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Quantitative analysis of wild-type and precore mutant HBV in the natural history and interferon therapy of chronic hepatitis B
502A
1581
AASLD
THE EFFECT OF GENDER ON GALLBLADDER MOTILITY IN OBESE AND NON-OBESE SUBJECTS. T Trouillot. C McKinley. R Showalter. G Everson. Section of Hepatology and Hepatobiliary Research Center, Dept. of Medicine, General Clinical Research Center, University of Colorado Health Sciences Center, Denver, CO.
ABSTRACTS
1582
THE PATHOLOGY OF THE LIVER GRAFTS AFTER LIVER/SMALL BOWEL OR MULTIVISCERAL TRANSPLANTATION. A C Tsamandas!, K Abu-Elma~d2, H Furukawa2~__STodo2_, AJ Demetris~RQRG Lee ~'. Departments of Pathologyt and Surgery2, Division of Transplantation, University of Pittsburgh Medical Center, Pittsburgh PA, 15261. Background/Aims: Liver/small bowel (LSB) or multivisceral (MV) transplantation is a therapeutic option for patients with short gut syndranre complicated by parenteral nutrition-related end-stage liver disease. We assessed the pathology of these combined liver allografts in comparison with isolated liver allografts. Patients and methods: 51 liver biopsy specimens from 13 patients who underwent LSB or MV transplantation were studied. The control group consisted of 51 specimens from isolated liver grafts matched for diagnosis and patient age. Results: In general, the histologic findings in the LSB or MV groups mirrored those seen with isolated grafts, but prominent portal and Iobular neutrophilia was seen in 47 speeiarens. Although in 15 specimens this was an expected finding associated with sepsis (n=l 1), ischemia/preservation injury (n=3), and bile duet stricture (n=l), it was also seen with acute cellular rejection (ACR) in 10 specimens from 6 patients. Bacteremia was established in 4 patients with the same organism ira blood, stool, and in 1 ease, liver cultures; only 1 allograft small bowel biopsy demonstrated rejection at the sanre time. By eantrast, neutropbilia was not a notable feature in tim cantrol biopsies with ACR or minor non-specific changes; the latter were accompanied by negative blood cultures. Acquired hepatitis C with a "fibrosing-eholestatie" pattern was identified ira 10 biopsies from 2 patiants. Conclusions: The pathology of the liver allografts from the LSB or MV recipients does not differ significantly from that of the isolated liver allograft. Portal and Iobular neutrophilia is a common background feature, and although of variable etiology, may reflect ongoing inflammatory process in the grafted intestine or bacterial translocation. Hepatitis C with a clmlestatie picture may result from immuansappression with modulation of viral genome and possible direct eytotoxieity.
1584
ALTERED REGULATION AND ROLE OF DIPEPTIDYL PEPTIDASE IV DURING HEPATIC REGENERATION. M Tsugiki, Y Kobavashi, T Kawasaki, and T Yoshimi. Second Department of Internal Medicine. Hamamatsu University School of MediCine, Shizuoka, Japan.
Female gender and obesity are risk factors for cholesterol gallstone formation. Existing data suggest that the morbidly obese have defects in gallbladder motility and a significantly larger gallbladder fasting volume that may contribute to cholesterol gallstone formation. For these masons, we determined whether there were gender differences in gallbladder motility in 15 obese subjects. Subjects: 27 healthy obese subjects were screened; 6 were excluded for gallstones (22%), and 6 were excluded for other reasons. None were diabetic or on medications. The characteristics of the study population were: age 38~9, B/vli 35±3, M:F gender 7:8. Measurements were compared to those of matched normal-weight controls. Methods: Fasting volume (FV), residual volume (RV), and fractional emptying (EF) in response to a morning meal containing 15g of fat and 470 calories were determined using real-time ultrasonography. Results: • WOMEN I MEN OBESE NON-OBESE p-value [OBESE NON-OBESE n-value BMl(kg/m 2) 34-+.3 22-d=3 0.000081 36±4 26~2 0.0003 FV(ml) 27"10 15:t:4 0.015 16.d=4 0.009 RV(ml) 10~5 7±4 NS I 12±6 7:t:4 0.07 EF(%) 63*9 56±13 NS I 60~:12 61~-12 NS
130-~9
Gender did not influence gallbladder motility in either obese or non-obese groups. Obesity is characterized by a high prevalence of cholelithiasis and enlarged fasting gallbladder volumes but normal fractional emptying in response to a meal. Conclusions: The female preponderance for cholesterol gallstones is not attributable to gender differences in gallbladder motility. Enlarged gallbladder volumes in the obese may contribute to cholelithinsis by promoting bile stasis. (Partially supported by a grant from Hoff-mann-LaRoche)
1 5 8 3 QUANTITATIVE ANALYSIS OF WILD-TYPE AND PRECOREMUTANT HBV IN THE NATURALHISTORYANDINTERFERONTHERAPYOFCHRONICHEPATITIS B. H Tsugeno. G Yamada. H Endo. F Kishi, M Takatani, M Takahashl. K Manabe, M Kinoshita* and T Tsuji. F i r s t Dept. of I n t e r n a l Medeeine. 0kayama University Medical School. 0kayama. and *Diagnostic Research I n s t i t u t e , Diagnostics Division, 0tsuka Pharmaceutical Co..Ltd.. Tokushima. Japan. The c l i n i c a l s i g n i f i c a n c e of the precore mutant HBV (nt. 1896) in long term outcome of chronic HBV i n f e c t i o n i s s t i l l unclear. A r e l a t i o n between response to interferonlIFNI and the prevalence of wild-type v . s . procure mutant HBV i s s t i l l controv e r s i a l . To c l a r i f y these points, we examined q u a n t i f i c a t i o n of c i r c u r a t i n g wild-type and preeore mutant HBV populations in asymptomatie c a r r i e r s and chronic h e p a t i t i s p a t i e n t s by competitive m u t a t i o n - s i t e - s p e c i f i c assay(CMSSA). Materials and Methods: Serum samples were obtained from asymptamatic healthy c a r r i e r s (ASC)(HBeAg+: n:7. anti-HBe+: n=19) and p a t i e n t s with chronic hepatitis(CH)(HBeAg*: n=7. anti-HBe+:n=6). In addition, amounts of wild-type and procure mutant HBV were analyzed s e r i a l l y in 4 p a t i e n t s t r e a t e d with IFN. HBV-DNAfrom s e r a was amplified by MSSA using d i f f e r e n t s e t s of s p e c i f i c primers. which were complementary only to w i l d - t y p e or to the mutant DNA, a n d q u a n t i f i e d by competitive PCR using an i n t e r n a l contorol. Results: Average amounts of the w i l d - t y p e / p r e c o r e mutant HBV were 2 . 3 x 10g(DNA copies/ml)/3.5x 108(DNA eopies/ml} in HBeAg+ kSC. 2.8xl0S/l. O x l 0 s in HBekg+ CH, 8.04x106/2:3x107 in anti-HRe+ CH. 2.4xi0/I. ix l0 s in anti-HBe÷ ASC. respectively. The ratio of wild-type v.s. precore mutant HBV was 8.1(3~13) in HBeAg÷ ASC. 3.5(1~I0) in HBeAg+ UH. 0.5(0. i~I} in anti-HBe+CH, 0.2(0~I} in anti-HBe÷ ASC. Two of four patients treated with IFN were responders. In response to IFN, amounts of both wild-type and precore mutant HBV were decreased equivalently either in responders or nonresponders. Conolusions: In all HBeAg* ASC and CH. both wild-type and precore mutant IIBV are detected by CMSSA, but in some anti-HBe+ ASC only preeore mutant HBV are detected. There seams to be no signlfieant difference in response to IFN between wild-type and preeore mutant HBV.
HEPATOLOGY O c t o b e r 1 9 9 5
Dipeptidyl peptidase IV (DPPIV) is a bile eanalicular ectopeptidase. In some cellular systems, DPPIV has been implicated in the control of cell growth and differentiation by regulating the local extracellular concentration of biologically active peptides through proteolysis. However. the potential role of DPPIV in liver growth has not been evaluated. Aim: The aim of th e current study was to examine the regulation and role of DPPIV during hepatic regeneration after partial hepatectomy in the rat. Methods: Serum and hepatic DPPIV were serially quantified using an enzymatic assay and binding inhibition assay with anti-DPPIV antibody (mAb HAM.4) in rats undergoing liver regeneration following 2/3 partial bepatectomy. Some animals received cycloheximide to inhibit protein synthesis• Hepatic localization of DPPIV was assessed using an indirect immunoperoxidase technique• The N-terminal amino acid sequence of serum DPPIV was determined by Edman degradation. To evaluate the role of DPPIV in hepatic regeneration, restoration of liver Weight and BrdU incorporation into hepatic DNA were measured after partial hepatectomy in DPPIV-negative and -positive rats. Results: After partial hepatectomy, the amount and activity of DPP1V increased in serum, without any detectable changes in DPPIV within the regenerating liver. Administration of cycloheximide did not inhibit the increase in serum DPPIV after partial hepatectomy. Immunohistochemical analysis showed that the expression of DPPIV was not altered appreciably in the regenerating livers. The N-terminal amino acid sequence of serum DPPIV corresponded to residues beginning at the N-terminal 37th amino acid residue (serine) of the membrane-bound form. There were no significant differences in the restoration of liver weight and hepatic DNA synthesis after partial hepatectomy between DPPIV-negative and DPPIVpositive rats. Conclusions: During hepatic regeneration, the increase in serum DPPIV is not mediated by de hove synthesis of the protein, but may be derived from proteolytic cleavage of the membrane-bound form. DPPIV is not essential for hepatic regeneration in the rat.
1581
AASLD
THE EFFECT OF GENDER ON GALLBLADDER MOTILITY IN OBESE AND NON-OBESE SUBJECTS. T Trouillot. C McKinley. R Showalter. G Everson. Section of Hepatology and Hepatobiliary Research Center, Dept. of Medicine, General Clinical Research Center, University of Colorado Health Sciences Center, Denver, CO.
ABSTRACTS
1582
THE PATHOLOGY OF THE LIVER GRAFTS AFTER LIVER/SMALL BOWEL OR MULTIVISCERAL TRANSPLANTATION. A C Tsamandas!, K Abu-Elma~d2, H Furukawa2~__STodo2_, AJ Demetris~RQRG Lee ~'. Departments of Pathologyt and Surgery2, Division of Transplantation, University of Pittsburgh Medical Center, Pittsburgh PA, 15261. Background/Aims: Liver/small bowel (LSB) or multivisceral (MV) transplantation is a therapeutic option for patients with short gut syndranre complicated by parenteral nutrition-related end-stage liver disease. We assessed the pathology of these combined liver allografts in comparison with isolated liver allografts. Patients and methods: 51 liver biopsy specimens from 13 patients who underwent LSB or MV transplantation were studied. The control group consisted of 51 specimens from isolated liver grafts matched for diagnosis and patient age. Results: In general, the histologic findings in the LSB or MV groups mirrored those seen with isolated grafts, but prominent portal and Iobular neutrophilia was seen in 47 speeiarens. Although in 15 specimens this was an expected finding associated with sepsis (n=l 1), ischemia/preservation injury (n=3), and bile duet stricture (n=l), it was also seen with acute cellular rejection (ACR) in 10 specimens from 6 patients. Bacteremia was established in 4 patients with the same organism ira blood, stool, and in 1 ease, liver cultures; only 1 allograft small bowel biopsy demonstrated rejection at the sanre time. By eantrast, neutropbilia was not a notable feature in tim cantrol biopsies with ACR or minor non-specific changes; the latter were accompanied by negative blood cultures. Acquired hepatitis C with a "fibrosing-eholestatie" pattern was identified ira 10 biopsies from 2 patiants. Conclusions: The pathology of the liver allografts from the LSB or MV recipients does not differ significantly from that of the isolated liver allograft. Portal and Iobular neutrophilia is a common background feature, and although of variable etiology, may reflect ongoing inflammatory process in the grafted intestine or bacterial translocation. Hepatitis C with a clmlestatie picture may result from immuansappression with modulation of viral genome and possible direct eytotoxieity.
1584
ALTERED REGULATION AND ROLE OF DIPEPTIDYL PEPTIDASE IV DURING HEPATIC REGENERATION. M Tsugiki, Y Kobavashi, T Kawasaki, and T Yoshimi. Second Department of Internal Medicine. Hamamatsu University School of MediCine, Shizuoka, Japan.
Female gender and obesity are risk factors for cholesterol gallstone formation. Existing data suggest that the morbidly obese have defects in gallbladder motility and a significantly larger gallbladder fasting volume that may contribute to cholesterol gallstone formation. For these masons, we determined whether there were gender differences in gallbladder motility in 15 obese subjects. Subjects: 27 healthy obese subjects were screened; 6 were excluded for gallstones (22%), and 6 were excluded for other reasons. None were diabetic or on medications. The characteristics of the study population were: age 38~9, B/vli 35±3, M:F gender 7:8. Measurements were compared to those of matched normal-weight controls. Methods: Fasting volume (FV), residual volume (RV), and fractional emptying (EF) in response to a morning meal containing 15g of fat and 470 calories were determined using real-time ultrasonography. Results: • WOMEN I MEN OBESE NON-OBESE p-value [OBESE NON-OBESE n-value BMl(kg/m 2) 34-+.3 22-d=3 0.000081 36±4 26~2 0.0003 FV(ml) 27"10 15:t:4 0.015 16.d=4 0.009 RV(ml) 10~5 7±4 NS I 12±6 7:t:4 0.07 EF(%) 63*9 56±13 NS I 60~:12 61~-12 NS
130-~9
Gender did not influence gallbladder motility in either obese or non-obese groups. Obesity is characterized by a high prevalence of cholelithiasis and enlarged fasting gallbladder volumes but normal fractional emptying in response to a meal. Conclusions: The female preponderance for cholesterol gallstones is not attributable to gender differences in gallbladder motility. Enlarged gallbladder volumes in the obese may contribute to cholelithinsis by promoting bile stasis. (Partially supported by a grant from Hoff-mann-LaRoche)
1 5 8 3 QUANTITATIVE ANALYSIS OF WILD-TYPE AND PRECOREMUTANT HBV IN THE NATURALHISTORYANDINTERFERONTHERAPYOFCHRONICHEPATITIS B. H Tsugeno. G Yamada. H Endo. F Kishi, M Takatani, M Takahashl. K Manabe, M Kinoshita* and T Tsuji. F i r s t Dept. of I n t e r n a l Medeeine. 0kayama University Medical School. 0kayama. and *Diagnostic Research I n s t i t u t e , Diagnostics Division, 0tsuka Pharmaceutical Co..Ltd.. Tokushima. Japan. The c l i n i c a l s i g n i f i c a n c e of the precore mutant HBV (nt. 1896) in long term outcome of chronic HBV i n f e c t i o n i s s t i l l unclear. A r e l a t i o n between response to interferonlIFNI and the prevalence of wild-type v . s . procure mutant HBV i s s t i l l controv e r s i a l . To c l a r i f y these points, we examined q u a n t i f i c a t i o n of c i r c u r a t i n g wild-type and preeore mutant HBV populations in asymptomatie c a r r i e r s and chronic h e p a t i t i s p a t i e n t s by competitive m u t a t i o n - s i t e - s p e c i f i c assay(CMSSA). Materials and Methods: Serum samples were obtained from asymptamatic healthy c a r r i e r s (ASC)(HBeAg+: n:7. anti-HBe+: n=19) and p a t i e n t s with chronic hepatitis(CH)(HBeAg*: n=7. anti-HBe+:n=6). In addition, amounts of wild-type and procure mutant HBV were analyzed s e r i a l l y in 4 p a t i e n t s t r e a t e d with IFN. HBV-DNAfrom s e r a was amplified by MSSA using d i f f e r e n t s e t s of s p e c i f i c primers. which were complementary only to w i l d - t y p e or to the mutant DNA, a n d q u a n t i f i e d by competitive PCR using an i n t e r n a l contorol. Results: Average amounts of the w i l d - t y p e / p r e c o r e mutant HBV were 2 . 3 x 10g(DNA copies/ml)/3.5x 108(DNA eopies/ml} in HBeAg+ kSC. 2.8xl0S/l. O x l 0 s in HBekg+ CH, 8.04x106/2:3x107 in anti-HRe+ CH. 2.4xi0/I. ix l0 s in anti-HBe÷ ASC. respectively. The ratio of wild-type v.s. precore mutant HBV was 8.1(3~13) in HBeAg÷ ASC. 3.5(1~I0) in HBeAg+ UH. 0.5(0. i~I} in anti-HBe+CH, 0.2(0~I} in anti-HBe÷ ASC. Two of four patients treated with IFN were responders. In response to IFN, amounts of both wild-type and precore mutant HBV were decreased equivalently either in responders or nonresponders. Conolusions: In all HBeAg* ASC and CH. both wild-type and precore mutant IIBV are detected by CMSSA, but in some anti-HBe+ ASC only preeore mutant HBV are detected. There seams to be no signlfieant difference in response to IFN between wild-type and preeore mutant HBV.
HEPATOLOGY O c t o b e r 1 9 9 5
Dipeptidyl peptidase IV (DPPIV) is a bile eanalicular ectopeptidase. In some cellular systems, DPPIV has been implicated in the control of cell growth and differentiation by regulating the local extracellular concentration of biologically active peptides through proteolysis. However. the potential role of DPPIV in liver growth has not been evaluated. Aim: The aim of th e current study was to examine the regulation and role of DPPIV during hepatic regeneration after partial hepatectomy in the rat. Methods: Serum and hepatic DPPIV were serially quantified using an enzymatic assay and binding inhibition assay with anti-DPPIV antibody (mAb HAM.4) in rats undergoing liver regeneration following 2/3 partial bepatectomy. Some animals received cycloheximide to inhibit protein synthesis• Hepatic localization of DPPIV was assessed using an indirect immunoperoxidase technique• The N-terminal amino acid sequence of serum DPPIV was determined by Edman degradation. To evaluate the role of DPPIV in hepatic regeneration, restoration of liver Weight and BrdU incorporation into hepatic DNA were measured after partial hepatectomy in DPPIV-negative and -positive rats. Results: After partial hepatectomy, the amount and activity of DPP1V increased in serum, without any detectable changes in DPPIV within the regenerating liver. Administration of cycloheximide did not inhibit the increase in serum DPPIV after partial hepatectomy. Immunohistochemical analysis showed that the expression of DPPIV was not altered appreciably in the regenerating livers. The N-terminal amino acid sequence of serum DPPIV corresponded to residues beginning at the N-terminal 37th amino acid residue (serine) of the membrane-bound form. There were no significant differences in the restoration of liver weight and hepatic DNA synthesis after partial hepatectomy between DPPIV-negative and DPPIVpositive rats. Conclusions: During hepatic regeneration, the increase in serum DPPIV is not mediated by de hove synthesis of the protein, but may be derived from proteolytic cleavage of the membrane-bound form. DPPIV is not essential for hepatic regeneration in the rat.