Selective excretion of yolk-derived tocotrienols into the bile of the chick embryo

The aim of this study was to investigate the possibility of biodiscrimination between different forms of vitamin E during the development of the chick embryo. The vitamin E present in the initial yolk consisted of α-tocopherol (90%), (β+γ)-tocopherol (8%), α-tocotrienol (0.3%) and (β+γ)-tocotrienol (1.3%). In marked contrast, the vitamin E recovered from the bile of the day-16 embryo contained much higher proportions of α-tocotrienol (10%) and especially of (β+γ)-tocotrienol (42%). By the time of hatching, 56% of the vitamin E present in the bile was in the form of (β+γ)-tocotrienol. The residual yolk of the newly-hatched chick contained far greater proportions of α-tocotrienol (2.6%) and (β+γ)-tocotrienol (10%) than were present in the initial yolk. The results suggest that the liver of the embryo may selectively excrete tocotrienols as components of bile, whilst retaining the tocopherols within the hepatocytes. The increased proportions of tocotrienols in the residual yolk may result from the recycling of bile from the gall bladder to the yolk. The liver of the day-old chick contained α-tocopherol as the main form of vitamin E (90%) with only a small proportion (0.2%) of (β+γ)-tocotrienol. The α-tocopherol form was also the main vitamin E component in the brain (85%), heart (79%), lung (82%) and adipose tissue (91%) of the day-old chick. The present study suggests the occurrence of a high degree of biodiscrimination between tocopherols and tocotrienols during the development of the chick embryo.     

Embryonic development within carotenoid-enriched eggs influences the post-hatch carotenoid status of the chicken

Carotenoids in the diet of the laying hen are incorporated into the egg yolk and subsequently into the liver and other tissues of the chicken embryo. Since these pigments are known to provide a range of health benefits to a variety of animals, it is of interest to know whether the effects of maternally derived carotenoids are strictly limited to the embryonic period or if they persist in the progeny after hatching. The aim of this study is to compare the effectiveness of pre-hatch (from the hen's diet) with that of post-hatch (from the progeny's diet) supplementation with carotenoids on the carotenoid status of the chick during the first 4 weeks of post-hatch life. Hens were fed a control diet or a diet supplemented with a carotenoid-rich extract of alfalfa. Eggs from the supplemented hens contained up to 22 times more carotenoids than the controls. The concentration of carotenoids in the livers of chicks hatching from the enriched eggs was initially 29 times greater than in the control chicks. Hepatic carotenoid concentrations in chicks from enriched eggs maintained post-hatch on the control diet were sustained at higher values compared with chicks from control eggs that were fed post-hatch on the carotenoid-supplemented diet, for at least the first 7 days. However, by 14 days, the latter group had overtaken the former in terms of liver carotenoid levels. Thus, under these conditions, maternal effects predominate for at least the first week after hatching, whereas from 2 weeks onwards, the progeny's diet becomes the main determinant of its carotenoid status. Since the antioxidant and immunostimulatory roles of carotenoids are likely to be especially important during the immediate post-hatch period, maternal dietary intake of carotenoids may have important ramifications for the viability of the offspring.     

Significance of chick quality score in broiler production

The quality of day old chicks is crucial for profitable broiler production, but a difficult trait to define. In research, both qualitative and quantitative measures are used with variable predictive value for subsequent performance. In hatchery practice, chick quality is judged on a binomial scale, as chicks are divided into first grade (Q1-saleable) and second grade (Q2) chicks right after hatch. Incidences and reasons for classifying chicks as Q2, and potential of these chicks for survival and post-hatch performance have hardly been investigated, but may provide information for flock performance. We conducted an experiment to investigate (1) the quality of a broiler flock and the relation with post-hatch flock performance based on a qualitative score (Pasgar©score) of Q1 chicks and based on the incidence of Q2 chicks and (2) the reasons for classifying chicks as Q2, and the potential of these chicks for survival and post-hatch growth. The performance was followed of Q1 and Q2 chicks obtained from two breeder flocks that hatched in two different hatching systems (a traditional hatcher or a combined hatching and brooding system, named Patio). Eggs were incubated until embryo day 18, when they were transferred to one of the two hatching systems. At embryo day 21/post-hatch day 0, all chicks from the hatcher (including Q2 chicks) were brought to Patio, where the hatchery manager marked the Q2 chicks from both flocks and hatching systems and registered apparent reasons for classifying these chicks as Q2. Chick quality was assessed of 100 Q1 chicks from each flock and hatching system. Weights of all chicks were determined at days 0, 7, 21 and 42. There were no correlations between mean Pasgar©score and post-hatch growth or mortality, and suboptimal navel quality was the only quality trait associated with lower post-hatch growth. Growth was clearly affected by breeder flock and hatching system, which could not be linked to mean Pasgar©score or incidence of Q2 chicks. Q2 chicks showed lower post-hatch growth compared to Q1 chicks but effects on flock performance at slaughter weight were limited because early mortality in Q2 chicks was high (62.50% at 7 days). We concluded that chick qualitative scores and the incidence of Q2 chicks may be informative for the quality of incubation, but are not predictive for post-hatch flock performance. Culling Q2 chicks after hatch is well-founded in terms of both animal welfare and profitability.     

Natural forms of vitamin E: metabolism,antioxidant, and anti-inflammatory activities and their role in disease prevention and therapy

The vitamin E family consists of four tocopherols and four tocotrienols. α-Tocopherol (αT) is the predominant form of vitamin E in tissues and its deficiency leads to ataxia in humans. However, results from many clinical studies do not support a protective role of αT in disease prevention in people with adequate nutrient status. On the other hand, recent mechanistic studies indicate that other forms of vitamin E, such as γ-tocopherol (γT), δ-tocopherol, and γ-tocotrienol, have unique antioxidant and anti-inflammatory properties that are superior to those of αT in prevention and therapy against chronic diseases. These vitamin E forms scavenge reactive nitrogen species, inhibit cyclooxygenase- and 5-lipoxygenase-catalyzed eicosanoids, and suppress proinflammatory signaling such as NF-κB and STAT3/6. Unlike αT, other vitamin E forms are significantly metabolized to carboxychromanols via cytochrome P450-initiated side-chain ω-oxidation. Long-chain carboxychromanols, especially 13′-carboxychromanols, are shown to have stronger anti-inflammatory effects than unmetabolized vitamins and may therefore contribute to the beneficial effects of vitamin E forms in vivo. Consistent with mechanistic findings, animal and human studies show that γT and tocotrienols may be useful against inflammation-associated diseases. This review focuses on non-αT forms of vitamin E with respect to their metabolism, anti-inflammatory effects and mechanisms, and in vivo efficacy in preclinical models as well as human clinical intervention studies.     

Vitamin A metabolism: α-Tocopherol modulates tissue retinol levels in vivo,and retinyl palmitate hydrolysis in vitro

The steady-state concentrations of retinol in rat tissues varied as a function of dietary α-tocopherol. The liver, kidney, and intestinal retinol concentrations increased in animals fed an α-tocopherol-deficient diet despite a decrease (liver) or no change (kidney and intestine) in the concentrations of total vitamin A. In contrast, in lung the concentrations of both retinol and total vitamin A decreased. α-Tocopherol inhibited retinyl palmitate hydrolase in vitro in liver, kidney, and intestine; had minimal effect on the testes hydrolase; and stimulated the lung hydrolase. Fifty percent inhibition of the liver hydrolase was provided by an α-tocopherol concentration (100 μm), close to that reported in livers of rats fed a purified diet, constituted with moderately low amounts of α-tocopheryl acetate. Phylloquinone (vitamin K₁) inhibited the retinyl palmitate hydrolase in vitro in all tissues tested, and was about fivefold more potent than α-tocopherol. The effects of phylloquinone and α-tocopherol on the liver hydrolase were additive, not synergistic. The antioxidant N,N′-diphenyl-p-phenylenediamine, the most effective synthetic vitamin E substitute known, had little effect on the hydrolase. These data show that α-tocopherol effects vitamin A metabolism in several tissues, and suggest that it may be a physiological effector of tissue retinol homeostasis.     

Increased ��-tocotrienol content in seeds of transgenic rice overexpressing Arabidopsis ��-tocopherol methyltransferase

Vitamin E comprises a group of eight lipid soluble antioxidant compounds that are an essential part of the human diet. The ??-isomers of both tocopherol and tocotrienol are generally considered to have the highest antioxidant activities. ??-tocopherol methyltransferase (??-TMT) catalyzes the final step in vitamin E biosynthesis, the methylation of ??- and ??-isomers to ??- and ??-isomers. In present study, the Arabidopsis ??-TMT (AtTMT) cDNA was overexpressed constitutively or in the endosperm of the elite japonica rice cultivar Wuyujing 3 (WY3) by Agrobacterium-mediated transformation. HPLC analysis showed that, in brown rice of the wild type or transgenic controls with empty vector, the ??-/??-tocotrienol ratio was only 0.7, much lower than that for tocopherol (~19.0). In transgenic rice overexpressing AtTMT driven by the constitutive Ubi promoter, most of the ??-isomers were converted to ??-isomers, especially the ??- and ??-tocotrienol levels were dramatically decreased. As a result, the ??-tocotrienol content was greatly increased in the transgenic seeds. Similarly, over-expression of AtTMT in the endosperm also resulted in an increase in the ??-tocotrienol content. The results showed that the ??-/??-tocopherol ratio also increased in the transgenic seeds, but there was no significant effect on ??-tocopherol level, which may reflect the fact that ??-tocopherol is present in very small amounts in wild type rice seeds. AtTMT overexpression had no effect on the absolute total content of either tocopherols or tocotrienols. Taken together, these results are the first demonstration that the overexpression of a foreign ??-TMT significantly shift the tocotrienol synthesis in rice, which is one of the world??s most important food crops.     

Unleashing the untold and misunderstood observations on vitamin E

Paradoxically, meta-analysis of human randomized controlled trials revealed that natural but not synthetic α-tocopherol supplementation significantly increases all-cause mortality at 95% confidence interval. The root cause was that natural α-tocopherol supplementation significantly depressed bioavailability of other forms of vitamin E that have better chemo-prevention capability. Meta-analysis outcome demonstrated flaws in the understanding of vitamin E. Reinterpretation of reported data provides plausible explanations to several important observations. While α-tocopherol is almost exclusively secreted in chylomicrons, enterocytes secrete tocotrienols in both chylomicrons and small high-density lipoproteins. Vitamin E secreted in chylomicrons is discriminately repacked by α-tocopherol transfer protein into nascent very low-density lipoproteins in the liver. Circulating very low-density lipoproteins undergo delipidation to form intermediate-density lipoproteins and low-density lipoproteins. Uptake of vitamin E in intermediate-density lipoproteins and low-density lipoproteins takes place at various tissues via low-density lipoproteins receptor-mediated endocytosis. Small high-density lipoproteins can deliver tocotrienols upon maturation to peripheral tissues independent of α-tocopherol transfer protein action, and uptake of vitamin E takes place at selective tissues by scavenger receptor-mediated direct vitamin E uptake. Dual absorption pathways for tocotrienols are consistent with human and animal studies. α-Tocopherol depresses the bioavailability of α-tocotrienol and has antagonistic effect on tocotrienols in chemo-prevention against degenerative diseases. Therefore, it is an undesirable component for chemo-prevention. Future research directions should be focused on tocotrienols, preferably free from α-tocopherol, for optimum chemo-prevention and benefits to mankind.     

Incubation and hatch management: consequences for bone mineralization in Cobb 500 meat chickens

From ~35 days of age fast growing meat chickens spend extended periods sitting or lying and less time standing. In a fast-feathering parent line lower early incubation temperatures which delayed chick hatch time, improved bone ash and extended their standing time. This incubation study assessed the consequences of incubation temperatures, hatch time and chick management at hatch/take off on femoral bone ash (BA) in Cobb 500 meat chickens. Embryos were incubated under either Control (between 37.8°C and 38.2°C egg shell temperature (EST)) or a Slow start (from 37.2°C at sett (the start of incubation), reaching 37.8°C EST at day 13 incubation), temperatures. Hatched chicks were identified at 492 h (20.5 days of incubation – classified as early (E)) or, between >492 and ⩽516 h (>20.5 and ⩽21.5 days of incubation – classified as late (L)), from setting. The E hatch chicks were allocated across three post-hatch treatments; treatment 1: E hatch chicks that were sampled E at 492 h from setting; treatment 2: E hatch chicks that were fed for a further 24 h in a floorpen before being sampled L at 516 h from setting; treatment 3: E hatch chicks that spent a further 24 h in the incubator before being sampled L at 516 h from setting. All L hatch chicks formed one treatment group which was sampled L at 516 h (i.e. L hatch chicks sampled L). It is not possible to sample L hatching chicks E hence this treatment is absent from the experimental design. Slow start incubation resulted in a higher total hatch percentage with a greater proportion of chicks hatching L, compared with the Control incubation. The L hatching chicks had significantly higher BA than the E hatching chicks. Of the E hatching chicks, those sampled both E and L had significantly lower BA than E hatching chicks fed for 24 h before L sampling. The E hatch, fed and sampled L chicks had the numerically highest BA, which was not significantly different from the BA of the L hatching chicks sampled L These results demonstrate that BA at hatch can be improved, either by extending the incubation period through a Slow start incubation profile, inducing L hatch, or alternatively, via the prompt provision of feed to E hatching chicks.     

Response of Whole Blood,Erythrocyte and Plasma Vitamin E Content to Dietary Vitamin E Intake in the Chick

Whole blood, red blood cells (RBC), and plasma vitamin E (VE) levels in chicks fed dietary VE (dl-α-tocopheryl acetate, dl-αTa) supplementation in steps of 0.0, 5.0, 10.0, 15.0, 20.0 and 30.0 mg/Kg were determined to examine their usefulness as an index of VE status. The increase in VE level was significant and linear in whole blood (r = 0.90), RBC (r = 0.89) and plasma (r = 0.93) in response to dietary VE intake. There was a close correlation between VE in plasma vs whole blood (r = 0.90), plasma vs RBC (r = 0.91) and whole blood vs RBC (r = 0.95). The plasma VE content was 1.2–1.8 times greater than that of whole blood, and 6.6–12.5 times greater than that of RBC. The plasma total lipids content was not affected by the dietary VE intake, whereas the level of VE in the plasma total lipids was significantly increased with increasing supplementation. Alpha tocopherol was the major isomer (ca 92 %) of VE in whole blood, RBC and plasma at hatching. The small proportions of β-tocopherol (ca 2 %), γ-tocopherol (ca 5 %) and α-tocotrienol (ca 1 %) observed at 1 day of age had decreased or totally disappeared by 7 days of age after feeding the VE-free basal diet. The data showed that in the chick, the whole blood and RBC levels of VE were as sensitive and reliable indexes of dietary VE status as was that of the plasma.     

Vitamin E,glutathione S-transferase and γ-glutamyl transpeptidase activities in cultured hepatocytes of rats treated with carcinogens

• 1.1. The effects of α-tocopherol and γ-tocotrienol on glutathione S-transferase (GST) and γ-glutamyl transpeptidase (γ-GT) activities in cultured hepatocytes prepared from rats treated with diethylnitrosamine (DEN) and 2-acetylaminofluorene (AAF) were investigated.
• 2.2. Both the α-tocopherol and γ-tocotrienol treated hepatocytes showed significantly higher (P < 0.05) GST activities than untreated hepatocytes prepared from the carcinogen treated rats in the first 3 days of culture. Treatment with α-tocopherol and γ-tocotrienol generally resulted in a tendency to increase the GST activities above that in the untreated hepatocytes.
• 3.3. Treatment with high doses (125–250 μM) of α-tocopherol and low doses (12.5–25 μM) of γ-tocotrienol generally resulted in a significant reduction in γ-GT activities at 1–3 days. γ-GT activities are reduced as the dose of α-tocopherol and γ-tocotrienol are increased.

     

Effects of supplementation with vitamin E on the performance and the tissue peroxidation of broiler chicks and the stability of thigh meat against oxidative deterioration

Two experiments were conducted: Expt 1 determined the optimal allowance of vitamin E in the diet for broiler chicks aged 0–3 weeks; Expt 2 investigated the effects of different dietary levels of vitamin E (α-tocopherol) on the performance and the oxidative stability of thigh meat of broiler chicks during storage. In Expt 1, 1-day-old 900 broiler chicks were allocated to five treatments, each with six replicates (cages) of 22 as-hatched chicks for performance evaluation, and another cage of 45 male chicks for determining plasma and hepatic α-tocopherol and thiobarbituric acid reactive substances (TBARS) concentration in blood and liver. The basal dietary α-tocopherol concentration was 13 mg/kg, and the five α-tocopherol acetate supplementation levels were 0, 5, 10, 50 and 100 mg/kg. For 0–3-week-old broiler chicks fed with maize–soya bean meal–soya oil type diet, supplementation of vitamin E did not influence the feed intake, but tended to improve growth and feed utilization, however there was no significant correlation between performance and vitamin E supplementation level. Significant positive correlations existed between dietary supplemental vitamin E level and plasma or hepatic α-tocopherol concentrations (P<0.05), and a negative correlation with hepatic TBARS levels no matter at what age (11, 16 and 21 days). In Expt 2, 2200 broiler chicks were randomly allocated to five treatments with four replicates (pens) in each. Chicks were fed ad libitum five pellet diets supplemented with vitamin E at 5, 10, 20, 50 and 100 mg/kg of diet, respectively. The basal dietary α-tocopherol level of grower and finisher diets were 7 and 6 mg/kg, respectively. Supplementation of vitamin E tended to improve growth and feed utilization of birds during 0–3 weeks of age, but the performance from 0 to 6 weeks of age were not influenced. The hepatic α-tocopherol concentrations of 6-week-old chicks linearly increased with the dietary vitamin E levels (R²=0.98, P<0.001). The content of TBARS in the thigh meat over 4 days of storage under 4°C was significantly decreased by increasing dietary vitamin E level (P<0.05). There was a significant inverse relationship between TBARS value in the thigh meat and the dietary vitamin E level (R²=0.93, P<0.01). Supplementation of vitamin E significantly improved the meat quality stability substantially against oxidative deterioration. Comparing the hepatic α-tocopherol levels of chicks in Expts 1 and 2, total allowance of dietary α-tocopherol of 20–30 mg/kg could sustain relatively constant hepatic α-tocopherol level at round about 2–2.5 μg/kg.     

High dietary vitamin A interferes with tissue α-tocopherol concentrations in fattening pigs: a study that examines administration and withdrawal times

This study aimed to assess the interaction between different dietary vitamin A (dVitA) levels and the same concentration of vitamin E (100 IU all-rac-α-tocopheryl acetate/kg feed) in growing-finishing pigs. In the first experiment, two fat sources × two dVitA levels (0 v. 100 000 IU) were used. The supplementation of 100 000 IU dVitA induced a range of 5.13 to 30.03 μg retinol/g liver, 62.78 to 426.88 μg retinol palmitate/g liver, and 0.60 to 1.96 μg retinol/g fat. Dietary fat did not affect retinol or retinyl palmitate deposition in pigs. The high concentration of dVitA produced lower fat and liver α-tocopherol concentrations, and increased susceptibility of muscle tissue to oxidation. A second experiment was carried out to study the retinol and α-tocopherol retention at different withdrawal times prior to slaughter (two dVitA levels; 0 v. 100 000 IU). A high dose of 100 000 IU vitamin A during a short 2-week period was enough to induce α-tocopherol depletion in liver and fat to a similar extent as when 100 000 IU were administered during the whole fattening. Muscle, fat and liver α-tocopherol concentrations were not affected by dVitA in the 1300-13 000 IU/kg range, but liver α-tocopherol concentration was higher when vitamin A was removed from the vitamin mix 5 weeks prior to slaughter (experiment 3).     

Disruption of mouse cytochrome p450 4f14 (Cyp4f14 gene) causes severe perturbations in vitamin E metabolism

Vitamin E is a family of naturally occurring and structurally related lipophilic antioxidants, one of which, α-tocopherol (α-TOH), selectively accumulates in vertebrate tissues. The ω-hydroxylase cytochrome P450-4F2 (CYP4F2) is the only human enzyme shown to metabolize vitamin E. Using cDNA cloning, cell culture expression, and activity assays, we identified Cyp4f14 as a functional murine ortholog of CYP4F2. We then investigated the effect of Cyp4f14 deletion on vitamin E metabolism and status in vivo. Cyp4f14-null mice exhibited substrate-specific reductions in liver microsomal vitamin E-ω-hydroxylase activity ranging from 93% (γ-TOH) to 48% (γ-tocotrienol). In vivo data obtained from metabolic cage studies showed whole-body reductions in metabolism of γ-TOH of 90% and of 68% for δ- and α-TOH. This metabolic deficit in Cyp4f14(-/-) mice was partially offset by increased fecal excretion of nonmetabolized tocopherols and of novel ω-1- and ω-2-hydroxytocopherols. 12'-OH-γ-TOH represented 41% of whole-body production of γ-TOH metabolites in Cyp4f14(-/-) mice fed a soybean oil diet. Despite these counterbalancing mechanisms, Cyp4f14-null mice fed this diet for 6 weeks hyper-accumulated γ-TOH (2-fold increase over wild-type littermates) in all tissues and appeared normal. We conclude that CYP4F14 is the major but not the only vitamin E-ω-hydroxylase in mice. Its disruption significantly impairs whole-body vitamin E metabolism and alters the widely conserved phenotype of preferential tissue deposition of α-TOH. This model animal and its derivatives will be valuable in determining the biological actions of specific tocopherols and tocotrienols in vivo.     

The effect of α-tocopherol, α- and γ-tocotrienols on amyloid-β aggregation and disaggregation in vitro

One of the neuropathological hallmarks of Alzheimer's disease (AD)—causing neurodegeneration and consequent memory deterioration, and eventually, cognitive decline—is amyloid-β (Aβ) aggregation forming amyloid plaques. Our previous study showed the potential of a tocotrienol-rich fraction—a mixture of naturally occurring of vitamin E analogs—to inhibit Aβ aggregation and restore cognitive function in an AD mouse model. The current study examined the effect of three vitamin E analogs—α-tocopherol (α-TOC), α-tocotrienol (α-T3), and γ-tocotrienol (γ-T3)—on Aβ aggregation, disaggregation, and oligomerization in vitro. Thioflavin T (ThT) assay showed α-T3 reduced Aβ aggregation at 10 μM concentration. Furthermore, both α-T3 and γ-T3 demonstrated Aβ disaggregation, as shown by the reduction of ThT fluorescence. However, α-TOC showed no significant effect. We confirmed the results for ThT assays with scanning electron microscopy imaging. Further investigation in photo-induced cross-linking of unmodified protein assay indicated a reduction in Aβ oligomerization by γ-T3. The present study thus revealed the individual effect of each tocotrienol analog in reducing Aβ aggregation and oligomerization as well as disaggregating preformed fibrils.     

Inhibitory effects of γ-tocotrienol on invasion and metastasis of human gastric adenocarcinoma SGC-7901 cells

Natural vitamin E is a mixture of two classes of compounds, tocopherols and tocotrienols. Recent research has revealed that tocotrienols, especially γ-tocotrienol, exhibit not only the same antioxidant ability as tocopherols, but also remarkable anticancer capacity in cancer cell lines. In this study, the invasion and metastatic capacities of gastric adenocarcinoma SGC-7901 cells and the correlation with antimetastasis mechanisms induced by γ-tocotrienol were explored. The results showed the inhibitory effects of γ-tocotrienol at doses of 15, 30, 45 and 60 μmol/L for 48 h on cell migration and cell matrigel invasion; activities of matrix metalloproteinase (MMPs) increased in SGC-7901 cells when compared to the control group (P<.05 or P<.01). An increasing trend in the chemotactic responses to fibronectin (FN) in SGC-7901 cells was found in the γ-tocotrienol treatments. SGC-7901 cell attachment decreased in the γ-tocotrienol-treated groups in comparison with the control group (P<.01). The mRNA expressions of MMP-2 and MMP-9 showed that γ-tocotrienol significantly reduced the matrigel invasion capability through down-regulation of the mRNA expressions of MMP-2 and MMP-9 (P<.01), and up-regulation of tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 in SGC-7901 cells by treatment with γ-tocotrienol for 48 h (P<.05). γ-Tocotrienol also significantly increased the mRNA expression of nm23-H1 in SGC-7901 cells (P<.01). These findings suggest a potential mechanism of γ-tocotrienol-mediated antitumor metastasis activity and indicate the role of vitamin E as potential chemopreventative agents against gastric cancer.     

Concentration of carotenoids, retinol and alpha-tocopherol in plasma of six microchiroptera species

To adequately feed species in captivity it is necessary to know their nutritional habits and their natural availability of specific nutrients. Such essential nutrients are vitamin A, vitamin E and selected carotenoids as vitamin-A-precursors. Because their blood plasma concentration are valid biomarkers of nutritional status of dietary intake, we determined the concentrations of carotenoids, retinol and alpha-tocopherol by HPLC as well as the transport proteins for retinol, the retinol-binding protein (RBP) and transthyretin (TTR) immunologically in the plasma of six species of microchiroptera from free-ranging animals and compared it in one species (Carollia perspicillata) to a group held in captivity. Plasma concentrations of the investigated components were generally much lower compared to most other mammals. Within the bats, differences were observed for all components. As in other species retinol, RBP and TTR were present but no retinyl esters could be detected. Plasma of the insectivorous bat species Molossus molossus contained carotenoids. Within the group of carotenoids, beta-carotene was dominant and only traces of lutein were present. Phyllostomus hastatus revealed the highest alpha-tocopherol concentration. No differences in the plasma content of the investigated compounds were found between a group of Carollia perspicillata kept in captivity for 20 years and free-ranging individuals from a population in Central America. No sex related differences were obvious. In conclusion, nutritional biomarkers in bats were highly variable due to dietary and possible species-specific differences.     

Tissue-Specific antioxidant profiles and susceptibility to lipid peroxidation of the newly hatched chick

The hatching process is characterized by a range of adaptive changes, and a newly hatched chick is considered as an intermediate stage between prenatal and postnatal development. The aim of the present study was to evaluate the characteristic relationships between tissue-specific fatty acid composition and antioxidant protection in newly hatched chicks. Liver, yolk sac membrane, heart, kidney, lung, and four brain regions (cerebrum, cerebellum, stem, and optic lobes) were collected. Fatty acid composition of total lipids and phosphoglycerides, α-tocopherol, lutein, ascorbic acid, reduced glutathione, and the activities of Mn-and Cu,Zn-superoxide dismutase (SOD) and Se-dependent and non-Se-glutathione peroxidase (GSH-Px), and catalase (CAT) were determined. The levels of Fe, Cu, Zn, and Mn as well as tissue susceptibility to lipid peroxidation were also studied. The tissues of the newly hatched chick showed distinctive features in fatty acid profiles, antioxidant accumulation, and susceptibility to lipid peroxidation. The brain clearly displayed the greatest susceptibility to spontaneous and Fe-stimulated lipid peroxidation, was highly unsaturated and contained very low levels of vitamin E, no detectable carotenoids, low GSH-Px, and low CAT activity. At the same time, the brain was characterized by high ascorbic acid concentration and comparatively high SOD activity. It was suggested that in postnatal development, antioxidant enzymes presumably play the major role in antioxidant protection of the chick tissues.     

Retinol and α-tocopherol concentrations in whole fish commonly fed in zoos and aquariums

Retinol (n = 17 spp.) and α-tocopherol (n = 9 spp.) concentrations in whole fish utilized for captive animal feeding programs were determined by high-performance liquid chromatography (HPLC) following routine storage and preparation after commercial purchase by two zoological institutions. Vitamin A activity was calculated from retinol values and ranged from 55 IU/100 g (immature herring) to >2,000 IU/100 g (salmon) on an as-fed basis. α-Tocopherol values, a measure of vitamin E activity, ranged from 0.9 IU/100 g (butterfish) to 12.3 IU/100 g (tilapia) on a wet basis. Vitamin levels in whole fish were intermediate to values previously quantified for muscle or liver tissues alone. Vitamin concentrations in fish livers were quantified separately in seven of these species; liver contributed 35–63% of total retinol measured and 8–34% of total α-tocopherol. Based on these analyses, whole fish commonly fed in zoos, aquariums, and marine zoological parks would appear to meet vitamin A requirements established for most species without additional supplementation, whereas levels of vitamin E quantified indicate a need for supplementation of diets for piscivores.     

The effect of vitamin E analogues and long hydrocarbon chain compounds on calcium-induced muscle damage. A novel role for α-tocopherol?

Previous studies have demonstrated that supplemental α-tocopherol inhibited calcium-induced cytosolic enzyme efflux from normal rat skeletal muscles incubated in vitro and suggested that the protective action was mediated by the phytyl chain of α-tocopherol [1]. In order to investigate this further a number of hydrocarbon chain analogues of tocopherol (7.8-dimethyl tocol, 5,7-dimethyl tocol, tocol, α-tocotrienol, α-tocopherol [10], vitamin K1, vitamin K1 [10], vitamin K1 diacetate, vitamin K2 [20], phytyl ubiquinone and retinol) were tested for any ability to inhibit calcium ionophore, A23187, induced creatine kinase (CK) enzyme efflux. Some compounds were found to be very effective inhibitors and comparison of their structures and ability and to inhibit TBARS production in muscle homogenates revealed that the effects did not appear related to antioxidant capacity or chromanol methyl groups, but rather the length and structure of the hydrocarbon chain was the important mediator of the effects seen.     

γ-Tocopherol abolishes postprandial increases in plasma methylglyoxal following an oral dose of glucose in healthy, college-aged men

Postprandial hyperglycemia contributes to the risk of cardiovascular disease in part by increasing concentrations of the reactive dicarbonyl methylglyoxal (MGO), a byproduct of glucose metabolism. Oxidative stress increases MGO formation from glucose in vitro and decreases its glutathione-dependent detoxification to lactate. We hypothesized that the antioxidant γ-tocopherol, a form of vitamin E, would decrease hyperglycemia-mediated postprandial increases in plasma MGO in healthy, normoglycemic, college-aged men. Participants (n=12 men; 22.3±1.0 years; 29.3±2.4 kg/m(2)) received an oral dose of glucose (75 g) in the fasted state prior to and following 5-day ingestion of a vitamin E supplement enriched in γ-tocopherol (500 mg/day). γ-Tocopherol supplementation increased (P<.0001) plasma γ-tocopherol from 2.22±0.32 to 7.06±0.71 μmol/l. Baseline MGO concentrations and postprandial hyperglycemic responses were unaffected by γ-tocopherol supplementation (P>.05). Postprandial MGO concentrations increased in the absence of supplemental γ-tocopherol (P<.05), but not following γ-tocopherol supplementation (P>.05). Area under the curve for plasma MGO was significantly (P<.05) smaller with the supplementation of γ-tocopherol than without (area under the curve (0-180 min), -778±1010 vs. 2277±705). Plasma concentrations of γ-carboxyethyl-hydroxychroman, reduced glutathione and markers of total antioxidant capacity increased after supplementation, and these markers and plasma γ-tocopherol were inversely correlated with plasma MGO (r=-0.48 to -0.67, P<.05). These data suggest that short-term supplementation of γ-tocopherol abolishes the oral glucose-mediated increases in postprandial MGO through its direct and indirect antioxidant properties and may reduce hyperglycemia-mediated cardiovascular disease risk.