bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiple-dose pharmacokinetics of rufloxacin in patients with cirrhosis
Authors: Triger David R. , Granai Fernando , Woodcock Jocelyn , Wise Richard , Imbimbo Bruno P.
Published: March 2007

The multiple-dose pharmacokinetics of rufloxacin were investigated in 13 patients with biopsy-proven cirrhosis and in 5 healthy controls. Rufloxacin was administered once a day for 5 consecutive days, starting with a loading dose of 400 mg on day 1 and 200 mg on the subsequent days. Plasma and urinary drug concentrations were determined by high-performance liquid chromatography and a microbiological assay. A one-compartment model applied to the high-performance liquid chromatography data was used to calculate the pharmacokinetic parameters of rufloxacin. In the controls rufloxacin had a low plasma clearance (41 ± 4 ml/min, mean ± S.E.M.), a long half-life (30.1 ± 3.9 hr), a large area under the plasma concentration vs. time curve (171 ± 18 μg · hr/ml) and a low renal clearance (18 ± 2 ml/min). No appreciable differences were observed in the pharmacokinetic parameters between patients with various degrees of liverfunction impairment (modified Child-Pugh score ranging from 5 to 13). In these patients plasma clearance was slightly reduced (−32%), but this decrease was caused by a marked reduction in renal clearance (−65%) rather than nonrenal clearance, which remained unchanged (22 ml/min in cirrhotic patients vs. 23 ml/min in controls). A significant (p < 0.01) correlation was found between creatinine clearance and both rufloxacin renal clearance (r = 0.769) and rufloxacin plasma clearance (r = 0.681). The elimination half-life and the area under the plasma concentration vs. time curve were moderately increased in cirrhotic patients (+33% and +26%, respectively). These findings indicate that factors related to declining renal function, rather than liver impairment itself, may be the prime cause in determining alterations in the pharmacokinetic parameters in cirrhotic patients. When these patients have normal renal function, no dose adjustment is needed during oral rufloxacin treatment. (HEPATOLOGY 1993;18:847-852).
Disordered hemostasis in extrahepatic portal hypertension
Authors: Robson Simon C. , Kahn Delawir , Kruskal J. , Bird Arthur R. , Kirsch Ralph E.
Published: March 2007

To assess the contribution of naturally occurring portal-systemic shunts to the coagulopathy of patients with liver disease, we studied laboratory parameters of hemostasis in 20 adult patients with extrahepatic portal hypertension, secondary to portal vein thrombosis, that had resulted in variceal bleeding. All extrahepatic portal hypertension patients had normal liver function and histological appearance. None had any evidence of preexisting coagulation disorders, and none had bled or undergone sclerotherapy in the 6 mo before study. Age- and gender-matched groups of 20 healthy individuals and 20 stable patients with cirrhosis and portal hypertension who had a history of variceal bleeding served as controls. Both patient groups had thrombocytopenia consistent with hypersplenism and portal hypertension. Prothrombin international normalized ratio (extrahepatic portal hypertension, 1.3 ± 0.12; cirrhosis, 1.7 ± 0.2; control, 1.02 ± 0.06; p < 0.05) and partial thromboplastin time ratios (extrahepatic portal hypertension, 1.12 ± 0.1; cirrhosis, 1.26 ± 0.2; controls, 1.01 ± 0.03; p < 0.05) were significantly prolonged in both patient groups. Extrahepatic portal hypertension and cirrhotic patient groups had significantly increased levels of serum total fibrin(ogen)-related antigen (extrahepatic portal hypertension, 818 ± 150 ng/ml; cirrhosis, 454 ± 52 ng/ml; controls, 124 ± 7.3 ng/ml; p < 0.05), fibrin monomer (extrahepatic portal hypertension, 168.8 ± 16.9 ng/ml; cirrhosis, 115.6 ± 11.1 ng/ml; controls, 19.7 ± 0.4 ng/ml; p < 0.05) and D-dimer (extrahepatic portal hypertension, 118 ± 9.6 ng/ml; cirrhosis, 129 ± 10 ng/ml; controls, 53.2 ± 1.6 ng/ml; p < 0.05). Extrahepatic portal hypertension patients had higher total fibrinogen levels (4.15 ± 1.4 gm/L) than did controls (3.35 ± 0.21 gm/L; p < 0.05). Patients with extrahepatic portal hypertension (tissue plasminogen activator, 22.1 ± 7; plasminogen activator inhibitor type 1, 51 ± 26) and cirrhosis (tissue plasminogen activator, 23.8 ± 15; plasminogen activator inhibitor type 1, 65 ± 30) had significantly higher concentrations of circulating tissue plasminogen activator and plasminogen activator inhibitor type I than did controls (tissue plasminogen activator, 5.8 ± 1.1 ng/ml; plasminogen activator inhibitor type 1, 5.2 ± 1.8 ng/ml; p < 0.05 for both comparisons). Analysis of specific coagulation factors revealed reduced levels of factors V, VII and IX, with increases of factor VIII levels in both patient groups suggestive of compensated mild, disseminated intravascular coagulation. Our data suggest that portal systemic shunting alters parameters of hemostasis, even in the apparent absence of liver disease, and raise the possibility that such shunting is responsible, at least in part, for the disordered coagulation and fibrinolysis seen in patients with chronic liver disease. (HEPATOLOGY 1993;18:853-857).
Halothane hepatitis patients have serum antibodies that react with protein disulfide isomerase
Authors: Martin Jackie L. , Kenna Gerald J. , Martin Brian M. , Thomassen David , Reed George F. , Pohl Lance R.
Published: March 2007

Clinical and laboratory evidence suggests that the fulminant liver failure sometimes associated with the inhalation anesthetic halothane may be an immunemediated toxicity. Most importantly, the vast majority of patients with a clinical diagnosis of halothane hepatitis have serum antibodies, which react with one or more specific liver microsomal proteins that have been covalently altered by the trifluoroacetyl chloride metabolite of halothane. The serum antibodies are specific to halothane hepatitis patients and are not seen in sera of patients with other types of liver pathology. In this study, a 57-kD trifluoroacetylated liver microsomal neoantigen associated with halothane hepatitis and native 57-kD protein were purified from liver microsomes of halothane-treated and -untreated rats, respectively. When the purified trifluoroacetylated 57-kD and native 57-kD proteins were used as test antigens in an enzyme-linked immunosorbent assay, serum antibodies from halothane hepatitis patients (n = 40) reacted with both of these proteins to a significantly greater extent than did serum antibodies from control patients (n = 32). On the basis of its apparent monomeric molecular mass, isoelectric point and NH2-terminal amino acid and tryptic peptide sequences, the 57-kD protein has been identified as rat liver protein disulfide isomerase. Antibodies raised against rat liver protein disulfide isomerase also reacted with a protein of approximately 58-kD in human liver microsomes. The results of this investigation suggest that trifluoroacetylated protein disulfide isomerase is one of the immunogens associated with halothane hepatitis. In certain patients it might lead either to specific antibodies or, possibly, to specific T cells, which could be responsible for halothane hepatitis. (HEPATOLOGY 1993;18:858-863).
Zonal distribution of protein-acetaldehyde adducts in the liver of rats fed alcohol for long periods
Authors: Lin Renee C. , Zhou Feng C. , Fillenwarth Michael J. , Lumeng Lawrence
Published: March 2007

Acetaldehyde, a highly reactive intermediate of alcohol metabolism, has been shown to form adducts with liver proteins in rats fed alcohol for long periods. In this report, the zonal distribution of liver proteinacetaldehyde adducts that formed in vivo was studied by means of histoimmunostaining. Rats were pair-fed alcohol-containing and alcohol-free AIN'76 liquid diets for 2 or 11 wk before they were killed and subjected to whole body perfusion with paraformaldehyde. Each liver was cut into 60-μm-thick slices. Slices were first treated with 10% hydrogen peroxide to eliminate endogenous peroxidase activity. They were then incubated sequentially with rabbit antihemocyanin-acetaldehyde adduct, goat antirabbit serum IgG and rabbit peroxidase-antiperoxidase complex. The liver slices were stained with diaminobenzidine and counterstained with methylgreen. In the livers of rats fed alcohol for 2 wk, peroxidase activity was evident in the perivenous zone but not the periportal zone. No staining was obtained when the primary antibody had been preabsorbed with immobilized hemocyanin-acetaldehyde adduct or if the liver slices were incubated with the unimmunized rabbit IgG. Slight staining of the perivenous zone was seen in the livers of control rats, presumably because of minimal protein-acetaldehyde adduct formation emanating from endogenous acetaldehyde. When rats were fed alcohol for longer periods (e.g., 11 wk), protein-acetaldehyde adducts were still seen predominantly in the perivenous zone, but the distribution pattern was more diffuse than that observed in the livers of rats fed alcohol for only 2 wk. More liver cells produced protein-acetaldehyde adducts when rats were fed the alcohol-containing diet supplemented with cyanamide. However, these protein-acetaldehyde adduct-positive cells were still found mainly in the perivenous area. The zonation in the formation of protein-acetaldehyde adducts in the liver may in part explain the preferential damage of perivenous hepatocytes induced by long-term alcohol consumption. (HEPATOLOGY 1993;18:864-869).
Microtubule-dependent transport of bile salts through hepatocytes: Cholic vs. taurocholic acid
Authors: Crawford James M. , Crawford Aleta R. , Strahs Deborah C. J.
Published: March 2007

Studies with taurine-conjugated bile salts have demonstrated two pathways for hepatocellular delivery of bile salts to bile: a cytosolic, microtubule-independent pathway and a membrane-based, microtubule-dependent pathway. However, a significant portion of circulating bile salts may be unconjugated. To determine whether free bile salts utilize similar pathways, we examined the effect of colchicine on the biliary excretion of intravenously administered cholic acid and taurocholate in intact rats. Basal rats were pretreated with low-dose colchicine or its inactive isomer, lumicolchicine, 1 hr before placement of intravenous and biliary cannulas and 2.75 hr before intravenous injection of [14C]cholic acid and [3H]taurocholate. Superfused rats were prepared as above but with intravenous infusion of taurocholate at 200 nmol/min 100 gm beginning 0.75 hr before [14C]cholic acid/[3H]taurocholate injection. Depleted/reinfused rats were subjected to biliary diversion for 20 hr before coichicine or lumicolchicine pretreatment, infusion of taurocholate and [14C]cholic acid/[3H]taurocholate injection. In each group, biliary excretion of [14C]taurocholate and [3H]taurocholate was inhibited equally by colchicine; for peak excretion rates the respective inhibition values were 33% and 35% in basal rats, 63% and 65% in superfused rats, and 74% and 76% in depleted/reinfused rats. Biliary excretion of [14C]taurocholate occurred consistently later than excretion of [3H]taurocholate, and maximal rates of excretion were reduced. In contrast, plasma uptake rates of [14C]cholic acid and [3H]taurocholate were essentially the same in depleted/reinfused rats. Deconvolution analysis of [14C]taurocholate vs. [3H]taurocholate biliary excretion curves revealed no significant differences among experimental groups. We conclude that conversion of [14C]cholic acid to [14C]taurocholate slightly retards its biliary excretion and diminishes its peak excretion rate compared with exogenous [3H]taurocholate. However, there appear to be no other differences in hepatocellular processing of the unconjugated and conjugated bile salts: taurocholate generated in the cytosol or taken up from plasma enters the same intracellular pathways for biliary excretion under different conditions. (HEPATOLOGY 1993;18:903-911).
Origin and differentiation of hepatic natural killer cells (pit cells)
Authors: Vanderkerken Karin , Bouwens Luc , de Neve Wilfried , van den Berg Kit , Baekeland Marijke , Delens Nadia , Wisse Eddie
Published: March 2007

Liver sinusoids contain a population of large granular lymphocytes or natural killer cells, originally termed pit cells. After isolation and purification, these cells were separated into a low-density and a high-density fraction. The liver low-density fraction differs significantly in morphology and function from cells of the blood, whereas the liver high-density fraction shows intermediate properties. In this study we demonstrate that this morphological and functional heterogeneity is based on subsequent steps of differentiation of the large granular lymphocytes within the liver. When cell proliferation was suppressed by sublethal total body irradiation, the life span of the hepatic large granular lymphocytes could be determined: high-density and low-density populations were totally depleted within 1 and 2 wk after irradiation, respectively. By using intravenous asialo-GM1 antiserum to deplete animals of asialo-GM1-positive cells, we found that the depletion of the asialo-GM1-positive cells preceded the depletion of asialo-GM1-negative hepatic low-density large granular lymphocytes by approximately 1 wk. Direct evidence that the asialo-GM1-positive high-density large granular lymphocytes are precursors of the low-density large granular lymphocytes was given by adoptive transfer experiments with fluorescent-labeled high-density cells. Three days after their injection, labeled large granular lymphocytes were found in the hepatic low-density fraction of the recipient rat, and these cells had developed morphological characteristics of low-density large granular lymphocytes. It is concluded therefore that marginating blood large granular lymphocytes differentiate through high-density large granular lymphocytes into the typical liver-specific low-density large granular lymphocytes or pit cells. (HEPATOLOGY 1993;18:919-925).
Centrilobular distribution of acetaldehyde and collagen in the ethanol-fed micropig
Authors: Halsted Charles H. , Villanueva Jesús , Chandler Carol J. , Ruebner Boris , Munn Robert J. , Parkkila Seppo , Niemelä Onni
Published: March 2007

We established a new animal model of alcoholic liver disease in the micropig, a species that consumes ethanol voluntarily in the diet. Ten micropigs were pair-fed diets containing 40% of calories as ethanol or cornstarch with identical amounts of fat, protein and micronutrients for 12 mo. Liver histopathology in the ethanol-fed pigs included steatonecrosis in all five and interstitial and perivenous fibrosis in three. Electron microscopy showed Ito-cell transformation with perisinusoidal collagen accumulation. Acetaldehyde adducts were found by immunofluorescence in the centrilobular region and were focused in perivenous zone 3 of all ethanol-fed animals. Protein and triglyceride levels were increased, whereas vitamin A and iron levels were decreased in liver homogenates from ethanol-fed animals. Thus, in this new animal model of alcoholism, ethanol feeding produced the features of alcoholic liver disease concurrent with hepatic deficiency of selected nutrients. Histological and immunofluorescent studies provide in vivo evidence that perivenous collagen deposition is linked to ethanol metabolism and acetaldehyde production. (HEPATOLOGY 1993;18:954-960).
What should we clinicians do for healthy, asymptomatic, hbsag-positive carriers?
Authors: McMahon Brian J.
Published: March 2007

Objective:

To assess the long-term outcome in hepatitis B surface antigen (HBsAg) carriers who have normal liver function tests, focusing on survival and the development of severe liver disease and hepatocellular carcinoma.

Design:

Cohort study with a mean follow-up of 130 months.

Setting:

Liver clinic of a referral center.

Patients:

Ninety-two HBsAg-positive blood donors with normal liver function tests.

Measurements:

Histologic evaluation of liver specimens at baseline; clinical, biochemical, and serologic follow-up; and repeat liver biopsy if clinically indicated or after 10 years of follow-up.

Results:

At baseline, 69 subjects had normal histologic findings or only minor abnormalities, 18 had chronic persistent hepatitis, and 5 had mild chronic active hepatitis. Serum enzyme levels remained normal in 58 of 68 patients who had regular follow-up. Three patients had biochemical changes consistent with hepatitis B virus (HBV) infection; in one of these patients, a later histologic evaluation showed progression to chronic active hepatitis. One patient developed alcoholic cirrhosis. Six other patients had mild or transient transaminase elevations, with no evidence of HBV replication, hepatitis D virus infection, hepatitis C virus (HCV) infection, or histologic deterioration. Liver histologic findings also remained unchanged in 21 patients who showed no biochemical changes during 10 years of follow-up and consented to have repeated liver biopsy. Ten patients showed loss of HBsAg; 2 of these patients acquired antibody to hepatitis B surface antigen (anti-HBs). All patients who did not have regular follow-up, except 1, were interviewed by telephone during 1990; All denied having liver disease. No patients developed hepatocellular carcinoma.

Conclusions:

Italian HBsAg carriers with initially normal liver function tests have an excellent prognosis: Delta superinfection is infrequent and the risk for developing hepatocellular carcinoma is low.
Southern-blot analysis and simultaneous In Situ detection of hepatitis B virus-associated DNA and antigens in patients with end-stage liver disease
Authors: Han Kwang Hyub , Hollinger Blaine F. , Noonan Christine A. , Solomon Harvey , Klintmalm Goran B. G. , Genta Robert M. , Yoffe Boris
Published: March 2007

To gain new insights into the pathogenesis of hepatitis B virus-induced chronic liver disease, we have used nonisotopic in situ detection methods for the simultaneous analysis of hepatitis B virus DNA and antigens at the single-cell level. Paraffin-embedded liver specimens from 23 cirrhotic patients (12 HBsAg positive and 11 HBsAg negative) who underwent liver transplantation were evaluated by in situ hybridization with a digoxigenin-labeled DNA probe and digoxigenin detection system and by immunohis-tochemistry with an enhanced biotin-streptavidin technique. DNAs extracted from liver and serum specimens were analyzed by Southern- and slot-blot hybridization, respectively. Using the in situ techniques, we detected hepatitis B virus-specific DNA and antigens in 11 of 12 HBsAg-positive patients and in none of the 11 HBsAg-negative individuals. Replicative intermediates of hepatitis B virus DNA were detected by Southern-blot analysis in the same 11 HBsAg-positive patients, 6 of whom had no serological markers of hepatitis B virus replication. Therefore a good correlation was found between the results obtained by the in situ and Southern-blot hybridization analyses of tissue specimens. However, a lack of correlation was found between serum- and tissue-associated markers of viral replication. In addition, the simultaneous in situ detection analyses revealed that some hepatocytes containing high levels of viral DNA were devoid of detectable HBcAg, suggesting a mechanism by which the virus may escape immunological surveillance. These data provide evidence that liver-associated HBV replication may persist in the absence of serological markers of active hepatitis B virus replication in cirrhotic patients with advanced liver disease and demonstrate that the evaluation of liver- rather than serum-associated markers of viral replication provides a more accurate assessment of the virological events occurring in HBsAg-positive individuals. (HEPATOLOGY 1993;18:1032-1038).
HLA B44-restricted cytotoxic T lymphocytes recognizing an epitope on hepatitis C virus nucleocapsid protein
Authors: Kita Hiroto , Moriyama Takashi , Kaneko Takashi , Harase Ichiro , Nomura Masayuki , Miura Hideaki , Nakamura Ikuo , Yazaki Yoshio , Imawari Michio
Published: March 2007

Cytotoxic T lymphocytes have been reported to be involved in the immune clearance of virus-infected cells and in the pathogenesis of viral infection. We studied the cytotoxic T lymphocyte response to the putative nucleocapsid protein of hepatitis C virus in patients with chronic hepatitis C. Cytotoxic T lymphocytes specific for hepatitis C virus nucleocapsid protein were generated from peripheral blood lymphocytes by means of repeated stimulation with a synthetic hepatitis C virus nucleocapsid protein peptide. The cytotoxic T lymphocytes were CD8 positive and recognized an epitope in hepatitis C virus nucleocapsid protein residues 81 to 100 in association with a human leukocyte antigen class I molecule, B44. The peptideinduced cytotoxic T lymphocytes recognized target cells synthesizing hepatitis C virus nucleocapsid protein endogenously, though less efficiently than peptide-pulsed target cells. The human leukocyte antigen B44-restricted cytotoxic T lymphocyte response was observed in three of five patients with chronic hepatitis C and a human leukocyte antigen B44 molecule but in neither of two hepatitis C virus-negative healthy individuals with human leukocyte antigen B44 molecules. The results demonstrate the presence of hepatitis C virus-specific cytotoxic T lymphocytes in the peripheral blood of patients with chronic hepatitis C and provide a strategy to study the role of cytotoxic T lymphocytes in the viral clearance and the pathogenesis of hepatitis C virus infection. (HEPATOLOGY 1993;18:1039-1044).
Prevention of posttransfusion hepatitis B and C by screening for antibody to hepatitis C virus and antibody to HBcAg
Authors: Chung Hau Tim , Lee Joseph Sai Kit , Lok Anna Suk Fong
Published: March 2007

Screening of blood donors by testing for antibody to HBcAg and antibody to hepatitis C virus is commonly done. However, the applicability of these screening tests may vary depending on the prevalence of hepatitis B virus and hepatitis C virus infection in various populations. We have therefore prospectively evaluated 158 adult patients who received blood or blood products during open-heart surgery in Hong Kong to compare the efficacy of various serological screening tests in the prevention of posttransfusion hepatitis. Serum from five (0.5%) donors was positive for antibody to hepatitis C virus by second-generation enzyme immunoassay; in two, optical-density readings in enzyme immunoassay were greater than 2.0, but only one was positive for hepatitis C virus RNA by reverse transcription-polymerase chain reaction. The latter donor was also positive for antibody to HBcAg and had elevated serum ALT activity. The recipient of a unit of this donor’s blood was the only one in whom posttransfusion hepatitis C developed (0.1% per unit transfused). Screening with antibody to hepatitis C virus was more specific than that with antibody to HBcAg or ALT in excluding donors from transmitting hepatitis C (99.6%, 79.4% and 98.8%, respectively). Both the sensitivity and negative predictive value of screening for antibody to hepatitis C virus were 100%, but the positive predictive value was only 20%.

Forty-five blood recipients were considered susceptible to hepatitis B virus infection because testing for hepatitis B serology in serum (HBsAg, antibody to HBsAg and antibody to HBcAg) was negative before being transfused. Asymptomatic hepatitis B seroconversion developed in three (6.7%) recipients (1.1% per unit transfused). One was transiently positive for HBsAg, and all three became positive for antibody to HBsAg and antibody to HBcAg during follow-up. One patient received a unit of blood that was positive for antibody to HBcAg and hepatitis B virus DNA by polymerase chain reaction. The second patient received a unit of blood with antibody to HBcAg but negative for hepatitis B virus DNA. The blood administered to the third patient was negative for hepatitis B markers, including hepatitis B virus DNA by polymerase chain reaction. Exclusion of donors with isolated antibody to HBcAg had the highest sensitivity and specificity for prevention of posttransfusion hepatitis B (66.7% and 96.1%, respectively). This approach yielded a positive predictive value of 16.7% and a negative predictive value of 99.6%. Antibody to hepatitis C virus should be screened in all blood donors to minimize the risk of posttransfusion hepatitis C. In hepatitis B virus-endemic areas, further reduction of the risk of posttransfusion hepatitis B with blood from HBsAg-negative volunteer donors is difficult. Our data suggest that excluding donors with isolated antibody to HBcAg may be the most effective approach. (HEPATOLOGY 1993;18:1045-1049).
Combined hepatocellular carcinoma and cholangiocarcinoma: Clinical features and computed tomographic findings
Authors: Aoki Kazunori , Takayasu Kenichi , Kawano Toshinobu , Muramatsu Yukio , Moriyama Noriyuki , Wakao Fumihiko , Yamamoto Junji , Shimada Kazuaki , Takayama Tadatoshi , Kosuge Tomoo , Yamasaki Susumu , Sakamoto Michiie , Hirohashi Setsuo
Published: March 2007

Clinicoradiological features were studied in 20 patients with 22 mass lesions of combined hepatocellular carcinoma and cholangiocarcinomas and findings of computed tomography in 12 of these patients with 14 hepatocellular carcinoma-cholangiocarcinomas. Five of these patients also had single overt hepatocellular carcinomas. The incidence of hepatocellular carcinoma-cholangiocarcinoma was 3.3% among the patients with primary liver cancer treated in our hospital. HBsAg was present in 25%, and increased levels of serum α-fetoprotein ( > 200 ng/ml) and carcinoembryonic antigen ( > 5 ng/ml) were found in 25% and in 47%, respectively. Associated cirrhosis was present in 60%. Analysis of 14 hepatocellular carcinoma-cholangiocarcinomas in 12 patients in whom the enhancement pattern on dynamic computed tomography and pathological findings could be studied and compared suggested three tumor types. Nine lesions (type A) were demonstrated only as areas with high-density peripheries in the early phase of enhancement that evolved into a pattern of peripheral low density and central high density in the late phase. Four masses (type B) were shown as hyperdense tumors (early phase) that changed to low density in the late phase. One mass (type C) was seen as a low-density lesion that did not change. Histopathologically, type A comprised hepatocellular carcinoma-predominant components in the peripheral area, cholangiocarcinoma-predominant components with abundant fibrous stroma in the central area and a tissue transitional between the two in the midzone. By contrast, two of four type B masses comprised hepatocellular carcinoma with scattered cholangiocarcinoma components throughout the tumor. The remaining two masses had main nodules of hepatocellular carcinoma and adherent small cholangiocarcinomas. One type C mass had a main cholangiocarcinoma nodule and an adjacent small hepatocellular carcinoma. Differentiation of hepatocellular carcinoma-cholangiocarcinoma from hepatocellular carcinoma or cholangiocarcinoma with computed tomography alone was difficult. When computed tomography results and α-fetoprotein or carcinoembryonic antigen levels show some discrepancy, the possibility of hepatocellular carcinoma-cholangiocarcinoma should be considered. (HEPATOLOGY 1993;18:1090-1095).
Resolution of liver biopsy alterations in three siblings with bile acid treatment of an inborn error of bile acid metabolism (Δ4-3-oxosteroid 5β-reductase deficiency)
Authors: Daugherty Ynthia C. , Setchell Kenneth D. R. , Heubi James E. , Ballstreri William F.
Published: March 2007

Identical male twins and their brother, cholestatic from birth, with Δ4-3-oxosteroid 5δ-reductase deficiency, were studied by serial liver biopsy. Spectrometry documented defective primary bile acid synthesis and markedly increased levels of atypical oxo and allo bile acids in urine and serum. Hepatocellular cholestásis and giant-cell transformation resolved in parallel with clinical and biochemical recovery during oral bile acid administration. In the twins, portal fibrosis stabilized at a mild level; they are well as 5-yr-olds at this writing. Follow-up biopsy in their brother at 8 mo was normal, and he is doing well at 3 yr of age. Hepatic ultrastructural alterations in all three were characterized by abnormalities of bile canaliculi including small bile plugs, diverticulae and latticelike elaborations of hepatocellular membranes adjacent to bile canaliculi that were shown to have resolved completely on subsequent biopsies. Eight additional cases have been detected on urine screening; only two of these patients have survived, on bile acid therapy. Early diagnosis and treatment improves the prognosis of this otherwise lethal inborn error of bile acid synthesis. (HEPATOLOGY 1993; 18:1096-1101).
Randomized controlled trial of desmopressin plus terlipressin vs. terlipressin alone for the treatment of acute variceal hemorrhage in cirrhotic patients: A multicenter, double-blind study
Authors: de Franchis Roberto , Arcidiacono Paolo G. , Carpinelli Luca , Andreoni Bruno , Cestari Lorenzo , Brunati Sergio , Zambelli Alessandro , Battaglia Giorgio , Mannucci Pier M.
Published: March 2007

1-Deamino-8-D-arginine vasopressin (DDAVP, desmopressin), a synthetic analog of the antidiuretic hormone l-arginine vasopressin, improves hemostasis parameters in cirrhotic patients. Hence its use in combination with a vasoactive drug such as terlipressin might improve the performance of this drug in controlling variceal bleeding. The aim of this trial was to compare the efficacy of desmopressin plus terlipressin with that of terlipressin alone in controlling acute variceal hemorrhage. Cirrhotic patients with active variceal hemorrhage diagnosed endoscopically were randomized within 2 hr of admission to receive desmopressin plus terlipressin or placebo plus terlipressin. Terlipressin (2 mg, intravenous bolus) was given at time 0 and every 4 hr thereafter for 24 hr. Desmopressin (0.3 μg/kg, intravenously) or placebo was given in saline solution over 30 min at time 0 and at 26 hr. Patients were monitored for 24 hr after cessation of treatment. Treatment failure was defined as recurrence of active bleeding during treatment or within the 24 hr after treatment. After enrolling 51 of the planned 84 patients, we carried out an interim analysis. Treatment failure occurred in 13 of 24 patients randomized to receive desmopressin plus terlipressin (54.2%) and in 6 of 22 patients randomized to receive terlipressin (27.3%) (p = 0.06, Fisher’s exact test). The trial was interrupted at this stage because patients treated with the “new” therapy fared worse than those treated with the standard therapy, and the possibility of reversing this trend by completing the trial was deemed remote. The addition of desmopressin does not improve and may worsen the efficacy of terlipressin in controlling acute variceal bleeding in cirrhotic patients. (HEPATOLOGY 1993;18:1102-1107).
Steroid receptors and heat-shock proteins in patients with primary biliary cirrhosis
Authors: Jorge Arturo D. , Stati Arturo O. , Roig Laura Vargas , Ponce Guillermo , Jorge Oliver A. , Ciocca Daniel R.
Published: March 2007

Primary biliary cirrhosis has a definite female preponderance. Increased estrogen levels have been found in patients with this disease; however no studies indicate the status of sex hormone steroid receptors in primary biliary cirrhosis patients. In this study the occurrence and distribution of estrogen receptors, progesterone receptors and androgen receptors in liver biopsy specimens from patients with primary biliary cirrhosis were examined and compared with these receptors in the normal liver. In addition, three heat-shock proteins associated with steroid receptors (90 kD, 70 kD and 27 kD) were examined. All of the receptor proteins were detected on immunocytochemical study using specific receptor antibodies; monoclonal and polyclonal antibodies were also used to detect the heat-shock proteins. Normal bile duct epithelial cells displayed low-to-moderate amount of estrogen receptors and abundant 90-kD, 70-kD and 27-kd heat-shock protein expression, whereas normal hepatocytes showed moderate estrogen receptor and 90-kD heat-shock protein and high 70-kD heat-shock protein expression. Expression of 70-kD heat-shock protein was due mainly to the constitutive form of this protein (hsc72). In patients with primary biliary cirrhosis, significant increases in estrogen receptor and 90-kD heat-shock protein content were seen in bile duct cells and in hepatocytes. Levels of 27-kD heat-shock protein were also increased in some of the primary biliary cirrhosis biopsy specimens. The expression of progesterone receptor and androgen receptor was very low in normal and primary biliary cirrhosis bile duct cells and hepatocytes. The high expression of estrogen receptor and associated heat-shock proteins in primary biliary cirrhosis patients indicates that estrogens and their receptors may play a role in this disease, a finding that may have therapeutic implications. (HEPATOLOGY 1993;18:1108-1114).
Increased serum nitrite and nitrate levels in patients with cirrhosis: Relationship to endotoxemia
Authors: Guarner Carlos , Soriano German , Tomas Albert , Bulbena Oriol , Novella Maria Teresa , Balanzo Joaquin , Vilardell Francisco , Mourelle Marisabel , Moncada Salvador
Published: March 2007

Nitric oxide derived from vascular endothelium is a potent vasodilator that plays a key role in the homeostasis of blood pressure. Because cirrhotic patients tend to have low arterial pressure, we measured in 51 patients and 10 control subjects serum nitrite and nitrate levels as an index of in vivo nitric oxide generation. We also measured plasma endotoxin, a substance frequently increased in cirrhotic patients and known to induce nitric oxide synthesis. Cirrhotic patients showed significant increases in serum nitrite/nitrate and plasma endotoxin compared with controls. Values were particularly increased in patients with decompensated cirrhosis, as manifested by ascites with or without functional kidney failure. High serum nitrite/nitrate levels were associated with high plasma renin activity, high aldosterone and antidiuretic hormone levels and low urinary excretion of sodium. In addition, serum nitrite/nitrate levels significantly correlated with endotoxemia. Oral administration of colistin to 15 cirrhotic patients reduced significantly plasma endotoxin levels (p < 0.01) and serum nitrite/nitrate levels (p < 0.05). Because endotoxin enhances the expression of inducible nitric oxide synthase, our results suggest that circulating endotoxin in cirrhosis is responsible for excessive synthesis and release of nitric oxide by the vasculature. These findings might explain the hemodynamic dysfunction seen in cirrhotic patients. (HEPATOLOGY 1993;18:1139-1143).
Experimental evidence that the physiological position of the liver within the circulation is not a major determinant of zonation of gene expression
Authors: Wagenaar Gerry T. M. , Chamuleau Robert A. F. M. , de Haan Jan G. , Maas Martinus A. W. , de Boer Piet A. J. , Marx Frans , Moorman Antoon F. M. , Frederiks Wilma M. , Lamers Wouter H.
Published: March 2007

Many enzymes are distributed heterogeneously within the liver lobule. The factors that play a determining role in the establishment and maintenance of these heterogeneous expression patterns have not yet been identified. To investigate whether the composition of the afferent hepatic blood plays a crucial role in the maintenance of the heterogeneity of gene expression of the parenchymal cells within the liver lobule, we changed the source of the afferent hepatic blood by microsurgical techniques. Three different groups of experimental animals were studied: rats with livers that are perfused with portal blood only (ligation of the hepatic artery), with caval blood only (portocaval transposition and ligation of the hepatic artery) and arterial blood only (portocaval shunt, arterialization of the distal end of the portal vein and ligation of the hepatic artery). To study differences in gene expression patterns, we chose enzymes that have a heterogeneous expression pattern within the liver lobule: the periportally located enzymes carbamoylphosphate synthase, succinate dehydrogenase, phosphoenolpyruvate carboxylkinase and the pericentrally located enzymes glutamine synthase, glutamate dehydrogenase and NADPH-cytochrome P-450 reductase. To eliminate the potential interference of the long half-lives of some of these proteins on the interpretation of the results, we also studied the distribution of the mRNAs of carbamoylphosphate synthase, glutamine synthase, glutamate dehydrogenase and phosphoenolpyruvate carboxykinase. The animals were studied 2 wk after the operations. On the basis of their changes in body weight the animals were in steady state for at least a week. The patterns of gene expression of the enzymes studied did not change, regardless of the source of the altered afferent hepatic blood. The changes in gene expression that were observed in animals that did not regain their preoperative weight were shown to be caused by a limited intake of food. This study demonstrates that the physiological position of the liver within the circulation (i.e., between the gastrointestinal tract and the systemic circulation) is not as critical as is often stated and is certainly not essential for the maintenance of liver cell heterogeneity. The data suggest that the direction of the bloodstream (i.e., the existence of an upstream and a downstream compartment) is a major determinant of zonation of gene expression. (HEPATOLOGY 1993;18:1144-1153).
Parallel decrease of Na+ -taurocholate cotransport and its encoding mRNA in primary cultures of rat hepatocytes
Authors: Liang Dana , Hagenbuch Bruno , Stieger Bruno , Meier Peter J.
Published: March 2007

We investigated the molecular mechanism underlying the progressive loss of Na+ -dependent bile salt uptake in primary cultured rat hepatocytes. A specific cDNA probe was used to quantitate the levels of mRNA encoding the Na+ -taurocholate-cotransporting polypeptide at various culture times. Hepatocytes were cultured on collagen in the presence of insulin (10−7 mol/L), dexamethasone (10−7 mol/L) and 10% fetal calf serum for up to 72 hr. During this time period the dissociation constant of Na+ -dependent taurocholate uptake remained stable (19 to 39 μmol/L), whereas the maximum velocity values decreased from 100% at 3 hr to 55%, 22% and 4% at 24, 48 and 72 hr, respectively. Concomitantly the levels of the Na+ -taurocholate-cotransporting polypeptide mRNA also decreased from 100% at 3 hr to 41%, 24% and 4% at the later time points. In contrast, Northern hybridization with complementary DNA probes for three common housekeeping gene products revealed a 1.8- to 3.4-fold increase in the levels of mRNA encoding the α-subunit of the Na+K+ -ATPase, β-actin and glycerol-3-phosphate dehydrogenase. These data indicate that the loss of Na+ -dependent bile salt uptake in primary cultures of rat hepatocytes is caused by decreased levels of its specific mRNA. Hence the studies further confirm that without specific measures (primary) cultured rat hepatocytes reverse their liver-specific phenotype to a more fetal pattern of gene expression. (HEPATOLOGY 1993;18:1162-1166).
Analysis of glucocorticoid receptors in human hepatocellular carcinoma and HepG2 cells
Authors: Lui Wing-Yiu , P’Eng Fang-Ku , Chang Yuh-Fang , Chang Tai-Jay , Tsai Ting-Fan , Hsu Mei-Ling , Su Tsung-Sheng , Tsay Shyh-Haw , Wu Chew-Wun , Liu Tsung-Yun , Chi Chin-Wen
Published: March 2007

Hepatocellular carcinoma is the leading cause of male cancer death in Taiwan. We have found that the level of glucocorticoid receptor in hepatocellular carcinoma is significantly higher than that in the peritumoral tissue. In this study, we used a rat liver glucocorticoid receptor complementary DNA probe to examine the expression of glucocorticoid receptor gene in 15 paired samples of hepatocellular carcinoma and their peritumoral tissues. No differences in genomic DNA patterns of the glucocorticoid receptor gene were found between the tumor and peritumoral tissues. The amount of glucocorticoid receptor was found to be significantly higher in hepatoma samples than in peritumoral liver samples. The levels of glucocorticoid receptor messenger RNAs were increased in most tumors compared with their peritumoral samples. To examine the function of glucocorticoid receptors in hepatoma, we examined the expression of glucocorticoid receptor and its relation to cell-cycle progression in human HepG2 cells. Using specific monoclonal antibodies and flow cytometric study, we found glucocorticoid receptor to be expressed constitutively in all cell-cycle phases. In addition, hydrocortisone treatment of HepG2 cells resulted in increased expression of glucocorticoid receptors and increased secretion of α-fetoprotein. RU-486, a glucocorticoid antagonist, blocked the hydrocortisone effect, indicating that glucocorticoid receptors are functional in HepG2 cells. Taken together, our data suggest that glucocorticoids and their receptors play an important role in the growth of hepatoma. (HEPATOLOGY 1993;18:1167-1174).
Bile acid-induced modifications in DNA synthesis by the regenerating perfused rat liver
Authors: Marin Jose J. G. , Barbero Emilio R. , Herrera Maria C. , Tabernero Arantxa , Monte Maria J.
Published: March 2007

Liver cell proliferation is a complex process that can be affected by a large number of factors such as bile acids, which have been reported to be associated to the pathogenesis of liver cancer. In this work, bile acid-induced modifications in DNA synthesis by regenerating perfused rat liver were investigated. Two-thirds hepatectomy was carried out 24 hr before perfusion of livers with recirculating, erythrocyte-free Krebs-Henseleit solution. The viability of the preparations was maintained under all experimental conditions, as indicated by bile flow, oxygen uptake, perfusion pressure, perfusion flow and release of lactate dehydrogenase and potassium into the perfusate. Livers received (min 10 to min 60) bile acid infusion at a rate of 25 nmol/min/gm liver (i.e., maximal secretion rate/2) in regenerating livers as calculated for taurocholate in separate experiments). Trace amounts of [methyl-14C]thymidine were added to the perfusate at min 30. At the end of the experiments (min 60) the livers were washed, removed, weighed and homogenized to determine radioactivity in whole tissue, in DNA and in non-DNA-related fractions. Taurocholate and, to a lesser extent, taurodeoxycholate and dehydrocholate (but not ursodeoxycholate) were found to reduce 14C incorporation into DNA. This was not due to changes in the content of 14C in whole, regenerating liver tissue. Taurocholate, taurodeoxycholate, dehydrocholate and ursodeoxycholate had no effect on thymidine uptake; moreover, the proportion of 14C found in bile was negligible. However, bile acid-induced modification in the fate of intracellular thymidine was observed. In regenerating livers receiving no bile acid, the 14C carried by thymidine metabolites accounted for about 60% of 14C in whole liver tissue. Taurocholate markedly increased this proportion to about 80%. Reverse-phase high-pressure liquid chromatography revealed that most of this 14C (about 80%) was recovered at the elution time, corresponding to thymidine catabolites rather than to DNA precursors. These results suggest that bile acids induce enhancement of thymidine catabolism that reduces its incorporation into DNA; inhibition in the process of DNA synthesis itself, leading to a subsequent increase in the metabolism of DNA precursors; or both. Moreover, from the diversity in this property for bile acid species it might be inferred that changes in the composition and size of the bile acid pool during liver carcinogenesis or regeneration play a role in the modulation of the proliferative process. (HEPATOLOGY 1993;18:1182-1192).
Induction of metallothionein and its localization in the nucleus of rat hepatocytes after partial hepatectomy
Authors: Tohyama Chiharu , Suzuki Junko Sayato , Hemelraad Jack , Nishimura Noriko , Nishimura Hisao
Published: March 2007

Metallothioneins, a group of cysteine-rich heavymetal binding proteins, are induced in the regenerating rat liver in response to the stimuli evoked by partial hepatectomy. We have investigated the expression of metallothionein genes and proto-oncogenes (c-fos, c-jun and c-myc), as well as specific localization of metallothionein in the liver cells after partial hepatectomy. Metallothionein mRNA was detected as early as 3 hr and reached a maximal level by 6 hr. Expression of the proto-oncogenes apparently preceded the elevation of metallothionein protein because the latter was maximal 18 hr after partial hepatectomy, followed by a decrease until 70 hr. Hepatocytes of the intact rat liver have metallothionein in the cytoplasm only. Interestingly, metallothionein was localized predominantly in the nucleus as early as 6 hr after partial hepatectomy, and the staining intensity of metallothionein became maximal at 15 hr, followed by detection in both the cytoplasm and nucleus at 24 hr or longer. The use of a confocal laser scanning microscope with both tissue sections and isolated nuclei has clearly shown that metallothionein immunofluorescence exists inside hepatocyte nuclei after partial hepatectomy. Expression of the proto-oncogenes c-fos and c-jun is elevated after partial hepatectomy, and the resultant heterodimer of gene products may contribute to the observed metallothionein gene induction. However, the observation that metallothionein protein levels were elevated until 18 hr after partial hepatectomy suggests that an alternative pathway for the induction of metallothionein gene expression may also be present. It is proposed that the specific localization of metallothionein in the nucleus may be involved in the early events associated with the compensatory cell proliferation that follows partial hepatectomy. (HEPATOLOGY 1993;18:1193-1201).
Inhibitory effect of gastrin and cholecystokinin fragments on the secretion of triacylglycerol in rat hepatocytes
Authors: Lindholm Marie , Eklund Anders
Published: March 2007

In this investigation we studied the influence of two gastrin fragments, pentagastrin and nonsulfated heptadecagastrin, and two cholecystokinin fragments, sulfated and desulfated cholecystokinin 26-33, on intracellular and secreted triacylglycerol in isolated hepatocyte cultures. Both gastrin fragments inhibited triacylglycerol release in a biphasic manner, exhibiting maximal effect at 0.1 nmol/L (nonsulfated heptadecagastrin) and 0.3 nmol/L (pentagastrin). At these concentrations triacylglycerol secretion was 42% (non-sulfated heptadecagastrin, p < 0.001) and 62% (pentagastrin, p < 0.001) lower than in cells untreated with gastrin. Sulfated cholecystokinin 26-33 caused a 35% decrease in triacylglycerol secretion at 0.1 nmol/L (p < 0.01), and desulfated cholecystokinin 26-33 caused a 53% decrease at 0.2 nmol/L (p < 0.001). In all experiments, the hormone-induced decrease in triacylglycerol secretion was accompanied by an increase in intracellular triacylglycerol content. The cholecystokinin-A receptor antagonist L-364, 718 did not affect the decrease in triacylglycerol secretion induced by 0.3 nmol/L pentagastrin, whereas the cholecystokinin-B receptor antagonist L-365, 260 inhibited the pentagastrin effect at concentrations above 50 nmol/L. These results suggest that gastrin, cholecystokinin or some other gastrinlike hormone (or all three) may play a previously unrecognized regulatory role with respect to hepatic very low density lipoprotein secretion. (HEPATOLOGY 1993;18:1232-1237).