Plasma centrifugation does not influence thrombin-antithrombin and plasmin-antiplasmin levels but determines platelet microparticles count

Introduction

Centrifugation is an essential step for plasma preparation to remove residual elements in plasma, especially platelets and platelet-derived microparticles (PMPs). Our working hypothesis was that centrifugation as a preanalytical step may influence some coagulation parameters.

Materials and methods

Healthy young men were recruited (N = 17). For centrifugation, two protocols were applied: (A) the first centrifugation at 2500 x g for 15 min and (B) at 2500 x g for 20 min at room temperature with a light brake. In protocol (A), the second centrifugation was carried out at 2500 x g for 15 min, whereas in protocol (B), the second centrifugation involved a 10 min spin at 13,000 x g. Thrombin-antithrombin (TAT) and plasmin-antiplasmin (PAP) complexes concentrations were determined by enzyme-linked immunosorbent assays. PMPs were stained with CD41 antibody and annexin V, and analyzed by flow cytometry method. Procoagulant activity was assayed by the Calibrated Automated Thrombogram method as a slope of thrombin formation (CAT velocity).

Results

Median TAT and PAP concentrations did not differ between the centrifugation protocols. The high speed centrifugation reduced the median (IQR) PMP count in plasma from 1291 (841-1975) to 573 (391-1010) PMP/µL (P = 0.001), and CAT velocity from 2.01 (1.31-2.88) to 0.97 (0.82-1.73) nM/min (P = 0.049). Spearman’s rank correlation analysis showed correlation between TAT and PMPs in the protocol A plasma which was (rho = 0.52, P < 0.050) and between PMPs and CAT for protocol A (rho = 0.74, P < 0.050) and protocol B (rho = 0.78, P < 0.050).

Conclusion

Centrifugation protocols do not influence the markers of plasminogen (PAP) and thrombin (TAT) generation but they do affect the PMP count and procoagulant activity.Key words: cell-derived microparticles, blood coagulation tests, centrifugation, preanalytical phase     

Physiological Correlations With Short,Medium, and Long Cycling Time-Trial Performance

Purpose: Several studies have demonstrated that physiological variables predict cycling endurance performance. However, it is still unclear whether the predictors will change over different performance durations. The aim of this study was to assess the correlations between physiological variables and cycling time trials with different durations. Methods: Twenty trained male cyclists (maximal oxygen uptake [VO₂max] = 60.5 ± 5.6 mL/kg/min) performed 4 separate experimental trials during a 2-week period. Cyclists initially completed an incremental exercise test until volitional exhaustion followed by 3 maximal cycling time trials on separate days. Each time trial consisted of 3 different durations: 5 min, 20 min, and 60 min performed in a randomized order. Results: The main results showed that the physiological measures strongly correlated with long cycling performances rather than short and medium time trials. The time-trial mean power output was moderately high to highly correlated with peak power output and VO_2max (r = .61–.87, r = .72–.89, respectively), and was moderately to highly correlated with the lactate threshold Dmax method and second ventilatory threshold (r = .52–.75, r = .55–.82, respectively). Conclusions: Therefore, trained cyclists should develop maximal aerobic power irrespective of the duration of time trial, as well as enhancements in metabolic thresholds for long-duration time trials.