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We investigated the effects of recombinant human erythropoietin (rHuEPO) administration on exercise endurance, maximal aerobic performance and total hemoglobin mass (tHb). We hypothesized that frequent, small intravenous injections of epoetin β would increase time trial performance, peak oxygen uptake (V̇O2peak) and tHb in both males and females.We included 48 healthy, recreational to trained males (n = 24, mean ± standard deviation V̇O2peak of 55 ± 5 ml O2 × kg-1 × min-1) and females (n = 24; V̇O2peak of 46 ± 4 ml O2 × kg-1 × min-1) in a counter-balanced, double-blind, randomized, placebo-controlled study design stratified by sex. Time trial performance, V̇O2peak, and tHb were determined before and after intravenous injections of either rHuEPO (9 IU × kg bw-1 epoetin β) or saline (0.9% NaCl) three times weekly for 4 weeks.A time × treatment effect (P0.05) existed for time trial performance. Within the rHuEPO group, mean power output (MPO) increased by 4.1 ± 4.2% (P0.001). Likewise, a time × treatment effect (P0.001) existed for V̇O2peak, where the rHuEPO group improved V̇O2peak and peak aerobic power by 4.2 ± 6.1% (P0.001) and 2.9 ± 4.0% (P0.01), respectively. A time × treatment effect (P0.001) existed for tHb, where the rHuEPO group increased tHb by 6.7 ± 3.4% (P0.001). A main effect of 'sex' alone was also evident (P0.001) but no sex-specific interactions were found. No changes were observed in the placebo group for MPO, V̇O2peak, peak aerobic power or tHb.Micro-doses with intravenous rHuEPO provide a sufficient erythropoietic stimuli to augment tHb and enhance aerobic-dominated performance in both trained males and females.

Purpose: We investigated whether hepcidin and erythroferrone (ERFE) could complement the Athlete Biological Passport (ABP) in indirectly detecting a 130 mL packed red blood cells (RBCs) autologous blood transfusion. Endurance performance was evaluated.Methods: Forty-eight healthy men (n = 24) and women (n = 24) participated. Baseline samples were collected weekly followed by randomization to a blood transfusion (BT, n = 24) or control group (CON, n = 24). Only the BT group donated 450 mL whole blood from which 130 mL RBCs was reinfused four weeks later. Blood samples were collected 3, 7, 14, 21 and 28 days after donation, and 3, 6, and 24 hours and 2, 3, and 6 days following reinfusion. In the CON group samples were collected with the same frequency. Endurance performance was evaluated by a 650-kCal time trial (n = 13) before and one and six days after reinfusion.Results: A time×treatment effect existed (P < 0.05) for hepcidin and ERFE. Hepcidin was increased (P < 0.01) ~110 and 89% six and 24 hours after reinfusion. Using an individual approach (99% specificity, e.g. allowing 1:100 false-positive), sensitivities, i.e. true positives, of 30% and 61% was found for hepcidin and ERFE, respectively. For the ABP, the most sensitive marker was Off-hr score ([Hb] (g·L-1) - 60 × √RET%) (P < 0.05) with a maximal sensitivity of ~58% and ~ 9% following donation and reinfusion, respectively. Combining the findings for hepcidin, ERFE and the ABP yielded a sensitivity across all time-points of 83% following reinfusion in BT. Endurance performance increased 24 hours (+6.4%, P < 0.01) and six days following reinfusion (+5.8%, P < 0.01).Conclusions: Hepcidin and ERFE may serve as biomarkers in an anti-doping context following an ergogenic, small-volume blood transfusion.

Blood doping is prohibited for athletes but has been a well-described practice within endurance sports throughout the years. With improved direct and indirect detection methods, the practice has allegedly moved towards micro-dosing, that is, reducing the blood doping regime amplitude. This narrative review evaluates whether blood doping, specifically recombinant human erythropoietin (rhEpo) treatment and blood transfusions are performance-enhancing, the responsible mechanism as well as detection possibilities with a special emphasis on micro-dosing. In general, studies evaluating micro-doses of blood doping are limited. However, in randomized, double-blinded, placebo-controlled trials, three studies find that infusing as little as 130 ml red blood cells or injecting 9 IU × kg bw

Some have questioned the evidence for performance-enhancing effects of several substances included on the World Anti-Doping Agency's Prohibited List due to the divergent or inconclusive findings in randomized controlled trials (RCTs). However, inductive statistical inference based on RCTs-only may result in biased conclusions because of the scarcity of studies, inter-study heterogeneity, too few outcome events, or insufficient power. An abductive inference approach, where the body of evidence is evaluated beyond considerations of statistical significance, may serve as a tool to assess the plausibility of performance-enhancing effects of substances by also considering observations and facts not solely obtained from RCTs. Herein, we explored the applicability of an abductive inference approach as a tool to assess the performance-enhancing effects of substances included on the Prohibited List. We applied an abductive inference approach to make inferences on debated issues pertaining to the ergogenic effects of recombinant human erythropoietin (rHuEPO), beta2-agonists and anabolic androgenic steroids (AAS), and extended the approach to more controversial drug classes where RCTs are limited. We report that an abductive inference approach is a useful tool to assess the ergogenic effect of substances included on the Prohibited List - particularly for substances where inductive inference is inconclusive. Specifically, a systematic abductive inference approach can aid researchers in assessing the effects of doping substances, either by leading to suggestions of causal relationships or identifying the need for additional research.

Objectives Optimized concurrent training regimes are warranted in physical training of military-, law enforcement- and rescue-personnel. This study investigated if four 15-min endurance training sessions weekly improve aerobic capacity and performance more than one 60-min endurance session weekly during the initial phase of a Basic Military Training program. Design A randomized training intervention study with functional and physiological tests before and after the intervention. Methods Military conscripts (n = 290) were randomly allocated to three groups completing 9 weeks training. Weekly training consisted of four endurance and four strength training sessions lasting 15 min each (‘Micro-training’: MIC); one strength and one endurance session lasting 60 min each (‘Classical-training’: CLA) or two 60 min sessions of standard military training (‘Control-training’: CON). Results Both 12-min (∼7−10%) and shuttle run performance (∼35−42%) improved (P ≤ 0.001) similarly in all groups. Likewise, functional 2-min maximal repetition exercise capacity increased (P ≤ 0.05) similarly in all groups (Lunges ∼17−24 %; PushUp ∼10−20%; AbdominalFlexions∼21−23%). Peak oxygen uptake changes depended on group (P ≤ 0.05) with increases (P ≤ 0.01) in MIC (7 ± 7%, n = 23) and CON (12 ± 18%, n = 17) and no changes in CLA. Maximal m. vastus lateralis citrate synthase activity decreased 14 ± 26% (P ≤ 0.001, n = 18) in CLA. Likewise, maximal m. vastus lateralis 3-hydroxyacyl-CoA dehydrogenase activity decreased 8 ± 17% in MIC (n = 28) and 14 ± 24% in CLA (n = 18). Conclusions Four 15-min endurance training sessions weekly improves running performance and strength-endurance similarly to one 60 min session. Peak oxygen uptake only increases with more than one endurance session weekly and leg muscle oxidative capacity appears reduced after basic military training.

Objectives: To investigate whether recombinant human erythropoietin (rHuEPO) injections during an altitude training camp impact heart function. Methods: Thirty (12 women) moderately trained subjects stayed at 2320 m altitude for 4 weeks while training. Subjects were randomized to placebo (isotonic saline) or rHuEPO (20 IU/kg body weight) i.v. injections. Transthoracic echocardiography imaging was acquired 3 days after arrival to altitude and prior to the first placebo or rHuEPO injection as well as one day after the last rHuEPO injection three weeks later. Results: rHuEPO did not alter cardiovascular morphology parameters, systolic or diastolic function. In the placebo group, altitude exposure improved left ventricle (LV) systolic function due to an increased twist angle but rHuEPO had no additional effects. Pulmonary arterial systolic pressure was unaffected in either group. Notably, rHuEPO hampered LV untwist rate without affecting LV early filling. Conclusion: rHuEPO provided during mild altitude exposure does not cause any major effects on heart function. The observed alteration in LV untwist induced by rHuEPO is unlikely to have a meaningful clinical effect., World Anti-Doping Agency ISF17R02NN

Detailed physiological phenotyping was hypothesized to have predictive value for Olympic distance cross-country mountain bike (XCO-MTB) performance. Additionally, mean (MPO) and peak power output (PPO) in 4 × 30 s all-out sprinting separated by 1 min was hypothesized as a simple measure with predictive value for XCO-MTB performance. Parameters indicative of body composition, cardiovascular function, power and strength were determined and related to XCO-MTB national championship performance (n = 11). Multiple linear regression demonstrated 98% of the variance (P < 0.001) in XCO-MTB performance (tXCO-MTB; [min]) is explained by maximal oxygen uptake relative to body mass (VO2peak,rel; [ml/kg/min]), 30 s all-out fatigue resistance (FI; [%]) and with a minor contribution from quadriceps femoris maximal torque (Tmax; [Nm]): tXCO-MTB = -0.217× VO2peak,rel.-0.201× FI+ 0.012× Tmax+ 85.4. Parameters with no additional predictive value included hemoglobin mass, leg peak blood flow, femoral artery diameter, knee-extensor peak workload, jump height, quadriceps femoris maximal voluntary contraction force and rate of force development. Additionally, multiple linear regression demonstrated parameters obtained from 4x30s repeated sprinting explained 88% of XCO-MTB variance (P < 0.001) with tXCO-MTB = -5.7× MPO+ 5.0× PPO+ 55.9. In conclusion, XCO-MTB performance is predictable from VO2peak,rel and 30 s all-out fatigue resistance. Additionally, power variables from a repeated sprint test provides a cost-effective way of monitoring athletes XCO-MTB performance.

The novel hypothesis that "Live High-Train Low" (LHTL) does not improve sport-specific exercise performance (e.g., time trial) is discussed. Indeed, many studies demonstrate improved performance after LHTL but, unfortunately, control groups are often lacking, leaving open the possibility of training camp effects. Importantly, when control groups, blinding procedures, and strict scientific evaluation criteria are applied, LHTL has no detectable effect on performance.

Purpose: To investigate the hypothesis that a therapeutic oral dose of Tramadol improves cycling time trial performance and compromises motor-cognitive performance in highly trained cyclists.Methods: Following two familiarization trials, 16 highly trained cyclists completed a preloaded time trial (1 hour at 60 % of peak power followed by a 15 km time trial) after ingestion of 100 mg Tramadol or placebo in a double-blind placebo-controlled counterbalanced cross-over design separated by at least 4 days wash-out. Visuo-motor tracking and math tasks were completed during the preload (n=10) to evaluate effects on cognition and fine motor performance.Results: Time trial mean power output (298±42 W vs. 294±44 W) and performance (1474±77 s vs. 1483±85 s) were similar with Tramadol and placebo treatment, respectively. In addition, there were no differences in perceived exertion, reported pain, blood pH, lactate or HCO3 concentrations across trials. Heart rate was higher (PPConclusion: In highly trained cyclists, ingestion of 100 mg Tramadol does not improve performance in a 15 km cycling time trial that was completed after a one hour preload at 60 % peak power. Additionally, a therapeutic dose of Tramadol does not compromise complex motor-cognitive or simple fine motor performances.

Objectives Military-, rescue- and law-enforcement personnel require a high physical capacity including muscular strength. The present study hypothesized that 9 weeks of volume matched concurrent short frequent training sessions increases strength more efficiently than less frequent longer training sessions. Design A randomized training intervention study with functional and physiological tests before and after the intervention. Methods Military conscripts (n = 290) were assigned to micro-training (four 15-min strength and four 15-min endurance bouts weekly); classical-training (one 60-min strength and one 60-min endurance training session weekly) or a control-group (two 60-min standard military physical training sessions weekly). Results There were no group difference between micro-training and classical-training in measures of strength. Standing long jump remained similar while shotput performance was reduced (P ≤ 0.001) in all three groups. Pull-up performance increased (P ≤ 0.001) in micro-training (7.4 ± 4.6 vs. 8.5 ± 4.0 repetitions, n = 59) and classical-training (5.7 ± 4.1 vs. 7.1 ± 4.2 repetitions, n = 50). Knee extensor MVC increased (P ≤ 0.01) in all groups (micro-training, n = 30, 11.5 ± 8.9%; classical-training, n = 24, 8.3 ± 11.5% and control, n = 19, 7.5 ± 11.8%) while elbow flexor and hand grip MVC remained similar. Micro-training increased (P ≤ 0.05) type IIa percentage from 32.5 ± 11.0% to 37.6 ± 12.3% (n = 20) and control-group increased (P ≤ 0.01) type IIax from 4.4 ± 3.0% to 11.6 ± 7.9% (n = 8). In control-group type I, fiber size increased (P ≤ 0.05) from 5121 ± 959 μm to 6481 ± 2084 μm (n = 5). Satellite cell content remained similar in all groups. Conclusions Weekly distribution of low-volume concurrent training completed as either eight 15-min bouts or two 60-min sessions of which 50% was strength training did not impact strength gains in a real-world setting.

The time course of plasma volume (PV) reduction following an increased training load period is unknown and was investigated. The accompanying fluctuations in [Hb] and OFF-hr score were analyzed in the Athlete Biological Passport. Further, whether fluctuations in plasma albumin, soluble transferrin receptors (sTfR), and pro-atrial natriuretic peptide (proANP) concentrations correlate with PV fluctuations was investigated. Eleven high-level competitive cyclists were investigated for 3 weeks. After initial measurements in week 1, training load was increased ~250% in week 2 followed by a reversion to baseline training load in week 3. PV and hematological variables were determined frequently during all weeks. The higher training load in week 2 increased (P

Purpose: This study tested the hypothesis that autologous blood transfusion (ABT) of ~50% of the red blood cells (RBCs) from a standard 450 ml phlebotomy would increase mean power in a cycling time trial. Additionally, the study investigated whether further ABT of RBCs obtained from another 450 ml phlebotomy would increase repeated cycling sprint ability.Methods: In a randomized, double-blind, placebo-controlled crossover design (3-month wash-out), nine highly trained male subjects donated two 450 ml blood bags each (BT-trial) or were sham phlebotomized (PLA-trial). Four weeks later, a 650 kcal time trial (n=7) was performed three days before and 2 h after receiving either ~50% (135 ml) of the RBCs or a sham transfusion. On the following day, transfusion of RBCs (235 ml) from the second donation or sham transfusion was completed. A 4×30 s all-out cycling sprint interspersed by 4 min of recovery was performed six days before and three days after the second ABT (n=9).Results: The mean power was increased in time trials from before to after transfusion (PConclusion: ABT of only ~135 ml of RBCs is sufficient to increase mean power in a 650 kcal cycling time trial by ~5% in highly trained men. In contrast, a combined high-volume transfusion of ~135 and ~235 ml of RBCs does not alter 4×30 s all-out cycling performance interspersed with 4 min of recovery.