α-Tocopherol reduces VLDL secretion through modulation of intracellular ER-to-Golgi transport of VLDL.

Avoiding hepatic steatosis is crucial for preventing liver dysfunction, and one mechanism by which this is accomplished is through synchronization of the rate of very low density lipoprotein (VLDL) synthesis with its secretion. Endoplasmic reticulum (ER)-to-Golgi transport of nascent VLDL is the rate-limiting step in its secretion and is mediated by the VLDL transport vesicle (VTV). Recent in vivo studies have indicated that α-tocopherol (α-T) supplementation can reverse steatosis in nonalcoholic fatty liver disease, but its effects on hepatic lipoprotein metabolism are poorly understood. Here, we investigated the impact of α-T on hepatic VLDL synthesis, secretion, and intracellular ER-to-Golgi VLDL trafficking using an in vitro model. Pulse-chase assays using [3H]-oleic acid and 100 µmol/L α-T demonstrated a disruption of early VLDL synthesis, resulting in enhanced apolipoprotein B-100 expression, decreased expression in markers for VTV budding, ER-to-Golgi VLDL transport, and reduced VLDL secretion. Additionally, an in vitro VTV budding assay indicated a significant decrease in VTV production and VTV-Golgi fusion. Confocal imaging of lipid droplet (LD) localization revealed a decrease in overall LD retention, diminished presence of ER-associated LDs, and an increase in Golgi-level LD retention. We conclude that α-T disrupts ER-to-Golgi VLDL transport by modulating the expression of specific proteins and thus reduces VLDL secretion.

7ß-Hydroxycholesterol and 7-ketocholesterol: New oxidative stress biomarkers of sarcopenia inducing cytotoxic effects on myoblasts and myotubes.

Aging is a complex biological process which can be associated with skeletal muscle degradation leading to sarcopenia. The aim of this study consisted i) to determine the oxidative and inflammatory status of sarcopenic patients and ii) to clarify the impact of oxidative stress on myoblasts and myotubes. To this end, various biomarkers of inflammation (C-reactive protein (CRP), TNF-α, IL-6, IL-8, leukotriene B4 (LTB4)) and oxidative stress (malondialdehyde, conjugated dienes, carbonylated proteins and antioxidant enzymes: catalase, superoxide dismutase, glutathione peroxidase) as well as oxidized derivatives of cholesterol formed by cholesterol autoxidation (7-ketocholesterol, 7ß-hydroxycholesterol), were analyzed. Apelin, a myokine which contributes to muscle strength, was also quantified. To this end, a case-control study was conducted to evaluate the RedOx and inflammatory status in 45 elderly subjects (23 non-sarcopenic; 22 sarcopenic) from 65 years old and higher. SARCopenia-Formular (SARC-F) and Timed Up and Go (TUG) tests were used to distinguish between sarcopenic and non-sarcopenic subjects. By using red blood cells, plasma and/or serum, we observed in sarcopenic patients an increased activity of major antioxidant enzymes (superoxide dismutase, glutathione peroxidase, catalase) associated with lipid peroxidation and protein carbonylation (increased level of malondialdehyde, conjugated dienes and carbonylated proteins). Higher levels of 7-ketocholesterol and 7ß-hydroxycholesterol were also observed in the plasma of sarcopenic patients. Significant differences were only observed with 7ß-hydroxycholesterol. In sarcopenic patients comparatively to non-sarcopenic subjects, significant increase of CRP, LTB4 and apelin were observed whereas similar levels of TNF-α, IL-6 and IL-8 were found. The increased plasma level of 7-ketocholesterol and 7ß-hydroxycholesterol in sarcopenic patients led us to study the cytotoxic effect of these oxysterols on undifferentiated (myoblasts) and differentiated (myotubes) murine C2C12 cells. With the fluorescein diacetate and sulforhodamine 101 assays, an induction of cell death was observed both on undifferentiated and differentiated cells: the cytotoxic effects were less pronounced with 7-ketocholesterol. In addition, IL-6 secretion was never detected whatever the culture conditions, TNF-α secretion was significantly increased on undifferentiated and differentiated C2C12 cells treated with 7-ketocholesterol- and 7ß-hydroxycholesterol, and IL-8 secretion was increased on differentiated cells. 7-ketocholesterol- and 7ß-hydroxycholesterol-induced cell death was strongly attenuated by α-tocopherol and Pistacia lentiscus L. seed oil both on myoblasts and/or myotubes. TNF-α and/or IL-8 secretions were reduced by α-tocopherol and Pistacia lentiscus L. seed oil. Our data support the hypothesis that the enhancement of oxidative stress observed in sarcopenic patients could contribute, especially via 7ß-hydroxycholesterol, to skeletal muscle atrophy and inflammation via cytotoxic effects on myoblasts and myotubes. These data bring new elements to understand the pathophysiology of sarcopenia and open new perspectives for the treatment of this frequent age-related disease.

Genome-wide scan for oil quality reveals a coregulation mechanism of tocopherols and fatty acids in soybean seeds.

Tocopherols (vitamin E) play essential roles in human health because of their antioxidant activity, and plant-derived oils are the richest sources of tocopherols in the human diet. Although soybean (Glycine max) is one of the main sources of plant-derived oil and tocopherol in the world, the relationship between tocopherol and oil in soybean seeds remains unclear. Here, we focus on dissecting tocopherol metabolism with the long-term goal of increasing α-tocopherol content and soybean oil quality. We first collected tocopherol and fatty acid profiles in a soybean population (>800 soybean accessions) and found that tocopherol content increased during soybean domestication. A strong positive correlation between tocopherol and oil content was also detected. Five tocopherol pathway-related loci were identified using a metabolite genome-wide association study strategy. Genetic variations in three tocopherol pathway genes were responsible for total tocopherol content and composition in the soybean population through effects on enzyme activity, mainly caused by non-conserved amino acid substitution or changes in gene transcription level. Moreover, the fatty acid regulatory transcription factor GmZF351 directly activated tocopherol pathway gene expression, increasing both fatty acid and tocopherol contents in soybean seeds. Our study reveals the functional differentiation of tocopherol pathway genes in soybean populations and provides a framework for development of new soybean varieties with high α-tocopherol content and oil quality in seeds.

The BCO2 Genotype and the Expression of BCO1, BCO2, LRAT, and TTPA Genes in the Adipose Tissue and Brain of Rabbits Fed a Diet with Marigold Flower Extract.

This study was undertaken to evaluate the effect of the BCO2 genotype and dietary supplementation with marigold flower extract on the expression of BCO1, BCO2, LRAT, and TTPA genes in the adipose tissue and brain of rabbits. The concentrations of lutein, zeaxanthin, ß-carotene, retinol, and α-tocopherol were determined in samples collected from rabbits. Sixty young male Termond White rabbits were allocated to three groups based on their genotype at codon 248 of the BCO2 gene (ins/ins, ins/del, and del/del). Each group comprised two subgroups; one subgroup was administered a standard diet, whereas the diet offered to the other subgroup was supplemented with 6 g/kg of marigold flower extract. The study demonstrated that the BCO2 genotype may influence the expression levels of the BCO2, LRAT, and TTPA genes in adipose tissue, and TTPA and BCO1 genes in the brain. Moreover, an increase in the amount of lutein in the diet of BCO2 del/del rabbits may increase the expression of BCO1, LRAT, and TTPA genes in adipose tissue, and the expression of the BCO2 gene in the brain. Another finding of the study is that the content of carotenoids and α-tocopherol increases in both the adipose tissue and brain of BCO2 del/del rabbits.

Antioxidants supplementation improves the quality of in vitro produced ovine embryos with amendments in key development gene expressions.

The present study assessed the effects of supplementation of different antioxidants on oocyte maturation, embryo production, reactive oxygen species (ROS) production and expression of key developmental genes. In this study, using ovine as an animal model, we tested the hypothesis that antioxidant supplementation enhanced the developmental competence of oocytes. Ovine oocytes aspirated from local abattoir-derived ovaries were subjected to IVM with different concentrations of antioxidants [(Melatonin, Ascorbic acid (Vit C), alpha-tocopherol (Vit E), Sodium selenite (SS)]. Oocytes matured without any antioxidant supplementation were used as controls. The oocytes were assessed for maturation rates and ROS levels. Further, embryo production rates in terms of cleavage, blastocysts and total cell numbers were evaluated after performing in vitro fertilization. Real-Time PCR analysis was used to evaluate the expression of stress related gene (SOD-1), growth related (GDF-9, BMP-15), and apoptosis-related genes (BCL-2 and BAX). We observed that maturation rates were significantly higher in alpha-tocopherol (100 µM; 92.4%) groups followed by melatonin (30 µM; 89.1%) group. However, blastocyst rates in ascorbic acid (100 µM; 19.5%), melatonin (30 µM; 18.4%), alpha-tocopherol (100 µM; 18.2%), and sodium selenite (20 µM; 16.9%) groups were significantly higher (P 0.05) than that observed in the control groups. Total cell numbers in blastocysts in the melatonin, ascorbic acid and alpha-tocopherol groups were significantly higher than those observed in sodium selenite and control groups. ROS production was reduced in groups treated with melatonin (30 µM), vitamin C (100 µM), sodium selenite (20 µM) and α-tocopherol (200 µM) compared with that observed in the control group. Supplementation of antioxidants caused the alterations in mRNA expression of growth, stress, and apoptosis related gene expression in matured oocytes. The results recommend that antioxidants alpha-tocopherol (200 µM), sodium selenite (40 µM), melatonin (30 µM) and ascorbic acid (100 µM) during IVM reduced the oxidative stress by decreasing ROS levels in oocytes, thus improving embryo quantity and quality.

Insect cuticular compounds affect Conidiobolus coronatus (Entomopthorales) sporulation and the activity of enzymes involved in fungal infection.

Mycoses are a global problem that affects humans and animals. In the present study, the entomopathogenic soil fungus Conidiobolus coronatus (Entomophthorales), infecting in tropics also humans, sheep and horses, was cultivated with the addition of insect cuticular compounds (CCs) previously detected in the cuticle of C. coronatus-resistant fly species (C10-C30 fatty alcohols, butyl oleate, butyl stearate, glycerol oleate, squalene, tocopherol acetate). Our findings indicate that CCs have diversified and complex effects on the growth and sporulation of C. coronatus and its ability to infect the larvae of Galleria mellonella (Lepidoptera). The CCs affected protein content and cuticle-degrading enzymes (CDEs) activity in the conidia. Some CCs inhibited fungal growth (0.1% C10), decreased sporulation (C12, C16, C24, C28, C30, butyl stearate, squalene), virulence (C12, C14, butyl oleate, butyl stearate) and protein content (C18). They also reduced conidial CDE activity: elastase (C24, butyl oleate, butyl stearate, squalene, tocopherol acetate), chitobiosidase (C12, C14, C20) and lipase (C12, C18, C26, squalene, tocopherol acetate). Several CCs enhanced sporulation (C14, C18, C22, C26, C30), virulence (C18, C26, squalene), conidial protein content (C16, C24, C30, squalene) and CDE activity: elastase (C10, C16, C18), NAGase (C16, C20), chitobiosidase (C16) and lipase (C10, C14, C16, C20, butyl oleate). Our findings indicate that C. coronatus colonies grown on media supplemented with CCs employ various compensation strategies: colonies grown with C16 alcohol demonstrated reduced sporulation but greater conidial protein accumulation and increased elastase, NAGase, chitobiosidase and lipase activity, thus preserving high virulence. Also, colonies supplemented with C18 alcohol demonstrated high virulence and enhanced sporulation and elastase activity but slightly decreased conidial protein content. CCs that inhibit the activity of lipases and proteases show promise in the fight against conidiobolomycosis.

Hepato-Protective Effects of Delta-Tocotrienol and Alpha-Tocopherol in Patients with Non-Alcoholic Fatty Liver Disease: Regulation of Circulating MicroRNA Expression.

MicroRNAs (miRNAs) play a key role in the regulation of genes for normal metabolism in the liver. Dysregulation of miRNAs is involved in the development and progression of non-alcoholic fatty liver disease (NAFLD). We aimed to explore changes in circulating miRNA expression in response to delta-tocotrienol (δT3) and alpha-tocopherol (αTF) supplementation and correlate them with relevant biochemical markers in patients with NAFLD. In total, 100 patients with NAFLD were randomized to either receive δT3 (n = 50) 300 mg or αTF (n = 50) 268 mg twice/day for 48 weeks. Plasma expression of miRNA-122, -21, -103a-2, -421, -375 and -34a were determined at baseline, 24 and 48 weeks of intervention using RT-qPCR. Both δT3 and αTF significantly downregulated expression of miRNA-122, -21, -103a-2, -421, -375 and -34a. Moreover, δT3 was more effective than αTF in reducing expression of miRNA-375 and -34a. A significant correlation was observed between miRNA expression and biochemical markers of hepatic steatosis, insulin resistance (IR), oxidative stress (OS), inflammation and apoptosis. δT3 and αTF exert hepato-protective effects by downregulating miRNAs involved in hepatic steatosis, IR, OS, inflammation and apoptosis in patients with NAFLD. Furthermore, δT3 has more pronounced effects than αTF in reducing miR-375 and miR-34a, which are linked to regulation of inflammation and apoptosis.

The Influence of Rumen-Protected Choline and α-tocopherol Supplementation on Early Lactating Dairy Cows Metabolism.

It is well documented that choline is known as one of the essential ingredients of phospholipids. Choline acts as a determinative element for appropriate cell membrane functions. On the other hand α-tocopherol (Vit E) is a fat-soluble vitamin. This vitamin acts as a strong antioxidant in the living body's defense system against oxidative stress. Lipid peroxidation in peripartum and early lactating cows is significantly increased while the level of serum Vit E is decreases dramatically. These concomitant physiological changes demonstrate a higher level of oxidative stress subsequently leads to serious health issues in dairy cows. Therefore, the present research was designed to investigate the following items in dairy cattle: 1) evaluation of the possible changes in serum protein fractions, and 2) comparing the oxidative status of orally RPC and vitamin E supplementation in dairy cows in early lactation period. In the current study 30 early lactating primiparous and multiparous Holstein cows (body condition score (BCS)=2.51 ± 0.10) were used beginning five weeks postpartum. All the animals were randomly divided in to three groups (n=10) (number of lactation=2.61). The animals were randomly assigned to receive one of the following treatments. Group 1 served as control group were not received any supplement. The second group was supplemented with 90 g/d of RPC (Reashre Choline, Balchem, USA). The third group was administrated 4400 IU/d vitamin E (Roche, Vitamins Ltd; Switzerland). In the current study, serum protein electrophoresis showed four main fractions as follows: albumin, α-globulin, ß-globulin, and γ-globulin. The recorded data showed that the percentages of albumin and γ-globulin fractions were higher in treated groups compared to the control group. In the animals supplementing with RPC and vitamin E the percentages of serum albumin increased to the value of 37. 70±1.63 and 38.21±1.28 respectively compare to the control group (34.69±1.21), which were significant (P<0.05).

Dietary oleuropein extract supplementation and its combination with α-tocopheryl acetate and selenium modifies the free fatty acid profile of pork and improves its stability.

BACKGROUND: Olive-derived antioxidants have been shown to affect the oxidative status of meat and have also been associated with greater consumption of glucose, which might affect glycogen stores and muscle characteristics. This study evaluated the effect of oleuropein extract supplementation (OLE) versus vitamin E + Se (VE), and their combination (VEOLE), in pig diets, on pH, drip loss, the proportion of free fatty acids, and meat stability, and their prediction by blood oxidative status markers. RESULTS: The drip loss of muscle was lower in antioxidant-supplemented groups when compared with controls. α-Tocopherol concentration and total fatty acids profile were not affected by dietary oleuropein supplementation. However, OLE and VEOLE had lower free n-3 polyunsaturated fatty acid (PUFA) levels when compared with VE and tended to have higher free monounsaturated fatty acid (MUFA) levels. Furthermore, the VEOLE group had lower free n-6 PUFA levels when compared with controls or VE, whereas the OLE group had intermediated values. Muscle samples from pigs subjected to the antioxidant-mixed supplementation (VEOLE) had lower malondialdehyde concentration when compared with the others. The VE and OLE groups showed intermediate malondialdehyde values. Chilled meat stability was highly correlated with antioxidant status in vivo. CONCLUSION: The administration of 96 mg oleuropein kg-1 feed produced similar meat quality characteristics as the use of 100 mg kg-1 α-tocopheryl acetate +0.26 mg kg-1 sodium selenite and it would be an interesting alternative in Mediterranean countries. The VEOLE group was the most effective for reducing lipid oxidation and for the production of polyunsaturated free fatty acids in meat, which would result in lower rancidity formation and better aroma development in products. © 2020 Society of Chemical Industry.

Impact of alpha-tocopherol deficiency and supplementation on sacrocaudalis and gluteal muscle fiber histopathology and morphology in horses.

BACKGROUND: A subset of horses deficient in alpha-tocopherol (α-TP) develop muscle atrophy and vitamin E-responsive myopathy (VEM) characterized by mitochondrial alterations in the sacrocaudalis dorsalis medialis muscle (SC). OBJECTIVES: To quantify muscle histopathologic abnormalities in subclinical α-TP deficient horses before and after α-TP supplementation and compare with retrospective (r)VEM cases. ANIMALS: Prospective study; 16 healthy α-TP-deficient Quarter Horses. Retrospective study; 10 retrospective vitamin E-responsive myopathy (rVEM) cases . METHODS: Blood, SC, and gluteus medius (GM) biopsy specimens were obtained before (day 0) and 56 days after 5000 IU/450 kg horse/day PO water dispersible liquid α-TP (n = 8) or control (n = 8). Muscle fiber morphology and mitochondrial alterations were compared in samples from days 0 and 56 and in rVEM cases. RESULTS: Mitochondrial alterations more common than our reference range (<2.5% affected fibers) were present in 3/8 control and 4/8 treatment horses on day 0 in SC but not in GM (mean, 2.2; range, 0%-10% of fibers). Supplementation with α-TP for 56 days did not change the percentage of fibers with mitochondrial alterations or anguloid atrophy, or fiber size in GM or SC. Clinical rVEM horses had significantly more mitochondrial alterations (rVEM SC, 13% ± 7%; GM, 3% ± 2%) and anguloid atrophy compared to subclinical day 0 horses. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinically normal α-TP-deficient horses can have mitochondrial alterations in the SC that are less severe than in atrophied VEM cases and do not resolve after 56 days of α-TP supplementation. Preventing α-TP deficiency may be of long-term importance for mitochondrial viability.

Effect of adding alpha-tocopherol on the oxidation advance during in vitro gastrointestinal digestion of sunflower and flaxseed oils.

Few in vitro studies have tackled the effect of alpha-tocopherol on lipid oxidation during digestion, and discrepant results have been reported. As a result, the aim of this study was to elucidate whether the addition of alpha-tocopherol enhances or slows down the advance of oxidation that occurs during in vitro gastrointestinal digestion of polyunsaturated lipids. For this purpose, commercial sunflower and flaxseed oils (as models of omega-6 and omega-3 rich lipid systems, respectively) were in vitro digested in the absence or in the presence of this tocol at different concentrations (0.02%, 0.2% and 2%). Proton Nuclear Magnetic Resonance (1H NMR) and Solid Phase Microextraction followed by Gas Chromatography/Mass Spectrometry (SPME-GC/MS) were used to investigate in detail potential differences among the digests regarding lipolysis and oxidation level. Alpha-tocopherol addition did not affect the advance of lipolysis, whereas lipid oxidation was enhanced in a dose-dependent manner. In this regard, the increased degradation of polyunsaturated lipids and greater generation of primary and secondary oxidation products observed at higher concentrations of alpha-tocopherol confirmed this observation. Among the formed oxidation products, hydroperoxy-, hydroxy- and keto-dienes, as well as oxygenated alpha,beta-unsaturated aldehydes are worth mentioning. The in vitro bioaccessibility of added tocopherol was estimated to be very low, suggesting a notable transformation under the assayed conditions. Further in vivo studies are necessary to confirm this prooxidant activity of alpha-tocopherol during gastrointestinal digestion.

Adding α-tocopherol-selenium and ascorbic acid to periparturient sow diets influences hemogram, lipid profile, leptin, oxidant/antioxidant imbalance, performance and neonatal piglet mortality.

Alpha-tocopherol-selenium (ATS) and ascorbic acid (AA) are the potent antioxidants. The present study investigated whether supplementation of ATS and AA in periparturient sows has positive effects on amelioration of oxidative stress, serum immunoglobulin G (IgG), lipid profile and sows performance. For this, twenty-four pregnant multiparous sows (landrace×indigenous) were randomly distributed into four groups (6 sows per group) 20 days before expected date of farrowing as Control (basal diet); ATS (basal diet + ATS); AA (basal diet + AA) and ATS-AA (basal diet + ATS plus AA). The results of the study revealed that the concentrations of triglyceride and cholesterol significantly reduced from day -7 to day 7 of farrowing irrespective of supplementations to sows, but the leptin concentration significantly reduced on day 7 of farrowing in ATS-AA supplemented sows (p<0.05). Moreover, sows of supplemented groups experienced decreased oxidative stress and cortisol level than control sows. The serum IgG concentration was significantly increased on day 7 post-farrowing in ATS group but it was much earlier on day 2 of farrowing in ATS-AA group (p<0.001). Supplementing sows with ATS and/or AA did not influence significantly the birth weight, weaning weight and litter size at weaning (p>0.05). Although piglet survival rate was not affected significantly by supplementation, however, piglet mortality rate was lowest in ATS-AA than any other groups. It was concluded that supplementation of ATS and/or AA to sows during late gestating and early lactating period ameliorated oxidative stress, improved lipid profile and serum IgG level without influencing reproductive performance.

The effects of hesperidin or naringin dietary supplementation on the milk properties of dairy ewes.

BACKGROUND: Fortification of animal products with natural bioactive compounds appears to improve their quality and protects consumers against oxidation effects. An experiment was therefore conducted to examine the effects of hesperidin or naringin on milk oxidative stability, yield, composition, coagulation properties, and the fatty acid profile in dairy sheep. Thirty-six Chios ewes were assigned to four groups. A control group was fed a concentrate diet without supplementation. The other three groups were provided with the same diet further supplemented with hesperidin (6000 mg kg-1 ), naringin (6000 mg kg-1 ), or α-tocopheryl acetate (200 mg kg-1 ). The efficacy of flavonoids after a change in diet composition that lowered milk oxidation values by itself was also tested. The duration of the experiment was 28 days with alfalfa hay being the only forage source for the first 14 days, whereas after the 15th day a mixture of alfalfa hay and wheat straw (65:35) was provided. RESULTS: The oxidative stability of milk was improved after 14 days of addition of the examined flavonoids (P < 0.05). Milk malondialdehyde (MDA) values were also decreased as a result of flavonoid dietary supplementation, 14 days after the modification of the forage source. On the other hand, no significant differences in yield, chemical composition, coagulation properties, and fatty acid profile of ewe milk were observed among the treatments throughout the experiment. CONCLUSION: Enrichment of dairy ewes' diets with hesperidin and naringin might be effective in improving milk's oxidative stability without any effects on the milk's chemical composition, coagulation properties, and fatty acid profile. © 2019 Society of Chemical Industry.

Activation of human insulin by vitamin E: A molecular dynamics simulation study.

Lack of perfect insulin signaling can lead to the insulin resistance, which is the hallmark of diabetes mellitus. Activation of insulin and its binding to the receptor for signaling process initiates via B-chain C-terminal hinge conformational change through an open structure to "wide-open" conformation. Observational studies and basic scientific evidence suggest that vitamin D and E directly and/or indirectly prevent diabetes through improving glucose secretion and tolerance, activating calcium dependent endopeptidases and thus improving insulin exocytosis, antioxidant effect and reducing insulin resistance. On the contrary, clinical trials have yielded inconsistent results about the efficacy of vitamin D supplementations for the control of glucose hemostasis. In this work, best binding modes of vitamin D3 and E on insulin obtained from AutoDock Vina were selected for Molecular Dynamic, MD, study. The binding energy obtained from Molecular Mechanics- Poisson Boltzman Surface Area, MM-PBSA method, revealed that Vitamins D3 and E have good affinity to bind to the insulin and vitamin E has higher binding energy (-46 kj/mol) by engaging more residues in binding site. Distance and angle calculation results illustrated that vitamin E changes the B-chain conformation and it causes the formation of wide-open/active form of insulin. Vitamin E increases the ValB12-TyrB26 distance to ∼15 Šand changes the hinge angle to ∼65°. Consequently, essential hydrophobic residues for binding to insulin receptor exposed to surface in the presence of vitamin E. However, our data illustrated that vitamin D3 cannot change B-chain conformation. Thus our MD simulations propose a model for insulin activation through vitamin E interaction for therapeutic approaches.

Effect of vitamin E sources and inclusion levels in diets on growth performance, meat quality, alpha-tocopherol retention, and intestinal inflammatory cytokine expression in broiler chickens.

Water-miscible vitamin E (WVE) has been known to be more bioavailable than conventional lipid-soluble vitamin E (LVE) in human foods because of its greater solubility in the gastrointestinal tract. However, no data regarding the comparison of the efficacy of WVE to LVE were available in poultry diets. The objective of the current experiment was to investigate the effect of vitamin E (VE) sources and inclusion levels in diets on growth performance, meat quality, alpha-tocopherol retention, and intestinal inflammatory cytokine expression in broiler chickens. A total of 420 6-d-old broiler chicks were allotted to 1 of 7 dietary treatments with 6 replicates in a completely randomized design. The corn-soybean meal-based basal diet was formulated without inclusion of supplemental VE. Additional 6 diets were prepared by supplementing the basal diet with either WVE or LVE at the inclusion levels of 33, 65, or 100 IU/kg in diets. Diets were fed to birds for 26 d. Results indicated that different VE sources and inclusion levels from 33 to 100 IU/kg in diets had no effects on growth performance in broilers. The VE sources did not affect lipid oxidation in both breast and thigh meat. However, increasing inclusion levels of VE decreased (linear and quadratic, P < 0.05) lipid oxidation in breast meat. Regardless of VE sources, increasing inclusion levels of VE increased (linear, P < 0.01) alpha-tocopherol concentrations in breast meat. Dietary WVE showed a greater reduction in the expression of both pro- and anti-inflammatory cytokine genes in the jejunum than dietary LVE. Increasing inclusion levels of both VE sources decreased (linear and quadratic, P < 0.05) the expression of both pro- and anti-inflammatory cytokine genes in the jejunum. The efficacy of dietary WVE for growth performance, meat quality, lipid oxidation, and alpha-tocopherol retention in broilers chickens is similar to that of dietary LVE. However, it is likely that dietary WVE is more effective in decreasing intestinal inflammatory responses than dietary LVE in broilers.

ROS management is mediated by ascorbate-glutathione-α-tocopherol triad in co-ordination with secondary metabolic pathway under cadmium stress in Withania somnifera.

Being static, plants are frequently exposed to various essential and non-essential heavy metals from the surroundings. This exposure results in considerable ROS generation leading to oxidative stress, the primary response of the plants under heavy metal stress. Withania somnifera is a reputed Indian medicinal plant in Ayurveda, having various pharmacological activities due to the presence of withanolides. The present study deals with the understanding endurance of oxidative stress caused by heavy metal exposure and its management through antioxidant partners in synchronization with secondary metabolites in W. somnifera. The quantitative assessment of enzymatic/non-enzymatic antioxidants revealed significant participation of ascorbate-glutathione-α-tocopherol triad in ROS management. Higher activities of glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) resulted in glutathione and ascorbate accumulation. In addition, superoxide dismutase (SOD), glutathione peroxidase (GPX) and peroxidase (POD) were contributed considerably in ROS homeostasis maintenance. In-situ localization and assays related to ROS generation/scavenging revealed key management of ROS status under Cd stress. Higher antioxidative and reducing power activity attributed to the tolerance capability to the plant. Increased expression of withanolide biosynthetic pathway genes such as WsHMGR, WsDXS, WsDXR and WsCAS correlated with enhanced withanolides. The present study indicated the crucial role of the ascorbate-glutathione-α-tocopherol triad in co-ordination with withanolide biosynthesis in affording the oxidative stress, possibly through a cross-talk between the antioxidant machinery and secondary metabolite biosynthesis. The knowledge may be useful in providing the guidelines for developing abiotic stress resistance in plants using conventional and molecular approaches.

Effect of milking frequency and α-tocopherol plus selenium supplementation on sheep milk lipid composition and oxidative stability.

The aim of this research was to study the effect of milking frequency [once-daily milking (ODM) vs. twice-daily milking (TDM)] and antioxidant (AOX) supplementation on fatty acid (FA) profile and oxidative stability in sheep milk. Sixteen Assaf ewes were used; 8 did not receive any vitamin-mineral supplement (control), and the other 8 received an oral dose of 1,000 IU of α-tocopherol and 0.4 mg of Se daily. The experiment consisted of 2 consecutive periods; the first was 3 wk with TDM of both mammary glands. The second period was 8 wk and consisted of ODM of one mammary gland and TDM of the other gland. All ewes were fed ad libitum the same total mixed ration from lambing and throughout the experiment. There were no differences in plasma or milk Se concentrations between control and AOX ewes. However, plasma and milk α-tocopherol concentrations and AOX capacity were increased in ewes receiving the AOX supplement. Milk FA profile was practically unaffected after 21 d of AOX supplementation. However, after 77 d, AOX supplementation increased the relative percentage of C16:0 and cis-9 C18:1 and reduced the proportions of some saturated FA with less than 16 carbons and cis-9 C12:1. Antioxidant supplementation had no effect on the proportions of conjugated linoleic acid or total polyunsaturated FA (PUFA) but decreased the proportion of trans-7,cis-9 C18:2 and increased that of n-6 C20:3. Once-daily milking did not affect α-tocopherol, Se, or fat resistance to oxidation in milk. Total monounsaturated FA, cis-9 C16:1, and several cis and trans isomers of C18:1 were increased and total saturated FA were decreased in milk from ODM glands. Compared with TDM, ODM increased the proportions of cis-9,cis-12 C18:2 and several isomers of C18:2 and reduced those of cis-9,cis-12,cis-15 C18:3 and some PUFA of 20 and 22 carbons, but total proportion of PUFA was unaffected. Once-daily milking and AOX supplementation modified milk FA profile, but the effects of ODM could be considered of little biological relevance for consumer health. Supplementing ewes with α-tocopherol plus Se could be considered an effective strategy to improve plasma AOX status and reduce milk fat oxidation without substantial changes in the milk FA profile.

Identification and expression analysis of alpha tocopherol transfer protein in chickens fed diets containing different concentrations of alpha-tocopherol.

Among the eight forms of vitamin E, the liver preferentially releases α-tocopherol into the circulation and it is distributed to the non-liver tissues. In the hepatocytes, alpha tocopherol transfer protein (TTPA) specifically recognizes α-tocopherol with 2R-configuration and facilitates its intracellular transfer. The identification and characterization of TTPA expression have not been demonstrated in avian species. Therefore, the objectives of this study were to identify avian TTPAs, to compare the sequence conservation, phylogenetic relationship, protein interactions, and disease associations of chicken TTPA with those of human and vertebrate TTPA, and to characterize the tissue expression of the TTPA gene in chickens fed diets supplemented with different amounts of α-tocopherol. Our results suggest that the chicken TTPA was highly conserved with the human and vertebrate TTPA, and consisted of a cellular retinaldehyde binding protein and TRIO guanine exchange factor (CRAL_TRIO) domain. Feeding diets supplemented with increasing amounts of α-tocopherol (25 IU/Kg, 50 IU/Kg, or 100 IU/Kg) to broiler chickens had no effects on growth performance compared with feeding basal diets containing no supplemental α-tocopherol. The expression of TTPA gene was detected high in the liver of chickens in response to dietary α-tocopherol concentrations, whereas its expression was very low or undetectable in the non-liver tissues. In conclusion, the chicken TTPA protein sequence is highly conserved with other avian and vertebrate TTPA protein sequences. The higher expression of TTPA gene in the chicken liver in response to dietary α-tocopherol concentrations may suggest its crucial role in transporting α-tocopherol in the chicken liver.

Antioxidant status, meat oxidative stability and quality characteristics of lambs fed with hesperidin, naringin or α-tocopheryl acetate supplemented diets.

BACKGROUND: Dietary supplementation of sheep with bioactive compounds such as citrus flavonoids appears as a great alternative for improving meat antioxidant capacity and could also have beneficial effects on growth performance, carcass traits and meat quality characteristics. An experiment was therefore conducted to examine the effects of hesperidin or naringin dietary supplementation on lambs' growth performance, antioxidant status and meat quality characteristics. Forty-four male 3-month-old Chios lambs were randomly assigned to four groups: C, fed with a basal diet, or H or N or VE offered the same diet further supplemented with hesperidin at 2500 mg or naringin at 2500 mg or α-tocopheryl acetate (vitamin E) at 200 mg kg-1 feed respectively. At the end of the experiment (day 35), samples of longissimus thoracis muscle were collected for meat quality evaluation. RESULTS: Both flavonoids and vitamin E dietary supplementation reduced plasma malondialdehyde (MDA) levels on day 35 (P < 0.05). No significant differences were observed in performance traits and meat quality characteristics among the experimental groups (P > 0.05). Hesperidin and naringin also reduced MDA levels in meat stored at 4 °C for up to 8 days (P < 0.05), although to a lesser extent than α-tocopheryl acetate. CONCLUSION: Incorporation of flavonoids in sheep diets resulted in an improvement of plasma and meat antioxidant capacity. © 2018 Society of Chemical Industry.

Coffee, but Neither Decaffeinated Coffee nor Caffeine, Elicits Chemoprotection Against a Direct Carcinogen in the Colon of Wistar Rats.

Colorectal cancer (CRC) is the third most frequent malignancy worldwide. Coffee is the second most consumed drink in the globe and suggested to decrease the CRC risk. Here, we explored whether coffee, decaffeinated coffee, or caffeine impact on the development of colorectal carcinogenesis induced by the direct carcinogen N-methyl-N-nitro-N-nitrosoguanidine (MNNG) in rats. To this end, sixty-four young male Wistar rats were divided into eight groups of eight animals each. We analyzed the frequency of dysplastic crypts and expression of metallothionein as a biomarker of the cancer risk, as well the expression of phosphorylated H2A histone family/member X (γH2AX) for DNA damage and cyclooxygenase-2 (COX-2) for inflammatory response. We also studied the oxidative stress profile in hepatic and colonic frozen samples (malondialdehyde [MDA], glutathione [GSH], and α-tocopherol). We found that coffee but neither decaffeinated coffee nor caffeine decreased the development of dysplastic crypts in MNNG-exposed rats. All treatments reduced DNA damage intensity in colonocytes. Only decaffeinated coffee increased the numbers of metallothionein positive crypts in comparison with coffee-treated rats. Coffee and caffeine inhibited COX-2 expression in the colon. Both decaffeinated coffee and caffeine decreased hepatic α-tocopherol levels. We suggest that coffee may have other compounds that elicit greater chemoprotective effects than caffeine reducing the CRC risk.