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Effect of supranutritional level of dietary alpha-tocopheryl acetate and selenium on rabbit semen.

This research examined the effects of dietary alpha-tocopheryl acetate (50 or 200 mg/kg diet) and selenium (Se, 0 or 0.5 ppm) supplementation on motion characteristics, oxidative stability and fertilizing ability of rabbit spermatozoa, fresh and stored for 24 h at 5 degrees C. The higher amount of dietary alpha-tocopheryl acetate increased the level of Vitamin E in the fresh semen (1.75 mmol/l versus 0.95 mmol/l) and its oxidative stability (thiobarbituric acid reactive substances-TBARS 12.44 nmol malondialdehyde/10(8) sperm versus 21.4 nmol malondialdehyde/10(8) sperm). Dietary Se increased gluthatione peroxidase activity (GPx) in erythrocytes (285 U/g Hb versus 207 U/g Hb), seminal plasma (270 U/l versus 190 U/l) and spermatozoa (1338 mU/10(9) sperm versus 1103 mU/10(9) sperm), whereas it did not show any effect on alpha-tocopherol level and TBARS. No synergy between Vitamin E and Se was shown. Storage for 24 h at 5 degrees C increased the TBARS level in all the experimental groups. Neither live and acrosome reacted spermatozoa, nor kinetic parameters, nor fertility rate were modified by dietary supplementation.

Animal Nutritional Physiological Phenomena↗

Estradiol enhances the resistance of LDL to oxidation by stabilizing apoB-100 conformation.

Among different proposed mechanisms to account for the protection exerted by estrogens against cardiovascular diseases, the antioxidant effect has attracted considerable attention. We confirmed that 17-beta-estradiol (E2), when added to human LDL at a 6:1 ratio to apoB-100, markedly delays the phase of massive LDL lipid peroxidation induced by Cu(2+). We also observed an increased oxidative resistance of E2-treated LDL by monitoring the early phase of oxidative degradation on the basis of increased LDL surface polarity by the generalized polarization of the lipophilic fluorescent probe 2-(dimethylamino)-6-lauroylnaphthalene (Laurdan). A scavenging of free radicals by E2 is ruled out since, consistent with its structure, its rate constant for the reduction of peroxy radicals is extremely low, i.e., 0.02% of that of vitamin E. Tryptophan fluorescence lifetime and circular dichroism measurements revealed that (i) apoB-100 undergoes a conformational modification and a progressive loss of secondary structure during lipid peroxidation; (ii) E2 increases apoB-100 secondary structure and modifies its conformation; and (iii) the apoB-100 conformational change induced by E2 makes this protein resistant to modifications brought about by lipid peroxidation. We propose that E2, by affecting apoB-100 secondary structure and conformation, modifies the interaction of this protein with the outer layer of the LDL particle thus increasing its overall oxidative resistance.

2-Naphthylamine↗

Antioxidant activity of dietary oregano essential oil and alpha-tocopheryl acetate supplementation in long-term frozen stored turkey meat.

The effects of dietary oregano essential oil and alpha-tocopheryl acetate supplementation on the oxidative stability of long-term frozen stored turkey meat were investigated. Thirty 12-week-old turkeys, randomly divided into five groups, were given a basal diet or a basal diet supplemented with 200 mg of alpha-tocopheryl acetate kg(-1), or 100 or 200 mg of oregano oil kg(-1), or 100 mg of oregano oil plus 100 mg of alpha-tocopheryl acetate kg(-1) for 4 weeks prior to slaughter. Lipid oxidation in breast and thigh meat was assessed after 1, 3, 6, and 9 months of frozen storage at -20 degrees C prior to or following 7 days of refrigerated storage at 4 degrees C. Results showed that oregano oil increased the oxidative stability of breast and thigh meat during the frozen storage. Dietary oregano oil at the inclusion level of 200 mg kg(-1) feed was significantly (p < 0.05) more effective in delaying lipid oxidation compared to the level of 100 mg kg(-1), but equivalent to dietary alpha-tocopheryl acetate supplementation at 200 mg kg(-1), which in turn was inferior to dietary supplementation of 100 mg kg(-1) oregano essential oil plus 100 mg kg(-1) alpha-tocopheryl acetate that was significantly (p < 0.05) superior to all other treatments. Thigh meat was more susceptible to oxidation than breast meat, although the former contained alpha-tocopherol at markedly higher levels. Mean alpha-tocopherol levels in breast and thigh meat from all treatments decreased during the frozen storage, the decrease being sharper between 1 and 3 months of frozen storage for breast and between 3 and 6 months for thigh meat. Oregano oil supplementation increased (p < 0.05) the retention of alpha-tocopherol in meat, the increase being positively correlated with the supplementation level. However, the retention of alpha-tocopherol in meat could only partly elucidate the antioxidant activity exhibited by dietary oregano oil supplementation.

Animals↗

Evidence that inhibitory factor extracted from bovine retractor penis is nitrite, whose acid-activated derivative is stabilized nitric oxide.

1. Unactivated extracts of bovine retractor penis (BRP) contains 3-7 microM nitrite. Acid-activation of these extracts at pH 2 for 10 min followed by neutralization generates the active form of inhibitory factor (IF; assayed by its vasodilator action on rabbit aorta), and is associated with partial loss of nitrite. 2. Increasing the time of acid-activation at pH 2 from 10 to 60 min with intermittent vortex mixing generates greater vasodilator activity and increases nitrite loss. 3. When acid-activated and neutralized extracts are incubated at 37 degrees C or 30 min or boiled for 5 min, vasodilator activity is lost and nitrite content increased. Reactivation of these samples at pH 2 for 10 min followed by neutralization leads to partial recoveries of vasodilator activity with loss in nitrite content. 4. Addition of sodium nitrite to BRP extracts increases acid-activatable vasodilator activity pro rata. 5. Acid-activation of aqueous sodium nitrite solutions results in less loss of nitrite and generation of less vasodilator activity than BRP extracts. Vasodilatation is only transient and is rapidly abolished on neutralization, whereas responses to acid-activated BRP extracts are more prolonged and activity is stable on ice. 6. Bovine aortic endothelial cells yield vasodilator activity that is indistinguishable from that isolated from BRP. It is activated by acid, stable on ice, abolished by boiling or by haemoglobin, and appears to be due to the generation of nitric oxide (NO) from nitrite. 7. The data provide confirmatory evidence that nitrite in BRP extracts is IF, that acid-activation of BRP extracts yields NO which is responsible for its vasodilator action, and that inactivation occurs by decay of NO to nitrite and nitrate. They further suggest that BRP extracts contain a NO-stabilizing agent which favours conversion of nitrite to NO. 8. The finding that bovine aortic endothelial cells yield an agent indistinguishable from IF suggests that nitrite in endothelial cells may likewise be the precursor of endothelium-derived relaxing factor (EDRF), itself identified as NO.

Animals↗

Preparation and characterization of superparamagnetic iron oxide nanoparticles stabilized by alginate.

SPION with appropriate surface chemistry have been widely used experimentally for numerous in vivo applications. In this study, SPION stabilized by alginate (SPION-alginate) were prepared by a modified coprecipitation method. The structure, size, morphology, magnetic property and relaxivity of the SPION-alginate were characterized systematically by means of XRD, TEM, ESEM, AFM, DLS, SQUID magnetometer and MRI, respectively, and the interaction between alginate and iron oxide (Fe(3)O(4)) was characterized by FT-IR and AFM. The results revealed that typical iron oxide nanoparticles were Fe(3)O(4) with a core diameter of 5-10 nm and SPION-alginate had a hydrodynamic diameter of 193.8-483.2 nm. From the magnetization curve, the Ms of a suspension of SPION-alginate was 40 emu/g, corresponding to 73% of that of solid SPION-alginate. This high Ms may be due to the binding of Fe(3)O(4) nanoparticles to alginate macromolecule strands as visually confirmed by AFM. SPION-alginate of several hundred nanometers was stable in size for 12 months at 4 degrees C. Moreover, T1 relaxivity and T2 relaxivity of SPION-alginate in saline (1.5 T, 20 degrees C) were 7.86+/-0.20 s(-1) mM(-1) and 281.2+/-26.4 s(-1) mM(-1), respectively.

Alginates↗

Stability of oxidized Escherichia coli thioredoxin and its dependence on protonation of the aspartic acid residue in the 26 position.

The effects of pH in the range 6.0-8.0 on the thermodynamics of the reversible thermal unfolding of Escherichia coli thioredoxin in the oxidized state have been determined over a range of concentrations using differential scanning calorimetry. The thermal denaturation indicated an inverse temperature dependence on concentration. The data were shown to fit a model based on dimerization of both the native and denatured states of the protein. The degree of dimerization of both states was found to be pH dependent. The previously described importance of protonation of the anomalously titrating aspartic acid 26 residue [Langsetmo, K., Fuchs, J., & Woodward, C. (1991) Biochemistry 30 ,7603-7609] was apparently verified by the agreement between the experimentally determined delta delta Gzerod and the calculated delta delta GzeroH in the pH range 7.0-8.0.

Aspartic Acid↗

[The redox status of the blood plasma in cattle from ecological farming--1. Influence of genotype and age on the antioxidant defence capacity in the blood plasma].

The present study examined the effects of race and age on the redox potential behavior of blood plasma samples. The oxidative stability was studied in the reaction with p-benzoquinone. The investigated animals represented three different cattle races (Limousin, Angus and Hereford), grazed on pasture. The blood samples from clinical healthy heifers (44) and calves (45) were investigated. The following parameters were determined: the start level of redox potential and the level after adding 10, 20, and 30 mg p-benzoquinone to the samples. The analyses of the redox potentials showed the advantage of oxidative stability of heifers compared with calves. The results showed a significant effect of race on the redox potential behavior of blood plasma.

Age Factors↗

Influence of heated vegetable oils and alpha-tocopheryl acetate supplementation on alpha-tocopherol, fatty acids and lipid peroxidation in chicken muscle.

1. Chicks were fed on diets containing fresh, heated or alpha-tocopheryl acetate-supplemented heated vegetable oils. The effects on alpha-tocopherol status, and on the fatty acid composition and oxidative stability of thigh and breast muscle were determined. 2. Plasma alpha-tocopherol was significantly correlated with alpha-tocopherol concentrations in thigh and breast muscle. 3. The fatty acid profiles of muscle lipids reflected dietary fatty acid composition. 4. The consumption of heated sunflower and linseed oils reduced alpha-tocopherol status, altered fatty acid composition of muscle lipids and increased susceptibility of muscle to lipid oxidation. 5. Supplementation of diets containing heated oils with alpha-tocopheryl acetate resulted in some alleviation of these effects. 6. The results indicate that caution should be exercised in the use of thermally oxidised oils in poultry diets if undesirable changes in composition and oxidative stability of carcase lipids are to be avoided.

Animals↗

Enhancing stability and oxidation activity of cytochrome C by immobilization in the nanochannels of mesoporous aluminosilicates.

Hydrothermally stable and structrurally ordered mesoporous and microporous aluminosilicates with different pore sizes have been synthesized to immobilize cytochrome c (cyt c): MAS-9 (pore size 90 A), MCM-48-S (27 A), MCM-41-S (25 A), and Y zeolites (7.4 A). The amount of cyt c adsorption could be increased by the introduction of aluminum into the framework of pure silica materials. Among these mesoprous silicas (MPS), MAS-9 showed the highest loading capacity due to its large pore size. However, cyt c immobilized in MAS-9 could undergo facile unfolding during hydrothermal treatments. MCM-41-S and MCM-48-S have the pore sizes that match well the size of cyt c (25 x 25 x 37 A). Hence the adsorbed cyt c in these two medium pore size MPS have the highest hydrothermal stability and overall catalytic activity. On the other hand, the pore size of NaY zeolite is so small that cyt c is mostly adsorbed only on the outer surface and loses its enzymatic activity rapidly. The improved stability and high catalytic activity of cyt c immobilized in MPS are attributed to the electrostatic attraction between the pore surface and cyt c and the confinement provided by nanochannels. We further observed that cyt c immobilized in MPS exists in both high and low spin states, as inferred from the ESR and UV-vis studies. This is different from the native cyt c, which shows primarily the low spin state. The high spin state arises from the replacement of Met-80 ligands of heme Fe (III) by water or silanol group on silica surface, which could open up the heme groove for easy access of oxidants and substrates to iron center and facilitate the catalytic activity. In the catalytic study, MAS-9-cyt c showed the highest specific activity toward the oxidation of polycyclic aromatic hydrocarbons (PAHs), which arises from the fast mass transfer rate of reaction substrate due to its large pore size. For pinacyanol (a hydrophilic substrate), MCM-41-S-cyt c and MCM-48-S-cyt c showed higher specific activity than NaY-cyt c and MAS-9-cyt c. The result indicated that cyt c embedded in the channels of MCM-41-S and MCM-48-S was protected against unfolding and loss of activity. By increasing the concentration of the spin trapping agent, 5,5-dimethyl-1-pyrroline N-oxide (DMPO) in ESR experiments, we showed that cyt c catalyzes a homolytic cleavage of the O-O bond of hydroperoxide and generates a protein cation radical (g = 2.00). Possible mechanisms for MPS-cyt c catalytic oxidation of hydroperoxides and PAHs are proposed based on the spectroscopic characterizations of the systems.

Aluminum Silicates↗

Effects of dietary vitamin E and fat supplementation on pork quality.

The effects of dietary vitamin E (VE, alpha-tocopherol acetate) and fat supplementation on growth and carcass quality characteristics, oxidative stability of fresh and cooked pork patty in storage, fatty acid profiles of muscle and adipose tissue, and VE concentrations of plasma, muscle, and adipose tissue were studied. Six hundred pigs were allocated to 1 of 6 diets and fed for 63 d in a 3 x 2 factorial design. The dietary treatments included 3 fat levels (normal corn, high oil corn, high oil corn plus added beef tallow) and 2 levels of VE supplementation (40 IU/kg, normal VE supplementation; and 200 IU/kg, high VE supplementation). At 113 kg of BW, 54 pigs were slaughtered as a subsample to evaluate dietary effects on pork quality. Growth performance and meat quality characteristics did not differ (P > 0.05) among treatment groups. The high level of VE supplementation had a beneficial effect on the oxidative stability of pork as indicated by thiobarbituric acid reactive substance (TBARS) values. Lean tissue had lower (P < 0.05) TBARS in the group fed the high VE than in those fed the normal VE level. The TBARS values differed among storage periods (0 to 6 d) and also between fresh and cooked ground ham. Fat type did not significantly affect total saturated and unsaturated fatty acids proportions in the neutral and polar fraction of muscle. Adding VE acetate led to greater (P < 0.05) monounsaturated and total unsaturated fatty acid proportions in neutral lipids of muscle and adipose tissues. Increasing dietary levels of VE acetate increased the concentration of VE in plasma and muscle. These results indicate that dietary VE acetate supplementation increased (P < 0.05) lipid stability and the VE concentration of muscle.

Animal Feed↗

Incorporation of alpha-tocopherol in marine lipid-based liposomes: in vitro and in vivo studies.

Liposomes made from a natural marine lipid extract and containing a high polyunsaturated n-3 fatty lipid ratio were envisaged as oral route vectors and a potential alpha-tocopherol supplement. The behavior of vesicles obtained by simple filtration and of giant vesicles prepared by electroformation was investigated in gastrointestinal-like conditions. The influence of alpha-tocopherol incorporation into liposomes was studied on both physical and chemical membrane stability. Propanal, as an oxidation product of n-3 polyunsaturated fatty acids, was quantified by static headspace gas chromatography when alpha-tocopherol incorporation into liposome ratios ranged from 0.01 to 12 mol%. Best oxidative stability was obtained for liposomes that contained 5 mol% alpha-tocopherol. Compared to the other formulas, propanal formation was reduced, and time of the oxidation induction phase was longer. Moreover, alpha-tocopherol induced both liposome structural modifications, evidenced by turbidity, and phospholipid chemical hydrolysis, quantified as the amount of lysophospholipids. This physicochemical liposome instability was even more pronounced in acid storage conditions, i.e., alpha-tocopherol incorporation into liposome membranes accelerated the structural rearrangements and increased the rate of phospholipid hydrolysis. In particular, giant vesicles incubated at pH 1.5 underwent complex irreversible shape transformations including invaginations. In parallel, the absorption rate of alpha-tocopherol was measured in lymph-cannulated rats when alpha-tocopherol was administrated, as liposome suspension or added to sardine oil, through a gastrostomy tube. Alpha-tocopherol recovery in lymph was increased by almost threefold, following liposome administration. This may be related to phospholipids that should favor alpha-tocopherol solubilization and to liposome instability in the case of a high amount of alpha-tocopherol in the membranes. A need to correlate results obtained from in vitro liposome behavior with in vivo lipid absorption was demonstrated by this study.

Administration, Oral↗

Cholesterol oxidation in meat from chickens fed alpha-tocopherol- and beta-carotene-supplemented diets with different unsaturation grades.

The production of B-ring and side-chain oxysterols was evaluated in meat from chickens fed diets differing by the kind of oil or fat added. The effect of supplementary levels of natural antioxidants, as alpha-tocopherol and beta-carotene, on the meat cholesterol oxidative stability was also studied. Lard, sunflower and olive oil were used as dietary fat. Raw and cooked meats were analyzed for oxysterols, and cholesterol was also quantified. Oxysterol analyses were carried out by combining the use of solid-phase extraction, thin-layer chromatography, capillary gas chromatography, and capillary gas chromatography-mass spectrometry. Oxysterols were detected within the 0.1-0.5 microg/g range in raw meat. Cooking increased the oxysterol content of the meat, and levels as high as 5 microg/g muscle tissue were observed. B-Ring oxysterols were mainly produced: the alpha- and the beta-epoxycholesterols, the 7alpha- and 7beta-hydroxycholesterols, and the 7-ketocholesterol. The results showed that the meat from the chickens fed the olive oil-based diet containing alpha-tocopherol at 200 mg/kg of diet presented the best cholesterol oxidative stability. A positive effect could not be found for dietary beta-carotene administered at levels of 15 and 50 mg/kg of diet. Furthermore, a significant decrease in the tissue cholesterol content was observed with the olive and the sunflower oil-based diets.

Animal Feed↗

Influence of dietary vitamin E and C supplementation on vitamin E and C content and thiobarbituric acid reactive substances (TBARS) in different tissues of growing pigs.

To investigate the influence and possible interactions of dietary vitamin E and C supplementation on vitamin content of both vitamins and oxidative stability of different pork tissues 40 Large White barrows from 25 kg to 106 kg were allocated to four different cereal based diets: Basal diet (B), dl-alpha-tocopherylacetate + 200 mg/kg (E), crystalline ascorbic acid + 300 mg/kg (C) or both vitamins (EC). At slaughtering samples of liver, spleen, heart, kidney, backfat outer layer, ham and M. tongissimus dorsi were obtained. Growth performance of the pigs and carcass characteristics were not influenced by feeding treatments. Dietary vitamin E supplementation had a significant effect on the vitamin E and alpha-tocopherol concentration in all investigated tissues. Backfat outer layer, liver, spleen, kidney and heart had higher vitamin E concentrations than ham and M. longissimus dorsi. Dietary vitamin C supplementation tended towards enhanced vitamin E levels except for ham samples. Therefore, some synergistic actions without dietary vitamin E supplementation between the two vitamins could be shown. The vitamin C concentration and TBARS were increased or at least equal in all tissues due to vitamin C supplementation. Dietary alpha-tocopherol supplementation resulted in lower TBARS in backfat outer layer (malondialdehyde 0.35 mg/kg in B vs. 0.28 mg/kg in E), but increased in heart and ham. When both vitamins were supplemented (EC) TBARS were lower in M. longissimus dorsi and backfat outer layer, equal in heart and higher in liver and ham compared to a single vitamin C supplementation. Rancimat induction time of backfat outer layer was 0.3 h higher in C compared to B and 0.17 h higher in EC than in E. Correlations between levels of both vitamins were positive for kidney (r = 0.169), M. longissimus dorsi (r = 0.499) and ham (r = 0.361) and negative for heart (r = -0.350). In liver and spleen no interaction could be found. In backfat outer layer vitamin E was positively correlated with rancimat induction time (r = 0.550) and negatively with TBARS (r = -0.202), but provided no evidence that dietary vitamin E supply led to better oxidative stability.

Adipose Tissue↗

Directed evolution of N-carbamyl-D-amino acid amidohydrolase for simultaneous improvement of oxidative and thermal stability.

Directed evolution of N-carbamyl-D-amino acid amidohydrolase from Agrobacterium tumefaciens NRRL B11291 was attempted in order to simultaneously improve oxidative and thermal stability. A mutant library was generated by DNA shuffling, and positive clones with improved oxidative and thermal stability were screened on the basis of the activity staining method on a solid agar plate containing pH indicator (phenol red) and substrate (N-carbamyl-D-p-hydroxyphenylglycine). Two rounds of directed evolution resulted in the best mutant 2S3 with a significantly improved stability. Oxidative stability of the evolved enzyme 2S3 was about 18-fold higher than that of the wild type, and it also showed an 8-fold increased thermostability. The K(m) value of 2S3 was comparable to that of wild-type enzyme, but k(cat) was slightly decreased. DNA sequence analysis revealed that six amino acid residues (Q23L, V40A, H58Y, G75S, M184L, and T262A) were substituted in 2S3. From the mutational analysis, four mutations (Q23L, H58Y, M184L, and T262A) were found to lead to an improvement of both oxidative and thermal stability. Of them, T262A had the most significant effect, and V40A and G75S only increased the oxidative stability.

Agrobacterium tumefaciens↗

Low linolenate and commercial soybean oils diminish serum HDL cholesterol in young free-living adult females.

OBJECTIVE: A mutant soybean line (A16) low in linolenic acid content (2% of oil by weight) was developed to increase oil oxidative stability. It was unknown whether serum lipid and lipoprotein concentrations in humans would be affected should A16 soybean oil (A16 oil) replace commercial soybean oil in diets. This study was conducted to examine the hypothesis that in free-living normolipidemic women, the consumption of A16 oil at approximately 10% of energy intake (en%) would not affect serum lipids and lipoproteins differently than would the consumption of the same amount of a commercial soybean oil with 7% of linolenic acid content. DESIGN: Fifteen free-living female college students consumed the soybean oil daily with regular meals for 9 weeks in different orders, with each test oil being eaten for 3 weeks. During the study, 13 en% was provided by each test oil and a total of 35 en% was from dietary fat. Serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL cholesterol), low-density lipoprotein cholesterol (LDL cholesterol) and triacylglycerides (TAG) were measured. Serum total fatty acid patterns were analyzed as well. RESULTS: Each of the three test oils decreased serum total cholesterol, LDL cholesterol and TAG concentrations from the baseline values. The feeding of A16 and commercial soybean oils decreased serum HDL cholesterol significantly compared with coconut oil (p < 0.05). Dietary inclusion of coconut oil increased serum myristic acid significantly more than did either soybean oil (p < 0.01). Serum arachidonic acid concentrations were significantly greater with A16 consumption than with commercial soybean oil consumption (p < 0.001). CONCLUSION: A16 and commercial soybean oils both diminished serum HDL cholesterol. Although the fatty acid composition differed between the two soybean oils, A16 oil and commercial oil had similar effects on serum concentrations of lipoproteins and lipids. With increased oxidative stability, A16 oil is a good alternative to commercial soybean oil.

Adolescent↗

Stabilization of soybean oil with heated quercetin and 5-caffeoylquinic acid in the presence of ferric ion.

The effect of heated quercetin (400 mg/kg of oil) or 5-caffeoylquinic acid (5-CQA) and the presence of ferric ion (2.2 mg/kg of oil) on the stability of soya oil oxidized in an oxidative stability index (OSI) instrument was investigated. After heating the phenolic at 100 degrees C or 150 degrees C, the OSI values of treated oils were not significantly (p < 0.001) different, whereas, at 200 degrees C, the values decreased significantly with addition of quercetin, but not with 5-CQA. However, the antioxidative activity of quercetin remained significantly greater than that of 5-CQA. The antioxidative potency of quercetin was reduced significantly by addition of ferric palmitate (FP), but that of 5-CQA was not. Reaction between the ortho-dihydroxy groups of the quercetin and ferric ion may reduce the number of hydroxyls available to react with free radicals. Chelating action of 5-CQA might be provided by ortho-dihydroxy grouping of the quinic acid moiety.

Chlorogenic Acid↗