α-Toc Levels Research Articles

Foliar Spray of Alpha-Tocopherol Modulates Antioxidant Potential of Okra Fruit under Salt Stress.

As an antioxidant, alpha-tocopherol (α-Toc) protects plants from salinity-induced oxidative bursts. This study was conducted twice to determine the effect of α-Toc as a foliar spray (at 0 (no spray), 100, 200, and 300 mg L−1) to improve the yield and biochemical constituents of fresh green capsules of okra (Abelmoschus esculentus L. Moench) under salt stress (0 and 100 mM). Salt stress significantly reduced K+ and Ca2+ ion concentration and yield, whereas it increased H2O2, malondialdehyde (MDA), Na+, glycine betaine (GB), total free proline, total phenolics, and the activities of catalase (CAT), guaiacol peroxidase (GPX), and protease in both okra varieties (Noori and Sabzpari). Foliar application of α-Toc significantly improved the yield in tested okra varieties by increasing the activity of antioxidants (CAT, GPX, SOD, and ascorbic acid), accumulation of GB, and total free proline in fruit tissues under saline and non-saline conditions. Moreover, α-Toc application as a foliar spray alleviated the adverse effects of salt stress by reducing Na+ concentration, MDA, and H2O2 levels and improving the uptake of K+ and Ca2+. Among the tested okra varieties, Noori performed better than Sabzpari across all physio-biochemical attributes. Of all the foliar-applied α-Toc levels, 200 mg L−1 and 300 mg L−1 were more effective in the amelioration of salinity-induced adverse effects in okra. Thus, we concluded that higher levels of α-Toc (200 mg L−1 and 300 mg L−1) combat salinity stress more effectively by boosting the antioxidant potential of okra plants.

Alpha-tocopherol fertigation confers growth physio-biochemical and qualitative yield enhancement in field grown water deficit wheat (Triticum aestivum L.)

Water stress is a major problem to fulfill the world food demand and to solve the problem of malnutrition. Different strategies are being used to solve these problems including the fertigation of plants with different biochemical at different growth stages. The present study was conducted for the induction of drought tolerance in field grown wheat for better yield and nutritional quality through foliar spray of α-tocopherol (α-Toc) at start of reproductive stage. Water stress was maintained based on number of irrigation. Three levels of α-Toc 0.001, 0.01 and 0.1 mM were applied as foliar spray. Water stress significantly reduced the biomass production that associated with the decreased photosynthetic pigments, water relation, photosynthetic efficiency, but increased the lipid peroxidation, leaf relative membrane permeability, activities of antioxidant enzymes and the contents of phenolic, flavonoids, α-toc and ASA. Water stress also negatively effected the different yield attributes and seed nutrient quality. Foliar fertigation of wheat plants with α-Toc significantly improved the water stress tolerance of wheat plants in term of improvement in growth and seed yield associated with improved water relations, photosynthetic efficiency, contents of photosynthetic pigments and improvement in antioxidative defence mechanism (enzymatic and non-enzymatic antioxidants). Fertigation of water stressed wheat plants with α-Toc also improved the seed nutritional quality in terms of the contents of seed phenolics, flavonoids, activities of antioxidant enzymes and the content of α-, β- and γ-tocopherols. In conclusion, it was found that fertigation of water stressed wheat plants not only improved the water stress tolerance but also improved the seed yield and nutritional quality that will not only be helpful for the improvement in wheat yield that also be a step to solve the problem of malnutrition through the bio-fertification of α-Toc.

Effects of dietary astaxanthin and vitamin E and their interactions on the growth performance, pigmentation, digestive enzyme activity of kuruma shrimp (Marsupenaeus japonicus )

A 56-day feeding trial was done to investigate the interactive effects of astaxanthin (Ax) and vitamin E (α-Toc) on the performance of kuruma shrimp (Marsupenaeus japonicus). A 2 × 3 factorial experiment was conducted with six experimental diets containing two levels of Ax (0 and 0.6 g/kg diet [Ax0 and Ax0.6]) and three levels of α-Toc (0, 0.2 and 1 g/kg diet [α-Toc0, α-Toc0.2 and α-Toc1]). The results indicated that growth performance was significantly (p < 0.05) increased in shrimp fed with the Ax0.6 × α-Toc0.2 diet. Interactive effects between Ax and α-Toc on the growth parameters were observed. Furthermore, pigmentation performance was significantly (p < 0.05) better in the Ax0.6 groups. Interaction between Ax and α-Toc was also found in the Ax content of shrimp body parts. Interestingly, dietary α-Toc helped to reduce the Ax consumption rate, promote the absorption and increase the deposition of Ax in the muscle. Shrimps from the Ax0.6 groups showed significantly (p < 0.05) improved hepatopancreatic digestive enzyme activities compared with those of the Ax0 groups. Although no interactive (p > 0.05) effects were found between dietary α-Toc and Ax on total haemocyte count and tolerance against freshwater, dietary Ax and α-Toc supplementation showed better performance on these two parameters. It was concluded that dietary Ax and α-Toc functioned synergistically, and the shrimp fed with the diet containing 0.6 g Ax/kg diet Ax and 0.2 g α-Toc/kg diet showed improved growth and pigmentation performance compared with the other groups in the current study.

Impact of exogenously applied tocopherol on some key physio-biochemical and yield attributes in mungbean [Vigna radiata (L.) Wilczek] under limited irrigation regimes

The lipophilic vitamin E, α-Tocopherol (α-Toc), has been considered as a potent cellular antioxidant naturally occurring in biological membranes. It plays a number of key metabolic roles in plants exposed to various stressful cues. A field experimental was conducted on mungbean [Vigna radiata (L.) Wilczek] under varying water-limited regimes and the plants were sprayed with four levels of α-Toc (0, 100, 200, and 300 mg L−1) at the vegetative stage of plant growth. The results showed that limited field irrigation regimes (desiccated conditions) caused a marked reduction in growth parameters (shoot and root fresh and dry weights; shoot and root lengths), photosynthetic pigments (chlorophyll a and b), total soluble proteins (TSP) and yield attributes (number of pods, seeds per plant, weight of ripened pods and 100-seeds weight), while, in contrast, water deficiency induced an increase in phenolics, proline, glycine betaine (GB), hydrogen peroxide (H2O2), malondialdehyde (MDA), reducing and non-reducing sugars, total free amino acids, endogenous tocopherol levels, and activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). However, the foliar spray of α-Toc significantly improved shoot and root fresh and dry weights, shoot and root lengths, chlorophyll a and b contents, phenolics, proline, GB, reducing and non-reducing sugars, TSP, total free amino acids, endogenous tocopherol level, and activities of antioxidant enzymes (SOD, POD, and CAT), as well as yield parameters. On the other hand, it significantly decreased the MDA and H2O2 levels. Of all varying levels of α-Toc used, 100 mg L−1 was most effective in causing enhanced accumulation of ascorbic acid (AsA), MDA, and cellular tocopherols particularly in cv. Cyclone 7008, while proline and catalase in cv. Cyclone 8009. Of both cultivars, Cyclone 7008 was superior to the other cultivar in proline, TSP, SOD, and POD enzymes, but cv. Cyclone 8009 being superior in shoot fresh weights, root fresh weight, shoot and root lengths, chlorophyll a and b, phenolics, H2O2, AsA, MDA, CAT, number of pods, seeds, weight of ripened pods, and weight of 100 seeds under dry land conditions. Overall, exogenously applied tocopherol improved yield and myriad of key physio-biochemical attribute in mungbean.

Tocochromanols in wood: a potential new tool for dendrometabolomics.

Tocochromanols are the most abundant lipid-soluble antioxidants in plants. Among them, α-tocopherol (α-Toc) shows a particularly high sensitivity to environmental stressors and its content is used as a stress biomarker even in non-photosynthetic tissues. Nevertheless, the presence of tocochromanols has not been described yet in the xylem of woody plants, even when their functions regarding cell membrane protection and the transport of photoassimilates may be crucial in this tissue and despite its potential utility in dendrometabolomics. Considering all these, we aimed to determine the presence and distribution of tocochromanols in the xylem of woody plants, to examine their responsiveness to high temperature and to evaluate their potential as environmental bioindicators. The analysis of 29 phyllogenetically diverse species showed that α-Toc is the most abundant and frequent tocochromanol in the xylem and is ubiquitously present in all the studied species, with a concentration ranging from 0.5 to 39.3 μg g(-1) of dry weight. α-Tocopherol appeared to be mainly located in the parenchyma rays and was found in both the sapwood and the heartwood, suggesting that it is present even in dead parenchyma cells. The levels of α-Toc in the xylem did not change in response to locally induced xylem heating, but responded positively to the 3-year moving average of annual precipitation. The present findings suggest that α-Toc may be linked to changes in climatic stress. This should enhance further research on the environmental controls of α-Toc variation in the xylem as a first step towards a deeper understanding of dendrometabolomics.

Excessive α-tocopherol exacerbates microglial activation and brain injury caused by acute ischemic stroke.

The vitamin E family includes both tocopherols and tocotrienols, where α-tocopherol (αTOC) is the most bioavailable form. Clinical trials testing the therapeutic efficacy of high-dose αTOC against stroke have largely failed or reported negative outcomes when a "more is better" approach to supplementation (>400 IU/d) was used. This work addresses mechanisms by which supraphysiologic αTOC may contribute to stroke-induced brain injury. Ischemic stroke injury and the neuroinflammatory response were studied in tocopherol transfer protein-deficient mice maintained on a diet containing αTOC vitamin E at the equivalent human dose of 1680 IU/d. Ischemic stroke-induced brain injury was exacerbated in the presence of supraphysiologic brain αTOC levels. At 48 h after stroke, S100B and RAGE expression was increased in stroke-affected cortex of mice with elevated brain αTOC levels. Such increases were concomitant with aggravated microglial activation and neuroinflammatory signaling. A poststroke increase in markers of oxidative injury and neurodegeneration in the presence of elevated brain αTOC establish that at supraphysiologic levels, αTOC potentiates neuroinflammatory responses to acute ischemic stroke. Exacerbation of microglial activation by excessive αTOC likely depends on its unique cell signaling regulatory properties independent of antioxidant function. Against the background of clinical failure for high-dose αTOC, outcomes of this work identify risk for exacerbating stroke-induced brain injury as a result of supplementing diet with excessive levels of αTOC.

Abstract 34: Serum Dietary Antioxidants in Young Adulthood and Incident Cardiovascular Disease in Middle Age: The CARDIA Study

Introduction: Serum levels of carotenoids and tocopherols are objective measures of dietary intakes, including fruits and vegetables (carotenoids and the carotenoid index), nuts/seeds and vegetable oils (α-tocopherol (α-toc)), and oils, margarines, salad dressing and other related foods (γ-tocopherol (γ-toc)). Little research has examined these serum measures in young adulthood as predictors of the incidence of cardiovascular disease (CVD) in middle age. Hypothesis: Circulating carotenoids and tocopherols in young adulthood are inversely associated with incident CVD during middle-age. Methods: Participants from the Coronary Artery Risk Development in Young Adults study (CARDIA, N=5,115) with serum measures of carotenoids and tocopherols at year 0 of the study and concurrent risk factor data were included in the analysis (N=4,810, mean age 25). CVD events (n=167, coronary heart disease, stroke, and heart failure, adjudicated based on medical records) were ascertained over 25 years of follow up. Multivariable Poisson regression (PROC Genmod) was used to estimate the incidence density of CVD over 25 years according to quintiles of the carotenoid index (sum of 4 variables, defined in Table), α-toc and γ-toc with adjustment for demographic, lifestyle, and clinical covariates. Results: There was a strong inverse association between higher levels of the carotenoid index and incidence of CVD, a suggestive inverse association between higher levels of α-toc and incident CVD, and a positive association between γ-toc and incident CVD (Table). The carotenoid, lycopene, did not predict incident CVD. Conclusions: The results for the carotenoids and α-tocopherol underscore how a nutrient rich, plant based dietary pattern in young adulthood may inversely associate with CVD events through middle age. The divergent γ-toc results may reflect the kinetics of its metabolism, a poorer dietary pattern, or higher intake of a food from the food supply (e.g. margarine in 1985-6) that may increase risk.

Determination of α-Tocopherol in Vegetable Oils Using a Molecularly Imprinted Polymers–Surface-Enhanced Raman Spectroscopic Biosensor

We report the development of a novel hybrid "capture-detection" molecularly imprinted polymers-surface-enhanced Raman spectroscopic (MIPs-SERS) biosensor for the detection and quantification of α-tocopherol (α-Toc) in vegetable oils. α-Toc served as the template for MIPs synthesis. Methacrylic acid formed as the functional monomer. Ethylene glycol dimethacrylate was the cross-linking agent, and 2,2'-azobisisobutyronitrile was used as the initiator. The synthesized MIPs functioned to rapidly and selectively adsorb and separate α-Toc from oil components. We validated a dendritic silver nanostructure synthesized by a displacement reaction to be a suitable SERS substrate for the enhancement of Raman signals. Second-derivative transformations and chemometric models based upon SERS spectral features confirmed the possibility of a rapid and precise detection and quantification of different spiking levels of α-Toc in four different sources of vegetable oils (Mahalanobis distance from 15.93 to 34.01 for PCA model; R > 0.92, RMSE < 0.41 for PLSR model). The MIPs-SERS biosensor had a high sensitivity as well as a good recovery for α-Toc analysis in vegetable oils. The entire analysis required 15 min or less to complete with limited sample preparation.

Use of alpha-tocopherol esters for topical vitamin E treatment: evaluation of their skin permeation and metabolism

The aim of this work was to investigate new pro-vitamins based on α-tocopherol (α-Toc) and fatty acids, and to compare their properties with those of α-tocopherol acetate (α-TAc). Skin levels of α-Toc-fatty acid ester conjugates, total α-Toc and endogenous α-Toc were measured in skin samples taken from separate groups of treated and untreated rats. Multiple and extensive treatment with α-Toc oleate and α-TAc was also carried out to assess the skin accumulation and safety of these esters. The in-vivo studies revealed that α-Toc-fatty acid conjugates penetrated into the skin quantitatively while being comparable with the permeation of α-TAc. Differences were found between the levels of total α-Toc and endogenous α-Toc after application of α-TAc, α-Toc oleate, α-Toc linoleate, α-Toc-α linolenate and α-Toc palmitate, indicating that α-Toc conjugates of these fatty acids, but not α-Toc γ-linolenate or α-Toc stearate, were hydrolysed to free α-Toc. In long-term and extensive treatment, α-TAc was found to be lethal to rats treated with 1.15 mg/kg of this agent, which had been spread over 16 cm(2) of skin. Similar treatment with α-Toc oleate did not produce any side effects. This study suggests that α-Toc conjugates with unsaturated fatty acids may be a good alternative as stable vitamin E derivatives, rather than the α-TAc ester.

Effects of Monascus-fermented grain extracts on plasma antioxidant status and tissue levels of ubiquinones and α-tocopherol in hyperlipidemic rats

We investigated the effects of Monascus-fermented mixed grain extracts (MFGEs) enriched with bioactive mevinolins (natural statins) and coenzyme Qs (CoQ9+CoQ10) on the blood lipids, antioxidant status, and tissue levels of CoQs and α-tocopherol (α-Toc) in hyperlipidemic rats. The oral administration of MFGEs (300mg/kg body weight per day) for 8weeks resulted in a significant decrease in blood levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and LDL-C/high-density lipoprotein cholesterol (HDL-C) ratio compared to the control and lovastatin supplement group of a dosage of 20mg/kg per day (p<0.05). Furthermore, a significant increase in the ratios of α-Toc/LDL-C and CoQs/LDL-C in plasma and tissues and improvement in plasma antioxidant status as measured by TBARS and TRAP were observed in hypercholesterolemic rats (p<0.05). Regarding the effects of MFGEs on antioxidant levels of plasma and tissues, there were significant increases in the levels of α-Toc (p<0.05) and CoQs (p<0.01) after the 8-week MFGEs treatment. These data indicate that MFGEs supplementation not only decreases blood lipids and lipid peroxidation but also increases levels of antioxidants such as α-Toc and CoQs and may improve plasma antioxidant status as well as a hypolipidemic effect.

Plasma and Tissue Concentrations of α-Tocopherol and δ-Tocopherol Following High Dose Dietary Supplementation in Mice

Vitamin E isoforms are essential nutrients that are widely used as dietary supplements and therapeutic agents for a variety of diseases. However, their pharmacokinetic (PK) properties remain poorly characterized, and high dosage animal studies may provide further information on their in vivo functions and pharmacological effects. In this study, alpha-tocopherol (α-toc) and delta-tocopherol (δ-toc) levels were measured in mouse plasma and tissues following their high dosage dietary supplementation. Average α-toc levels at 5, 10 and 20 g α-toc/kg diet increased over baseline levels 6-fold in plasma, 1.6-fold in brain, and 4.9-fold in liver. These elevated α-toc concentrations remained constant from 5 to 20 g α-toc/kg diet, rather than showing further increases across these dosages. No α-toc-related toxicity occurred at these high dosages, and strain-specific differences in liver and brain α-toc levels between Balb/cJ and C57Bl/6J mice were observed. Relatively high-dosage administration of dietary δ-toc for 1 or 4 weeks resulted in 6–30-fold increases in plasma and liver levels between dosages of 0.33 and 1.67 g δ-toc/kg diet. Co-administration of sesamin with δ-toc further increased δ-toc levels between 1.3- and 14-fold in plasma, liver, and brain. These results provide valuable PK information on high dosage α-toc and δ-toc in mouse and show that supplementation of sesamin with δ-toc further increases δ-toc levels over those seen with δ-toc supplementation alone.

Elevated Plasma γ-Tocopherol and Decreased α-Tocopherol in Men Are Associated With Inflammatory Markers and Decreased Plasma 25-OH Vitamin D

Chronic inflammation is a risk factor for many diseases of aging. Endogenous oxidants are thought to mediate the effects of inflammation and γ-Tocopherol (γ-Toc) may mitigate damage from nitrogen-based oxidants; however, no physiological requirement for γ-Toc has been established. Regulation of tocopherols and their functional significance are poorly defined, thereby limiting their application in prevention. Using stored plasma samples from 657 male control subjects in a previous study of prostate cancer, we have analyzed associations of the tocopherols, inflammation markers, and 25-hydroxy (OH) vitamin D. Plasma α-Toc and γ-Toc were inversely correlated, whereas δ-Toc and α-Toc levels were positively correlated, suggesting a unique regulatory mechanism. γ-Toc levels were positively and α-Toc negatively associated with plasma C-reactive protein (CRP) and urinary isoprostane F2t, which are markers of inflammation and oxidation. Ethnic variability in tocopherols was observed; however, this may be explained by differences in plasma 25-OH vitamin D, as γ-Toc levels varied inversely and α-Toc positively with 25-OH vitamin D. In these data, all-cause mortality appeared to be positively associated with CRP and inversely with 25-OH vitamin D. We hypothesize that plasma levels of tocopherols may serve as markers of systemic inflammation, complicating epidemiologic assessment of their role in cancer etiology.

A New Wrinkle on Topical Vitamin E and Photo-inflammation: Mechanistic Studies of a Hydrophilic γ-Tocopherol Derivative Compared with α-Tocopherol

The antioxidant function of vitamin E is thought to mediate its photo-protective effects. Cyclooxygenase-2 (COX-2) is an important mediator of early photo-inflammation. Thus, the ability of gamma-tocopherol to inhibit COX-2 activity independently of its antioxidant function raises important questions regarding potential roles that this form of vitamin E plays in photo-protection and skin cancer chemoprevention.

Metabolic effects of enteral versus parenteral alanyl‐glutamine (AG) dipeptide in critically ill tube‐fed patients

Enteral and parenteral glutamine (Gln) supplementation is being investigated in critically ill patients, but metabolic effects of different routes of administration are unknown. Methods This double-blind, randomized study compared enteral versus parenteral AG (EN-AG vs IV-AG) on plasma Gln, plasma antioxidants, intestinal permeability to lactulose/mannitol and nitrogen (N) balance over a 8-day period in 44 ICU patients requiring tube feeding. Subjects each received an identical, Gln-free, isocaloric tube feeding 24 h/day and randomized to control (no AG, n=12), EN-AG (n=16) or IV-AG (n=16) groups (AG at 0.5 g/kg/day). Results APACHE II scores were similar at entry in all groups as was energy and N delivery during study. Plasma Gln rose significantly in the IV-AG group. EN-AG subjects showed a significant increase in α-tocopherol (α-toc) levels over time (day 1, 1155±105 vs day 9, 1272±133 μg/dL; p=0.019) and maintained γ-tocopherol (γ-toc) levels. In contrast, α-toc levels were unchanged and γ-toc levels fell significantly in IV-AG and control groups. There were no differences between groups in levels of vitamin C and glutathione, intestinal permeability or N balance. In conclusion, this study shows that enteral Gln may maintain or increase plasma tocopherols in ICU patients. Route of Gln administration does not influence plasma vitamin C or glutathione levels, gut permeability or N balance in ICU patients over an 8-day period. Supported by NIH RO3 DK54823.

Oleanolic Acid Protects against Myocardial Ischemia-Reperfusion Injury by Enhancing Mitochondrial Antioxidant Mechanism Mediated by Glutathione and α-Tocopherol in Rats

The effect of oleanolic acid (OA) pretreatment on myocardial ischemia-reperfusion (I-R) injury was investigated using an ex vivo rat heart model. Pretreatment with OA at daily doses (0.6 and 1.2 mmol/kg) for 3 days significantly protected against I-R injury in isolated rat hearts, as evidenced by the decrease in the extent of lactate dehydrogenase (LDH) leakage and improvement in contractile force recovery. The cardioprotection was associated with a slight increase in mitochondrial reduced glutathione (GSH) level and a significant increase in mitochondrial alpha-tocopherol (alpha-TOC) level, when compared with the unpretreated I-R group. To further investigate the mechanism of myocardial protection, pretreatment with a single dose of OA (1.2 mmol/kg) produced a time-dependent protection against myocardial I-R injury as assessed by LDH leakage, with the maximum extent of protection occurring at 48 hour post-dosing. The maximum cardioprotection was associated with parallel increases in mitochondrial GSH and alpha-TOC levels in ischemic-reperfused hearts, with the stimulation of the alpha-TOC level being optimal. Furthermore, buthionine sulfoximine/phorone (BSO/PHO) treatment, while abolishing the enhancing effect of OA on mitochondrial GSH, did not completely abrogate the cardioprotection against I-R injury. The remnant cardioprotection was associated with an increase in mitochondrial alpha-TOC level, when compared with the unpretreated I-R group with BSO/PHO. The results suggest that the cardioprotection afforded by OA pretreatment against I-R injury may at least in part be attributed to the enhancement of mitochondrial antioxidant mechanism mediated by GSH and alpha-TOC, particularly under I-R conditions. Abbreviations. BSO:buthionine sulfoximine GSH:reduced glutathione I-R:ischemia-reperfusion alpha-LA:alpha-lipoic acid LDH:lactate dehydrogenase OA:oleanolic acid PHO:phorone alpha-TOC:alpha-tocopherol.

Determination of tissue α-tocopherol in senescence-accelerated mice by high-performance liquid chromatography with fluorimetric detection

α-Tocopherol (α-Toc) plays an important role in various distinct physiological functions. A convenient assay using high-performance liquid chromatography with fluorimetric detection (the excitation and emission wavelength was 296 and 340 nm, respectively) was developed for the accurate determination of the α-Toc in biological samples. Methanol (1/7; v/v) was used to extract α-Toc in tissue samples by precipitating the proteins. The assay obtained a recovery rate between 97.3 and 104.2% by adding pure α-Toc to mice liver homogenate. The R.S.D. for intra-day ( n=7) and inter-day ( n=6, three consecutive days) analysis was 0.7–2.8 and 0.5–3.7%, respectively. The effect of age on the α-Toc levels in the liver, brain and heart homogenates of senescence-accelerated mice (SAM) was also studied. The α-Toc levels in the tissue homogenates increased gradually with age for both male and female mice, while the female had higher α-Toc levels than the males of the same age.

Effect of dietary vitamin E supplementation on the colour and lipid stability of fresh, frozen and vacuum-packaged beef

The effects of dietary vitamin E supplementation on tissue α-tocopherol (α-Toc) levels and on the susceptibility of fresh, frozen and vacuum-packaged beef to lipid oxidation and colour deterioration were investigated. Friesian cattle were fed diets containing 20 (basal, n=5) or 2000 (supplemented, n=5) IU (α-tocopheryl acetate/kg feed/day for approximately 50 days prior to slaughter. α-Toc levels were higher ( p < 0.05) in muscles from supplemented animals than from those on a basal diet. Significant differences in α-Toc levels were also observed between muscles from different treatment groups, the order of the supplemented group was: M. psoas major (PM) > M. longissimus dorsi (LD) > M. gluteus medius (GM) ( p < 0.05), and in the basal group the order was: PM > GM > LD ( p < 0.05). Supplemented fresh, frozen and vacuum packed beef showed greater colour and lipid oxidative stability than meat from the basal group after 7 days retail display at 4°C ( p < 0.05). Thus, dietary (α-Toc supplementation appeared to retard metmyoglobin and TBARS formation in LD, GM and PM and increased the colour shelf life of these muscles.

Protective effects of coenzyme Q10 and α-tocopherol against free radical-mediated liver cell injury.

In an attempt to provide further confirmation of the antioxidant role of reduced form of coenzyme Q homologue (CoQnH2) and α-tocopherol (α-Toc), we incubated isolated rat hepatocytes with a water-soluble radical initiator, 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH) in the presence or absence of exogenously added coenzyme Q10 (CoQ10) or α-Toc for 3 h at 37°C under an atmosphere of 95% oxygen and 5% carbon dioxide. In the control experiment without adding AAPH it was confirmed that added CoQ10 and α-Toc were incorporated into the cells and some CoQ10 were converted to CoQ10H2. Incubation of hepatocytes with 50 mM AAPH resulted in the formation of thiobarbituric acid-reactive substances and the decrease in cell viability and both were inhibited by exogenously added CoQ10 or α-Toc in a dose-dependent manner. The decrease in endogenous CoQ9H2 and α-Toc levels was observed by the addition of AAPH. Addition of CoQ10 inhibited the oxidation of CoQ9H2 to CoQ9 dose-dependently while the addition of α-Toc did not. These data suggest that both CoQnH2 and α-Toc act as antioxidants and can inhibit free radical-mediated cell injury.

乳房炎罹患牛3症例におけるα-トコフェロール, セレニウムおよびその関連酵素の変動

The serum levels of lipids, α-tocopherol (α-Toc) and selenium (Se), and the behavior of related enzymes were investigated during the nonlactational and lactational periods of cows (Cases 1, 2 and 3) with mastitis and of normal controls that developed no clinical abnormalities on the same farm. The serum levels of α-Toc in Cases 1 and 2, which lost their lactational function, decreased during the nonlactational period, compared to the controls. This tendency was particularly significant in Case 2. No significant difference was found in serum α-Toc between Case 3 and the controls. The serum level of Se decreased and blood glutathione peroxidase activity diminished during the nonlactational period of Case 1, whereas serum Se in cases 2 and 3 showed almost the same pattern of change as in the controls throughout the course of observation.The serum superoxide dismutase (SOD) activity diminished during the observation period in Cases 1, 2 and 3 compared to the controls. However, no significant differences were demonstrated in the serum levels of Cu and Zn (constitutive microquantity-elements of SOD) between cows with mastitis and the controls.The serum levels of total cholesterol decreased during the nonlactational period of Cases 1 and 2, accordinglythe serum phospholipids/total cholesterol ratio increased in these cows. On the other hand, the serum total cholesterol level and phospholipids/total cholesterol ratio in Case 3 showed no significant difference with the controls. The serum total bile acids in Cases 1, 2 and 3 showed no significant difference with the controls during the nonlactational period.This case was diagnosed that it case of 1, 2, unequilibrium of mammary glandular cell membran was induced in the serum table of α-Toc diminished in the reason of loss appetite during the nonlactational period.

718 ABNORMALITIES IN FAT-SOLUBLE VITAMIN STATUS AT THE TIME OF DIAGNOSIS IN INFANTS WITH CYSTIC FIBROSIS IDENTIFIED BY NEWBORN SCREEN

We have previously reported evidence of early protein malnutrition in some infants with cystic fibrosis (CF) identified by the newborn screening method using serum immunoreactive trypsinogen. We now report very early abnormalities in the status of fat-soluble vitamins in 6 of 10 infants (mean age, 5.7 weeks; range, 3-8 weeks) with CF identified by screening in 1984. Plasma retinol (PR) and γ and α-tocopherol (α-Toc) levels were measured by HPLC. Retinol binding protein (RBP) levels were measured by radial immunodiffusion. Total serum lipids, serum albumin, calcium, and phosphorus were measured by standard methods. Four infants had low levels of serum albumin (<3.2 g/dL), low levels of α-Toc (<3.0 μg/ml), and low total tocopherols/total serum lipid ratios (<0.6 mg/gm). Three of these infants had RBP levels of 1.5 mg/dl (normal, 1.4-3.3 mg/dL). PR levels were low in 5 infants (<20 μg/dL). PIVKA-II assays (by plasma immunoelectropheresis before and after BaCC3 absorption, representing noncarboxylated prothrombin specific for Vitamin K deficiency) were negative in all infants. Serum calcium and phosphorus levels were not low, suggesting adequate Vitamin D status. We conclude that as early as 6 weeks of age, some infants with CF have biochemical deficiencies of the fat-soluble vitamins A and E. However, these infants, all of whom had parenteral Vitamin K prophylaxis at birth, were not Vitamin K deficient.