The dual function of nitrite under stomach conditions is modulated by reducing compounds.

Salivary nitrite plays a role in the lipid peroxidation process of muscle tissue in simulated gastric fluid. The objectives of our study were to elucidate the fate of nitrite in the presence of reducing compounds and to evaluate its effect on lipid peroxidation during digestion. Nitrite at pH 3 (possibly NO(2.), not NO.) can oxidize beta-carotene, but the addition of reducing compounds, ascorbic acid or polyphenols, alters its effect. Ascorbic acid alone promoted the formation of NO. from nitrite only up to pH 3, but the addition of iron ions facilitated the formation of NO. up to pH 5.5. NO prevented membranal lipid peroxidation under stomach conditions. Nitrite, only in the presence of reducing compounds, achieved the same goal but at much higher concentrations. Addition of polyphenols to nitrite synergistically improved its antioxidant effect. Therefore, to promote NO. production and to achieve better control of the lipid peroxidation process in the stomach, a nitrite-rich meal should be consumed simultaneously with food rich in polyphenols.

Saliva plays a dual role in oxidation process in stomach medium.

The aim of this study was to evaluate the role of saliva in the oxidation process under the acidic condition of the stomach. Saliva specimens played varied roles in the lipid peroxidation process of heated muscle tissue in simulated gastric fluid: pro-oxidant effects, no effects, and antioxidant effects. To elucidate these differences, selected saliva components were examined. The pseudoperoxidase activity of lactoperoxidase increased lipid peroxidation, while thiocyanate and nitrite-reduced lipid peroxidation. The effect of a saliva specimen on lipid peroxidation was correlated with the concentration of nitrite in the specimen, but not with that of other saliva components. The inhibitory effect of nitrite may be due to its conversion to NO. Elucidation of the antioxidant effect of saliva on co-oxidation of d-alpha-tocopherol in gastric fluid, demonstrated that saliva alone cannot protect d-alpha-tocopherol from co-oxidation, although it partially protected against lipid peroxidation. The presence of red wine polyphenols in stomach medium totally inhibits food lipid peroxidation and d-alpha-tocopherol co-oxidation.

Oxymyoglobin oxidation and membrane lipid peroxidation initiated by iron redox cycle: prevention of oxidation by enzymic and nonenzymic antioxidants.

The red color of muscle is principally due to the presence of oxymyoglobin. Oxidation of heme iron from the ferrous to the ferric state produces a brownish color, which consumers find undesirable. The aim of this study was to use enzymic and nonenzymic antioxidants to simulate in situ muscle antioxidation reactions in order to understand better the mechanism by which the iron redox cycle catalyzes membrane lipid peroxidation and oxymyoglobin oxidation. The inclusion of superoxide dismutase (SOD) in the model system decreased oxymyoglobin oxidation by 10% without affecting lipid peroxidation. Addition of catalase decreased oxymyoglobin oxidation by approximately 40% but not lipid peroxidation. Increasing the ceruloplasmin concentration inhibited lipid peroxidation but increased oxymyoglobin oxidation, which was inhibited by SOD and catalase. Conalbumin (50 microM), a specific iron chelator, inhibited peroxidation and oxymyoglobin oxidation by almost 50%. The addition of the antioxidant catechin (500 microM) decreased lipid peroxidation by 90% but oxymyoglobin oxidation by only 50%. Feeding turkeys with vitamin E at several levels significantly increased the alpha-tocopherol level of membranes, thus preventing oxymyoglobin and lipid oxidation. In conclusion, oxymyoglobin stability in the model system was affected by two pathways: (a) oxygen active species, such as O(2)*(-), H(2)O(2), HO*, and ferryl, generated during autoxidation of myoglobin and oxidation of ferrous ions and ascorbic acid; and (b) lipid radicals, such as ROO*, RO*, and hydroperoxides, generated during lipid peroxidation. Maximum inhibition could be achieved only by introducing inhibitors of both pathways into the system.

Oxymyoglobin oxidation and membranal lipid peroxidation initiated by iron redox cycle.

Oxymyoglobin is the main pigment in muscle tissues, responsible for the bright red color of fresh meat. Oxidation of the heme iron from the ferrous to the ferric metmyoglobin produces the brownish color that consumers find undesirable in fresh meat. The aim of this study was to elucidate the mechanism of oxymyoglobin oxidation in muscle tissues by using a model system containing oxymyoglobin and muscle membranes oxidized by an iron redox cycle. Oxidation of oxymyoglobin was determined from the decrease in absorption of the solution measured by a spectrophotometer at 582 nm. Lipid peroxidation was determined by accumulation of TBARS and conjugated dienes. The higher rates of oxidation of oxymyoglobin (20 microM) and lipid oxidation were achieved by using ferric iron and ascorbic acid at concentrations of 50 and 200 microM, respectively. Increasing the concentration of ascorbic acid to 2000 microM switched its effect to antioxidative. Increasing the concentration of oxymyoglobin from 20 to 80 microM inhibited lipid peroxidation by >90% and partially prevented oxymyoglobin oxidation.

Effects of vitamin E supplementation on lipid peroxidation and color retention of salted calf muscle from a diet rich in polyunsaturated fatty acids.

The color of fresh meat is one of the most important quality criteria of raw muscle foods. This red color is principally due to the presence of oxymyoglobin. The present study was undertaken to examine the effect of a diet rich in polyunsaturated fatty acids (PUFA), the addition of NaCl, and the influence of dietary supplementation with vitamin E on calf muscle oxymyoglobin oxidation (color) and lipid peroxidation. Vitamin E was added to the feed at a concentration of 4000 mg/day for 90 days before slaughter. This diet increased the alpha-tocopherol concentration in muscle membrane from 2.6-2.8 to 6.5-7.0 microg/g of fresh weight. It was found that the diet rich in PUFA and, especially, the addition of NaCl increased muscle lipid peroxidation and oxymyoglobin oxidation as indicated by the contents of thiobarbituric acid-reactive substances and substances that impaired color value readings during storage at 4 degrees C. Both undesirable reactions during storage were controlled very efficiently by the presence of a critically high concentration of alpha-tocopherol in the muscle tissues. The findings concerning the antioxidant activity of alpha-tocopherol in this study form additional evidence of its efficient protection against oxidative reactions during storage of muscle tissues and its potential to maintain a high nutritional value in them.

The stomach as a bioreactor: dietary lipid peroxidation in the gastric fluid and the effects of plant-derived antioxidants.

Atherosclerosis may result partly from processes that occur following food consumption and that involve oxidized lipids in chylomicrons. We investigated reactions that could occur in the acidic pH of the stomach and accelerate the generation of lipid hydroperoxides and co-oxidation of dietary constituents. The ability of dietary polyphenols to invert catalysis from pro-oxidation to antioxidation was examined. The acidic pH of gastric fluid amplified lipid peroxidation catalyzed by metmyoglobin or iron ions. Metmyoglobin catalyzed peroxidation of edible oil, resulting in 8-fold increase of hydroperoxide concentration. The incubation of heated muscle tissue in simulated gastric fluid for 2 h enhanced hydroperoxides accumulation by 6-fold to 1200 microM. In the presence of catechin or red wine polyphenols, metmyoglobin catalyzed the breakdown of hydroperoxides to zero, totally preventing lipid peroxidation and beta-carotene cooxidation. We suggest that human gastric fluid may be an excellent medium for enhancing the oxidation of lipids and other dietary constituents. The results indicate the potentially harmful effects of oxidized fats intake in the presence of endogenous catalysts found in foods, and the major benefit of including in the meal plant dietary antioxidants.

Betalains--a new class of dietary cationized antioxidants.

Antioxidant nutrients from fruits and vegetables are believed to be a class of compounds that exert their effects in humans by preventing oxidative processes which contribute to the onset of several degenerative diseases. This study found a new class of dietary cationized antioxidants in red beets (Beta vulgaris L.). These antioxidants are betalains, and the major one, betanin, is a betanidin 5-O-beta-glucoside. Linoleate peroxidation by cytochrome c was inhibited by betanin, betanidin, catechin, and alpha-tocopherol with IC(50) values of 0.4, 0.8, 1.2, and 5 microM, respectively. In addition, a relatively low concentration of betanin was found to inhibit lipid peroxidation of membranes or linoleate emulsion catalyzed by the "free iron" redox cycle, H(2)O(2)-activated metmyoglobin, or lipoxygenase. The IC(50) inhibition of H(2)O(2)-activated metmyoglobin catalysis of low-density lipoprotein oxidation by betanin was <2.5 microM and better than that of catechin. Betanin and betanidin at very small concentrations were found to inhibit lipid peroxidation and heme decomposition. During this reaction, betanidin was bleached completely, but betanin remained unchanged in its absorption. This difference seems to derive from differing mechanisms of protection by these two compounds. The high affinity of betanin and betanidin for membranes was demonstrated by determining the rate of migration of the compounds through a dialysis tube. Betanin bioavailability in humans was demonstrated with four volunteers who consumed 300 mL of red beet juice, containing 120 mg of the antioxidant. The betacyanins were absorbed from the gut and identified in urine after 2-4 h. The calculated amount of betacyanins found in the urine was 0.5-0.9% of that ingested. Red beet products used regularly in the diet may provide protection against certain oxidative stress-related disorders in humans.

Enhancement of polyphenol recovery from rosemary (Rosmarinus officinalis) and sage (Salvia officinalis) by enzyme-assisted ensiling (ENLAC).

The efficacy of enzyme-assisted ensiling (ENLAC) in the recovery of polyphenols from rosemary and sage was tested. Fresh rosemary and sage were chopped and ensiled in 0.5-L anaerobic jars. Treatments comprised control (no additives), 0.5% glucose and lactic acid bacteria, and 1% cellulase plus 1% hemicellulase plus pectinase. Following storage at room temperature for 45 days (experiment 1) and 26 days (experiment 2), polyphenols were extracted from the silages in ethanol either by direct blending or by cold extraction. The enzyme treatment resulted in silages with the lowest pH values, lowest fiber content, highest water-soluble sugar content, and highest polyphenol recovery; this treatment resulted in increased polyphenol recovery from rosemary and sage, by 100 and 20%, respectively. Comparison between direct blending and cold extraction revealed similar efficiency of polyphenol recovery.

PH-dependent forms of red wine anthocyanins as antioxidants.

Anthocyanins are one of the main classes of flavonoids in red wines, and they appear to contribute significantly to the powerful antioxidant properties of the flavonoids. In grapes and wines the anthocyanins are in the flavylium form. However, during digestion they may reach higher pH values, forming the carbinol pseudo-base, quinoidal-base, or the chalcone, and these compounds appear to be absorbed from the gut into the blood system. The antioxidant activity of these compounds, in several metal-catalyzed lipid oxidation model systems, was evaluated in comparison with other antioxidants. The pseudo-base and quinoidal-base malvidin 3-glucoside significantly inhibited the peroxidation of linoleate by myoglobin. Both compounds were found to work better than catechin, a well-known antioxidant. In a membrane lipid peroxidation system, the effectiveness of the antioxidant was dependent on the catalyst: In the presence of H(2)O(2)-activated myoglobin, the inhibition efficiency of the antioxidant was malvidin 3-glucoside > catechin > malvidin > resveratrol. However, in the presence of an iron redox cycle catalyzer, the order of effectiveness was resveratrol > malvidin 3-glucoside = malvidin > catechin. The pH-transformed forms of the anthocyanins remained effective antioxidants in these systems, and their I(50) values were between 0.5 and 6.2 microM.

Effect of high levels of dietary iron, iron injection, and dietary vitamin E on the oxidative stability of turkey meat during storage.

A study was carried out to evaluate the combined effect of excess Fe, supplied either in the diets (Experiments 1, 2, and 3) or by injection (Experiment 4), and various levels of dietary vitamin E on the oxidative stability of the thigh muscle of turkeys stored at -18 C for various periods. Iron was added to a commercial diet that already contained 20 mg/kg supplemental Fe, at concentrations of 0, 100, 250, and 500 mg/kg as ferrous sulfate or injected as Fe-dextran to the left drumstick muscle (total amount of 1.2 g per turkey). Vitamin E was added to the experimental diets not already supplemented with this vitamin, at levels of 0, 28, and 150 mg/kg. Thiobarbituric acid reactive substances (TBARS) values of the meat gradually increased as its storage duration increased from about 15 to 120 d. Increasing dietary Fe supplementation from 0 to 500 mg/kg tended to decrease TBARS values in one experiment only; otherwise, this variable was not affected by dietary Fe level. Injection of Fe significantly (P < 0.05) increased TBARS values, only in meat from the injected side. The TBARS values of the meat up to about 30 d of storage were significantly lower due to the supplementation of the diet with vitamin E at a level of 28 mg/kg in one out of three experiments and at a level of 150 mg/kg in two out of two experiments. The protective effect of the higher level of vitamin E remained evident after about 108 d of storage. No interaction was observed between Fe and vitamin E treatments in their effect on TBARS values. Blood hemoglobin concentrations were significantly increased by the supplementation of the diet with the high levels of Fe, in one experiment only. This variable was consistently and significantly increased from about 10 to 23 wk of age. The results show that high levels of dietary Fe do not adversely affect the oxidative stability of thigh meat of turkey; however, stability might be reduced by injected Fe. Dietary vitamin E, at a level of 150 mg/kg, consistently increased this stability.

Factors that interact with the antibacterial action of thyme essential oil and its active constituents.

The viable counts of Salmonella typhimurium on nutrient agar (NA) decreased upon the addition of either the essential oil of thyme or its constituent thymol, especially under anaerobic conditions. Antagonistic effects of thymol against Staphylococcus aureus were also greater under anaerobic conditions. In contrast to the phenolic constituents of the oil, thymol and carvacrol, the chemically related terpenes p-cymene and gamma-terpinene had no antagonistic effects against Salm. typhimurium. The addition of Desferal to NA counteracted the antibacterial effects of both thyme oil and thymol. No support was obtained, however, for a possible role of iron in the oxygen-related antibacterial action of the thyme oil and thymol or for the observed effect of Desferal. In the presence of thymol, the viable counts of Salm. typhimurium obtained on a minimal medium (MM) were lower than those obtained on NA. Addition of bovine serum albumin (BSA) neutralized the antibacterial action of thymol. It is suggested that the effects of BSA or Desferal are due to their ability to bind phenolic compounds through their amino and hydroxylamine groups, respectively, thus preventing complexation reactions between the oil phenolic constituents and bacterial membrane proteins. This hypothesis is supported by the marked decrease in the viable counts of Salm. typhimurium caused by either thyme oil or thymol when the pH of the medium was changed from 6.5 to 5.5 or the concentration of Tween 80 in the medium was reduced.

Oxidative processes in meat and meat products: Quality implications.

Lipid peroxidation is, in most instances, a free radical chain reaction that can be described in terms of initiation, propagation, branching and termination processes. With regard to lipid peroxidation, one of the most important questions concerns the source of the primary catalysts that initiate peroxidation in situ in muscle foods. When cells are injured, such as in muscle foods after slaughtering, lipid peroxidation is favored, and traces of O(2) and H(2)O(2), indicating lipid peroxides, are formed. The stability of a muscle food product will depend on the 'tone' of these peroxides and especially from the involvement of metal ions in the process. The cytosol contains not only prooxidants but also antioxidants and the tone of both affects the overall oxidation. Lipid peroxidation is one of the primary mechanisms of quality deterioration in foods and especially in meat products. The changes in quality can be manifested by deterioration in flavor, color, texture, nutritive value and the production of toxic compounds.

Inhibition of oxidation of human low-density lipoprotein by phenolic substances in red wine.

The "French paradox" (apparent compatibility of a high fat diet with a low incidence of coronary atherosclerosis) has been attributed to the regular drinking of red wine. However, the alcohol content of wine may not be the sole explanation for this protection. Red wine also contains phenolic compounds, and the antioxidant properties of these may have an important role. In in-vitro studies with phenolic substances in red wine and normal human low-density lipoprotein (LDL) we found that red wine inhibits the copper-catalysed oxidation of LDL. Wine diluted 1000-fold containing 10 mumol/L total phenolics inhibited LDL oxidation significantly more than alpha-tocopherol. Our findings show that the non-alcoholic components of red wine have potent antioxidant properties toward oxidation of human LDL.

Nitric oxide, an inhibitor of lipid oxidation by lipoxygenase, cyclooxygenase and hemoglobin.

The present study demonstrated that nitric oxide, which is an important mammalian metabolite, can inhibit oxidation by lipoxygenase, cyclooxygenase and hemoglobin. The inhibition is manifested as a lag-phase that is reversible. The inhibitory effect of nitric oxide on lipoxygenase and cyclooxygenase seems to derive from i) the capability of .NO to reduce the ferric enzyme to the ferrous form, which is inactive; ii) competition for the iron site available for exogenous ligands; and iii) the radical scavenging ability of the nitroxide radical. Nitric oxide may act as a modulator of the arachidonic acid cascade and in the generation of oxygen-active species.

Nitric oxide as an antioxidant.

Benzoate monohydroxy compounds, and in particular salicylate, were produced during interaction of ferrous complexes with hydrogen peroxide (Fenton reaction) in a N2 environment. These reactions were inhibited when Fe complexes were flushed, prior to the addition in the model system, by nitric oxide. Methionine oxidation to ethylene by Fenton reagents was also inhibited by nitric oxide. Myoglobin in several forms such as metmyoglobin, oxymyoglobin, and nitric oxide-myoglobin were interacted with an equimolar concentration of hydrogen peroxide. Spectra changes in the visible region and the changes in membrane (microsomes) lipid peroxidation by the accumulation of thiobarbituric acid-reactive substances (TBA-RS) were determined. The results showed that metmyoglobin and oxymyoglobin were activated by H2O2 to ferryl myoglobin, which initiates membrane lipid peroxidation; but not nitric oxide-myoglobin, which, during interaction with H2O2, did not form ferryl but metmyoglobin which only poorly affected lipid peroxidation. It is assumed that nitric oxide, liganded to ferrous complexes, acts to prevent the prooxidative reaction of these complexes with H2O2.

Haemoglobin and myoglobin as inhibitors of hydroxyl radical generation in a model system of "iron redox" cycle.

Methionine was oxidized to ethylene by an "Iron Redox" system containing H2O2, Fe-EDTA and ascorbate, generating hydroxyl radicals or another species of similar reactivity. Oxy or met forms of haemoglobin and myoglobin were found to inhibit methionine oxidation. Methionine oxidation was elevated in the "Iron Redox" system by increasing ascorbic acid concentration. However, in the presence of metmyoglobin or methaemoglobin, the increases in ascorbic acid did not lower the haemproteins' inhibitory effects but rather increased them. The pro-oxidative or anti-oxidative activities of haemproteins in biological oxidative reactions seem to be dependent on compartmentalization and on the presence and concentrations of reducing compounds and H2O2.

Iron release from metmyoglobin, methaemoglobin and cytochrome c by a system generating hydrogen peroxide.

The reaction of H2O2 with resting metmyoglobin (MetMb), methaemoglobin (MetHb) and cytochrome-c (Cyt-c) was studied in the Soret and visible regions. The differences between the original and the final peak heights of the native haemproteins at 408 nm was found to be directly proportional to the loss of iron from the molecule. The release of iron from haemproteins was studied in a system generating H2O2 continuously at a low rate by an enzymic system, or by addition of large amounts of H2O2. Cytochrome-c, methaemoglobin and metmyoglobin during interaction with H2O2 at a concentration of 200 microM release 40%, 20% and 3%, respectively, of molecular iron after 10 min. The inhibition of haem degradation and iron release by enzymatically-generated H2O2 was determined using several hydroxyl radical scavengers, reducing agents and antioxienzymes, such as superoxide dismutase, catalase and caeruloplasmin.