A systematic approach to improve downstream single-cell analysis for the DEPArray™ technology

Most commercially available STR amplification kits have never been fully validated for low template DNA analysis, highlighting the need for testing different PCR kits and conditions for improving single-cell profiling. Here, current strategies rely mainly on adjusting PCR cycle number and analytical threshold settings, with a strong preference for using 30 amplification cycles and thresholds at 30–150 RFU for allele detection. This study aimed to (1) determine appropriate conditions for obtaining informative profiles utilizing a dilution series, and (2) test the outcome on single cells using the DEPArray™ technology. Four routinely applied forensic STR kits were compared by using three different amplification volumes and DNA dilutions down to 3.0 pg, while two well-performing kits were used for single/pooled leucocyte and sperm cell genotyping. Besides reduced costs, the results demonstrate that a 50%–75% PCR volume reduction was beneficial for peak height evaluation. However, this was counteracted by an increased artifact generation in diluted DNA volumes. Regarding profile completeness, the advantage of volume reduction was only prominent in samples processed with Fusion 6C. For single and pooled cells, ESIFast and NGMDetect provided a solid basis for consensus profiling regarding locus failure, although locus dropouts were generally observed as stochastic events. Amplification volume of 12.5 μL was confirmed as appropriate in terms of peak heights and stutter frequencies, with increased stutter peaks being the main artifact in single-cell profiles. Limitations associated with these analyses are discussed, providing a solid foundation for further studies on low template DNA.     

States and their international relations since 1816: introducing version 2 of the International System(s) Dataset (ISD)

ABSTRACT

We introduce version 2 of the International System(s) Dataset (ISD), a register of sovereign states across the 1816–2016 period that include numerous states that are missed in commonly used datasets like the Correlates of War (COW) Project. Whereas ISD version 1 identified 363 states between 1816 and 2011, version 2 identifies 482. This version also records valuable information on a range of corollary variables, including start dates, end dates, estimated population sizes, diplomatic relations with Europe, conflict episodes, the existence of borders, and the location of capital cities. This dataset makes an important contribution to the study of international relations. It provides a more accurate understanding of the development of the international system over the last two centuries, it moves beyond the Eurocentric bias that sits at the heart of existing quantitative IR scholarship, and it will enable scholars to pursue a range of research topics such as the historical importance of state borders and boundaries, the practices surrounding recognition, and the frequency and intensity of conflict across regions. In this article, we discuss the existing state system membership lists and show how the ISD addresses their shortcomings. We outline the key concept and operationalization of statehood that the ISD adopts. We detail the variables included in this version of the ISD, discuss the data collection process, and show temporal and spatial distributions that illustrate the uniqueness of the ISD. Finally, we demonstrate the utility of bringing the ISD into one of many potential research topics: the study of conflict.     

Learning how to be efficient property owners and accountable governments: the case of Kyrgyzstan's cities

As a result of government decentralization and property devolution from central to local governments, many city governments in transition countries became the largest real property owners in urban areas. For transitional countries lacking sufficient democratic traditions, efficient asset management represents a dramatic and multifaceted challenge. Kyrgyzstan happened to be among the first transitional countries where cities obtained systematic technical assistance in adapting the most advanced methodology of municipal asset management, which was recently developed under the auspices of the World Bank. Based on experience of work within five pilot cities, the article summarizes factors and methods that help develop a sense of strong ownership and internalization of new asset management vision and approaches among local officials and elected members of city councils. The article presents the experience of how the government of Uzgen, a city of about 40,000 people in South Kyrgyzstan, has changed its attitudes and approach regarding management of municipal property. The article demonstrates how two key processes—improving professional aspects of municipal asset management and developing local democracy though public involvement—have been reinforced by each other. One of the main lessons from Uzgen's experience is that municipal property asset management—because it deals with tangible issues—is a perfect focal point for developing local democracy and communication between a local population and its government. Copyright © 2001 John Wiley & Sons, Ltd.     

Strategic Defiance and Compliance in the U.S. Courts of Appeals

Why do lower courts treat Supreme Court precedents favorably or unfavorably? To address this question, we formulate a theoretical framework based on current principal‐agent models of the judiciary. We use the framework to structure an empirical analysis of a random sample of 500 Supreme Court cases, yielding over 10,000 subsequent treatments in the U.S. Courts of Appeals. When the contemporary Supreme Court is ideologically estranged from the enacting Supreme Court, lower courts treat precedent much more harshly. Controlling for the ideological distance between the enacting and contemporary Supreme Courts, the preferences of the contemporary lower court itself are unrelated to its behavior. Hence, hierarchical control appears strong and effective. At the same time, however, a lower court's previous treatments of precedent strongly influence its later treatments. The results have important implications for understanding legal change and suggest new directions for judicial principal‐agency theory.     

Integrating ethics into the public administration curriculum: A three-step process

This article provides a three-step process for analyzing public policy dilemmas with ethical implications. A framework is proposed that builds on existing ethics theories and attempts to provide a relevant, usable approach for decisionmaking. A review of current thought in ethics indicates a concern for two areas: (a) responsibilities to relevant constituencies; and (b) adherence to moral obligations. The framework presented herein directly addresses both of these areas of concern. The authors have found this approach to be useful for classroom applications. This process is simple to explain, understand, and apply to a range of administrative situations. Students find the framework a memorable tool, useful in structuring deliberations with ethical implications. Sample applications of the framework provide examples for educators interested in integrating ethics into their advanced undergraduate and graduate courses.     

Policy Diffusion: The Issue‐Definition Stage

We put forward a new approach to studying issue definition within the context of policy diffusion. Most studies of policy diffusion—which is the process by which policymaking in one government affects policymaking in other governments—have focused on policy adoptions. We shift the focus to an important but neglected aspect of this process: the issue‐definition stage. We use topic models to estimate how policies are framed during this stage and how these frames are predicted by prior policy adoptions. Focusing on smoking restriction in U.S. states, our analysis draws upon an original data set of over 52,000 paragraphs from newspapers covering 49 states between 1996 and 2013. We find that frames regarding the policy's concrete implications are predicted by prior adoptions in other states, whereas frames regarding its normative justifications are not. Our approach and findings open the way for a new perspective to studying policy diffusion in many different areas.     

Blood factors of Sus scrofa following a series of three TASER electronic control device exposures

In a previous study, 18 repeated exposures of anaesthetized swine to an electro-muscular incapacitating device (TASER International's ADVANCED TASER X26 electronic control device) resulted in acidosis and increases in blood electrolytes. In the current study, experiments were performed to investigate effects of a more typical scenario of repeated exposures of the device on muscle contraction and changes in blood factors. Ten swine were exposed for 5s, followed by a 5-s period of no exposure, three times. Selected blood factors were monitored for 3h following exposure. Transient increases in blood glucose, lactate, sodium, potassium, calcium, and pCO(2) were consistent with previous reports in the literature dealing with studies of muscle stimulation or exercise. Blood pH was decreased immediately following exposure, but subsequently returned toward a normal level. Oxygen saturation (measured by pulse oximetry) was not changed significantly. In conclusion, three repeated TASER device exposures had only transient effects on blood factors, which all returned to pre-exposure levels, with the exception of hematocrit (which remained elevated after 3h). Since the increase in this factor was less than that which may occur after short periods of exercise, it is unlikely that this would be an indicator of any serious harm.     

Social Integration, Self-control, and Conformity

Evidence relevant to claims of self-control theory concerning the connection between social integration and crime/deviance is offered. Using data from a survey of the population of a southwestern city that permit measurement of two types of social integration, including socially supportive networks, we (1) investigate the association between self-control and social integration, and (2) attempt to ascertain if social integration is associated with misbehavior independently of self-control. Results suggest that self-control is a persistent predictor of misconduct that operates independently of social integration. Although interpersonal social integration appears to stand alone in its association with deviance, community integration shows no relationship with self-control or misbehavior.