Search

Touch DNA analysis has become an important aspect of a forensic laboratory’s workload and a crucial tool for investigators in many cases. However, there is a lack of research regarding the influence of environmental conditions on Touch DNA, which is proven to reduce traces of biological material in samples. This study investigated the influence of time between deposition and recovery of Touch DNA, as well as the impact of temperature and humidity on a range of porous and non-porous surfaces.

There are different variables that affect the success of Touch DNA recovery, including surface type, the collection method used and extraction techniques. This experiment investigated how a range of porous and non-porous surfaces, different DNA collection (cotton swab, nylon flocked swab and SceneSafe Fast™ minitape) and extraction methods (PrepFiler Express BTA™ and QIAamp® DNA Investigator) affected touch DNA recovery.

DNA isolation in sexual assault cases is complicated by the presence of large numbers of female epithelial cells, which are often in vast excess when compared to spermatozoa. The two-step differential extraction has been a standard method for isolating the spermatozoa; however, new techniques in forensic science allow the use of precision techniques for capturing spermatozoa. We have used an immuno-magnetic bead-based technique for sperm cell separation. We identified two antibodies that were specific to spermatozoa: SP17 polyclonal antibody and SP10 Intra Acrosomal Protein monoclonal antibody. These were conjugated to Dynabeads® M-450 Epoxy beads and used to isolate the spermatozoa in samples that exhibited the characteristic of sexual assault samples. Using these antibodies, we achieved separation of spermatozoa in the samples with sperm concentration 104/ml and 103/ml; STR analysis produced full male profiles. Mixed profiles were obtained with reduced levels of spermatozoa. Our preliminary data illustrate the potential to apply antibody-based capture to sexual assault cases.

Short tandem repeat (STR) profiling has been routinely used in kinship testing since the introduction of commercial kits in the mid-1990s. While 15 to 23 STR loci normally give definitive results in simple kinship testing, additional loci are sometimes required to resolve complex cases. The SureID 23comp Human Identification Kit, recently released by Health Gene Technologies (China), multiplexes amelogenin and 22 autosomal STRs, 17 of which are non-CODIS STRs. This enables the profiling of 38–40 loci when used in conjunction with widely used commercial kits. In this study, the kit was evaluated for kinship applications as a supplementary STR kit following the minimum criteria for validation recommended by the European Network of Forensic Science Institutes (ENFSI) and the Scientific Working Group on DNA Analysis Methods (SWGDAM) using 500 samples. Performance was comparable with other commercial kits demonstrating: repeatability and reproducibility; precision (maximum s.d. 0.1048 nt); accuracy, all alleles were within ±0.41 nt compared to the actual sizes; heterozygous peak balances at all loci >68%; stutter ratios ranged from 3.8% to 16.15%; full profiles were generated with 125 pg DNA (95.12% of alleles at 62 pg),; and we found 100% concordance over 5 common STRs with the GlobalFiler kit.

Touch DNA, commonly known as trace DNA, is widely used in forensic DNA casework. However, touch DNA samples are challenging because of the many variables that can impact the success of obtaining a good quality usable DNA profile. Cotton and nylon flocked swabs are often used to collect touch DNA from surfaces, so this study aimed to test different techniques using cotton (150C) and nylon flocked (4N6FLOQSwabs®) swabs to collect touch DNA from non-porous surfaces. There was a significant difference amongst the three recovery techniques tested to recover touch DNA with cotton swabs and nylon swabs from textured plastic (p < 0.001), with a nylon swab and 30μl of distilled water being more efficient than a cotton swab with 100μl of distilled water. There was also a significant difference between the four recovery techniques to recover touch DNA from glass surfaces exposed to high humidity and low temperature (5 °C/78%) (p

The effective search for the missing and identification of persons, alive or dead, are core components in the prevention and in resolving the issue of Missing Persons. Despite the growing literature on this topic, there is still a lack of publications describing the Search as a process that includes different phases inherently composed of forensic investigative and identification principles for both living and deceased missing persons. This paper is the result of discussions between the Forensic Unit of the International Committee of the Red Cross (ICRC) and members of its external Forensic Advisory Board. It aims to present the Search process as an overarching concept that includes the investigation and identification phases of the missing in any state (dead or alive), in any scenario (with or without bodies), with an integrated, multidisciplinary, and multiagency approach for implementation by all actors involved in the investigation and identification phases of missing persons. [Abstract copyright: © 2021 The Authors.]

The effectiveness of five methods for the preservation of bone were assessed: cell lysis solution (with 1% sodium azide), dehydration/freeze-drying, ethanol (96%), freezing, and room temperature storage. Preserved bone samples were extracted using five optimised extraction methods. These preservation methods were tested for their efficiency for storage of fresh and degraded bone samples for 6 weeks, 6 months and 1 year. Freezing was found to be the best preservation method for longer-term storage of bone samples; this was followed by ethanol (96%), dehydration/freeze-drying, and room temperature storage. Full profiles were obtained from bone samples using all these preservation methods.

Mitochondrial DNA analysis has earned its place in the field of genetics for providing a new window into understanding population ancestry. Its ability to produce results from minimal or decayed samples where nucleus DNA profiling proves are difficult is an additional benefit to forensic genetics. A total of 150 whole blood samples were collected on FTA paper, from Arabs population in United Arab Emirates, including inhabitants from rural regions. Both extraction of whole blood samples using PrepFiler® as well as direct amplification of mtDNA control region from purified 1.2 mm disc of FTA card stained with blood were attempted in this study. Quantity of the amplified control region was estimated using gel electrophoresis. Three sets of primers were used afterward to sequence purified products of amplified control region of mtDNA using Sanger sequencing method. 150 mtGenome sequences were obtained for UAE Arabs population, generated using MPS technology – Ion™ 5S. Concordance with control region sequences obtained using Sanger Sequencing approach was investigated. Resulted haplotypes were compared against worldwide mtDNA database (EMPOP) and estimated haplotypes frequency is shown in this study. As a forensic lab, non-probative challenging bone samples were tested to highlight the potentials value of using MPS technology – Ion™ 5S. This study shows the first mtGenome data generated for UAE Arabs population which helps extending the current available mtDNA control region database. As a result, this study shows great value of implementing MPS in the routine work in forensic genetics at Dubai Police.

A set of 87 reference samples collected from the population of Saudi Arabia were sequenced using the ForenSeq™DNA Signature Prep Kit on a MiSeq FGx™. The FASTQ files contain the sequences of the SE33 STR, but are not reported by the ForenSeq™ Universal Analysis Software (UAS). The STRait Razor software was used to recover and to report SE33 sequence‐based data for the Saudi population. Ninety-six sequence-based alleles were recovered, most of which had previously reported motif patterns. Two unreported motif patterns found in three alleles and seven novel allele sequences were reported. We also reported a single discordance between the sequence-based data and the CE data that was due to the presence of a common TTTT deletion. SE33 had 130% more sequence-based alleles; the highest number of observed sequence variants were in alleles 27.2 and 30.2, which each had 7 sequence variants. The statistical parameters emphasize the usefulness of using the sequence-based data.

The SNPforID consortium identified a panel of 52 SNPs for forensic analysis that has been used by several laboratories worldwide. The original analysis of the 52 SNPs was based on a single multiplex reaction followed by two single-base-extension (SBE) reactions each of which was analyzed using capillary electrophoresis. The SBE assays were designed for high throughput genetic analyzers and were difficult to use on the single capillary ABI PRISM 310 Genetic Analyzer and the latest generation 3500 Genetic Analyzer, as sensitivity on the 310 was low and separation of products on the 3500 with POP-7™ was poor. We have modified the original assay and split it into four multiplex reactions, each followed by an SBE assay. These multiplex assays were analyzed using polymer POP-4™ on ABI 310 PRISM® and polymers POP-4™, POP-6™ and POP-7™ on the 3500 Genetic Analyzer. The assays were sensitive and reproducible with input DNA as low as 60 pg using both the ABI 310 and 3500. In addition, we found that POP-6™ was most effective with the 3500, based on the parameters that we assessed, achieving better separation of the small SBE products; this conflicted with the recommended use of POP-7™ by the instrument manufacturer. To support the use of the SNP panel in casework in Malaysia we have created an allele frequency database from 325 individuals, representing the major population groups within Malaysia. Population and forensic parameters were estimated for all populations and its efficacy evaluated using 51 forensic samples from challenging casework.

Blowfly species of the family Calliphoridae can be used in forensic investigations to estimate the minimum post-mortem interval (PMI¬min). Lucilia caesar and Lucilia illustris (Diptera: Calliphoridae) are closely related and phenotypically similar, making reliable identification difficult, especially if specimens are in poor condition. To identify potential markers to genetically distinguish these species five complete mitochondrial genomes were sequenced: three for L. caesar (KM657111- KM657113) and two mitochondrial genomes for L. illustris (KM657109, KM 657110). The ND6 gene contained the most species-specific SNPs (1.71%), followed by the ND5 gene (1.68%) and then the COI gene (1.56%), identifying ND6 and ND5 as valuable loci for differentiating L. Caesar and L. illustris specimens.

Touch DNA profiling is an important tool to solve the mystery of many cases, especially when other biological evidences cannot be found in crime scene. However, there are many variables that influence Touch DNA profiling such as recovery techniques and extraction. In addition, effect of environmental factors on items found outdoor such as sand can impact on the process. Therefore the aim of this experiment was to test how sandy surfaces can affect the recovery of Touch DNA Profiling by validation two recovery methods and two extraction kits that are widely used in the DNA forensic field.

Touch samples by their nature do not contain large amounts of biological material, especially compared to biological fluids such as blood, semen or saliva. Consequently, more careful recovery and storage are desirable in order to maximise the amount of DNA available. The main aim of the work presented is to determine whether detergent-based wetting agents increased DNA yield from touch samples when compared to water as a wetting agent. The results show that the use of the detergent-based lysis buffer led to greater DNA recovery from the fingerprints than when distilled water was used. The use of the lysis buffer in the touch DNA sample collection improved DNA recovery and stability over 24 h post-collection.

The effect of decalcification on pulverised bone samples was studied to evaluate its value when processing reasonably good quality samples. The decalcified and non-decalcified bone samples were extracted using phenol-chloroform extraction method, quantitated with GoTaq ® qPCR Master Mix (Promega ® , UK) using Applied Biosystems ® 7500 Real-Time PCR System (Thermo Fisher Scientific ® , UK) and amplified using an in-house amplification multiplex. The results showed that the decalcification of bone samples is not a necessary when processing good quality samples and higher DNA yields were obtained without the decalcification process. The electropherograms produced from the extracted DNA samples without decalcification were good quality and displayed no PCR inhibition.

The commercial PowerPlex® Fusion kit is an autosomal STR multiplex kit that has high discrimination power and is more informative in forensic, paternity, and relationship-testing cases. Key features of this multiplex system are the possibility to direct amplify FTA™ card punches as well as non-FTA cards and commonly used swabs; optimised inhibitor tolerance and high sensitivity generating full profiles from amount as little as 100 pg of human DNA. This study focused on the optimization of performance variables such as FTA™ punch sizes, reduced reaction volumes, and FTA™ purification reagent aiming to increase the analytical sensitivity, decrease the sample consumption, and cost effectiveness. LOD and LOQ values demonstrated high sensitivity of the PowerPlex® Fusion system. In addition, population databases of Brunei Malay and Chinese from the Brunei Darussalam were established, and parameters of forensic importance were calculated. Overall, the forensic parameters indicated an enhanced utility of the PowerPlex® Fusion kit for forensic evidence analysis and paternity testing in Brunei Malay and Chinese populations.

When profiling a reference dataset of 500 DNA samples for the population of Saudi Arabia, using the GlobalFiler® PCR amplification kit, six unusual alleles were detected. At the SE33 locus, four novel alleles were found: 2, 14.3, 20.3, and 38; two alleles, at the D1S1656 locus: 7 and 8, had been previously reported, but no published sequence data was available. The D1S1656 alleles were sequenced using ForenSeq™ DNA Signature Prep with the MiSeq FGx System (Illumina, USA). As the SE33 is not reported by available Massively Parallel Sequencing (MPS) systems, samples that exhibited the unreported alleles were sequenced using BigDye™ Terminator v3.1 Cycle Sequencing Kit. Here we present the sequence and structure of the previously uncharacterized alleles.

The Precision ID Ancestry panel from ThermoFisher contains 165 SNP markers. In this study we compared library construction using the automated workflow with the Ion Chef and a manual workflow. Casework samples that were collected between 2005 and 2018 in the State of Qatar where used for the study and the samples varied from routine stains to challenging samples that yielded full and partial STR profiles. One hundred and forty-three samples were processed: sixteen using both workflows. Full and partial profiles were seen with both methods. The assay was sensitive; one sample with only 0.12 ng of input DNA having a full profile. No significant differences were seen between the two workflows. However, the automated workflow saved considerable hands-on lab time and reduced the potential for pipetting errors.

This study profiled 1047 volunteers using the GlobalFiler™ Kit from the four main ethnic groups in Ghana: the Akan (282), Mole-Dagomba and Northern ethnic minority (253), Ewe (250) and Ga-Dangbe (262), representing about 98.4% ethnic coverage of the Ghanaian population. The polymorphic nature of the African population was demonstrated during the profiling of the reference dataset with the occurrence of some rare triplet variant alleles. At the TPOX locus, three tri-allelic variants were found with the allelic patterns 6-8-10, 8-9-10 and 9-10-11. In addition, tri-allelic patterns were seen at D5S818 (11-13-14) and D3S1358 (15-16-17). Although the TPOX tri-allelic pattern has been reported in other populations including African, the D5S818; 11-13-14 has never been reported in an African population. It has however, been reported once with a frequency of 1 in 69,600 in a convicted offender case in the USA. The D3S1358; 15-16-17 pattern has not been reported in an African population. In conclusion, the occurrence of rare alleles is illustrative of the genetic diversity within the Ghanaian population. This was further illustrated by the presence of three individuals that were heterozygous at all 21 STR loci.

Highlights:\ud * GlobalFiler® PCR amplification kit was used to generate a reference dataset for the population of Saudi Arabia.\ud * SE33, D12S391, and D1S1656 in this kit are more informative for the population of Saudi Arabia than any locus in the AmpFlSTR® Identifiler® PCR amplification kit.

Differentiating male lineages using non-recombining Y-chromosomal genetic markers is highly informative for tracing human migration and for forensic studies. Recently, it has been shown that the level of male lineage resolution can be enhanced by analysing rapidly mutating (RM) Y-STRs. The aim of this study was to develop an allelic frequency database for Qatari population to evaluate the resolution power of 13 RM Y-STRs. The overall haplotype diversity (HD) was 100%. It was found that the markers which contributed the most toward high HD were DYF399S1 and DYF403S1a/b. Together with their value for paternal male relative differentiation, these RM Y-STRs will be a valuable asset for forensic casework. AMOVA test was performed between Qatari population in comparison to Gulf countries, Middle East, and several worldwide population data sets. FST values were also calculated. Geography was found to account considerably for the pattern of population sub structuring. The RM Y-STR markers showed remarkable haplotype resolution power in the Qatari population, high gene diversity and sufficient robustness for a diverse range of applications.

The main focus in forensic genetics for two decades has been to improve the extraction of DNA from a wide variety of evidence and to make the profiling technology more sensitive and robust. In contrast, the recovery methods for biological material have seen little development. This study aims to improve the efficacy of the collection and storage processes, from crime scene to receipt at the laboratory. This study compared the use of ultrapure water as a wetting agent when collecting biological evidence using swabs with a detergent-based buffer. The results show that the stability post-collection greatly improved by using a newly developed buffer. When ultrapure water is used, DNA degradation was seen after 6 h at room temperature. However, the detergent-based buffer stabilized DNA for up to 48 h, even when the temperature was increased to 50 °C. The impact of these findings may be limited where crime scene evidence can be refrigerated until it reaches the laboratory. However, there are many situations/contexts where sample refrigeration is not possible and there is scope to improve the preservation of the genetic forensic evidence before it reaches the laboratory.

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.

DNA analysis was first applied to the identification of victims of armed conflicts and other situations of violence (ACOSV) in the mid-1990s, starting in South America and the Balkans. Argentina was the first country to establish a genetic database specifically developed to identify disappeared children. Following on from these programs the early 2000s marked major programs, using a largely DNA-led approach, identifying missing persons in the Balkans and following the attack on the World Trade Center in New York. These two identification programs significantly expanded the magnitude of events to which DNA analysis was used to help provide the identity of missing persons. Guidelines developed by Interpol (2014) [1] related to best practice for identification of human remains following DVI type scenarios have been widely disseminated around the forensic community; in numerous cases these guidelines have been adopted or incorporated into national guidelines/standards/practice. However, given the complexity of many humanitarian contexts in which forensic science is employed there is a lack of internationally accepted guidelines, related to these contexts, for authorities to reference. In response the Argentine government's Human Rights Division in the Ministry of Foreign Affairs and Worship (MREC) proposed that the United Nations (UN) should promote best practice in the use of forensic genetics in humanitarian forensic action: this was adopted by the UN in Resolutions A/HRC/RES/10/26 and A/HRC/RES/15/5. Following on from the adoption of the resolutions MREC has coordinated, with the support of the International Committee of the Red Cross (ICRC), the drafting of a set of guidelines (MREC, ICRC, 2014) [2], with input from national and international agencies. To date the guidelines have been presented to South America's MERCOSUR and the UN and have been disseminated to interested parties.

Biological samples recovered for forensic investigations are often degraded and/or have low amounts of DNA; in addition, in some instances the samples may be contaminated with chemicals that can act as PCR inhibitors. As a consequence this can make interpretation of the results challenging with the possibility of having partial profiles and false negative results. Because of the impact of DNA analysis on forensic investigations, it is important to monitor the process of DNA profiling, in particular the amplification reaction. In this chapter we describe a method for the in-house generation and use of internal amplification controls (IACs) with DNA profiling kits to monitor the success of the PCR proces. In the example we show the use of the SGM Plus® kit. These controls can also be used to aid the interpretation of the DNA profile.

Mitochondrial DNA (mtDNA) has been used in forensic investigations for over 20 years and continues to be employed at times when conventional short tandem repeat typing is not possible. The key features of mtDNA that make it valuable in forensic investigations are its high copy number and maternal mode of inheritance. Another feature of mtDNA is that the distribution of different mutations across the global populations is not uniform, thereby allowing geographical ancestry of biological evidence to be inferred, which can be critical in forensic investigations. Finally, mtDNA can also be a valuable tool for species identification.

Forensic DNA profiling uses a series of commercial kits that co-amplify several loci in one reaction; the products of the PCR are fluorescently labelled and analysed using CE. Before CE, an aliquot of the PCR is mixed with formamide and an internal lane size standard. Using the SGM Plus amplification kit, we have developed two internal non-amplified controls of 80 bp and 380 bp that are labelled with ROX fluorescent dye and added to the PCR. Combined with two internal amplification controls of 90 bp and 410 bp, they provide additional controls for the PCR, electrokinetic injection, and CE and also function as an internal size standard.

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.

(2012). Chapter 14 - Evidence for the ‘natural’ east African athlete. Routledge Online Studies on the Olympic and Paralympic Games: Vol. 1, East African Running: toward a cross-disciplinary perspective, pp. 257-281.

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.

Using the Applied Biosystems' AmpFlSTR SGM Plus PCR amplification kit, we studied the allele frequency distribution of 10 STR loci in two south Asian populations: one from the Gujarat region of India represented by 172 unrelated Gujaratis, now resident in England; and a Pakistani population, represented by 155 unrelated individuals. Gujarat borders southeast Pakistan. There were no significant deviations from Hardy-Weinberg equilibrium in either population after Bonferroni correction. The combined power of discrimination and exclusion for the Indian population were 0.999999999999544 and 0.9999785, respectively; for the Pakistani population, they were 0.999999999999865 and 0.9998975, respectively. F(ST) (or theta) between these two populations was estimated as 0.00146.

After death DNA becomes progressively more fragmented as biological tissue degrades and this results in a decreasing ability to gain a complete STR profile. When extracting and profiling DNA from human remains understanding the likely persistence of DNA in different tissues is important. Studies in the UK have demonstrated that when using pigs as an experimental model, DNA up to 400bp will persist for up to 3 weeks in the summer. However, it is well known that DNA degradation, especially in muscle tissue that has not become dehydrated, is dependent to a large degree on temperature. To assess DNA persistence in more extreme environmental conditions pig carcasses were exposed to the environment in Thailand during June for 10 days, with samples being collected every 12h: muscle tissue was present for up to three days post-mortem. Extracted DNA could not be amplified after 36h exposure unless the muscle was collected from tissue that was in contact with the ground; DNA persisted for up to 72h in these samples.

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 62.5 pg 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. (C) 2015 Elsevier Ireland Ltd. All rights reserved.

The mtDNA and Y chromosomes of the Malay population of peninsular Malaysia show high levels of diversity. Both the Malay and the indigenous Orang Asli populations showed a similar level of divergence between the different Y chromosome haplotypes.

East African athletes now dominate international distance running events from the 800 m to the marathon. Explanations for their phenomenal success have included optimal environmental conditions for developing distance running performance, psychological advantage and advantageous physiological characteristics. It is well established that genetics plays a role in determining inter-individual differences in exercise performance and adaptation to training stimuli. It is not known, however, to what extent inter-population differences (i.e. between ‘races’ and/or ethnic groups) in exercise performance can be attributed to genetics. There have been considerations that ‘black’ athletes are genetically adapted towards performance, given the concurrent success of athletes of West African ancestry in sprint events. However, the current notion of ‘race’ is not universally accepted, and genetic differences within and between populations are not clearly delineated by geographical or ethnic categorizations. Recent findings from mitochondrial DNA show that the populations from which Ethiopian athletes are drawn have not been isolated populations and are not genetically distinct from other Ethiopians. Y-chromosome analysis of the same population shows concurrent results, although some differences are present between athletes and the general Ethiopian population, suggesting an influence of the Y chromosome on athlete status in Ethiopia. It is concluded that there may be a role for genetics in the success of East African athletes; however, any genetic component to their success is unlikely to be limited to East Africans and is more likely to be found in other populations. At present it is unjustified to implicate a role for genetics in the success of East African runners when no genes have been identified as being important to their performance.

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

INTRODUCTION: Folate is a methylating agent which can reduce the aggressiveness of gliomas by remethylating genes involved in proliferation and apoptosis. Folate is a potential therapeutic treatment for cancer. It can be found naturally as Folinic Acid, or in the synthetic form Folic Acid, and can cross the blood brain barrier. Methylation is a useful target as it is an epigenetic change that can be reversed. METHOD: To begin to investigate the effect of folate on glioma, glioma cell lines U87 and 1321, plus the glial cell line SVG were used. Glioma cell lines were treated for 7 days with Folic and Folinic Acid at 4 and 40 µg/ml with and without an antioxidant. Cell viability was assessed using the PrestoBlue assay and cell cycle and apoptosis were tested using Propidium Iodide and caspase Glo 3/7 respectively. RESULTS: Cells treated with the higher level of Folinic Acid, 40 µg/ml, showed significant reduction in cell viability compared with the control (P < 0.01). Cell cycle analysis showed no difference in the percentages of each different cell cycle phase between the control and treatments on either Day 2 or Day 7. This suggests that the reduction in cell viability may be due to changes in apoptosis. CONCLUSION: These findings suggest that folate has an effect on cell viability in glioma cell lines. Folate could also have a beneficial effect on gliomas, which supports the use of folate for further research on primary glioma tissue.

Chromosome configurations and chiasma frequency during the metaphase I stage of spermatogenesis in the male horse are characterized in this work. The genome-wide frequency and distribution of chiasmata was detected as 49.45 ± 2.07 for 14 fertile stallions. All X and Y chromosomes shared a single chiasma at their pseudoautosomal region, while 1-4 chiasmata were observed in autosomal chromosomes. The chiasma frequency and distribution were further studied for 8 different bivalents identified by FISH in 5 fertile stallions. Genetic length was calculated from chiasmata data for the whole genome as well as for these 8 chromosomes. The findings complement the genetic linkage data and provide insight into the genetic basis of spermatogenesis in normal stallions.

In order to demonstrate the sequence diversity of mitochondrial D-loop DNA in the Taiwanese Han population, we established a database of 155 unrelated individuals. For each individual, the complete 980bp DNA region from the 5' end of HVI to 3' end of HVII segment was sequenced. In these 155 sequence data, 149 different haplotypes were observed, amongst these haplotypes, 144 were unique, 4 were found in 2 individuals and 1 was found in 3 individuals. When compare to the Anderson sequence, 144 transitions, 24 transversions, 5 insertions and 5 deletions were found. Eight positions exhibited more than one polymorphic sequence, six exhibited two variants while two exhibited three variants. Over the 1024bp that was analysed, pairwise differences between the sequences were 11.35+/-3.53bp. The sequence and nucleotide diversity were 0.9994 and 0.0116, respectively. The probability of two individuals randomly matching over the entire control region was 0.007. The diversity in the mitochondrial D-loop indicates the value of this locus for casework within Taiwan.