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Lipid turnover in alcoholics before and after an alcohol load
HEPATOLOGY Vol. 22, No. 4, Pt. 2, 1995
1289
AASLD ABSTRACTS
MRI FINDINGS IN CHRONIC HEPATIC ENCEPHALOPATHY: RESULTS OF A RANDOMIZED TRIAL COMPARING TRANSJUGULAR INTRAHEPATIC PORTOSYSTEMIC SHUNT AND SCLEROTHERAPY. S Herrmann (1L D Krieaer {2), O Jansen f3L M Geil31er(4L P Sauer (1). A Stiehl (1~,.M Jau8 (2L W. Stremmel (1L and L. Theilmann (1L (1) Dept. of Gastroenterology, (2) Dept. of Neurology, (3) Dept. of Neuroradiology, (4) Dept. of Psychiatryof the RuprechtKarls University of Heidelberg, FRG Chronic hepatic encephalopathy (CHE) is a progressive disorder characterized predominantly by mental deteriorat on, desntegrat on of personality, extrapyramidal symptoms, and MRI abnormalities. The aim of this study was to compare development of CHE in patients receiving either TIPS or elective sclerotherapy (EST). 12 patients (8 males) receiving TIPS (45.1 ys + 12 SD, Child 6.2 +_1.g SD) and 12 patients (4 males) receiving EST (54.5 ys + 14.6 SD, Child 6.0 _+1.8 SD) were recruited from an ongoing prospective randomized trial evaluating TIPS versus EST for the treatment of portal hypedension. Repeat cranial MRI (T1weighted MRI to determine paitidal hyperintensity in relation to frontal white matter), standardized neuropsychiatric evaluation (dichotomized categories for neurologic symptoms, Brief Psychiatric Rating Scale), and psychometric tests (psychomotor speed, verbal and visual memory, and concentration) were applied before and 6 months after TIPS or EST respectively. Test results were analysed for changes using Wilcoxon rank sum test for two independent samples. At study entry groups did not differ in any chosen criteria. TIPS remained patent during study period with mean pressure gradients below 20 mmHg. At follow-up, pallidal hyperintensity increased significantly (p<.01) along with neuropsychiatric deterioration (p<.05) in TIPS patients. Psychometric testing could not distinguish between EST and TIPS after 6 months. CHE worsens with TIPS in comparison to EST. Psychometric tests did not prove helpful in monitoring progessive CHE. Therapeutic desicions towards TIPS have to consider current neuropsychiatric performance and MRI findings for the patient's benefit.
1291 LIPID TURNOVER IN
ALCOHOLICS BEFORE AND AFTER AN ALCOHOL LOAD, Sandra Hirsch. M. Pia de la Maza, Margarita Petermann. Daniel Bunout. Institute of Nutrition and Food Technology and Faculty of Medicine, University of Chile. San Borja Arriar/,n Hospital. Alcohol ingestion decreases plasma free fatty acids (FFA) and lipid oxidation. The aim of this investigation .was to study palmitate turnover in alcoholics during a short abstinence period and after an ethanol load, and in a group of non-alcoholic controls, looking for correlations between palmitate turnover, FFA, acetate and acetoacetate/beta hydroxibutarate ratio. Patients and methods: Palmitate CI4 turnover was studied in 5 alcoholics during early abstinence and after a 0.8 g/kg ethanol load, and in 5 non alcoholic normal cont~'ols.Plasma levels of FFA, acetate, acetoacetate and beta hydroxybutirate were measured before and during the ethanol load. A needle hepatic biopsy was obtained in alcoholics. Results: FFA levels, palmitate flux, oxidation and non-oxidative disposal were similar in alcoholics compared to controls, decreasing significantly after the ethanol load in both groups. A positive correlation was found between FFA levels and palmitate flux. Liver biopsies showed mild changes in the patients studied. Conclusions: The similar inhibition of lipid turnover and FFA release observed after an alcohol load in alcoholics and control subjects suggests that this effectis mediated by alcohol metabolism and not by metabolic alterations present in alcoholics. •
Financing: Fondecyt, Chile Grant # 1930951.
,,
V.
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429A
1290 REPLICATION-DEFECTIVE HBV MUTANTS IN AN IMMUNOLOGICALLY NEGATIVE PATIENT. B. Hiller, H.-J. Schlayer, T. Peters, J. Fehr, H.E.Blum and J. Rasenack. Dept. of Medicine, AlbertLudwigs-University, Freiburg, Germany. HBV DNA was amplified by PCR from the serum of a patient who participated in a post-transfusion hepatitis study and who was negative for HBsAg, HBeAg, anti-HBs, anti-HBc and anti-HB¢. Overlapping fragments were cloned and at least ten clones of each fragment sequenced. Sequence comparison with all published HBV genomes revealed 22 constant nucleic acid exchanges, which were found in all clones sequenced and 38 inconstant mutations which were found only in a portion of the clones sequenced indicating a mixed HBV population. One of these inconstant mutations was an 8 bp deletion in the C-prom0tor leading to a truncation ~of the X-protein. Another inconstant mutation in the C promotor was a 1 bp insertion leading to an X-C fusion-protein. To test the biological relevance of these mutations, genomes were constructed which contained: A) all constant mutations and further nine inconstant mutations assembled from original PCR fragments; B) only all constant mutations; C) an NcoI-MroI fragment from A) cloned into wild type; D) a PCR fragment containing the deletion cloned into wild type; E) a PCR fragment containing the insertion cloned into wild type. These constructs were transfected as head-to-tail dimers into Huh7 cells. HBsAg expression was almost normal in all mutants except from A). HBeAg expression was reduced to zero in mutant B, to 30 % in D, to 50 % in mutant E. HBV-DNA replication was absent in mutants A, B and D and significantly reduced in mutant E. The results indicate that different mutations may contribute to the diminished rate of replication and expression in immunologically negative patients.
1292 EFFECT OF PENTOXIFYLLINE IN A HETEROLOGOUS SERUM MODEL OF FIBROSIS IN THE RAT. P.D. Hodgson and T.C. Peterson : Dept of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada. Fibrosis occurs as an imbalance between collagen deposition and removal. Hepatic fibrosis is characterized by fibroblast proliferation and increased collagen synthesis. Pentoxifylline (PTX) inhibits (in a dose related manner) in vitro PDGF stimulated fibrublast proliferation (Peterson et al 1994). Yellow phosphorus induced fibrosis in the pig can be blocked with 4 weeks of PTX (16 mg/kg p.o.) (Peterson, 1993). Recently we investigated a heterologous serum induced model of fibrosis in the rat. The treatment group (n=6) received swine serum (SS) (0.5 ml i.p. 2x weekly) (n=6), while the control group (n=4) received saline (0.5 ml i.p. 2x weekly). ALT was significantly elevated in the SS treated group after 16 weeks of treatment (control 18.362 0.36, SS 23.28_+ 1.31 p=0.02), at which point the SS group was divided into 2 sub groups (n=3). One subgroup also received PTX (32 mg/kg/day i.p. 5 days/wk) in addition to SS. No significant difference in ALT was observed between PTX and SS groups after 6 weeks PTX (control 162 1, PTX+SS 26.4_+ 1.4, SS 33.36_+ 4.56 IU/1). Liver section collagen estimation by Sirius Red/Fast Green, obtained at 6 weeks FIX, showed no difference between PTX and SS groups (control 10.1_+ 0.5, PTX+SS 15.7_+0.5, SS 13.26_+ 0.6 ~tg/mg protein). Histological studies of liver sections stained by Massons trichrome supported this data. 3H-thymidine uptake by rat fibroblasts stimulated with monocyte conditioned media from SS treated rats (5735_+ 540 cpm) indicated no significant difference in fibroproliferation compared to controls (5489_+ 817 cpm), suggesting that this model of fibrosis is not a fibmproliferative one. Further studies have shown that PTX treatment does not lower elevated PIII NP values. In conclusion PTX is not capable of reversing the heterologous serum model of fibrosis in the rat. (supported by MRC Canada)
1289
AASLD ABSTRACTS
MRI FINDINGS IN CHRONIC HEPATIC ENCEPHALOPATHY: RESULTS OF A RANDOMIZED TRIAL COMPARING TRANSJUGULAR INTRAHEPATIC PORTOSYSTEMIC SHUNT AND SCLEROTHERAPY. S Herrmann (1L D Krieaer {2), O Jansen f3L M Geil31er(4L P Sauer (1). A Stiehl (1~,.M Jau8 (2L W. Stremmel (1L and L. Theilmann (1L (1) Dept. of Gastroenterology, (2) Dept. of Neurology, (3) Dept. of Neuroradiology, (4) Dept. of Psychiatryof the RuprechtKarls University of Heidelberg, FRG Chronic hepatic encephalopathy (CHE) is a progressive disorder characterized predominantly by mental deteriorat on, desntegrat on of personality, extrapyramidal symptoms, and MRI abnormalities. The aim of this study was to compare development of CHE in patients receiving either TIPS or elective sclerotherapy (EST). 12 patients (8 males) receiving TIPS (45.1 ys + 12 SD, Child 6.2 +_1.g SD) and 12 patients (4 males) receiving EST (54.5 ys + 14.6 SD, Child 6.0 _+1.8 SD) were recruited from an ongoing prospective randomized trial evaluating TIPS versus EST for the treatment of portal hypedension. Repeat cranial MRI (T1weighted MRI to determine paitidal hyperintensity in relation to frontal white matter), standardized neuropsychiatric evaluation (dichotomized categories for neurologic symptoms, Brief Psychiatric Rating Scale), and psychometric tests (psychomotor speed, verbal and visual memory, and concentration) were applied before and 6 months after TIPS or EST respectively. Test results were analysed for changes using Wilcoxon rank sum test for two independent samples. At study entry groups did not differ in any chosen criteria. TIPS remained patent during study period with mean pressure gradients below 20 mmHg. At follow-up, pallidal hyperintensity increased significantly (p<.01) along with neuropsychiatric deterioration (p<.05) in TIPS patients. Psychometric testing could not distinguish between EST and TIPS after 6 months. CHE worsens with TIPS in comparison to EST. Psychometric tests did not prove helpful in monitoring progessive CHE. Therapeutic desicions towards TIPS have to consider current neuropsychiatric performance and MRI findings for the patient's benefit.
1291 LIPID TURNOVER IN
ALCOHOLICS BEFORE AND AFTER AN ALCOHOL LOAD, Sandra Hirsch. M. Pia de la Maza, Margarita Petermann. Daniel Bunout. Institute of Nutrition and Food Technology and Faculty of Medicine, University of Chile. San Borja Arriar/,n Hospital. Alcohol ingestion decreases plasma free fatty acids (FFA) and lipid oxidation. The aim of this investigation .was to study palmitate turnover in alcoholics during a short abstinence period and after an ethanol load, and in a group of non-alcoholic controls, looking for correlations between palmitate turnover, FFA, acetate and acetoacetate/beta hydroxibutarate ratio. Patients and methods: Palmitate CI4 turnover was studied in 5 alcoholics during early abstinence and after a 0.8 g/kg ethanol load, and in 5 non alcoholic normal cont~'ols.Plasma levels of FFA, acetate, acetoacetate and beta hydroxybutirate were measured before and during the ethanol load. A needle hepatic biopsy was obtained in alcoholics. Results: FFA levels, palmitate flux, oxidation and non-oxidative disposal were similar in alcoholics compared to controls, decreasing significantly after the ethanol load in both groups. A positive correlation was found between FFA levels and palmitate flux. Liver biopsies showed mild changes in the patients studied. Conclusions: The similar inhibition of lipid turnover and FFA release observed after an alcohol load in alcoholics and control subjects suggests that this effectis mediated by alcohol metabolism and not by metabolic alterations present in alcoholics. •
Financing: Fondecyt, Chile Grant # 1930951.
,,
V.
-
429A
1290 REPLICATION-DEFECTIVE HBV MUTANTS IN AN IMMUNOLOGICALLY NEGATIVE PATIENT. B. Hiller, H.-J. Schlayer, T. Peters, J. Fehr, H.E.Blum and J. Rasenack. Dept. of Medicine, AlbertLudwigs-University, Freiburg, Germany. HBV DNA was amplified by PCR from the serum of a patient who participated in a post-transfusion hepatitis study and who was negative for HBsAg, HBeAg, anti-HBs, anti-HBc and anti-HB¢. Overlapping fragments were cloned and at least ten clones of each fragment sequenced. Sequence comparison with all published HBV genomes revealed 22 constant nucleic acid exchanges, which were found in all clones sequenced and 38 inconstant mutations which were found only in a portion of the clones sequenced indicating a mixed HBV population. One of these inconstant mutations was an 8 bp deletion in the C-prom0tor leading to a truncation ~of the X-protein. Another inconstant mutation in the C promotor was a 1 bp insertion leading to an X-C fusion-protein. To test the biological relevance of these mutations, genomes were constructed which contained: A) all constant mutations and further nine inconstant mutations assembled from original PCR fragments; B) only all constant mutations; C) an NcoI-MroI fragment from A) cloned into wild type; D) a PCR fragment containing the deletion cloned into wild type; E) a PCR fragment containing the insertion cloned into wild type. These constructs were transfected as head-to-tail dimers into Huh7 cells. HBsAg expression was almost normal in all mutants except from A). HBeAg expression was reduced to zero in mutant B, to 30 % in D, to 50 % in mutant E. HBV-DNA replication was absent in mutants A, B and D and significantly reduced in mutant E. The results indicate that different mutations may contribute to the diminished rate of replication and expression in immunologically negative patients.
1292 EFFECT OF PENTOXIFYLLINE IN A HETEROLOGOUS SERUM MODEL OF FIBROSIS IN THE RAT. P.D. Hodgson and T.C. Peterson : Dept of Pharmacology, Dalhousie University, Halifax, Nova Scotia, Canada. Fibrosis occurs as an imbalance between collagen deposition and removal. Hepatic fibrosis is characterized by fibroblast proliferation and increased collagen synthesis. Pentoxifylline (PTX) inhibits (in a dose related manner) in vitro PDGF stimulated fibrublast proliferation (Peterson et al 1994). Yellow phosphorus induced fibrosis in the pig can be blocked with 4 weeks of PTX (16 mg/kg p.o.) (Peterson, 1993). Recently we investigated a heterologous serum induced model of fibrosis in the rat. The treatment group (n=6) received swine serum (SS) (0.5 ml i.p. 2x weekly) (n=6), while the control group (n=4) received saline (0.5 ml i.p. 2x weekly). ALT was significantly elevated in the SS treated group after 16 weeks of treatment (control 18.362 0.36, SS 23.28_+ 1.31 p=0.02), at which point the SS group was divided into 2 sub groups (n=3). One subgroup also received PTX (32 mg/kg/day i.p. 5 days/wk) in addition to SS. No significant difference in ALT was observed between PTX and SS groups after 6 weeks PTX (control 162 1, PTX+SS 26.4_+ 1.4, SS 33.36_+ 4.56 IU/1). Liver section collagen estimation by Sirius Red/Fast Green, obtained at 6 weeks FIX, showed no difference between PTX and SS groups (control 10.1_+ 0.5, PTX+SS 15.7_+0.5, SS 13.26_+ 0.6 ~tg/mg protein). Histological studies of liver sections stained by Massons trichrome supported this data. 3H-thymidine uptake by rat fibroblasts stimulated with monocyte conditioned media from SS treated rats (5735_+ 540 cpm) indicated no significant difference in fibroproliferation compared to controls (5489_+ 817 cpm), suggesting that this model of fibrosis is not a fibmproliferative one. Further studies have shown that PTX treatment does not lower elevated PIII NP values. In conclusion PTX is not capable of reversing the heterologous serum model of fibrosis in the rat. (supported by MRC Canada)