The impact of absolute importance on prospective memory: Instructions matter

Prospective memory (ProM) is the ability to remember and carry out a planned intention in the future. ProM performance can be improved by instructing participants to prioritize the ProM task over the ongoing task. However, the improvement of ProM performance by emphasizing the relative importance typically restricted to situations in which the overlap between processing requirements of the ProM task and the ongoing task is low. Thus, additional processing resources are allocated to the ProM task and consequently, a cost emerges for the ongoing task. The aim of the present study was to further investigate this relationship. Participants were asked to respond to either semantic or perceptual ProM cues, which were embedded in a complex ongoing short term memory task. We manipulated absolute rather than relative importance by emphasizing the importance of the ProM task to half of the participants (i.e., without instructing them to prioritize it over the ongoing task). The results revealed that importance boosted ProM performance independent of the processing overlap between the ProM task and the ongoing task. Moreover, no additional cost was associated with absolute importance. These results challenge the view that importance always enhances the allocation of resources to the ProM task.

Next-generation sequencing of HIV-1 RNA genomes: determination of error rates and minimizing artificial recombination.

Next-generation sequencing (NGS) is a valuable tool for the detection and quantification of HIV-1 variants in vivo. However, these technologies require detailed characterization and control of artificially induced errors to be applicable for accurate haplotype reconstruction. To investigate the occurrence of substitutions, insertions, and deletions at the individual steps of RT-PCR and NGS, 454 pyrosequencing was performed on amplified and non-amplified HIV-1 genomes. Artificial recombination was explored by mixing five different HIV-1 clonal strains (5-virus-mix) and applying different RT-PCR conditions followed by 454 pyrosequencing. Error rates ranged from 0.04-0.66% and were similar in amplified and non-amplified samples. Discrepancies were observed between forward and reverse reads, indicating that most errors were introduced during the pyrosequencing step. Using the 5-virus-mix, non-optimized, standard RT-PCR conditions introduced artificial recombinants in a fraction of at least 30% of the reads that subsequently led to an underestimation of true haplotype frequencies. We minimized the fraction of recombinants down to 0.9-2.6% by optimized, artifact-reducing RT-PCR conditions. This approach enabled correct haplotype reconstruction and frequency estimations consistent with reference data obtained by single genome amplification. RT-PCR conditions are crucial for correct frequency estimation and analysis of haplotypes in heterogeneous virus populations. We developed an RT-PCR procedure to generate NGS data useful for reliable haplotype reconstruction and quantification.