Serum markers detect the presence of liver fibrosis: A cohort study William M.C. Rosenberg, Michael Voelker, Robert Thiel, Michael Becka, Alastair Burt, Detlef Schuppan, Stefan Hubscher, Tania Roskams, Massimo Pinzani, Michael J.P. Arthur  Gastroenterology  Volume 127, Issue 6, Pages 1704-1713 (December 2004) DOI: 10.1053/j.gastro.2004.08.052 Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 1 Flow chart depicting the recruitment and participation of subjects in the European Liver Fibrosis study. Subjects were recruited consecutively at all centers. All biopsy specimens that met inclusion criteria were staged by central pathologist A. Pathologists B and C each staged a subset of 620 biopsy specimens selected at random from the group staged by pathologist A, generating 4 sets of scores: RA1, RA2, RB, and RC. The whole cohort (GA) was divided into the test (GT) and validation (GV) sets for derivation of the algorithm and subsequent validation. Gastroenterology 2004 127, 1704-1713DOI: (10.1053/j.gastro.2004.08.052) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 2 (A) Box-and-whisker plot for GV; Scheuer fibrosis score versus algorithm discriminant score. The middle horizontal line in each box is the median, and the notched region denotes the estimated 95% CI on the median. The end lines are the 25th and 75th percentiles. The dashed lines indicate the acceptable range for data. Crosses and circles indicate potential outliers. The range of algorithm scores was from −3.5 to +6.0. The principal discriminant function for each staging system was as follows: Scheuer D = −.014·LN(age) + .616·LN(HA) + .586·LN(PIIINP) + .472·LN(TIMP-1) − 6.38; Ishak D = −.08·LN(age) + .608·LN(HA) + .601·LN(PIIINP) + .511·LN(TIMP-1) − 6.26. (B–E) Box-and-whisker plot for GV; Scheuer fibrosis score versus natural logarithm of individual component marker scores of the algorithm. The middle horizontal line in each box is the median, and the notched region denotes the estimated 95% CI on the median. The end lines are the 25th and 75th percentiles. The dashed lines indicate the acceptable range for data. Crosses indicate potential outliers. B shows data for age, C for TIMP-1, D for PIIINP, and E for HA. Gastroenterology 2004 127, 1704-1713DOI: (10.1053/j.gastro.2004.08.052) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 3 Cumulative percent distributions of the principal discriminant function scores by Scheuer stage. For example, for a Scheuer stage of 0, approximately 65% of all patients in this group had a discriminant score of −.5 or less. The Euclidian distance between score groups was calculated for the separation of scores that corresponded to each stage. This analysis showed the natural bifurcations reflected in the graphs plotted above. To calculate the distances between score groups, the group centroid is defined as the value of the discriminant function when the means of all variables are used in the equation. So, in Scheuer stage 0, the group centroid would be D0 = .014·LN(mean[age]) + .616·LN(mean[HA]) + .586·LN(mean[PIIINP]) + .472·LN(mean[TIMP-1]) − 6.38. The Euclidian distance between 2 groups is defined as Delta(i, j) = Di2+Dj2. Gastroenterology 2004 127, 1704-1713DOI: (10.1053/j.gastro.2004.08.052) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 4 Box-and-whisker plot for GV showing Scheuer fibrosis score versus discriminant score. The middle horizontal line is the median, and the notched region denotes the estimated 95% CI on the median. The end lines are the 25th and 75th percentiles. The dashed lines indicate the acceptable range for data. Crosses indicate potential outliers. The data plotted show the distribution of discriminant scores for biopsy specimens of Scheuer stage 0–2 and 3–4. Gastroenterology 2004 127, 1704-1713DOI: (10.1053/j.gastro.2004.08.052) Copyright © 2004 American Gastroenterological Association Terms and Conditions

Figure 5 ROC curve for GV. The ROC curve is shown for the performance of the algorithm combining serum markers for discriminating between Scheuer fibrosis stages 0–2 and 3–4 in the GV sample set assessed by pathologist A. Gastroenterology 2004 127, 1704-1713DOI: (10.1053/j.gastro.2004.08.052) Copyright © 2004 American Gastroenterological Association Terms and Conditions