Serum proteome profiling identifies novel and powerful markers of cystic fibrosis liver disease.

Cystic Fibrosis associated liver disease (CFLD) develops in approximately 30% of CF patients. However, routine sensitive diagnostic tools for CFLD are lacking. Within this study, we aimed to identify new experimental biomarkers for the detection of CFLD. 45 CF patients were included in the study and received transient elastography. Differential regulation of 220 different serum proteins was assessed in a subgroup of patients with and without CFLD. Most interesting candidate proteins were further quantified and validated by ELISA in the whole patient cohort. To assess a potential relation of biomarker expression to the degree of hepatic fibrosis, serum biomarkers were further determined in 18 HCV patients where liver histology was available. 43 serum proteins differed at least 2-fold in patients with CFLD compared to those without liver disease as identified in proteome profiling. In ELISA quantifications, TIMP-4 and Endoglin were significantly up-regulated in patients with CFLD as diagnosed by clinical guidelines or increased liver stiffness. Pentraxin-3 was significantly decreased in patients with CFLD. Serum TIMP-4 and Endoglin showed highest values in HCV patients with liver cirrhosis compared to those with fibrosis but without cirrhosis. At a cut-off value of 6.3 kPa, transient elastography compassed a very high diagnostic accuracy and specificity for the detection of CFLD. Among the biomarkers, TIMP-4 and Endoglin exhibited a high diagnostic accuracy for CFLD. Diagnostic sensitivities and negative predictive values were increased when elastography and TIMP-4 and Endoglin were combined for the detection of CFLD. Serum TIMP-4 and Endoglin are increased in CFLD and their expression correlates with hepatic staging. Determination of TIMP-4 and Endoglin together with transient elastography can increase the sensitivity for the non-invasive diagnosis of CFLD

Interleukin 6, lipopolysaccharide-binding protein and interleukin 10 in the prediction of risk and etiologic patterns in patients with community-acquired pneumonia: results from the German competence network CAPNETZ

<p>Abstract</p> <p>Background</p> <p>The aim of our study was to investigate the predictive value of the biomarkers interleukin 6 (IL-6), interleukin 10 (IL-10) and lipopolysaccharide-binding protein (LBP) compared with clinical CRB and CRB-65 severity scores in patients with community-acquired pneumonia (CAP).</p> <p>Methods</p> <p>Samples and data were obtained from patients enrolled into the German CAPNETZ study group. Samples (blood, sputum and urine) were collected within 24 h of first presentation and inclusion in the CAPNETZ study, and CRB and CRB-65 scores were determined for all patients at the time of enrollment. The combined end point representative of a severe course of CAP was defined as mechanical ventilation, intensive care unit treatment and/or death within 30 days. Overall, a total of 1,000 patients were enrolled in the study. A severe course of CAP was observed in 105 (10.5%) patients.</p> <p>Results</p> <p>The highest IL-6, IL-10 and LBP concentrations were found in patients with CRB-65 scores of 3-4 or CRB scores of 2-3. IL-6 and LBP levels on enrollment in the study were significantly higher for patients with a severe course of CAP than for those who did not have severe CAP. In receiver operating characteristic analyses, the area under the curve values for of IL-6 (0.689), IL-10 (0.665) and LPB (0.624) in a severe course of CAP were lower than that of CRB-65 (0.764) and similar to that of CRB (0.69). The accuracy of both CRB and CRB-65 was increased significantly by including IL-6 measurements. In addition, higher cytokine concentrations were found in patients with typical bacterial infections compared with patients with atypical or viral infections and those with infection of unknown etiology. LBP showed the highest discriminatory power with respect to the etiology of infection.</p> <p>Conclusions</p> <p>IL-6, IL-10 and LBP concentrations were increased in patients with a CRB-65 score of 3-4 and a severe course of CAP. The concentrations of IL-6 and IL-10 reflected the severity of disease in patients with CAP. The predictive power of IL-6, IL-10 and LBP for a severe course of pneumonia was lower than that of CRB-65. Typical bacterial pathogens induced the highest LBP, IL-6 and IL-10 concentrations.</p

Hif-1α and Hif-2α synergize to suppress AML development but are dispensable for disease maintenance

Leukemogenesis occurs under hypoxic conditions within the bone marrow (BM). Knockdown of key mediators of cellular responses to hypoxia with shRNA, namely hypoxia-inducible factor-1α (HIF-1α) or HIF-2α, in human acute myeloid leukemia (AML) samples results in their apoptosis and inability to engraft, implicating HIF-1α or HIF-2α as therapeutic targets. However, genetic deletion of Hif-1α has no effect on mouse AML maintenance and may accelerate disease development. Here, we report the impact of conditional genetic deletion of Hif-2α or both Hif-1α and Hif-2α at different stages of leukemogenesis in mice. Deletion of Hif-2α accelerates development of leukemic stem cells (LSCs) and shortens AML latency initiated by Mll-AF9 and its downstream effectors Meis1 and Hoxa9. Notably, the accelerated initiation of AML caused by Hif-2α deletion is further potentiated by Hif-1α codeletion. However, established LSCs lacking Hif-2α or both Hif-1α and Hif-2α propagate AML with the same latency as wild-type LSCs. Furthermore, pharmacological inhibition of the HIF pathway or HIF-2α knockout using the lentiviral CRISPR-Cas9 system in human established leukemic cells with MLL-AF9 translocation have no impact on their functions. We therefore conclude that although Hif-1α and Hif-2α synergize to suppress the development of AML, they are not required for LSC maintenanc

Are roundabouts really safer than intersections?

The meeting place of roadways from different directions has a higher safety risk because of the large number of conflict points associated with it. Consequently, intersection crashes, in general, are more severe when compared to crashes reported at other places. The current trend in many developed countries is to replace regular intersections with roundabouts as a safety modification. However, in Switzerland only one study has been conducted in 1994 examining the safety of such roundabouts. We are currently unaware weather roundabouts increase safety or result in a greater confusion among drivers and other road users. To address this issue, this study focuses on comparing the safety at intersections and roundabouts in the canton of Zurich

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

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