Super-complex mixtures of aliphatic- and aromatic acids may be common degradation products after marine oil spills: A lab-study of microbial oil degradation in a warm, pre-exposed marine environment.

When assessing oil spills in marine environments, focus has often been on describing degradation and removal of hydrocarbons. However, more and more attention is now given to the formation of mineral oil transformation products, and their potential toxicity and persistency in the environment. The aim of this study was to investigate the formation of dissolved acidic degradation products from crude oil in sea water from the Persian Gulf in a lab-experiment. A super-complex mixture of acidic degradation products was formed in the water phase and compound groups of aliphatic acids, monocyclic aromatic acids-, and polycyclic aromatic acids were identified. More specifically, alkylated PAHs were biodegraded to a high number of aromatic, carboxylic acids by hydroxylation of the alkyl side chains. These degradation products are more bioavailable than their parent compounds, and may therefore constitute a new group of contaminants that should be considered in oil spill assessments.

Optimization and validation of a derivatization method with boron trifluoride in ethanol for analysis of aromatic carboxylic acids in water.

Gas-chromatography (GC) analysis of carboxylic acids is limited by the high polarity and low volatility of most of these compounds. Boron trifluoride (BF3) mediated alkylation reactions is one of the most commonly used derivatization methods for making carboxylic acids GC compatible. A semi-automated BF3·EtOH (ethanol) derivatization method was optimized for comprehensive two-dimensional gas chromatography high-resolution mass spectrometry (GC × GC-HR MS) analysis of carboxylic acids in solid phase extraction (SPE) extracts of oil polluted water. The optimal derivatization method were found to be with addition of 300 µL BF3·EtOH per 200 µL sample and reaction at 75 °C for 24 h. Derivatives of eight selected acids (aliphatic, mono- and di-aromatic) were stable over 12 h with relative standard deviations (RSDs) of 2.0-10.7 %, the derivatization method was repeatable (RSDs of 3.2-17.2 %), detection limits (DL) and limit of detections (LODs) was in the range of DL = 0.53-1.63 ppb and LOD = 0.19-2.51 ppb for pure acid standards, and DL = 0.18-3.41 ppb and LOD = 0.28-5.46 ppb for matrix matched acid standards. Finally, the method was validated on the acidic fraction of a mixed anion-exchange SPE of oil polluted water. Thousands of degradation products from parent alkylated polycyclic aromatic hydrocarbons (PAHs) and aliphatic hydrocarbons, such as aliphatic acids and mono-, di- and tri- aromatic acids were analyzed by the applied method and compound groups were tentatively identified.

Effect of folate supplementation on insulin sensitivity and type 2 diabetes: a meta-analysis of randomized controlled trials.

Background: Various mechanisms link higher total homocysteine to higher insulin resistance (IR) and risk of type 2 diabetes (T2D). Folate supplementation is recognized as a way to lower homocysteine. However, randomized controlled trials (RCTs) show inconsistent results on IR and T2D outcomes. Objective: The aim of this study was to examine the effect of folate supplementation on IR and T2D outcomes. Design: We conducted a systematic literature search in PubMed, Web of Science, and EMBASE and prior systematic reviews and meta-analyses and identified 29 RCTs (22,250 participants) that assessed the effect of placebo-controlled folate supplementation alone or in combination with other B vitamins on fasting glucose, insulin, homeostasis model assessment for insulin resistance (HOMA-IR), glycated hemoglobin (HbA1c), or risk of T2D. The meta-analysis was conducted using both random- and fixed-effects models to calculate weighted mean differences (WMDs) or risk ratios with 95% CIs. Subgroup analyses were conducted based on intervention type (folate alone or in combination with other B vitamins), as well as analysis based on population characteristics, duration, dose, and change in homocysteine. Results: When compared with placebo, folate supplementation lowered fasting insulin (WMD: -13.47 pmol/L; 95% CI: -21.41, -5.53 pmol/L; P < 0.001) and HOMA-IR (WMD: -0.57 units; 95% CI: -0.76, -0.37 units; P < 0.0001), but no overall effects were observed for fasting glucose or HbA1c. Heterogeneity was low in all meta-analyses, and subgroup analysis showed no signs of effect modification except for change in homocysteine, with the most pronounced effects in trials with a change of >2.5 µmol/L. Changes in homocysteine after folate supplementation correlated with changes in fasting glucose (ß = 0.07; 95% CI: 0.01, 0.14; P = 0.025) and HbA1c (ß = 0.46; 95% CI: 0.06, 0.85; P = 0.02). Only 2 studies examined folate supplementation on risk of T2D, and they found no change in RR (pooled RR: 0.91; 95% CI: 0.80, 1.04; P = 0.16). Conclusion: Folate supplementation might be beneficial for glucose homeostasis and lowering IR, but at present there are insufficient data to conclusively determine the effect on development of T2D. This trial was registered on the Prospero database as CRD42016048254.

A review of the characteristics of dietary fibers relevant to appetite and energy intake outcomes in human intervention trials.

Background: Many intervention studies have tested the effect of dietary fibers (DFs) on appetite-related outcomes, with inconsistent results. However, DFs comprise a wide range of compounds with diverse properties, and the specific contribution of these to appetite control is not well characterized.Objective: The influence of specific DF characteristics [i.e., viscosity, gel-forming capacity, fermentability, or molecular weight (MW)] on appetite-related outcomes was assessed in healthy humans.Design: Controlled human intervention trials that tested the effects of well-characterized DFs on appetite ratings or energy intake were identified from a systematic search of literature. Studies were included only if they reported 1) DF name and origin and 2) data on viscosity, gelling properties, fermentability, or MW of the DF materials or DF-containing matrixes.Results: A high proportion of the potentially relevant literature was excluded because of lack of adequate DF characterization. In total, 49 articles that met these criteria were identified, which reported 90 comparisons of various DFs in foods, beverages, or supplements in acute or sustained-exposure trials. In 51 of the 90 comparisons, the DF-containing material of interest was efficacious for ≥1 appetite-related outcome. Reported differences in material viscosity, MW, or fermentability did not clearly correspond to differences in efficacy, whereas gel-forming DF sources were consistently efficacious (but with very few comparisons).Conclusions: The overall inconsistent relations of DF properties with respect to efficacy may reflect variation in measurement methodology, nature of the DF preparation and matrix, and study designs. Methods of DF characterization, incorporation, and study design are too inconsistent to allow generalized conclusions about the effects of DF properties on appetite and preclude the development of reliable, predictive, structure-function relations. Improved standards for characterization and reporting of DF sources and DF-containing materials are strongly recommended for future studies on the effects of DF on human physiology. This trial was registered at http://www.crd.york.ac.uk/PROSPERO as CRD42015015336.

Supplementation with dairy calcium and/or flaxseed fibers in conjunction with orlistat augments fecal fat excretion without altering ratings of gastrointestinal comfort.

BACKGROUND: Orlistat is a lipase inhibitor which reduced absorption of dietary fat by ~30% thereby inducing a weight loss; however, side effects occur as a consequence of increased colonic fat content. To test the hypothesis that most gastrointestinal side events induced by treatment with orlistat could be prevented/ameliorated by concomitant use of natural dietary components, flaxseed fiber (FF) and/or dairy calcium (Ca), binding liquid fats to more solid complexes. METHODS: A randomized controlled dietary intervention study. Thirty-eight obese adults completed a 1-week run-in period, where all participants were treated with orlistat (60 mg t.i.d) and were hereafter randomized to 12 weeks dietary supplementation with/without 5 g FF (FF+/FF-) and/or 1200 mg dairy calcium (Ca+/Ca-) in conjunction with orlistat. All feces were collected for 3 days, and diet was recorded for 5 days, during run-in and week 4. The primary end-point, gastrointestinal symptoms, was assessed biweekly. At baseline and after 12 weeks, cardiometabolic risk markers and anthropometrics were evaluated as secondary end-points. RESULTS: Both FF and Ca increased fecal fat excretion (P = 0.02 and P = 0.04, respectively). Although fecal fat excretion increased by ~100% in the FF+/Ca + group, and only by ~12% in the FF-/Ca + group, no interaction between FF and Ca was present, suggesting an additive effect. The fecal fat excretion was ~10 g/d higher with FF and Ca (~25 g/d) compared to fecal fat excretion with orlistat alone (~15 g/d). Mean ratings of severity of diarrhea tended to increase with Ca (P = 0.03) but not with FF. No other gastrointestinal symptoms, or a composite score of symptoms, were affected by the dietary supplements. Body weight was reduced in all groups but did not differ between groups, whereas waist circumference was most reduced in the FF+/Ca + group. No effects of dietary supplements on cardiometabolic risk factors were observed, except a slight increase in diastolic blood pressure (P = 0.03) with FF, but not Ca. CONCLUSIONS: Our results do not support an improvement in orlistat-induced gastrointestinal side effects by concomitant use of FF and Ca. However, fecal fat excretion was increased with both FF and Ca in the absence of a worsening of symptoms, warranting further studies powered to detect potential additive weight loss effects. TRIAL REGISTRATION: Ethical Committee of the Capital Region of Denmark reg. no. H-1-2010-110, 02-11-2010 database no. NCT01320228, 21-03-2011.

Marine biodegradation of crude oil in temperate and Arctic water samples.

Despite increased interest in marine oil exploration in the Arctic, little is known about the fate of Arctic offshore oil pollution. Therefore, in the present study, we examine the oil degradation potential for an Arctic site (Disko Bay, Greenland) and discuss this in relation to a temperate site (North Sea, Denmark). Biodegradation was assessed following exposure to Oseberg Blend crude oil (100 mg L(-1)) in microcosms. Changes in oil hydrocarbon fingerprints of polycyclic aromatic hydrocarbons (PAHs), alkyl-substituted PAHs, dibenzothiophenes, n-alkanes and alkyltoluenes were measured by gas chromatography-mass spectrometry (GC-MS). In the Disko Bay sample, the degradation order was n-alkanes>alkyltoluenes (para->meta->ortho-isomers)>PAHs and dibenzothiophenes, whereas, the degradation order in the North Sea samples was PAHs and dibenzothiophenes>alkyltoluenes>n-alkanes. These differences in degradation patterns significantly affect the environmental risk of oil spills and emphasise the need to consider the specific environmental conditions when conducting risk assessments of Arctic oil pollution.

Linseed dietary fibers reduce apparent digestibility of energy and fat and weight gain in growing rats.

Dietary fibers (DF) may affect energy balance, an effect often ascribed to the viscous nature of some water soluble DF, which affect luminal viscosity and thus multiple physiological processes. We have tested the hypothesis that viscous linseed DF reduce apparent nutrient digestibility, and limit weight gain, in a randomized feeding trial where 60 male, growing, Wistar rats, with an initial weight of ~200 g, were fed different diets (n = 10 per group): low DF control (C), 5% DF from cellulose (5-CEL), CEL + 5% DF from whole (5-WL) or ground linseed (5-GL), CEL + 5% DF from linseed DF extract (5-LDF), and CEL + 10% DF from linseed DF extract (10-LDF). Diets were provided ad libitum for 21 days. Feed intake and faecal output were measured during days 17-21. Faecal fat excretion increased with increasing DF content and was highest in the 10-LDF group. Apparent fat digestibility was highest with the C diet (94.9% ± 0.8%) and lowest (74.3% ± 0.6%) with the 10-LDF diet, and decreased in a non-linear manner with increasing DF (p < 0.001). Apparent fat digestibility also decreased with increased accessibility of DF (5-WL vs. 5-GL) and when the proportion of viscous DF increased (5-GL vs. 5-LDF). The 10-LDF resulted in a lower final body weight (258 ± 6.2 g) compared to C (282 ± 5.9 g), 5-CEL (281 ± 5.9 g), and 5-WL (285 ± 5.9 g) (p < 0.05). The 10-LDF diet reduced body fat compared to 5-CEL (p < 0.01). In conclusion, DF extracted from linseed reduced apparent energy and fat digestibility and resulted in restriction of body weight gain in growing rats.

Plasma enterolactone and incidence of endometrial cancer in a case-cohort study of Danish women.

The phyto-oestrogen enterolactone has been hypothesised to protect against hormone-dependent cancers, probably through its antioestrogenic potential. We investigated whether a higher level of plasma enterolactone was associated with a lower incidence of endometrial cancer in a case-cohort study in the 'Diet, Cancer and Health' cohort. The cohort study included 29 875 women aged 50­64 years enrolled between 1993 and 1997. Information on diet and lifestyle was provided by self-administrated questionnaires and blood was drawn from each participant. Time-resolved fluoroimmunoassay was used for biochemical determination of plasma enterolactone. A total of 173 cases and 149 randomly selected cohort members were included. We estimated incidence rate ratio (IRR) and 95% CI by a Cox proportional hazards model. A 20 nmol/l higher plasma concentration of enterolactone was associated with a non-significant lower risk of endometrial cancer (IRR 0.93, 95% CI 0.84, 1.04). When excluding women with low enterolactone concentrations (quartile 1) due to potential recent antibiotic use, the association became slightly stronger, but remained non-significant (IRR 0.90, 95% CI 0.79, 1.02). Menopausal status, hormone replacement therapy or BMI did not modify the association. In conclusion, we found some support for a possible inverse association between plasma enterolactone concentration and endometrial cancer incidence.

Effect of alginate supplementation on weight loss in obese subjects completing a 12-wk energy-restricted diet: a randomized controlled trial.

BACKGROUND: Acute studies with alginate-based preloads suggested that these strong gelling fibers may induce increased feelings of satiety and reduce energy intakes. However, the long-term efficacy and safety of alginate supplementation on body weight regulation are lacking. OBJECTIVE: The primary aim of the study was to investigate the effects in subjects of alginate supplementation in conjunction with energy restriction (-300 kcal/d) on loss of body weight and fat and, second, on metabolic risk markers in comparison with in a placebo group. DESIGN: In a parallel, double-blind, placebo-controlled study, we randomly assigned 96 obese subjects to either an energy-restricted diet plus a placebo preload supplement or an energy-restricted diet plus an alginate-based preload supplement (15 g fiber). The preload was administered as a beverage 3 times/d before main meals for a period of 12 wk. RESULTS: No differences in loss of body weight and fat between groups were shown in the intension-to-treat (ITT) analysis (P > 0.1). However, in the completer analysis (n = 80), we showed a greater weight loss with alginate (6.78 ± 3.67 kg) than with the placebo (5.04 ± 3.40 kg) (P = 0.03), which was mainly attributed to a reduction in the percentage of body fat (P = 0.03). In the ITT analysis, a larger decrease in systolic and diastolic blood pressure was shown in the placebo group than in the alginate group (P < 0.05). Plasma concentrations of glucose, insulin, C-reactive protein, and ghrelin, HOMA-IR, and lipid metabolism did not differ between treatment groups in the ITT analysis (P > 0.1). CONCLUSION: These results suggest that alginate supplementation as an adjunct to energy restriction may improve weight loss in obese subjects who complete a 12-wk dietary intervention.

[Use of complementary and alternative medicine at Danish rehabilitation institutions and drug centres]. / Komplementær og alternativ medicin bruges på danske rehabiliterings- og rusmiddelinstitutioner.

The study has investigated the use of complementary and alternative medicine (CAM) at Danish rehabilitation institutions and drug centres. Questionnaires were sent to 147 centre leaders at 52 rehabilitation institutions and 95 drug centres. CAM is offered at 37.5% of the rehabilitation institutions and at 61.1% of the drug centres. Twenty-one different CAM modalities were used. Acupuncture and NADA-acupuncture had the highest prevalence. Motives for offering CAM were most often experienced effects within the institution and inspiration from other institutions.

Flaxseed dietary fiber supplements for suppression of appetite and food intake.

UNLABELLED: We conducted two single-blinded randomized crossover acute studies with 24 and 20 subjects, respectively, to compare (I) CONTROL vs. Flax drink; and (II) Flax drink vs. Flax tablets. The subjects were exposed to one of the treatments after an overnight fast, and rated appetite sensation for 120 min using visual analog scales (VAS). Hereafter they consumed an ad libitum early lunch to assess energy intake. The treatments were iso-caloric and iso-volumeric: CONTROL: 300 mL drink; Flax drink: CONTROL drink with addition flax fiber extract (2.5 g of soluble fibers); and Flax tablet: CONTROL drink with flax fiber tablets (2.5 g of soluble fibers). Flax drink increased sensation of satiety and fullness compared to CONTROL and a significant decrease in subsequent energy intake was observed after the Flax drink compared to CONTROL (2937 vs. 3214 kJ). Appetite ratings were similar for Flax tablets and Flax drink as they did not differ by more than 1-4%. Subsequent energy intake was similar after the two treatments (3370 vs. 3379 kJ). A small dose of flaxseed fiber significantly suppresses appetite and energy intake. Furthermore, flaxseed fibers administered as drinks or tablets produce similar responses.

Calcium from salmon and cod bone is well absorbed in young healthy men: a double-blinded randomised crossover design.

BACKGROUND: Calcium (Ca) - fortified foods are likely to play an important role in helping the consumer achieve an adequate Ca intake, especially for persons with a low intake of dairy products. Fish bones have a high Ca content, and huge quantities of this raw material are available as a by-product from the fish industry. Previously, emphasis has been on producing high quality products from fish by-products by use of bacterial proteases. However, documentation of the nutritional value of the enzymatically rinsed Ca-rich bone fraction remains unexplored. The objective of the present study was to assess the bioavailability of calcium in bones of Atlantic salmon (oily fish) and Atlantic cod (lean fish) in a double-blinded randomised crossover design. METHODS: Ca absorption was measured in 10 healthy young men using 47Ca whole body counting after ingestion of a test meal extrinsically labelled with the 47Ca isotope. The three test meals contained 800 mg of Ca from three different calcium sources: cod bones, salmon bones and control (CaCO3). RESULTS: Mean Ca absorption (+/- SEE) from the three different Ca sources were 21.9 +/- 1.7%, 22.5 +/- 1.7% and 27.4 +/- 1.8% for cod bones, salmon bones, and control (CaCO3), respectively. CONCLUSION: We conclude that bones from Atlantic salmon and Atlantic cod are suitable as natural Ca sources in e.g. functional foods or as supplements.

An onion byproduct affects plasma lipids in healthy rats.

Onion may contribute to the health effects associated with high fruit and vegetable consumption. A considerable amount of onion production ends up as waste that might find use in foods. Onion byproduct has not yet been explored for potential health benefits. The aim of this study is to elucidate the safety and potential role of onion byproducts in affecting risk markers of cardiovascular disease (CVD). For that purpose, the effects of an onion byproduct, Allium cepa L. cepa 'Recas' (OBP), and its two derived fractions, an ethanolic extract (OE) and a residue (OR), on the distribution of plasma lipids and on factors affecting cholesterol metabolism in healthy rats have been investigated. The OBP or its fractions did not significantly reduce cholesterol or down-regulate hepatic 3-hydroxy-3-methylglutaryl-coenzyme A reductase (Hmgcr) gene expression. The OR even had the effect of increasing plasma triacylglycerides (TAG) and cholesterol in the very low density lipoprotein (VLDL-C) fraction. Neither total bile acids nor total primary or secondary bile acids were significantly affected by feeding rats the OBP or its fractions. Principal component analysis combining all markers revealed that the controls could be completely separated from OBP, OE, and OR groups in the scores plot and also that OE and OR groups were separated. Plasma lipids and bile acid excretion were the discriminating loading factors for separating OE and OR but also contributed to the separation of onion-fed animals and controls. It was concluded that the onion byproduct did not present significant beneficial effects on individual markers related to plasma lipid transport in this healthy rat model but that onion byproduct contains factors with the ability to modulate plasma lipids and lipoprotein levels.

Six weeks phylloquinone supplementation produces undesirable effects on blood lipids with no changes in inflammatory and fibrinolytic markers in postmenopausal women.

BACKGROUND: Cardiovascular disease is the major cause of death in the Western world, but some recent studies indicate that vitamin K may play a role in atherosclerosis protection. AIM OF STUDY: The aim of this study was to evaluate the effect of phylloquinone supplementation on blood lipids, inflammatory markers and fibrinolytic activity in postmenopausal women. METHODS: Thirty-one postmenopausal women completed this placebo-controlled, randomized crossover study and received 500 microg phylloquinone or placebo in addition to their habitual diet during two periods of 6 weeks' duration. Blood concentration of lipids, inflammatory markers and fibrinolytic parameters were measured after each period. RESULTS: Inflammatory markers, fibrinolytic parameters, total cholesterol and LDL-C were unaffected by the supplementation, whereas a 15% increase was seen in triacylglycerols (P = 0.015) and a 5% decrease in HDL-C (P = 0.06). CONCLUSIONS: Six weeks supplementation with a dose of phylloquinone similar to that obtainable from the diet induced a deterioration of the lipid profile with no improvement in any of the other risk markers analysed. Thus, these results do not support a cardioprotective effect of vitamin K as has been suggested by others.

Effect of phylloquinone supplementation on biochemical markers of vitamin K status and bone turnover in postmenopausal women.

While current intakes of phylloquinone (vitamin K1) in many populations are believed to be sufficient to maintain normal blood coagulation, these may be insufficient to cover the requirements for optimal bone metabolism. Therefore, the objective of the present study was to investigate the effect of increasing phylloquinone intakes above the usual dietary intake for 6 weeks on biochemical markers of vitamin K status and bone turnover in postmenopausal women. Thirty-one postmenopausal women completed this 3 x 6-week randomised cross-over study, in which volunteers were supplemented with 0 (placebo), 200, and 500 microg phylloquinone/d. In addition, the volunteers were given 10 microg vitamin D3/d throughout the study period. With increasing phylloquinone intake, the concentration of serum gamma-carboxylated and under-gamma-carboxylated osteocalcin was significantly increased and decreased, respectively, in a dose-dependent manner (P < 0.001). Mean serum phylloquinone concentration was significantly (P < 0.001) higher with daily supplementation with 500 microg phylloquinone/d compared with that during either of the placebo or 200 microg phylloquinone/d supplementation periods, which did not differ (P = 0.15). Serum total osteocalcin was significantly (P < 0.001) increased in response to daily supplementation with 500 (but not 200) microg phylloquinone compared with placebo. Serum bone-specific alkaline phosphatase as well as the urinary markers of bone resorption (N-telopeptide cross-links of collagen, pyridinoline and deoxypyridinoline) and urinary gamma-carboxyglutamate were unaffected by phylloquinone supplementation. In conclusion, while daily supplementation with 200 and 500 microg phylloquinone/d for 6 weeks increased vitamin K status in postmenopausal women, it had no effect on bone turnover.

The importance of dietary composition for efficacy of iron absorption measured in a whole diet that includes rye bread fortified with ferrous fumerate: a radioisotope study in young women.

Fe absorption is affected by many dietary factors. The objective of the present study was to measure the effects of high v. low content of vitamin C, meat and phytic acid in whole diets with Fe-fortified bread on the efficacy of Fe absorption. Thirty-two healthy women with low Fe stores were randomised to three groups, each of which was given two of six test diets containing either low/high amounts of vitamin C, meat or phytic acid, respectively, in a cross-over design. Each diet was served throughout a 5 d period. Fe-fortified rye bread, extrinsically labelled with (59)Fe, was given with all main meals. Fe absorption was determined from whole-body counter measurements of (59)Fe retention. The fractional non-haem Fe absorption (corrected to a 40 % standard absorption by measurements from the reference dose) was 1.9 % v. 3.4 % (P=0.04) for the low/high vitamin C diets, 3.0 % v. 3.5 % (P=0.58) on the low/high meat diets and 4.9 % v. 3.8 % (P=0.24) on the low/high phytic acid diet, respectively. The total Fe absorbed (geometric mean with standard error) varied from 0.43 (se 0.11) mg from the diet with lowest bioavailability to 1.09 (se 0.18) mg from the diet with highest bioavailability (P<0.001). The present whole-diet study indicates that diet composition is a strong predictor of Fe absorption. In the diet with a low content of enhancers and a high content of inhibitors, vitamin C improved non-haem Fe absorption. The total Fe absorption varied 2.5-fold after small alterations of the content of enhancers and inhibitors in the diet.

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.