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.     

PMS,psychosis and culpability: sound or misguided defense?

Premenstrual Syndrome (PMS) is believed to affect up to 90% of women of reproductive age. A small subset of women have been identified who actually experience psychotic symptoms in the premenstrual phase of their cycles. Not surprisingly, PMS has made it into the courts where it has been offered as a defense for criminal acts. The defense has generally fared poorly in the United States, although it has been successfully used as a factor in supporting diminished capacity in Great Britain. The following paper sets out to review the medical literature on premenstrual syndrome with a particular focus on premenstrual psychosis. Available literature from both medical and legal sources is then utilized to investigate instances in which premenstrual psychosis has been invoked as a defense in the courts.     

Crystallization of cerebrospinal fluid in cases of death from ischemic heart disease and ethanol poisoning

Crystallographic analysis of the cerebrospinal fluid (CSF) was carried out in 18 cases of death from coronary disease and 19 cases of death from ethanol poisoning. The crystallograms were evaluated visually and by the stereoscopic picture. Specific features of the CSF crystal colonies growth in subjects dead from the above conditions are determined.     

A new improved method for extraction of DNA from teeth for the analysis of hypervariable loci

A new method for better recovery of DNA suitable for amplification of hypervariable loci from fragments of teeth, consisting of two steps-scraping and aspiration, and extensive decalcification-is reported. Higher yields of high molecular weight DNA were obtained from the root, pulp, and crown of all kinds of 120 teeth, irrespective of gender, age, and source of teeth. HLA DQA1, 5 poly markers (LDLR, GYPA, HBGG, D7S8, and Gc), and other 12 short tandem repeat loci (HPRTB, F13B, LPL, D13S317, D7S820, D5S818, D21S11, D18S51, FGA, D8S1179, D3S1358, and vWA) could be successfully amplified and typed from recovered DNA.     

Estimation of body exposure to explosion

An ordnance-disposal expert was killed while disposing of a cache of explosives. The likely position of the body was reconstructed by modeling the explosion as an omnidirectional emission of particles from a model of the explosion site and noting the distribution of particles on a model of a human. The applications and limitations of this method in reconstructing the events and correlation with the injuries noted at autopsy are discussed.     

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.