Relation between Exercise Performance and Blood Storage Condition and Storage Time in Autologous Blood Doping

Professional athletes are expected to continuously improve their performance, and some might also use illegal methods—e.g., autologous blood doping (ABD)—to achieve improvements. This article applies a systematic literature review to investigate differences in the ABD methods and the related performance and blood parameters owing to different storage conditions—cryopreservation (CP) and cold storage (CS)—and different storage durations. The literature research resulted in 34 original articles. The majority of currently published studies employed CS during ABD. This contrasts to the applied storage technique in professional sports, which was mainly reported to be CP. The second outcome of the literature research revealed large differences in the storage durations applied, which were in the range of one day to 17 weeks between blood sampling and re-infusion, which might affect recovery of the red blood cell mass and thus performance outcome related to ABD. Data revealed that performance parameters were positively affected by ABD when a minimal storage duration of four weeks was adhered. This article identified a need for further research that reflect common ABD practice and its real effects on performance parameters, but also on related blood parameters in order to develop valid and reliable ABD detection methods.

Monitoring of RBC rheology after cryopreservation to detect autologous blood doping in vivo? A pilot study

BACKGROUND: Autologous blood doping (ABD) is applied to improve performance capacity. ABD includes blood donation, red blood cell (RBC) storage at –80°C and re-infusion prior to or during competition. ABD is not directly detectable with current detection techniques. OBJECTIVE: Since cryopreservation is known to affect RBC physiology in vitro, the aim of the study was to examine whether these alterations are detectable in vivo. METHODS: Blood from six healthy male donors was transferred into conventional blood bags, cryopreserved, stored for 18 weeks at –80°C and re-infused with a RBC volume corresponding to ∼4% of total blood volume into respective donor. RBC physiology parameters were measured before blood donation/re-infusion, and 0/1/2/6/24/48/72 h and 1 w post re-infusion. RESULTS: RBC parameters and age markers were unaffected during intervention. RBC deformability increased from pre-blood-sampling to pre-re-infusion while deformability and viscosity values remained unaltered post re-infusion. RBC nitric oxide associated analytes, metabolic parameters and electrolyte concentrations remained unaffected. CONCLUSIONS: The data of this pilot study indicate that the increase in RBC deformability might be related to neoformation of RBC after blood donation. The lack of changes in tested parameters might be related to the low re-infused RBC volume which might explain differences to in vitro results.

DETECTION OF AUTOLOGOUS BLOOD DOPING

ObjectivesAutologous blood transfusion (ABT) enhances athlete’s performance, is banned as doping by the World Anti-Doping Agency (WADA). Currently, there is no implemented detection method for ABT. Transfusion of one’s own, long-term cryopreserved red blood cells (cryo-RBC) immediately increases circulating RBC count, hemoglobin mass, blood volume and oxygen carrying capacity, resulting in enhanced physical performance. Functional viablity of cryo-RBC are maintained for decades, but storage lesions lead to removal of damaged RBC from circulation days after transfusion, with remaining circulating cryo-RBC displaying normal half-life.MethodsThe cytosolic RBC peptidome from 22 human subjects (12 men and 10 women) was analyzed by UHPLC-MS/MS before and after ABT with cryo-RBC. As a control group and for investigation of confounders, 14 elite athletes and 5 recreational subjects were sampled multiple times, also at high altitude.ResultsHere we report alteration in the cytosolic peptidome of circulating RBC weeks after ABT, discriminating doped from non-doped human subjects. A valid discriminating multivariate model (OPLS-DA) based on <200 peptides was accomplished (R2/Q2 = 0.88/0.59, P CV-ANOVA < 0.0001, ROC AUC = 0.97). Models did not show bias for sex, high altitude or elite endurance training and racing.ConclusionIdentified peptides with low intra- and inter-individual variation, and high multivariate model weight and probability scores, create a direct method for the detection of autologous blood doping.

Automation of RNA-based biomarker extraction from dried blood spots for the detection of blood doping

Aim: Transcriptomic biomarkers originating from reticulocytes measured in dried blood spots (DBSs) may be reliable indicators of blood doping. Methods/results: Here, we examined changes in the expression levels of the erythropoiesis-related ALAS2, CA1 and SLC4A1 genes in DBS samples from elite athletes and volunteers of clinical study with recombinant erythropoietin dose. Conclusion: By comparing the mean intraday coefficients of variation for ALAS2L, ALASLC, CA1 and SLC4A1 between manual and automated RNA extractions, an average improvement was observed, whereas the assessment of interday variability provided comparable results for both manual and automated approaches. Our results confirmed that RNA biomarkers on DBS support are efficient to detect blood doping.

Prevalence estimate of blood doping in elite track and field at the introduction of the Athlete Biological Passport

In elite sport, the Athlete Biological Passport (ABP) was invented to tackle cheaters by monitoring closely changes in biological parameters, flagging atypical variations. The haematological module of the ABP was indeed adopted in 2011 by the International Association of Athletics Federations (IAAF). This study estimates the prevalence of blood doping based on haematological parameters in a large cohort of track & field athletes measured at two international major events (2011 & 2013 IAAF World Championships) with a hypothesized decrease in prevalence due to the ABP introduction.A total of 3683 blood samples were collected and analysed from all participating athletes originating from 209 countries. The estimate of doping prevalence was obtained by using a Bayesian network with seven variables, as well as “doping” as a variable mimicking doping with low-doses of recombinant human erythropoietin (rhEPO), to generate reference cumulative distribution functions (CDFs) for the Abnormal Blood Profile Score (ABPS) from the ABP.Our results from robust haematological parameters indicate an estimation of an overall blood doping prevalence of 18% in average in endurance athletes (95% Confidence Interval (C.I.) 14-22%). A higher prevalence was observed in female athletes (22%, C.I. 16-28%) than in male athletes (15%, C.I. 9-20%). In conclusion, this study presents the first comparison of blood doping prevalence in elite athletes based on biological measurements from major international events that may help scientists and experts to use the ABP in a more efficient and deterrent way.What are the new findings ?This study presents the first comparison of blood doping prevalence in elite track & field athletes based on biological measurements from major international eventsOur results from robust haematological parameters indicate an estimation of an overall blood doping prevalence of 18% in average in endurance athletes.The confidence intervals for blood doping prevalence range from 9-28% with wide discrepancies between certain countries.How might it impact on clinical practice in the near futureThe further development of the Athlete Biological Passport with a careful monitoring of biological parameters still represents the most consistent approach to thwart athletes using undetectable prohibited substances or methods.This study describes a method to define blood doping prevalence with the analysis of robust haematological parametersEstimates of doping prevalence in subpopulations of athletes may represent a valuable tool for the antidoping authorities to perform a risk assessment in their sport.