Antioxidant transport by the human placenta.

We investigated the transfer of three antioxidants - melatonin, S-adenosyl methionine (SAM) and various forms of vitamin E - across the term, normal human placenta. The transport technique involved the single, isolated placental cotyledon system in vitro. Melatonin crossed the placental rapidly, equally to the freely diffusible marker, antipyrine. There was no biotransformation of the agent. SAM was transferred slowly, similarly to passively transported L-glucose as a marker. There was a breakdown of SAM to at least one other derivative; the process appeared to be nonenzymatic. Vitamin E was transferred slowly, at a rate only 10% of L-glucose. The natural RRR (nonracemic) form of vitamin E was transported best. Free vitamin, rather than the acetate seems to be transferred best, a finding that will require further study.

Tocopherols and the etiology of colon cancer.

Colorectal cancer is the second most common cause of cancer deaths in the United States for both sexes. Considerable evidence suggests that the risk of this cancer is increased by the mutagenic actions of free radicals, which are produced during oxidation reactions. Dietary factors, the intestinal flora (bacteria), and endogenously produced metabolites contribute to the production of free radicals in the colon. Dietary antioxidants, such as vitamin E, should reduce the levels of these harmful oxidation products. In the absence of vitamin E, polyunsaturated fats can be oxidized in the colon to produce mutagens, such as lipid hydroperoxides and malondialdehyde. Furthermore, fecal bacteria can generate a high flux of reactive oxygen species (e.g., the superoxide radical [O2*-]) at the surface of the intestinal lumen, and inflammatory cells in close proximity to the colon can produce reactive nitrogen species (e.g., nitrogen dioxide [NO2]). Increasing evidence suggests that the different chemical (e.g., alpha- and gamma-tocopherol) and stereochemical (e.g., RRR- and all-racemic-alpha-tocopherol) forms of vitamin E have distinct biologic potencies, pharmacokinetics, and different abilities to prevent neoplastic transformation. This review considers and evaluates recent studies relating vitamin E and oxidative stress to colon cancer, emphasizing the distinct roles of alpha- and gamma-tocopherols. In addition, recent findings on the antioxidant/pro-oxidant status of the digesta (ingested food) are discussed with respect to the use of antioxidants in chemo-prevention trials for colon cancer.

Determinants of antioxidant status in humans.

Antioxidant status in humans reflects the dynamic balance between the antioxidant system and prooxidants and has been suggested as a useful tool in estimating the risk of oxidative damage. This paper reviews determinants of antioxidant status such as diet including antioxidant nutrient and nonnutrient intake, absorption and bioavailability, dietary components such as polyunsaturated fatty acids and transition metals, food storage and processing, chemical form, chirality and formulation of supplemental compounds and alcohol intake; environmental factors such as pollutants, ultraviolet radiation and smoking; injury and disease, medications and other medical treatments such as radiation; strenuous exercise; and physiological stage or conditions such as those in premature babies and the elderly. It is proposed that, in addition to current focus on tissues, the antioxidant status of digesta should be considered because of its effect on specific tissues and potential health implications.

Relative bioavailability of RRR- and all-rac-alpha-tocopheryl acetate in humans: studies using deuterated compounds.

Vitamin E in nutritional supplements in its most common form is alpha-tocopheryl acetate. Available stereoisomeric forms are RRR- (1 stereoisomer) and all-rac- (8 stereoisomers). We evaluated the relative bioavailability of RRR- and all-rac-alpha-tocopheryl acetate using the deuterium-labeled isotopes [5-CD3] 2R, 4'R and 8'R-alpha-tocopheryl acetate (d3), and [5,7-(CD3)2]-all-rac-alpha-tocopheryl acetate (d6). Six adults (three males, three females), aged 25-59 y, received 150 mg each of d3 and d6 for 11 consecutive days. Blood samples were collected on days -1, 0, 1-11, 13, 14, 20, 25, 30, 60, 74, 88, 102, 122, and 137. Plasma and red blood cell tocopherol were evaluated by using HPLC and gas chromatography/mass spectrometry to distinguish between d3 and d6 tocopherols. Cholesterol, triglycerides, and LDL and HDL cholesterol were measured. Relative bioavailability of d3 when compared with d6 was 2.0 +/- 0.06 when area under the plasma time concentration curve (AUC d3/d6) by trapezoidal rule (P < 0.05) was used. Correcting for lipid yielded the same finding. Unlabeled tocopherol (d0) decreased (P < 0.05) with vitamin E administration. It was concluded that the ratio of bioavailability of RRR-/all-rac-alpha-tocopheryl acetate is significantly greater than the currently accepted ratio of 1.36.

Tissue alpha-tocopherol concentrations following supplementation with various forms of vitamin E in sheep.

Tissue alpha-tocopherol concentrations were determined in 40 lambs following oral supplementation of various forms of vitamin E. Lambs were allocated to 8 dietary groups of 5 animals each and supplemented with or without equimolar amounts (300 mg equivalence) of different vitamin E compounds daily for 60 d as follows: 1) control, no supplemental vitamin E; 2) D-alpha-tocopheryl acetate; 3) DL-alpha-tocopheryl acetate; 4) DL-alpha-tocopheryl nicotinate; 5) D-alpha-tocopheryl polyethylene glycol 1,000 succinate (TPGS); 6) DL-alpha-tocopheryl nicotinate+TPGS; 7) D-alpha-tocopheryl acetate+TPGS; and 8) D-alpha-tocopheryl succinate. At the end of the 60 d the lambs were killed and portions of adrenal, fat, heart, kidney, liver, lung, skeletal (brachiocephalicus and gluteus) muscles, pancreas and spleen were removed. Daily supplementation with various vitamin E compounds (equivalence) in lambs resulted in significant differences in tissue alpha-tocopherol concentration in heart, liver, gluteus medius muscle, and spleen. Correlations between the plasma and tissue alpha-tocopherol levels were highly significant for all tissues except adrenal, fat, and pancreas.

Serum total cholesterol, high-density lipoprotein-cholesterol and triglyceride concentrations in lambs following supplementation with various forms of tocopherol.

A 61-d study involving 40 crossbred lambs evaluated the effect of various forms of tocopherol provided daily in equimolar amounts on total cholesterol, high-density lipoprotein-cholesterol and triglyceride concentrations in the serum of lambs. Thirty-five lambs were allotted to 7 treatment groups of 5 animals each, supplemented with 300 mg tocopherol either as: 1) DL-alpha-tocopheryl acetate; 2) D-alpha-tocopheryl acetate; 3) D-alpha-tocopheryl succinate; 4) D-alpha-tocopheryl polyethylene glycol 1,000 succinate (TPGS); 5) DL-alpha-tocopheryl nicotinate; 6) DL-alpha-tocopheryl nicotinate (150 mg) + 150 mg TPGS; and 7-D-alpha-tocopheryl acetate (150 mg) + 150 mg TPGS mixed with the commercial flock diet. In addition, another group of 5 lambs were used as control (no vitamin E supplementation). Dietary supplementation of various vitamin E sources resulted in no overall treatment effects for total cholesterol, triglycerides or high density lipoprotein-cholesterol. A significant variation was noticed among animals. The levels of all measured serum components varied throughout the experimental period (P < 0.0001). The day x treatment interaction was not significant (P > 0.05) for any serum measured component. The present data strongly suggest that short-term treatment (< 2 mo) with pharmacological oral doses of various forms of vitamin E did not influence serum lipid metabolism of lambs. The data also showed that the bioavailability of alpha-tocopherol is dependent on the form administered. D-alpha-tocopherol acetate is a highly available form, the bioavailability of which is further increased when combined with D-alpha-tocopheryl polyethylene glycol succinate.

Cattle blood plasma and red blood cell alpha-tocopherol levels in response to different chemical forms and routes of administration of vitamin E.

The chemical form and site of administration of vitamin E were assessed in cattle by measuring area under the time curve alpha-tocopherol levels in plasma and red blood cells (RBC). In Exp. 1, four groups of Holstein calves, 4 to 6 d old, received daily one of the following forms equivalent to 228 mg of alpha-tocopherol: D-alpha-tocopheryl acetate (DTA); DL-alpha-tocopheryl acetate (DLTA); D-alpha-tocopheryl polyethylene glycol 1,000 succinate (TPGS); and an experimental blend (EB) containing DTA and TPGS. A control group received no supplement. Temporal alpha-tocopherol responses were determined in blood samples collected for 35 d. Relative to controls whose plasma and RBC alpha-tocopherol levels remained unchanged, all supplementation increased (P less than .05) alpha-tocopherol levels in plasma and RBC in calves; the highest levels occurred in those that received either DTA or EB. In a second experiment, five Hereford steers with ruminal and duodenal cannulas were used in a crossover design to determine whether site of vitamin E administration affected plasma alpha-tocopherol levels. A single dose of 810 mg of DTA was placed daily into the rumen, and the same amount divided into five equal doses was given (into the duodenum) during a 12-h period. Plasma alpha-tocopherol levels were higher (P less than .05) when steers were ruminally dosed than when they were dosed duodenally. The results suggest that the alpha-tocopherol concentration of plasma and red blood cells in cattle depends on both the chemical form and on the site of administration.

Bioavailability of vitamin E compounds in lambs.

A study was carried out to assess the bioavailabilities of several forms of vitamin E in lambs. A total of 40 lambs was allotted to eight dietary groups of five each and supplemented or not daily for 60 d with equimolar amounts of different vitamin E compounds as follows: 1) control, no supplemental vitamin E, 2) DL-alpha-tocopheryl acetate, 3) D-alpha-tocopheryl acetate, 4), D-alpha-tocopheryl succinate, 5) D-alpha-tocopheryl polyethylene glycol 1,000 succinate (TPGS), 6) DL-alpha-tocopheryl nicotinate, 7) DL-alpha-tocopheryl nicotinate+ TPGS, or 8) D-alpha-tocopheryl acetate + TPGS. During these 60 d, serum alpha-tocopherol concentrations in the control lambs remained constant and lower (P less than .05) than in lambs that received all treatments. Various indices of bioavailability, including Cmax-C(i) (concentration maximum-concentration initial), Ct-C(i) (concentration terminal-concentration initial), areas under the serum concentrations profiles, and pooled increment were higher (P less than .05) with D-alpha-tocopheryl acetate+ TPGS than in the other groups, suggesting a synergism between these forms. No such effect was observed between nicotinate and TPGS. For the TPGS, a water-soluble form of vitamin E, the indices of bioavailability were lower (P less than .05) than for the other groups. D-alpha-tocopheryl acetate resulted in a bioavailability that outranked all the other forms of vitamin E, except those of D-alpha-tocopheryl acetate + TPGS. A slightly higher bioavailability index was observed for D-alpha-tocopheryl succinate than for DL-alpha-tocopheryl nicotinate.(ABSTRACT TRUNCATED AT 250 WORDS)

Impaired ability of patients with familial isolated vitamin E deficiency to incorporate alpha-tocopherol into lipoproteins secreted by the liver.

Plasma and lipoprotein alpha-tocopherol concentrations of four patients with familial isolated vitamin E deficiency and six control subjects were observed for 4 d after an oral dose (approximately 15 mg) of RRR-alpha-tocopheryl acetate labeled with six deuterium atoms (d6-tocopherol). Chylomicron d6-tocopherol concentrations were similar in the two groups. d6-Tocopherol concentrations of plasma, very low (VLDL), low (LDL), and high (HDL) density lipoproteins were similar in the two groups only during the first 12 h; then these were significantly lower, and the rate of disappearance faster, in the patients. The times (tmax) of the maximum chylomicron d6-tocopherol concentrations were similar for the two groups, but tmax values in the controls increased in the order: chylomicrons less than VLDL less than or equal to LDL approximately HDL, while the corresponding values in the patients were similar to the chylomicron tmax. Thus, plasma d6-tocopherol in controls increased during chylomicron and VLDL catabolism, whereas in patients it increased only during chylomicron catabolism, thereby resulting in a premature and faster decline in the plasma tocopherol concentration due to a lack of d6-tocopherol secretion from the liver. We suggest that these patients are lacking or have a defective liver "tocopherol binding protein" that incorporates alpha-tocopherol into nascent VLDL.

Production responses of dairy cows fed various amounts of rumen-protected methionine and lysine.

A 3 x 3 factorial response surface design was used to study the effects of feeding rumen-protected methionine and lysine to dairy cows between 22 and 305 d of lactation. A total of 130 dairy cows at three universities were individually fed a corn silage and corn grain-based diet that contained either soybean meal or corn gluten meal and urea. An unsupplemented control diet plus nine treatment combinations of three amounts of rumen-protected DL-methionine (3.4, 7.8, and 12.2 g/d) and three amounts of rumen-protected L-lysine (5.9, 13.5, and 21.1 g/d) were fed at all locations. Plasma concentrations of methionine and lysine were increased when rumen-protected methionine and lysine were supplemented to the diets. Rumen-protected methionine and lysine did not affect feed intake by cows fed either by soybean meal or corn gluten meal and urea based diets. Milk protein percentage was increased, but milk and milk protein yields were not improved when diets containing soybean meal were supplemented with rumen-protected methionine and lysine. In contrast, milk and milk protein yields were improved when a diet that contained corn gluten meal and urea was supplemented with rumen-protected methionine and lysine. Health and reproduction measurements were similar for cows receiving all treatments.

Supplementation of dairy cow diets with ammonium salts of volatile fatty acids.

The objective was to determine the efficacy of a blend of ammonium salts of the volatile fatty acids, isobutyric, 2-methylbutyric, isovaleric, and valeric as a supplement to diets for dairy cows. Treatments of 0 (control) or 120 (supplemented) g/cow of the blend were fed daily from approximately 3 wk prepartum through a complete lactation. Five trials were conducted concurrently with a total of 116 multiparous Holstein cows. Dietary ingredients or combinations of ingredients differed in each of the trials. Diets contained either 1) corn gluten meal and urea, 2) soybean meal, or 3) cottonseed meal as the primary grain source of crude protein. The forage portion of the diets contained corn silage in combination with one or more of the following: alfalfa hay, alfalfa haylage, or wheat silage. Cows fed the supplement produced more milk and fat-corrected milk than the control cows for the 305-d lactation on four of the five diets, resulting in an average increase of 1.7 kg/d or 7%. Feed intake of cows on the supplemented diet was generally similar or lower than intake of the control cows throughout lactation, indicating that increased milk yield was associated with improved feed utilization. Percent milk fat was similar for cows on the supplemented diet, but fat yield was higher. Percent milk protein was lower for supplemented cows, but protein yield was about the same for both treatments because of higher milk yield. Health and reproduction were similar for all cows.

Effect of rumen-protected methionine on plasma free amino acids and production by dairy cows.

The effectiveness of an encapsulated rumen-protected methionine compound (RPMet) for delivering methionine postruminally and its effect on plasma amino acid concentrations, milk production and milk composition were evaluated. Thirty multiparous Holstein cows in early lactation were randomly assigned to five levels of RPMet. RPMet was supplied at 0, 550, 1100, 1650 or 3300 mg/kg of concentrate resulting in an intake of 0, 6.9, 12.9, 19.6 or 40.2 g of supplemental DL-methionine/day for each cow. The basal diet was alfalfa-bromegrass hay and a 15.8% crude protein corn- and soybean meal-based concentrate mixture. The experiment was divided into pretreatment and treatment periods, each consisting of 21 days. Feed intake and milk production were measured daily, milk composition weekly and plasma amino acid concentrations twice daily on days 17 and 20 of the pretreatment period and on days 4, 10 and 21 of the treatment period. RPMet was effective in delivering methionine postruminally, which increased concentrations of methionine in plasma. Dry matter intake, milk production, milk composition and body weight were not significantly affected by RPMet.

Production responses of dairy cows fed diets supplemented with ammonium salts of iso C-4 and C-5 acids.

A three-university study examined effects of ammonium salts of volatile fatty acids on dairy cows for a full lactation cycle. Objectives were a) to determine the most efficacious amount and ratio of two components, ammonium salts of C-5 acids and ammonium isobutyrate, and b) to compare responses of this optimum blend with the control in a response surface design. This design tested six blends of the two components defined by the center point and the five points of an equilateral pentagon from which response surfaces would be computed and a control. Each treatment consisted of 23 cows for a total of 161 multiparous Holstein cows. All cows were fed corn silage for ad libitum consumption, 1.8 kg of urea-mineral-vitamin-premix plus supplement based on corn grain and corn gluten meal according to milk production. Response surfaces, based on milk yield, defined an optimum blend composed of 61 g ammonium salts of C-5 acids plus 28 g ammonium isobutyrate per cow per day equivalent to 120 g aqueous blend (74% solids). The optimum blend was similar to the center point blend. Cows receiving the center point blend peaked higher and produced more milk, 4% fat-corrected milk, milk protein, and total solids than control cows. For the first trimester of lactation, milk yields for control and center point blend were 28.6 and 31.8 kg. The 305-day milk, 4% fat-corrected milk yields, and dry matter intakes (kg) were: control 20.5, 19.7, 17.3; center point blend 23.2, 21.6, 17.5. Increased milk production was associated with improved feed utilization. No effect on health or reproduction was observed.