bluuebunny/crossref_metadata_2025 huggingface dataset:
Transport characteristics of three fluorescent conjugated bile acid analogs in isolated rat hepatocytes and couplets
Authors: Maglova Lilia M. , Jackson Angela M. , Meng Xue-Jun , Carruth Michael W. , Schteingart Claudio D. , Ton-Nu Huong-Thu , Hofmann Alan F. , Weinman Steven A.
Published: March 2007

The transport properties of three different synthetically prepared fluorescent conjugated bile acid analogs (FBA), all with the fluorophore on the side chain, were determined using isolated rat hepatocytes and hepatocyte couplets. The compounds studied were cholylglycylamidofluorescein (CGamF), cholyl-(Nη-nitrobenzoxadiazolyl [NBD])-lysine (C-NBD-L), and chenodeoxycholyl-(Nη-NBD)-lysine (CDC-NBD-L). When hepatocytes were incubated at 37°C with 0.3 μmol/L of FBA and 0.15 mol/L of Na+, cell fluorescence increased linearly with time at a rate (U/min) of 7.8 ± 0.5 for CGamF, 7.2 ± 0.3 for C-NBD-L, and 13.7 ± 1.0 for CDC-NBD-L (mean, ± SE; n = 40 to 90). Uptake was concentration dependent for concentrations less than 20 μmol/L and was saturable. The Michaelis constant (Km) value (μmol/L) for CGamF was 10.8, for C-NBD-L was 3.8, and for CDC-NBD-L was 3.0. In the absence of Na+, the uptake rate was decreased by 50% for CGamF and by 38% for C-NBD-L; but uptake of CDC-NBD-L was unchanged and thus Na+ independent. Cellular uptake of all three derivatives was specific to hepatocytes and was absent in several nonhepatocyte cell lines. For CGamF and C-NBD-L, both Na+-dependent and Na+-independent uptake was inhibited by 200-fold excess concentrations of cholyltaurine, dehydrocholyltaurine, and cholate, but for CDC-NBD-L, these non-fluorescent bile acids did not inhibit initial uptake. The intracellular fluorescence of CGamF was strongly pH dependent at an excitation wavelength of 495 nm, but pH independent at 440 nm excitation. In contrast, intracellular fluorescence of C-NBD-L and CDC-NBD-L was pH independent. All three FBA were secreted into the canalicular space of approximately 50% to 60% of couplets. Cellular adenosine triphosphate (ATP) depletion with either CN− or atractyloside inhibited secretion of all three FBA. The multispecific organic anion transporter (MOAT) inhibitor, chlorodinitro-benzene, blocked secretion of fluorescent MOAT substrates at a concentration of 1 μmol/L. At this concentration it did not affect secretion of the three FBA. At higher concentrations, chlorodinitrobenzene partially inhibited the canalicular secretion of CGamF but not the other two FBA. In conclusion, all three FBA are secreted by canalicular membrane bile acid transporters, but the sinusoidal uptake characteristics of CGamF and C-NBD-L are more similar than those of CDC-NBD-L to the transport properties of cholyltaurine. Therefore, C-NBD-L appears to be the best of the three for studies of conjugated trihydroxy-bile acid transport in hepatocytes. (Hepatology 1995;22:637-647.)
Hepatitis C: IIb (IV) or not IIb (IV) that is the question
Authors: Sallie Richard
Published: March 2007

Objective:

To analyze the distribution of hepatitis C virus (HCV) genotypes among patients positive for antibody to HCV (anti-HCV) according to age, severity of liver disease, and duration of infection; to investigate the influence of HCV genotypes on response to interferon-α therapy; and to study HCV viremia levels in relation to genotypes and severity of liver disease.

Design:

Cross-sectional study.

Setting:

3 university hospitals and 2 research units.

Patients:

3 groups of French and Italian patients with chronic HCV infection and detectable serum HCV RNA: Group 1 included 35 patients with hepatocellular carcinoma; group 2, 71 patients with cirrhosis who did not have hepatocellular carcinoma; and group
Hepatocellular carcinoma complicating autoimmune hepatitis: Role of hepatitis C virus
Authors: Ryder Stephen D. , Koskinas John , Rizzi Paolo M. , McFarlane Ian G. , Portmann Bernard C. , Naoumov Nikolai V. , Williams Roger
Published: March 2007

The risk of hepatocellular carcinoma in autoimmune hepatitis is low, even in patients with long-standing cirrhosis. Because of the increasing recognition of an association of hepatitis C virus (HCV) with autoimmune hepatitis, at least in some geographical areas, and with hepatocellular carcinoma (HCC) (hepatoma), we have examined eight cases (4 male, 4 female) who presented between 1985 and 1993 with hepatoma complicating autoimmune hepatitis. All had steroid-responsive autoimmune hepatitis with serum anti-smooth muscle and antinuclear autoantibodies. Median duration of disease was 17.1 years, and all patients had biopsy-proven cirrhosis. One patient had a history of intravenous drug abuse, and four had previously received blood transfusions. Serum samples (stored at −20°C from up to 9 years before diagnosis of hepatoma) were tested for anti-hepatitis C virus antibodies by a second-third-generation assay and for HCV RNA by the polymerase chain reaction method using primers from the 5′noncoding region. Tissue from liver adjacent to tumor areas was subjected to polymerase chain reaction along with tissue from previous liver biopsy specimens (taken up to 19 years before diagnosis of hepatoma) in all patients. Six patients had evidence of HCV infection: four seropositive for HCV RNA (two of whom were also anti-HCV positive) and two seronegative for HCV RNA and anti-HCV but with HCV RNA in liver tissue at presentation with hepatoma. Retrospective testing showed probable acquisition of HCV through blood transfusion in the four transfused patients. The findings suggest that HCV may have oncogenic potential and that hepatoma complicating autoimmune hepatitis is associated with unsuspected HCV infection in most cases in our series. Screening of liver for HCV RNA should be considered in autoimmune hepatitis patients with a previous history of parenteral exposure to blood, and for hepatoma in those found to have HCV infection. (Hepatology 1995; 22:718-722.)
Rapid and efficient gene transfer in human hepatocytes by herpes viral vectors
Authors: Fong Yuman , Federoff Howard J. , Brownlee Michael , Blumberg David , Blumgart Leslie H. , Brennan Murray F.
Published: March 2007

Retroviral vectors have been widely studied as vehicles for hepatocyte gene therapy, but they are limited by an inability to infect nondividing cells and the need for prolonged cell culture. Two replication deficient herpes simplex viral vectors (HSV) were constructed with the marker genes lac-Z/β-galactosidase (HSVlac) or human-growth hormone (HSVhGH) to determine the efficiency of HSV gene transfer into adult human hepatocytes. Hepatocytes were isolated by collagenase perfusions and density centrifugation from liver wedge biopsy specimens obtained from six patients. After exposure to HSV (0, 50,000, and 500,000 viral particles/106 hepatocytes) for 20 minutes, 1 hour, or 2 hours, the hepatocytes were washed and placed in culture. Hepatocytes transduced with HSVlac were fixed at 24 hours and histochemically stained with X-gal, and media from HSVhGH-transduced cells were assayed at 48 hours by radioimmunoassay for hGH. After a 20-minute exposure at a multiplicity of infection of 0.5 (1 viral particle per 2 hepatocytes), greater than 35% of the hepatocytes expressed the lac-Z gene (>70% efficiency). hGH was also detected in the media from HSVhGH-transduced cells, showing that proteins coded for by foreign genes are not only expressed by transduced cells but are also secreted. Isolated liver perfusions using HSVlac were also performed in Fischer rats. A 20-minute isolated perfusion using 5 × 106 viral particles resulted in expression of the β-galactosidase gene in the rodent livers 72 hours later without histological signs of tissue injury. HSV vectors are potentially powerful tools for gene therapy of human liver disease, because they are efficient and rapid vehicles for gene transfer. Ex vivo modified hepatocytes theoretically may be ready for reinfusion within 100 minutes of liver resection. Efficient in vivo delivery of foreign gene may also be accomplished using these vectors. (Hepatology 1995; 22:723-729.)
Reversal of atrial natriuretic peptide resistance by increasing distal tubular sodium delivery in patients with decompensated cirrhosis
Authors: Abraham William T. , Lauwaars Marije E. , Kim Jin K. , Peñna Ricardo L. , Schrier Robert W.
Published: March 2007

To test the hypothesis that diminished sodium delivery to the distal tubular site of atrial natriuretic peptide (ANP) action accounts for renal ANP resistance in cirrhosis, 12 cirrhotic patients with ascites were studied at baseline and during the infusion of ANP alone (0.15 μg/kg/min), mannitol alone (4 g/hr), and ANP plus mannitol for 3 hours each. Distal tubular sodium delivery, as assessed by lithium clearance, was increased during the infusion of mannitol (13.8 ± 3.4 to 23.7 ± 5.7 mL/min; P < .05) and during the ANP plus mannitol infusion (13.8 ± 3.4 to 28.5 ± 6.3 mL/min; P < .001) in 6 patients, subsequently termed “responders.” Both responders and non-responders were resistant to the natriuretic effect of ANP infused alone, and mannitol alone did not produce an increase in urinary sodium excretion. However, in responders, the mannitol-induced increase in distal tubular sodium delivery resulted in a fivefold increase in urinary sodium excretion during ANP infusion (29 ± 6 to 154 ± 40 μmol/min, P < .01). Urinary cyclic guanosine monophosphate (cGMP) excretion increased significantly and to a similar extent during ANP and ANP plus mannitol in all 12 patients, supporting the active biological responsiveness of renal ANP receptors. Unlike responders, nonresponders showed a significant decrease in arterial blood pressure and an increase in plasma renin activity during ANP plus mannitol, consistent with worsened arterial underfilling caused by ANP-induced vasodilation. Thus, the present results support the hypothesis that diminished distal tubular sodium delivery is a major factor contributing to ANP resistance in cirrhosis. (Hepatology 1995; 22:737-743.)
Hepatic distribution of E2 component of pyruvate dehydrogenase complex after transplantation
Authors: Neuberger James , Wallace Lorraine , Joplin Ruth , Hubscher Stefan
Published: March 2007

We have examined the distribution of the E2 component of pyruvate dehydrogenase complex in the liver of patients with native primary biliary cirrhosis (PBC), and after transplantation, using affinity-purified anti-E2 antibodies. In the posttransplantation group, we studied biopsy specimens from patients grafted for conditions other than PBC (n = 6) and those grafted for PBC with (n = 6) and without (n = 5) histological features suggestive of PBC recurrence. Features suggestive of PBC recurrence included portal tract granulomas, bile duct damage, ductopenia, and lymphoid aggregates. In the native liver from patients with PBC, there was increased staining of E2 on the biliary epithelial cells compared with hepatocytes, as previously described. However, in liver biopsy specimens from patients after transplantation, the pattern of staining of E2 was similar to that of normal, control liver in all three groups studied. These findings suggest that E2 overexpression on bile duct cells may not be important in the perpetuation of the bile duct damage in PBC, that expression in the allograft may be modified by immunosuppression, or that PBC does not recur in the allograft. (Hepatology 1995; 22:798-801.)
Long-term effect of nadolol or nadolol plus isosorbide-5-mononitrate on renal function and ascites formation in patients with cirrhosis
Authors: Merkel Carlo , Gatta Angelo , Donada Carlo , Enzo Edda , Marin Renato , Amodio Piero , Torboli Pierluigi , Angeli Paolo , Cavallarin Giorgio , Sebastianelli Giuliana , Susanna Sara , Mazzaro Cesare , Beltrame Pietro
Published: March 2007

The association beta-blockers plus nitrates has been reported to impair renal function and renal sodium handling, leading to increased risk of development of ascites, or worsening of a preexisting ascites, or increase in the requirements of diuretic agents. In 81 patients with cirrhosis and esophageal varices, participating in a multicenter controlled clinical trial of prophylaxis of variceal bleeding comparing nadolol (NAD) plus isosorbide-5-mononitrate (15M) with NAD alone, renal function, presence of ascites, and diuretic requirements were assessed at inclusion and after 6 months of follow-up. No significant variation in s-urea or s-creatinine was observed in either group. Three patients in the nadolol group and two in the NAD plus 15M developed ascites at 6 months (P = .70), and a need to increase diuretic regimen was observed in four and three patients, respectively (P = .76). Decrease in heart rate and in mean arterial pressure was similar in the two groups. There was a significant correlation between increase in s-creatinine and decrease in mean arterial pressure in the whole series (P = .015). Only in patients treated with the association was there a significant larger proportion of patients ascitic who became anascitic, than of patients anascitic who became ascitic (P = .03). In patients treated with the association, there was a significantly larger decrease in hepatic venous pressure gradient (P = .05). It is concluded that patients treated with the association NAD plus 15M are not at increased risk of developing renal dysfunction or worsening of ascites compared with patients treated with NAD alone. Therefore, the presence of ascites should not be considered a contraindication to the use of this association in patients with cirrhosis and portal hypertension. (Hepatology 1995; 22:808-813.)
Diclofenac-associated hepatotoxicity: Analysis of 180 cases reported to the food and drug administration as adverse reactions
Authors: Banks Alpha T. , Zimmerman Hyman J. , Ishak Kamal G. , Harter John G.
Published: March 2007

Diclofenac is a nonsteroidal anti-inflammatory drug approved in the United States in 1988 for the treatment of patients with osteoarthritis, rheumatoid arthritis, or ankylosing spondylitis. To characterize the clinical, biochemical, and histological features and possible mechanisms of hepatic injury associated with its use, a retrospective analysis was undertaken of 180 patients whose cases were reported to the Food and Drug Administration from November 1988 through June 1991, as having had possible adverse reactions to diclofenac. Of the reported 180 cases, 79% were female, 71% were 60 years of age or older, and 77% had osteoarthritis. Sixty-seven percent of the cases were detected by symptoms and the remainder by abnormal laboratory tests. Seventy-five percent of the symptomatic patients (90 of 120) were jaundiced. Seven of the 90 icteric patients died. The biochemical pattern of injury was hepatocellular or mixed hepatocellular in 66% of cases. Only 8% had a pattern of cholestatic injury. The remainder, with modestly increased values of both transaminases and alkaline phosphatase, were considered “indeterminate,” i.e., either mild hepatocellular or anicteric “cholestatic” injury. Sections of liver from 21 cases were available for study. Hepatic injury was apparent by 1 month after starting the drug in 24% by 3 months in 63%, and by 6 months in 85% of cases. The latent period in 12% was 6 to 12 months, whereas in 3% it was greater than 12 months. A combination of rash, fever, and eosinophilia, all hallmarks of immunological idiosyncrasy (hypersensitivity), was not reported in any case; additionally, the long latent period in most of the patients led to the inference that the mechanism is probably metabolic idiosyncrasy. The data suggest that diclofenac-related liver injury is particularly likely to involve osteoarthritic females, presenting with jaundice 1 to 6 months after starting diclofenac, with injury that is predominantly hepatocellular and presumably caused by metabolic idiosyncrasy. (Hepatology 1995; 22:820-827.)
N -ethyl-tauroursodeoxycholic acid, a novel deconjugation-resistant bile salt analogue: Effects of acute feeding in the rat
Authors: Angelico Mario , Mangiameli Andrea , Nistri Alessandra , Baiocchi Leonardo , Sofia Mara , Maina Mario , Di Martino Mario , Blasi Adriano
Published: March 2007

The purpose of this study was to investigate the physicochemical/biological properties and the effects of acute administration of N-ethyl-tauroursodeoxycholic acid in bile-fistula rats. In vitro determination of high-performance liquid chromatography mobility, octanol/water partitioning, cholesterol solubilizing capacity, and sensitivity to enzyme deconjugation by bacteria and cholylglycine-hydroxylase were performed. In vivo determination of the following was also performed: (1) maximum secretory rate (SRmax) and choleretic/secretory properties during intravenous (IV) administration; (2) site/extent of absorption, effects on bile flow, lipid secretion, and biotransformations after intraduodenal infusion. N-ethyl-tauroursodeoxycholate has a lipophilicity slightly higher than tauroursodeoxycholate, close to taurocholate, and similar cholesterol solubilizing capacity. Deconjugation of N-ethyl-tauroursodeoxycholate was 3.4 ± 2.1% after 72 hours, that of tauroursodeoxycholate was 100% after 24 hours. During IV infusion of 300 nmol/min/100 g, biliary secretion of N-ethyl-tauroursodeoxycholic and tauroursodeoxycholic acids averaged 185 ± 76 (standard deviation) nmol/min/100 g and 221 ± 77 nmol/min/100 g (not significant). Increasing infusion rates caused progressive enhancement of bile flow and bile salt secretion until the SRmax was reached (1,305 ± 240 nmol/min/100 g for N-ethyl-tauroursodeoxycholic acid and 3,240 nmol/min/100 g for tauroursodeoxycholate). The two bile salts were similarly choleretic. IV feeding of N-ethyl-tauroursodeoxycholic promoted a greater lipid secretion than tauroursodeoxycholate. After intraduodenal feeding of 800 μmol, 38.8 ± 14.0% and 43.4 ± 12.4% of the two bile salts were recovered in bile. No unconjugated bile salts nor unusual metabolites were detected. Excluding the distal 30 cm of the ileum, only 2.3 ± 0.7% of N-ethyl-tauroursodeoxycholic and 4.0 ± 2.3% of tauroursodeoxycholic acid was absorbed. N-ethyl-tauroursodeoxycholic is a new, hydrophilic, deconjugation-resistant, synthetic bile salt, which, in acute studies in the rat, behaves as the natural analogue with respect to intestinal and hepatobiliary properties. (Hepatology 1995; 22:887-895.)
Coexpression of liver-specific and growth-induced genes in perinatal and regenerating liver: Attainment and maintenance of the differentiated state during rapid proliferation
Authors: Haber Barbara , Naji Leyla , Chessman Drew , Taub Rebecca
Published: March 2007

The liver shows maximal cellular growth during fetal development and after partial hepatectomy. Exploring overlaps in gene expression patterns in these two types of hepatic growth may provide insight into common regulatory pathways. The expression of a large number of growth-induced and liver-specific genes induced in liver regeneration has been examined in the perinatal liver from several days prenatal to 4 weeks postnatal when the major growth phase of the liver ceases. As in liver regeneration, many growth-induced genes, such as PRL-1 and β-actin, are expressed at a high level throughout the temporal course of liver development and correlate with the proliferative state. The level of fetal liver expression of these genes is similar to peak expression found in the regenerating liver, suggesting that common pathways of transcriptional regulation exist in the two types of proliferation. A subset of liver-restricted immediate-early genes including, IGFBP-1, CL-6, and glucose-6-phosphatase (G6Pase) are induced in regenerating liver and may be important in maintaining hepatic metabolism during regeneration. In developing liver, these genes are expressed primarily in the perinatal period but, unlike the regenerating liver, are not coinduced. For instance, at birth, G6Pase is induced, whereas CL-6 is downregulated. In situ analyses confirm that a proliferation associated gene PRL-1 is expressed in multiple cell types throughout the developing liver, whereas the expression of liver-specific genes is confined to hepatocytes. Taken together, these findings imply that significant similarities and differences in transcriptional regulation and hormonal milieu exist in liver during regeneration and development. The increased expression of genes responsible for glucose homeostasis in proliferative states of the liver suggests that maintenance of differentiated function is a component of the hepatic growth response. (Hepatology 1995; 22:906-914.)
Downregulation of phorbol 12-myristate 13-acetate-induced tumor necrosis factor-alpha and interleukin-1β production and gene expression in human monocytic cells by human alpha-fetoprotein
Authors: Wang Wei , Alpert Elliot
Published: March 2007

We previously identified a specific receptor for alpha-fetoprotein (AFP) on human monocytes. Although AFP alters many immune cell functions, the effect of AFP on monocyte cytokine production is unknown. Because tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) are important cytokines in immunoregulation, we investigated whether AFP could modulate TNF-α and IL-1β production in U937, a human monocytic cell line. Our results showed that U937 cells secreted TNF-α and IL-1β in response to either phorbyl 12-myristate 13-acetate (PMA) or IFN-γ + LPS. In contrast, AFP significantly suppressed PMA-induced TNF-α and IL-1β production by U937 cells in a time and dose dependent fashion. Pretreatment of U937 cells with AFP resulted in maximal inhibition of PMA-stimulated TNF-α and IL-1β production by 58% and 67%, respectively. AFP also inhibited interferon-γ plus lipopolysaccharide (IFN-γ + LPS)-induced TNF-α and IL-1β production. Furthermore, Northern blot analysis showed that AFP suppressed PMA-mediated TNF-α and IL-1β messenger RNA (mRNA) expression. PMA-induced prostaglandin E2 (PGE2) production by U937 cells was enhanced by AFP. Pretreatment with indomethacin, a cyclooxygenase inhibitor, reversed AFP-inhibited TNF-α production by 78%. Thus, we conclude that AFP downregulates TNF-α and IL-1β production via a PGE2-dependent mechanism. (Hepatology 1995; 22:921-928.)
A fish oil diet minimizes hepatic reperfusion injury in the low-flow, reflow liver perfusion model
Authors: Zhong Zhi , Thurman Ronald G.
Published: March 2007

In this study, the effects of fish oil treatment on hepatic reperfusion injury in a low-flow, reflow perfusion model were investigated. Rats were fed powdered diets containing either 5% corn oil or 5% encapsulated fish oil for 13 to 15 days. Average daily food intake in both control and fish oil groups was about 20 g per rat, and weight gain averaged 9 g/rat/d. Livers were perfused at flow rates around 1 mL/g/min for 75 minutes, which caused cells in pericentral regions to become anoxic because of insufficient delivery of oxygen. When normal flow rates (about 4 mL/g/min) were restored for 40 minutes, reperfusion injury occurred. Upon reflow, lactate dehydrogenase (LDH) release increased from basal levels around 1 to 50 IU/g/h hi livers from control rats, whereas fish oil treatment minimized values to 16 IU/g/h. Rates of bile production reached 23 μL/g/h during reperfusion in livers from controls and 38 μL/g/h in the fish oil-treated group. Oxygen uptake was about 110 μmol/g/h during the reperfusion period in livers from both groups. Malondialdehyde (MDA), an end product of lipid peroxidation, was released into the effluent perfusate at rates around 80 nmol/g/h during reperfusion in controls, and values were not affected by fish oil treatment. Portal pressure, an indicator of hepatic microcirculation, increased from basal levels of 3 to 10.5 cm H2O during reperfusion in controls, but was only elevated to 8.3 cm H2O in the fish oil-treated group. In addition, Trypan Blue distribution time, another indicator of hepatic microcirculation, was reduced significantly by 44% by fish oil treatment. It is concluded that fish oil minimized hepatic reperfusion injury, most likely by reducing the production of vasoconstrictive eicosanoids from arachidonic acid, thereby improving the hepatic microcirculation, which prevents cell death on reperfusion. (Hepatology 1995;22:929-935.)
Processing of endoplasmic reticulum luminal antigens associated with halothane hepatitis in rat hepatocytes
Authors: Amouzadeh Hamid R. , Pohl Lance R.
Published: March 2007

In this study we have investigated the mechanism of the processing of trifluoroacetylated liver microsomal protein antigens associated with halothane hepatitis to learn how the immune system might come in contact with these proteins to form antibodies directed against them. Rats were treated with halothane and parenchymal (PC) and non-parenchymal cells (NPC) were isolated 16 hours later. Immunoblotting of the cell lysates with antisera directed against the trifluoroacetyl hapten showed the presence of high levels of trifluoroacetylated proteins in parenchymal cells, whereas none of these proteins were detected in endothelial or Kupffer cells that were isolated by centrifugal elutriation. The halflives of 100-, 82-, 80-, 63-, 59-, 58-, and 57-kd trifluoroacetylated and native carrier proteins of the trifluoroacetyl hapten in cultures of rat primary parenchymal cells were approximately 1 day. The turnovers of all of these trifluoroacetylated proteins, except for that of the trifluoroacetylated 100-kd protein, were inhibited by treatment of the cells with ammonium chloride, leupeptin, 4-(2-aminoethyl)-benzenesulfonyl fluoride, or 3-methyladenine (3-MA). These results indicate that, in liver, the major source of the formation of trifluoroacetylated antigens associated with halothane hepatitis is the parenchymal cells. It appears that most of the trifluoroacetylated antigens and possibly the native carrier protein of the trifluoroacetyl haptens are transferred from the endoplasmic reticulum (ER) to an acidic compartment of PCs, where they are enzymatically degraded. The processing of the trifluoroacetylated proteins by this pathway may be a protective mechanism that prevents these covalently altered proteins from inducing an antibody response in most patients who are administered halothane. (Hepatology 1995; 22:936-943.)
Expression of glycoconjugates during intrahepatic bile duct development in the rat: An immunohistochemical and lectin-histochemical study
Authors: Sanzen Takahiro , Yoshida Kazuharu , Sasaki Motoko , Terada Tadashi , Nakanuma Yasuni
Published: March 2007

We investigated the expression of carbohydrate residues on the developing intrahepatic bile ducts of rats. At 17 days of gestation, immature biliary cells around the portal vein close to the hepatic hilum assumed one of the following forms: slitlike lumen, incomplete, or complete bile ductule-like structures. These immature biliary elements then rapidly spread throughout the liver along with development. At birth, a few mature interlobular bile ducts became visible in the portal tracts. The cytoplasm of immature biliary cells stained weakly for concanavalin A, Erythrina crista galli agglutinin, and Limax flavus agglutinin, whereas the luminal surface of immature biliary cells at 17 days of gestation was positive for lectins, similar to those that are expressed on the luminal surface of the mature bile ducts, including concanavalin A, succinyl wheat germ agglutinin, Vicia villosa agglutinin, soybean agglutinin, peanut agglutinin, Erythrina crista galli agglutinin, and Limax flavus agglutinin. As development progressed, the number of lectins binding to the cytoplasm of biliary cells gradually increased, and lectin bindings to the luminal surface of biliary cells gradually became intense. Immature biliary epithelial cells of three structures expressed similar carbohydrate residues in their cytoplasm and luminal surfaces. This study suggested that the profile of carbohydrate residues on the biliary epithelium changes with development. Therefore, this profile could be a useful tool with which to evaluate the development of the biliary tree as well as associated disorders. (Hepatology 1995; 22:944-951.)
Characteristic difference of hepatocellular carcinoma between hepatitis B- and C- viral infection in Japan
Authors: Shiratori Yasushi , Shiina Shuichiro , Imamura Masatoshi , Kato Naoya , Kanai Fumihiko , Okudaira Takahito , Teratani Takuma , Tohgo Gohichi , Toda Nobuo , Ohashi Makoto , Ogura Keiji , Niwa Yasuro , Kawabe Takao , Omata Masao
Published: March 2007

Characteristics of 205 consecutive patients with hepatocellular carcinoma (HCC) admitted during 1990 to 1993 have been analyzed from the standpoint of hepatitis viral infection in Japan. Among 205 HCC patients, 71% of the patients showed positivity for hepatic C virus (HCV) antibody alone, 13% showed positivity both for HCV and HBV (HCV/HBV) antibody, 11% demonstrated HBsAg alone, and negativity of both HCV and HBV antibody in 4% only. Positivity to both HCV antibody and HBsAg was demonstrated in 1% only. Mean detection age of HCVAb-positive HCC as well as both HCV/HBV antibody-positive HCC was 62 ± 7 years, in contrast to 52 ± 13 years in HCC with HBsAg (P < .05). Male-to-female ratio among HCVAb-positive HCC was 3.3:1, in contrast to 5.5:1 among the HCV/HBVAb-positive HCC and 7:1 among HBsAg-positive HCC, but there was no significant difference in the gender distribution between these groups. More than 60% of HCVAb-positive HCC were classified into the stage of Child B and C, whereas 65% of HBsAg-positive HCC was at the stage of Child A. The severity of liver disease was confirmed by liver histology, indicating that more than 70% of the HCVAb-positive HCC and the HCV/HBVAb-positive HCC showed cirrhosis, in contrast to 50% among the HBsAg-positive HCC. Three-year survival rate of HCV Ab-positive HCC and HBV/HCVAb-positive HCC was 68% and 56%, respectively, in contrast to 47% in HBsAg-positive HCC. HCC was found at early stage among the patients receiving periodic medical checkups beforehand, and the prognosis of these HCC patients was significantly better than those who did not receive checkups. From these results, HCV-related HCC occupied over 80% of total HCC in Japan, which are characterized by older age and more severe cirrhosis, as compared with HBsAg-positive HCC. Prognosis of HCV Ab-positive HCC was not significantly better than HBsAg-positive HCC, but the periodical screening naturally improves prognosis because the cases are found usually much earlier (lead time bias) and mostly belong to slow progression type (length bias). (Hepatology 1995; 22:1027-1033.).
The influence of a maternal chronic hepatitis B virus infection on the repertoire of transcribed T-cell receptor beta chain variable region genes in human cord blood
Authors: Abbott William G. H. , Geursen Arie , Fraser John D. , Marbrook John , Skinner Margot A. , Tan Paul L. J.
Published: March 2007

We used an anchor polymerase chain reaction method to compare the repertoires of transcribed T-cell receptor β chain variable region (Vβ) genes in cord blood T cells from neonates of hepatitis B surface antigen (HBsAg) positive (n = 40) and HBsAg negative (n = 40) women. Fifteen of the HBsAg positive women were hepatitis B e antigen (HBeAg) positive, and 25 were HBeAg negative. The percentage of Vβ.4 transcripts was lower in cord blood T cells from neonates of HBsAg-positive relative to HBsAg-negative women (9.7% ± 0.5% vs. 12.7% ± 0.6%, P = .002). The percent of Vβ5.1 transcripts was higher in cord blood T cells from neonates of HBeAg-positive relative to HBeAg-negative women (9.3% ± 0.7% vs. 7.0% ± 0.3%, P < .001). There were no correlations between neonatal maturity at birth and Vβ repertoire. In summary, a maternal chronic hepatitis B virus (HBV) infection is associated with changes in the repertoire of transcribed T-cell receptor genes in neonatal cord blood T cells. It is possible that the T-cell response to the HBV is associated with a limited repertoire of Vβ genes. The mechanism of vertical chronic HBV infection in human neonates may involve changes in the T-cell response to the virus that are induced in utero. (Hepatology 1995; 22:1034-1039.).
Induction of T-helper cell response to hepatitis B core antigen in chronic hepatitis B: A major factor in activation of the host immune response to the hepatitis B virus
Authors: Marinos George , Torre Francesco , Chokshi Shilpa , Hussain Munther , Clarke Berwin E. , Rowlands David J. , Eddleston Adrian L. W. F. , Naoumov Nikolai V. , Williams Roger
Published: March 2007

The T helper (Th) cell response to hepatitis B core antigen (HBcAg) was analyzed in 76 chronic hepatitis B virus (HBV) carriers with varying degrees of hepatic inflammation and HBV replication. Fifty-five patients had active viral replication, 28 with minimal histological changes and normal alanine transaminase (ALT) and 27 with active hepatic inflammation and elevated ALT. The remaining 21 chronic hepatitis B surface antigen (HBsAg) carriers had undetectable HBV replication, minimal histological activity, and normal ALT. In addition, 34 chronic HBV carriers were studied prospectively during treatment with α-interferon. The HBcAg-specific Th cell response was evaluated by a proliferative assay using 3H-thymidine uptake and γ-interferon production by peripheral blood mononuclear cells. The proliferative response and γ-interferon production of patients with active hepatic inflammation were significantly higher than in patients with minimal histological changes and in controls. In the longitudinal analysis during α-interferon treatment, 22 of 34 patients sustained an ALT flare accompanied by a parallel, significant Th cell response, which preceded or coincided with the ALT flare. The elevation in the Th cell response and the ALT flare were followed by a significant rise in the serum immunoglobulin (Ig) M anti-HBc index. Ten of twenty-two patients with an enhanced Th cell response and an ALT flare seroconverted after α-interferon treatment. The Th cell activity in the 10 responders rapidly subsided after hepatitis B e antigen (HBeAg) to anti-HBe seroconversion, whereas in the 12 nonresponders it remained elevated. This study demonstrates that patients with chronic hepatitis B (CAHB) have a detectable and a significant Th cell response to HBcAg, which is likely to be involved in augmenting the immune-mediated hepatocellular damage and in the activation of HBV-specific humoral immune reaction. Thus, loss of Th cell nonresponsiveness to HBcAg is an important factor in enhancing the effector immune responses to HBV in chronic hepatitis B. (Hepatology 1995; 22:1040-1049.).
Interferon gamma production by peripheral blood lymphocytes to hepatitis C virus core protein in chronic hepatitis C infection
Authors: Iwata Kazuo , Wakita Takaji , Okumura Akihiko , Yoshioka Kentaro , Takayanagi Masahiro , Wands Jack R. , Kakumu Shinichi
Published: March 2007

Evidence suggests that cellular immunity to hepatitis C virus (HCV) core protein may be important in the pathogenesis of viral infection. Therefore, interferon gamma (IFN-γ) production by peripheral blood mononuclear cells (PBMC) derived from patients with chronic HCV infection (genotype 1b) was examined. The cellular immune response was evaluated with a recombinant HCV core fusion protein derived from a patient with genotype 1b. To identify the immunodominant epitopes, IFN-γ production in responders was also assessed with a panel of nine synthetic peptides that covered the entire core region. It was found that mononuclear cells from 24 (52%) of 46 patients with chronic liver disease responded to the core protein; asymptomatic HCV carriers demonstrated a lower response rate (14%, P < .05). More important, individuals who had received IFN-α treatment and went into clinical and virological remission had a higher response rate (75%, P < .05) compared with those with ongoing hepatitis whose treatment failed (31%). Of 25 patients whose mononuclear cells responded to HCV core protein, 18 had a significant response to one or more peptides; 12 patients reacted to a peptide mixture containing hydrophilic sequences. The core peptide amino acid sequence 141 to 160 was recognized in 9 patients. Interestingly, 7 of 8 patients bearing HLA DR 4 and w53 haplotypes recognized the peptide sequence 141 to 160. Thus, IFN-γ production of the mononuclear cell response appeared to be HLA DR restricted, and the responding cells were identified as CD4+ T cells. This study suggests the presence of immunodominant T cell epitopes within the HCV core protein in association with HLA DR phenotypes in patients with HCV-associated liver disease. (Hepatology 1995;22:1057-1064.).
An open trial of interferon alfa recombinant for hepatitis C after liver transplantation: Antiviral effects and risk of rejection
Authors: Féray Cyrille , Samuel Didier , Gigou Michèle , Paradis Valérie , David Marie Francoise , Lemonnier Catherine , Reynès Michel , Bismuth Henri
Published: March 2007

The aim of this open trial was to assess the efficacy and the safety of interferon (IFN) alfa therapy in liver transplant recipients with chronic active hepatitis caused by hepatitis C virus. In July 1991, among 447 liver recipients regularly observed at our institution, 46 had developed HCV-related chronic active hepatitis defined by piece meal necrosis. Fourteen of these 46 patients received IFN alfa 3 mIU three times weekly for a planned duration of 6 months and were compared to the 32 untreated patients. Genotyping and quantification of viremia were performed using type-specific amplification and branched DNA assay. Histological follow-up was available in all patients and routinely before and after IFN therapy. Treated and untreated patients did not differ regarding gender, age, length of follow-up, maximum histological score, and genotypes (41 of 46 were of type 1b). Induction of chronic rejection was observed in 5 of 14 treated patients leading to retransplantation in 3. In contrast, chronic rejection occurred in 1 of 32 untreated patients (P < .005) during the posttransplantation follow-up. Among the 9 treated patients without rejection, a decrease of transaminases or of HCV RNA levels of more than 50% were observed in 8 and 4, respectively; 2 patients had a complete response, and 1 did not relapse after discontinuation of IFN. Histological improvement occurred in 2 of the treated patients and in none of the untreated patients. IFN therapy in liver transplant recipients has poor antiviral effect and can induce chronic rejection. Its use in this setting should be cautious. (Hepatology 1995; 22:1084-1089.).
Utility of hepatitis C virus RNA determinations in hepatic tissue as an end point for interferon treatment of chronic hepatitis C
Authors: Gurakar Ahmet , Fagiuoli Stefano , Faruki Hawazin , De Maria Nicola , Balkan Mujdat , Van Thiel David H. , Friedlander Lois
Published: March 2007

A total of 41 patients with chronic hepatitis C virus (HCV) defined as abnormal liver injury test results for 6 months or more and HCV RNA positivity in plasma were studied to determine if the liver might not be the only focus of HCV infection in individuals treated with interferon alfa (IFN-α). All patients were examined for the presence of confounding liver disease and tested negatively for such findings. All tested positively for HCV RNA and had an abnormal hepatic histology. All were treated with IFN for 6 months at a dosage of 5 million units daily. After 6 months of therapy, 29 (71%) had normal alanine transaminase (ALT) values, and 25 (61%) tested negatively for HCV RNA. After 6 months of follow-up, without IFN therapy, 17 (41%) still had normal ALT values, and 16 (39%) still tested negatively for HCV RNA in serum. Patients who continued to test negatively for HCV RNA in serum after 6 months of follow-up also tested negatively for HCV RNA in the liver at the end of IFN therapy. Only 2 subjects who tested negatively for HCV RNA in the liver at the end of treatment relapsed after discontinuing IFN therapy. In contrast, patients who tested positively for HCV RNA in the liver after 6 months of therapy relapsed and tested positively for HCV RNA in serum during the 6 months of follow-up. These results suggest that (1) a nonhepatic site of HCV infection may exist; (2) this putative extrahepatic site appears to be less responsive to IFN therapy than in the liver; and (3) this unknown extrahepatic site of infection may be the source of HCV reactivation in cases that relapse during IFN follow-up periods. (Hepatology 1995; 22:1109-1112.).
Donor selection limits use of living-related liver transplantation
Authors: Renz John F. , Mudge Chris L. , Heyman Melvin B. , Tomlanovich Steve , Kingsford Ralph P. , Moore Barbara J. , Snyder John D. , Perr Hilary A. , Paschal Amie L. , Roberts John P. , Ascher Nancy L. , Emond Jean C.
Published: March 2007

The purpose of this investigation was to assess the applicability of living-related liver transplantation in an established regional transplant program by determining the frequency of acceptable living donors from an unselected population of pediatric transplant candidates and identify specific factors limiting application of this technique. During the period May 1992 to May 1994, all children accepted as transplant candidates at the University of California-San Francisco were evaluated for potential living-related liver transplantation. Indications for transplantation and patient demographics represented the spectrum anticipated at a regional center. Donor evaluation was performed using a three-phase evaluation process we have previously reported. Retrospective analysis identified 75 potential donors for 38 pediatric candidates (age range, 17 days to 14.5 years; mean, 5.1 years). Twenty-three percent of potential donors declined evaluation. Of the 75 potential donors, only 10 (13%) were found to be acceptable for donation. The leading causes for donor declination were significant medical history (23%), ABO blood group incompatibility (23%), and psychosocial history (20%). Of the 38 recipient candidates, 9 (23%) were offered living-donor transplantation. Five patients have received living-donor transplantation, and 4 patients await the procedure when medical indications exist. Seventy-seven percent of recipient candidates received or are awaiting cadaveric transplantation. These results suggest that current donor criteria markedly limit the application of living-related liver transplantation. (Hepatology 1995; 22:1122-1126.).
Intrahepatic mast cells in chronic liver diseases
Authors: Farrell Desmond J. , Hines Julie E. , Walls Andrew F. , Kelly Peter J. , Bennett Mark K. , Burt Alastair D.
Published: March 2007

Mast cells are known to be present in human liver but their distribution and density in normal livers and in chronic liver diseases have not previously been examined. In this study, we quantified mast cell numbers and examined their distribution in percutaneous biopsy specimens from normal livers (n = 8) and in two chronic progressive liver diseases: primary biliary cirrhosis (PBC) (n = 40) and alcoholic liver disease (n = 33). We compared differences in mast cell density between these two forms of chronic liver disease because it had been suggested that mast cells may play a role in the development of liver fibrosis, particularly in patients with chronic cholestatic liver disease who frequently have increased plasma histamine levels. Mast cells were identified by immunohistochemistry using a specific monoclonal antibody (AA1) raised against mast cell tryptase after an initial study showed this to be more sensitive for the detection of mast cells than the conventional histochemical stain, toluidine blue. Our results showed that small numbers of mast cells (3.9 ± 3.3/mm2) are present within the portal tracts and sinusoids of normal livers. In progressive chronic liver disease, increased numbers of mast cells were present, which correlated with the increasing amounts of liver fibrosis present. We found significantly more mast cells in the PBC group compared with the alcoholic group for a given amount of fibrosis. Our findings suggest that mast cells and their mediators may play a role in liver fibrogenesis. (Hepatology 1995; 22:1175-1181.).
Sequential acetaldehyde production, lipid peroxidation, and fibrogenesis in micropig model of alcohol-induced liver disease
Authors: Nlemelä Onni , Parkkila Seppo , Ylä-herttuala Seppo , Villanueva Jesus , Ruebner Boris , Halsted Charles H.
Published: March 2007

The pathogenesis of alcohol-induced liver disease involves the adverse effects of ethanol metabolites and oxidative tissue injury. Previous studies indicated that covalent protein adducts with reactive aldehydes may be formed in alcohol consumers. To study the role of such protein adducts in the development of liver injury, we examined the sequential appearances of adducts of the ethanol metabolite acetaldehyde (AA) and of two products of lipid peroxidation, malondialdehyde (MDA) and 4-hydroxynonenol (HNE), in ethanol-fed micropigs. Immunohistochemical stainings using specific antibodies that recognize epitopes of each adduct were performed from liver biopsy specimens obtained at 1, 5, and 12 months from micropigs fed either control diet (n = 5) or ethanol-containing diets (n = 5). After 1 month on the ethanol diet, AA and MDA adducts were observed primarily in the perivenous regions co-localizing with each other and coinciding with increased concentrations of serum aminotransferase markers of liver injury. HNE adducts were usually less intense and more diffuse, and were also seen in some biopsy specimens from control animals. Although the most intense staining reactions at 5 months remained in zone 3, a more widespread distribution was usually seen together with increased evidence of steatonecrosis and focal inflammation. In terminal biopsies at 12 months, perivenous fibrosis was present in three of five biopsy specimens. More extensive pericentral and intralobular fibrosis was noted in one micropig fed ethanol for 21 months. These studies demonstrate that covalent adducts of proteins with reactive aldehydes are formed in early phases of alcohol-induced liver disease. Perivenous adducts of the AA product of ethanol metabolism and the MDA product of lipid peroxidation appear to precede necrosis and fibrosis. (Hepatology 1995; 22:1208-1214.).
Elevated mitochondrial gene expression during rat liver regeneration after portal vein ligation
Authors: Shimizu Yasuhiro , Suzuki Hiroshi , Nimura Yuji , Onoue Shigemi , Nagino Masato , Tanaka Masashi , Ozawa Takayuki
Published: March 2007

We explored the molecular basis of mitochondrial energy production during rat liver regeneration after portal vein ligation. Ligation of the left branch of the portal vein induces an increase in the weight of the nonligated lobe, counterbalancing the reduced weight of the ligated lobe. Using this model, we investigated changes in mitochondrial DNA-binding proteins, mitochondrial DNA, and mitochondrial messenger RNA (mRNA) in rat hepatocytes of the nonligated lobes. The amount of mitochondrial DNA-binding protein increased maximally (200% to 300% of the preoperative level) at 12 hours after the operation, before an increase (390%) in mitochondrial DNA content at 24 hours, and parallel to an increase (240%) in mitochondrial mRNA levels at 12 hours. These results suggest that the energy supply for liver regeneration is achieved through enhancement of mitochondrial DNA replication as well as transcription, in which the mitochondrial DNA-binding proteins probably play regulatory roles. We also found that in the nonligated lobes, mRNA levels of hepatocyte growth factor increased to a detectable level only 12 hours after the operation. These molecular biochemical data help explain why preoperative portal vein embolization, which is a modification of portal vein branch ligation, is an effective method to prevent posthepatectomy liver failure. (Hepatology 1995; 22:1222-1229.).