Effects of different storage conditions on quality related porphyrin fluorescence signatures of pork slices.

This study evaluated the potential of fluorescence as an indicator of pork quality by determining the effects of various conditions on fluorescence signatures (excitation at 420 nm, emission at 550-750 nm). Storage of porcine musculus longissimus dorsi in PE bags led to a clear increase in porphyrin fluorescence intensity after approx. 10 d post mortem. Modified gas atmosphere (70% O(2), 30% CO(2)) inhibited the fluorescence emission of zinc protoporphyrin and protoporphyrin IX due to quenching by oxygen. Bleaching processes caused similar effects by halogen light exposure during meat storage. However, already formed signals could not be manipulated by oxygen or halogen light. Storage under vacuum reduced the quenching effects and resulted in increased fluorescence intensities. Freezing and thawing of meat samples delayed and reduced the increase in fluorescence intensity. Only minor effects could be detected at long-term frozen storage for two months. Consequently porphyrin fluorescence analysis is a potential means to indicate changes of pork quality and remaining shelf life.

Chemical safety of meat and meat products.

Since the Second World War the consumer behaviour in developed countries changed drastically. Primarily there existed the demand for sufficient food after a period of starvation, afterwards the desire for higher quality was arising, whereas today most people ask for safe and healthy food with high quality. Therefore a united approach comprising consistent standards, sound science and robust controls is required to ensure consumers' health and to maintain consumers' confidence and satisfaction. Chemical analysis along the whole food chain downstream (tracking) from primary production to the consumer and upstream (tracing) from the consumer to primary production is an important prerequisite to ensure food safety and quality. In this frame the focus of the following paper is the "chemical safety of meat and meat products" taking into account inorganic as well as organic residues and contaminants, the use of nitrite in meat products, the incidence of veterinary drugs, as well as a Failure Mode and Effect Analysis (FMEA) system assessing (prioritizing) vulnerable food chain steps to decrease or eliminate vulnerability.

Fluorimetric detection of protoporphyrins as an indicator for quality monitoring of fresh intact pork meat.

In fresh meat production fast and non-destructive quality monitoring along the distribution chain is a key aspect to guaranteeing high quality and safe products for consumption. The applicability of fluorescence spectroscopy using protoporphyrins as indicators for meat ageing was investigated. Porcine musculus longissimus dorsi (MLD) was stored in slices over 20 days at 5 and 12°C and measured every day with an excitation of 420nm and an emission range of 550-750nm. Additionally, pH, drip loss and colour were examined to assess possible correlations. The obtained spectra of the MLD showed an increase in three peaks at 592, 638 and 705nm which could be reconstructed using the spectra of standard solutions of protoporphyrin IX (PP) and zinc protoporphyrin IX (ZnPP) or magnesium protoporphyrin (MgPP), respectively. Using principal component analysis (PCA) on the fluorescence spectral data, the meat slices stored at 5°C showed differences in the fluorescence signal after the 10th day and 5th day when stored at 12°C. An interrelationship between the additional analyses and the fluorescence intensities on these relevant days could not be established. In conclusion, the increase of ZnPP fluorescence due to temperature related changes of physiological meat properties is capable of serving as a quality indicator with regards to inadequate conditioning (e.g. during transportation and/or storage) of pork meat.

Albumen freshness assessment by combining visible near-infrared transmission and low-resolution proton nuclear magnetic resonance spectroscopy.

Recently, some nondestructive methods for the assessment of albumen freshness were developed. Among others, visible near-infrared transmission spectroscopy and low-resolution proton nuclear magnetic resonance (LR (1)H NMR) measurements were proposed. This study was performed to evaluate the potential of the combined measurement of visible near-infrared transmission spectroscopy and LR (1)H NMR measurements for the assessment of albumen freshness. Our results show that solely based on the transmission measurements, a good estimation of albumen freshness can be achieved. Based on LR (1)H NMR measurements, an estimation of albumen freshness can be achieved if larger egg collectives are used. However, when individual eggs are considered, only a moderate estimation is feasible. Finally, it was observed that combining both spectroscopic techniques did not improve the assessment of albumen freshness when compared solely to transmission measurements.

Traceability from a European perspective.

At pan-European level there is a need for traceability systems giving information on origin, processing, retailing and final destination of foodstuffs. Such systems shall enhance consumer confidence in food; enable the regulatory authorities to identify and to withdraw health hazardous and non-consumable foodstuffs from the market. Animal feeds are an element in this "food-to-farm" approach to public health. Such feedstuffs are preliminary elements of some foods for human consumption, and hence are an inherent element of the food chain. A harmonised pan-European food traceability protocol would greatly assist authorities in detecting fraud as well as dangerous substances. The food chain comprises a range of sequential and parallel stages bridging the full spectrum from agricultural production to the consumable foodstuffs by consumers. EU legislation on traceability and the technologies needed to implement this system for meat and meat products are the focus of this paper.

Comparative studies of pyruvate kinase from PSE and normal pig muscles.

A fast breakdown of glycogen is observed in muscles of stress-susceptible pigs leading to pale, soft and exudative (PSE) meat. We report a comparative study of pyruvate kinase from muscles of normal and PSE-prone pigs. Compared with the enzyme from normal muscle, pyruvate kinase isolated from PSE muscle shows a five times lower Michaelis constant, Km, for phosphoenol pyruvate and a more than ten times higher Kcat/Km value. The pH dependency of the enzymatic activity is shifted to more acidic values for pyruvate kinase from PSE muscles. According to isoelectric focusing, pyruvate kinase from PSE muscle consists of three isoforms, while only two isoforms are detectable in pyruvate kinase preparations from normal pigs. The various isoforms were isolated by preparative isoelectric focusing and their steady-state properties were compared. Isoform 3, which is found only in PSE muscle, shows a 10-fold higher specific activity, a 30-fold lower Km value and a 100-fold increased kcat/Km value for phosphoenol pyruvate as compared to isoform 1. The presence of isoform 3 in PSE muscle appears to be responsible for the high activity of this enzyme under the more acidic conditions prevailing in PSE muscle. In vitro phosphorylation and dephosphorylation experiments using total enzyme and purified isoenzyme 1 suggest that isoforms 2 and 3 arise from isoform 1 by phosphorylation. Thus protein phosphorylation seems to be responsible for the shift in activity of pyruvate kinase, a key enzyme of glycolysis, under the acidic conditions of PSE muscles.

Enzymological investigations on the causes for the PSE-syndrome, I. Comparative studies on pyruvate kinase from PSE- and normal pig muscles.

A fast breakdown of glycogen is observed in muscles of stress-susceptible pigs leading to pale, soft and exudative (PSE) meat. We report a comparative study of pyruvate kinase from muscles of normal and PSE-prone pigs. Compared with enzyme from normal muscle, pyruvate kinase isolated from PSE-muscle shows a five times lower K(m) for phosphoenol pyruvate and a more than ten times higher k (cat)K (m) value. The pH-dependency of the enzymatic activity is shifted to more acidic values for pyruvate kinase from PSE muscles. According to isoelectric focusing, pyruvate kinase from PSE muscle consists of three isoforms, while only two isoforms are detectable in pyruvate kinase preparations from normal pigs. The various isoforms were isolated by preparative isoelectric focusing and their steady-state properties were compared. Isoform 3, which is found only in PSE muscle, shows a 10-fold higher specific activity, a 30-fold lower K(m) value and a 100-fold increased k (cat)K (m) value for phosphoenol pyruvate compared to isoform 1. The presence of isoform 3 in PSE-muscle appears to be responsible for the high activity of this enzyme under the more acidic conditions prevailing in PSE-muscle. In vitro phosphorylation and dephosphorylation experiments using total enzyme and purified isoenzyme 1 suggest that isoforms 2 and 3 arise from isoform 1 by phosphorylation. Thus protein phosphorylation seems to be responsible for the shift in activity of pyruvate kinase, a key enzyme of glycolysis, under the acidic conditions of PSE-muscles.

Enzymological investigations on the causes for the PSE-syndrome, II. Comparative studies on glycogen phosphorylase from pig muscles.

In order to investigate the cell biological causes for the fast breakdown of glycogen which is observed during the development of the PSE (pale, soft, exudative) syndrome in muscles of stress-susceptible pigs, muscle glycogen phosphorylase (GP) as a key enzyme in two isoforms, a and b, of the energy turnover was isolated from M. longissimus dorsi of normal and PSE-prone pigs of the German Landrace. GP b as well as GP a from normal and PSE-muscles exist in a dimeric form with a molecular weight of 97 000 D per subunit. The tendency for tetramerization of GP b increases in the presence of ATP, whereas the enzyme activity is simultaneously inhibited. The catalytic activities of GP a and GP b from both groups of animals show an optimum at pH 7.0. GP b can be activated to GP a by phosphorylation with the result of a 25% higher optimum specific activity in the case of normal and PSE-muscles. In interaction with glycogen and glucose-1-phosphate GP b follows the characteristics of a Michaelis-Menten kinetic, whereas the binding of AMP and phosphate proves to be allosteric. In comparison of the structural and kinetic characteristics of GP from normal as well as PSE-muscles no significant differences could be determined, indicating that GP does not belong to those factors which are triggering an accelerated energy turnover of ATP in muscles of stress-susceptible pigs.

Purification and isoenzymic composition of glycogen phosphorylase b from normal and abnormal (PSE) muscles.

Ion exchange chromatography and preparative isoelectric focusing allowed the identification of five isoenzymes of glycogen phosphorylase b from the longissimus dorsi muscle of normal pigs and those prone to having pale, soft and exudative (PSE) muscle. The isoelectric point of the isoenzymes varied in the pH range from 6.29 to 6.55. One of them, with an isoelectric point at about a pH of 6.49, accounts for 65% of the total glycogen phosphorylase b activity. No significant differences between normal and PSE-prone pigs were observed in the total glycogen phosphorylase b activity and in the isoenzyme distribution pattern. It is concluded that the fast glycogen turnover in PSE-prone pigs is not due to a different isoenzyme pattern of phosphorylase b.

[Myoglobin content and beta-hydroxyacyl-CoA-dehydrogenase activity of different muscles of cattle and calf]. / Myoglobingehalt und beta-hydroxyacyl-CoA-Dehydrogenase-Aktivität verschiedener Muskeln von Rind und Kalb.

The heart, tongue, jowl, diaphragm and tail as well as shoulder, top round, the longissimus dorsi muscle of slaughtered cattle and the diaphragms of calf were examined with respect to their myoglobin content and beta-hydroxyacyl-CoA-dehydrogenase (HADH) activity. According to Gottesmann and Hamm [1] the product of these two values, the so-called MH value, can serve as the differentiation between the diaphragm and "normal cross striated skeletal muscles". Like the diaphragm, heart, tongue and jowl of cattle show higher MH values than those of "normal beef". Muscles in the tail have the same MH values as those of normal beef muscles. There are no essential differences in the MH values of various cross-striated muscle types of cows and calves. Muscles of cattle show a slightly higher myoglobin content, whereas the HADH activity is lower than in veal.

Specific recognition of the 3'-terminal adenosine of tRNAPhe in the exit site of Escherichia coli ribosomes.

Ribosomes from Escherichia coli possess, in addition to A and P sites, a third tRNA binding site, which according to its presumed function in tRNA release during translocation has been termed the exit site. The exit site exhibits a remarkable specificity for deacylated tRNA; charged tRNA, e.g. N-AcPhe-tRNAPhe, is not bound significantly. To determine the molecular basis of this discrimination, we have measured the exit site binding affinities of a number of derivatives of tRNAPhe from E. coli, modified at the 3' end. Binding to the exit site of the tRNAPhe derivatives was measured fluorimetrically by competition with a fluorescent tRNAPhe derivative. We show here that removal of the 2' and 3' hydroxyl groups of the 3'-terminal adenosine decreases the affinity of tRNAPhe for the exit site 15 and 40-fold, respectively. Substitutions at the 3' hydroxyl group (aminoacylation, phosphorylation, cytidylation) as well as removal of the 3'-terminal adenosine (or adenylate) of tRNAPhe lower the affinity below the detection limit of 2 x 10(5) M-1, i.e. more than 100-fold. Modification of the adenine moiety (1,N6-etheno adenine) or replacement of it with other bases (cytosine, guanine) has the same dramatic effect. In contrast, the binding to both P and A sites is virtually unaffected by all of the modifications tested. These results suggest that a major fraction (at least -12 kJ/mol, probably about -17 kJ/mol) of the free energy of exit site binding of tRNAPhe (-42 kJ/mol at 20 mM-Mg2+) is contributed by the binding of the 3'-terminal adenine to the ribosome. The binding most likely entails the formation of hydrogen bonds.