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.     

Interpretation of unusual wounds caused by firearms.

The interpretation of the range of fire of gunshot wounds requires coordination of information and observations from the autopsy surgeon, scene investigator, and laboratory analyst. Opinions based on incomplete information often lead to misinterpretation of the wound. The effects of interposed targets such as clothing, windows, and body parts are discussed. Case examples are given for interposed targets that cause confusing patterns and even lead to misidentification of the points of entrance and exit of the projectile in the body. The splitting of tissue in areas of bony prominence can be mistaken for evidence of discharge of a firearm at close range.     

The Tarasoff dilemma in criminal court.

The duty to protect, or Tarasoff duty, has been conceptualized as arising solely in the context of a clinical setting. A recent California Supreme Court ruling in People v. Clark adds legal, clinical, and ethical dilemmas to the oftentimes contentious Tarasoff issue. Though the Tarasoff issue is but a minor legal point in Clark, a possible consequence of Clark is that a Tarasoff warning could be deemed nonconfidential and admissible in a criminal trial. Psychotherapists could therefore be testifying in criminal courts as prosecution witnesses. While the possibility of a chilling effect on patients' disclosure of violent ideation in the context of psychotherapy first caused apprehension after the California Supreme Court's 1976 decision in Tarasoff v. Regents of the University of California, this same Court's ruling in People v. Clark some 14 years later may ensure that this fear finally becomes realized.     

Defining subnational open government: does local context influence policy and practice?

Policy Sciences - What is open government? The contemporary conceptualization of open government remains rooted in transparency and accountability, but it is embedded within the political economy...     

How Government Reactions to Violence Worsen Social Welfare: Evidence from Peru

Dissident violence inflicts many costs on society, but some of the longest‐lasting consequences for civilians may be indirect, due to the government's response. We explore how government policy responses affect social welfare, specifically through budgetary shifts. Using subnational violence and budgeting data for Peru, we demonstrate that attacks on soldiers during the budget negotiation period drive a shift from local social services, especially health, to defense. One soldier fatality implies a 0.13 percentage point reduction in the local health budget share (2008–12). Health budget cuts due to a single soldier fatality result in 76 predicted additional infant deaths 2 years later. We show that the effect on health budgeting operates through decreases in women's use of health facilities and postnatal services. We offer evidence that Peru's coercive response indirectly harms civilians due to butter‐to‐guns budgetary shifts. Our results identify a budgetary mechanism that translates dissident violence into a deterioration in social welfare.