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Lidocaine metabolite formation in chronic liver diseases
498A
AASLD ABSTRACTS
1565 NOVELORN1THINE DECARBOXYLASE PROTEIN IN HUMAN HEPATOCELLULARCARCINOMA. A Tamori. S Nishiguchi, N Koh. T Kuroki. Y Yano, S Otani. H Kinoshita, and K Kobavashi. Third Dept. of Internal Medicine, Second Dept. of Biochemistry, and Second Dept. of Surgery, Osaka City University Medical School, Osaka 545, Japan. Ornithine decarboxylase (ODC) is important in tumor promotion and its half-life is within an one hour. We have reported that its activity is correlated with the differentiation of hepatocellular carcinoma (HCC). There are some reports that cancers have a mutant ODC protein. We examined the sequence of ODC cDNA from resected liver of three patients with HCC. Products of the reverse transcription and polymerase chain reaction from ODC m R N A of HCC and noncancerous tissues were sequenced and the amino acid sequences were deduced. There were three point-mutations with amino acid replacement in HCC: codon 415 glutamic acid to a stop codon, codon 431 glutamie acid to glycine, and codon 435 serine to glycine. These mutant ODCs were synthesized with an in vitro retieulocyte lysate transcription-translation system. The stability of synthesized proteins was examined by means of an ATP reticulocyte-degradation system. The activity of truncated ODC remained in high level after 3 h incubation. These results show that HCC has an ODC that differs from that of normal tissue. lh
2h
3h
100%
Oh
34 %
19 %
14 %
MutarL ODC 100% (trune~ed-tTpe)
78%
60%
60%
Wild-type
1566
HEPATOLOGY O c t o b e r 1995
NEONATAL HEPATITIS OF NON-FAMILIAL FORM ASSOCIATED WITH FAT AND HEMOSIDERIN DEPOSIT IN THE LIVER OF BREAST-FED JAPANESE
INFANTS. Y Tazawa, F Nishinumiya, D Abukama*, J Aikawa*, M Nakagawa*, F Nishinomiya, G Takada. T Konno*. Dept. of Pediatrics, University of Akita, Akita, and "University of Tohoku, Seedai. We describe five neonatal hepatitis patients, 2 males and 3 females, 34 to 70 days, presenting an unpublished clinical features. All 5 patients were all full-term, normal-birthweight and breast-fed, and no mother had received any medication after birth. Jaundice (5/5), hepatumegaly (5/5; 2-5 ca) and failure to thrive (2/5; weight gain, 18-19 g/day) were major findings on physical examination. Laboratory data were summarized as follows: Total bilirubin {4.8-15.8 mg/dl), direct bilirubln (1.9-11.2 mg/dl), Hgb (7.9-9.5 g/dl), GOT (28-
175 IU/L), GPT (25-45 IU/L), GGTP (142=205 IU/L), ALP (5912,230 IU/L), galaetosemia (24-99 mg/dl), hyperferritinemla (n= 2, 1,459-1,791 ng/ml), low activities of thrumbotest (n=3, 2225Z), hypertyrosinumia with a negative r e s u l t for urinary suecinylacetone (n=2), normal activities of galactose-l-phosphate urldyl transferase, galactokinase and uridine diphosphate galaetase 4-epimerase in red blood cells (n=2), normal levels of serum earn±thine levels (n=2), and fatty-fibrotie patterns on ultrasound tomography (n=2). No patients had metabolic acidosis, hyperammenamia and hypoglycemia, well delineated biliary traetviral and other metabolic diseases mere eliminated by pertinent tests. Liver biopsy disclosed moderate to severe fatty liver associated with hemesiderin deposit in both hepatic and retieuloendothelial cells (n=5). Bucenl mecnsal biopsy was negative for iron deposit {n=1). After subsltating breast milk for lactose-free formula, liver dysfunction became normal within 4 to 8 months of age. A11 patients are alive with normal liver tests (I-17 yrs), although subsequent liver biopies showed mild fibrosis at the age of 1 to 2 yrs (n=2). In conclusion, we present an unrecognized disease in early infancy, mimiking galaetosemia or neonatal hamoehromatasis with survival. The cause of this disease may be related with breast milk.
1567 THE RELATIONSHIP BETWEEN HEPATITIS B VIRAL GENOTYPE,
1568 LIDOCAINE METABOLITE FORMATION IN C H R O N I C
PRE-CORE MUTATIONS, ENCAPSIDATION SIGNAL STABILITY, OVERALL R E P L I C A T I O N RATE AND H I S T O L O G I C A L ABNORMALITIES IN anti-HBeAg-POSITIVE PATIENTS F ter Borg, M Wester*, HTM Cuvpers*, FW ten Kate1", RAFM Chamulean, HW Reesink, A. Leentvaar± Liver Unit and ~'Dep. of Pathology, Acad. Med. Center of the University of Amsterdam, *CLB and :~GG&GD, Amsterdam, the Netherlands
LIVER DISEASES. R Testa, N Campo, L. Arzani, S Alvarez, S. Caglieris, D Risso, PB Lantieri, G Celle. DI.M.I., Gastroenterology Unit, University of Genoa, Italy. Monoethylglycinexylidide (MEGX) serum levels at 15, 30 or 60 min after i.v. lidocaine bolus, were considered to be a quantitative liver function test. The aim of this work was to compare the MEG)( formation among 130 patients with chronic active hepatitis (CAH, 30 F, 100 M, age 16 + 73) and 154 patients with cirrhosis (CIR, 46 F, 108 M, age 28 + 73). Of these patients, 45 were mild CAH, 54 moderate CAH, 31 CAH with cirrhosis (CAHcir), 49 CIR A, 78 CIR B and 27 CIR C (Child-Pugh's grading). MEGX at 15, 30 and 60 min after i.v. lidocaine (1 mg/Kg), was measured by TDX fluorescent polarization immunoassay. The differences in the MEGX means (±SD, ng/ml) between progressive degrees of liver damage were statistically analyzed by means of ANOVA (*p < 0.05). Discriminant level (L0) between CAH and C/R patients was calculated for every sampling time and sensitivity (SS) and specificity (SP) were computed. At each sampling time MEGX levels showed a decrease according to the progression of liver damage: MEGX 15 MEGX 30 MEGX 60 CAH mild 74 ± 30 82 ± 25 77 ± 20 CAH moderate 65±30 * 77±26 75 :~ 21 * CAH cir 35±14" 48±15" 52±14" CIRA 26±21 36±23 * 39±22 * C1RB 13± 8* 21±12' 26±12" CIRC 5± 8 13±11 13±10" The discriminant level between CAH and CIR of MEGX 15 (31 ng/ml) showed 91.5 SS and 84.6 in SP, of MEGX 30 (44 ng/ml) 87.6 in SS and 82.3 in SP, and of MEGX 60 (47 ng/ml) 90.2 in SS and 84.6 in SP. These results showed that MEGX formation are able to evaluate functional derangement of progressive liver diseases and can be useful to differentiate between chronic active hepatitis and cirrhosis.
Factors influencing pathogenicity of the hepatitis B virus (HBV) in anti-HBeAgpositive patients include genotype, pre-core mutations and overall replicative activity. Replication may be modulated by mutations affecting the stability of a tertiary structure in the pre-core segment of the pre-genomic HBV-RNA (e-loop), which may be responsible for uptake of HBV-RNA in the nascent core-particle. We prospectively investigated these variables in 65 patients (52% with normal ALT-values), all of whom underwent liver biopsy. The Knodell score and a score of inflammatory activity were applied blindly to all biopsies. Serum HBV-DNA concentration ([HBV-DNA]) was measured by the branchedDNA hybridization assay (positive when [HBV-DNA] exceeds 0.7"106 copies/ml) and by a quantitative polymerase chain reaction assay (positive when [HBV-DNA] exceeds -700 copies/ml). If HBV-DNA was demonstrable (55 cases), it was subjected to nucleic acid sequencing, permitting classification into genotypes (which were classified as A or non-A) and pre-core mutant or wildtype virus. In addition, the stability of the e-loop was calculated (entropy, AG). Log[HBV-DNA] was found to be correlated with inflammatory activity score (r =0.58), Knodell score (r=0.59) and ALT-level (r=0.45) (p's <0.0001). AG also correlated with Iog[HBV-DNA] (r=0.28,p =0.03), but the correlation between AG and inflammatory scores did not reach statistical significance (r=0.23, p=0.09). Mean AG was lower in genotype A (26.6) than in genntypes non-A (28.5) (p<0.001), while pre-core mutations in general were more frequent in genotypes non-A (79%) than in genotype A (38%) (p =0.004). These findings suggest that non-A HBV genotypes are more prone to pre-core mutations and in general have higher stability of the e-loop than the A-genotype. The e-loop stability seems to be positively correlated with viral replication, which is strongly linked to hepatic inflammatory activity in chronic anti-HBeAg-positive hepatitis B.
AASLD ABSTRACTS
1565 NOVELORN1THINE DECARBOXYLASE PROTEIN IN HUMAN HEPATOCELLULARCARCINOMA. A Tamori. S Nishiguchi, N Koh. T Kuroki. Y Yano, S Otani. H Kinoshita, and K Kobavashi. Third Dept. of Internal Medicine, Second Dept. of Biochemistry, and Second Dept. of Surgery, Osaka City University Medical School, Osaka 545, Japan. Ornithine decarboxylase (ODC) is important in tumor promotion and its half-life is within an one hour. We have reported that its activity is correlated with the differentiation of hepatocellular carcinoma (HCC). There are some reports that cancers have a mutant ODC protein. We examined the sequence of ODC cDNA from resected liver of three patients with HCC. Products of the reverse transcription and polymerase chain reaction from ODC m R N A of HCC and noncancerous tissues were sequenced and the amino acid sequences were deduced. There were three point-mutations with amino acid replacement in HCC: codon 415 glutamic acid to a stop codon, codon 431 glutamie acid to glycine, and codon 435 serine to glycine. These mutant ODCs were synthesized with an in vitro retieulocyte lysate transcription-translation system. The stability of synthesized proteins was examined by means of an ATP reticulocyte-degradation system. The activity of truncated ODC remained in high level after 3 h incubation. These results show that HCC has an ODC that differs from that of normal tissue. lh
2h
3h
100%
Oh
34 %
19 %
14 %
MutarL ODC 100% (trune~ed-tTpe)
78%
60%
60%
Wild-type
1566
HEPATOLOGY O c t o b e r 1995
NEONATAL HEPATITIS OF NON-FAMILIAL FORM ASSOCIATED WITH FAT AND HEMOSIDERIN DEPOSIT IN THE LIVER OF BREAST-FED JAPANESE
INFANTS. Y Tazawa, F Nishinumiya, D Abukama*, J Aikawa*, M Nakagawa*, F Nishinomiya, G Takada. T Konno*. Dept. of Pediatrics, University of Akita, Akita, and "University of Tohoku, Seedai. We describe five neonatal hepatitis patients, 2 males and 3 females, 34 to 70 days, presenting an unpublished clinical features. All 5 patients were all full-term, normal-birthweight and breast-fed, and no mother had received any medication after birth. Jaundice (5/5), hepatumegaly (5/5; 2-5 ca) and failure to thrive (2/5; weight gain, 18-19 g/day) were major findings on physical examination. Laboratory data were summarized as follows: Total bilirubin {4.8-15.8 mg/dl), direct bilirubln (1.9-11.2 mg/dl), Hgb (7.9-9.5 g/dl), GOT (28-
175 IU/L), GPT (25-45 IU/L), GGTP (142=205 IU/L), ALP (5912,230 IU/L), galaetosemia (24-99 mg/dl), hyperferritinemla (n= 2, 1,459-1,791 ng/ml), low activities of thrumbotest (n=3, 2225Z), hypertyrosinumia with a negative r e s u l t for urinary suecinylacetone (n=2), normal activities of galactose-l-phosphate urldyl transferase, galactokinase and uridine diphosphate galaetase 4-epimerase in red blood cells (n=2), normal levels of serum earn±thine levels (n=2), and fatty-fibrotie patterns on ultrasound tomography (n=2). No patients had metabolic acidosis, hyperammenamia and hypoglycemia, well delineated biliary traetviral and other metabolic diseases mere eliminated by pertinent tests. Liver biopsy disclosed moderate to severe fatty liver associated with hemesiderin deposit in both hepatic and retieuloendothelial cells (n=5). Bucenl mecnsal biopsy was negative for iron deposit {n=1). After subsltating breast milk for lactose-free formula, liver dysfunction became normal within 4 to 8 months of age. A11 patients are alive with normal liver tests (I-17 yrs), although subsequent liver biopies showed mild fibrosis at the age of 1 to 2 yrs (n=2). In conclusion, we present an unrecognized disease in early infancy, mimiking galaetosemia or neonatal hamoehromatasis with survival. The cause of this disease may be related with breast milk.
1567 THE RELATIONSHIP BETWEEN HEPATITIS B VIRAL GENOTYPE,
1568 LIDOCAINE METABOLITE FORMATION IN C H R O N I C
PRE-CORE MUTATIONS, ENCAPSIDATION SIGNAL STABILITY, OVERALL R E P L I C A T I O N RATE AND H I S T O L O G I C A L ABNORMALITIES IN anti-HBeAg-POSITIVE PATIENTS F ter Borg, M Wester*, HTM Cuvpers*, FW ten Kate1", RAFM Chamulean, HW Reesink, A. Leentvaar± Liver Unit and ~'Dep. of Pathology, Acad. Med. Center of the University of Amsterdam, *CLB and :~GG&GD, Amsterdam, the Netherlands
LIVER DISEASES. R Testa, N Campo, L. Arzani, S Alvarez, S. Caglieris, D Risso, PB Lantieri, G Celle. DI.M.I., Gastroenterology Unit, University of Genoa, Italy. Monoethylglycinexylidide (MEGX) serum levels at 15, 30 or 60 min after i.v. lidocaine bolus, were considered to be a quantitative liver function test. The aim of this work was to compare the MEG)( formation among 130 patients with chronic active hepatitis (CAH, 30 F, 100 M, age 16 + 73) and 154 patients with cirrhosis (CIR, 46 F, 108 M, age 28 + 73). Of these patients, 45 were mild CAH, 54 moderate CAH, 31 CAH with cirrhosis (CAHcir), 49 CIR A, 78 CIR B and 27 CIR C (Child-Pugh's grading). MEGX at 15, 30 and 60 min after i.v. lidocaine (1 mg/Kg), was measured by TDX fluorescent polarization immunoassay. The differences in the MEGX means (±SD, ng/ml) between progressive degrees of liver damage were statistically analyzed by means of ANOVA (*p < 0.05). Discriminant level (L0) between CAH and C/R patients was calculated for every sampling time and sensitivity (SS) and specificity (SP) were computed. At each sampling time MEGX levels showed a decrease according to the progression of liver damage: MEGX 15 MEGX 30 MEGX 60 CAH mild 74 ± 30 82 ± 25 77 ± 20 CAH moderate 65±30 * 77±26 75 :~ 21 * CAH cir 35±14" 48±15" 52±14" CIRA 26±21 36±23 * 39±22 * C1RB 13± 8* 21±12' 26±12" CIRC 5± 8 13±11 13±10" The discriminant level between CAH and CIR of MEGX 15 (31 ng/ml) showed 91.5 SS and 84.6 in SP, of MEGX 30 (44 ng/ml) 87.6 in SS and 82.3 in SP, and of MEGX 60 (47 ng/ml) 90.2 in SS and 84.6 in SP. These results showed that MEGX formation are able to evaluate functional derangement of progressive liver diseases and can be useful to differentiate between chronic active hepatitis and cirrhosis.
Factors influencing pathogenicity of the hepatitis B virus (HBV) in anti-HBeAgpositive patients include genotype, pre-core mutations and overall replicative activity. Replication may be modulated by mutations affecting the stability of a tertiary structure in the pre-core segment of the pre-genomic HBV-RNA (e-loop), which may be responsible for uptake of HBV-RNA in the nascent core-particle. We prospectively investigated these variables in 65 patients (52% with normal ALT-values), all of whom underwent liver biopsy. The Knodell score and a score of inflammatory activity were applied blindly to all biopsies. Serum HBV-DNA concentration ([HBV-DNA]) was measured by the branchedDNA hybridization assay (positive when [HBV-DNA] exceeds 0.7"106 copies/ml) and by a quantitative polymerase chain reaction assay (positive when [HBV-DNA] exceeds -700 copies/ml). If HBV-DNA was demonstrable (55 cases), it was subjected to nucleic acid sequencing, permitting classification into genotypes (which were classified as A or non-A) and pre-core mutant or wildtype virus. In addition, the stability of the e-loop was calculated (entropy, AG). Log[HBV-DNA] was found to be correlated with inflammatory activity score (r =0.58), Knodell score (r=0.59) and ALT-level (r=0.45) (p's <0.0001). AG also correlated with Iog[HBV-DNA] (r=0.28,p =0.03), but the correlation between AG and inflammatory scores did not reach statistical significance (r=0.23, p=0.09). Mean AG was lower in genotype A (26.6) than in genntypes non-A (28.5) (p<0.001), while pre-core mutations in general were more frequent in genotypes non-A (79%) than in genotype A (38%) (p =0.004). These findings suggest that non-A HBV genotypes are more prone to pre-core mutations and in general have higher stability of the e-loop than the A-genotype. The e-loop stability seems to be positively correlated with viral replication, which is strongly linked to hepatic inflammatory activity in chronic anti-HBeAg-positive hepatitis B.