins than SO.