IKK2-deficient hepatocytes are more sensitive to bile acid induced apoptosis compared to wild type cells.

<p>(A) Representative images of DAPI stained primary WT and IKK2-KO hepatocyte cultures. Cells were left untreated or were stimulated with taurolithocholic acid 3-sulfate disodium salt (TLCS, 100 µM) for the indicated time periods. Arrowheads show apoptotic hepatocytes as indicated by smaller highly fluorescent nuclei with condensed chromatin. Scale bars, 100 µm. (B) Graphs showing the average proportion of apoptotic hepatocytes from 3 independent experiments. Mean values ± SEM shown. * p<0,05; ** p<0,01; n.s.: not statistically significant. (C) Immunoblot detection of cleaved Caspase-3. Primary hepatocytes were stimulated with glycochenodeoxy-cholate (GCDC, 50 µM, top) or TLCS (100 µM, bottom) for the indicated time periods. Tubulin serves as loading control.</p

Mdr2^−/−IKK2^Hep-KO mice develop severe fibrosis.

<p>(A) Representative images of Masson trichrome stained livers from 12-week-old littermate male mice and from 27–30 week-old male mice. (B) Representative images of Sirius Red stained livers from 12-week-old littermate male mice and from old adult male mice (27–30 weeks). Insert: High magnification of corresponding liver sections. Arrowhead: Pericellular fibers. Double arrowhead: Pericellular space free of stained fibers. Genotypes as in (A). (C) Quantification of fibrosis in mice with the depicted genotypes measured as percentage of Sirius Red positive area as a fraction of the total area of at least 10 high power fields per mouse. Left: young adult mice (males, 8–19 weeks), right: old adult mice (males, 20–42 weeks). Error bars indicate SEM. Scale bars, 100 µm.</p

Mdr2^−/−IKK2^Hep-KO mice exhibit increased liver damage, hepatocyte death and cholestasis.

<p>(A) Alanine aminotransferase (ALT), (B) alkaline phosphatase (AP), and (C) bilirubin levels in serum of adult male mice (8–26 weeks) with the indicated genotypes. Error bars indicate SEM. Number of mice: Mdr2^−/−IKK2^Hep−KO: n = 10, Mdr2^−/−: n = 7, Mdr2^+/−IKK2^Hep-KO: n = 7 and Mdr2^+/+IKK2^FL: n = 2. (D–E) Representative pictures of TUNEL (D) and PCNA staining (E) on sections of paraffin-embedded livers from mice with the indicated genotypes. The green signal in E corresponds to background auto-fluorescence and was used to visualize the general morphology of the liver tissues. Scale bars: 20 µm.</p

Histological analysis of the liver pathology in Mdr2^−/−IKK2^Hep-KO mice.

<p>(A) Representative images of Hematoxylin & Eosin (HE) stained livers from young (12-week-old) littermate male mice and old (27–30 weeks) mice with the specified genotypes. Insert: High magnification of corresponding liver sections. (B) Immunofluorescence staining of cytokeratin 19 (CK19) in old male mice. DNA was stained with DAPI. The dashed line marks the distance from one portal field to the next, showing extreme abundance of CK19 positive cells in the Mdr2^−/− IKK2^Hep-KO as compared to Mdr2^−/− or Mdr2^+/−IKK2^Hep-KO mice. PV portal vein. CV central vein. BD bile duct. Scale bars, 100 µm.</p

Neurophysiological and MRI data 3

The files contain neurophysiological and MRI (T2 weighted MRI, Diffusion Tensor Imaging) data from 20 patients with primary biliary cholangitis and 20 controls

Neurophysiological and MRI data 4

The files contain neurophysiological and MRI (T2 weighted MRI, Diffusion Tensor Imaging) data from 20 patients with primary biliary cholangitis and 20 controls

Neurophysiological and MRI data 1

The files contain neurophysiological and MRI (T2 weighted MRI, Diffusion Tensor Imaging) data from 20 patients with primary biliary cholangitis and 20 controls

Patients with primary biliary cholangitis and fatigue present with depressive symptoms and selected cognitive deficits, but with normal attention performance and brain structure

<div><p>Background</p><p>In primary biliary cholangitis (PBC) fatigue is a major clinical challenge of unknown etiology. By demonstrating that fatigue in PBC is associated with an impaired cognitive performance, previous studies have pointed out the possibility of brain abnormalities underlying fatigue in PBC. Whether structural brain changes are present in PBC patients with fatigue, however, is unclear. To evaluate the role of structural brain abnormalities in PBC patients severely affected from fatigue we, therefore, performed a case-control cerebral magnetic resonance imaging (cMRI) study and correlated changes of white and grey brain matter with the cognitive and attention performance.</p><p>Methods</p><p>20 female patients with PBC and 20 female age-matched controls were examined in this study. The assessment of fatigue, psychological symptoms, cognitive and attention performance included clinical questionnaires, established cognition tests and a computerized test battery of attention performance. T1-weighted cMRI and diffusion tensor imaging (DTI) scans were acquired with a 3 Tesla scanner. Structural brain alterations were investigated with voxel-based morphometry (VBM) and DTI analyses. Results were correlated to the cognitive and attention performance.</p><p>Results</p><p>Compared to healthy controls, PBC patients had significantly higher levels of fatigue and associated psychological symptoms. Except for an impairment of verbal fluency, no cognitive or attention deficits were found in the PBC cohort. The VBM and DTI analyses revealed neither major structural brain abnormalities in the PBC cohort nor correlations with the cognitive and attention performance.</p><p>Conclusions</p><p>Despite the high burden of fatigue and selected cognitive deficits, the attention performance of PBC patients appears to be comparable to healthy people. As structural brain alterations do not seem to be present in PBC patients with fatigue, fatigue in PBC must be regarded as purely functional. Future studies should evaluate, whether functional brain changes underlie fatigue in PBC.</p></div

Computerized test battery of attention performance.

<p>20 patients with PBC and 20 age- and sex-matched controls underwent a computerized test battery of attention performance (TAP) in order of the figure appearance. No differences of reaction time were observed between both groups within all subtests (Wilcoxon matched pairs test). Alertness I without and with warning signal (WS) (A), divided attention auditive/visual (B), 30 minutes vigilance test first half/second half (C), alertness II without and with warning signal (WS) (D). PBC, primary biliary cholangitis.</p