Delayed effects of prolonged exercise on serum lipoproteins.

The delayed lipoprotein changes after a 3-hour running test were examined in 14 moderately trained young male subjects. Fasting blood samples were obtained one day before, immediately before, and one, two, and four days after the race. Nonfasting samples were collected immediately after, one, and three hours after exercise. Three hours after the race, the ratio of unesterified cholesterol to cholesteryl esters was significantly increased, and one and two days after the race it was significantly decreased compared to the preexercise value. The HDL2/HDL3 ratio, measured by density-gradient ultracentrifugation was one and three hours after the running significantly elevated. However, no redistribution of the HDL2/HDL3 cholesterol ratio determined by a precipitation method with polyanions was found at this time. One day postexercise HDL cholesterol rose significantly above the preexercise value, and this was associated with an elevation of the HDL3 subfraction. On the following day a significant increase of HDL2 cholesterol and the HDL2/HDL3 cholesterol ratio was found. The apolipoproteins A-I, A-II, and B, measured by radial immunodiffusion, did not change during the first hours and the first two days after the race. On the second postexercise day the Lp(a) lipoprotein rose significantly above the preexercise value. Compared with the preexercise level the LCAT activity was significantly elevated three hours after the race and significantly decreased two days later. The present study suggests that during the first few days after prolonged exercise a number of plasma lipoprotein changes take place that are similar to those observed after a period of physical training.

Can physical exercise induce an effective fibrinolysis?

The study examined serum fibrin(ogen) degradation products (FDP/fdp) after short and prolonged exercise, before and after training, in young and elderly subjects. Six of 15 well-trained soccer players demonstrated an increase of FDP/fdp after a short graded maximal treadmill test. In 12 elderly subjects short maximal exercise produced no increase of FDP/fdp. However, in most of these subjects an exhaustive work load was not reached. Three of 16 moderately trained young subjects exhibited a small FDP/fdp increase after 45 minutes of submaximal exercise and in 3 of 10 participants an elevation of FDP/fdp was found after a 3 h-running test. Immediately after the race there was a significant (p less than 0.001) increase of fragment D, measured by an enzyme immunoassay. During a second 3 h-race after an 8 week physical training program, the post-exercise FDP/fdp and fragment D increase was slightly enhanced. Plasma fibrinogen was significantly (p less than 0.05) elevated on the fourth day after the 3 h-race before but not after the training program. In conclusion intense and/or prolonged physical exercise caused only a modest increase of FDP/fdp. Physical training seems only minimally to affect this response. It is speculated that in the trained subject, daily exercise-induced fibrinolytic activation might be more relevant to the risk of thrombotic or atherosclerotic disease than a higher absolute resting or post-exercise fibrinolytic level compared with the untrained.

Complement activation after prolonged exercise.

The effects of a 2.5-h running test on the concentrations of the complement cleavage products C3a, C4a and C5a in plasma were examined in eight healthy young males. In all participants the C3a and C4a values were raised during (1 h after the start) and immediately after exercise. C4a was also raised 1 and 3 h after the race. These results provide evidence for complement activation after prolonged exercise.

Assessment of circulating immune complexes by a solid-phase C1q-binding assay during the first hours and days after prolonged exercise.

A sensitive and specific solid-phase C1q-binding assay using porcine C1q and microtitre plates as a solid phase, served to assess the immune complexes in serum during the first hours and days after a 3-h running test. Fourteen moderately trained male subjects participated in the race and covered 36.3 +/- 3.7 (mean +/- SD) km in 3 h. Blood samples were drawn 1 day before, immediately before, immediately after, 1 and 3 h after and 1, 2 and 4 days after the race. When corrected for the changes of serum total protein, the apparent immune complex concentrations were 1 and 3 h after the race significantly higher (p less than 0.001 and p less than 0.05 respectively) than the pre-exercise values. Two days after the race the values were significantly lower (p less than 0.05) than before the race. These findings provide evidence for the formation of immune complexes after severe physical exercise.

Rapid DNA typing of HLA-B27 allele by real-time PCR using lightcycler technology.

For clinical diagnostic routine we developed a fast DNA typing of HLA-B27 by PCR and real-time detection using LightCycler technology. The method combines the sensitivity and specificity of PCR with the swiftness of the LightCycler system. The amplification step was performed with a primer set coding for a region in the third exon common to B*2701 to B*2705. The PCR cycles were monitored continuously using the SYBR Green I dye. Beta-globin was used as an internal control. An analysis of 32 samples with one PCR run was completed within 40 minutes. After amplification a melting curve analysis permitted the accurate identification of the PCR amplicons. The mean melting temperatures (Tm) were 90.5 degrees C and 87.3 degrees C, which are characteristic for HLA-B27 and beta-globin, respectively. A comparison of 300 samples which were typed for HLA-B27 with a conventional sequence-specific polymerase chain reaction (SSP-PCR) and with the new method demonstrated a perfect correlation (specificity 100%). In summary, the method described is fast, reliable, cost-effective and well adapted for routine laboratory testing.

Plasma elastase-alpha 1-antitrypsin, neopterin, tumor necrosis factor, and soluble interleukin-2 receptor after prolonged exercise.

The effects of a 2.5-h running test on the plasma concentrations of elastase-alpha 1-antitrypsin, neopterin, tumor necrosis factor, and soluble interleukin-2 receptor were evaluated in eight healthy young male subjects. Neopterin was measured by radioimmunoassay, elastase-alpha 1-antitrypsin, tumor necrosis factor, and soluble interleukin-2 receptor by enzyme immunoassay. The post-exercise values were corrected for plasma volume changes which were calculated from hematocrit and hemoglobin values. Compared with the concentrations before exercise, elastase-alpha 1-antitrypsin values were significantly increased during the run (1 h after the start) (P less than 0.01) as well as during the first few hours after the end of the running test (P less than 0.01). A significant increase of tumor necrosis factor and neopterin was observed 1 h after the end and 1,3, and 24 h after the end of the running test, respectively, (P less than 0.01), and soluble interleukin-2 receptor concentrations were significantly elevated 1 and 2 days after exercise (P less than 0.01). The increase of elastase-alpha 1-antitrypsin, neopterin, tumor necrosis factor, and soluble interleukin-2 receptor supports the concept of a functional involvement of polymorphonuclear neutrophils and an activation of macrophages and T-lymphocytes after prolonged exercise.

Effect of a short maximal physical exercise on coagulation, fibrinolysis, and complement system.

In 11 healthy young subjects, the plasma concentrations of the thrombin-antithrombin III complex, fibrinopeptide A, tissue-plasminogen activator, complement fragments C3a and C4a, and histamine were measured before and after a graded maximal bicycle exercise test. The analyses were carried out 30 min before and immediately before exercise, immediately after exercise, and 30 and 60 min later. All post-exercise values were corrected for plasma volume changes, which were calculated from hematocrit and hemoglobin values. Immediately post-exercise, thrombin-antithrombin III, tissue-plasminogen activator, complement fragments C3a and C4a, and histamine were all significantly elevated (p less than 0.01), compared with the pre-exercise values; 30 and 60 min later the values normalized and significant differences from the pre-exercise values could no longer be measured. Fibrinopeptide A did not change significantly after exercise. The present results provide evidence for a simultaneous activation of coagulation, fibrinolysis, and complement system as well as for a release of histamine after a short maximal exercise.

Competitive enzyme immunoassay for apolipoprotein A-II.

A competitive enzyme immunoassay for apolipoprotein A-II was developed. Microtitre plates were used as a solid phase and coated with anti-apolipoprotein A-II antibodies. Purified apolipoprotein A-II, labelled with horseradish peroxidase was used as competing ligand. The assay was examined with respect to the optimal amounts of specific anti-apolipoprotein A-II antibodies and apolipoprotein A-II-enzyme conjugate. The displacement curves showed a good parallelism between serum and purified apolipoprotein A-II. Delipidation of serum did not affect the content of apolipoprotein A-II. Cross-reactivity with apolipoprotein A-I was minimal. The intra- and inter-assay coefficients of variation in the medium range were 9.0 and 11.8% respectively. The assay might be well-suited for clinical routine.

Modifications of serum glycoproteins the days following a prolonged physical exercise and the influence of physical training.

Eight male subjects (mean age 24.1 +/- 2.6 years) performed at intervals of 2 weeks successively a 3 h and two 2 h runs of different running speed. The days following the running there were moderate elevations of C-reactive protein, haptoglobin, alpha-1-acid glycoprotein, coeruloplasmin, transferrin, alpha-1-antitrypsin and plasminogen. There were small or no changes of albumin, alpha-2-macroglobulin and hemopexin. The elevations of the "acute phase reactants" were examined in three male subjects following a 2 h run before and after an endurance training period of 9 weeks. This demonstrated a decreased acute phase response after training as illustrated by the changes of C-reactive protein, haptoglobin and alpha-1-acid glycoprotein in spite of higher posttraining running speeds. Well-trained athletes have elevated levels of the serum protease inhibitors alpha-1-antitrypsin, alpha-2-macroglobulin and C1-inhibitor. These antiproteolytic glycoproteins might limit exercise-induced inflammatory reactions.

The delayed effects of prolonged physical exercise and physical training on cholesterol level.

The delayed effects of prolonged physical exercise on total and high-density lipoprotein (HDL) cholesterol were studied before and after a training period of 8 weeks. In 15 subjects, 1 and 2 days after a 3 h running test, total cholesterol was significantly lower than 1 day before and 8 days after the end of the exercise. One day post-exercise, HDL-cholesterol was significantly higher compared with 1 day before and 8 days afterwards. In ten subjects participating in the training program, decreased total cholesterol values were again found on the first day after the post-training exercise test. Total and HDL cholesterol levels were not significantly affected by 8 weeks of training.

Lymphocyte subsets during the first hours and days after a 2.5 h running test.

After a 2.5 h running test on eight healthy young males the lymphocyte subsets in peripheral blood were analyzed by flow cytometry. The analysis was carried out immediately post-exercise, after a 1 h and 3 h interval as well as a one day, a two day and five day interval. Lymphocyte subpopulations were also measured in five control subjects who did not exercise. Compared with the pre-exercise level the total lymphocyte and T-cell count was decreased 1 h and 3 h after the run and one and two days after exercise (p less than 0.01). The absolute number of monocytes and B-lymphocytes fell significantly (p less than 0.01) on the first and second day after exercise. No consistent change of helper-/suppressor-inducer T-lymphocytes (CD4+) and cytotoxic-suppressor T-cells (CD8+) cells was observed immediately after exercise. One and 3 h later, however, the absolute number of both cell types dropped below pre-exercise values (p less than 0.01). The decrease of the CD8+ cells was more pronounced, causing an increase of the CD4+ to CD8+ ratio (p less than 0.01). Among the CD4+ cells only the Leu 8+ subpopulation showed a relative increase 1 h after the exercise (p less than 0.01) while the Leu 8- population remained unaffected. 1 h and 3 h after the end of the run there was a significant fall of the absolute number and percentage of natural killer cells (Leu 7+) and cytotoxic T-cells (CD8+ Leu 7+) (p less than 0.01). These cells remained low during the first and second day after the exercise (p less than 0.01). In the control subjects the changes of cell counts for all cells examined were much smaller than in the runners. The results indicate that strenuous and prolonged exercise causes a delayed redistribution of lymphocyte subpopulations with a reduction of lymphocytes and particularly of natural killer and cytotoxic cells beginning 1 h after the exercise followed by a reduction of monocytes 24 h later. The percentage increase of CD4+ cells 1 h after the exercise could be explained by an increase of the CD4+ Leu 8+ subpopulation.

C-reactive protein serum concentrations in well-trained athletes.

Previously it had been shown that there is a sharp increase in C-reactive protein (CRP) serum concentration during the first few days after severe exercise. To evaluate the effect of physical training on the CRP levels, a sensitive enzyme immunoassay was developed and used to assess the basal CRP concentrations in 356 male and 103 female athletes, who trained at least 4 times per week in various disciplines, in 45 male and 40 female untrained controls, and in 35 elderly coronary patients. In male athletes the lowest CRP values were found in swimmers (median: 102 ng/ml, n = 59), which were significantly lower (P less than 0.001) than those of male controls (median: 502 ng/ml, n = 45). The CRP concentrations in rowers (median: 260 ng/ml, n = 108) were also significantly lower (P less than 0.01) than in the control group. CRP levels in middle- and long-distance runners (median: 315 ng/ml, n = 53), racing cyclists (median: 620 ng/ml, n = 111), and soccer players (median: 660 ng/ml, n = 25) did not differ significantly from those of the controls. In female athletes the lowest CRP concentrations were also found in swimmers (median: 110 ng/ml, n = 49), which were significantly lower (P less than 0.001) than those of the female controls (median: 396 ng/ml, n = 40). Female middle- and long-distance runners (n = 36) and female rowers (n = 18) had median CRP values of 250 and 285 ng/ml, respectively. This results suggest that training induces a suppressive effect upon CRP, responsible for the low serum levels in athletes (particularly swimmers) when compared to controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of a short maximal physical exercise on the eosinophil cationic protein.

Eosinophil cationic protein (ECP) has been shown to be a marker of eosinophil granulocyte activation. In 10 healthy young subjects the plasma concentrations of ECP were measured before and after a graded maximal bicycle exercise test. The analyses were carried out 30 min before and immediately before exercise, immediately after exercise and 20 and 45 min later. The post-exercise values were corrected for plasma volume changes which were calculated from hematocrit and hemoglobin values. Immediately post-exercise the ECP increased significantly (p < 0.01) from 1.25 +/- 0.28 at rest to 2.40 +/- 0.59 micrograms/l. Twenty and 45 min later the values normalized and significant differences from the pre-exercise values could no longer be measured. The results provide strong evidence for an activation of eosinophil granulocytes after a short maximal exercise.

Serum ferritin, transferrin, haptoglobin, and iron in middle- and long-distance runners, elite rowers, and professional racing cyclists.

Serum ferritin, transferrin, haptoglobin, and iron were measured in well-trained middle- and long-distance runners, elite rowers of the West German national team, and professional racing cyclists during the summer training and the winter rest period. None of the male athletes examined, with the exception of the racing cyclists during the summer period, received oral or parenteral iron. The runners were found to have significantly lower ferritin (P less than 0.00001), iron (P less than 0.001), and haptoglobin values (P less than 0.01) than the controls. Their transferrin levels were elevated, however not significantly. Rowers showed significantly higher ferritin levels (P less than 0.01) than the controls. The reduced haptoglobin concentrations in runners are presumed to be caused by a running-induced hemolysis. It is speculated that a recurring hemoglobinuria produced diminished iron reserves in middle- and long-distance runners.

Blood glutathione status following distance running.

In 12 moderately trained subjects reduced glutathione (GSH) and oxidized glutathione (GSSG) as well as thiobarbituric acid reactive substances (TBARS) were measured in the blood before and during the first two hours and first two days after a 2.5-h run. The participants covered between 19 and 26 km (20.8 +/- 2.5 km, mean +/- SD). The running speed was between 53 and 82% of the speed at which blood lactate concentration reached 4 mmol/L lactate (67.9 +/- 8.2%, mean +/- SD) assessed during a previously performed treadmill test. Blood samples were collected 1 h before, immediately before, immediately after, 1 and 2 h after, as well as 1 and 2 days after the run. Immediately after exercise GSH was significantly decreased (p < 0.01) and GSSG significantly increased (p < 0.01). In all subjects the ratio of GSH to GSSG showed a marked decline to 18 +/- 4% (mean +/- SD) of the pre-exercise values (p < 0.01). One hour later the mean GSH and GSSG values returned to baseline. However, there were considerable inter-individual differences. In some subjects the GSH/ GSSG ratio overshot the pre-exercise levels, in others the ratio remained low even two hours after exercise. Compared with the pre-exercise values TBARS concentrations did not change significantly at any time point after exercise. The findings suggest that after prolonged exercise in moderately trained subjects a critical shift in the blood glutathione redox status may be reached. The changes observed were generally short-lived, the duration of which may have depended on the relative importance of reactive oxygen species generation by the capillary endothelial cells and neutrophil and eosinophil granulocytes after the end of exercise.

Reduction of the plasma concentration of C-reactive protein following nine months of endurance training.

An intense physical exercise induces an inflammatory reaction as demonstrated by the delayed increase in blood of acute phase proteins and among them of C-reactive protein (CRP). There is also evidence for a diminished acute phase reaction due to regular exercise suggesting a suppression of the inflammatory response through training. With this background CRP was measured by a sensitive enzyme immunoassay under resting conditions before and after 9 months of training in 14 subjects preparing for a marathon with the aim of studying the effect of training on the base-line CRP concentration. The mean distance run per week increased significantly from 31 +/- 9 km at the beginning to 53 +/- 15 km after 8 months of training (p < 0.01). The aerobic capacity rose significantly after training as demonstrated by the increase of running velocity during a maximal treadmill test from 3.82 +/- 0.29 m/s pre-training to 4.17 +/- 0.17 m/s post-training at a blood lactate concentration of 4 mmol/L (p < 0.01). In 10 of 12 runners base-line CRP was diminished after training in spite of a continuous increase of training intensity. The CRP median fell from 1.19 mg/L before to 0.82 mg/L after training (p < 0.05). Since intense physical exercise is known to be associated with an inflammatory reaction of muscles and tendons, the CRP decrease was unexpected. In 2 subjects the CRP concentration rose markedly because of a borrelia infection and a knee injury, respectively. These values were caused by a pathological condition and were not considered for the statistical evaluation. In 10 non-training control subjects the CRP median did not change significantly during the same 9 months period. The decrease of the CRP base-line concentration after training suggests that intensive regular exercise has a systemic anti-inflammatory effect. This is of particular interest with regard to several recent reports confering on the concentration of CRP in plasma a predictive value for the risk of cardiac infarction, venous thrombosis or stroke.