Whitfield Lynge (archermonday2)

Life is filled with puzzles and mysteries, and we often fail to recognize the difference. As described by Gregory Treverton and Malcolm Gladwell, puzzles are solved by gathering and assimilating all relevant data in a logical, linear fashion, as in deciding which antibiotic to prescribe for an infection. check details In contrast, mysteries remain unsolved until all relevant data are analyzed and interpreted in a way that appreciates their depth and complexity, as in determining how to best modulate the host immune response to infection. When investigating mysteries, we often fail to appreciate their depth and complexity. Instead, we gather and assimilate more data, treating the mystery like a puzzle. This strategy is often unsuccessful. Traditional approaches to predictive analytics and phenotyping in surgery use this strategy.Since humans have two copies of each gene, multiple mutations in different loci may or may not be found on the same strand of DNA (i.e., inherited from one parent). When a person is heterozygous at more than one position, the placement of these mutations, also called the haplotype phase, (i.e., cis for the same strand and trans for different strands) can result in the expression of different amount and type of proteins. In this work, we described an enzyme-free method to phase two single nucleotide polymorphisms (SNPs) using two fluorophore/quencher-labelled probes, where one of which was biotinylated. The fluorescence signal was obtained twice first, after the addition of the labelled probes and second, after the addition of the magnetic beads. The first signal was shown to be proportional to the total number of SNP A and SNP B present in the target analyte, while the second signal showed a marked decrease of the fluorescence signal from the non-biotinylated probe when the SNPs were in trans, showing that the probe immobilized on the magnetic bead selectively captures targets with SNPs in a cis configuration. We then mimic the nature of the human genome which consists of two haplotype copies of each gene, and showed that 250 nM of the 10 possible pairs of haplotypes could be differentiated using a combination of fluorescence microscopy and fluorescence detection.Insufficient method repeatability is a problem characterising the evaluation of certified reference materials (CRMs). In investigating the homogeneity studies of 216 certified parameters from 36 CRMs released by the European Commission's Joint Research Centre (JRC) over the last four years, it was found that in 1/3 of the cases, the method repeatability (sr) was too high to calculate the standard deviation between units (sbb) by classical analysis of variance (ANOVA). It was also found that the application of the repeatability requirement stated in the ISO Guide 352017 is not feasible since it would require unrealistically low repeatability standard deviations or an impossibly high number of replicates per unit. Evaluation of the uncertainty of homogeneity (ubb) as evaluated by ANOVA using both the maximum of sbb and 0, the maximum of sbb and u∗bb, the uncertainty hidden by method repeatability, the maximum of sbb and sbb/√n and Bayesian analysis, using both informative and diffuse priors, as well as the staner- and underestimation of ubb is evaluated, and an appropriate approach is chosen based on this analysis.A fast and efficient selective pressurized liquid extraction (sPLE) method was developed to extract secondary metabolites from complex plant matrix. Convallaria majalis L., a plant producing toxic steroids, was used as proof-of-concept. The method was optimized in the aspects of preheating, dispersant, extraction temperature and solvent, and the use of C18 as in-cell cleanup sorbent. Eight authentic natural steroids with diverse sugar moieties and hydrophobicities were selected as reference analytes and spiked to 0.1 g dried leaves. The extraction performance was evaluated based on the analytes' stability, recovery, matrix effect in