A preliminary study on the changes in some potential markers of muscle-cell degradation in sub-maximally exercised horses supplemented with a protein and amino acid mixture.

In this preliminary study, time-dependent changes in plasma CK and AST activity, tyrosine (Tyr), 3-methyl-histidine (3mHis), glucose and lactate concentrations were analysed in nine horses under two different conditions. Furthermore, intramuscular concentrations of Tyr, 3mHis and activities of cathepsin B, acid phosphatase (ACP), glucose-6-phosphate dehydrogenase (G6PDH) and mRNA expression of ubiquitin were determined at the same time. After studying the effects of exercise alone, the effects of exercise and feeding of an experimental protein/amino acid (AA) supplement were analysed. Horses were submitted to a total of four standardised exercise tests (SETs) of high intensity. Potential markers of muscle break down were determined prior to, immediately after, 4 and 18 h after exercise. The experiment was subdivided into two consecutive periods of 3 weeks. In each period, two SETs were performed. In the second period, horses were fed with the protein/AA supplement within 1 h after exercise. Significant changes in plasma, intramuscular Tyr levels and mRNA expression of ubiquitin were caused both by time in relation to exercise and by treatment with the protein/AA supplement. The experimental supplement significantly decreased the 4-h post-exercise expression of ubiquitin mRNA in muscle. Only a borderline increase of markers of lysosomal involvement was seen and CK and AST activity generally showed their normal post-exercise patterns. A clear post-exercise reduction of this CK activity, however, was not observed after supplementation with the protein/AA mixture. The current findings indicate that horses might benefit from protein and AA supplementation directly after training by decreasing post-exercise proteolysis. The results support that further studies should be performed to characterize changes in equine protein metabolism caused by exercise including underlying molecular mechanisms.

Changes in intramuscular amino acid levels in submaximally exercised horses - a pilot study.

The time-dependent changes in intramuscular amino acid (AA) levels caused by exercise and by feeding a protein/AA supplement were analysed in nine horses. Horses were submitted to a total of four standardized exercise tests (SETs). Amino acid concentrations were determined prior to, immediately after, 4 and 18 h after exercise. The experiment was subdivided into two consecutive periods of 3 weeks. In each period two SETs were performed. In the second period, horses were given a protein/AA supplement within 1 h after exercise. Significant changes in mean plasma AA levels similar to previous studies were noted to be time-dependent and to be associated with feeding the supplement. The intramuscular concentrations of the free AA in relation to pre-exercise levels showed significant time-dependent changes for alanine, asparagine, aspartate, citrulline, glutamine, glycine, isoleucine, leucine, methionine, serine, taurine, threonine, tyrosine and valine. Feeding the supplement significantly increased the 4 h post-exercise intramuscular concentration of alanine, isoleucine, methionine and tyrosine. At 18 h after exercise, apart from isoleucine and methionine, levels were still increased and also those of asparagine, histidine and valine in relation to none treatment. Hence, it was concluded that AA mixtures administered orally to horses within 1 h after exercise increased intramuscular AA pool.

The effects of short intensive exercise on plasma free amino acids in standardbred trotters.

The aim of this study was to investigate the effect of short intense exercise on plasma amino acid concentrations in trotters and to test the repeatability of plasma amino acids concentration in samples obtained on two independent days under field conditions. Plasma amino acid concentrations were analysed in blood samples of 36 standardbred trotters before and after intense exercise over a distance of 2000 m. Sampling was repeated in 20 horses after 35 days. Exercise intensity was estimated from post-exercise lactate levels. Horses were divided in two groups according to a cut-off lactate concentration at 15 mmol/l. The plasma concentrations of alanine, aspartate, glutamate, isoleucine, leucine, lysine and taurine increased and arginine, asparagine, citrulline, glutamine, glycine, histidine, methionine, serine, tryptophan and 3-methylhistidine decreased after exercise. Ornithine, threonine, tyrosine, phenylalanine and valine concentrations remained constant. Higher intensity of exercise significantly decreased tryptophan and increased taurine concentrations. Sampling day had a significant effect on the absolute pre- and post-exercise amino acid concentrations. Exercise had a significant influence on the concentrations of most plasma amino acids in trotters. These changes could reflect shifts between the free amino acid compartments, but there were also some indications for muscle catabolism. The amino acid supply of sporting horses could be of specific significance for maintaining muscle integrity and for the improvement of post-exercise recovery of competition horses.

Individual differences and repeatability of post-prandial changes of plasma-free amino acids in young horses.

Few data are available on post-prandial changes of plasma amino acids (AAs) in horses and on the repeatability and the individual variance on different sampling days. The objective of the present study was to measure pre- and post-prandial concentrations of plasma AA in 10 yearling horses. Blood samples were taken on days 1 and 40 of the study before feeding of hay, oats and soya meal and over an 8 h post-prandial period in 2-h intervals. The plasma AAs were measured by high-pressure liquid chromatography after ortho-phthalaldehyde derivatization. Mean fasting concentrations of the AAs were not significantly influenced by the individuum and sampling day. Repeatability of the fasting AA levels in the individual horses on two different sampling days was only found for histidine, 3-methylhistidine, methionine, tryptophan and taurine. While the absolute post-prandial AA concentrations differed between sampling days, the relative changes were comparable. All AA concentrations except 3-methylhistidine increased after feeding by 13% to more than 200% of their fasting values if the combined data of both days were analysed. Four hours after feeding the concentrations of arginine, asparagine, lysine, leucine, isoleucine and threonine, decreased more than 20%. Histidine, methionine, phenylalanine, valine, tryptophan, glutamine, glycine, tyrosine and taurine concentrations decreased by less than 20%. Concentrations of aspartic acid, glutamic acid, ornithine, serine and citrulline remained elevated. Most AA approached the fasting concentrations at 8 h, only glycine increased between 6 and 8 h after meal and 3-methyl-histidine concentrations were constant throughout the entire period. In conclusion, the pre-prandial plasma AA in horses appeared less influenced by individuum or sampling day than post-prandial plasma AA concentrations. Therefore, plasma AA concentrations should be interpreted only under well-defined conditions, especially regarding the feeding regimen.

Detection of oestrogen and progesterone receptor expression in breast tumors by semiquantitative PCR.

BACKGROUND: Estimation of oestrogen receptors (ER) and progesterone receptors (PgR) by dextran-coated charcoal (DCC) or immunohistochemical methods have become standard practices in the management of breast cancer. MATERIALS AND METHODS: A "multiplex" polymerase chain reaction (PCR) system was developed for quantitative estimation of ER and PgR mRNA in breast tumour specimens. RESULTS: A statistically significant correlation could be found between the mRNA of the oestrogen and the progesterone receptor (p < or = 0.0001). Protein data defined in classes, compared with mRNA data showed a significant correlation for the oestrogen receptor (p < or = 0.0001) as well as for the progesterone receptor (p < or = 0.046). CONCLUSION: Messenger RNA could be determined by the present PCR system in tumours assayed as negative by DCC method. Therefore, this sensitive PCR procedure, which requires small amounts of material may be very useful as a diagnostic test to determine the choice of therapy.