Multivariate analysis using PCA of the parameters in tables 1 and 2 (with BMI and age excluded) using two significant principal components based on eigenvalue criteria (>2).

<p>A. Females with ALS overlapped with female controls in the measured parameters. (t <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113619#pone.0113619-Andersen1" target="_blank">[1]</a>, 34% of variation vs. t <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113619#pone.0113619-Desport1" target="_blank">[2]</a>, 20% of variation; R2 = 0.67; Q2 = .25). B) The male ALS group was shifted from the male controls in the measured parameters. (t <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113619#pone.0113619-Andersen1" target="_blank">[1]</a>, 36% of variation vs. t <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0113619#pone.0113619-Desport1" target="_blank">[2]</a>, 15% of variation; R2 = 0.51; Q2 = 0.22).</p

Correlation between HDL triglycerides and time of survival in female patients (A); HDL triglycerides and time of survival in female patients (B) after exclusion of one long-surviving patient (shown in A).

<p>Correlation between HDL triglycerides and time of survival in female patients (A); HDL triglycerides and time of survival in female patients (B) after exclusion of one long-surviving patient (shown in A).</p

Female subjects Average values ± standard deviation for parameters measured for female controls and ALS patients, fold change (ALS/controls), and p-values for females.

<p>Female subjects Average values ± standard deviation for parameters measured for female controls and ALS patients, fold change (ALS/controls), and p-values for females.</p

Variability in the measured parameters in one female patient with sporadic pseudo bulbar paresis, the samples were collected over 11 months with one time-point in duplicate.

<p>Variability in the measured parameters in one female patient with sporadic pseudo bulbar paresis, the samples were collected over 11 months with one time-point in duplicate.</p

OPLS model predicting survival from the time of sampling for diet-matched ALS cases (p = 0.003).

<p>A. Survival predicted by model plotted against time of survival from sampling. B. Long survival from sampling correlated with higher levels of LDL cholesterol, total cholesterol, coenzyme Q, and VLDL cholesterol.</p

Male subjects Average values ± standard deviation for parameters measured for male controls, ALS patients, fold change (ALS/controls), and p-values for males.

<p>Male subjects Average values ± standard deviation for parameters measured for male controls, ALS patients, fold change (ALS/controls), and p-values for males.</p

Correlation to survival Pearson's correlation for the parameters plotted against survival from time of sampling for all subjects and Spearman's correlation for the parameters plotted against survival from time of sampling for females and males.

<p>Correlation to survival Pearson's correlation for the parameters plotted against survival from time of sampling for all subjects and Spearman's correlation for the parameters plotted against survival from time of sampling for females and males.</p

Case breakdown of the model building and model testing data sets within MS cohort included.

<p>Abbreviations: PPMS, primary progressive MS; SPMS, secondary progressive MS; RRMS, relapsing-remitting multiple sclerosis.</p

Key features of the selected MS biomarkers.

<p>Abbreviations: MS, multiple sclerosis; CSF, cerebrospinal fluids; MMP9, matrix metalloproteinase 9; CXCL13, chemokine (C–X–C motif) ligand 13; OPN, osteopontin; NFL, neurofilament-light chain.</p

PCA broadly distinguishes between inflammatory and non-inflammatory conditions.

<p>PCA scores plot for the model building dataset (Set 1). Most non-inflammatory cases (blue) are associated with positive scores, located in the upper part of the plot. There is more variation in the distribution of the inflammatory cases (yellow) but many associate with negative scores (<b>A</b>). The PCA loadings plot shows that all the measured variables except age underlie the observed scores distribution seen in A (<b>B</b>). Scores of the test set (Set 2) were predicted using the model derived for Set 1. Non-inflammatory conditions, again, cluster mostly in the upper part of the plot (<b>C</b>). Extreme outliers (≥3 SD) in both scores plots belonged to cases of herpes encephalitis and neuroborreliosis.</p