Non-invasive fetal genotyping for maternal alleles with droplet digital PCR: A comparative study of analytical approaches

Objectives

To develop a flexible droplet digital PCR (ddPCR) workflow to perform non-invasive prenatal diagnosis via relative mutation dosage (RMD) for maternal pathogenic variants with a range of inheritance patterns, and to compare the accuracy of multiple analytical approaches.

Methods

Cell free DNA (cfDNA) was tested from 124 archived maternal plasma samples: 88 cases for sickle cell disease and 36 for rare Mendelian conditions. Three analytical methods were compared: sequential probability ratio testing (SPRT), Bayesian and z-score analyses.

Results

The SPRT, Bayesian and z-score analyses performed similarly well with correct prediction rates of 96%, 97% and 98%, respectively. However, there were high rates of inconclusive results for each cohort, particularly for z-score analysis which was 31% overall. Two samples were incorrectly classified by all three analytical methods; a false negative result predicted for a fetus affected with sickle cell disease and a false positive result predicting the presence of an X-linked IDS variant in an unaffected fetus.

Conclusions

ddPCR can be applied to RMD for diverse conditions and inheritance patterns, but all methods carry a small risk of erroneous results. Further evaluation is required both to reduce the rate of inconclusive results and explore discordant results in more detail.     

Ethical issues associated with prenatal screening using non-invasive prenatal testing for sex chromosome aneuploidy

Prenatal screening for sex chromosome aneuploidies (SCAs) is increasingly available through expanded non-invasive prenatal testing (NIPT). NIPT for SCAs raises complex ethical issues for clinical providers, prospective parents and future children. This paper discusses the ethical issues that arise around NIPT for SCAs and current guidelines and protocols for management. The first section outlines current practice and the limitations of NIPT for SCAs. It then outlines key guidelines before discussing the ethical issues raised by this use of NIPT. We conclude that while screening for SCAs should be made available for people seeking to use NIPT, its implementation requires careful consideration of what, when and how information is provided to users.     

Building institutions based on information disclosure: lessons from GRI's sustainability reporting

Global Reporting Initiative (GRI) is the best-known framework for voluntary reporting of environmental and social performance by business worldwide. Using extensive empirical data, including interviews and documentary analysis, we examine GRI's organizational field and conclude that since its modest beginnings in 1999 GRI has been by several measures a successful institutionalization project. But the institutional logic of this new entity, as an instrument for corporate sustainability management, leaves out one of the central elements of the initial vision for GRI: as a mobilizing agent for many societal actors. This emergent logic reflects GRI's dominant constituency – large global companies and financial institutions and international business management consultancies – and not the less active civil society organizations and organized labor. We attribute these developments to factors such as building GRI within the existing institutional structures; the highly inclusive multistakeholder process; and the underdeveloped base of information users. From the institutional theory perspective, this case shows how the process of institutionalization is deeply affected by initial strategies of the founders, and how it reproduces existing power relations. From the governance perspective, this case leads us to question the power of commodified information to mobilize civil society and to strengthen governance based on partnerships.     

Decision‐Making Triggers in Adaptive Management

We analyzed whether decision‐making triggers increase accountability of adaptive‐management plans. Triggers are prenegotiated commitments in an adaptive‐management plan that specify what actions are to be taken and when on the basis of information obtained from monitoring. Triggers improve certainty that particular actions will be taken by agencies in the future. We conducted an in‐depth, qualitative review of the political and legal contexts of adaptive management and its application by U.S. federal agencies. Agencies must satisfy the judiciary that adaptive‐management plans meet substantive legal standards and comply with the U.S. National Environmental Policy Act. We examined 3 cases in which triggers were used in adaptive‐management plans: salmon (Oncorhynchus spp.) in the Columbia River, oil and gas development by the Bureau of Land Management, and a habitat conservation plan under the U.S. Endangered Species Act. In all the cases, key aspects of adaptive management, including controls and preidentified feedback loops, were not incorporated in the plans. Monitoring and triggered mitigation actions were limited in their enforceability, which was contingent on several factors, including which laws applied in each case and the degree of specificity in how triggers were written into plans. Other controversial aspects of these plans revolved around who designed, conducted, interpreted, and funded monitoring programs. Additional contentious issues were the level of precaution associated with trigger mechanisms and the definition of ecological baselines used as points of comparison. Despite these challenges, triggers can be used to increase accountability, by predefining points at which an adaptive management plan will be revisited and reevaluated, and thus improve the application of adaptive management in its complicated political and legal context. Detonadores de la Toma de Decisiones en el Manejo Adaptativo     

Higher removal rate of eggs laid by anarchistic queens—a cost of anarchy?

Anarchy, where honeybee workers produce males in the presence of a queen, is extremely rare in natural honeybee populations, suggesting that there are colony-level costs associated with being anarchistic. Yet, no significant costs have yet been identified. A possible reason for this may be that researchers have only focused on the behaviour of anarchistic workers, which have been shown to perform worker-tasks as do wild-type workers. Possibly, therefore, costs associated with anarchy should be sought in anarchistic queens and not workers. A potential cost could be a lower survival rate of eggs laid by anarchistic queens perhaps because their egg-marking signal is not as clear as that of a wild-type queen. In this paper, we determined the removal rate of eggs laid by anarchistic queens in standard worker-policing bioassays. Our results show that eggs laid by anarchistic queens are removed at a higher rate than eggs laid by wild-type queens. This does not seem to be due to differences in hydrocarbons found on the surface of eggs, as both egg types showed the same alkanes and alkenes in similar proportions. We postulate that higher removal rates of queen-laid eggs due to recognition errors may be one reason that anarchy is rare in natural honeybee populations.