Selected box plots for metabolites of interest for PD (left) and RLS (right).

<p>Selected box plots for metabolites of interest for PD (left) and RLS (right).</p

Gaussian graphical model of serum amino acids and acylcarnitine metabolite concentrations in KORA S4.

<p>Each node represents a metabolite, whereas edges represent significant partial correlations. Nodes were coloured according to the β-estimate and the p-value from the linear models (red  =  positive association with FFMI; blue  =  negative association with FFMI; white  =  not significant association with FFMI).</p

Characteristics (mean ± SD; % or n absolute) of male and female subjects in the analysed samples of KORA S4 and F4.

a<p>Parameters derived from the bioelectrical impedance analysis measurements which were only conducted in KORA S4;</p>b<p>Fat Free Mass Index;</p>c<p>Body Fat Mass Index;</p>d<p>Sports (active: >1 h leisure time of sports per week on a regular basis; inactive: less than 1 h sports per week).</p

Boxplot of serum BCAA concentrations (µmol/l), by quintiles of the FFMI, for the KORA S4 population.

<p>Boxplot of serum BCAA concentrations (µmol/l), by quintiles of the FFMI, for the KORA S4 population.</p

Inverse normalized values of aspartylphenylalanine for KORA and TwinsUK (AA: major homozygotes, AG: heterozygotes, GG minor homozygotes).

<p>Inverse normalized values of aspartylphenylalanine for KORA and TwinsUK (AA: major homozygotes, AG: heterozygotes, GG minor homozygotes).</p

Selected metabolic traits significantly associated with FFMI^a in a linear regression model adjusted for age, and sex for the non-obese and obese participants in the KORA S4 sample.

a<p>Fat Free Mass Index;</p>b<p>for multiple testing adjusted p-value;</p>c<p>adjusted R² of the linear model; AAs, amino acids; Σ aromatic amino acids, sum of tyrosine, phenylalanine, and tryptophan; Σ BCAAs, sum of valine, isoleucine, and leucine; Σ glucogenic amino acids, sum of alanine, glycine, and serine.</p

Significant results from the KORA F4 analysis for rs4329.

<p>Significant results from the KORA F4 analysis for rs4329.</p

Summary of here discussed genotype dependent polypeptide cleavage catalyzed by ACE.

<p>Blue coloring: health effect of the respective polypeptide; green: measured metabolites.</p

Twenty-two identified and replicated loci and their overlap with associations to metabolic traits and clinical phenotypes.

<p>^a SNP with the strongest association to targeted metabolic traits (“lead SNP”)</p><p>^b SNP with the strongest association to non-targeted metabolic traits</p><p>^c manually added to the list of most plausible candidate genes derived by evidence-based selection</p><p>^d additional candidates match to other non-targeted traits that also associate with the lead SNP</p><p>^e results from GWAS (<i>P</i> < 5.0×10⁻⁸)</p><p>^f mutations determined in clinical studies</p><p>For each locus, we selected all variants that displayed genome-wide significant association signals to metabolic traits in the SHIP-0 cohort. We added their proxy variants in LD (r² ≥ 0.8; based on 1000 genomes project data [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.ref050" target="_blank">50</a>, <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.ref051" target="_blank">51</a>]). These variant sets were used for the selection of candidate genes and the comparison with association results from other studies. <b>Candidate genes:</b> selection of genes based on variant evidence (genes hit or close-by, eQTL, potentially regulatory effects, or missense variants) (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.s007" target="_blank">S3 Table</a>). Genes with the highest evidence counts are listed. Genes with the most plausible biochemical relation to the associated trait are highlighted in bold typeface. <b>Associated traits</b>: Targeted/non-targeted metabolomics: traits that display genome-wide significant association signals in SHIP-0. The arrows indicate whether the trait increases (↗) or decreases (↘) per copy of the effect allele. For non-targeted traits, the most plausible metabolite candidates according to metabomatching are given. <b>Other mGWAS in urine/blood:</b> metabolic traits that were previously found to be associated with a locus variant. The arrows indicate the directionality of the effect for the reported effect allele (where available). <b>Clinical phenotypes:</b> overlap with variants found to be associated with clinical traits. <b>Comment</b>: Gene expression rates were taken from the Illumina Body Map 2.0 (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.s008" target="_blank">S4 Table</a>). Protein localizations were taken from the Human Protein Atlas (version 12) [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.ref052" target="_blank">52</a>]. For genes linked to clinical traits, we provide OMIM or OrphaNet accession numbers if available. <b>Functional match</b>: Indicates which associations exhibit a sound biological link between gene function and the biochemical nature of the associated metabolite(s).</p><p>Twenty-two identified and replicated loci and their overlap with associations to metabolic traits and clinical phenotypes.</p

Fifteen genetic loci as discovered in the SHIP-0 data set and their most significant associations to targeted metabolic traits.

<p><b>Chr/Position</b>: Chromosomal location of the SNP according to the human reference genome (GRCh37). <b>EA/EAF</b>: Effect allele and frequency. <b>Trait or pairwise ratio</b>: Tested metabolic trait. In case of ratios, the trait that shows the stronger association signal is in the numerator. <b>N</b>: Number of samples for which both genotype and phenotype data were available for the tested SNP/metabolic trait pair. <b>beta’</b>: beta’ is defined as 10^beta-1 where beta depicts the relative effect size representing the slope of the regression line in the linear model when using log₁₀-scaled metabolic traits and the occurrence of the SNP’s minor allele (coded as 0,1, and 2). Thus, beta’ describes the relative difference per minor allele copy for non-scaled metabolic traits in comparison to the estimated mean of the metabolic trait in the major homozygote test subjects. <b><i>P</i>-gain</b>: Defined as min(<i>P</i>(M₁)/<i>P</i>(M₁/M₂), <i>P</i>(M₂)/<i>P</i>(M₁/M₂)), where M₁ and M₂ represent the two traits of which the ratio M₁/M₂ is built. <b>Replicated</b>: SNP/metabolic trait pair was replicated in KORA F4 (<i>P <</i> 1.32×10⁻³) (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.s005" target="_blank">S1 Table</a>). <b>Non-targeted</b>: SNP or proxy in linkage disequilibrium (LD) is also associated with a non-targeted metabolic trait (<i>P <</i> 3.25×10⁻¹²) (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1005487#pgen.1005487.t002" target="_blank">Table 2</a>).</p><p>Fifteen genetic loci as discovered in the SHIP-0 data set and their most significant associations to targeted metabolic traits.</p