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Abnormal vascular compliance in patients with primary biliary cirrhosis
Poster Sessions
182
this 85 kDa glycoprotein, purified it from bile, tested its in vitro CCPA and measured its concentrations in patients with and without gallstones. Methods: LDP was prepared from CABF by discontinuous gradient ultracentrifugation. Proteins were analysed by SDS-PAGE, blotting and immunochemical staining with anti-carcinoembryonic antigen (CEA) antibodies crossreacting with carcinoembryonic antigen-related adhesion molecule 1 (CEACAM1). The 85 kDa isoform of CEACAM1 was immunopurified from hepatic bile and its CCPA was tested on model bile. Bile concentrations of 5 CEACAM proteins were determined in patients with and without gallstones. Results: Five CEA cross-reacting protein bands of 200, 115, 85, 50 and 40 kDa were identified in human bile. The 50 kDa band was increased in bile from patients without stones; there was no difference in the content of the other bands between stone patients and non-stone patients. All bands were also present in CABF, but only a subfraction of the 85 kDa band was resistant to digestion by pronase. This subfraction was the pronase resistant protein in LDP. Conclusion: We have identified the protein present in LDP to be CEACAM1-85.
•7
QUANTITATIVE AND QUALITATIVE EVALUATION OF BILIARY ALPHA-1-ANTITRYPSIN
S.S.J. Janciauskiene, D. Ruta, Z. Inga, L. Stefan. Lund University,
Methods: The Alx of 10 female patients with PBC (mean age = 54.6 years) was recorded as a measure of the arterial stiffness following a 10 hour fast. 13 age matched healthy females served as controls (mean age = 53.5 years). Results: The A1x was significantly higher in the PBC group compared to controls (mean = 35.8%, range = 16%-47%; mean = 26.1%, range = 11%-33%: P = 0.015). Confounding factors including age (mean = 54.6 yr, range = 31 yr-70 yr; mean = 56.1 yr, range = 49 yr-61 yr: P = 0.696), pulse rate (mean = 72.3, range 59-87; mean = 71.3, range = 62-83: P = 0.76) and blood pressure controls (MAP: mean = 100.6, range = 69-129; mean = 98.2, range = 85-100: P = 0.71) showed no significant difference between the groups Conclusions: These results confirm using a non-invasive, in vivo technique that there is abnormal vascular stiffness in PBC. Endothelial dysfunction may have clinically significant effects on the peripheral vascular, but whether this vascular abnormality is of significance in the pathogenesis of liver disease is unknown.
~ - 7 - ] URSODEOXYCHOLIC ACID (UDCA) FEEDING AGGRAVATES LIVER INJURY IN BILE DUCT-LIGATED AND MDR2 KOCKOUT MICE P. Fickert, G. Zollner, A. Fuchsbichler, C. Stumptner, F. Lammert, K. Zaltloukal, H. Denk, M. Trauner. Internal Medicine, Karl-Franzens
Sweden
University, Austria
Alpha-l-antitrypsin (AAT) is a serine proteinase inhibitor, and its concentration in duodenal contents and in gall bladder is ranging from 50 to 400 mikrog/mL. According to cholesterol crystal growth assays in vitro, biliary AAT has no cholesterol crystallisation-promoting activity. The hydrophobic peptide(s) of AAT are also found in human bile, and in a bile model are known to promote cholesterol crystallisation. The aim of this study was to analyse the levels and molecular profile of AAT in gall bladder bile samples obtained during laparoscopic cholecystectomy. Bile samples were obtained from 23 patients with gallstones, 7 of which had cholestasis and 4 had both cholestasis and cholangitis; mean age 59.5 years. The concentration of AAT was estimated by immunoelectrophoresis and molecular forms were determined by 12.5% SDS-PAGE electrophoresis followed with Western Blot analysis. Blots were developed using antisera against AAT or C-terminal fragment of AAT (C-36). We found higher levels of AAT (3.6--0.4 g/L) in 17% and very low levels (0.0(04).08 g/L) in 83% of all samples. The C-36 peptide was found in 9 of 23 analysed samples, and 6 of those had very low concentrations of AAT. Moreover, the peptide was found in patients diagnosed for gallstones but not in subgroups with cholestasis, cholangitis or both. The levels of cleaved AAT correlate with low concentrations of native AAT in gall bladder bile and suggest that the generation of cleaved forms of AAT may affect gallstone formation in vivo.
Background & Aim: It is unclear whether UDCA improves liver injury caused by bifiary obstruction. Therefore, it was the aim to study the effects of UDCA in obstructive cholestasis in mice. Methods: Mice were fed a UDCA (0.5% w/w) supplemented diet or control diet for 7 d. Thereafter mice were subjected to common bile duct ligation (CBDL; n = 24) or selective bile duct ligation (SBDL; n = 12) and continued on UDCA or control diet for further 3 d. In addition, naive mice were subjected to UDCA after CBDL (n = 6. (Furthermore, Mdr2 - / - (with non-suppurative cholangitis) were fed UDCA (0.5% w/w) for 7 d. Mortality rates, serum liver enzymes, total serum bile acid levels, bile composition, and liver histology were investigated. Results: CBDL and SBDL (ligated lobes) resulted in disseminated necrosis, bile infarcts, and periductular fibrosis, whereas non-ligated lobes remained unchanged. CBDL (3 days) in UDCA-fed mice aggravated necrosis and significantly increased mortality rates (7/12 vs 1/12 in controls; p < 0.05). UDCA feeding to SBDL mice aggravated necrosis exclusively in ligated lobes, whereas non-ligated remained unchanged. UDCA feeding started after CBDL resulted in 100% mortality after 5 days (6/6). Moreover, UDCA feeding resulted in bile infarcts in Mdr2 - / - . UDCA feeding had no effect on liver histology and liver enzymes in sham operated animals and wild type mice. Summary & Conclusions: UDCA aggravates liver injury in obstructive cholestasis in mice. Further clinical studies will have to determine potential UDCA side effects in patients with obstructive cholestasis.
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ABNORMAL VASCULAR COMPLIANCE IN PATIENTS WITH PRIMARY SILIARY CIRRHOSIS
I.S.H. Cadden, B.M. Mullah, M.E. Callender, I,S. Young. Dept of Medicine, Royal Victoria Hospital, Belfast, Northern Ireland, UK Introduction: Elevated serum markers have suggested endothelial dysfunction in Primary Biliary Cirrhosis (PBC). We used the technique of Pulse Wave Analysis (PWA) to study the arterial stiffness (increased in endothelial dysfunction) in patients with PBC. PWA uses applanation tonometery to record the radial artery pressure waveform non-invasively. Application of a reverse transfer function allows accurate reconstruction of the central aortic waveform and determination of the central aortic pressures. Augmentation of this pressure thus provides a measurement of the compliance of the vascular tree, and this can be expressed quantitatively as the augmentation index (Alx).
~-]
HEPATIC UPTAKE OF CHOLECYSTOKININ OCTAPEPTIDE (CCK-8) BY ORGANIC ANION TRANSPORTING POLYPEPTIDES OATP4 AND OATP8 OF RAT AND HUMAN LIVER
M.G. Ismair, V. Cattori, B. Hagenbuch, P.J. Meier, G.A. Kullack-Ublick.
Department of Medicine, University Hospital, Switzerland Background & Aims: Cholecystokinin (CCK) is a major gastrointestinal peptide hormone which is released postprandially from the small intestine and which exerts marked effects on gallbladder and gastrointestinal motility. The smaller isoforms CCK-8 and CCK-4 are rapidly taken up into hepatocytes, metabolized and finally excreted into bile. Our aim was to identify and characterize the hepatocellular CCK-8 uptake system.
182
this 85 kDa glycoprotein, purified it from bile, tested its in vitro CCPA and measured its concentrations in patients with and without gallstones. Methods: LDP was prepared from CABF by discontinuous gradient ultracentrifugation. Proteins were analysed by SDS-PAGE, blotting and immunochemical staining with anti-carcinoembryonic antigen (CEA) antibodies crossreacting with carcinoembryonic antigen-related adhesion molecule 1 (CEACAM1). The 85 kDa isoform of CEACAM1 was immunopurified from hepatic bile and its CCPA was tested on model bile. Bile concentrations of 5 CEACAM proteins were determined in patients with and without gallstones. Results: Five CEA cross-reacting protein bands of 200, 115, 85, 50 and 40 kDa were identified in human bile. The 50 kDa band was increased in bile from patients without stones; there was no difference in the content of the other bands between stone patients and non-stone patients. All bands were also present in CABF, but only a subfraction of the 85 kDa band was resistant to digestion by pronase. This subfraction was the pronase resistant protein in LDP. Conclusion: We have identified the protein present in LDP to be CEACAM1-85.
•7
QUANTITATIVE AND QUALITATIVE EVALUATION OF BILIARY ALPHA-1-ANTITRYPSIN
S.S.J. Janciauskiene, D. Ruta, Z. Inga, L. Stefan. Lund University,
Methods: The Alx of 10 female patients with PBC (mean age = 54.6 years) was recorded as a measure of the arterial stiffness following a 10 hour fast. 13 age matched healthy females served as controls (mean age = 53.5 years). Results: The A1x was significantly higher in the PBC group compared to controls (mean = 35.8%, range = 16%-47%; mean = 26.1%, range = 11%-33%: P = 0.015). Confounding factors including age (mean = 54.6 yr, range = 31 yr-70 yr; mean = 56.1 yr, range = 49 yr-61 yr: P = 0.696), pulse rate (mean = 72.3, range 59-87; mean = 71.3, range = 62-83: P = 0.76) and blood pressure controls (MAP: mean = 100.6, range = 69-129; mean = 98.2, range = 85-100: P = 0.71) showed no significant difference between the groups Conclusions: These results confirm using a non-invasive, in vivo technique that there is abnormal vascular stiffness in PBC. Endothelial dysfunction may have clinically significant effects on the peripheral vascular, but whether this vascular abnormality is of significance in the pathogenesis of liver disease is unknown.
~ - 7 - ] URSODEOXYCHOLIC ACID (UDCA) FEEDING AGGRAVATES LIVER INJURY IN BILE DUCT-LIGATED AND MDR2 KOCKOUT MICE P. Fickert, G. Zollner, A. Fuchsbichler, C. Stumptner, F. Lammert, K. Zaltloukal, H. Denk, M. Trauner. Internal Medicine, Karl-Franzens
Sweden
University, Austria
Alpha-l-antitrypsin (AAT) is a serine proteinase inhibitor, and its concentration in duodenal contents and in gall bladder is ranging from 50 to 400 mikrog/mL. According to cholesterol crystal growth assays in vitro, biliary AAT has no cholesterol crystallisation-promoting activity. The hydrophobic peptide(s) of AAT are also found in human bile, and in a bile model are known to promote cholesterol crystallisation. The aim of this study was to analyse the levels and molecular profile of AAT in gall bladder bile samples obtained during laparoscopic cholecystectomy. Bile samples were obtained from 23 patients with gallstones, 7 of which had cholestasis and 4 had both cholestasis and cholangitis; mean age 59.5 years. The concentration of AAT was estimated by immunoelectrophoresis and molecular forms were determined by 12.5% SDS-PAGE electrophoresis followed with Western Blot analysis. Blots were developed using antisera against AAT or C-terminal fragment of AAT (C-36). We found higher levels of AAT (3.6--0.4 g/L) in 17% and very low levels (0.0(04).08 g/L) in 83% of all samples. The C-36 peptide was found in 9 of 23 analysed samples, and 6 of those had very low concentrations of AAT. Moreover, the peptide was found in patients diagnosed for gallstones but not in subgroups with cholestasis, cholangitis or both. The levels of cleaved AAT correlate with low concentrations of native AAT in gall bladder bile and suggest that the generation of cleaved forms of AAT may affect gallstone formation in vivo.
Background & Aim: It is unclear whether UDCA improves liver injury caused by bifiary obstruction. Therefore, it was the aim to study the effects of UDCA in obstructive cholestasis in mice. Methods: Mice were fed a UDCA (0.5% w/w) supplemented diet or control diet for 7 d. Thereafter mice were subjected to common bile duct ligation (CBDL; n = 24) or selective bile duct ligation (SBDL; n = 12) and continued on UDCA or control diet for further 3 d. In addition, naive mice were subjected to UDCA after CBDL (n = 6. (Furthermore, Mdr2 - / - (with non-suppurative cholangitis) were fed UDCA (0.5% w/w) for 7 d. Mortality rates, serum liver enzymes, total serum bile acid levels, bile composition, and liver histology were investigated. Results: CBDL and SBDL (ligated lobes) resulted in disseminated necrosis, bile infarcts, and periductular fibrosis, whereas non-ligated lobes remained unchanged. CBDL (3 days) in UDCA-fed mice aggravated necrosis and significantly increased mortality rates (7/12 vs 1/12 in controls; p < 0.05). UDCA feeding to SBDL mice aggravated necrosis exclusively in ligated lobes, whereas non-ligated remained unchanged. UDCA feeding started after CBDL resulted in 100% mortality after 5 days (6/6). Moreover, UDCA feeding resulted in bile infarcts in Mdr2 - / - . UDCA feeding had no effect on liver histology and liver enzymes in sham operated animals and wild type mice. Summary & Conclusions: UDCA aggravates liver injury in obstructive cholestasis in mice. Further clinical studies will have to determine potential UDCA side effects in patients with obstructive cholestasis.
~-]
ABNORMAL VASCULAR COMPLIANCE IN PATIENTS WITH PRIMARY SILIARY CIRRHOSIS
I.S.H. Cadden, B.M. Mullah, M.E. Callender, I,S. Young. Dept of Medicine, Royal Victoria Hospital, Belfast, Northern Ireland, UK Introduction: Elevated serum markers have suggested endothelial dysfunction in Primary Biliary Cirrhosis (PBC). We used the technique of Pulse Wave Analysis (PWA) to study the arterial stiffness (increased in endothelial dysfunction) in patients with PBC. PWA uses applanation tonometery to record the radial artery pressure waveform non-invasively. Application of a reverse transfer function allows accurate reconstruction of the central aortic waveform and determination of the central aortic pressures. Augmentation of this pressure thus provides a measurement of the compliance of the vascular tree, and this can be expressed quantitatively as the augmentation index (Alx).
~-]
HEPATIC UPTAKE OF CHOLECYSTOKININ OCTAPEPTIDE (CCK-8) BY ORGANIC ANION TRANSPORTING POLYPEPTIDES OATP4 AND OATP8 OF RAT AND HUMAN LIVER
M.G. Ismair, V. Cattori, B. Hagenbuch, P.J. Meier, G.A. Kullack-Ublick.
Department of Medicine, University Hospital, Switzerland Background & Aims: Cholecystokinin (CCK) is a major gastrointestinal peptide hormone which is released postprandially from the small intestine and which exerts marked effects on gallbladder and gastrointestinal motility. The smaller isoforms CCK-8 and CCK-4 are rapidly taken up into hepatocytes, metabolized and finally excreted into bile. Our aim was to identify and characterize the hepatocellular CCK-8 uptake system.