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The Science of Sex Verification and Athletic Performance

Ross Tucker and Malcolm Collins

The division of athletes into male and female categories for competition is a widely accepted practice and is ordinarily straightforward, requiring no intervention from authorities. However, for reasons ranging from deliberate cheating to complex medical conditions resulting in ambiguous development of sex organs, the controversy of sex verification in athletic events has existed for 70 years. Testing procedures, initially implemented to prevent cheating by men masquerading as women, have produced humiliating outcomes for women athletes who were often for the first time confronted with the possibility that they have one of the disorders of sex development. Sporting authorities have and continue to formulate position stands for the management of such cases. An important missing component in this debate is the sound scientific evidence to determine (a) whether a performance advantage actually exists and (b) how large it might be. The division of competition into separate categories and the large difference in sporting performance between male and female necessitate that sport-governing bodies define the boundaries between the sexes in a just and fair manner for all participating athletes. This review will therefore provide the historical context of the debate and aim to discuss relevant physiological and performance aspects of the sex verification process.

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A Polymorphism in a Functional Region of the COL5A1 Gene: Association With Ultraendurance-Running Performance and Joint Range of Motion

Shameemah Abrahams, Michael Posthumus, and Malcolm Collins


Endurance-running performance and joint range of motion (ROM) are both multifactorial phenotypes. A single-nucleotide polymorphism, rs172722 (C/T), in the COL5A1 3′-untranslated region (UTR) was shown to independently associate with both phenotypes. Two major functional forms of the COL5A1 3′-UTR have been identified and differ by 7 tightly linked polymorphisms that include rs12722 and a short tandem-repeat polymorphism (STRP rs71746744, –/AGGG). It has been proposed that STRP rs71746744 plays a role in the predicted secondary structures and mRNA stability of the 2 major forms of the COL5A1 3′-UTR, therefore implying a regulatory role. The aim of this study was to determine whether STRP rs71746744 is independently associated with running performance and prerace sit-and-reach range of motion (SR ROM) in a cohort of ultramarathon road runners.


One hundred six (74 men and 32 women, age 22–67 y) white runners who participated in either the 2009 or 2011 Two Oceans 56-km ultramarathon were included in this cross-sectional study. Their SR ROM measurements, COL5A1 rs71746744 genotype, and overall race times were determined.


COL5A1 rs71746744 was independently associated with running performance (P = .024) and prerace sr rom (P = .020). Moreover, the AGGG/AGGG genotype was significantly overrepresented in the fastest and inflexible athletes compared with those with either the –/AGGG or –/– genotype.


These findings provide further evidence for a relationship between COL5A1, running performance, and SR ROM. Further studies are needed to investigate the effect of this variant on the mechanical properties of connective tissue.

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Caffeine Ingestion Does Not Alter Performance during a 100-km Cycling Time-Trial Performance

Angus M. Hunter, Allan St, Clair Gibson, Malcolm Collins, Mike Lambert, and Timothy D. Noakes

This study analyzed the effect of caffeine ingestion on performance during a repeated-measures, 100-km, laboratory cycling time trial that included bouts of 1- and 4-km high intensity epochs (HIE). Eight highly trained cyclists participated in 3 separate trials—placebo ingestion before exercise with a placebo carbohydrate solution and placebo tablets during exercise (Pl), or placebo ingestion before exercise with a 7% carbohydrate drink and placebo tablets during exercise (Cho), or caffeine tablet ingestion before and during exercise with 7% carbohydrate (Caf). Placebo (twice) or 6 mg · kg−1 caffeine was ingested 60 min prior to starting 1 of the 3 cycling trials, during which subjects ingested either additional placebos or a caffeine maintenance dose of 0.33 mg · kg−1 every 15 min to trial completion. The 100-km time trial consisted of five 1-km HIE after 10, 32, 52, 72, and 99 km, as well as four 4-km HIE after 20, 40, 60, and 80 km. Subjects were instructed to complete the time trial and all HIE as fast as possible. Plasma (caffeine) was significantly higher during Caf (0.43 ± 0.56 and 1.11 ± 1.78 mM pre vs. post Pl; and 47.32 ± 12.01 and 72.43 ± 29.08 mM pre vs. post Caf). Average power, HIE time to completion, and 100-km time to completion were not different between trials. Mean heart rates during both the 1-km HIE (184.0 ± 9.8 Caf; 177.0 ± 5.8 Pl; 177.4 ± 8.9 Cho) and 4-km HIE (181.7 ± 5.7 Caf; 174.3 ± 7.2 Pl; 175.6 ± 7.6 Cho; p < .05) was higher in Caf than in the other groups. No significant differences were found between groups for either EMG amplitude (IEMG) or mean power frequency spectrum (MPFS). IEMG activity and performance were not different between groups but were both higher in the 1-km HIE, indicating the absence of peripheral fatigue and the presence of a centrally-regulated pacing strategy that is not altered by caffeine ingestion. Caffeine may be without ergogenic benefit during endurance exercise in which the athlete begins exercise with a defined, predetermined goal measured as speed or distance.

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The Interaction of Aging and 10 Years of Racing on Ultraendurance Running Performance

Dale E. Rae, Andrew N. Bosch, Malcolm Collins, and Mike I. Lambert

The aim of this study was to examine the interaction between aging and 10 years of racing in endurance runners. Race-time data from 194 runners who had completed 10 consecutive 56-km ultramarathons were obtained. The runners were either 20.5 ± 0.7, 30.0 ± 1.0, 39.9 ± 0.9, or 49.4 ± 1.0 years old at their first race. Each runner’s race speed was determined for each race over the 10 years. Data were analyzed using repeated-measures ANOVA, one-way ANOVA, and independent t tests and showed that performance improved and declined at greater rates for younger runners; younger runners had a greater capacity for improvement than older runners; ≈4 years were required to reach peak racing speed, regardless of age; it was not possible to compete at peak speed for more than a few years; and the combined effects of 10 years of aging and racing neither improve nor worsen net performance. In conclusion, these data suggest that although these runners showed similar patterns of change in race speed over a 10-year period, the extent of change in performance was greater in younger than in older runners.

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The COL5A1 Gene, Ultra-Marathon Running Performance, and Range of Motion

James C. Brown, Caron-Jayne Miller, Michael Posthumus, Martin P. Schwellnus, and Malcolm Collins


Endurance running performance is a multifactorial phenotype that is strongly associated with running economy. Sit and reach range of motion (SR ROM) is negatively associated with running economy, suggesting that reduced SR ROM is advantageous for endurance running performance. The COL5A1 gene has been associated with both endurance running performance and SR ROM in separate cohorts. The aim of this study was to investigate whether COL5A1 is associated with ultra-marathon running performance and whether this relationship could be partly explained by prerace SR ROM.


Seventy-two runners (52 male, 20 female) were recruited from the 56 km Two Oceans ultra-marathon and were assessed for prerace SR ROM. The cohort was genotyped for the COL5A1 BsfUI restriction fragment length polymorphism, and race times were collected after the event.


Participants with a TT genotype (341 ± 41 min, N = 21) completed the 56 km Two Oceans ultra-marathon significantly (P = 0.014) faster than participants with TC and CC genotypes (365 ± 39 min, N = 50). The COL5A1 genotype and age accounted for 19% of performance variance. When the cohort was divided into performance and flexibility quadrants, the T allele was significantly (P = 0.044) over-represented within the fast and inflexible quadrant.


The COL5A1 genotype was found to be significantly associated with performance in a 56 km ultra-endurance run. This study confirms previous findings and it furthers our understanding of the relationships among ROM, COL5A1, and endurance running performance. We continue to speculate that the COL5A1 gene alters muscle-tendon stiffness.

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Genetic Polymorphisms Related to VO2max Adaptation Are Associated With Elite Rugby Union Status and Competitive Marathon Performance

Elliott C.R. Hall, Sandro S. Almeida, Shane M. Heffernan, Sarah J. Lockey, Adam J. Herbert, Peter Callus, Stephen H. Day, Charles R. Pedlar, Courtney Kipps, Malcolm Collins, Yannis P. Pitsiladis, Mark A. Bennett, Liam P. Kilduff, Georgina K. Stebbings, Robert M. Erskine, and Alun G. Williams

Purpose: Genetic polymorphisms have been associated with the adaptation to training in maximal oxygen uptake ( V ˙ O 2 max ). However, the genotype distribution of selected polymorphisms in athletic cohorts is unknown, with their influence on performance characteristics also undetermined. This study investigated whether the genotype distributions of 3 polymorphisms previously associated with V ˙ O 2 max training adaptation are associated with elite athlete status and performance characteristics in runners and rugby athletes, competitors for whom aerobic metabolism is important. Methods: Genomic DNA was collected from 732 men including 165 long-distance runners, 212 elite rugby union athletes, and 355 nonathletes. Genotype and allele frequencies of PRDM1 rs10499043 C/T, GRIN3A rs1535628 G/A, and KCNH8 rs4973706 T/C were compared between athletes and nonathletes. Personal-best marathon times in runners, as well as in-game performance variables and playing position, of rugby athletes were analyzed according to genotype. Results: Runners with PRDM1 T alleles recorded marathon times ∼3 minutes faster than CC homozygotes (02:27:55 [00:07:32] h vs 02:31:03 [00:08:24] h, P = .023). Rugby athletes had 1.57 times greater odds of possessing the KCNH8 TT genotype than nonathletes (65.5% vs 54.7%, χ 2 = 6.494, P = .013). No other associations were identified. Conclusions: This study is the first to demonstrate that polymorphisms previously associated with V ˙ O 2 max training adaptations in nonathletes are also associated with marathon performance (PRDM1) and elite rugby union status (KCNH8). The genotypes and alleles previously associated with superior endurance-training adaptation appear to be advantageous in long-distance running and achieving elite status in rugby union.