- Email: [email protected]
Assesment of Albumin Binding Capacity (ABiC) for characterization of albumin transport function by a new method
Category 8: Nutrition, metabolism, alcoholic liver disease, pharmacology followed by 2 injections at 3-month intervals, respectively). The primary end point was serum HBV DNA negativation at 6 and 12 months by using the Cobas HBV-PCR test (cut-off value = 200 copies/ml). Results: At inclusion, only 3/28 subjects had undetectable viremia by PCR. In the 25 others, the viral load was low (<100.000 in all but 1 with 238.000 copies/ml). At month 6 and 12, there was no significant variation in the quantitative HBV load or in the relative rate of HBV DNA decrease by comparison with baseline values between non vaccinated (33%) and vaccinated subjects (35%). There was no variation according to the route of immunization. None developed transaminase flare-up, immune complexes disease or antiHBs. One vaccinated subject cleared HBsAg 26 months after vaccination. Conclusions: This trial shows that: 1. most of healthy HBsAg carriers has still detectable low levels of HBV replication; 2. specific HBV vaccine therapy does not cancel this residual replication.
-~
ASSESMENT OF ALBUMIN BINDING CAPACITY (ABiC) FOR CHARACTERIZATION OF ALBUMIN TRANSPORT FUNCTION BY A NEW METHOD
S. Klammt, B. Brinkmann, S.R. Mitzner, J. Stange, J. Loock, E. Peters, J. Emmrich, S. Liebe, E. Munzert. University of Rostock, Dept. of
Internal Medicine, Gastroenterology., Inst.for Clinical Chemistry and Pathobiochemistry., TERAKLINAG, Rostock, Germany Estimation of albumin concentration is clinically used as a liver function test and as a marker of severity of illnes in multiple scoring systems and therefore as a predictive prognostic value. However, measuring only concentration of albumin does not provide informations about the present transport function or capacity for lipophilic endogenous ligands (e.g. prostaglandins, bilirubin, bile acids) or drugs (e.g. diazepam, diuretics). Considering the need of an easy to handle estimation of existing albumin transport function especially in liver disease a new laboratory test has been developed which enables the binding site specific characterization of the Albumin Binding Capacity (ABiC). Methods: After incubating different albumin samples with a binding site II (diazepam binding site) specific fluorescence marker (Dansylsarcosine, DS) the amount of unbound DS can be determined by fluorescence detection after ultrafiltration. In parallel, the same procedure is done with a standard albumin for reference. Thus the binding capacity of the diazepam binding site can be quantified. Results: With increasing site II specific albumin ligand concentration up to a molar ratio of 2:1 ABiC is reduced (by 48.8% for N-acetyltryptophan or by 52.8% for diazepam, respectively), whereas increasing concentrations of ligands bound to other sites like bilirubin do not effect ABiC. In first clinical evaluations a significant reduction of ABiC in patients with cholestasis could be shown (44.2 4- 13% compared to an ABiC of 105.6 4- 30% in healty volunteers). Conclusions: Measuring Albumin Binding Capacity (ABiC) by a new laboratory test offers the opportunity to characterize severity of liver disease not only by albumin concentration but also by assesment of transport function. Clinical trials are ongoing to investigate the diagnostic potential of this new method.
~-~
CYTOCHROME P4502E1 EXPRESSION IN PATIENTS WITH ALCOHOLIC AND NON-ALCOHOLIC STEATOHEPATITIS
J. Li 1 , H. Cortez-Pinto 2 , R. Waldherr 1 , M. Ingelman-Sundberg 3 , A. Baptista 2, M.E. Camilo 2, H.K. Seitz 1. i Dept. of Medicine, Salem
Medical Center, Heidelberg, Germany; 2Dept. of Med., Univ. Hosp. Santa Maria, Lisboa, Portugal; 3Dept, of Med. Biochem. and Biophysics, Karolinska Inst., Stockholm, Sweden Chronic ethanol consumption results in an induction of hepatic cytochrome P4502E1 (CYP2E1). This induction leads to the production of free radicals during ethanol metabolism and it has been proposed as
197
one important mechanism for alcoholic liver disease (ALD). In addition, CYP2E1 induction also occurs in non-alcoholic steatohepatitis (NASH) (Weltmann et al, Hepatology 27: 128, 1998). To evaluate CYP2E1 induction in alcoholic steatohepatitis (ASH) and in NASH and its role in the pathogenesis of liver disease immunohistochemistry was performed in biopsies from 8 patients with ASH and 14 patients with NASH. In all cases CYP2E1 induction was centrolobular and in some also midzonal induction was observed. One out of 6 patients with ASH had a moderate induction of CYP2E1 while the remaining patients had a slight induction only. Three out of 14 NASH patients had a moderate induction of CYP2E1 while the remaining patients had a slight or no induction at all. CYP2E1 induction did not correlate with other morphological parameters such as necrosis, inflammation or fibrosis, nor with the occurrence of Mallory bodies. It is interesting that one NASH patient with cirrhosis of the liver did not show any signal with respect to CYP2E1, suggesting that CYP2E1 induction may operate only in the first stages of the disease. In conclusion, CYP2E1 is induced in ASH and NASH but did not correlate with other morphological parameters of liver disease. Thus, the degree of CYP2E1 induction is not a prognostic maker of NASH and does not reflect the severity of the disease.
~-2~
APOLIPOPROTEIN E POLYMORPHISM IN PATIENTS WITH NONAL-COHOLIC STEATOHEPATITIS IN A MEXICAN POPULATION
H. Rodriguez-Hernandez 1, B. Ruiz-madrigal 2, J. Luis Gonzalez 1, G. Martinez Aguilar 1, M.R. Reyes 3, A. Panduro-Cerda 2.1 I.M.S.S.,
Durango; 2 CUCS UdeG, Guadalajara; 3 CLAN, Faculty of Medicine UJED, Durango, Mexico Aim: To study correlations between apo E polymorphism and NASH. Setting: Hospital of the Mexican Institute of Social Security in Durango, Mexico. Design and Methods: Transversal comparative study. Three men and 27 women with BMI index > 27 kg/m2 and histological evidence of NASH were included. Paired controls were 5 and 25 health adult men and women respectively, with BMI < 27 kg/m2. Blood chemistry, liver function tests, lipid profile and plasma malondialdehyde (MDA) were determined. Apo E alleles were determined by PCR and restriction mapping. Statistical analysis was done with the Mann-Whitney test. Results and Conclusions: Statistically significant differences (p < 0.05) were observed between both groups in BMI (33.1 4- 4.8 kg/m2 vs 23.9 4- 1.2 kg/m2), glucose (1122.7 4- 27.5 mg/dL vs 85 4- 2 mg/dL), AST (105.2 4- 14.6 U/L vs 24.2 4- 3.7 U/L), ALT (111.4 4- 15.2 U/L vs 26.4 4- 9.5 U/L), AP (136.8 4- 17 U/L vs 85.2 4- 14.7 U/L), cholesterol (191.8 4- 47.2 mg/dL vs 177 4- 22.3 mg/dL), triglycerides (334 4- 87.8 mg/dL vs 112.9 4- 43.7 mg/dL) and MDA (89.8 4- 23 nmol/L vs 14.05 4- 6.5 nmol/L). In the NASH group severe fibrosis or chirrhosis were not detected. The results showed almost similar epsilon 3 and 4 apo E allele frequencies between NASH patients and controls, only epsilon 2 allele was underrepresented in NASH patients. More studies in this issue are warranted since the high prevalence of this disorder. (Grant ~ ( ~ 3 8 / 8 1 4 from I.M.S.S. to HRH).
~-3-~ HEME-ARGINATE IN THE TREATMENT OF ACUTE PORPHYRIAS : EFFECT ON IRON METABOLISM AND HEME CATABOLITES J.P. Deybach, H.V. Puy, Y.A. Nordmann. Biochemistry and Molecular
Gentics, Centre Francais des Porphyries, Hopital Louis Mourier, France We have investigated the effect of heme-arginate administered intravenously in the treatment of 856 acute attacks in 172 porphyric patients. Iron status (iron, ferritin, transferrin serum levels and serum transferrinsaturation) and heme catabolites produced by heme oxygenase activity
-~
ASSESMENT OF ALBUMIN BINDING CAPACITY (ABiC) FOR CHARACTERIZATION OF ALBUMIN TRANSPORT FUNCTION BY A NEW METHOD
S. Klammt, B. Brinkmann, S.R. Mitzner, J. Stange, J. Loock, E. Peters, J. Emmrich, S. Liebe, E. Munzert. University of Rostock, Dept. of
Internal Medicine, Gastroenterology., Inst.for Clinical Chemistry and Pathobiochemistry., TERAKLINAG, Rostock, Germany Estimation of albumin concentration is clinically used as a liver function test and as a marker of severity of illnes in multiple scoring systems and therefore as a predictive prognostic value. However, measuring only concentration of albumin does not provide informations about the present transport function or capacity for lipophilic endogenous ligands (e.g. prostaglandins, bilirubin, bile acids) or drugs (e.g. diazepam, diuretics). Considering the need of an easy to handle estimation of existing albumin transport function especially in liver disease a new laboratory test has been developed which enables the binding site specific characterization of the Albumin Binding Capacity (ABiC). Methods: After incubating different albumin samples with a binding site II (diazepam binding site) specific fluorescence marker (Dansylsarcosine, DS) the amount of unbound DS can be determined by fluorescence detection after ultrafiltration. In parallel, the same procedure is done with a standard albumin for reference. Thus the binding capacity of the diazepam binding site can be quantified. Results: With increasing site II specific albumin ligand concentration up to a molar ratio of 2:1 ABiC is reduced (by 48.8% for N-acetyltryptophan or by 52.8% for diazepam, respectively), whereas increasing concentrations of ligands bound to other sites like bilirubin do not effect ABiC. In first clinical evaluations a significant reduction of ABiC in patients with cholestasis could be shown (44.2 4- 13% compared to an ABiC of 105.6 4- 30% in healty volunteers). Conclusions: Measuring Albumin Binding Capacity (ABiC) by a new laboratory test offers the opportunity to characterize severity of liver disease not only by albumin concentration but also by assesment of transport function. Clinical trials are ongoing to investigate the diagnostic potential of this new method.
~-~
CYTOCHROME P4502E1 EXPRESSION IN PATIENTS WITH ALCOHOLIC AND NON-ALCOHOLIC STEATOHEPATITIS
J. Li 1 , H. Cortez-Pinto 2 , R. Waldherr 1 , M. Ingelman-Sundberg 3 , A. Baptista 2, M.E. Camilo 2, H.K. Seitz 1. i Dept. of Medicine, Salem
Medical Center, Heidelberg, Germany; 2Dept. of Med., Univ. Hosp. Santa Maria, Lisboa, Portugal; 3Dept, of Med. Biochem. and Biophysics, Karolinska Inst., Stockholm, Sweden Chronic ethanol consumption results in an induction of hepatic cytochrome P4502E1 (CYP2E1). This induction leads to the production of free radicals during ethanol metabolism and it has been proposed as
197
one important mechanism for alcoholic liver disease (ALD). In addition, CYP2E1 induction also occurs in non-alcoholic steatohepatitis (NASH) (Weltmann et al, Hepatology 27: 128, 1998). To evaluate CYP2E1 induction in alcoholic steatohepatitis (ASH) and in NASH and its role in the pathogenesis of liver disease immunohistochemistry was performed in biopsies from 8 patients with ASH and 14 patients with NASH. In all cases CYP2E1 induction was centrolobular and in some also midzonal induction was observed. One out of 6 patients with ASH had a moderate induction of CYP2E1 while the remaining patients had a slight induction only. Three out of 14 NASH patients had a moderate induction of CYP2E1 while the remaining patients had a slight or no induction at all. CYP2E1 induction did not correlate with other morphological parameters such as necrosis, inflammation or fibrosis, nor with the occurrence of Mallory bodies. It is interesting that one NASH patient with cirrhosis of the liver did not show any signal with respect to CYP2E1, suggesting that CYP2E1 induction may operate only in the first stages of the disease. In conclusion, CYP2E1 is induced in ASH and NASH but did not correlate with other morphological parameters of liver disease. Thus, the degree of CYP2E1 induction is not a prognostic maker of NASH and does not reflect the severity of the disease.
~-2~
APOLIPOPROTEIN E POLYMORPHISM IN PATIENTS WITH NONAL-COHOLIC STEATOHEPATITIS IN A MEXICAN POPULATION
H. Rodriguez-Hernandez 1, B. Ruiz-madrigal 2, J. Luis Gonzalez 1, G. Martinez Aguilar 1, M.R. Reyes 3, A. Panduro-Cerda 2.1 I.M.S.S.,
Durango; 2 CUCS UdeG, Guadalajara; 3 CLAN, Faculty of Medicine UJED, Durango, Mexico Aim: To study correlations between apo E polymorphism and NASH. Setting: Hospital of the Mexican Institute of Social Security in Durango, Mexico. Design and Methods: Transversal comparative study. Three men and 27 women with BMI index > 27 kg/m2 and histological evidence of NASH were included. Paired controls were 5 and 25 health adult men and women respectively, with BMI < 27 kg/m2. Blood chemistry, liver function tests, lipid profile and plasma malondialdehyde (MDA) were determined. Apo E alleles were determined by PCR and restriction mapping. Statistical analysis was done with the Mann-Whitney test. Results and Conclusions: Statistically significant differences (p < 0.05) were observed between both groups in BMI (33.1 4- 4.8 kg/m2 vs 23.9 4- 1.2 kg/m2), glucose (1122.7 4- 27.5 mg/dL vs 85 4- 2 mg/dL), AST (105.2 4- 14.6 U/L vs 24.2 4- 3.7 U/L), ALT (111.4 4- 15.2 U/L vs 26.4 4- 9.5 U/L), AP (136.8 4- 17 U/L vs 85.2 4- 14.7 U/L), cholesterol (191.8 4- 47.2 mg/dL vs 177 4- 22.3 mg/dL), triglycerides (334 4- 87.8 mg/dL vs 112.9 4- 43.7 mg/dL) and MDA (89.8 4- 23 nmol/L vs 14.05 4- 6.5 nmol/L). In the NASH group severe fibrosis or chirrhosis were not detected. The results showed almost similar epsilon 3 and 4 apo E allele frequencies between NASH patients and controls, only epsilon 2 allele was underrepresented in NASH patients. More studies in this issue are warranted since the high prevalence of this disorder. (Grant ~ ( ~ 3 8 / 8 1 4 from I.M.S.S. to HRH).
~-3-~ HEME-ARGINATE IN THE TREATMENT OF ACUTE PORPHYRIAS : EFFECT ON IRON METABOLISM AND HEME CATABOLITES J.P. Deybach, H.V. Puy, Y.A. Nordmann. Biochemistry and Molecular
Gentics, Centre Francais des Porphyries, Hopital Louis Mourier, France We have investigated the effect of heme-arginate administered intravenously in the treatment of 856 acute attacks in 172 porphyric patients. Iron status (iron, ferritin, transferrin serum levels and serum transferrinsaturation) and heme catabolites produced by heme oxygenase activity