The Downstream Benefits of Experimentation

The debate about the cost-effectiveness of randomized fieldexperimentation ignores one of the most important potentialuses of experimental data. This article defines and illustrates"downstream" experimental analysis—that is, analysis ofthe indirect effects of experimental interventions. We arguethat downstream analysis may be as valuable as conventionalanalysis, perhaps even more so in the case of laboratory experimentation.     

The use of SEM/EDS analysis to distinguish dental and osseus tissue from other materials

With increasing frequency, relatively small, fragmentary evidence thought to be osseous or dental tissue of human origin is submitted to the forensic laboratory for DNA analysis with the request for positive identification. Prior to performing DNA analysis, however, it is prudent to first perform a presumptive test or "screen" to determine whether the questioned material may be eliminated from further consideration. When material is shown not to be consistent with bone/teeth, DNA testing is not performed. When such determinations cannot be made from gross morphological features, elemental analysis can be indicative. This presumptive test is made possible by applying scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) in conjunction with an X-ray spectral database recently developed by the FBI laboratory. This database includes spectra for many different materials including known examples of bone and tooth from many different contexts and representing the full range of taphonomic conditions. Results of SEM/EDS analysis of evidence can be compared to these standards to determine if they are consistent with bone and/or tooth and, if not, then what the material might represent. Analysis suggests that although the proportions and amounts of calcium and phosphorus are particularly important in differentiating bone and tooth from other materials, other minor differences in spectral profile can also provide significant discrimination. Analysis enables bone and tooth to be successfully distinguished from other materials in most cases. Exceptions appear to be ivory, mineral apatite, and perhaps some types of corals.     

Validation of a 16-locus fluorescent multiplex system

STR multiplexes have been indispensable for the efficient genotyping of forensic samples. The PowerPlex 16 System contains the coreCODIS loci, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, CSF1PO, FGA, THOI, TPOX, vWA, the sex determinant locus, amelogenin, and two pentanucleotide STR loci, Penta D and Penta E. This multiplex satisfies the locus requirements for most national databases and is the most efficient currently available system due to its single PCR amplification. To provide the groundwork for judicial acceptance, including the publication of primer sequences, and to evaluate laboratory-to-laboratory variation, a developmental validation for casework on this commercially available system was performed in 24 laboratories and produced the following conclusions. Amplification was reliable on a variety of thermal cyclers and product could be analyzed on either an ABI PRISM 310 Genetic Analyzer or an ABI PRISM 377 DNA Sequencer. Genotyping using single source samples was consistent between 0.25 and 2 ng of input DNA template with a few laboratories obtaining complete genotypes at 0.0625 ng. However, heterozygote allele imbalance (<60% peak height balance) caused by stochastic effects was observed at a rate of 13% with 0.125 ng DNA and 22% at 0.0625 ng DNA. Mixture analyses were done using a total of 1 ng of DNA template. Most alleles were detected in mixtures of 4 to 1 and some minor alleles were detected in mixtures of 19 to 1. Optimum amplification cycle number was dependent on the sensitivity of the detection instrument used and could also be adjusted to accommodate larger amounts of DNA on solid supports such as FTA paper. Reaction conditions including volume, annealing temperature, and concentrations of primer, AmpliTaq Gold, and magnesium were shown to be optimal yet robust enough to withstand moderate variations without affecting genotype analysis. Environmental, matrix and standard source analyses revealed an ability to obtain complete genotypes in all sample types except those exposed to 80 degrees C for 12-48 days. Finally, comparison of genotype results from the PowerPlex 16 System with other commercially available systems on non-probative reference and forensic samples showed consistent results.     

An improved method for post-PCR purification for mtDNA sequence analysis

Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are desirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QlAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT reagent. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.     

Driver and front seat passenger fatalities associated with air bag deployment. Part 2: A review of injury patterns and investigative issues

Assessment of the role of air bag deployment in injury causation in a crash of any severity requires analysis of occupant, vehicle, and impact data. The potential injurious role of an air bag is independent of crash severity and is more obvious in minor collisions, particularly those involving "out-of-position" occupants. Factors such as occupant height and other constitutional and medical factors, intoxication, age, type, and proper use of other restraint systems, pre-impact braking and multiple impacts can contribute to an occupant being "out-of-position." Two injury mechanisms are described in out-of-position occupants: "punch-out" when the individual covers the air bag module before deployment and "membrane-force" when the occupant contacts a partly deployed air bag. Each mechanism is associated with injury patterns. In adults, "punch-out" can cause thoraco-abdominal trauma and "membrane-force" loading can lead to craniocervical injury. This can also occur in short-statured occupants including children subjected to both types of loading. In more severe collisions, other factors, e.g., intrusion, steering column and seatbelt loading and other occupant compartment contacts, can contribute to trauma.     

Population genetics of nine short tandem repeat loci: allele frequency distribution in a Brazilian population sample

Gene and genotype frequencies in relation to the D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, and D7S820 loci were determined in a sample of 290 unrelated individuals (204 Caucasians and 86 mulattoes) living in the city of S?o Paulo, Brazil. The sex test Amelogenin was also performed in all subjects from our sample, revealing the expected sex in all instances. Allele frequency data obtained from the analysis of these samples were in the usual range of other population groups with similar racial background. In the sample of Caucasian individuals, panmictic proportions were ruled out in relation to TPOX and CSF1PO loci, but only in the latter was the overall frequency of heterozygotes significantly less than expected. In the sample of mulattoes, Hardy-Weinberg proportions were rejected in relation to FGA and CSF1PO loci, but in no instance were the overall numbers of heterozygotes different from the corresponding expected ones under panmixia. Taking into account all this and also the number of tests performed, the degree of genetic heterogeneity of Brazilian populations, and the critical level reached by the significant results (1% < alpha<5%), the departures from panmixia here observed can be considered to be negligible in altering significantly biologic relationship odds calculated under the assumption of random matings.     

Understanding Children's Use of Secrecy in the Context of Eyewitness Reports

To investigate socioemotional influences on children's eyewitness accuracy, we examined children's reports for activities they were motivated to conceal. Forty-eight 3–6-year-old children participated in a standardized play session with their mothers. Half of the children were told by an experimenter not to play with certain toys, but did so at the urging of their mothers, who told their children to keep the play activities secret. The remaining children were not restricted from playing with the toys, nor told by their mothers to keep the play activities secret. Later, all children were interviewed about the activities with free narrative and detailed questions. Half were given an interview that consisted of highly suggestive questions; half were given an interview consisting of specific, less suggestive questions. Results indicated that older children who were instructed to keep events secret withheld more information than did older children not told to keep events secret. Younger children's reports were not significantly affected by the secret manipulation. There were no significant effects associated with interview type. We discuss implications for understanding the development of children's knowledge and use of secrecy, and applications of the research to issues that arise when child witnesses give reports in legal contexts.