Determinants of tenderisation in beef Longissimus dorsi and Triceps brachii muscles.

Tenderisation of bovine Mm. longissimus dorsi and triceps brachii and factors impacting tenderisation were studied. Mm. longissimus dorsi and triceps brachii of 12 Friesian-Holstein cows (age 3-11 years; 212-349 kg carcass weight) were sampled at various times post mortem (p.m.) for determination of pH, temperature, fibre type and morphology, connective tissue distribution, SDS-PAGE of myofibrillar proteins, Warner-Bratzler shear force, sarcomere length and osmolality. The stretched position of the M. triceps brachii (sarcomere length 2.35 ± 0.24 µm) resulted in a relatively low shear force at 1 day p.m. (6.2 ± 0.9 kg/cm(2)) with further storage having little additional effect. M. longissimus dorsi entered rigor in a more contracted state (sarcomere length 1.65 ± 0.11 µm), resulting in a relatively high shear force at 1 day p.m. (10.3 ± 2.3). Stepwise linear regression was used to calculate the best 1- to 3-variable equations for shear force of M. longissimus dorsi at 1, 7 and 14 days p.m. and the decrease in shear force between 7 and 14 days p.m. Shear force at 1 day p.m. appeared to be determined mainly by the speed of pH- and temperature-decline. Proteolysis of myofibrillar proteins and animal age appeared to be the main determinants for shear force at 1 and 14 days p.m. The average surface area of type I fibres could explain part of the variation in the decrease in shear force between 1 and 14 days p.m.

Effects on meat quality of the use of clenbuterol in veal calves.

The objective of this study was to examine the effects of clenbuterol administration on meat quality traits of veal. Sixteen Holstein-Friesian veal calves (male) were randomly assigned to one of four treatment groups; either control (n = 4) or clenbuterol-treated (1.6 micrograms.kg [corrected] BW-1.d-1, 42 d) with a withdrawal period between clenbuterol treatment and slaughter of 8 d (n = 4), 4 d (n = 4), or 2 d (n = 4). All animals were slaughtered at the same day at a commercial slaughterplant. At 30 min postmortem the carcasses were split and the right carcass side was electrically stimulated. After 24 h of cooling the longissimus, semimembranosus, triceps brachii, and psoas major muscles were excised and vacuum-packaged. After 1, 7, and 13 d of vacuum storage at 2 +/- 2 degrees C the muscles were sampled to determine tenderness, water-holding capacity, and color characteristics. Clenbuterol treatment resulted in a slower rate of pH decline in the unstimulated longissimus muscle but did not affect the ultimate pH. Clenbuterol treatment resulted in toughening of the longissimus, semimembranosus, and triceps brachii muscles after 1 and(or) 7 d of storage (P < .05). It is suggested that this resulted from a decrease in postmortem proteolysis because both the intensity of a 30-kDa peptide and the myofibril fragmentation index were lower in clenbuterol-treated muscles. Clenbuterol treatment resulted in increased lightness (L*-value) of longissimus and semimembranosus muscles (P < .05), coincident with a lower water-holding capacity. In a following experiment, the effect of clenbuterol administration (0 [n = 5] and 1.0 [n = 5] mg/kg of feed for 27 d) on calpain and calpastatin levels at 1 d postmortem in longissimus muscles of Friesian Pie Noire veal calves was investigated. Clenbuterol administration resulted in an increase in calpastatin levels (P < .05) and a trend (P < 0.1) toward a decrease in mu-calpain activity at 1 d postmortem.

Measurement of totally activated pyruvate dehydrogenase complex activity in human muscle: evaluation of a useful assay.

A sensitive radiochemical method for the determination of the pyruvate dehydrogenase complex (PDHC) activity in skeletal muscle tissue, based on the decarboxylation of [1-14C]-pyruvate to 14CO2, is described. Measurements can be carried out either in muscle homogenate or in 600-g supernatant, both obtainable from a small muscle biopsy specimen (20 mg). In addition to NAD+, thiamine pyrophosphate and coenzyme A in the incubation mixture, a preparation of NADH:cytochrome c reductase (NADHCR) together with cytochrome c has a stimulating effect on the PDHC activity. NADHCR constitutes an oxidation system for NADH to prevent feedback inhibition. Addition of L-carnitine also results in stimulation of PDHC by trapping the produced acetyl-CoA as acetylcarnitine. Special care for radioactive pyruvate, with freeze drying and storage at -20 degrees C under nitrogen, and determination of the purity during every PDHC assay, is required. In the presented assay a Km value of 0.084 mmol/l was found for pyruvate. Nonsigmoidal kinetics was found with a Hill coefficient of 1.63. With the described method, a totally Mg2+,Ca(2+)-stimulated PDHC activity is measured. Addition of a purified specific pyruvate dehydrogenase phosphatase did not yield a higher PDHC activity. Finally, comparison of total PDHC activity with [1-14C]-pyruvate oxidation rates, both measured in the supernatant prepared from fresh muscle, shows an equimolar correlation, indicating that total PDHC activity is rate limiting in the assay for the pyruvate oxidation rate. Neonatal muscle exhibits five to ten times lower PDHC activities and pyruvate oxidation rates than controls (age > 3 years).

The role of ultimate pH in proteolysis and calpain/calpastatin activity in bovine muscle.

Of a total of three Friesian cows, two of which had been treated with adrenalin before slaughter, Mm longissimus (LO), supraspinatus (SS), triceps brachii (TB) and rectus abdominis (RA) were sampled at different times post mortem (pm). pH, calpain/calpastatin activities and degradation of myofibrillar proteins, as evidenced by SDS-PAGE, were assessed. Contraction characteristics were measured by determining myofibrillar ATPase activities. Adrenalin treatment resulted in a high ultimate pH (6.48 +/- 0.40) and a faster decline pm of calpain I activity. The effect was similar in all four investigated muscles (72.4 +/- 5.4% decline at 24 h pm). The decline in calpain I activity in the control muscles was muscle-dependent and ranged from 22.8-74.3% at 24 h pm. Differences in ultimate pH did not lead to distinct rates of breakdown of proteins with molecular weights lower than that of myosin heavy chain. Calpastatin levels were muscle-dependent and correlated with myofibrillar ATPase activity (r = -0.99). In a second experiment Mm rectus abdominis (RA) and psoas major (PM) of adrenalin-treated (n = 6) and control (n = 6) Friesian-Holstein calves were sampled at 1 and 29 h pm for assessment of calpain activities. At seven days pm the M longissimus (LO) was sampled for tenderness evaluation. pH values were measured at 30 min, 4 h and 29 h pm. Adrenalin treatment resulted in a higher ultimate pH in the three muscles. Higher ultimate pH resulted in lower calpain activities in the RA at 29 h pm (P less than or equal to 0.025).(ABSTRACT TRUNCATED AT 250 WORDS)

Estimation of pyruvate dehydrogenase (E1) activity in human skeletal muscle; three cases with E1 deficiency.

Pyruvate dehydrogenase (E1) catalyzes the rate-limiting step of the pyruvate dehydrogenase complex. Since E1 activity of human muscle tissue is low, a sensitive method is needed for diagnostic purposes. Measurement of 14CO2 production from [1-14C]pyruvate provides a specific and sensitive assay for measuring E1 activity. We use as artificial electron acceptor dichlorophenolindophenol (DCPIP) instead of the often applied ferricyanide. The method can be applied to small muscle samples obtained by needle or open biopsy. We prefer to use total homogenate because E1 activities in homogenate are higher than in the corresponding 600-g supernatant of skeletal muscle tissue. Control values in homogenate are higher or of the same order as those reported by others.

Cholesterol metabolism in two strains of rats with high or low response of serum cholesterol to a cholesterol-rich diet.

Transfer from a low cholesterol commercial diet to a high cholesterol diet, containing 2% (wt/wt) cholesterol and 0.5% cholate, caused an increase in serum cholesterol from about 2.5 mmol/L in two inbred rat strains to 5 mmol/L in the hyporesponsive strain and to 20 mmol/L in the hyperresponsive strain. In both strains the excess of cholesterol in the serum was exclusively located in the very low density lipoproteins. Cholesterol feeding caused a sevenfold increase in the amount of cholesterol in the liver, the increase tending to be greater in the hyporesponders. On the commercial diet, the decay of specific radioactivity of serum cholesterol after the intravenous administration of labeled cholesterol was faster in the hyporesponsive rats. The rate of fecal excretion of radioactive bile acids on this diet was higher in the hyporesponders when compared with the hyperresponders, whereas there was no strain difference with regard to the output of fecal neutral steroids. Sterol balance data showed that whole-body cholesterol synthesis on the low cholesterol diet was about twofold higher in the hypo- than in the hyperresponders. When fed the high cholesterol diet the half-life in the serum of injected radioactive cholesterol was about six times shorter in the hyporesponders. In absolute amounts, the hypo- and hyperresponders excreted similar amounts of endogenous (radioactive) bile acids and fecal steroids with the feces on this diet.