Ronald L. Prior

Plasma Antioxidant Capacity Changes Following a Meal as a Measure of the Ability of a Food to Alter In Vivo Antioxidant Status

Objective: Determine 1) if consumption of a meal of different fruits or berries increases plasma hydrophilic (H-) or lipophilic (L-) antioxidant capacity (AOC) measured as Oxygen Radical Absorbance Capacity (ORACFL); 2) if including macronutrients in the meal alters postprandial changes in AOC; and 3) if preliminary recommendations can be developed for antioxidant intake. Methods: Changes in plasma AOC following consumption of a single meal of berries/fruits (blueberry, dried plum, dried plum juice, grape, cherry, kiwifruit and strawberry) were studied in 5 clinical trials with 6–10 subjects per experiment. In two studies with blueberry or grape, additional macronutrients (carbohydrate, fat, protein) were included in the control and treatment meals. Blood samples collected before and after the meal were analyzed for AOC. Results: Consumption of dried plums or dried plum juice did not alter either the H- or L-AOC area under the curve (AUC). Consumption of blueberry in 2 studies and of mixed grape powder [12.5 (Study #1), 39.9 (Study #4) and 8.6 (Study #5) mmole Trolox Equivalents (TE) AOC, respectively] increased hydrophilic AOC AUC. L-AOC increased following a meal of blueberry containing 12.5 mmole TE AOC (Study #1). Consumption of 280 g of cherries (4.5 mmol TE AOC) increased plasma L-AOC but not H-AOC. The AOC in the control groups in which additional macronutrients (Studies #4 and #5) were added decreased from the postprandial baseline AOC measurement. Conclusion: We have demonstrated that consumption of certain berries and fruits such as blueberries, mixed grape and kiwifruit, was associated with increased plasma AOC in the postprandial state and consumption of an energy source of macronutrients containing no antioxidants was associated with a decline in plasma AOC. However, without further long term clinical studies, one cannot necessarily translate increased plasma AOC into a potential decreased risk of chronic degenerative disease. Preliminary estimates of antioxidant needs based upon energy intake were developed. Consumption of high antioxidant foods with each meal is recommended in order to prevent periods of postprandial oxidative stress.

Disease incidence and longevity are unaltered by dietary antioxidant supplementation initiated during middle age in C57BL/6 mice

The ability of augmented antioxidant consumption to alter disease incidence, lesion burden and/or longevity was studied in adult male C57BL/6 mice. Mice were fed modified AIN76 diet or modified AIN76 supplemented with vitamin E, glutathione (GSH), vitamin E and GSH, melatonin or strawberry extract starting at 18 months of age. All the mice in this study were heavier than reference populations of male C57BL/6 mice fed NIH-07 or NIH-31, which were maintained without a mid-life change in diet. Fatty liver, focal kidney atrophy and proteinacious casts in the renal tubules were observed more frequently in this study population than in the reference populations. Lesion burden and incidence of specific lesions observed amongst the various groups in this study did not differ. There were no differences observed for longevity of any of the study groups. The longevity observed in this study was similar to that previously reported for male C57BL/6 mice. Thus, diet supplementation with antioxidants initiated during middle age did not appear to affect age-associated lesions patterns, lesion burden or longevity for ad libitum fed male C57BL/6 mice.

AGE-RELATED NEURODEGENERATION AND OXIDATIVE STRESS: Putative Nutritional Intervention

This review describes age-related changes that occur in neuronal function and cites evidence to show that these alterations may be the result of increased sensitivity to oxidative stress (OS). Evidence is presented to show that the abilities to mitigate the OS effects and to repair the damage from OS show decline as a function of age. Results from age- and OS-sensitive tests are given; these results indicate that one of the major sites of action of OS is the membranes, especially if compromised by high amounts of sphingomyelin, and one of the major effects of OS is to further alter the calcium disregulation in aging. It is suggested that attempts to increase antioxidant protection through diets comprised of fruits and vegetables identified as being high in total antioxidant activity might prevent or reverse the deleterious OS effects on neuronal aging.

The effects of a multivitamin/mineral supplement on micronutrient status, antioxidant capacity and cytokine production in healthy older adults consuming a fortified diet.

Background: Inadequate micronutrient intake among older adults is common despite the increased prevalence of fortified/enriched foods in the American diet. Although many older adults take multivitamin supplements in an effort to compensate, studies examining the benefits of this behavior are absent. Objective: To determine whether a daily multivitamin/mineral supplement can improve micronutrient status, plasma antioxidant capacity and cytokine production in healthy, free-living older adults already consuming a fortified diet. Methods: An eight-week double-blind, placebo-controlled clinical trial among 80 adults aged 50 to 87 years (mean=66.5±8.6 years). Results: Multivitamin treatment significantly increased (p<0.01, compared to placebo) plasma concentrations of vitamins D (77 to 100 nmol/L), E (27 to 32 μmol/L), pyridoxal phosphate (55.1 to 75.2 nmol/L), folate (23 to 33 nmol/L), B12 (286 to 326 pmol/L)), C (55 to 71 μmol/L), and improved the riboflavin activity coefficient (1.23 to 1.15), but not vitamins A and thiamin. The multivitamin reduced the prevalence of suboptimal plasma levels of vitamins E (p=0.003), B12 (p=0.004), and C (p=0.08). Neither glutathione peroxidase activity nor antioxidant capacity (ORAC) were affected. No changes were observed in interleukin−2, −6 or −10 and prostaglandin E2, proxy measures of immune responses. Conclusions: Supplementation with a multivitamin formulated at about 100% Daily Value can decrease the prevalence of suboptimal vitamin status in older adults and improve their micronutrient status to levels associated with reduced risk for several chronic diseases.

Effect of vitamin E intake on levels of vitamins E and C in the central nervous system and peripheral tissues : implications for health recommendations

Vitamin E (alpha-gamma-tocopherol) is an important component in biological membranes. A decrease in its concentration imposes structural and functional damage to the cells. The object of this study was to assess the effect of a graded dietary vitamin E (E) intake on E concentration in specific regions of the brain, and its influence on vitamin C levels and neurological function. Following a 2-month period, rats supplemented with 5, 30, 60, 250 or 500 mg all-rac-alpha-tocopherol-acetate/kg diet (mg E/kg diet) exhibited a significant increase of E concentration in brain and peripheral tissues. However, while blood and liver showed a dose response increase in E concentration which correlated well with the different levels of E in the diet, the central nervous system (CNS) followed the same pattern of increase of vitamin E in brain tissue only when the diet was supplemented with 5, 30, or 60 mg E/kg diet. No further increase in E concentration was observed when the diet was supplemented with 250 or 500 mg E/kg diet. Similarly, the heart tissue showed a significant increase in its E concentration when the was enriched with 5, 30, or 60 mg E/kg diet, with no further increases at 250 or 500 mg. Vitamin C concentration in brain cortex and cerebellum, plasma, liver, and heart was reduced in the groups receiving 250 or 500 mg E/kg diet. Compared to the low E group, rats supplemented with the 60, 250 or 500 mg E/kg diet showed a significant enhancement in striatal dopamine (DA) release, but no differences were observed among the latter three groups.

Membrane and Receptor Modifications of Oxidative Stress Vulnerability in Aging: Nutritional Considerations

ABSTRACT: Evidence suggests that oxidative stress (OS) may contribute to the pathogenesis of age‐related decrements in neuronal function and that OS vulnerability increases as a function of age. In addition to decreased endogenous protection, increases in OS vulnerability may result from changes in membrane lipids and distribution of receptor subtype. Using a PC‐12 cell model system, we have shown that H2O2 or dopamine (DA) exposure induced deficits in the cells ability to clear (extrude/sequester, E/S) Ca2+ that are similar to those seen in aging. When plasma membrane concentrations of sphingomyelin (SPM) were used, the SPM metabolite, sphingosine‐1‐phosphate was increased to the same levels as those seen in aging, and enhancement of OS‐induced decreases in calcium E/S following KCL depolarization was observed. Differential decreases in CA2+ E/S were also seen following DA‐induced OS in COS‐7 cells transfected with one of five muscarinic receptor subtypes. Cells transfected with either M1, M2, or M4 receptors showed significantly greater vulnerability to OS (as expressed by greater decrements in calcium E/S and cell death) than those transfected with M3 or M5 receptors. The vitamin E analogue, Trolox, and the nitrone‐trapping agent, PBN, were not effective in altering E/S decrements but were effective in preventing cell death 24 h after OS exposure. These findings suggest that putative regional (e.g., striatum and hippocampus) increases in OS vulnerability and loss of neuronal function in aging may be dependent upon membrane SPM concentration and receptor subtype. In related studies, attempts were made to determine whether increased OS protection via nutritional increases in antioxidant levels in rats [using diets supplemented with vitamin E (500 IU/kg), strawberry extracts (9.4 g/kg dried aqueous extract, DAE), spinach (6.7 g/kg DAE), or blueberry extracts (10 g/kg DEA for six weeks)] would protect against exposure to 100% O2 (a model of accelerated neuronal aging). Results indicated that these diets were effective in preventing OS‐induced decrements in several parameters (e.g., nerve growth factor decreases), suggesting that although there may be increases in OS vulnerability in aging, phytochemicals present in antioxidant‐rich foods may be beneficial in reducing or retarding the functional central nervous system deficits seen in aging or oxidative insult.

Antioxidant capacity in different tissues of young and old rats.

Abstract The antioxidant capacity in heart, liver, lung, and kidney was studied in young (6 months) and old (22 months) male Fischer 344 rats, using the oxygen radical absorbance capacity (ORAC) assay system with two different reactive oxygen species (ROS) generators. The results indicated that liver in old rats had significantly lower peroxyl radical absorbance capacity (ORACROO, units/g wet wt), but higher hydroxyl radical absorbance capacity (ORACOH, units/mg protein) than in young rats. The decreased liver ORACROO in the old rats was mainly due to the loss of cytosol protein, while the increased liver ORACOH in the old rats was a result of an increased resistance of cytosol proteins to the attack of ROS. This conclusion was further supported by the finding that the contribution of nonprotein fraction of liver cytosol to the ORACOH of the cytosol decreased with age. No effect of age was found on either ORACROO or ORACOH in other tissues. The antioxidant capacity for both ORACROO and ORACOH, was usually high in liver and kidney but low in lung and heart. [P.S.E.B.M. 1996, Vol 211]

ANTIOXIDANT CAPACITY DECREASES DURING GROWTH BUT NOT AGING IN RAT SERUM AND BRAIN

The objective of this study was to evaluate the effects of growth and aging on the antioxidant capacities of rat serum and brain, by using two different reactive oxygen species (ROS) generators and two different methods of expressing the results. The antioxidant capacity was determined by using the oxygen radical absorbance capacity (ORAL) assay. The protein content of serum increased significantly during the growth period of day 1 to 6 months of age. The protein content in brain (cortex and cerebellum) extract also tended to increase with the growth. The antioxidant capacity of serum decreased during growth, if the ORAC values were expressed on the basis of serum protein concentration. The antioxidant capacity in brain cortex and cerebellum declined significantly during growth, regardless of which ROS generator was used in the ORAC assay or how the results were expressed (units/ml or units/mg protein). Therefore, the rat pups had higher antioxidant capacity than the adult and old rats. The low plasma antioxidant capacity reported in premature neonates may simply result from a low protein concentration of the plasma, which occurs in neonates. No aging effect was observed on the antioxidant capacity of serum and brain in the adult and old rats.

Responses of circulating urea cycle and branched-chain amino acids to feeding in adult and aged Fischer-344 rats

Amino acid concentrations in plasma and blood were compared between adult (6 months) and aged (22 months) male rats in fed and fooddeprived (fasted) states. Since feeding is known to elevate plasma amino acids, rats were sampled after food had been freely available to them and after food was withheld for 20 hours. Fourteen amino acids increased (8 to 74%) in blood and plasma in fed rats compared to food-deprived rats. Blood concentrations of four amino acids (cystine, aspartate, threonine, and phosphoserine) were increased in aged compared to adult rats, while tyrosine was 10% lower. The interaction between fed state and age was significant for the branched-chain amino acids (BCAA) as they increased 23% in the blood and plasma of fed compared to food-deprived adult rats; however, BCAA were not affected by feeding state in the aged rats. Plasma arginine concentration was 12% lower (p<0.001) and citrulline was 20% higher (p<0.005) in aged compared to adult rats, and consequently the citrulline:arginine ratio was elevated by 36% in aged rats. Our results demonstrate that, although the total arginine in whole blood is not altered or is increased with aging, less of the arginine is transported in the plasma of aged compared to adult rats. Furthermore, these results indicate altered metabolism of the urea cycle amino acids and BCAA with aging in the rat.