Platelet aggregation and strenuous exercise

1. Platelet aggregation in the Chandler's tube has been found to be increased in a group of normal male and female volunteers who undertook strenuous physical exercise. This coincided with acceleration of the ;intrinsic' blood clotting system and a rise in fibrinogen. The rise in fibrinogen occurred despite increased fibrinolysis.2. The study confirms the sensitivity of the platelet aggregation system to changes in the ;intrinsic' clotting mechanism. Acceleration of this system in this study resulted from a physiological cause and produced accelerated aggregation in the coagulation-affected phase.     

Platelet adhesion and activation on a shielded plasma gradient prepared on polyethylene.

Contact of blood with foreign materials evokes thrombogenic effects to an extent determined partly by the wettability of the biomaterials surface. Tools to study blood response towards a variation in materials wettability with minimal variation in chemistry are "gradient surfaces". However, most gradients have been prepared by diffusion or density immersion techniques, which results in a limited gradient range. Through glow discharge with partial shielding, gradients on polymers were prepared over a length of 5 cm, which facilitated studies to platelet adhesion on separate gradient sections. On polyethylene, advancing water contact angles varied from 90 degrees to 40 degrees, with a hysteresis of 30 degrees. ESCA indicated an increasing incorporation of oxygen towards the hydrophilic end. To examine the role of materials wettability on the activation of adhering platelets, sections of shielded plasma gradients were incubated in anticoagulated whole human blood. Fewer platelets adhered to the hydrophobic end, but those platelets were more activated than those on the hydrophilic end, as judged from their morphology and exposure of GpIIb-IIIa complex. However, partly related to the increased binding of platelets, the clotting activation after platelet deposition was highest on the hydrophilic end. Concluding, this new technique results in a large gradient range, which facilitates studies of formed blood elements in relation to the wettability. Platelets are more activated on hydrophobic polyethylene, while on moderate hydrophilic polyethylene more platelet adhesion and activation of the clotting system occurs.     

Platelet adhesion on titanium oxide gels: effect of surface oxidation

The correlations between titanium oxide layers on oxidized titanium (Ti) substrates and platelet adhesion were examined. Ti substrates were prepared by three different oxidation methods: the first one was treated with hydrogen peroxide (H2O2) solution, the second one was heated in air at moderate temperatures, and the third one was processed with both H2O2 and heating. The titanium oxide layers formed on the Ti substrates were characterized by wettability, chemical composition, thickness, and crystal phase. The platelet adhesion on these oxide layers was examined and correlated to the characterizations of the surface layers. The number of adhesive platelets seemingly correlated with the contact angle towards distilled water, because the number increased close to 70-80 degrees of the contact angle. The effect of surface oxidation on platelet adhesion was examined in detail and it was found that the composition and thickness of the oxide layer influenced platelet adhesion rather than wettability. Thick titanium oxide layers formed on Ti substrates by heating displayed less platelet adhesion than thin oxide layers on untreated Ti substrates. The largest number of adhesive platelets was found on H2O2-oxidized substrates; the substrates found on amorphous titanium oxide contained the Ti-peroxide radical species. The number of platelets, on the other hand, could hardly be observed on Ti substrates treated with H2O2 and subsequently heated above 300 degrees C. The titanium oxide layer on the substrate was thick and we found it to consist of only a few radical species. That is, the effect of heat treatment accelerates the growth of the oxide layer, and decomposes or decreases the number of radical species. Ti substrates with H2O2 and heat treatment above 300 degrees C held the least number of platelets, and were concluded to be the most inhibitory for platelet adhesion.     

The effect of various levels of strenuous to exhaustive exercise on reaction time

Reaction times and heart rates of 10 subjects walking and running on a motor-driven treadmill were recorded. The exercise consisted of four 3-min stages: Stage 1, 2.5 mph at 12% grade; Stage 2, 3.4 mph at 14% grade; Stage 3, 4.2 mph at 16% grade; Stage 4, 5.0 mph at 18% grade. Subjects were given three separate tests at 1-week intervals. There was an increase in reaction time with increased exercise, but this effect diminished significantly over days. Also, when heart rate was 80% of maximum during the post-test, reaction time was not significantly different from the pre-test. Optimum reaction time was obtained at approximately 40% of maximum heart rate.     

Carbon coated polyethylene: effect of surface energetics and topography on human platelet adhesion

The influence of surface energy and structural properties of carbon coated polyethylene (PE) on the human platelet adhesion was studied. Three types of amorphous carbon coating were obtained by plasma pulse discharge, with the number of pulses grading as 10, 50, 100. Human serum albumin adsorption experiments have been carried out with all samples in vitro. Platelet adhesion analysis by SEM included determination of total quantity of adherent platelets, and respective quantities of platelets at different stages of activation (single, spread, aggregates). Surface topographies ranged from bare PE and such (10 pulses), to globular 0.5 microm in size (50 pulses), and complex fibrillar 3-4 microm structures (100 pulses). Surface free energy varies from 31.7 +/- 0.6 to 40.4 +/- 0.6 mN/m for uncoated PE and 10 pulse coatings, respectively, as determined by contact angle techniques. All studied coatings demonstrate weaker platelet activation properties in comparison with untreated PE. Among all studied coatings, the 50 pulse coated surface seems to be the least suitable for contact with platelets, mainly due to its structural rather than to its energy properties. These data are related to a sharp decrease in the adsorbed protein level for the samples with 50 pulse coatings. The applied analysis of platelet activation enables more accurate characterization of platelet-biomaterial interaction.     

Effects of exhaustive stretch-shortening cycle exercise on muscle blood flow during exercise

Aim: The influence of exhaustive stretch‐shortening cycle exercise (SSC) on skeletal muscle blood flow (BF) during exercise is currently unknown. Methods: Quadriceps femoris (QF) BF was measured in eight healthy men using positron emission tomography before and 3 days after exhaustive SSC exercise. The SSC protocol consisted of maximal and submaximal drop jumps with one leg. Needle biopsies of the vastus lateralis muscles were taken immediately and 2 days after SSC for muscle endothelial nitric oxide synthase (eNOS) and interleukin‐1‐beta (IL‐1β) mRNA level determinations. Results: All subjects reported subjective muscle soreness after SSC (P < 0.001), which was well in line with a decrease in maximal isometric contraction force (MVC) and increase in serum creatine kinase activity (CK) (P = 0.018). After SSC muscle BF was 25% higher in entire QF (P = 0.043) and in its deep and superficial muscle regions, whereas oxygen uptake remained unchanged (P = 0.893). Muscle biopsies revealed increased IL‐1β (30 min: 152 ± 75%, P = 0.012 and 2 days: 108 ± 203%, P = 0.036) but decreased or unchanged eNOS (30 min; ?21 ± 57%, P = 0.050 and 2 days: +101 ± 204%, P = 0.779) mRNA levels after SSC. Conclusion: It was concluded that fatiguing SSC exercise induces increased muscle BF during exercise, which is likely to be associated with pro‐inflammatory processes in the exercised muscle.     

Plasma glucagon and catecholamines during exhaustive short-term exercise

Summary Plasma glucagon and catecholamine levels were measured in male athletes before and after exhaustive 15 min continuous running and strenuous intermittent short-term exercise (3×300 m). Blood lactate levels were higher after the intermittent exercise (mean 16.7 mmol×l^–1) than after the continuous running (mean 7.1 mmol×l^–1). Plasma glucagon concentration increased during continuous running and intermittent exercise by 41% and 55%, respectively, and the increases in plasma noradrenaline concentration were 7.7- and 9.1-fold compared with the respective pre-exercise values. Immediately after the exercises plasma cyclic AMP, blood glucose and alanine levels were elevated significantly.The data suggest that the sympathoadrenal system is of major importance for liver glucose production during high-intensity exercises. Catecholamines directly stimulate liver glucose production and may indirectly stimulate it by enhancing the secretion of glucagon.     

The effect of exhaustive exercise on the antioxidant enzyme system in skeletal muscle from calcium-deficient rats

 The aim of the current study was to elucidate the synergism of dietary calcium restriction and exhaustive exercise in the antioxidant enzyme system of rat soleus muscle, and to investigate the involvement of neutrophils in exercise-induced muscle damage. Forty-eight male Wistar rats were assigned to the following groups: control (C) or calcium-restricted [1 month (1 M) or 3 months (3 M)]. Each group was subdivided into acutely exercised or non-exercised groups. Soleus muscle from each rat was analysed to determine the levels of antioxidant enzymes [Mn-superoxide dismutase (SOD), Cu,Zn-SOD, glutathione peroxidase (GPX), and catalase (CAT)]. Dietary calcium restriction resulted in calcium deficiency and upregulated the antioxidant enzymes examined except GPX. Conversely, exhaustive exercise significantly decreased GPX and CAT, but not SODs activities in the calcium-restricted (1 M and/or 3 M) rats. Contents of immunoreactive Mn-SOD and Cu,Zn-SOD were only increased in the 3 M rats. During calcium restriction, the mRNA expression of both forms of SOD showed initial upregulation, followed by downregulation. Exhaustive exercise significantly increased the mRNA expressions only in the 3 M rats. Moreover, exhaustive exercise markedly increased myeloperoxidase activity in soleus muscles from the 1 M and 3 M rats compared with the C rats, and significantly enhanced the ability of neutrophils to generate superoxide in the 3 M rats. The results demonstrate that dietary calcium restriction upregulates certain antioxidant enzyme activities in rat soleus muscle, indicating an enhanced resistance to potential increases in intracellular reactive oxygen species. The results also suggest that exhaustive exercise may cause oxidative damage in soleus muscle of calcium-deficient rats through the activation of neutrophils. Received: 4 August 1997 / Received after revision: 29 September 1997 / Accepted: 26 November 1997     

Platelet adhesion onto polyetherurethane urea derivatives: effect of cytoskeleton proteins of the platelet

Adhesion and activation of platelets upon adhesion onto synthetic polymers were investigated with reference to participation of the cytoskeleton proteins. Platelets were treated with cytoskeleton breakers, and then the adhesion of platelets onto polyetherurethane urea derivatives and serotonin release from adhered platelets were investigated. In the adhesion onto glass, platelets were strongly stimulated and accompanied rearrangement of the cytoskeleton system, and also serotonin release involved the action of the cytoskeleton system. On the other hand, platelets were not strongly stimulated upon adhering onto polyetherurethane urea derivatives. The platelet adhesion onto cationic polymers exceptionally accompanied the rearrangement of the cytoskeleton system. The participation of the cytoskeleton in platelet adhesion onto polyetherurethane urea derivatives was influenced by the presence of plasma proteins. It was found that protein layers deposited on the material surface play an important role in platelet adhesion.     

The VO2 response to exhaustive square wave exercise: influence of exercise intensity and mode

We investigated the oxygen uptake (O₂) response to exhaustive square wave exercise of approximately 2, 5 and 8 min duration in cycling and running. Nine males completed a ramp test and three square wave tests on a motorised treadmill and the same four tests on a cycle ergometer, throughout which gas exchange was assessed (Douglas bag method). The peak O₂ from the ramp test was higher for running than for cycling [mean (SD): 58.4 (2.8) vs. 55.9 (3.7) ml.kg^–1.min^–1; P=0.04]. However O_2max (defined as the highest O₂ achieved in any of the four tests) did not differ between running and cycling [60.0 (2.9) vs. 58.5 (3.3) ml.kg^–1.min^–1; P=0.15]. The peak O₂ was similar (P>0.1) for the 5 and 8 min square wave tests [98.5 (1.8) and 99.2 (2.3) %O_2max for running; 97.0 (4.2) and 97.5 (2.0) %O_2max for cycling] but lower (P<0.001) for the 2-min test [91.8 (2.5) and 89.9 (5.5) %O_2max for running and cycling respectively]. O₂ increased over the final two 30-s collection periods of the 2-min test for cycling [O₂=0.18 (0.15) l.min^–1; P<0.01] but not running [O₂=0.00 (0.09) l.min^–1; P=0.98]. We conclude that in the aerobically fit the peak O₂ for square wave running or cycling at an intensity severe enough to result in exhaustion in approximately 2 min is below O_2max. In running, O₂ plateaus at this sub-maximal rate.