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.     

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...     

Eurosceptic Challenger Parties and Political Opposition in European Union Politics: Part of the Problem or Part of the Solution?

Despite extensive research on Eurosceptic challenger parties, our knowledge of their influence on political opposition has so far been sparse. In this article we make an in‐depth assessment of parliamentary EU opposition, based on 4,264 statements made by national parliamentarians in the European Affairs Committees (EACs) of Denmark and Sweden. Our analysis shows that the presence of Eurosceptic challenger parties in the national parliamentary arena impacts patterns and practices of EU opposition significantly. A greater presence of ‘hard’ Eurosceptic parties in parliament is associated with more opposition in EU politics. These parties deliver a vast majority of the polity‐oriented opposition towards the EU and present more policy alternatives than mainstream parties. The findings presented have implications for our understanding of national parliamentary EU opposition as well as for the assessment of the impact of Eurosceptic challenger parties on the process of European integration.     

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.     

The Balkan experience: Explanations of public affairs and interest group activity

This final article is a synthesis of findings from the seven country studies in this special issue. Although there are characteristics common to all group systems, particularly those transitioning to democracy, there are also significant differences. Analysis of these differences reveals a direct relationship between both the level of socioeconomic and political development and the characteristics of the group systems in all Balkan countries. In addition, we provide new insights into group system development and operation. These are particularly related to the impact of war, inter‐ethnic cleavages, constitutional arrangements (majoritarian and consociational), economic crises, geopolitics, and the major presence of external interests.