Vitamin E plasma kinetics in swine show low bioavailability and short half-life of -α-tocopheryl acetate.

Vitamin E is important for animal production because of its effects on health and product quality, but the amount and form required remains controversial. Our objective was to quantify the absolute bioavailability of oral -α-tocopheryl acetate (α-TAc) in swine (22 ± 1 kg and 8 wk old, fitted with jugular catheters) adapted to a diet supplemented with 75 mg/kg -α-TAc; 75 mg/kg was chosen because this level represents the nonweighted average inclusion level in piglet diets across Western key swine-producing countries. For this, a 350-g test meal (6% fat) was supplied at time 0 containing 75 mg deuterated (D9) -α-TAc to 9 animals, and 8 animals received an intravenous () dose containing deuterated (D6) RRR-α-tocopherol (α-T) at one-eighth the oral dose and a test meal without supplemental vitamin E. Plasma samples (12 to 13 per animal) were obtained at incremental intervals over 75 h for analysis of deuterated α-T using liquid chromatography-tandem mass spectrometry. Surprisingly, the i.v. dose rapidly disappeared from plasma and then reappeared. The half-life for this first peak was only 1.7 ± 0.3 min. The second peak had an appearance rate (Ka) of 0.10 ± 0.06 d and a half-life of 5.9 ± 1.2 h. Oral dosing resulted, after a lag of 56 min, in a Ka of 0.91 ± 0.21 d and a half-life of 2.6 ± 0.8 h. The bioavailability for oral α-TAc was 12.5%, whereas the area under the curve was only 5.4%. This low bioavailability, small area under the curve, and short half-life are likely because of various factors, that is, the use of only 6% fat in the diet, the use of the acetate ester and , and the high dose relative to requirements. In conclusion, i.v. dosed vitamin E shows both a rapid and a very slow pool, whereas orally dosed vitamin E shows a single slow pool. The oral material has a very short half-live (44% of i.v. or 2.6 h), low bioavailability (12.5%), and a very small area under the curve (5.4%), bringing into question the efficacy of typical doses of vitamin E in swine diets for alleviating oxidative stress.

Potential risk indicators of retained placenta and other diseases in multiparous cows.

Retained placenta (RP), defined as fetal membranes not being expelled within 24 h after calving, is a costly disease in multiparous dairy cows that has been linked to immune suppression, infections, elevated lipid mobilization, and depleted status of antioxidants including α-tocopherol, and that increases the risk of other diseases (OD) in early lactation. Early detection of cows at increased risk of developing RP, OD, or both in early lactation could improve treatment success and result in improved milk production and reproductive performance. To identify risk indicators of RP, OD, or both, we used a nested case-control design and compared multiparous dairy cows that developed RP (n=32) with cows that remained healthy (H; n=32) or cows that developed OD (n=32) in early lactation. We compared peripartal body condition score (BCS) as well as serum concentrations of α-tocopherol, metabolites [ß-hydroxybutyrate (BHBA), cholesterol, glucose, nonesterified fatty acids (NEFA), and urea N], haptoglobin, and macrominerals (i.e., calcium, magnesium, and phosphorus) on d -21, -14, -7, -3, -1, 0, 1, 3, 7, 14, 21, 28, 35, 42, and 49 postpartum. In addition, average serum concentrations were calculated for each cow for the last 3 wk prepartum, for 3 and 2 wk prepartum combined, for the last week prepartum, and for the morning after calving and compared between groups. The RP cows had lower BCS than the H or OD cows until 2 wk postpartum. During the prepartal periods, RP and OD cows had lower α-tocopherol concentrations (corrected or not for cholesterol concentration) and higher NEFA and BHBA concentrations than H cows. Thus, lower prepartal BCS could be an early predictor for RP risk, and lower α-tocopherol concentrations and higher NEFA and BHBA concentrations could be early predictors for disease.

Depleted serum vitamin E concentrations precede left displaced abomasum in early-lactation dairy cows.

Cows with left displaced abomasum (LDA), a costly disease occurring primarily in multiparous dairy cows during early lactation, have been reported to have 40% lower circulating concentrations of vitamin E. It is unknown, however, whether the lower circulating α-tocopherol concentrations precede LDA or remain after LDA. Using a nested case-control design, blood samples taken at d -21, -14, -7, -3, -1, 0, 1, 3, 7, 14, 21, 28, 35, 42, and 49 postpartum from 7 multiparous Holstein cows diagnosed with LDA between d 6 and 32 postpartum and 10 healthy Holstein cows from the same herd were analyzed for serum concentrations of α-tocopherol and indicators of energy and nutrient status and inflammation. In addition to indicators of negative energy balance and inflammation, lower serum α-tocopherol concentrations preceded LDA and persisted after LDA correction. At the last blood sampling before LDA diagnosis, cows had serum α-tocopherol concentrations 45% lower (5.0 ± 0.9 vs. 9.1 ± 0.9 µM) and α-tocopherol to cholesterol molar ratios 39% lower (1.90 ± 0.19 vs. 3.09 ± 0.26) than those of healthy cows. Serum α-tocopherol concentrations remained lower (<10 vs. ~15 µM) up to d 49 postpartum in cows that had LDA. These findings indicate that lower serum α-tocopherol concentrations are a potential early indicator for the development of LDA in multiparous cows.

Dietary intake associated with serum versus urinary carboxymethyl-lysine, a major advanced glycation end product, in adults: the Energetics Study.

BACKGROUND/OBJECTIVES: Advanced glycation end products (AGEs) are implicated in the pathogenesis of atherosclerosis, diabetes and kidney disease. The objective was to describe dietary intake, the dominant source of exposure to AGEs, with carboxymethyl-lysine (CML), a major AGE, in serum and urine, respectively. SUBJECTS/METHODS: Serum and urinary CML were measured in 261 adults, aged 21-69 years, and compared with diet as assessed by six separate 24-h dietary recalls. RESULTS: Median (25th, 75th percentile) serum and urinary CML concentrations were 686 (598, 803) µg/l and 1023 (812, 1238) µg/gm creatinine. There was no correlation between serum and urinary CML (r=-0.02, P=0.78). Serum CML was positively correlated with intake of soy, fruit juice, cold breakfast cereal, non-fat milk, whole grains, fruit, non-starchy vegetables and legumes, and negatively correlated with intake of red meat. Intake of fast food was not significantly correlated with serum CML. Urinary CML was positively correlated with intake of starchy vegetables, whole grains, sweets, nuts/seeds and chicken, and negatively correlated with intake of fast foods. Intake of AGE-rich foods such as fried chicken, French fries, bacon/sausage and crispy snacks were not significantly correlated with serum or urinary CML, except for a significant negative correlation between fried chicken and serum CML. CONCLUSIONS: These findings suggest that the high consumption of foods considered high in CML is not a major determinant of either serum or urinary CML. Further work is needed to understand the relationship of AGEs in blood and urine with the metabolism of dietary AGEs.

Nutrient biomarker patterns, cognitive function, and MRI measures of brain aging.

OBJECTIVE: To examine the cross-sectional relationship between nutrient status and psychometric and imaging indices of brain health in dementia-free elders. METHODS: Thirty plasma biomarkers of diet were assayed in the Oregon Brain Aging Study cohort (n = 104). Principal component analysis constructed nutrient biomarker patterns (NBPs) and regression models assessed the relationship of these with cognitive and MRI outcomes. RESULTS: Mean age was 87 ± 10 years and 62% of subjects were female. Two NBPs associated with more favorable cognitive and MRI measures: one high in plasma vitamins B (B1, B2, B6, folate, and B12), C, D, and E, and another high in plasma marine ω-3 fatty acids. A third pattern characterized by high trans fat was associated with less favorable cognitive function and less total cerebral brain volume. Depression attenuated the relationship between the marine ω-3 pattern and white matter hyperintensity volume. CONCLUSION: Distinct nutrient biomarker patterns detected in plasma are interpretable and account for a significant degree of variance in both cognitive function and brain volume. Objective and multivariate approaches to the study of nutrition in brain health warrant further study. These findings should be confirmed in a separate population.

Early feeding and dietary lipids affect broiler tissue fatty acids, vitamin E status, and cyclooxygenase-2 protein expression upon lipopolysaccharide challenge.

Newly hatched chicks are often subjected to delayed access to feed and water because of shipment distances and hatchery practices, which may reduce growth and development of the immune system. The current study investigated the effects of early vs. late access to feed and dietary lipids (n-3 vs. n-6) on lipopolysaccharide (LPS)-induced alterations in tissue fatty acids, vitamin E status, and cyclooxygenase-2 (COX-2) protein expression. The chicks (n = 16/group) were fed a high or low n-3 diet within 5 to 5 h 30 min (early) or after 48 h (late) of hatching. Feeding high n-3 diets increased eicosapentaenoic acid (EPA, 20:5 n-3), docosapentaenoic acid (22:5 n-3), and docosahexaenoic acid (DHA, 22:6 n-3) in the liver, spleen, and plasma (P < 0.05). Feeding low n-3 diets increased arachidonic acid in the liver and plasma (P < 0.05). Early access to feed led to increases in liver oleic acid and reduction in arachidonic acid as compared with late-fed birds (P < 0.05). No effect of time of feeding on fatty acids in the spleen was observed. Early feeding led to significant increases in linoleic and arachidonic acids in the plasma (P < 0.05). Stearic acid was higher in the plasma of low n-3 early-fed as opposed to low n-3 late-fed birds (P < 0.05). The LPS challenge led to an increase in liver total fat content (P < 0.05). The total fat content in the spleen and plasma were not affected by LPS injection (P > 0.05). The LPS-injected birds had decreases in oleic acid in the liver and plasma as compared with saline-injected birds (P < 0.05). Stearic acid increased upon LPS injection in the spleen and plasma (P < 0.05). Liver vitamin E content was significantly higher in saline-injected birds from the early high n-3 group compared with all treatment groups, except for the late low n-3 saline-injected birds (P < 0.05). Plasma vitamin E was highest in the early low n-3 LPS-injected birds compared with all other treatment groups (P < 0.05). The COX2:actin ratio in the early high n-3 LPS-injected birds was higher than that of the saline-injected birds of the same treatment (P < 0.05). However, no difference in COX-2 expression was observed between LPS- or saline-injected fed early low n-3, late high n-3, or late low n-3 diets (P > 0.05). No effect of diet, time of feeding, or LPS challenge on plasma isoprostanes was observed (P > 0.05). These results suggest that dietary and management strategies directed at modulating tissue polyunsaturated fatty acid status may offer the promise of modulating lipid metabolism and COX-2 expression in commercial poultry.

Conjugated linoleic acid and fish oil in laying hen diets: effects on egg fatty acids, thiobarbituric acid reactive substances, and tocopherols during storage.

The effects of incorporating conjugated linoleic acid (CLA) and fish oil in laying hen diets on egg CLA, n-3 fatty acid, tocopherol, and TBA reactive substances (TBARS) during 60 d of storage were investigated. Hens were fed corn-soybean meal-based diets containing 3% yellow grease (YG), 2.75% yellow grease + 0.25% CLA (YG-CLA), 2.5% yellow grease + 0.25% CLA + 0.25% fish oil (YG-CLA-FO), or 2.75% yellow grease + 0.25% fish oil (YG-FO). Eggs were collected and stored at 4 degrees C up to 60 d. On storage d 0, 20, 40, and 60, eggs (n = 8) from each treatment were selected randomly, and tocopherol and TBARS contents were measured. Egg total lipid and fatty acids were determined on d 0 and 60 of storage. Feeding YG-CLA-FO led to a 5.4 and 7.7% reduction in egg total lipids on d 0 and 60 (P < 0.05) when compared with YG eggs. The YG-CLA and YG-CLA-FO diets led to a 12% increase in egg saturated fatty acids compared with YG eggs. The content of monounsaturated fatty acids were lower ( > 19%) in YG-CLA and YG-CLA-FO compared with YG. Egg n-3 was highest in YG-FO eggs and lowest in YG eggs (P < 0.0001). Storage over 60 d led to a 20 and 67% depletion of CLA in the YG-CLA and YG-CLA-FO eggs (P < 0.0001). A 29% reduction was observed in the total n-3 fatty acid content of YG-CLA-FO eggs at d 60 of storage when compared with d 0 of storage (P < 0.0001). Diet and storage increased TBARS (P < 0.0001), which was highest in YG-CLA eggs at 60 d of storage. The YG-CLA and YG-CLA-FO diets reduced alpha and gamma-tocopherol contents at all days of storage compared with YG eggs (P < 0.05). Regardless of diet, egg storage for 40 d or longer depleted egg tocopherol contents (P < 0.05). These data demonstrate that healthy eggs with increased n-3 fatty acids and CLA can be generated by minor diet modifications, but added tocopherol supplementation may be needed to reduce lipid peroxidation when n-3 or CLA is included in the hen diet.

Burn and smoke inhalation injury in sheep depletes vitamin E: kinetic studies using deuterated tocopherols.

To test the hypothesis that burn and smoke injury will deplete tissue alpha-tocopherol and cause its faster plasma disappearance, deuterium-labeled vitamin E was administered to sheep exposed to both surface skin burn and smoke insufflation, which cause injuries similar to those of human victims of fire accidents. Two different protocols were used: (1) deuterated vitamin E was administered orally with food at time 0 (just before injury) or (2) the labeled vitamin E was administered orally with food the day before injury. The animals, which had been operatively prepared seven days before, were anesthetized and then received both 40% body surface area third-degree burn and 48 breaths of cotton smoke or sham injuries. All were resuscitated with Ringer's lactate solution (4 ml/kg/% BSA burn/24 h) and mechanically ventilated. Blood samples were collected at various times after vitamin E dosing. In both studies the depletion of plasma alpha-tocopherol was faster in the injured sheep. The sheep given deuterated vitamin E 24 h before injury had similar maximum alpha-tocopherol concentrations at similar times. The exponential rates of alpha-tocopherol disappearance were 1.5 times greater and half-lives were 12 h shorter (p < 0.05) in the injured sheep. In separate studies, various tissues were obtained from sheep that were sacrificed from 4 to 48 h after injury. The liver alpha-tocopherol concentrations in sheep killed at various times after injury seem to show a linear decrease at a rate of 0.1 nmol alpha-tocopherol/g liver per hour, suggesting that the liver is supplying alpha-tocopherol to maintain the plasma and lung alpha-tocopherol concentrations, but that this injury is so severe the liver is unable to maintain lung alpha-tocopherol concentrations. These findings suggest that alpha-tocopherol should be administered to burn patients to prevent vitamin E depletion and to protect against oxidative stress from burn injury.

Relative bioactivity of dietary RRR- and all-rac-alpha-tocopheryl acetates in swine assessed with deuterium-labeled vitamin E.

This study evaluated the relative bioactivities of natural and synthetic stereoisomers of alpha-tocopherol in swine. Deuterium-labeled vitamin E (150 mg each of d3-RRR- [natural] and d6-all-rac- [synthetic] alpha-tocopheryl acetates) was administered orally to adult female pigs (n = 3) with the morning feed. Blood samples were obtained at 0, 3, 6, 9, 12, 36, 48, and 72 h after the dose. The time of maximum plasma d3-alpha-tocopherol concentration (0.486 microg/mL) occurred at 12 h, and d6-alpha-tocopherol peaked earlier (at 9 h) and at a lower (P < 0.05) concentration (0.288 microg/mL). The d3-/d6-alpha-tocopherol ratio increased from 1.35 (SD = 0.73) at 3 h after dosing to 2.0 (SD = 0.14) at 72 h (P = 0.03). The plasma disappearance rates of d3- and d6-alpha-tocopherols (post-maximum concentrations) were similar and were estimated to be 0.013 microg/mL per hour. In summary, swine discriminated between RRR- and all-rac-alpha-tocopherols, which resulted in an approximately twofold higher plasma alpha-tocopherol concentration arising from the RRR-form. This 2:1 ratio of RRR- to all-rac- is higher than the currently accepted USP definition of RRR-:all-rac- of 1.36:1.00.

Vitamin E kinetics in smokers and nonsmokers.

Does cigarette smoking increase vitamin E utilization in vivo? A trial was carried out in 6 smokers and 5 nonsmokers of comparable ages and serum lipids. Subjects consumed 75 mg each d(3)-RRR and d(6)-all rac-alpha-tocopheryl acetates (natural and synthetic vitamin E, respectively) daily for 7 d with a standardized breakfast. Fasting blood samples were drawn on days -7, -6, -5, -4, -3, -2, -1, 0, 1, 2, 3, 4, 5, 6, 7, 9, 14, 21 (negative days indicate supplementation). In both groups, plasma d(3)-alpha-tocopherol concentrations were approximately double of d(6)-alpha-tocopherol. At day 0, the %d(3) alpha-tocopherols (d(3)-alpha-tocopherol/total-alpha-tocopherol x 100) were similar in both smokers and nonsmokers. Subsequently, there was a trend toward a faster exponential disappearance of the plasma %d(3) alpha-tocopherol in smokers compared with nonsmokers (0.30 +/- 0.04 compared with 0.24 +/- 0.05, p =.0565). The calculated %d(3) half-lives were 55.6 +/- 7.4 h in smokers and 72.1 +/- 17.3 h in nonsmokers (p =.0630). By day 21, the %d(3) in smokers had decreased to 1.4% +/- 0.3% while it was 2.2% +/- 0.7% (p =.0418) in the nonsmokers. These data suggest that smoking increases plasma vitamin E disappearance, but further studies are needed to confirm this finding and to assess its cause.

Vitamin E kinetics and the function of tocopherol regulatory proteins.

Plasma and tissue alpha-tocopherol concentrations are remarkably stable, which suggests that they are regulated. alpha-Tocopherol transfer protein, tocopherol-associated protein, and tocopherol-binding protein bind alpha-tocopherol. These proteins might function as tocopherol regulatory proteins, although only tocopherol transfer protein has been shown to influence plasma and tissue alpha-tocopherol concentrations. Tissue alpha-tocopherol concentrations likely depend on tocopherol regulatory protein function and tissue lipid content, vitamin E uptake and efflux, oxidative stress, and interactions between vitamin E and other antioxidants. Pharmacokinetic models often divide tissues into rapidly perfused, slowly perfused, and very slowly perfused compartments. Tissue vitamin E concentrations might equilibrate more rapidly in tissues with greater perfusion, greater vitamin E uptake, increased amounts or activities of tocopherol regulatory protein, and lower lipid contents. The rate at which tissue concentrations approach equilibrium, however, does not predict the final equilibrium concentrations because of redistribution among tissues. Redistribution of vitamin E to adipose tissue from other tissues may be significant. Intracellular trafficking of vitamin E might occur in conjunction with membrane recycling because membrane constituents rapidly recycle between the plasma membrane and intracellular endocytic compartments. Thus, tocopherol regulatory proteins may modulate rather than directly regulate vitamin E tissue distribution and intracellular trafficking.

Oxidative stress in athletes during extreme endurance exercise.

Despite the many known health benefits of exercise, there is a body of evidence suggesting that endurance exercise is associated with oxidative stress. To determine whether extreme endurance exercise induces lipid peroxidation, 11 athletes (3 females, 8 males) were studied during a 50 km ultramarathon (trial 1) and during a sedentary protocol (trial 2) 1 month later. The evening before each trial, with dinner, subjects consumed 75 mg each d(3)-RRR and d(6)-all rac-alpha-tocopheryl acetates. Blood was obtained at baseline, 30 min pre-race, mid-race, post-race, 1 h post-race, 24 h post-race, and at corresponding times during trial 2. All 11 subjects completed the race; average run time was 391 +/- 23 min. Plasma F(2)-isoprostanes increased from 75 +/- 7 pg/ml at pre-race to 131 +/- 17 (p <.02) at post-race, then returned to baseline at 24 h post-race; F(2)-isoprostanes were unchanged during trial 2. Deuterated alpha-tocopherol disappearance rates were faster (2.8 x 10(-4) +/- 0.2 x 10(-4)) during the race compared to the sedentary trial (2.3 x 10(-4) +/- 0.2 x 10(-4); p <.03). These data suggest that extreme endurance exercise results in the generation of lipid peroxidation with a concomitant increase in vitamin E disappearance.

Quantitative analysis by liquid chromatography-tandem mass spectrometry of deuterium-labeled and unlabeled vitamin E in biological samples.

A method for quantification of unlabeled alpha-tocopherol and the deuterated tocopherols, RRR-alpha-5-(CD(3))-tocopherol (d(3)RRR) and all rac-alpha-5,7-(CD(3))(2) tocopherol (d(6)all-rac) in plasma by HPLC-tandem mass spectrometry (LC-MS/MS) has been developed. Deuterated and unlabeled alpha-tocopherols were separated by HPLC and were detected by positive ion multiple-reaction monitoring using a triple-quadrupole mass spectrometer equipped with a heated nebulizer-atmospheric pressure chemical ionization interface, following routine extraction of vitamin E from plasma. The accuracy and precision were evaluated by replicate analysis of standards and samples. Human plasma samples, which were obtained at different times after the subject had consumed a capsule containing 1:1 ratio of d(3)RRR and d(6)all rac-alpha-tocopheryl acetates, were analyzed with this method. Plasma deuterated alpha-tocopherols measured by LC-MS/MS followed the same pattern as previously demonstrated by GC-MS measurement, without requiring an extra derivitization step. The detection limit was 10 pmol for each form of alpha-tocopherol injected.

Alpha- and gamma-tocotrienols are metabolized to carboxyethyl-hydroxychroman derivatives and excreted in human urine.

Limited information is available regarding metabolism of vitamin E forms, especially the tocotrienols. Carboxyethyl-hydroxychromans (alpha- and gamma-CEHC) are human urinary metabolites of alpha- and gamma-tocopherols, respectively. To evaluate whether tocotrienols are also metabolized and excreted as urinary CEHC, urine was monitored following tocotrienol supplementation. Complete (24 h) urine collections were obtained for 2 d prior to (baseline), the day of, and 2 d after human subjects (n = 6) ingested tocotrienol supplements. The subjects consumed 125 mg gamma-tocotrienyl acetate the first week, then the next week 500 mg; then 125 mg alpha-tocotrienyl acetate was administered the third week, followed by 500 mg the fourth week. Urinary alpha- and gamma-CEHC were measured by high-performance liquid chromatography with electrochemical detection. Urinary gamma-CEHC levels rose about four- to sixfold in response to the two doses of gamma-tocotrienol and then returned to baseline the following day. Significant (P < 0.0001) increases in urinary alpha-CEHC were observed only following ingestion of 500 mg alpha-tocotrienyl acetate. Typically, 1-2% of alpha-tocotrienyl acetates or 4-6% of gamma-tocotrienyl acetates were recovered as their respective urinary CEHC metabolites. A gamma-CEHC excretion time course showed an increase in urinary gamma-CEHC at 6 h and a peak at 9 h following ingestion of 125 mg gamma-tocotrienyl acetate. In summary, tocotrienols, like tocopherols, are metabolized to CEHC; however, the quantities excreted in human urine are small in relation to dose size.

Removal of fat from cow's milk decreases the vitamin E contents of the resulting dairy products.

The present study was undertaken to determine whether decreases in fat contents result in lower vitamin E contents. Milk samples of varying fat contents (half and half, whole milk, reduced-fat milk, low-fat milk, and nonfat milk) were obtained from a local dairy on six different occasions. alpha-Tocopherol was the major form of vitamin E (>85%); gamma-tocopherol and alpha-tocotrienol were present to a lesser extent. As the fat contents of milk products decreased from 11 to 0.3%, the vitamin E contents decreased. For example, raw milk as compared to nonfat milk had both higher (-tocopherol contents (45.5 +/- 4.6 vs. 4.5 +/- 0.5 microg/100 g; P < or = 0.0001) and higher total lipids (3.46 +/- 0.49 vs. 0.30 +/- 0.07 g/100 g; P < or = 0.0001). Vitamin E, cholesterol, and total lipids increased as cream was added back to nonfat milk during production. For every 1 mg cholesterol increase, there was an increase of approximately 4 microg of alpha-tocopherol; for every 1 g total lipids increase, the alpha-tocopherol content increased by 17 microg. These data demonstrate that removal of milk fat markedly decreases the vitamin E content of various milk products.

Does vitamin E decrease heart attack risk? summary and implications with respect to dietary recommendations.

The hypothesis that oxidative stress has a role in atherosclerosis rests on a large body of experimental work carried out in animal models of heart disease. The situation is more complex in humans, in that the results from vitamin E supplementation trials have been conflicting. Nonetheless, there is emerging information that alpha-tocopherol may play a critical role in maintaining the function of key cellular components in the atherosclerotic process through its ability to inhibit the activity of protein kinase C, a key player in many signal transduction pathways. alpha-Tocopherol modulates pathways of platelet aggregation, endothelial cell nitric oxide production, monocyte/macrophage superoxide production and smooth muscle cell proliferation. Regulation of adhesion molecule expression and inflammatory cell cytokine production by alpha-tocopherol has also been reported. More studies are required to relate alpha-tocopherol intakes to optimal tissue responses in humans.

Cu2+ -induced low density lipoprotein peroxidation is dependent on the initial O2 concentration: an O2 consumption study.

Atherosclerotic plaques form in the arterial intima, where low density lipoprotein (LDL) is thought to be oxidatively modified at sites which may contain catalytic amounts of copper in the presence of low O2 tension. We have investigated O2 consumption during LDL peroxidation induced by Cu2+ ions in vitro and found two phases: a lag phase followed by a phase of rapid O2 consumption. The length of the lag phase was dependent on Cu2+ and on initial O2 concentrations; increasing either decreased the lag time; however, LDL. concentration had no effect. LDL-induced Cu2+ reduction, however, was not affected by low initial O2 concentrations, suggesting that O2 is not required for LDL-mediated reduction of Cu2+. Following the lag phase, O2 consumption was dependent upon LDL or initial O2 concentrations; Cu2+ concentrations had little effect, suggesting that the propagation phase is more dependent on the presence of LDL lipids and O2 as substrates for the reaction. In summary, LDL peroxidation takes place in the presence of Cu2+ at low O2 tension; however, the reaction is dependent upon initial O2 concentrations; increases shorten the lag phase and accelerate O2 consumption.

Dependence of plasma alpha-tocopherol flux on very low-density triglyceride clearance in humans.

To evaluate the effect of dietary fat-induced alterations in triglyceride (TG) metabolism on plasma and very low-density lipoprotein (VLDL)-alpha-tocopherol, nine healthy males (mean +/- SEM, age: 36 +/- 3 years, BMI: 24.7 +/- 1.1) consumed a 35%-fat diet (control) for one week followed by a 15% low-fat, high-carbohydrate diet for 5 weeks. After each dietary phase, the subjects ingested an evening meal along with a 50 mg capsule of (2)H(6)-RRR-alpha-tocopheryl acetate; blood samples were drawn over a 24 h period while the subjects remained fasted. Low-fat feeding increased fasting plasma TG concentrations by 53% (116 +/- 27 to 178 +/- 32, mg/dl, p < 0.0001) primarily by reducing VLDL-TG clearance. Total plasma alpha-tocopherol concentrations (labeled + unlabeled) were unchanged (25.8 +/- 2.3 vs. 26.4 +/- 3.0 nmol/ml plasma) and no differences between the diets were observed for plasma (2)H(6)-alpha-tocopherol concentration (4.8 +/- 0.6 nmol/ml, for both diets) or enrichments (18.1 +/- 1.8% average for both diets). However, low-fat feeding significantly increased the amount of alpha-tocopherol in the VLDL fraction (43%, p = 0.04) in concert with elevations in VLDL-apoB and TG. The alpha-tocopherol and TG content of VLDL varied in parallel in individual subjects and fractional replacement rates and clearance of alpha-tocopherol and TG in VLDL were closely correlated. Kinetic parameters were decreased by 32-39% from high-fat to low-fat. These data suggest that vitamin E bioavailability is similar between a 15 and 35% fat diet, with a redistribution of alpha-tocopherol in lipoproteins occurring during low-fat feeding (increased in the VLDL fraction, reduced in the other lipoproteins), and transfer of alpha-tocopherol from VLDL depends upon TG removal from the particle, consistent with previous observations in vitro and in animal studies.

Increased atherosclerosis in hyperlipidemic mice deficient in alpha -tocopherol transfer protein and vitamin E.

Although lipid peroxidation in the subendothelial space has been hypothesized to play a central role in atherogenesis, the role of vitamin E in preventing lipid peroxidation and lesion development remains uncertain. Here we show that in atherosclerosis-susceptible apolipoprotein E knockout mice, vitamin E deficiency caused by disruption of the alpha-tocopherol transfer protein gene (Ttpa) increased the severity of atherosclerotic lesions in the proximal aorta. The increase was associated with increased levels of isoprostanes, a marker of lipid peroxidation, in aortic tissue. These results show that vitamin E deficiency promotes atherosclerosis in a susceptible setting and support the hypothesis that lipid peroxidation contributes to lesion development. Ttpa(-/-) mice are a genetic model of vitamin E deficiency and should be valuable for studying other diseases in which oxidative stress is thought to play a role.