Phosphorus-31: A table-top method for 3D B1-field amplitude and phase measurements.

A novel method of high-spatial-resolution, 3D B1-field distribution measurements is presented. The method is independent of the MR-scanner, and it allows for automated acquisitions of complete maps of all magnetic field vector components for both proton and heteronuclear MR coils of arbitrary geometrical shapes. The advantage of the method proposed here, compared with methods based on measurements with an MR-scanner, is that a complete image of both receive and transmit B1-fields, including the phase of the B1-field, can be acquired. The B1 field maps obtained in this manner can be used for absolute quantification of metabolites in MRS experiments, as well as for intensity compensations in imaging experiments, both of which are important concepts in biological and medical MR applications. Another use might be in coil development and testing. A comparison with B1 field magnitude maps obtained with an MR-scanner was included to validate the accuracy of the proposed method.

Improving digit span assessment of short-term verbal memory.

We measured digit span (DS) in two experiments that used computerized presentation of randomized auditory digits with performance-adapted list length adjustment. A new mean span (MS) metric of DS was developed that showed reduced variance, improved test-retest reliability, and higher correlations with the results of other neuropsychological test results when compared to traditional DS measures. The MS metric also enhanced the sensitivity of forward versus backward span comparisons, enabled the development of normative performance criteria with subdigit precision, and elucidated changes in DS performance with age and education level. Computerized stimulus delivery and improved scoring metrics significantly enhance the precision of DS assessments of short-term verbal memory.

Dietary beta-carotene, vitamin C and E intake and breast cancer risk in the European Prospective Investigation into Cancer and Nutrition (EPIC).

So far, studies on dietary antioxidant intake, including beta-carotene, vitamin C and vitamin E, and breast cancer risk are inconclusive. Thus, we addressed this question in the European Prospective Investigation into Cancer and Nutrition. During a median follow-up time of 8.8 years, 7,502 primary invasive breast cancer cases were identified. Cox proportional hazard models were used to estimate hazard ratios (HR) and 95% confidence intervals (CI). All analyses were run stratified by menopausal status at recruitment and, additionally, by smoking status, alcohol intake, use of exogenous hormones and use of dietary supplements. In the multivariate analyses, dietary intake of beta-carotene, vitamin C and E was not associated with breast cancer risk in premenopausal [highest vs. lowest quintile: HR, 1.04 (95% CI, 0.85-1.27), 1.12 (0.92-1.36) and 1.11 (0.84-1.46), respectively] and postmenopausal women [0.93 (0.82-1.04), 0.98 (0.87-1.11) and 0.92 (0.77-1.11), respectively]. However, in postmenopausal women using exogenous hormones, high intake of beta-carotene [highest vs. lowest quintile; HR 0.79 (95% CI, 0.66-0.96), P (trend) 0.06] and vitamin C [0.88 (0.72-1.07), P (trend) 0.05] was associated with reduced breast cancer risk. In addition, dietary beta-carotene was associated with a decreased risk in postmenopausal women with high alcohol intake. Overall, dietary intake of beta-carotene, vitamin C and E was not related to breast cancer risk in neither pre- nor postmenopausal women. However, in subgroups of postmenopausal women, a weak protective effect between beta-carotene and vitamin E from food and breast cancer risk cannot be excluded.

Variation in intakes of calcium, phosphorus, magnesium, iron and potassium in 10 countries in the European Prospective Investigation into Cancer and Nutrition study.

BACKGROUND/OBJECTIVES: Adequate mineral intake is important for the maintenance of bone health, cellular function and general metabolism, and possibly in the aetiology of cancer and other chronic diseases. This study aimed at investigating variation in intakes of selected minerals across 10 European countries participating in the EPIC (European Prospective Investigation into Cancer and Nutrition) study. METHODS: Nutrient intakes for 36 034 subjects, aged between 35 and 74 years, in 27 centres were obtained using standardized 24-h dietary recall software (EPIC-SOFT). Mean intakes of calcium, phosphorus, magnesium, iron and potassium were calculated by centre and weighted by season and day of the week and were also stratified by age group. The contribution of food groups to total nutrient intake was calculated. RESULTS: There was clear geographical variability in intakes, with differences ranging from 35% for magnesium to 90% for iron in men and 36% for potassium to 75% for calcium in women, and a twofold difference in sources of haem iron (meat and fish). There was a geographical gradient in iron intake, with higher intakes in Southern than in Northern Europe and also around a twofold north-south gradient in the contribution of fruits and vegetables to potassium intake. Compared with reference intakes, the majority of age groups and centres had intakes above the recommended levels. Dairy foods and products contributed the most to calcium and phosphorus intake in almost all centres. Cereals and cereal products contributed the most to magnesium and iron intakes, except in Greece and Germany. CONCLUSIONS: Intakes of minerals vary substantially throughout Europe, with some geographical variability in their food sources.

Use of dietary supplements in the European Prospective Investigation into Cancer and Nutrition calibration study.

BACKGROUND: Dietary supplement use is increasing, but there are few comparable data on supplement intakes and how they affect the nutrition and health of European consumers. The aim of this study was to describe the use of dietary supplements in subsamples of the 10 countries participating in the European Prospective Investigation into Cancer and Nutrition (EPIC). METHODS: Specific questions on dietary supplement use were asked as a part of single 24-h recalls performed on 36,034 men and women aged 35-74 years from 1995 to 2000. RESULTS: Between countries, the mean percentage of dietary supplement use varied almost 10-fold among women and even more among men. There was a clear north-south gradient in use, with a higher consumption in northern countries. The lowest crude mean percentage of use was found in Greece (2.0% among men, 6.7% among women), and the highest was in Denmark (51.0% among men, 65.8% among women). Use was higher in women than in men. Vitamins, minerals or combinations of them were the predominant types of supplements reported, but there were striking differences between countries. CONCLUSIONS: This study indicates that there are wide variations in supplement use in Europe, which may affect individual and population nutrient intakes. The results underline the need to monitor consumption of dietary supplements in Europe, as well as to evaluate the risks and benefits.

Diet among breast cancer survivors and healthy women. The Norwegian Women and Cancer Study.

OBJECTIVE: To compare the diet and lifestyle in breast cancer survivors and healthy women. DESIGN: Cross-sectional study in the population-based Norwegian Women and Cancer cohort study, using a postal questionnaire on diet, lifestyle and health. SETTING: Nation-wide, population-based study. SUBJECTS: Women aged 41-70 years. Prevalent breast cancer cases (314 short-term with 1-5 years since diagnosis, 352 long-term with >5 years since diagnosis) were identified by linkage to the Norwegian Cancer Registry. The comparison group consisted of 54,314 women. INTERVENTIONS: Analyses of variance, with post hoc Bonferroni tests when significant differences were found. RESULTS: Overall there were few differences in the diet of the three groups. Short-term survivors ate more fruits and vegetables than healthy women (P<0.0001), and consumed more of nutrients associated with fruit and vegetables (fibre, mono- and disaccharides, folate, vitamin C and potassium). Short-term breast cancer survivors also had a higher use of dietary supplements and a lower level of physical activity, but did not differ from healthy women on other lifestyle factors. The long-time survivors did not differ from any of the other groups. CONCLUSION: Diet and lifestyle is generally similar between breast cancer survivors and healthy women, especially more than 5 years after diagnosis.

Dietary patterns and lifestyle factors in the Norwegian EPIC cohort: the Norwegian Women and Cancer (NOWAC) study.

OBJECTIVE: To identify different dietary patterns in Norway using a combination of cluster and factor analysis. DESIGN: Cross-sectional study. SETTING: Nation-wide, population-based study. SUBJECTS: The Norwegian EPIC cohort is a subcohort of the Norwegian Women and Cancer study (NOWAC), and consist 37.226 women aged 41-56 y who answered a food frequency questionnaire (FFQ) in 1998. INTERVENTIONS: The associations among 50 food variables were first investigated by using principal component analysis. Five important factors were found. The five principal components were then used as input in the cluster analysis. Different socioeconomic and lifestyle variables were examined. RESULTS: Six clusters of dietary patterns were found, and were labelled accordingly: 'traditional fish eaters', 'healthy eaters', 'average, less fish, less healthy', 'Western', 'traditional bread eaters', and 'alcohol users'. The traditional fish eaters and the traditional bread eaters were both highly represented in the north and west of Norway and were more likely to be present among persons with lower income and lower education. The healthy and the alcohol drinkers were found mostly in the south and east and were more likely to have higher income. Persons in the alcohol group were more likely to be current smokers. The western group had the highest percentage of three or more persons in the household and the shortest time since last birth, indicating that families with children dominate this group. CONCLUSION: Our data indicate six different dietary patterns in Norway, each with different socio-demographic and lifestyle characteristics. SPONSORSHIP: The Norwegian Cancer Society (E 04038/006).

Vitamin D status in a rural population of northern Norway with high fish liver consumption.

OBJECTIVE: To assess vitamin D status and the impact of three fish meals consisting of cod liver and fresh cod-liver oil on the plasma level of vitamin D metabolites in an area with high consumption of cod liver and cod-liver oil. DESIGN: Experimental field study. METHODS: Thirty-two volunteers from the Skjervøy (70 degrees N) municipality in northern Norway were recruited to consume three traditional mølje meals, consisting of cod, cod liver, fresh cod-liver oil and hard roe, in one week. The liver and fresh cod-liver oil consumed by the participants were weighed and recorded. Blood samples were collected before the first meal, and subsequently 12 h and 4 days after the last meal. The blood samples were analysed for the vitamin D metabolites 25-hydroxyvitamin D (25(OH)D) and 1,25-dihydroxyvitamin D (1,25(OH)2D). All participants answered a semi-quantitative food-frequency questionnaire, which was used to estimate usual daily nutrient intake. The study was carried out in the last part of March 2001. RESULTS: The median daily vitamin D intake estimated from the questionnaire was 9.9 microg. The proportion of subjects with baseline 25(OH)D level below 50 nmol l(-1) was 15.4% and none were below 37.5 nmol l(-1). Only "mølje consumption" and "time spent in daylight" were significantly associated with baseline log 25(OH)D. The mean total intake of vitamin D in the three servings was 272 microg (standard deviation 94 microg), ranging from 142 to 434 microg. Relative to baseline plasma concentration, the mean level of 25(OH)D decreased slightly in both post-consumption samples (P< or =0.03), while 1,25(OH)2D peaked 12 h after the final meal (P=0.03). CONCLUSION: Three mølje meals provided, on average, an amount of vitamin D equal to 54 times the recommended daily dose. Subjects with food consumption habits that included frequent mølje meals during the winter sustained satisfactory vitamin D levels in their blood, in spite of the long "vitamin D winter" (i.e. absence of ultraviolet-induced vitamin D production in the skin).

Predictors for cod-liver oil supplement use--the Norwegian Women and Cancer Study.

OBJECTIVE: To assess the use of cod-liver oil supplements among Norwegian women and to examine dietary, lifestyle, demographic, and health factors associated with use of this supplement. DESIGN: Cross-sectional study. SETTING AND SUBJECTS: The study is based on data from a food frequency questionnaire from 1998 answered by 37,226 women aged 41-55 y, who in 1991/1992 participated in the Norwegian component of the European Prospective Investigation into Cancer and Nutrition (EPIC). The Norwegian EPIC cohort was based on a random nation-wide sample of Norwegian women. RESULTS: Cod-liver oil supplement use was reported by 44.7% of the participating women. Subjects with higher education, high physical activity level, and body mass index (BMI) in the normal range were more likely to use cod-liver oil supplements. Consumption did also increase with increased age as well as with increased reported consumption of fruits, vegetables, fatty fish, lean fish, and vitamin D (excluding the vitamin D contribution from cod-liver oil). Energy intake was higher among cod-liver oil users than nonusers. Whole-year daily users of cod-liver oil were also more likely to take other dietary supplements (OR=2.45, 95% CI: 2.28-2.62). Never smokers were more likely to use cod-liver oil supplements than current smokers. CONCLUSION: Use of cod-liver oil is associated with several sociodemographic factors, self-reported health issues, and intake of fish, fruit, and vegetables. When assessing the relationship between cod-liver oil use and occurrence of chronic diseases potential confounders need to be considered. Cod-liver oil use seemed not to be matched with vitamin D needs. Thus, emphasis on assessing vitamin D status by measuring levels in blood should be investigated further, in particular, among people living in northern latitudes.

Muscle metabolism in fibromyalgia studied by P-31 magnetic resonance spectroscopy during aerobic and anaerobic exercise.

OBJECTIVE: To investigate mechanisms underlying the reduced work capacity of fibromyalgia (FM) patients were compared to healthy controls at specified workloads, using P-31 magnetic resonance spectroscopy (MRS). METHODS: The forearm flexor muscle group was examined with MRS at rest, at sub maximal and at maximal controlled dynamic work as well as at maximal isometric contraction. Aerobic fitness was determined by bicycle ergonometry. RESULTS: Metabolite concentrations and muscle pH were similar for patients and controls at lower workloads. At maximal dynamic and static contractions the concentration of inorganic phosphate was lower and at static contractions the pH decrease was smaller in patients. The performed work by patients was only 50% compared to controls and the patients experienced more pain. Maximal oxygen uptake was lower in the fibromyalgia group. Expired gas-analysis in this group showed ventilatory equivalents at similar relative levels of maximal work capacity. CONCLUSION: Fibromyalgia patients seem to utilise less of the energy rich phosphorous metabolites at maximal work despite pH reduction. They seemed to be less aerobic fitted and reached the anaerobic threshold earlier than the controls.

Uranium and thorium series radionuclides in drinking water from drilled bedrock wells: correlation to geology and bedrock radioactivity and dose estimation.

Natural radioactivity in drinking water from 328 drilled wells was studied in correlation to source parameters. Poor correlation to both aquifer geology and bedrock radioactivity was observed. Concentrations of 238U, 226Ra, 228Ra, 222Rn and 210Po in groundwater samples was in the ranges <0.027-5.3, <0.016-4.9, <0.014-1.24, 5-8105 and <0.05-0.947 Bq.l(-1) respectively. In about 80% of the sites the radon concentration exceeds the Nordic recommended exemption level for radon in drinking water and 15% of the sites exceed the action limit. The effective doses from ingestion were calculated and presented in association with geology. Doses due to ingestion ranged between 0.05 and 20.4 mSv.y(-1), where the average contribution from 222Rn amounted to 75%. In comparison, the effective doses from inhalation of indoor 222Rn ranged between 0.2 and 20 mSv.y(-1). The average contribution from inhalation of 222Rn in air to the total effective dose (ingestion+inhalation) was 58 +/- 22%, 73 +/- 18% and 77 +/- 16% (1 SD) for the age categories 1 y, 10 y and adults respectively.

27-hydroxycholesterol is an endogenous ligand for liver X receptor in cholesterol-loaded cells.

The nuclear receptors liver X receptor alpha (LXRalpha) (NR1H3) and LXRbeta (NR1H2) are important regulators of genes involved in lipid metabolism, including ABCA1, ABCG1, and sterol regulatory element-binding protein-1c (SREBP-1c). Although it has been demonstrated that oxysterols are LXR ligands, little is known about the identity of the physiological activators of these receptors. Here we confirm earlier studies demonstrating a dose-dependent induction of ABCA1 and ABCG1 in human monocyte-derived macrophages by cholesterol loading. In addition, we show that formation of 27-hydroxycholesterol and cholestenoic acid, products of CYP27 action on cholesterol, is dependent on the dose of cholesterol used to load the cells. Other proposed LXR ligands, including 20(S)-hydroxycholesterol, 22(R)-hydroxycholesterol, and 24(S),25-epoxycholesterol, could not be detected under these conditions. A role for CYP27 in regulation of cholesterol-induced genes was demonstrated by the following findings. 1) Introduction of CYP27 into HEK-293 cells conferred an induction of ABCG1 and SREBP-1c; 2) upon cholesterol loading, CYP27-expressing cells induce these genes to a greater extent than in control cells; 3) in CYP27-deficient human skin fibroblasts, the induction of ABCA1 in response to cholesterol loading was ablated; and 4) in a coactivator association assay, 27-hydroxycholesterol functionally activated LXR. We conclude that 27-hydroxylation of cholesterol is an important pathway for LXR activation in response to cholesterol overload.

Effects of cellular redox balance on induction of apoptosis by eicosapentaenoic acid in HT29 colorectal adenocarcinoma cells and rat colon in vivo.

BACKGROUND AND AIMS: Epidemiological evidence suggests n-3 polyunsaturated lipids may protect against colorectal neoplasia. Consumption of fish oil modulates crypt cytokinetics in humans, and crypt apoptosis in animal models. To explore these effects, we investigated involvement of caspase enzymes and cellular redox balance in the induction of apoptosis by eicosapentaenoic acid (EPA) in HT29 cells, and in rat colon in vivo. METHODS: Survival of HT29 cells grown with EPA in the presence of caspase inhibitors, antioxidants, or buthionine sulphoximine, an inhibitor of glutathione neosynthesis, was determined. The effects of EPA enriched fish oil and glutathione depletion on apoptosis in rat colon were assessed using microdissected crypts. RESULTS: Treatment of HT29 cells with EPA reduced viable cell number and activated caspase 3, prior to cell detachment. Antioxidants and caspase inhibitors blocked HT29 cell death whereas glutathione depletion increased it. Rats fed fish oil had higher crypt cell apoptosis than those fed corn oil, and glutathione depletion enhanced this effect. CONCLUSIONS: Incorporation of EPA into colonic epithelial cell lipids increases apoptosis. The results of this study, using both an animal and cell line model, support the hypothesis that this effect is mediated via cellular redox tone, and is sensitive to glutathione metabolism. The data suggest a mechanism whereby polyunsaturated fatty acids may influence the susceptibility of colorectal crypt cells to induction or progression of neoplasia.

Hyperlipidemia and reproductive failure in captive-reared alligators: vitamin E, vitamin A, plasma lipids, fatty acids, and steroid hormones.

Blood samples were collected from 26 captive-reared alligators (25 females; one male) and 12 (seven females and five males) wild "nuisance" alligators collected by wildlife personnel in south Louisiana in May 1995. The captive alligators, hatched from artificially incubated eggs in 1972-1973, had received vitamin E supplements during the 3 weeks before the blood sample was collected. Each sample was analyzed for vitamin E (alpha-tocopherol), vitamin A (retinol), total lipid, triacylglycerol, phospholipid, cholesterol, cholesteryl ester, free fatty acids, steroid hormones and a standard clinical blood panel. The fatty acid composition of the plasma lipid fraction was also analyzed. Results indicated that 18 of the captive females and three of the seven wild females were undergoing vitellogenesis, i.e. had elevated plasma estradiol and elevated plasma calcium. Vitellogenic females had higher vitamin E than non-vitellogenic females (77.4 microg/ml vs. 28.6 microg/ml in captive females; 24.0 microg/ml vs. 21 microg/ml in wild females). Plasma retinol was similar in all groups, ranging from 0.5 to 1.4 microg/ml and close to values reported in birds. All lipid fractions, with the exception of cholesteryl ester, were higher in captive alligators than in wild alligators. There were also significant differences in the fatty acid composition of wild and captive alligators. Plasma eicosapentaenoic and docasahexaenoic acid were higher in wild than in captive alligators, whereas linoleic was higher in captive than in wild.

Therapeutic synergy of TNP-470 and ionizing radiation: effects on tumor growth, vessel morphology, and angiogenesis in human glioblastoma multiforme xenografts.

We examined the effect on tumor growth, vessel morphology, and expression of angiogenic factors of combining radiotherapy and antiangiogenesis in the human glioblastoma line U87 grown in the flank or intracranially in the nude mouse. The antiangiogenic agent TNP-470 was given 6.7 mg/kg s.c. daily on day 1-7 starting 1 week after transplantation. Irradiation (IR), 10 Gy x 1, was administered on day 7. A series of tumors were excised 8 and 48 h after the end of treatment. The vascular morphology was evaluated in CD31 immunostained cryosections and by electron microscopy, and the pattern of expression of angiogenic factors (mRNA and protein) was quantitatively analyzed by phosphorimaging of Northern blots and Western blots. Significant inhibition of s.c. flank tumor growth relative to untreated controls was achieved by monotherapy with both TNP-470 (P < 0.001) and IR (P < 0.001). A significant enhancement of this effect was obtained by combining TNP-470 and IR (P < 0.05). We saw no effect of TNP-470 either alone or in addition to the effect of IR on the survival of mice with intracranial tumors. CD31 immunostaining of s.c. tumors showed acute endothelial swelling and luminal protrusion in irradiated tumor vessels but never in tumors pretreated with TNP-470, and not in the untreated controls. The vessel density (Chalkley point counts) was unchanged by TNP-470 therapy. In the TNP-470-treated tumors, we observed a distinct broadening of the endothelial basement membrane by an approximately 400-700-nm-thick electron-dense yet uncharacterized fibrillar material. TNP-470 treated tumors +/- IR also had a significantly increased mRNA expression of angiopoietin-1, whereas angiopoietin-2, vascular endothelial growth factor and basic fibroblast growth factor mRNA were unchanged by the treatments. In conclusion, TNP-470 significantly enhanced the tumor effect of ionizing IR, and our findings strongly indicate that acute microvascular damage after IR is effectively prevented by concurrent TNP-470 treatment. A significant up-regulation of angiopoietin-1 seems to play a role in this protective mechanism, which as yet is not fully elucidated.

Dietary n-3 PUFA increases the apoptotic response to 1,2-dimethylhydrazine, reduces mitosis and suppresses the induction of carcinogenesis in the rat colon.

The effect of dietary fish oil on colonic crypt cell apoptosis and proliferation was examined in male Wistar rats, 24 and 48 h after administration of 1,2-dimethylhydrazine (DMH), and its influence on the induction of aberrant crypt foci (ACF) in the distal colon was assessed. Rats (125-150 g) fed a high-fat semi-synthetic diet containing corn oil (CO) were given DMH (30 mg/kg body wt) or a sham injection of EDTA/NaCl. Animals were then fed either the CO diet or a diet in which fish oil (EPA 18.7%; DHA 8%) was substituted for corn oil. Subgroups of rats (n = 5) were killed after 24 and 48 h, and crypt cell apoptosis and proliferation were quantified by morphological criteria in isolated intact crypts from the mid and distal colon. Consumption of the fish oil diet (FO) was associated with increased apoptotic cell death (P < 0.001) and suppression of proliferation (P < 0.05) in colonic crypts both 24 and 48 h after DMH. In a second experiment, animals were given three injections of DMH or sham injections of carrier at weekly intervals. For 48 h after each injection animals were fed either the CO or FO diet, but otherwise maintained on the CO throughout. The number and crypt multiplicity of ACF in the distal colon were determined after 18 weeks, and animals given the FO diet for the 48 h period following carcinogen administration were found to have significantly fewer ACF than rats fed the CO diet (P < 0.05). The data demonstrate that the fatty acid composition of the diet is an important determinant in the induction of carcinogenesis by DMH. The proliferative and apoptotic response of the colonic crypt to carcinogen and fish oil, coupled with the reduced incidence of ACF, suggest n-3 PUFA can protect against the carcinogenic effects of DMH by mediating changes in the balance proliferation and cell death.

Oral ferrous sulfate supplements increase the free radical-generating capacity of feces from healthy volunteers.

BACKGROUND: Most dietary iron remains unabsorbed and hence may be available to participate in Fenton-driven free radical generation in conjunction with the colonic microflora, leading to the production of carcinogens or direct damage to colonocytes. OBJECTIVE: Our aims were to measure the proportion of fecal iron available to participate in free radical generation and to determine the effect of an oral supplement of ferrous sulfate on free radical generation. DESIGN: Eighteen healthy volunteers recorded their food intake and collected fecal samples before, during, and after 2 wk of supplementation (19 mg elemental Fe/d). Total, free, and weakly chelated fecal iron were measured and free radical production was determined by using an in vitro assay with dimethyl sulfoxide as a free radical trap. RESULTS: Fecal iron increased significantly during the period of supplementation and returned to baseline within 2 wk. The concentration of weakly bound iron in feces (approximately 1.3% of total fecal iron) increased from 60 micromol/L before to 300 micromol/L during supplementation, and the production of free radicals increased significantly (approximately 40%). Higher-carbohydrate diets were associated with reduced free radical generation. CONCLUSION: Unabsorbed dietary iron may increase free radical production in the colon to a level that could cause mucosal cell damage or increased production of carcinogens.

Effects of dietary fish oil supplementation on the phospholipid composition and fluidity of cell membranes from human volunteers.

BACKGROUND: Membrane fluidity is an important aspect of cellular physiology which may be manipulated by diet. METHODS: We studied the effect of dietary fish oil on the membrane composition of erythrocytes and cheek cells, and on membrane fluidity of erythrocytes as assessed by fluorescence recovery after photobleaching (FRAP). Healthy volunteers received a daily supplement of fish oil (930 mg EPA, 630 mg DHA) for 42 days. RESULTS: The intervention reduced the ratio of n-6 to total fatty acid in the phospholipid fraction of erythrocyte membranes but the n-3 fraction remained stable and the ratio of cholesterol to phospholipid increased. The level of EPA and DHA in cheek cells increased significantly during the intervention period. The mean diffusion coefficient of the fluorescent probe in erythrocyte membranes increased from 7.2 +/- 0.7 x 10-9 cm2/s at the start to 9.8 +/- 0.5 x 10-9 cm2/s after 21 days. Membrane fluidity remained higher than the initial value 42 days after withdrawal of the supplement. Fish oil also reduced platelet aggregation in response to ADP but there was no effect on plasma lipid profiles. CONCLUSION: We conclude that n-3 fatty acids influence erythrocyte membrane composition at relatively low levels of supplementation by a mechanism which does not necessarily involve an increase in the level of EPA or DHA present in the membrane.

cDNA cloning of mouse and human cholesterol 25-hydroxylases, polytopic membrane proteins that synthesize a potent oxysterol regulator of lipid metabolism.

Oxysterols regulate the expression of genes involved in cholesterol and lipid metabolism and serve as intermediates in cholesterol catabolism. Among the most potent of regulatory oxysterols is 25-hydroxycholesterol, whose biosynthetic enzyme has not yet been isolated. Here, we report the cloning of cholesterol 25-hydroxylase cDNAs from the mouse and human. The encoded enzymes are polytopic membrane proteins of 298 and 272 amino acids, respectively, which contain clusters of histidine residues that are essential for catalytic activity. Unlike most other sterol hydroxylases, cholesterol 25-hydroxylase is not a cytochrome P450, but rather it is a member of a small family of enzymes that utilize diiron cofactors to catalyze the hydroxylation of hydrophobic substrates. The cholesterol 25-hydroxylase gene lacks introns, and in the human it is located on chromosome 10q23. The murine gene is expressed at low levels in multiple tissues. Expression of cholesterol 25-hydroxylase in transfected cells reduces the biosynthesis of cholesterol from acetate and suppresses the cleavage of sterol regulatory element binding protein-1 and -2. The data suggest that cholesterol 25-hydroxylase has the capacity to play an important role in regulating lipid metabolism by synthesizing a co-repressor that blocks sterol regulatory element binding protein processing and ultimately leads to inhibition of gene transcription.