Evaluation of palatability and muscle composition of novel value-added beef cuts.

The aim of this study was to evaluate the muscle profile of novel added-value beef cuts including the caudal tip of the M. infraspinatus (Bonanza Cut; TIP) and M. subscapularis (SUB) and two traditional sirloin steak cuts, M. gluteus medius (top sirloin; GLM) and M. rectus femoris (sirloin tip; REC). Samples were subjected to Warner-Braztler Shear Force (WBSF), sensory, cooking loss, and proximate analysis. The muscle TIP had superior values of subjective tenderness, juiciness, and slight off-flavor intensity when compared to all other muscles. The TIP and SUB were similar in WBSF. Cooking loss and moisture values of raw samples were lowest for TIP. Results suggest that TIP can provide enhanced eating experience for consumers and improved marketability for the meat industry.

Urinary hydroxyproline excretion and vitamin C status in healthy young men.

The relationship between ascorbic acid status and the urinary excretion of hydroxyproline was examined in 11 healthy male subjects fed an ascorbic acid-deficient diet for 14 wk while in a metabolic unit. The diet provided 5 mg ascorbic acid/d and was supplemented with ascorbic acid to give intakes of 65 mg/d (2 wk), 5 mg/d (4 wk), 605 mg/d (3 wk), 5 mg/d (4 wk), and an average 375 mg/d (1 wk). The urinary excretion of hydroxyproline increased by an average of 16% and 30% after the first and second depletion periods, respectively, and decreased to baseline values after supplementation with normal or high doses of vitamin C. Significant (p less than 0.05) inverse correlations were found between urinary hydroxyproline and plasma, red cell, and leukocyte ascorbic acid. These results show that urinary hydroxyproline excretion increases during human vitamin C deficiency but that this effect is not strong enough to provide a reliable marker of mild vitamin C deficiency.

Biochemical indices of human vitamin C status.

Biochemical indicators of ascorbic acid (AA) status were studied in eleven young adult males fed the same AA deficient diet for 14 wk in a live-in metabolic unit. Supplements of AA were added to the diet to give AA-intake periods of 65 mg/d (2 wk), 5 mg/d (4 wk), 605 mg/d (3 wk), 5 mg/d (4 wk), 605 mg/d (4 d), and 65 mg/d (3 d). Blood plasma, erythrocyte, and leukocyte AA levels all reflected AA intake, however, plasma AA showed less variability than red cell AA levels and was considerably easier to determine than leukocyte AA. Plasma AA values less than 0.40 mg/dL (23 mumol/L) reflected marginal AA status. The daily AA intake calculated to maintain plasma AA levels of at least 0.4 mg/dL (23 mumol/L) in healthy young men was 41 mg. The average AA intake estimated to maximize the total body pool was 138 mg/d. Urine and salivary AA levels were not useful indicators of AA status because urinary AA levels did not discriminate well between adequate and deficient AA intakes and salivary AA levels did not consistently reflect AA intake.

Plasma ascorbic acid in adult males: effects of depletion and supplementation.

We investigated effects of moderately elevated oral doses of ascorbic acid (AS) on plasma AS turnover in healthy men after abrupt alterations in AS intake. Subjects ate a basal diet of conventional food in which only AS intake was changed. Blood specimens were collected from fasted subjects twice each week during depletion periods. Deproteinized plasma was analyzed after derivatization with 2,4-dinitrophenylhydrazine. Subjects remained healthy and never became frankly scorbutic. The kinetics of log-converted plasma AS values for the depletion periods demonstrated that the slopes calculated by least squares were up to 56% higher for six of nine subjects after 600 mg AS/day. The difference between the mean slopes (M) for loss of plasma AS was significantly different (p less than 0.05, n = 6) after 60 mg AS (M = -0.0222 +/- 0.0145) than after 600 mg AS (M = -0.0246 +/- 0.0003). Leukocyte AS concentration decreased 44.6% in the first depletion period and 77.1% in the second.

Effect of ascorbic acid on copper and cholesterol in adult cynomolgus monkeys fed a diet marginal in copper.

The effects of prolonged consumption of high levels of dietary ascorbic acid on copper metabolism and cholesterol in adult monkeys fed a diet low or marginal in copper were investigated. Small reductions in serum copper and in serum ceruloplasmin levels were observed when high levels of ascorbic acid were fed. During the period of copper depletion there was a gradual but significant (p less than 0.001) increase in serum cholesterol. The level of ascorbic acid supplementation had no effect during this phase. When copper was added back to the diet, serum cholesterol levels leveled off or declined in the monkeys receiving the low (1 mg/day/kg body weight) dose of ascorbic acid. Cholesterol levels continued to increase in the group receiving the higher ascorbic acid supplement (25 mg/day/kg body weight). These data suggest that high levels of ascorbic acid supplementation may make dietary copper relatively unavailable for regulating cholesterol metabolism.

Measurement of vitamin C in blood components by high-performance liquid chromatography. Implication in assessing vitamin C status.

The fact that platelets, PMN leukocytes, and MN leukocytes concentrate ascorbic acid suggests that vitamin C has an important role in their physiological functions. The question still remains as to which one of the cells best reflects vitamin C status. The ascorbic acid content of PMNs and platelets correlates positively with plasma concentration and supplementation with vitamin C, as shown in Evans et al. They also found that MN leukocytes, in contrast, do not show any such relationship; however, MN leukocytes maintain the highest levels of ascorbic acid and play a very important function in immunocompetence. We have found that with a limited number of subjects, ascorbic acid content of MN and PMN leukocytes correlates positively with plasma ascorbic acid, but there was no correlation between platelets and plasma ascorbic acid (unpublished results). Therefore, further work is necessary to evaluate these three blood components for the best cellular marker of vitamin C status. We have developed a reversed-phase HPLC method for ascorbic acid that can be used in conjunction with our cellular differential centrifugation technique for the determination of ascorbic acid in relatively pure blood cell fractions. The chromatographic method is simple, sensitive, and automated. It clearly resolves ascorbic acid, which is the major form of the vitamin found in vivo and is not prone to interference by sugars, carbohydrates, or nucleotides.

Experimental vitamin C depletion and supplementation in young men. Nutrient interactions and dental health effects.

Biochemical indices of AA clearly showed that the young men in this study were brought into various states of AA depletion and repletion according to their dietary AA intakes. While previous studies have postulated that supplemental intakes of AA may adversely affect body status of vitamins B6 and B12, we found no changes in the B vitamin status of the young men receiving varying AA intakes. Moderate AA supplementation (605 mg/day) showed no antagonistic effect on markers of vitamins B6 and B12. Blood markers of fat-soluble vitamins A and E and iron status were not affected by AA intakes. The propensity of the gingiva to become inflamed or bleed on probing was reduced after normal (65 mg/day) AA intakes as compared to deficient (5 mg/day) intakes and upon supplementary (605 mg/day) AA intakes as compared to normal intakes. The results suggest that AA status may influence early stages of gingival inflammation and crevicular bleeding, and warrant further study of the relationship between AA and periodontal health.

Beta-carotene and protein oxidation: effects of ascorbic acid and alpha-tocopherol.

The effect of beta-carotene on protein oxidation was examined under different oxygen (O(2)) tensions and with other antioxidants: alpha-tocopherol, ascorbic acid, and mixtures of antioxidants. Human serum albumin (HSA) was incubated with 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) to induce protein oxidation (carbonyl formation), under 15, 150, and 760 torr of O(2) tension. Antioxidant activity was related to O(2) tension, antioxidant concentrations and interaction between mixtures of antioxidants: (1) Under 15 torr of O(2), incubating HSA with AAPH, 1. 6 microM beta-carotene, 80 microM alpha-tocopherol, 160 microM ascorbic acid, and mixtures (0.1 microM beta-carotene, 5.0 microM alpha-tocopherol and 10 microM ascorbic acid) resulted in 24, 29, 39, and 44% reduction of carbonyl formation, respectively. (2) Under 150 torr of O(2) tension, the antioxidant effect of beta-carotene was decreased by 4% but increasing O(2) tension did not diminish the antioxidant effects of alpha-tocopherol, ascorbic acid, or antioxidant mixtures. (3). Under 760 torr of O(2) tension, adding 1. 6 microM beta-carotene resulted in 26% more carbonyl formation. (4) Under 760 torr of O(2) tension, the antioxidant effect of ascorbic acid was decreased 32% compared to what was observed at 150 torr of O(2) tension. Changes in O(2) tension had no effect on the antioxidant effect of alpha-tocopherol. The mixture of antioxidants inhibited carbonyl formation by 37% and was 7% less effective than that of 15 and 150 torr of O(2) tension. High concentration of beta-carotene produces more protein oxidation in the presence of high O(2) tension by a prooxidant mechanism. Mixtures of beta-carotene, alpha-tocopherol, and ascorbic acid provided better protective effects on protein oxidation than any single compound.

Free radical-mediated lung response to the monofunctional sulfur mustard butyl 2-chloroethyl sulfide after subcutaneous injection.

Vesicant-induced pathogenesis is initiated by rapid alkylation and cross-linking of DNA purine bases causing strand breaks leading subsequently to NAD depletion and cell death. We postulated that vesicants may also be associated with free radical-mediated oxidative stress distal to the site of exposure. To test this postulate in the lung, we injected 3 groups (n = 8) of 5-month-old, male, athymic, nude mice, weighing 30-35 g with a single subcutaneous (s.c.) injection (5 microliters/mouse) of butyl 2-chloroethyl sulfide (BCS), a monofunctional sulfur mustard analog. After 1, 24 and 48 h, we euthanized the treated mice along with 2 untreated control mice at each time point. We then pooled the control mice in one group (n = 6) and analyzed the lungs for biochemical indices of oxidative stress. We found that total lung weight was not altered after treatment, but wet/dry weight ratio decreased 18% (P less than 0.05) and hemoglobin content increased 50% and 36% at 1 and 24 h, respectively. The activity of glucose-6-phosphate dehydrogenase increased significantly, 40% at 1 and 24 h and 84% at 48 h and that of glutathione S-transferases was 60%, P less than 0.05 greater at all time points. Lipid peroxidation (estimated by the thiobarbituric acid test) and total protein content increased 3-fold and 2-fold, at 1 and 24 h, respectively. Total and oxidized glutathione contents were significantly elevated, 38% at 1 h and 64% at 24 h for the former and 45% at 24 h and 56% at 48 h for the latter. Because these changes are consistent with the cellular response to oxidative stress, we conclude that BCS injected subcutaneously, can cause changes in the lung possibly via a free radical-mediated mechanism.

Kinetics of selenium incorporation into tissues of female mallard ducks.

Selenium is essential for both mammalian and avian species, although its metabolism in birds has been less thoroughly studied. Little information has been available on the kinetics of selenium in birds, especially as it relates to the teratogenicity seen in waterfowl consuming excessive amounts. This study describes the pharmacokinetics of small amounts of 75Se as selenious acid injected into female mallard ducks. Labeled selenium was injected into a wing vein of restrained animals and tissues taken at five different time points up to 24 h post-injection. Selenium levels as percent of injected dose were determined in liver, kidney, heart, lung, adrenals, thyroid, spleen, pancreas, ovaries, intestine, muscle and plasma. Estimates of kinetic parameters (uptake and elimination rates, time of maximum concentration and maximum concentration) were obtained with a non-linear kinetics computer program (PCNONLIN, SCI Software, Lexington, KY). Results showed four basic patterns of distribution, uptake and elimination. Visceral tissues exhibited a triphasic pattern with a rapid rise, a decline followed by a distinctive increase in levels between the last two time points. Brain uptake was continuous over the 24 h. Plasma rose rapidly and then declined to a constant level. The ovaries as a tissue of interest relating to the teratogenic effects of selenium showed the greatest relative increase at 24 h, suggesting kinetic mechanisms consistent with a pathway that might lead to accumulation of toxic levels and teratogenic effects during embryo development.

Response of mouse brain to a single subcutaneous injection of the monofunctional sulfur mustard, butyl 2-chloroethyl sulfide (BCS)*.

Exposure to mustard-type vesicants results in alkylation of DNA and vesication. However, the biochemical mechanism for vesicant injury and whether it is localized or diffuse are not clear. We postulated that vesicant damage is mediated by free radicals, resulting in oxidative stress. These free radicals-mediated reactions may propagate systemically distal to the site of exposure. To test this hypothesis, we examined the effects of a single subcutaneous injection of the monofunctional sulfur mustard, butyl 2-chloroethyl sulfide (BCS), on the brain. We injected 3 groups (6 mice/group) of 5-month-old male, athymic, nude mice, weighing 30-35 g, subcutaneously with neat (undiluted) BCS (5 microliters/mouse). After 1, 24, and 48 h, we sacrificed the treated mice along with an untreated control group and analyzed the brains for biochemical markers of oxidative stress. Compared to untreated controls, the activity of glutathione peroxidase increased by 76%, P less than 0.005 at 24 h, and that of glutathione S-transferases by 25-37%, P less than 0.05 over the entire period. Total glutathione content in the brain was significantly lower, 17%, after 1 h and 23% after 24 h. We found also, concomitant with decreased glutathione, almost a 3-fold increase in susceptibility to lipid peroxidation. Because these changes are consistent with oxidative stress, we conclude that the effect of BCS administered subcutaneously may be translocated, reaching mouse brain, and causing oxidative stress.

Antioxidant and prooxidant roles for beta-carotene, alpha-tocopherol and ascorbic acid in human lung cells.

Experiments were conducted to determine the antioxidant and prooxidant effects of beta-carotene, alpha-tocopherol and ascorbic acid on human lung cells at different oxygen (O(2)) tensions. Free radical initiator, 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH), was used to induce the cellular damage associated with lipid peroxidation, protein oxidation and DNA breaks. Under hypoxic conditions (0 torr O(2) tension) all compounds produced a concentration-dependent antioxidant effect. Mixtures of the three compounds exhibited greater protective affects than any individual compound. At 143 torr O(2) tension, all compounds exhibited concentration-dependent protective effects against AAPH-induced cellular lipid, protein and DNA damage. At 722 torr O(2) tension, cells exhibited a consistent increase in lipid peroxidation (isoprostane formation), protein oxidation (carbonyl formation) and DNA damage (p53 protein accumulation). beta-Carotene (1.5 microM) produced a prooxidant effect by promoting 12% isoprostane formation. Protein oxidation and DNA damage at 722 torr O(2) tension was not increased by beta-carotene; however, the antioxidant effect of beta-carotene was attenuated. The antioxidant effects of alpha-tocopherol, ascorbic acid, and mixtures of the three antioxidant compounds also were reduced by the high O(2) conditions. These results partially substantiate the hypothesis that the antioxidant and prooxidant effects of beta-carotene are dependent on O(2) tension and concentration of beta-carotene. Such findings may partially explain why selected populations, such as smokers, respond adversely when supplemented with beta-carotene.

DNA strand breakage and oxygen tension: effects of beta-carotene, alpha-tocopherol and ascorbic acid.

DNA damage is involved in carcinogenesis, aging and other degenerative diseases. The relationship between DNA strand breakage and beta-carotene (0.1-1.6 microM) was examined under different O(2) tensions and with other antioxidants: alpha-tocopherol (5-80 microM), ascorbic acid (10-160 microM) and mixtures of these antioxidants. Supercoiled plasmid DNA pBR322 was incubated with 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) to induce DNA strand breaks in the presence of antioxidants under 15, 150, and 760 torr of O(2) tension. Under 15 torr of O(2) tension, beta-carotene, alpha-tocopherol, ascorbic acid and mixtures of these antioxidants provided a dose-dependent protection against AAPH-induced DNA strand breaks. The best protection was achieved in the mixture of antioxidants. Under 150 torr of oxygen tension, the antioxidant effect of beta-carotene was diminished at > or = 0.8 microM. A prooxidant effect was found at 0.8 > or = microM beta-carotene, producing more single- and double-strand breaks. alpha-Tocopherol and ascorbic acid exhibited dose-dependent antioxidant effects at 150 torr of oxygen tension. Under 760 torr of O(2) tension, the prooxidant effect of 0.8 microM beta-carotene was significant, causing supercoiled DNA to completely breakdown to circular and linear forms. In addition, 760 torr of O(2) tension attenuated the antioxidant effects of alpha-tocopherol and ascorbic acid. Thus, beta-carotene causes concentration-dependent DNA breakdown at high O(2) tension. The protection of DNA from the prooxidant effects of beta-carotene afforded by alpha-tocopherol and/or ascorbic acid was limited at high O(2) tension.

Comparative pharmacokinetics of vitamin A (retinol) in rats and hamsters.

The absorption and elimination of all-trans-retinol in the plasma of rats and hamsters were studied after an oral dose of 45 mg/kg body weight. The hamsters exhibited a higher pretreatment mean circulating retinol concentration than rats maintained on an identical diet. The increase in plasma retinol after a single oral dose was much greater in hamsters than rats. The area under the plasma concentration v. time curve was approximately 60% greater for rats than for hamsters. The elimination half-times for rats were much longer than for hamsters. Plasma retinol uptake and disappearance were consistent with a two-compartment open model with first-order absorption.

Absorption, distribution and elimination of selenium as L-selenomethionine in non-human primates.

20 adult female macaques (Macaca fascicularis) were given oral doses of L-selenomethionine (L-SeMet) equivalent to 0, 25, 150, 300 and 600 micrograms selenium (Se)/kg body weight, and plasma, erythrocyte, hair, faecal and urine Se concentrations were determined. The macaques were scheduled for 30 daily oral doses of L-SeMet, but systemic toxicity necessitated dose reduction in several animals; two macaques given 600 micrograms Se/kg body weight/day for 10-15 days died, and the concentration of Se in their tissues was determined and compared with Se concentrations in tissues collected from one untreated animal. Circulating and urinary Se concentrations in control macaques were within the normal human ranges. Plasma, erythrocyte, hair and urinary Se concentrations were generally dependent on the dose of L-SeMet administered. Plasma Se reflected more immediately exposure to L-SeMet, whereas erythrocyte Se concentrations increased and decreased more slowly. In some cases, erythrocyte Se was still increasing or showed a plateau after L-SeMet treatment was discontinued. Plasma Se concentrations of 6.7-7.3 ppm were observed in the two animals that died due to acute toxicity to L-SeMet. Neither plasma nor erythrocyte GPx activity was influenced by a single L-SeMet dose, but an increase in erythrocyte GPx activity occurred with continuous exposure. Total tissue Se increased 13-28-fold in macaques given 600 micrograms Se/kg body weight/day for 10-15 days, with the liver and kidneys containing the the highest Se concentrations.

Cardiovascular disease hospitalization and ambient levels of carbon monoxide.

Recent research suggests that some cases of cardiovascular mortality may be related to carbon monoxide (CO) air pollution. Clinically based studies indicate the adverse effects of CO on the cardiopulmonary system. However, little attention has been paid to the question of hospital admissions for cardiovascular illness caused by ambient CO levels. The present study assesses the association between hospital admissions for cardiovascular system illnesses and the ambient levels of CO in the Reno-Sparks, NV, area over a 6-yr period (1989-1994). Daily admissions to all three hospitals in the region and daily ambient concentrations of CO, monitored at five sites, were included. There were 32,705 total cardiovascular (CV) admissions, including 13,108 with the diagnosis of ischemic heart disease (IHD) during the study period. The average daily 1-h maximum level of CO was 3.09 ppm. After adjusting for day-of-the-week and seasonal effects and controlling for the effects of autocorrelation errors, both weighted least squares (WLS) and autoregressive integrated moving average (ARIMA) methods showed consistently positive relationships between the ambient CO level and different groups of cardiovascular admissions, although the male gender and age older than 60 groups tended to be most affected. Data suggest a positive correlation between ambient CO levels and hospital admissions for CV diseases.

Distribution of vitamins A and E in blood and liver of rats depleted of vitamin A or vitamin E.

Young Sprague-Dawley male rats (n = 150) were fed a semipurified diet, either without vitamin A (VA), without vitamin E (VE) or supplemented with both vitamins A and E (control). At the end of weeks 0, 1, 3, 5, 6, 7, 8 and 9, groups of rats were anesthetized with methoxyflurane, and blood was collected by cardiac puncture until the rat was exsanguinated. The liver was excised. Whole blood (WB) from each rat was fractionated into plasma (PLA), leukocytes (LEU), platelets (PLT) and erythrocytes (RBC). Each blood component was extracted with heptane and livers were extracted with CHCl3/CH3OH (2:1, v/v). The extracts were analyzed for VA and VE by high performance liquid chromatography. The relationship among blood components in the loss of VA was PLT = LEU greater than WB greater than PLA. The relationship among blood components in the loss of VE was PLA greater than RBC greater than WB greater than LEU = PLT. VA and VE levels in other blood components decreased precipitously between weeks 0 and 4 in the animals placed on deficient diets. These results and correlation analyses between vitamin contents of blood components and of livers indicate inadequacies for the use of certain blood components as monitors of lipid-soluble vitamin status in the rat.

Use of antioxidants in the analysis of vitamins A and E in mammalian plasma by high performance liquid chromatography.

A simple, sensitive, quantitative method for the simultaneous assay of retinol, alpha-tocopherol and gamma-tocopherol in rat, guinea pig, monkey and human plasma was developed by using high performance liquid chromatography. It was found that antioxidant was required to stabilize the fat-soluble vitamins in the plasma of rats. The effect of several antioxidants on the recovery of fat-soluble vitamins was evaluated. Results showed that 0.125% butylated hydroxytoluene (BHT) in ethanol and 0.025% BHT in heptane yielded recoveries greater than 95% in 0.1 ml plasma.

Air pollution and health effects in northern Nevada.

The issue of adverse health effects of ambient air pollution has been extensively studied and reported worldwide over the past two decades. The urban area of Reno and Sparks in northern Nevada is one of two major urban centers in Nevada; the other is Las Vegas. The northern area, which has undergone a rapid population growth in the last decade, has special geographic characteristics and air pollution patterns. We conducted environmental epidemiological studies spanning the 1990s. This report summarizes the evidence and discusses the findings in relation to other studies. Ambient air pollution levels, even when below federal standards, have a marked potential to impact human health adversely. Air pollution was associated with (1) emergency room visits for asthma; (2) hospitalization for chronic obstructive pulmonary disease; (3) hospitalization for cardio-vascular disease; (4) elementary school absenteeism; and (5) low birth weight, preterm birth, and other adverse birth outcomes.

Safety of megavitamin therapy.

Carbon compounds that are needed in small amounts in the diet because they are not made in the body of vertebrates are defined as vitamins. Excluded from this definition are vitamins D, K, and niacin which can be synthesized by the organism or, as in the case of vitamin K, by the host's intestinal bacteria. Lack of such vitamins can result in characteristic deficiency diseases. The therapeutic use of such compounds (megavitamin intake) is based on the spectacular effect of vitamins on deficiency diseases; however, evidence that the ingestion of large amounts of vitamins beyond the "Recommended Daily Allowances" (RDA) is beneficial is not within the basic concept of nutrition. Vitamins, like many substances, may be toxic when taken in large quantities, especially the fat-soluble vitamins, and the concept of "more is better" is a common misconception. Vitamin supplements can be suggested only in the unusual cases of patients having inadequate intake, disturbed absorption (genetic or otherwise), or increased tissue requirements. A well-balanced diet that includes a wide variety of foods from each of the four food groups is adequate for the supply of vitamins, as well as other nutrients, in healthy people. This paper will review some of the recent findings regarding vitamin toxicity and the mechanisms of toxicity.