Application of low copy number STR typing to the identification of aged, degraded skeletal remains

Low copy number (LCN) STR typing was successfully applied to four interesting cases during developmental validation of the approach for degraded skeletal remains. Specific questions were addressed in each case, with the acquisition of STR data largely serving as additional confirmatory or investigatory information in any specific situation, and not necessarily providing the definitive evidence to establish identity. The cases involve missing U.S. service members from World War I, World War II, and the Vietnam War. The variety of these cases, in terms of the questions addressed, the age of the remains, and the type of reference material available for comparison, demonstrates the broad utility of LCN STR typing in the identification of degraded skeletal remains from missing persons.     

Forensic typing of autosomal SNPs with a 29 SNP-multiplex—Results of a collaborative EDNAP exercise

We report the results of an inter-laboratory exercise on typing of autosomal single nucleotide polymorphisms (SNP) for forensic genetic investigations in crime cases. The European DNA Profiling Group (EDNAP), a working group under the International Society for Forensic Genetics (ISFG), organised the exercise. A total of 11 European and one US forensic genetic laboratories tested a subset of a 52 SNP-multiplex PCR kit developed by the SNPforID consortium. The 52 SNP-multiplex kit amplifies 52 DNA fragments with 52 autosomal SNP loci in one multiplex PCR. The 52 SNPs are detected in two separate single base extension (SBE) multiplex reactions with 29 and 23 SNPs, respectively, using SNaPshot kit, capillary electrophoresis and multicolour fluorescence detection. For practical reasons, only the 29 SBE multiplex reaction was carried out by the participating laboratories. A total of 11 bloodstains on FTA cards including a sample of poor quality and a negative control were sent to the laboratories together with the essential reagents for the initial multiplex PCR and the multiplex SBE reaction. The total SNP locus dropout rate was 2.8% and more than 50% of the dropouts were observed with the poor quality sample. The overall rate of discrepant SNP allele assignments was 2.0%. Two laboratories reported 60% of all the discrepancies. Two laboratories reported all 29 SNP alleles in all 10 positive samples correctly. The results of the collaborative exercise were surprisingly good and demonstrate that SNP typing with SBE, capillary electrophoresis and multicolour detection methods can be developed for forensic genetics.     

What doctors and policymakers should know: Battered women's views about mandatory medical reporting laws

Medical professionals in forty-five states and the District of Colombia are under a legal obligation to report a variety of injuries to the police, including those believed to be the result of domestic violence. Responses to these laws by doctors and battered women have been mixed. Using logistic regression analysis, this study explored battered women's views about mandatory medical reporting laws controlling for and examining a variety of demographic, situational, and legal factors. Three of the four dependent attitudinal measures (perceptions of general support for mandatory reporting as well as perceived benefits of the law for themselves and others) demonstrated little variation across all factors. The fourth measure - willingness to seek medical care in communities with a mandatory law - however, produced a number of significant relationships among the demographic, situational and legal factors. The results and policy implications of these findings are discussed.     

Insights from Zambian miners for rethinking development policy and the environment

The world is at a critical point as humanity contemplates how its own activity is contributing to changes in the earth and atmosphere. Formidable challenges require raising fundamental questions and learning from unlikely sources. Drawing on field research conducted on the Zambian Copperbelt, this article explores how public conversations concerning differing views of reality can inform development-related thinking about the environment. Enumerating practical examples where words and images both conveyed and shaped conflicting viewpoints in the industrial mine setting, the article asserts that much can be learned from the experiential viewpoints of underground miners. Policy making could benefit, for instance, from lessening its dependence on dominant economic thinking and increasingly drawing upon historical, cultural, philosophical, and theological insights when devising policies, projects, and procedures. Questions of power, control, and humanity's self-conception in relation to the physical world are also explored.     

INTERNATIONAL HEALTH CARE REFORM: THE NEED FOR COMPREHENSIVE MICRO-COMPARATIVE ANALYSIS

Microcomparison, or single‐component analysis, of health care systems offers a potentially better basis for reform than traditional macrocomparison analysis of aggregate elements. Using macroanalysis, available evidence shows that Germany provides cheaper but more effective hospital care than the United States. To find the causes for this outcome, we developed a microanalytic model of hospital administrators’ perceptions, financial ratios, medical outcomes, and pharmaceutical costs. However, only data on pharmaceutical costs were available and similar in both countries. Our significant outcome was development of a microcomparative model that gives world medical care providers new criteria for analyzing and improving cost to care rafios.     

Making Connections: Performance Regimes and Extreme Events

Local officials in the emergency management field have reached out and increased their connections with other agencies and organizations during the past several years. Collaborative networks have been created in an effort to address the complexities and uncertainties surrounding extreme events. But has this collaboration really taken root? In this article, the authors find that although a collaborative ethos has penetrated local emergency management, it is neither deep nor uniform. Data from a survey of emergency managers in North Carolina counties show that maintaining a functional network—a performance regime in which participants develop consistent management practices and rely on each other for the generation of new ideas—is a difficult task. The explanation for the variation found across the counties largely involves capacity and vulnerability.     

Improved MtDNA sequence analysis of forensic remains using a "mini-primer set" amplification strategy

Mitochondrial DNA (mtDNA) analysis of highly degraded skeletal remains is often used for forensic identification due largely to the high genome copy number per cell. Literature from the "ancient DNA" field has shown that highly degraded samples contain populations of intact DNA molecules that are severely restricted in size (1-4). Hand et al. have demonstrated the targeting and preferential amplification of authentic human DNA sequences with small amplicon products of 150 bp or less (1,2). Given this understanding of ancient DNA preservation and amplification, we report an improved approach to forensic mtDNA analysis of hypervariable regions 1 and 2 (HV1/HV2) in highly degraded specimens. This "mini-primer set" (MPS) amplification strategy consists of four overlapping products that span each of the HV regions and range from 126 to 170 bp, with an average size of 141 bp. For this study, 11 extracts representing a range of sample quality were prepared from nonprobative forensic specimens. We demonstrate a significant increase in MPS amplification success when compared to testing methods using approximately 250 bp amplicons. Further, 16 of 17 independent amplifications previously "unreported" due to mixed sequences provided potentially reportable sequence data from a single, authentic template with MPS testing.     

The Use of Mitochondrial DNA Single Nucleotide Polymorphisms to Assist in the Resolution of Three Challenging Forensic Cases

Abstract:  Mitochondrial DNA (mtDNA) single nucleotide polymorphisms (SNPs) in an 11-plex assay were typed in three missing person cases involving highly degraded human remains. Unlike the traditional forensic approach to analyzing mtDNA which focuses on sequencing portions of the noncoding Control Region, this assay targets discriminatory SNPs that reside principally in the coding region. In two of the cases, the SNP typing successfully excluded one of two reference families that could not be excluded on the basis of mtDNA hypervariable region sequencing alone, and resulted in the final resolution of both decades-old cases. In a third case, SNP typing confirmed the sorting and reassociation of multiple commingled skeletal elements. The application of a specific mtDNA SNP assay in these cases demonstrates its utility in distinguishing samples when the most common Caucasian hypervariable region type is encountered in forensic casework.     

Wheels within Wheels: Poverty,Power and Patronage in the Cambodian Migration System

Abstract

This article presents evidence for an adjusted and refocused systems theory of labour migration in Cambodia. Specifically, it seeks to highlight first, how migration in Cambodia may be understood as a multi-scalar phenomenon characterised by pragmatism and flexibility; secondly, it emphasises the undergirding role of traditional rural norms in shaping and mediating the systematic process of labour movement; and finally, it presents evidence concerning how these structures constitute a vessel of social change, not only from urban to rural, but also from the rural to urban. In this way, a picture is presented of Cambodian migration as an adaptable, but nevertheless highly patterned process which is rapidly reordering the Kingdom’s cities and villages alike.     

Assessing a novel room temperature DNA storage medium for forensic biological samples

The ability to properly collect, analyze and preserve biological stains is important to preserving the integrity of forensic evidence. Stabilization of intact biological evidence in cells and the DNA extracts from them is particularly important since testing is generally not performed immediately following collection. Furthermore, retesting of stored DNA samples may be needed in casework for replicate testing, confirmation of results, and to accommodate future testing with new technologies.A novel room temperature DNA storage medium, SampleMatrix™ (SM; Biomatrica, Inc., San Diego, CA), was evaluated for stabilizing and protecting samples. Human genomic DNA samples at varying amounts (0.0625-200 ng) were stored dry in SM for 1 day to 1 year under varying conditions that included a typical ambient laboratory environment and also through successive freeze-thaw cycles (3 cycles). In addition, spiking of 1-4× SM into samples prior to analysis was performed to determine any inhibitory effects of SM. Quantification of recovered DNA following storage was determined by quantitative PCR or by agarose gel electrophoresis, and evaluation of quantitative peak height results from multiplex short tandem repeat (STR) analyses were performed to assess the efficacy of SM for preserving DNA.Results indicate no substantial differences between the quality of samples stored frozen in liquid and those samples maintained dry at ambient temperatures protected in SM. For long-term storage and the storage of low concentration samples, SM provided a significant advantage over freezer storage through higher DNA recovery. No detectable inhibition of amplification was observed at the recommended SM concentration and complete profiles were obtained from genomic DNA samples even in the presence of higher than recommended concentrations of the SM storage medium. The ability to stabilize and protect DNA from degradation at ambient temperatures for extended time periods could have tremendous impact in simplifying and improving sample storage conditions and requirements. The current work focuses on forensics analysis; however this technology is applicable to all endeavors requiring storage of DNA.