Workshop report on the extraction of foetal DNA from maternal plasma

Objective Cell free foetal DNA (cff DNA) extracted from maternal plasma is now recognized as a potential source for prenatal diagnosis but the methodology is currently not well standardized. To evaluate different manual and automated DNA extraction methods with a view to developing standards, an International Workshop was performed. Methods Three plasma pools from RhD-negative pregnant women, a DNA standard, real-time-PCR protocol, primers and probes for RHD were sent to 12 laboratories and also to one company (Qiagen, Hilden, Germany). In pre-tests, pool 3 showed a low cff DNA concentration, pool 1 showed a higher concentration and pool 2 an intermediate concentration. Results The QIAamp DSP Virus Kit, the High Pure PCR Template Preparation Kit, an in-house protocol using the QIAamp DNA Blood Mini Kit, the CST genomic DNA purification kit, the Magna Pure LC, the MDx, the M48, the EZ1 and an in-house protocol using magnetic beads for manual and automated extraction were the methods that were able to reliably detect foetal RHD. The best results were obtained with the QIAamp DSP Virus Kit. The QIAamp DNA Blood Mini Kit showed very comparable results in laboratories that followed the manufacturer's protocol and started with ≥ 500 µL plasma. One participant using the QIAamp DNA Blood Midi Kit failed to detect reliably RHD in pool 3. Conclusions This workshop initiated a standardization process for extraction of cff DNA in maternal plasma. The highest yield was obtained by the QIAamp DSP Virus Kit, a result that will be evaluated in more detail in future studies. Copyright © 2007 John Wiley & Sons, Ltd.     

Sharing developed countries’ post-2012 greenhouse gas emission reductions based on comparable efforts

The Bali Action Plan as adopted under the United Nations Framework Convention on Climate Change (UNFCCC) in 2007, states that Annex I (developed) countries should reduce their greenhouse gas emissions, based on comparable efforts. Within this context, we have explored various comparable effort approaches (for example, equal marginal abatement costs for all countries) for reducing emissions by the year 2020 for individual countries and regions. In all calculations, the total reduction for Annex I countries as a group is assumed to be 30% below 1990 levels. In the analysis, we compare the reduction targets as calculated from the different approaches with the emission reductions as pledged by these countries as part of the Copenhagen Accord, as drafted under the UNFCCC in 2009. Our analysis indicates that the different elements in these calculations may cause a diversity in outcomes and that, therefore, individual countries may favour certain elements over others. These elements include (a) the choice of the approach itself (the same approach may produce very different outcomes for countries with diverging national circumstances, such as Canada and Russia); (b) the reference year (such as 1990 or 2006 emissions, is very important for countries with an increase in emissions since 1990 (e.g. the United States, Canada) or for those that have lower emission levels (e.g. Russia, the Ukraine)); and (c) rules on land use (these are important for countries with large forest areas). It should be noted that the stringency of the individual countries’ reductions as pledged, differs substantially from the stringency of the reduction targets calculated from the effort-sharing approaches. The current pledges by both the European Union and the United States, are lower than the reductions that would be obtained in the effort-sharing approaches for a 30% overall reduction in Annex I countries.     

Environmental Modeling and Methods for Estimation of the Global Health Impacts of Air Pollution

Air pollution is increasingly recognized as a significant contributor to global health outcomes. A methodological framework for evaluating the global health-related outcomes of outdoor and indoor (household) air pollution is presented and validated for the year 2005. Ambient concentrations of PM2.5 are estimated with a combination of energy and atmospheric models, with detailed representation of urban and rural spatial exposures. Populations dependent on solid fuels are established with household survey data. Health impacts for outdoor and household air pollution are independently calculated using the fractions of disease that can be attributed to ambient air pollution exposure and solid fuel use. Estimated ambient pollution concentrations indicate that more than 80% of the population exceeds the WHO Air Quality Guidelines in 2005. In addition, 3.26?billion people were found to use solid fuel for cooking in three regions of Sub Saharan Africa, South Asia and Pacific Asia in 2005. Outdoor air pollution results in 2.7?million deaths or 23?million disability adjusted life years (DALYs) while household air pollution from solid fuel use and related indoor smoke results in 2.1?million deaths or 41.6?million DALYs. The higher morbidity from household air pollution can be attributed to children below the age of 5 in Sub Saharan Africa and South Asia. The burden of disease from air pollution is found to be significant, thus indicating the importance of policy interventions.