Mechanism of oxymyoglobin oxidation in the presence of oxidizing lipids in bovine muscle

Lipid oxidation and oxymyoglobin oxidation were measured in bovine muscle homogenates. M. longissimus dorsi homogenates (25% w/w, pH 5.7) were prepared, held at 4 degrees C, and subjected to one of the following treatments: (i) stirred and bubbled with oxygen, (ii) stirred with no oxygen, or (iii) neither stirred nor bubbled with oxygen (control). Lipid oxidation was initiated with 45 microM ferric chloride/sodium ascorbate. Lipid oxidation was highest and oxymyoglobin oxidation lowest in the homogenate bubbled with oxygen while lipid oxidation was lowest and oxymyoglobin oxidation highest in the control homogenate. Dissolved oxygen became depleted over time in the control homogenate, remained high in the homogenate bubbled with oxygen, but decreased and then increased in the homogenate stirred with no oxygen. Free radical formation was lower in the control homogenate than in the stirred homogenates as determined by spin trapping and electron spin resonance detection. The data indicated that lipid oxidation-induced oxygen depletion, as opposed to primary or secondary lipid oxidation products, is a likely cause of oxymyoglobin oxidation in muscle systems.     

Electron spin resonance spin trapping for analysis of lipid oxidation in oils: inhibiting effect of the spin trap alpha-phenyl-N-tert-butylnitrone on lipid oxidation

The electron spin resonance (ESR) spin trapping technique was investigated as an analytical approach to follow lipid oxidation of rapeseed oil, sunflower oil, and fish oil during storage at 40 degrees C. Unlike previous investigations, alpha-phenyl-N-tert-butylnitrone (PBN), used as spin trap, was added to the fresh oils and formation of radicals was monitored during storage. Results were compared with the development in peroxide value (PV) and the thiobarbituric acid index (TBA). Increasing radical development was detected during the initial stages of oxidation, during which no significant changes in PV and TBA were observed. Evidence of spin adduct depletion was found during prolonged storage, suggesting that although spin trapping of radicals may be used to follow early events in lipid oxidation, it is not a suitable parameter for long periods of time. Addition of the spin trap after sequential samplings is recommended for getting an insight of oxidative changes during storage. Further, the influence of the spin trap (PBN) on lipid oxidation was studied in detail by application of PV and TBA and by following the depletion of naturally occurring tocopherol. PBN was found to possess a profound inhibiting effect on lipid oxidation. Such an effect was found to be dependent on the nature of the oil, and it was observed that the lower the oxidative stability, the larger the effect of PBN on lipid oxidation. This effect was interpreted in terms of the capability of PBN to react with peroxyl radicals, which in turn depends on the initial tocopherol content of the oils.     

β-carotene radical cation addition to green tea polyphenols. Mechanism of antioxidant antagonism in peroxidizing liposomes

Green tea polyphenols, (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG), all showed antioxidative effect in liposomes for lipid oxidation initiated in the lipid phase (antioxidant efficiency EC > EGCG > ECG > EGC) or in the aqueous phase (EC ? EGC > EGCG > ECG) as monitored by the formation of conjugated dienes. For initiation in the lipid phase, β-carotene, itself active as an antioxidant, showed antagonism with the polyphenols (EC > ECG > EGCG > EGC). The Trolox equivalent antioxidant capacity (TEAC EGC > EGCG > ECG > EC) correlates with the lowest phenol O-H bond dissociation enthalpy (BDE) as calculated by density functional theory (DFT). Surface-enhanced Raman spectroscopy (SERS) was used to assess the reducing power of the phenolic hydroxyls in corroboration with DFT calculations. For homogeneous (1:9 v/v methanol/chloroform) solution, the β-carotene radical cation reacted readily with each of the polyphenol monoanions (but not with the neutral polyphenols) with a rate approaching the diffusion limit for EC as studied by laser flash photolysis at 25 °C monitoring the radical cation at 950 nm. The rate constant did not correlate with polyphenol HOMO/LUMO energy gap (DFT calculations), and β-carotene was not regenerated by an electron transfer reaction (monitored at 500 nm). It is suggested that the β-carotene radical cation is rather reacting with the tea polyphenols through addition, as further evidenced by steady-state absorption spectroscopy and liquid chromatography-mass spectroscopy (LC-MS), in effect preventing regeneration of β-carotene as an active lipid phase antioxidant and leading to the observed antagonism.     

Method for the estimation of nutrient uptake kinetic parameters using nonparametric statistics

The uptake of many plant nutrients has been shown to follow Michaelis‐Menten enzyme kinetics, and as a result several methods of collecting and analyzing uptake data have been developed. The method proposed here consists of a continuously flowing hydroponic system and a method of data analysis that estimates a value of K_mand V_max for each plant. The method is nondestructive; it does not require large amounts of space; and the plants are near steady‐state uptake. In simulated experiments with various assumptions about variability in the data, the nonparametric statistical analysis of the results provided estimates as good as or better than regression analysis estimates of the two parameters of the Michaelis‐Menten equation.     

Is humane slaughter of fish possible for industry?

Abstract The objective was to evaluate industrial and research slaughter methods for Atlantic salmon (Salmo salar), gilt‐head seabream (Sparus auratus) and eel (Anguilla anguilla) with respect to welfare and quality. As a general term of reference, an optimal slaughter method should render fish unconscious until death without avoidable excitement, pain or suffering prior to killing. For Atlantic salmon, commercial slaughter methods (carbon dioxide stunning followed by gill cutting, and gill cutting alone) are not in conformity with the general term of reference, as the fish are not rendered unconscious immediately and possibly experience stress. Evaluation of automated percussive stunning remained unconclusive. More research should enable us to ascertain whether loss of consciousness is instantaneous. Electrical stunning can be humane if applied properly. However, because flesh of electrostunned fish was characterized by occasional bloodspots, optimization of the electrical parameters is required. Prototypes for percussive and electrical stunning of salmon have been recently developed. This implies that humane slaughter of salmon is feasible for industry. For gilt‐head seabream, neither aphyxia in air nor transfer of the fish to an ice slurry were considered to be humane: the methods did not induce immediate brain dysfunction and vigorous attempts to escape occurred. Percussive and electrical stunning can be in conformity with the general term of reference. However, conditions for stunning whole batches of seabream have not been established. Quality of the fish slaughtered by percussive stunning was similar to that obtained by the industrial method, i.e. immersion in an ice slurry. Further work is required to establish optimal stunning conditions and to develop prototypes. For eel, desliming in a salt‐bath followed by evisceration, electrical stunning performed under the conditions prescribed by the German legislation, and live chilling and freezing were not considered to be humane. In contrast, it was established that a 10–20 kg batch of eels in fresh water could be rendered unconscious immediately and until death by applying electricity in combination with nitrogen gas. The conditions used were 0.64 A dm^?2 for 1 s, followed by 0.17 A dm^?2 combined with nitrogen flushing for 5 min. A preliminary assessment of flesh quality suggests that it may be improved by application of the latter method, compared with the salt bath. The results clearly indicated that humane slaughter of eels is possible in practice.     

Use of Water for Swimming and its Relationship to Temperature and Other Factors in Farm Mink (Mustela vison)

An investigation into swimming behaviour and its relationship to outdoor temperature was conducted in 40 ranch mink. Two different cage sizes were used and all animals had access to a basin filled with water 15 cm deep. During the years 1994-1998, water loss from the basin and temperatures at a nearby weather station were recorded once a week. In the last year 11 mink were video-recorded for 24 h. Using the monthly average daily water loss as an indicator, swimming activity showed a clear annual variation over 4 years and was positively correlated with average monthly temperature. With regard to swimming activity, a large individual variation existed: average water loss varied from 10 ml to nearly 4 litres per day, with those in large cage units having the greatest water loss. The video-recordings confirmed this variation: the number of swims ranged from zero to 177 during 24 h and there was an indication of more swimming bouts in the mink in the large cage units. The duration of swims varied from 2 to 55 s per bout. The relationship between swimming and general activity is discussed, along with the importance of water for swimming as a means of thermoregulation.     

VALIDATION OF DELAYED GADOLINIUM‐ENHANCED MAGNETIC RESONANCE IMAGING OF CARTILAGE AND T2 MAPPING FOR QUANTIFYING DISTAL METACARPUS/METATARSUS CARTILAGE THICKNESS IN THOROUGHBRED RACEHORSES

The purpose of this study was to determine whether delayed gadolinium‐enhanced magnetic resonance imaging of cartilage (dGEMRIC) and T2 mapping are accurate techniques for measuring cartilage thickness in the metacarpus3/metatarsus3 (Mc3/Mt3) of Thoroughbred racehorses. Twenty‐four Mc3/Mt3 cadaver specimens were acquired from six healthy racehorses. Cartilage thickness was measured from postintra‐articular Gd‐DTPA^2? images acquired using short tau inversion recovery (STIR), and proton density weighted (PDw) sequences, and compared with cartilage thickness measured from corresponding histologic images. Two observers performed each histologic measurement twice at three different sites, with measurement times spaced at least 5 days apart. Histologic cartilage thickness was measured at each of the three sites from the articular surface to the bone–cartilage interface, and from the articular surface to the mineralized cartilage interface (tidemark ) . Intra‐observer repeatability was good to moderate for dGEMRIC where Mc3/Mt3 cartilage was not in contact with the proximal phalanx. Where the Mc3/Mt3 cartilage was in contact with the proximal phalanx cartilage, dGEMRIC STIR and T2 mapping PDw cartilage thicknesses of Mc3/Mt3 could not be measured reliably. When measured from the articular surface to the bone–cartilage interface, histologic cartilage thickness did not differ from STIR or PDw cartilage thickness at the site where the Mc3/Mt3 cartilage surface was separated from the proximal phalanx cartilage (P > 0.05). Findings indicated that dGEMRIC STIR and T2 mapping PDw are accurate techniques for measuring Mc3/Mt3 cartilage thickness at locations where the cartilage is not in direct contact with the proximal phalanx cartilage.     

Antioxidants and shelf life of whole wheat bread

Industrially produced bread normally operates with a shelf life of several weeks at room temperature and indications of storage-related off-flavour development as a consequence of lipid oxidation have been suggested. The present study has tested enrichment of whole wheat bread with α-tocopherol or commercially used rosemary extracts for production of bread loaves with higher oxidative stability and hence better overall sensory quality. Bread quality was evaluated by sensory profiling, determination of antioxidative capacity, determination of lipid hydroperoxides as primary oxidation products, and analysis of volatile compounds including secondary lipid oxidation products. Enrichment of bread with α-tocopherol resulted in higher degrees of rancid aroma and flavour in fresh samples, which was explained by higher levels of secondary oxidation products, whereas enrichment of bread with rosemary extracts did not have any effect. Accordingly, application of antioxidants cannot, based on the current data, be recommended for achievement of bread with improved sensory properties during storage.