Mitochondrial oxygen consumption in permeabilized fibers and its link to colour changes in bovine M. semimembranosus muscle.

Animal and muscle characteristics were recorded for 41 cattle. The oxygen consumption rate (OCR) of M. semimembranosus was measured between 3.0-6.4h post mortem (PM3-6) and after 3 weeks in a vacuum pack at 4°C. Colour change measurements were performed following the 3 weeks using reflectance spectra (400-1,100 nm) and the colour coordinates L, a and b, with the samples being packaged in oxygen permeable film and stored at 4°C for 167 h. Significant individual animal differences in OCR at PM3-6 were found for mitochondrial complexes I and II. OCR of complex I declined with increased temperature and time PM, while residual oxygen-consuming side-reactions (ROX) did not. OCR of stored muscles was dominated by complex II respiration. A three-way regression between samples, colour variables collected upon air exposure and OCR of 3 weeks old fibres revealed a positive relationship between OCR and complex II activity and also between OCR and OCR(ROX). The presence of complex I and ß-oxidation activities increased metmyoglobin formation.

Use of dietary vitamin E and selenium (Se) to increase the shelf life of modified atmosphere packaged light lamb meat.

The aim of this work was to determine the increase in the shelf life of modified atmosphere packaged fresh lamb meat due to the effect of dietary vitamin E and selenium supplementation on colour and lipid oxidation. 128 lambs were fed on a concentrate with standard levels of vitamin E (C), a concentrate enriched with vitamin E (V), a concentrate with sodium selenite (S) and a concentrate enriched with both vitamin E and sodium selenite (VS). The lambs were slaughtered at 27.3±1.45 kg LW, and chops stored on MAP for 7, 9, 11 and 13 days. CIELab colour and TBARs were studied on these days. Use of dietary vitamin E extended the shelf life a further 4 days from the commercial sell-by date in terms of lightness, hue angle, metmyoglobin formation and lipid oxidation. Selenium could be used to increase the lightness of meat without vitamin E supplementation in lambs' diets.

Effects of malate, lactate, and pyruvate on myoglobin redox stability in homogenates of three bovine muscles.

We investigated the effects of glycolytic and tricarboxylic acid cycle metabolic intermediates on myoglobin redox forms and meat colour stability. Eighteen combinations of malate (M), lactate (L), and pyruvate (P) were added to beef Longissimus lumborum, Psoas major, and Semitendinosus muscle homogenates to study their effect on metmyoglobin formation during incubation at 25 degrees C. Changes in surface colour at 0, 2, 4, 8, and 12 h were evaluated by using reflecto-spectrophotometry [both L*, a*, and b* and wavelengths specific for metmyoglobin (MMb)]. Addition of M, L, and P alone or in combinations stabilized (P < 0.05) L*, a*, and b* values and myoglobin redox forms in muscle homogenates; however, there was a trend for P to be least effective. At the 2% concentrations for the individual metabolites, L was most effective at retarding MMb formation in the Semitendinosus (M was intermediate and P was least effective), and M was most effective in the Psoas major and L. lumborum muscles (L was intermediate and P was least effective). Metmyoglobin was reduced most effectively with a combination of metabolites (M + L > M + P > L + P). Enhancing meat with these metabolites can effectively extend colour life of post-rigor meat, apparently by providing more reducing conditions for myoglobin, thus increasing myoglobin redox form stability.

Myoglobin: A scavenger of bioactive NO.

The present study explored the role of myoglobin (Mb) in cardiac NO homeostasis and its functional relevance by employing isolated hearts of wild-type (WT) and myoglobin knockout mice. (1)H NMR spectroscopy was used to measure directly the conversion of oxygenated Mb (MbO(2)) to metmyoglobin (metMb) by reaction with NO. NO was applied intracoronarily (5 nM to 25 microM), or its endogenous production was stimulated with bradykinin (Bk; 10 nM to 2 microM). We found that infusion of authentic NO solutions dose-dependently (>/= 2.5 microM NO) increased metMb formation in WT hearts that was rapidly reversible on cessation of NO infusion. Likewise, Bk-induced release of NO was associated with significant metMb formation in the WT (>/=1 microM Bk). Hearts lacking Mb reacted more sensitively to infused NO in that vasodilatation and the cardiodepressant actions of NO were more pronounced. Similar results were obtained with Bk. The lower sensitivity of WT hearts to changes in NO concentration fits well with the hypothesis that in the presence of Mb, a continuous degradation of NO takes place by reaction of MbO(2) + NO to metMb + NO(3)(-), thereby effectively reducing cytosolic NO concentration. This breakdown protects myocytic cytochromes against transient rises in cytosolic NO. Regeneration of metMb by metMb reductase to Mb and subsequent association with O(2) leads to reformation of MbO(2) available for another NO degradation cycle. Our data indicate that this cycle is crucial in the breakdown of NO and substantially determines the dose-response curve of the NO effects on coronary blood flow and cardiac contractility.

Effect of Pseudomonas fluorescens on beef discoloration and oxymyoglobin oxidation in vitro.

The relationship between bacterial growth and oxymyoglobin oxidation in vitro and in meat was studied. In the in vitro study, oxymyoglobin was combined with Pseudomonas fluorescens or sterile nutrient broth (control) in an airtight vessel. P. fluorescens samples showed greater metmyoglobin formation and oxygen consumption than controls. The P. fluorescens population in the reaction vessels was correlated with metmyoglobin formation (r = 0.85, P < 0.05) and oxygen consumption (r = 0.91, P < 0.05). When P. fluorescens and oxymyoglobin were combined in an airtight vessel, reducing the headspace from 13 ml and 9 ml to 3 ml resulted in greater metmyoglobin formation (P < 0.05). In the meat study, beef cores prepared from longissimus lumborum were inoculated with P. fluorescens (10(7) CFU/cm2) or sterile peptone water (control), packaged under 1% O2 (+99% N2), air, or 100% O2 and stored at 4 degrees C. Inoculated beef cores showed higher bacterial loads and metmyoglobin formation than their respective controls during 10 h storage in 1% O2, 3 days in air, and 7 days in 100% O2 (P < 0.05). This finding indicated that P. fluorescens could accelerate beef discoloration. Overall, studies demonstrated that oxygen consumption concomitant with P. fluorescens growth decreased partial oxygen pressure, which accelerated oxymyoglobin oxidation.

Ferrylmyoglobin formation induced by acute magnesium deficiency in perfused rat heart causes cardiac failure.

The oxidation states of intracellular myoglobin and cytochrome oxidase aa3 were monitored by reflectance spectrophotometry in isolated perfused rat hearts subjected to an acutely magnesium deficient environment. After exposure to low extracellular [Mg2+]o (i.e., 0.3 mM) for 30 min, more than 80% of the oxymyoglobin converted to its deoxygenated form. The level of reduced cytochrome oxidase aa3 also increased about 80% in low [Mg2+]o. The deoxymyoglobin was converted further to a species identified as ferrylmyoglobin by its reaction with Na2S to form ferrous sulfmyoglobin which was optically visible. This process, set into motion by acute Mg deficiency, resulted from a direct accessibility of the exogenous peroxide to the cytosolic protein. The results suggest that a pathway leading to cardiac tissue damage, induced by magnesium deficiency, is probably involved in the generation of a ferrylmyoglobin radical which could be prevented by addition of ascorbate, which is known to be a one-electron reductant of this hypervalent form of myoglobin. In further studies, we also investigated whether addition of different concentrations of ascorbic acid (AA) to the perfusate could enhance myocardial function after exposure to low [Mg2+]o perfusion. Four concentrations of AA (0.5, 1, 5, 10 mM) were tested, and the results indicate that they exert their effects in a concentration-dependent manner; 1 mM AA was the most effective dose in improving aortic output in a Mg-deficient heart. Ferrylmyoglobin formation was found to be formed considerably before intracellular release of either creatine phosphokinase or lactic dehydrogenase. These studies may have wide implications as a new mechanism by which low extracellular Mg2+ can induce myocardial injury and subsequent cardiac failure.

Tissue equilibration and subcellular distribution of vitamin E relative to myoglobin and lipid oxidation in displayed beef.

Supplementary alpha-tocopheryl acetate (vitamin E) was fed to provide none (E0), 2,000 IU/d (E2000), 5.8 IU/kg live weight (E5.8), or 8.6 IU/kg live weight (E8.6) to steers that were individually fed mainly a corn diet. Three steers were placed on each of 10 treatments: E0, E2000, E5.8, E5.8 to d 126 then E0 to d 266, E0 to d 126 then E5.8 to d 266, E8.6, grazing followed by either E0 or E8.6 all with Holstein steers; and E0 and E2000 with crossbred beef steers. During the last 100 d, vitamin E consumption (International Units/day) averaged 96 for E0, 1,840 for E2000, 2,520 for E5.8, and 3,610 for E8.6. Concentrations of alpha-tocopherol in plasma and in liver and longissimus lumborum biopsy samples obtained every 42 d were elevated (P < .01) by vitamin E supplementation. Tissue saturation was approached at these vitamin E intakes causing similar incorporation of alpha-tocopherol with both per day and per BW supplementation strategies. Maximum accretion or depletion of alpha-tocopherol in plasma and liver occurred before 42 d, but accretion required 120 d and depletion required 180 d in longissimus lumborum. Vitamin E supplementation elevated (P < .01) concentrations of alpha-tocopherol in liver, lung, subcutaneous fat, omental fat, perirenal fat, kidney, diaphragm, spinal cord, longissimus lumborum, and plasma at slaughter with maximum accretion achieved (P < .01) in lung, subcutaneous fat, kidney, diaphragm, and spinal cord. Depletion was not achieved in longissimus lumborum and spinal cord (P < .01), subcutaneous fat (P < .06), and perirenal fat (P < .08) within 140 d. Vitamin E inhibited (P < .01) oxidation at the surface and center of longissimus lumborum steaks displayed for 19 d. Lipid oxidation occurred throughout E0 steaks, but metmyoglobin accumulation occurred more rapidly (P < .01) on the surface than in the center. Myoglobin and lipid oxidation were not concurrent events. Supplementation with vitamin E increased (P < .01) alpha-tocopherol concentrations in longissimus lumborum fractions (mitochondria, microsome, cytoplasm, connective, and remainder) but, except for connective tissue, the proportional distribution of total longissimus lumborum alpha-tocopherol was not affected (P > .1) by vitamin E supplementation. Vitamin E supplementation for at least 44 d at 1,300 IU/d is expected to incorporate adequate amounts of alpha-tocopherol into muscle (3.3 micrograms/g for longissimus lumborum) to produce beef with extended color and lipid stability.

Oxidation of cardiac myoglobin in vivo by sodium nitrite or hydroxylamine.

A non-vascularized fish heart model was used to assess the oxidation of cardiac myoglobin in vivo by compounds known to cause methemoglobinemia. Buffalo sculpin (Enophrys bison) were cannulated from the afferent branchial artery to permit repeated blood sampling and injected intraperitoneally with sodium nitrite, hydroxylamine or aniline. Methemoglobin was formed by sublethal levels of sodium nitrite or hydroxylamine. For hydroxylamine, the time to peak effect was less than 1 h. For sodium nitrite, the onset was less rapid and the effect more prolonged. Aniline had no effect on hemoglobin at any concentration tested. Cardiac myoglobin, assayed at the time of peak effect on hemoglobin, was oxidized in a dose-dependent manner by sodium nitrite or hydroxylamine. At high doses of sodium nitrite (50 and 100 mg/kg), the oxidation of myoglobin exceeded that of hemoglobin. The reverse was true of hydroxylamine at all concentrations tested. This study suggests that possibility that cardiac myoglobin is oxidized in occupational or other exposures to sodium nitrite, hydroxylamine and related compounds.