Post-injection hybridization of complementary DNA strands on capillary electrophoresis platforms: A novel solution for dsDNA artifacts

Several laboratories have reported the occurrence of a split or n − 1 peak at the vWA locus in PowerPlex^® 16 and PowerPlex^® ES amplification products separated on 4- and 16-capillary electrophoresis instruments. The root cause of this artifact is post-PCR reannealing of the unlabeled, unincorporated vWA primer to the 3′-end of the tetramethylrhodamine (TMR)-labeled strand of the vWA amplicon. This reannealing occurs in the capillary post-electrokinetic injection. The split peak is eliminated by incorporation into the loading cocktail of a sacrificial hybridization sequence (SHS) oligonucleotide that is complementary to the vWA primer. The SHS preferentially anneals to the primer instead of the TMR-labeled strand of the vWA amplicon. In addition, the n − 10/n − 18 artifact that may be seen at the vWA locus was determined to be due to double-stranded amplicon formed post-electrokinetic injection into the capillary. This was also eliminated by adding in two Complementary Oligo Targets (COT1 and COT2) in addition to the SHS oligonucleotide into the loading cocktail. These three oligonucleotides are complementary to the 33 bases at the 5′-end of the unlabeled vWA amplicon strand and the 60 bases at its 3′-end and therefore compete for hybridization to the TMR-labeled amplicon strand. Incorporation of these three oligonucleotides in the Internal Lane Standard 600 (ILS600) eliminate both the split peak and n − 10/n − 18 artifact in PowerPlex^® 16 and PowerPlex^® ES amplification products without affecting sizing of alleles at the vWA locus or any locus in the PowerPlex^® 16, PowerPlex^® Y, PowerPlex^® ES, AmpFlSTR^® Profiler Plus^® ID, AmpFlSTR^® Cofiler^®, and AmpFlSTR^® SGM Plus^® kits.     

Relationship of life events and social support with psychological dysfunction among adolescents

Relationships among major life events, perceived social support, and psychological disorder were assessed in a sample of older adolescents. Negative life events and satisfaction with social support were significantly and independently related to a range of psychological symptoms. Further, the relationship between negative events and disorder was moderated by gender, the types of events experienced, and anticipated change in the psychosocial environment. The importance of the use of standardized and psychometrically sound measures of life events, social support, and psychological disorder is highlighted.Received his Ph.D. from UCLA in 1980. An assistant professor, he is currently investigating stress and coping among children and adolescents.Received her Ph.D. from the University of Vermont in 1985. An assistant professor, she is currently studying social support.Received Ed. M. from Harvard Universtiy. He is a doctoral student in clinical psychology interested in the role of social support in coping with stress during adolescence.Received her BA from the University of Vermont. She is a doctoral student in clinical psychology.     

Fiscal rules in the Canadian provinces: Abject failure or qualified success?

Most Canadian provinces have introduced legislation to require politicians of all ideological stripes to meet annual balanced budget (BB) targets. Critics of this type of legislation argue that it is unnecessary, confining, and subject to manipulation. We examine provincial balanced budget legislation before and after the Great Recession and argue that the response of provinces needs to be evaluated taking into account the multiple objectives of fiscal policy and the behavioural changes that the rules introduce, including creative non‐compliance. Rules should be evaluated on their ability to contribute to good fiscal governance rather than on their stringency.     

GrinLine Identification Using Digital Imaging and Adobe Photoshop*

Abstract: The purpose of this study was to outline a method by which an antemortem photograph of a victim can be critically compared with a postmortem photograph in an effort to facilitate the identification process. Ten subjects, between 27 and 55 years old provided historical pictures of themselves exhibiting a broad smile showing anterior teeth to some extent (a grin). These photos were termed “antemortem” for the purpose of the study. A digital camera was used to take a current photo of each subject’s grin. These photos represented the “postmortem” images. A single subject’s “postmortem” photo set was randomly selected to be the “unknown victim.” These combined data of the unknown and the 10 antemortem subjects were digitally stored and, using Adobe Photoshop software, the images were sized and oriented for comparative analysis. The goal was to devise a technique that could facilitate the accurate determination of which “antemortem” subject was the “unknown.” The generation of antemortem digital overlays of the teeth visible in a grin and the comparison of those overlays to the images of the postmortem dentition is the foundation of the technique. The comparisons made using the GrinLine Identification Technique may assist medical examiners and coroners in making identifications or exclusions.