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Rock A.,University of Innsbruck | Irwin J.,413 Research Blvd | Dur A.,University of Innsbruck | Parsons T.,International Commission on Missing Persons | Parson W.,Innsbruck Medical University
Forensic Science International: Genetics | Year: 2011

The analysis of the haploid mitochondrial (mt) genome has numerous applications in forensic and population genetics, as well as in disease studies. Although mtDNA haplotypes are usually determined by sequencing, they are rarely reported as a nucleotide string. Traditionally they are presented in a difference-coded position-based format relative to the corrected version of the first sequenced mtDNA. This convention requires recommendations for standardized sequence alignment that is known to vary between scientific disciplines, even between laboratories. As a consequence, database searches that are vital for the interpretation of mtDNA data can suffer from biased results when query and database haplotypes are annotated differently. In the forensic context that would usually lead to underestimation of the absolute and relative frequencies. To address this issue we introduce SAM, a string-based search algorithm that converts query and database sequences to position-free nucleotide strings and thus eliminates the possibility that identical sequences will be missed in a database query. The mere application of a BLAST algorithm would not be a sufficient remedy as it uses a heuristic approach and does not address properties specific to mtDNA, such as phylogenetically stable but also rapidly evolving insertion and deletion events. The software presented here provides additional flexibility to incorporate phylogenetic data, site-specific mutation rates, and other biologically relevant information that would refine the interpretation of mitochondrial DNA data. The manuscript is accompanied by freeware and example data sets that can be used to evaluate the new software (http://stringvalidation.org). © 2010 Elsevier Ireland Ltd.

Parson W.,Innsbruck Medical University | Parson W.,Pennsylvania State University | Gusmao L.,State University of Rio de Janeiro | Gusmao L.,University of Porto | And 9 more authors.
Forensic Science International: Genetics | Year: 2014

The DNA Commission of the International Society of Forensic Genetics (ISFG) regularly publishes guidelines and recommendations concerning the application of DNA polymorphisms to the question of human identification. Previous recommendations published in 2000 addressed the analysis and interpretation of mitochondrial DNA (mtDNA) in forensic casework. While the foundations set forth in the earlier recommendations still apply, new approaches to the quality control, alignment and nomenclature of mitochondrial sequences, as well as the establishment of mtDNA reference population databases, have been developed. Here, we describe these developments and discuss their application to both mtDNA casework and mtDNA reference population databasing applications. While the generation of mtDNA for forensic casework has always been guided by specific standards, it is now well-established that data of the same quality are required for the mtDNA reference population data used to assess the statistical weight of the evidence. As a result, we introduce guidelines regarding sequence generation, as well as quality control measures based on the known worldwide mtDNA phylogeny, that can be applied to ensure the highest quality population data possible. For both casework and reference population databasing applications, the alignment and nomenclature of haplotypes is revised here and the phylogenetic alignment proffered as acceptable standard. In addition, the interpretation of heteroplasmy in the forensic context is updated, and the utility of alignment-free database searches for unbiased probability estimates is highlighted. Finally, we discuss statistical issues and define minimal standards for mtDNA database searches. © 2014 Elsevier Ireland Ltd.

Amory S.,International Commission on Missing Persons | Huel R.,International Commission on Missing Persons | Bilic A.,International Commission on Missing Persons | Loreille O.,413 Research Blvd | Parsons T.J.,International Commission on Missing Persons
Forensic Science International: Genetics | Year: 2012

During the 7 year period from 2002 to 2009 a high volume, silica-binding DNA extraction protocol for bone, based on modified QIAGEN's Blood Maxi Kit protocol was highly successful permitting the DNA matching of >14,500 missing persons from former Yugoslavia. This method, however, requires large amount of bone material and large volumes of reagents. The logical evolution was to develop a more efficient extraction protocol for bone samples that uses significantly less starting material while increasing the success in obtaining DNA results from smaller, more challenging samples. In this study we compared the performance of ICMP's original protocol against an automatable full demineralization approach. In order to provide reliable results and to simulate a wide variety of cases, we analyzed 40 bone samples in a comparative study based on DNA concentrations and quality of resulting STR profiles. The new protocol results in the dissolution of the entire bone powder sample, thus eliminating the possibility that DNA is left behind, locked in remaining solid bone matrix. For the majority of samples tested, the DNA concentrations obtained from half a gram of fully digested bone material were equivalent to or greater than the ones obtained from 2 g of partially demineralized bone powder. Furthermore, the full demineralization process significantly increases the proportion of full profiles reflecting the correlation with better DNA quality. This method has been adapted for the QIAcube robotic platform. The performance of this automated full demineralization protocol is similar to the manual version and increases overall lab throughput. It also simplifies the process by eliminating quality control procedures that are advisable in manual procedures, and overall reduces the chance of human error. Finally we described a simple and efficient post-extraction clean-up method that can be applied to DNA extracts obtained from different protocols. This protocol has also been adjusted for the QIAcube platform. © 2011 Elsevier Ireland Ltd. All rights reserved.

Irwin J.A.,413 Research Blvd | Saunier J.,413 Research Blvd | Bodner M.,Innsbruck Medical University | Amory S.,International Commission on Missing Persons | And 5 more authors.
International Journal of Legal Medicine | Year: 2010

In order to better characterize and understand the mtDNA population genetics of Central Asia, the mtDNA control regions of over 1,500 individuals from Uzbekistan have been sequenced. Although all samples were obtained from individuals residing in Uzbekistan, individuals with direct ancestry from neighboring Central Asian countries are included. Individuals of Uzbek ancestry represent five distinct geographic regions of Uzbekistan: Fergana, Karakalpakstan, Khorezm, Qashkadarya, and Tashkent. Individuals with direct ancestry in nearby countries originate from Kazakhstan, Kyrgyzstan, Russia, Afghanistan, Turkmenistan, and Tajikistan. Our data reinforce the evidence of distinct clinal patterns that have been described among Central Asian populations with classical, mtDNA, and Y-chromosomal markers. Our data also reveal hallmarks of recent demographic events. Despite their current close geographic proximity, the populations with ancestry in neighboring countries show little sign of admixture and retain the primary mtDNA patterns of their source populations. The genetic distances and haplogroup distributions among the ethnic populations are more indicative of a broad east-west cline among their source populations than of their relatively small geographic distances from one another in Uzbekistan. Given the significant mtDNA heterogeneity detected, our results emphasize the need for heightened caution in the forensic interpretation of mtDNA data in regions as historically rich and genetically diverse as Central Asia. © Springer-Verlag 2010.

Zukowski L.A.,University of Florida | Falsetti A.B.,International Commission on Missing Persons | Tillman M.D.,University of Florida
Journal of Anatomy | Year: 2012

Previous research on lumbar spine osteophyte formation has focused on patterned development and the relation of age and sex to degeneration within the vertebral bodies. The inclusion of osteophytes originating on the laminae and body mass index (BMI) may result in a more complete evaluation. This study investigates lumbar osteophyte development on the laminae and vertebral bodies to determine whether osteophyte development: (i) is related bilaterally, at different lumbar levels, and superior and inferior margins; (ii) on the laminae and vertebral bodies are reciprocally dependent responses; (iii) is correlated with sex, age and/or BMI. Seventy-six individuals (39 females, 37 males) were randomly selected from a modern skeletal collection (Bass Donated Collection). Osteophyte development was scored in eight regions on each vertebra at all five lumbar levels. A factor analysis considered all 40 scoring regions and Pearson's correlation analyses assessed the relatedness of age and BMI with the consequent factors. The factor analysis separated the variables into two similar factors for males and females defined as: (i) superior and inferior vertebral body scores and (ii) superior laminar scores at higher lumbar levels. The factor analysis also determined a third factor for females defined as: (iii) inferior laminar scores at lower lumbar levels. The severity of vertebral body osteophytes increased with age for both sexes. Additionally for females, as BMI increased, osteophyte severity increased for both the superior laminar margins higher in the column and the vertebral bodies. Dissimilarities between the factors in males and females and the correlation of BMI to osteophyte severity exclusively in females provide evidence for different biomechanical processes influencing osteophyte development. © 2011 The Authors. Journal of Anatomy © 2011 Anatomical Society of Great Britain and Ireland.

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