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686 Comparison of two clinical scales in the causality assessment of drug induced liver disease
08. Alcoholic liver disease, NAFLD and drug induced liver disease ethanol 20% sucrose 20% (w/v) solution as the only drinking fluid, or an isocaloric diet without alcohol. Livers were fixed in paraformaldehyde for histology or flash-frozen for biochemical analysis. AST and ALT in peripheral blood were determined by routine procedure. Vitamin E content in liver homogenates was assessed by HPLC. MnSOD content in liver homogenates and HEK-293 cell protein lysates was determined by immunoblotting. Results: Although WT and p66 KO mice drank comparable amounts of ethanol, treatment resulted in WT mice in a marked increase of ALT and AST in the bloodstream and in a significant reduction of liver Vitamin E content, consistent with the establishment of hepatocyte oxidative injury, while these changes were markedly attenuated in p66 KO animals. Importantly, liver steatosis and organ swelling were observed in WT mice, but were nearly absent in p66 KO. Resistance to ethanol toxicity by p66 deletion correlated with an higher expression of the mitochondrial Superoxide dismutase (MnSOD) in liver tissue. Moreover, overexpression of p66 in HEK-293 cells led to a significant downregulation of MnSOD in this cell line, confirming that mitochondrial defenses are negatively regulated by p66shc in a cellautonomous fashion. Conclusions: The above results clearly indicate that deletion of p66shc protects mice from alcoholic liver damage, concomitantly with an increase in mitochondrial antioxidant defenses. This observation strongly supports a role for p66 and ROS in ethanol toxicity. Moreover, MnSOD downregulation by p66 may represent a general link between p66 deletion, resistance to oxidative stress and longevity.
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COMPARISON OF TWO CLINICAL SCALES IN THE CAUSALITY A S S E S S M E N T OF DRUG INDUCED LIVER DISEASE
M. Garcla-Cort6sl., K. Pachkoria 3, M.I. Lucena2, R. Andrade 1, R. Camargo 1, R. Alcantara 1, R. Hidalgo4. 1Department of
Gastroenterology and Hepatology, 2Department of Clinical Pharmacology, Hospital Universitario Virgen de la Victoria, Malaga, Spain," 3Department of Pharmacology, School of Medicine, Malaga, Spain," 4Statistic Centre, University of Malaga, Spain Introduction: Diagnosis of drug induced liver disease is difficult since no specific tests are available. In order to improve causality assessment different methods have been developed. We have noticed differences between authors and referees in the results yielded by the Naranjo scale required by an International Journal in the evaluation of cases of hepatotoxicity submitted for publication. Objectives: The aim of this study was to compare two diagnostic methods: The Naranjo adverse drug reaction scale (non-specific) and the Council for International Organizations of Medical Sciences (CIOMS) scale (liver specific) in order to elucidate the validity of the former in the diagnosis of hepatotoxicity. Methods: 225 cases of suspected drug induced liver disease submitted to a registry were evaluated applying both scales by two independent raters. 32 of those cases were not finally included in the registry but were used in the study as a control group. A total of 249 ratings were generated due to a double suspected drug in 24 patients. Consistency was evaluated by the weighted kappa statistical test. Results: The Naranjo scale showed a sensitivity of 54%, specificity of 88%, and 95% positive and 29% negative predictive value. Besides, the majority of the scores yielded by this scale were in the midrange with a very low number of definitive or excluded results showing a low discriminative power. The grade of agreement between observers with the Naranjo scale was low (44%) with a 0.23 weighted kappa, compared with a concordance of 73% with a 0.87 weighted kappa when applying the CIOMS scale. Agreement between scales was 24% with a 0.29 weighted kappa. Conclusions: The Naranjo scale showed a poor validity and reproducibility and comes to be an unspecific method for the evaluation of hepatotoxic
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reactions; therefore we recommend the application of the CIOMS scale in cases of suspected drug induced liver disease not only in the pharmacovigilance setting and by medical journals but also in the daily clinical practice.
~8--~ PLATELET-ACTIVATING FACTOR INACTIVATOR ENHANCES LIVER'S RECOVERY FROM PARACETAMOL (ACETAMINOPHEN) INTOXICATION A.D. Grypioti 1'2, G. Kostopanagiotou3, G.K. Papadimas 1, C.A. Demopoulos2, M.G. Mykoniatis 1. 1Department of Experimental
Pharmacology, Medical School, 2Department of Chemistry, National and Kapodistrian University of Athens, Greece," 32nd Department of Anaesthesiology, School of Medicine, Attikon Hospital, University of Athens, Greece Background and Alms: Hepatic outcome after paracetamol (acetaminophen) intoxication is compromised by tissue injury and release of hepatic enzymes and by production of pathological mediators. The PlateletActivating Factor (PAF) is an endogenous lipid-mediator, which plays a key role in catalysing various pro-inflammatory processes associated with acute liver failure. The aim of this study was to investigate the effects of PAF inactivator, recombinant PAF-acetylhydrolase (rPAF-AH) on post paracetamol-treatment functional outcome of the liver in the rat. Methods: Fifty male Wistar rats were divided into two groups: the control group received by gastric tube a toxic dose of paracetamol (3.5 g/kg BW) suspended in saline solution and the rPAF-AH-treated group received the same dose of paracetamol followed by a dose of rPAF-AH (5 mg/kg BW) intraperitoneally. The animals were sacrificed at time points of 20, 24, 28, 32 and 40h after paracetamol treatment. The hepatic injury was evaluated at the different times interval after intoxication by determination of serum enzyme activities of AST, ALT, ALP and LDH. Hepatic levels of malondialdehyde (MDA), serum cholesterol/HDL cholesterol fraction and serum superoxide dismutase (SOD) activity were also measured as indicators of tissue damage and as parameters of oxidant-antioxidant balance. Results: In control group of rats paracetamol was found to cause oxidative stress followed by acute hepatic injury. Compared with the control group, the addition of PAF inactivator had a significant effect in tissue recovery. The positive effects of rPAF-AH were expressed by: (1) reduction of oxidative stress evident by low levels of MDA and cholesterol/HDL cholesterol fraction and increase of SOD activity and (2) high decrease of hepatic injury as showed by all serum enzyme activities (AST, ALT, ALP and LDH) measured. Conclusions: The use of PAF inactivator enhances liver's recovery from paracetamol intoxication, rPAF-AH attenuates the severity of experimental liver injury and can provide important means of improving liver function following paracetamol intoxication. These results suggest that PAF plays an important role in the pathogenesis of paracetamol-induced liver injury and may act as a signaling mediator in the initiation and amplification of the hepatic inflammatory process.
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NON-INVASIVE TESTS OF LIVER FIBROSIS IN NAFLD: A SYSTEMATIC REVIEW
I.N. Guhal~ J. Parkes 2, RR. Roderick 2, W.M. Rosenberg1. 1University
of Southmapton Liver Unit, 2public Health Seiences and Statistics, Southmapton University, Southampton, UK Background: Non-alcoholic fatty liver disease (NAFLD) has a spectrum of disease from simple steatosis to cirrhosis. Long term studies suggest that the presence of fibrosis within non-alcoholic fatty liver disease (NAFLD) has a considerable prognostic implication. Currently liver biopsy is the gold standard diagnostic test for liver fibrosis but due to its limitations there is a need to develop effective non-invasive tests.
•
COMPARISON OF TWO CLINICAL SCALES IN THE CAUSALITY A S S E S S M E N T OF DRUG INDUCED LIVER DISEASE
M. Garcla-Cort6sl., K. Pachkoria 3, M.I. Lucena2, R. Andrade 1, R. Camargo 1, R. Alcantara 1, R. Hidalgo4. 1Department of
Gastroenterology and Hepatology, 2Department of Clinical Pharmacology, Hospital Universitario Virgen de la Victoria, Malaga, Spain," 3Department of Pharmacology, School of Medicine, Malaga, Spain," 4Statistic Centre, University of Malaga, Spain Introduction: Diagnosis of drug induced liver disease is difficult since no specific tests are available. In order to improve causality assessment different methods have been developed. We have noticed differences between authors and referees in the results yielded by the Naranjo scale required by an International Journal in the evaluation of cases of hepatotoxicity submitted for publication. Objectives: The aim of this study was to compare two diagnostic methods: The Naranjo adverse drug reaction scale (non-specific) and the Council for International Organizations of Medical Sciences (CIOMS) scale (liver specific) in order to elucidate the validity of the former in the diagnosis of hepatotoxicity. Methods: 225 cases of suspected drug induced liver disease submitted to a registry were evaluated applying both scales by two independent raters. 32 of those cases were not finally included in the registry but were used in the study as a control group. A total of 249 ratings were generated due to a double suspected drug in 24 patients. Consistency was evaluated by the weighted kappa statistical test. Results: The Naranjo scale showed a sensitivity of 54%, specificity of 88%, and 95% positive and 29% negative predictive value. Besides, the majority of the scores yielded by this scale were in the midrange with a very low number of definitive or excluded results showing a low discriminative power. The grade of agreement between observers with the Naranjo scale was low (44%) with a 0.23 weighted kappa, compared with a concordance of 73% with a 0.87 weighted kappa when applying the CIOMS scale. Agreement between scales was 24% with a 0.29 weighted kappa. Conclusions: The Naranjo scale showed a poor validity and reproducibility and comes to be an unspecific method for the evaluation of hepatotoxic
$253
reactions; therefore we recommend the application of the CIOMS scale in cases of suspected drug induced liver disease not only in the pharmacovigilance setting and by medical journals but also in the daily clinical practice.
~8--~ PLATELET-ACTIVATING FACTOR INACTIVATOR ENHANCES LIVER'S RECOVERY FROM PARACETAMOL (ACETAMINOPHEN) INTOXICATION A.D. Grypioti 1'2, G. Kostopanagiotou3, G.K. Papadimas 1, C.A. Demopoulos2, M.G. Mykoniatis 1. 1Department of Experimental
Pharmacology, Medical School, 2Department of Chemistry, National and Kapodistrian University of Athens, Greece," 32nd Department of Anaesthesiology, School of Medicine, Attikon Hospital, University of Athens, Greece Background and Alms: Hepatic outcome after paracetamol (acetaminophen) intoxication is compromised by tissue injury and release of hepatic enzymes and by production of pathological mediators. The PlateletActivating Factor (PAF) is an endogenous lipid-mediator, which plays a key role in catalysing various pro-inflammatory processes associated with acute liver failure. The aim of this study was to investigate the effects of PAF inactivator, recombinant PAF-acetylhydrolase (rPAF-AH) on post paracetamol-treatment functional outcome of the liver in the rat. Methods: Fifty male Wistar rats were divided into two groups: the control group received by gastric tube a toxic dose of paracetamol (3.5 g/kg BW) suspended in saline solution and the rPAF-AH-treated group received the same dose of paracetamol followed by a dose of rPAF-AH (5 mg/kg BW) intraperitoneally. The animals were sacrificed at time points of 20, 24, 28, 32 and 40h after paracetamol treatment. The hepatic injury was evaluated at the different times interval after intoxication by determination of serum enzyme activities of AST, ALT, ALP and LDH. Hepatic levels of malondialdehyde (MDA), serum cholesterol/HDL cholesterol fraction and serum superoxide dismutase (SOD) activity were also measured as indicators of tissue damage and as parameters of oxidant-antioxidant balance. Results: In control group of rats paracetamol was found to cause oxidative stress followed by acute hepatic injury. Compared with the control group, the addition of PAF inactivator had a significant effect in tissue recovery. The positive effects of rPAF-AH were expressed by: (1) reduction of oxidative stress evident by low levels of MDA and cholesterol/HDL cholesterol fraction and increase of SOD activity and (2) high decrease of hepatic injury as showed by all serum enzyme activities (AST, ALT, ALP and LDH) measured. Conclusions: The use of PAF inactivator enhances liver's recovery from paracetamol intoxication, rPAF-AH attenuates the severity of experimental liver injury and can provide important means of improving liver function following paracetamol intoxication. These results suggest that PAF plays an important role in the pathogenesis of paracetamol-induced liver injury and may act as a signaling mediator in the initiation and amplification of the hepatic inflammatory process.
•
NON-INVASIVE TESTS OF LIVER FIBROSIS IN NAFLD: A SYSTEMATIC REVIEW
I.N. Guhal~ J. Parkes 2, RR. Roderick 2, W.M. Rosenberg1. 1University
of Southmapton Liver Unit, 2public Health Seiences and Statistics, Southmapton University, Southampton, UK Background: Non-alcoholic fatty liver disease (NAFLD) has a spectrum of disease from simple steatosis to cirrhosis. Long term studies suggest that the presence of fibrosis within non-alcoholic fatty liver disease (NAFLD) has a considerable prognostic implication. Currently liver biopsy is the gold standard diagnostic test for liver fibrosis but due to its limitations there is a need to develop effective non-invasive tests.