William Goodwin

Demographic characteristics of elite Ethiopian endurance runners.

INTRODUCTION The dominance of East-African athletes in distance running remains largely unexplained; proposed reasons include favorable genetic endowment and optimal environmental conditions. PURPOSE To compare the demographics of elite Ethiopian athletes with the general Ethiopian population and assess the validity of reports linking running long distances to school with endurance success. METHODS Questionnaires, administered to 114 members (male and female) of the Ethiopian national athletics team and 111 Ethiopian control subjects (C) obtained information on place of birth, language, distance and method of travel to school. Athletes were separated into three groups according to athletic discipline: marathon (M; N = 34); 5,000-10,000 m (5-10 km; N = 42); and other track and field athletes (TF; N = 38). Frequency differences between groups were assessed using contingency chi-square tests. RESULTS Regional distributions of marathon athletes differed from controls (P < 0.001) and track and field athletes (P = 0.013), but not the 5- to 10-km athletes (P = 0.21). The 5- to 10-km athletes also differed from controls (P < 0.001). Marathon athletes exhibited excess from the regions of Arsi and Shewa (M: 73%; 5-10 km: 43%; TF: 29%; C: 15%). The language distribution of marathon athletes differed from all groups (P < 0.001), with a predominance of languages of Cushitic origin (M: 75%, 5-10 km: 52%, TF: 46%, C: 30%). A higher proportion of marathon athletes ran to school (M: 68%; 5-10 km: 31%; TF: 16%; C: 24%) and traveled greater distances. CONCLUSION Elite endurance athletes are of a distinct environmental background in terms of geographical distribution, ethnicity, and also having generally traveled farther to school, often by running. These findings may reflect both environmental and genetic influences on athletic success in Ethiopian endurance athletes.

Mitochondrial haplogroups associated with elite Kenyan athlete status.

UNLABELLED The maternal inheritance of mitochondrial DNA (mtDNA) has enabled construction of detailed phylogenies. Analysis of key polymorphisms from these phylogenies allows mtDNA to be assigned to haplogroups, which have been associated with elite endurance performance. PURPOSE To compare the frequencies of mtDNA haplogroups found in elite Kenyan athletes with those in the general Kenyan population. METHODS DNA samples were obtained from 221 national level Kenyan athletes (N), 70 international Kenyan athletes (I), and 85 members of the general Kenyan population (C). mtDNA haplogroups were classified by sequencing 340 bases of hypervariable section (HVS I) and by genotyping known restriction sites. Frequency differences between groups were assessed using exact tests of population differentiation. RESULTS The haplogroup distribution of national (P = 0.023) and international athletes (P < 0.001) differed significantly from controls, with international athletes showing a greater proportion of L0 haplogroups (C = 15%, N = 18%, I = 30%) and lower proportion of L3* haplogroups (C = 48%, N = 36%, I = 26%). Although a high number of international athletes originated from the Rift Valley province relative to controls (C = 20%, N = 65%, I = 81%), subjects from this province did not differ in haplogroup distribution from other regions (P = 0.23). Nor did Bantu subjects differ from Nilotic (P = 0.12) despite an overrepresentation of Nilotic languages among the athletes. CONCLUSIONS International athletes differed in their mtDNA haplogroup distribution relative to the general Kenyan population. They displayed an excess of L0 haplogroups and a dearth of L3* haplogroups. These findings suggest that mtDNA haplogroups are influential in elite Kenyan distance running, although population stratification cannot be ruled out.

Y chromosome haplogroups of elite Ethiopian endurance runners

Favourable genetic endowment has been proposed as part of the explanation for the success of East African endurance athletes, but no evidence has yet been presented. The Y chromosome haplogroup distribution of elite Ethiopian athletes (n=62) was compared with that of the general Ethiopian population (n=95) and a control group from Arsi (a region producing a disproportionate number of athletes; n=85). Athletes belonged to three groups: marathon runners (M; n=23), 5–km to 10–km runners (5–10K; n=21) and other track and field athletes (TF; n=18). DNA was extracted from buccal swabs and haplogroups were assigned after the typing of binary markers in multiplexed minisequencing reactions. Frequency differences between groups were assessed by using contingency exact tests and showed that Y chromosome haplogroups are not distributed amongst elite Ethiopian endurance runners in the same proportions as in the general population, with statistically significant (P<0.05) differences being found in four of the individual haplogroups. The geographical origins and languages of the athletes and controls suggest that these differences are less likely to be a reflection of population structure and that Y chromosome haplogroups may play a significant role in determining Ethiopian endurance running success.

A novel multiplex assay for simultaneously analysing 13 rapidly mutating Y-STRs

A multiplex polymerase chain reaction (PCR) assay (RM-Yplex) was developed which is capable of simultaneously amplifying 13 recently introduced rapidly mutating Y-STR markers (RM Y-STRs). This multiplex assay is expected to aid human identity testing in forensic and other applications to improve differentiating unrelated males and allow separating related males. The 13 RM Y-STR markers included in the multiplex are: DYF387S1, DYF399S1, DYF403S1ab, DYF404S1, DYS449, DYS518, DYS526ab, DYS547, DYS570, DYS576, DYS612, DYS626 and DYS627. This study reflects the proof of concept to analyse all currently known RM Y-STRs simultaneously and describes the optimization of the multiplex assay. The RM-Yplex assay generated complete RM Y-STR profiles down to 62.5pg of male template DNA, and from male-female DNA mixtures at all ratios tested. We herewith introduce and make available for widespread use in forensic and anthropological studies, an effective and sensitive single multiplex assay for simultaneous genotyping of 13 RM Y-STRs.

STR data for the AmpFℓSTR® Identifiler® loci in Kuwaiti population

Allele frequencies for 15 STR loci included in AmpFlSTR Identifiler kit were ascertained in a sample population of 502 unrelated Kuwaiti individuals. Allele frequencies were compared with 6 Caucasian populations using an exact test. This showed that the Kuwaiti population was very similar to the neighboring Iraqi and Saudi populations. As the geographical distance between the populations increased, as expected, more differences were observed. Relevant forensic parameters were also determined.

Development of internal amplification controls for DNA profiling with the AmpFℓSTR(®) SGM Plus(®) kit.

DNA extracted from forensic samples can be degraded and also contain co‐extracted contaminants that inhibit PCR. The effects of DNA degradation and PCR inhibition are often indistinguishable when examining a DNA profile. Two internal amplification controls (IACs) were developed to improve quality control of PCR using the AmpFℓSTR® SGM Plus® kit. The co‐amplification of these controls with DNA samples was used to monitor amplification efficiency and detect PCR inhibitors. IAC fragments of 90 and 410 bp (IAC90 and IAC410) were generated from the plasmid pBR322 using tailed primers and then amplified with ROX‐labelled primers. Co‐amplification of IAC90 and IAC410 was performed with varying amounts of template DNA, degraded DNA and DNA contaminated with humic acid, heme and indigo dye. Both IAC90 and IAC410 were successfully amplified with human DNA without significantly affecting the quality of the DNA profile, even with DNA amounts lower than 0.5 ng. In the presence of inhibitors, the IAC90 signal was still present after all human DNA loci fail to amplify; in contrast, the IAC410 signal was reduced or absent at low levels of inhibition. Amplification of the two IACs provided an internal PCR control and allowed partial profiles caused by inhibition to be distinguished from degraded DNA profiles.

DNA transfer: The role of temperature and drying time

It has previously been shown, and reconfirmed here, that biological material on a substrate will transfer readily upon contact with another substrate when wet but hardly when dry. There is however a paucity of data regarding the speed at which body fluids dry and how this may affect its transfer upon contact. Here we conduct transfer experiments at 4°C, 22°C and 40°C at multiple time points during the drying process. The speed at which blood dries is dependent on the temperature, with the drying process complete within 15-60min. The percentage of deposited DNA transferred upon contact follows an exponential pattern of decline from soon after deposition, decreasing until the sample is dry. There are no differences in transfer rates upon contact among the different temperature conditions within the first 5min or after 60min since deposit, but significant variation occurs between these time points. When considering the likelihood of a proposed scenario that incorporates one or more contact situations it is important to consider the timing of the potential transfer event(s) relative to when the biological sample in question was initially deposited. The results of this study will assist the interpretation and evaluation of alternative scenarios involving transfer of biological substances.

Concordance between the AmpFlSTR MiniFiler and AmpFlSTR Identifiler PCR amplification kits in the Kuwaiti population.

Abstract:  The AmpFℓSTR® MiniFiler™ polymerase chain reaction amplification kit, developed and supplied by Applied Biosystems, complements the AmpFℓSTR® Identifiler® polymerase chain reaction amplification kit (Applied Biosystems, Warrington, U.K.) by improving the success rate when profiling DNA that is degraded or contains inhibitors. Before applying the MiniFiler™ kit to casework, the profiles from 200 unrelated Kuwaitis were compared to Identifiler® profiles. Concordance was observed for 99.875% (1598 of 1600) of the compared STR loci. The two discordant profiles displayed allelic dropout: one at the D13S317 locus due to nonamplification of allele 10 in the MiniFiler™ profile, and one at the D18S51 locus due to nonamplification of allele 18 in the Identifiler® profile.

Development of a multiplex system to assess DNA persistence in taphonomic studies

In this study, we have developed a PCR multiplex that can be used to assess DNA degradation and at the same time monitor for inhibition: primers have been designed to amplify human, pig, and rabbit DNA, allowing pig and rabbit to be used as experimental models for taphonomic research, but also enabling studies on human DNA persistence in forensic evidence. Internal amplified controls have been added to monitor for inhibition, allowing the effects of degradation and inhibition to be differentiated. Sequence data for single‐copy nuclear recombination activation gene (RAG‐1) from human, pig, and rabbit were aligned to identify conserved regions and primers were designed that targeted amplicons of 70, 194, 305, and 384 bp. Robust amplification in all three species was possible using as little as 0.3 ng of template DNA. These have been combined with primers that will amplify a bacterial DNA template within the PCR. The multiplex has been evaluated in a series of experiments to gain more knowledge of DNA persistence in soft tissues, which can be important when assessing what material to collect following events such as mass disasters or conflict, when muscle or bone material can be used to aid with the identification of human remains. The experiments used pigs as a model species. When whole pig bodies were exposed to the environment in Northwest England, DNA in muscle tissue persisted for over 24 days in the summer and over 77 days in the winter, with full profiles generated from these samples. In addition to time, accumulated degree days (ADD) were also used as a measure that combines both time and temperature—24 days was in summer equivalent to 295 ADD whereas 77 days in winter was equivalent to 494 ADD.

Theoretical value of the recommended expanded European Standard Set of STR loci for the identification of human remains

We have undertaken a series of simulations to assess the effectiveness of commercially available sets of STR loci, including the loci recommended for inclusion in the expanded European Standard Set, for the purpose of human identification. A total of 9200 genotype simulations were performed using DNA · VIEW. The software was used to calculate likelihood ratios (LRs) for 23 groups of relatives, and to determine the probability of identification given scenarios that ranged between 10 and 250,000 victims. The additional loci included in the recommended expanded European Standard Set, when used in conjunction with the Identifiler® kit, significantly improved the typical LRs for tested scenarios and the likely success of providing correct identifications.