Increased serum concentrations of TIMP-4 and Endoglin and decreased levels of pentraxin-3 in CF patients with liver disease.

<p>In the whole CF cohort, patients with CFLD (n = 17) exhibited significantly higher serum levels of TIMP-4 and Endoglin than patients without liver disease (n = 28). Pentraxin-3 serum levels were significantly decreased in patients with CFLD whereas serum concentration of hepatocyte growth factor (HGF) was unchanged between CF patients with and without CFLD.</p

Increased serum levels of Endoglin and decreased concentration of pentraxin-3 in CF patients with increased liver stiffness. CF patients with liver stiffness values above a threshold value of 6.3 kPa, indicative for CFLD, exhibited significantly higher serum levels of Endoglin and significantly lower serum levels of pentraxin-3 compared to CF patients with liver stiffness values below 6.3 kPa.

<p>TIMP-4 was also increased in patients with a liver stiffness above the threshold of 6.3 kPa, although these elevations did not reach statistical significance. The cut-off value of 6.3 kPa for the identification of patients with CFLD was derived from ROC analyses as value at which the sum of diagnostic sensitivity and specificity for the detection of CFLD was maximal (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058955#pone-0058955-t003" target="_blank">Table 3</a>).</p

Demographic and clinical data of the CF patient cohort.

*<p>significantly different, P <0.05</p>**<p>significantly different, P <0.01</p><p>UDCA: Ursodeoxycholic Acid</p

<b>Table 3.</b> Diagnostic performances of novel diagnostic markers and clinical markers for the detection of CFLD.

#<p>age and gender specific cut-off, values determined by the Department for Laboratory Medicine and Clinical Chemistry of the University Hospital Giessen according to the International Federation of Clinical Chemistry</p><p>PPV: positive predictive value; NPV: negative predictive value; CI: confidence interval</p

Assessment of chemokines in patients with CFLD.

<p>Relative expression of 31 different chemokines regulating the migration of monocytes, neutrophils, and lymphocytes (A) were determined from pooled serum from patients with established CFLD (n = 4) and those without liver disease (n = 4). All proteins were determined in duplicate and based on optical densitometry of the corresponding bands (B), proteins were identified that were at least 2-fold differentially regulated in patients with CFLD compared to those without. CX3CL1, CXCL1, CCL1, IL-16, CXCL10 and MIP-3β were at least 2-fold increased in CFLD whereas IL-8 and CCL17 was at least 2-fold decreased in patients with CFLD compared to those without (C). ). <i>PC: positive control, NC: negative control, IC: internal control, OD: optical density</i></p

Assessment of soluble receptors and related proteins from non-hematopoietic cells in patients with CFLD (Array A).

<p>Relative expression of 62 different soluble receptors and related proteins from non-hematopoietic proteins (A) were determined from pooled sera from patients with established CFLD (n = 4) and those without liver disease (n = 4). All proteins were determined in duplicate and based on optical densitometry of the corresponding bands (B), proteins were identified that were at least 2-fold differentially regulated in patients with CFLD compared to those without. A total of 10 soluble receptors and related proteins from non-hematopoietic cells were at least 2-fold increased in CFLD with highest relative expression observed for MEPE and SREC-II (C). ITGA3, JAM-B, MUCDHL and SDC4 were at least 2-fold decreased in patients with CFLD compared to those without (C). <i>PC: positive control, NC: negative control, IC: internal control, OD: optical density</i></p

Concordance between clinical markers and novel diagnostic markers for CFLD.

<p>Measurement agreement between AST, ALT, γGT and ALP as commonly used clinical markers of CFLD and TIMP-4, Endoglin and transient elastography as novel diagnostic modalities was assessed in Bland-Altman Analyses. TIMP-4 and Endoglin showed very good agreement with ALT (A) and AST (B) levels. Comparison of the concordance of TE and levels of ALT (A), AST (B) and γGT (C) showed good agreement of the different methods although a greater bias was observed with higher average values. A proportional error was observed in the agreement of γGT and the biomarkers TIMP-4 and Endoglin (C) and in the agreement of ALP and TE (D). The dashed line shows 95% limits of agreement, the solid line indicates the systemic error (bias).</p

Diagnostic accuracy of novel diagnostic markers and clinical markers for the detection of CFLD.

<p>ROC: receiver operating characteristic; AUC: area under the curve; CI: confidence interval; APRI: AST/Platelets-Ratio-Index</p

Increased serum levels of TIMP-4 and Endoglin in HCV patients with complete liver cirrhosis.

<p>Serum levels of TIMP-4, Endoglin and Pentraxin-3 were comparable and without significant differences between patients with CFLD and those with HCV liver disease (top row, A). In the cohort of patients with HCV liver disease, patients with complete fibrosis/cirrhosis (F = 4) exhibited significantly increased serum levels of TIMP-4 and Endoglin compared those HCV patients with portal fibrosis with few or numerous fibrotic septae but without cirrhosis (F = 2/3), whereas Pentraxin-3 remained unchanged in the different HCV fibrosis stages (bottom row, B).</p

Baseline Characteristics^*.

<p>*presented as median values.</p><p>**nucleosid/nucleotide analogs.</p><p>***not determined.</p