Meat crust as a novel food ingredient to regulate lipid peroxidation and oxidative stress.

Pan Fry (PF) is a common heating treatment however, there is limited data on meat oxidation after PF using direct contact with an uncoated iron pan. After PF, a crust is formed, and in this study, we aim to evaluate the potential anti-oxidation and anti-lipid peroxidation capacity of such crust. Ground beef and turkey meat were heat treated using PF or microwave. Lipid peroxidation was evaluated using malondialdehyde accumulation. PF meat generated lower lipid peroxidation levels versus microwave-heated meat. Iron PF has decreased lipid peroxidation versus Teflon pan heating. The crust significantly lowered lipid peroxidation and possessed millard reaction products (MRPs), strong reducing abilities, iodine removal capacity, and some iron chelation capacity. We demonstrated that the crust substantially decreases lipid peroxidation levels in various systems and can be used as a novel seminatural antioxidant ingredient, which may lead to extended shelf life and protects various food products.

Food Polyphenols as Preventive Medicine.

Reactive oxygen species (ROS) are the initiators in foods and in the stomach of oxidized dietary lipids, proteins, and lipid-oxidation end-products (ALEs), inducing in humans the development of several chronic diseases and cancer. Epidemiological, human clinical and animal studies supported the role of dietary polyphenols and derivatives in prevention of development of such chronic diseases. There is much evidence that polyphenols/derivatives at the right timing and concentration, which is critical, acts mostly in the aerobic stomach and generally in the gastrointestinal tract as reducing agents, scavengers of free radicals, trappers of reactive carbonyls, modulators of enzyme activity, generators of beneficial gut microbiota and effectors of cellular signaling. In the blood system, at low concentration, they act as generators of electrophiles and low concentration of H2O2, acting mostly as cellular signaling, activating the PI3K/Akt-mediated Nrf2/eNOS pathways and inhibiting the inflammatory transcription factor NF-κB, inducing the cells, organs and organism for eustress, adaptation and surviving.

DMF-Activated Nrf2 Ameliorates Palmitic Acid Toxicity While Potentiates Ferroptosis Mediated Cell Death: Protective Role of the NO-Donor S-Nitroso-N-Acetylcysteine.

Nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease that can develop into an aggressive form called nonalcoholic steatohepatitis (NASH), which ultimately progresses to cirrhosis, hepatocellular carcinoma (HCC), and end-stage liver failure. Currently, the deterioration of NAFLD is attributed to specific lipid toxicity which could be due to lipotoxicity and/or ferroptosis. In the current study, we evaluated the involvement of the nuclear factor erythroid 2 (NFE2)-related factor 2 (Nrf-2), which is a main activator of phase II metabolism in the two types of lipid-induced toxicity in hepatocytes, lipotoxicity by saturated fatty acids, and in ferroptosis, and the effect of NO donor treatment. AML12 cells were exposed to 600 µM palmitic acid to induce lipotoxicity or treated with 20 µM erastin or 5 µM RSL3 for ferroptosis. In SFA-lipotoxicity, pretreatment with the Nrf2 activator dimethyl fumarate (DMF) managed to ameliorate the cells and the oxidative stress level while aggravating ferroptosis due to emptying the thiol pool. On the other hand, the nitric oxide (NO)-donor, S-nitroso-N-acetylcysteine (NAC-SNO) proved to be effective in the prevention of hepatocytes ferroptosis.

Reactivity of Vitamin E as an Antioxidant in Red Meat and the Stomach Medium.

The stomach is a bioreactor and an important intersection of biochemical reactions that affect human health. Lipid peroxidation of meat in the stomach medium generates malondialdehyde (MDA), which is absorbed from the gut into human plasma and modifies low-density lipoprotein (LDL) to MDA-LDL. We found in the stomach medium (pH 3.0) a high antioxidant activity of vitamin E against meat lipid peroxidation, almost 35-fold higher than at pH 6.3. In the stomach medium, the antioxidant activity of vitamin E on meat lipid peroxidation was 20-fold higher than that of catechin. Vitamin E, at pH 3.0, acts synergistically with metmyoglobin (MbFe+3), as a peroxidase/antioxidant couple. The synergistic effect of MbFe+3/vitamin E was almost 150-fold higher than the antioxidant effect achieved by MbFe+3/catechin. The meat antioxidant activity was maintained continuously by addition of a low concentration of vitamin E, catechin, and vitamin C, preventing the propagation of lipid oxidation, reactive aldehyde generation, and the loss of vitamin E.

S-Nitroso-N-acetylcysteine (NAC-SNO) vs. nitrite as an anti-clostridial additive for meat products.

Nitrite is added to meat products as a preservative and it acts as a bacteriostatic compound against Clostridium botulinum growth. Nitric-oxide (˙NO), myoglobin and S-nitroso-compounds seem to be the main molecules generated from nitrite in meat products, which by decomposition to ˙NO, form the main anti-clostridial factor. The growth of C. sporogenes from activated spores in the presence of 0.5-2.5 mM NAC-SNO was compared to nitrite, both at 37 °C for 5 days and at room temperature for 28 days. The present study demonstrates that NAC-SNO under the same conditions and concentrations, in meat products, acts as an anti-clostridial compound similar to nitrite. In contrast to nitrite which must be activated in meat by heating, NAC-SNO generates the anti-clostridial factor directly, without heating, as was evaluated in an unheated bacteriological medium. The toxic effect of NAC-SNO and nitrite in methaemoglobinaemia and generation of N-nitrosamines in vivo, in mice, were also determined. Mice were gavage fed milk containing 45 mg per kg per bw of nitrite or an equimolar equivalent of NAC-SNO in the presence of 50 mg per kg per bw of N-methylaniline. Nitrite generated methaemoglobinaemia and carcinogenic N-nitrosoamines (N-nitrosomethylaniline); however, NAC-SNO under the same conditions and concentrations generates much less methaemoglobin and no detectable N-nitrosoamines in the blood, in vivo.

Polyphenols by Generating H2O2, Affect Cell Redox Signaling, Inhibit PTPs and Activate Nrf2 Axis for Adaptation and Cell Surviving: In Vitro, In Vivo and Human Health.

Human health benefits from different polyphenols molecules consumption in the diet, derived mainly by their common activities in the gastrointestinal tract and at the level of blood micro-capillary. In the stomach, intestine and colon, polyphenols act as reducing agents preventing lipid peroxidation, generation and absorption of AGEs/ALEs (advanced glycation end products/advanced lipid oxidation end products) and postprandial oxidative stress. The low absorption of polyphenols in blood does not support their activity as antioxidants and their mechanism of activity is not fully understood. The results are from in vitro, animal and human studies, detected by relevant oxidative stress markers. The review carries evidences that polyphenols, by generating H2O2 at nM concentration, exogenous to cells and organs, act as activators of signaling factors increasing cell Eustress. When polyphenols attain high concentration in the blood system, they generate H2O2 at µM concentration, acting as cytotoxic agents and Distress. Pre-treatment of cells or organisms with polyphenols, by generating H2O2 at low levels, inhibits cellular PTPs (protein tyrosine phosphatases), inducing cell signaling through transcription of the Nrf2 (nuclear factor erythroid 2-related factor 2) axis of adaptation and protection to oxidation stress. Polyphenols ingestion at the right amount and time during the meal acts synergistically at the level of the gastrointestinal tract (GIT) and blood system, for keeping the redox homeostasis in our organism and better balancing human health.

S-Nitroso-N-acetylcysteine (NAC-SNO) as an Antioxidant in Cured Meat and Stomach Medium.

The stability of lipids in meat products depends on the initial concentration of hydroperoxides, the catalytic involvement of metal ions and myoglobin, endogenous antioxidants, and biological and technological factors. Ground meat was treated with additives, sealed in vacuum bags, heated to 75 °C, and stored opened to air at 4 °C. S-Nitroso-N-acetylcysteine (NAC-SNO) at concentration like nitrite used by the industry prevents lipid peroxidation in the product, even after storage for 1 month at 4 °C. The same simulated treatments at different concentrations of both compounds show that NAC-SNO acts as an antioxidant ∼4-fold better than nitrite at pH 6.2 or 3.0. Ascorbic acid significantly improves nitrite antioxidant effect. NAC-SNO was found to prevent, much better than nitrite, accumulation of reactive aldehydes and hydroxynonenal protein modification. In condition like those used by the industry for meat products processing, NAC-SNO acts better than nitrite to provide antioxidant protection without the side effect of N-nitrosation, oxidation, and the loss of nutrient generated by nitrite.

S-Nitroso-N-acetylcysteine Generates Less Carcinogenic N-Nitrosamines in Meat Products than Nitrite.

Nitrite reacts with secondary amines to form N-nitrosamines (N-NA), which lead to gastrointestinal cancers. The aim of this study was to compare nitrite with S-nitrosocysteine (Cys-SNO) and S-nitroso-N-acetylcysteine (NAC-SNO) with respect to N-NA formation, which was evaluated by determining the conversion of N-methylaniline to N-nitrosomethylaniline. Under neutral and acidic pH conditions, N-NA formation rate was nitrite > Cys-SNO > NAC-SNO. In the presence of copper or nucleophiles, NAC-SNO generated much less N-NA than Cys-SNO. Nitrite and Cys-SNO produced higher amounts of N-NA in the presence of oxygen, whereas NAC-SNO was almost oxygen insensitive. In meat in the stomach medium, NAC-SNO produced much lower amounts of N-NA than other additives. In heated meat, Cys-SNO and NAC-SNO generated the nitrosyl-hemochrome pink pigment, better than nitrite. In conclusion, NAC-SNO was much less reactive for N-NA formation than nitrite and Cys-SNO in conditions relevant to meat production and stomach digestion.

Redox homeostasis in stomach medium by foods: The Postprandial Oxidative Stress Index (POSI) for balancing nutrition and human health.

Red-meat lipid peroxidation in the stomach results in postprandial oxidative stress (POS) which is characterized by the generation of a variety of reactive cytotoxic aldehydes including malondialdehyde (MDA). MDA is absorbed in the blood system reacts with cell proteins to form adducts resulting in advanced lipid peroxidation end products (ALEs), producing dysfunctional proteins and cellular responses. The pathological consequences of ALEs tissue damage include inflammation and increased risk for many chronic diseases that are associated with a Western-type diet. In earlier studies we used the simulated gastric fluid (SGF) condition to show that the in vitro generation of MDA from red meat closely resembles that in human blood after consumption the same amount of meat. In vivo and in vitro MDA generations were similarly suppressed by polyphenol-rich beverages (red wine and coffee) consumed with the meal. The present study uses the in vitro SGF to assess the capacity of more than 50 foods of plant origin to suppress red meat peroxidation and formation of MDA. The results were calculated as reducing POS index (rPOSI) which represents the capacity in percent of 100g of the food used to inhibit lipid peroxidation of 200g red-meat a POSI enhancer (ePOSI). The index permitted to extrapolate the need of rPOSI from a food alone or in ensemble such Greek salad, to neutralize an ePOSI in stomach medium, (ePOS-rPOSI=0). The correlation between the rPOSI and polyphenols in the tested foods was R2=0.75. The Index was validated by comparison of the predicted rPOSI for a portion of Greek salad or red-wine to real inhibition of POS enhancers. The POS Index permit to better balancing nutrition for human health.

Lipid Peroxidation in a Stomach Medium Is Affected by Dietary Oils (Olive/Fish) and Antioxidants: The Mediterranean versus Western Diet.

Red meat is an integral part of the Western diet, and high consumption is associated with an increased risk of chronic diseases. Using a system that simulated the human stomach, red meat was interacted with different oils (olive/fish) and lipid peroxidation was determined by measuring accumulation of malondialdehyde (MDA) and lipid peroxides (LOOH). Olive oil decreased meat lipid peroxidation from 121.7 ± 3.1 to 48.2 ± 1.3 µM and from 327.1 ± 9.5 to 77.3 ± 6.0 µM as assessed by MDA and ROOH, respectively. The inhibitory effect of olive oil was attributed to oleic acid rather than its polyphenol content. In contrast, fish oils from tuna or an ω-3 supplement dramatically increased meat lipid peroxidation from 96.2 ± 3.6 to 514.2 ± 6.7 µM MDA. Vitamin E inhibited meat lipid peroxidation in the presence of olive oil but paradoxically increased peroxidation in the presence of fish oil. The inhibitory properties of oleic acid may play a key role in the health benefits of the Mediterranean diet.

Coffee polyphenols protect human plasma from postprandial carbonyl modifications.

The antioxidant capability of coffee polyphenols to inhibit red-meat lipid peroxidation in stomach medium and absorption into blood of malondialdehyde (MDA) in humans was studied. Roasted-ground coffee polyphenols that were found to inhibit lipid peroxidation in stomach medium are 2- to 5-fold more efficient antioxidant than those found in instant coffee. Human plasma from ten volunteers analyzed after a meal of red-meat cutlets (250 g) revealed a rapid accumulation of MDA. The accumulation of MDA in human plasma modified low-density lipoprotein is known to trigger atherogenesis. Consumption of 200 mL roasted coffee by ten volunteers during a meal of red-meat cutlets, resulted after 2 and 4 h in the inhibition by 80 and 50%, respectively, of postprandial plasma MDA absorption. The results obtained in vitro simulated stomach model on MDA accumulation were predictive for the amount of MDA absorbed into circulating human plasma, in vivo. Timing the consumption of coffee during the meals may make it a very active functional food.

Saliva increases the availability of lipophilic polyphenols as antioxidants and enhances their retention in the oral cavity.

OBJECTIVE: Lipophilic polyphenols in fruit beverages can avidly bind to surfaces of microorganisms and to blood cells and to impart upon them enhanced oxidant scavenging abilities (OSA). However, since many of the polyphenols are actually not fully soluble in water, they are therefore not available to act as effective antioxidant agents. We hypothesized that whole saliva, proteins such as albumin and mucin, human red blood cells and platelets, may all increase the "solubility" and availability of lipophilic antioxidant polyphenols thus increasing the OSA of whole saliva. DESIGN: The OSA of whole un-stimulated human saliva, obtained from healthy donors and of combinations among saliva, mucin, blood cells, fruit beverages and reagent polyphenols were quantified by chemiluminescence, DPPH radical and tetrazolium reduction assays. Kinetics of the clearance of polyphenols from saliva after holding in the mouth for 30s of an extract from beverages cinnamon was assayed by the Folin Ciocalteu's and the luminescence assays. RESULTS: OSA of fruit beverages and of reagent polyphenols were markedly increased by whole saliva, mucin and by red blood cells. Polyphenols associated with a cinnamon extract were retained in the oral cavity for several hours as measured by luminescence and Folin reagent techniques. CONCLUSIONS: A new approach to explain the additional role of saliva and salivary proteins and of blood cells as enhancers of OSA of lipophilic polyphenols is presented. This might have a significant importance to assess complex interactions among polyphenols from nutrients, salivary antioxidants, salivary proteins and blood cells extravasated from injure capillaries during infection and inflammation.

Protection by polyphenols of postprandial human plasma and low-density lipoprotein modification: the stomach as a bioreactor.

Recent studies dramatically showed that the removal of circulating modified low-density lipoprotein (LDL) results in complete prevention of atherosclerosis. The gastrointestinal tract is constantly exposed to food, some of it containing oxidized compounds. Lipid oxidation in the stomach was demonstrated by ingesting heated red meat in rats. Red wine polyphenols added to the rats' meat diet prevented lipid peroxidation in the stomach and absorption of malondialdehyde (MDA) in rat plasma. In humans, postprandial plasma MDA levels rose by 3-fold after a meal of red meat cutlets. MDA derived from meat consumption caused postprandial plasma LDL modification in human. The levels of plasma MDA showed a 75% reduction by consumption of red wine polyphenols during the meat meal. Locating the main biological site of action of polyphenols in the stomach led to a revision in the understanding of how antioxidants work in vivo and may help to elucidate the mechanism involved in the protective effects of polyphenols in human health.

Polyphenols activate Nrf2 in astrocytes via H2O2, semiquinones, and quinones.

Polyphenols, which occur both in edible plants and in foodstuff, have been reported to exert a wide range of health effects; however, the mechanism of action of these molecules is not fully understood. One important cellular pathway affected by polyphenols is the activation of the transcription factor Nrf2 via the electrophile response element, which mediates generation of phase 2 detoxifying enzymes. Our study found that Nrf2 nuclear translocation and the activity of NAD(P)H quinone oxidoreductase (NQO1) were increased significantly after treatment of astrocytes with tert-butylhydroquinone (tBHQ), resveratrol, or curcumin, at 20-50µM. Incubation of tBHQ, resveratrol, and curcumin in the growth medium in the absence of astrocytes caused the accumulation of H(2)O(2). Treatment of cells with either glutathione or metmyoglobin was found to decrease Nrf2 translocation and NQO1 activity induced by polyphenols by up to 40 and 60%, respectively. Addition of both glutathione and metmyoglobin to growth medium decreased Nrf2 translocation and NQO1 activity by up to 100 and 80%, respectively. In conclusion, because metmyoglobin, in the presence of polyphenols and glutathione, is known to interact with H(2)O(2), semiquinones, and quinones, the up-regulation of the antioxidant defense of the cells through activation of the Nrf2 transcription factor, paradoxically, occurs via the generation of H(2)O(2) and polyphenol-oxidized species generated from the exogenous microenvironment of the cells.

The stomach as a "bioreactor": when red meat meets red wine.

To determine the stomach bioreactor capability for food oxidation or antioxidation, rats were fed red turkey meat cutlets (meal A) or red turkey meat cutlets and red wine concentrate (meal B). The hydroperoxides (LOOH) and malondialdehyde (MDA) levels of the stomach contents were evaluated during and after digestion; the postprandial plasma MDA level was also evaluated. In independently fed rats, the stomach LOOH concentration fell substantially 90 min following the meal, and the addition of red wine polyphenols enhanced LOOH reduction 3-fold. A similar trend was obtained for MDA. After pyloric ligation, the stomach contents of rats fed red meat homogenate showed >2-fold increases in LOOH and MDA accumulation. The postprandial plasma MDA level increased significantly by 50% following meal A and was maintained or even fell by 34% below basal level following meal B. The findings show that consumption of partially oxidized food could increase lipid peroxidation in the stomach and the absorption of cytotoxic lipid peroxidation products into the body. The addition of antioxidants such as red wine polyphenols to the meal may alter these outcomes. These findings explain the potentially harmful effects of oxidized fats in foods and the important benefit of consuming dietary polyphenols during the meal.

A novel function of red wine polyphenols in humans: prevention of absorption of cytotoxic lipid peroxidation products.

Current evidence supports a contribution of polyphenols to the prevention of cardiovascular disease, but their mechanisms of action are not understood. We investigated the impact of red wine polyphenols on postprandial cytotoxic lipid peroxidation products (MDA) levels in humans. In a randomized, crossover study, the effect of red wine polyphenols on postprandial levels of plasma and urine MDA was investigated. Three meals of 250 g turkey cutlets supplemented by water (A); soaked in red wine after heating plus 200 ml of red wine (B); or soaked in red wine prior to heating plus 200 ml of red wine (C) were administered to 10 healthy volunteers. Subject baseline plasma levels of MDA were 50 +/- 20 nM. After a meal of turkey meat cutlets, plasma MDA levels increased by 160 nM (P<0.0001); after (B) there was a 75% reduction in the absorption of MDA (P<0.0001). However, after (C), the elevation of plasma MDA was completely prevented (P<0.0001). Similar results were obtained for MDA accumulation in urine. Our study suggests that red wine polyphenols exert a beneficial effect by the novel new function, absorption inhibition of the lipotoxin MDA. These findings explain the potentially harmful effects of oxidized fats found in foods and the important benefit of dietary polyphenols in the meal.

Dietary advanced lipid oxidation endproducts are risk factors to human health.

Lipid oxidation in foods is one of the major degradative processes responsible for losses in food quality. The oxidation of unsaturated fatty acids results in significant generation of dietary advanced lipid oxidation endproducts (ALEs) which are in part cytotoxic and genotoxic compounds. The gastrointestinal tract is constantly exposed to dietary oxidized food compounds, after digestion a part of them are absorbed into the lymph or directly into the blood stream. After ingestion of oxidized fats animals and human have been shown to excrete in urine increase amounts of malondialdehyde but also lipophilic carbonyl compounds. Oxidized cholesterol in the diet was found to be a source of oxidized lipoproteins in human serum. Some of the dietary ALEs, which are absorbed from the gut to the circulatory system, seems to act as injurious chemicals that activate an inflammatory response which affects not only circulatory system but also organs such as liver, kidney, lung, and the gut itself. We believe that repeated consumption of oxidized fat in the diet poses a chronic threat to human health. High concentration of dietary antioxidants could prevent lipid oxidation and ALEs generation not only in foods but also in stomach condition and thereby potentially decrease absorption of ALEs from the gut. This could explains the health benefit of diets containing large amounts of dietary antioxidants such those present in fruits and vegetables, or products such as red-wine or tea consuming during the meal.

Lipid peroxidation by "free" iron ions and myoglobin as affected by dietary antioxidants in simulated gastric fluids.

Grilled red turkey muscle (Doner Kabab) is a real "fast food" containing approximately 200 microM hydroperoxides, homogenized in simulated gastric fluid and oxidized more rapidly at pH 3.0 than at pH 5.0, after 180 min, producing 1200 and 600 microM hydroperoxides, respectively. The effects of "free" iron ions and metmyoglobin, two potential catalyzers of lipid peroxidation in muscle foods, were evaluated for linoleic acid peroxidation at pH 3.0 of simulated gastric fluid. The prooxidant effects of free iron ions on linoleic acid peroxidation in simulated gastric fluid was evaluated in the presence of ascorbic acid. At low concentrations of ascorbic acid, the effects were prooxidative, which was reversed at high concentrations. In the presence of metmyoglobin, ascorbic acid with or without free iron enhanced the antioxidative effect. Lipid peroxidation by an iron-ascorbic acid system was inhibited totally by 250-500 microM catechin at pH 3.0. The catechin antioxidant effect was determined also in the iron-ascorbic acid system containing metmyoglobin. In this system, catechin totally inhibited lipid peroxidation at a concentration 20-fold lower than without metmyoglobin. The ability of catechin to inhibit lipid peroxidation was also determined at a low pH with beta-carotene as a sensitive target molecule for oxidation. The results show that a significant protection was achieved only with almost 100-fold higher antioxidant concentration. Polyphenols from different groups were determined for the antioxidant activity at pH 3.0. The results show a high antioxidant activity of polyphenols with orthodihydroxylated groups at the B ring, unsaturation, and the presence of a 4-oxo group in the heterocyclic ring, as demonstrated by quercetin.

Lipid hydroperoxidase activity of myoglobin and phenolic antioxidants in simulated gastric fluid.

Our recent study demonstrated the potential of gastric fluid at pH 3.0 to accelerate lipid peroxidation and cooxidation of dietary constituents in the stomach medium. Metmyoglobin is known to catalyze the breakdown of lipid hydroperoxides to free radicals, a reaction that could enhance the propagation step and general lipid peroxidation. During this reaction, a part of the free radicals is autoreduced by metmyoglobin. At pH 3.0, metmyoglobin at low concentration was almost 7 x 10(4) times as effective as at pH 7.0 in enhancing the rate of lipid peroxidation. Our study demonstrated that metmyoglobin, at a low concentration (approximately 1:30), as compared with that of the hydroperoxides in the lipid system, worked prooxidatively increasing the amounts of linoleate hydroperoxides. However, at a high concentration (approximately 1:3), metmyoglobin acted antioxidatively and decomposed hydroperoxides, whose concentrations then remained at zero for a long time. Catechin, a known polyphenol, supports the inversion of metmyoglobin catalysis, from prooxidation to antioxidation. The antioxidative activity of the couple metmyoglobin-catechin was better at pH 3.0 than at pH 7.0, indicating that this reaction is more dependent on metmyoglobin than on catechin. During this reaction, catechin or quercetin not only donates reducing equivalents to prevent lipid peroxidation but also prevents the destruction and polymerization of metmyoglobin. The results of this research highlighted the important and possible reactions of heme proteins and polyphenols as couple antioxidants, working as hydroperoxidases or as pseudo-peroxidases. We hypothesize that the occurrence of these reactions in the stomach could have an important impact on our health and might help to better explain the health benefits of including foods rich in polyphenol antioxidants in the meal, especially when consuming red meat.