bluuebunny/crossref_metadata_2025 huggingface dataset:
Reduction of morbidity and mortality from biliary complications after liver transplantation
Authors: Klein Andrew S. , Savader Scott , Burdick James F. , Fair Jeffrey , Mitchell Mack , Colombani Paul , Perler Bruce , Osterman Floyd , Williams Melville G.
Published: March 2007

Over a 4-yr period that began October 1, 1986, 103 orthotopic liver transplants were performed on 91 patients at the Johns Hopkins Hospital. Biliary reconstruction at the time of transplantation was performed in standard fashion by an appropriately trained member of the surgical team. Six (7%) patients developed biliary complications, which included three cases of common bile duct stricture and one case each of bile duct obstruction caused by biopsy-related hemobilia, biloma and a retained fragment of a T tube after removal. Five of the six patients were treated successfully by nonoperative interventional radiological procedures performed under local anesthesia with light intravenous sedation. Reoperative surgery (to remove the T-tube fragment from subcutaneous tissue) was required for only one patient, and no deaths (0 of 91) were attributable to biliary complications. All six patients are alive and well 6 to 33 mo after the operation with excellent liver function. Our findings would suggest that most biliary complications of orthotopic liver transplants are avoidable and that the few that do appear can usually be managed both safely and effectively by an interventional radiological approach. (Hepatology 1991;14:818-823).
Effect of propranolol on portosystemic collateral circulation in patients with cirrhosis
Authors: Gaiani Stefano , Bolondi Luigi , Fenyves Daphna , Zironi Gianni , Rigamonti Alessandra , Barbara Luigi
Published: March 2007

Propranolol has been demonstrated to be effective in lowering portal pressure in cirrhotic patients. This effect is mediated by a reduction of splanchnic arterial inflow and a consequent decrease of portal vein and portocollateral blood flow. Although experimental studies suggest a direct effect of the drug on portocollateral circulation, little information exists about relative flow changes occurring in the portal vein and in collateral veins feeding esophageal varices. This study addressed the problem in 12 cirrhotic patients selected on the basis of feasibility of Doppler flowmetry in both the portal and left gastric veins. Caliber, flow velocity and flow volume in both vessels were measured by Doppler ultrasound before and at 60, 120 and 180 min after an oral dose of 40 mg propranolol, together with heart rate and mean arterial pressure.

A significant decrease in heart rate (− 17.6% ± 1.1%, p < 0.001) and mean arterial pressure (− 10.6% ± 0.9%, p < 0.005) confirmed effective β-blockade. Baseline flow velocity was significantly lower in the portal vein than in the left gastric vein (12.4 ± 0.6 vs. 15.4 ± 1.5 cm/sec, p < 0.05). Maximal hemodynamic effect was reached at 120 min after administration of propranolol. The vessel caliber did not change significantly. Flow velocity fell from 12.4 ± 0.6 to 10.4 ± 0.7 cm/sec in the portal vein (p < 0.05) and from 15.4 ± 1.5 to 11.1 ± 0.9 cm/sec in the left gastric vein (p < 0.01). The percentage decrease in velocity and flow volume was significantly higher in the left gastric vein than in the portal vein (velocity = − 26.8% vs. − 15%, p < 0.05; flow = − 30.2 ± 4.4 vs. − 20.9 ± 3.3 cm/sec; p < 0.05). These data support the hypothesis of a direct effect of the drug on portocollateral resistance. (Hepatology 1991;14:824-829).
Ursodeoxycholic acid for the treatment of primary sclerosing cholangitis: A 30-month pilot study
Authors: O’brien Christopher B. , Senior John R. , Arora-Mirchandani Renu , Batta Ashok K. , Salen Gerald
Published: March 2007

We investigated the effects of once-daily oral administration of 10 mg/kg ursodeoxycholic acid (generic name, ursodiol) on elevated serum enzyme activities, bilirubin, cholesterol, bile acids and symptoms in patients with primary sclerosing cholangitis. A 30-mo, open-label, pilot trial was designed to cover four periods: (a) 3 mo of pretreatment observation (period 1), (b) 6 mo on ursodiol (period 2), (c) 3 mo withdrawal of treatment (period 3) and (d) 18 mo of extended retreatment (period 4). Diagnosis was confirmed by cholangiography and liver biopsy specimens. We enrolled 12 patients with persistently elevated pretreatment alkaline phosphatase and γ-glutamyltransferase levels (at least twice the upper limit of normal), and observed them for a median of 37 mo.

Significant reductions in serum total cholesterol levels and in serum enzyme activities indicating cholestasis and hepatocellular injury occurred during ursodiol treatment in both treatment periods 2 and 4 and relapsed with treatment interruption in period 3. Elevated serum bilirubin and symptoms of disabling fatigue, pruritus and diarrhea were improved by ursodiol. Improvements have continued after 2 yr of treatment in 10 patients (1 patient had a transplantation after he relapsed on withdrawal of ursodiol therapy; another died of postoperative complications of colon resection for carcinoma). No other cases of clinical deterioration were observed in the retreatment period. The longer term reductions of alkaline phosphatase, transaminases, bilirubin and cholesterol after 2 yr of treatment were even greater than the initial reductions after 6 mo of treatment. These results justify initiation of larger, controlled clinical trials, with serial morphological evaluations of the liver and biliary tree. (Hepatology 1991;14:838-847).
Localization of CYP1A1 and CYP1A2 messenger RNA in normal human liver and in hepatocellular carcinoma by in situ hybridization
Authors: McKinnon Ross A. , Hall Pauline de la M. , Quattrochi Linda C. , Tukey Robert H. , McManus Michael E.
Published: March 2007

To better characterize the precise cellular distribution of CYP1A gene products in man, we have undertaken Northern-blot and in situ hybridization analyses of CYP1A expression in human liver. Using riboprobes transcribed from both CYP1A1 and CYP1A2 complementary DNAs to probe a series of Northern blots of 23 human liver messenger RNA samples, CYP1A1 expression was demonstrated in 11 samples and CYP1A2 expression was evident in 22 samples. The level of expression of both CYP1A enzymes in these livers demonstrated marked variability. The CYP1A1 and CYP1A2 riboprobes were then used for in situ hybridization localization of CYP1A1/1A2 messenger RNA sequences on paraffin-embedded, formalin-fixed human liver sections. These studies demonstrated that both CYP1A1 and CYP1A2 messenger RNAs are distributed nonuniformly across the human liver acinus, with levels highest in hepatocytes surrounding terminal hepatic venules and intercalated veins. Immunohistochemistry with an anti-rabbit CYP1A1 serum demonstrated a corresponding distribution for the translated CYP1A proteins. In situ hybridization analysis was also performed on sections of hepatocellular carcinoma, demonstrating a significant down-regulation in CYP1A expression. Functional studies using the activation of the food-derived heterocyclic amine MeIQ (2-amino-3,4-dimethylimadazo [4,5-f] quinoline) to a mutagen in the Ames test as an indicator of CYP1A expression confirmed this down-regulation. These results demonstrate heterogeneity of hepatic CYP1A expression both between individuals and in different acinar zones. This variation in expression may be of significance in assessing cell specific toxicities of various drugs and carcinogens. (Hepatology 1991;14:848-856).
Dexamethasone modulates α2-macroglobulin and apolipoprotein E gene expression in cultured rat liver fat-storing (Ito) cells
Authors: Ramadori Giuliano , Knittel Thomas , Schwögler Stefan , Bieber Florian , Rieder Hartmut , Büschenfelde Karl-Herrmann Meyer Zum
Published: March 2007

Fat-storing (Ito) cells are perisinusoidal liver cells thought to play a central role in vitamin A metabolism and fibrogenesis. Glucocorticoids have been shown to be beneficial in the treatment of certain types of liver diseases by delaying the development of cirrhosis. To study the regulatory effects of dexamethasone on Ito cell gene expression, Ito cells were isolated from normal rat liver and primary cultures were established. The effect of dexamethasone on the synthesis of α2-macroglobulin, apolipoprotein E, fibronectin and actin was examined. Protein synthesis was studied both at the protein level and at the RNA level by means of biosynthetic labeling, immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by Northern blot analysis of total RNA.

After exposure to dexamethasone for 20 hr, α2-macroglobulin protein synthesis was increased threefold, whereas apolipoprotein E expression was decreased 80%. Biosynthesis of fibronectin remained unaffected by hormone treatment. The dexamethasone effect became detectable 5 hr after beginning the exposure. Deinduction kinetic experiments showed that the glucocorticoid effect was detectable more than 12 hr after the replacement of the dexamethasonecontaining culture medium by medium without the hormone.

Corresponding to the data obtained at the protein level, dexamethasone increased the steady-state levels of α2-macroglobulin-specific messenger RNA and reduced apolipoprotein E-specific transcripts, whereas fibronectin and actin messenger mRNA remained unchanged. These findings indicate that glucocorticoids can affect the Ito-cell gene expression in a differentiated manner. Because dexamethasone not only stimulates α2-macroglobulin but also fibronectin and apolipoprotein E gene expression in rat hepatocytes, it will be interesting to study the difference in regulation of those genes in both cell populations. (Hepatology 1991;14:875-882).
Kupffer cells from carbon tetrachloride-injured rat livers produce chemotactic factors for fibroblasts and monocytes: The role of tumor necrosis factor-α
Authors: Armendariz-Borunda Juan , Seyer Jerome M. , Postlethwaite Arnold E. , Kang Andrew H.
Published: March 2007

Conditioned media from cultured Kupffer and mononuclear macrophagic cells obtained 48 hr after CCl4 administration to rats contains chemotactic factors for human skin fibroblasts and human monocytes. The chemotactic mediator for fibroblasts was approximately 17 kD and was more prominent at early stages of culture. It induced a dose-dependent chemotactic response in fibroblasts. Although the conditioned medium from cultured Kupffer cells of normal rats also contained detectable biological activity, it was significantly less than that in conditioned medium from cultured Kupffer cells from CCl4-treated rats. The activity obtained after purification by highperformance liquid chromatography was completely ablated by incubation with tumor necrosis factor-α antibody. Transforming growth factor-β antibody diminished biological activity by 20%. Human recombinant tumor necrosis factor-α and transforming growth factor-β used in the assay as control showed significant chemotactic activity. The chemotactic activity present in whole normal conditioned medium was only present after 24 and 48 hr of culture. Furthermore, this activity was not neutralized by human recombinant tumor necrosis factor-α or transforming growth factor-β antibodies. Incubation of whole 6-hr conditioned medium with human recombinant tumor necrosis factor-α and transforming growth factor-β antibodies demonstrated and confirmed that tumor necrosis factor-α plays a major role in inducing the chemotactic response. On acidification of this supernatant, we found a notable increase in the biological response that could be neutralized by transforming growth factor-β antibody. Thus tumor necrosis factor-α and transforming growth factor-β may sequentially provide important signals for fibroblast and monocyte recruitment in vivo at initial stages of liver injury. (Hepatology 1991;14:895-900).
Fuel homeostasis and carnitine metabolism in rats with secondary biliary cirrhosis
Authors: Krahenbuhl Stephan , Brass Eric P.
Published: March 2007

Energy metabolism is abnormal in patients and experimental animals with liver cirrhosis. To help better understand the abnormalities, fuel homeostasis and carnitine metabolism were studied in fed and 24-hr-starved rats with secondary biliary cirrhosis induced by bile duct ligation for 4 wk. Plasma ketone body concentrations were decreased by 67% in starved, bile duct-ligated rats compared with control rats. In contrast, plasma nonesterified fatty acid concentrations were not different between bile duct-ligated and control rats in the fed or the fasted state. Plasma triglyceride concentrations showed the expected decrease with starvation in control rats, but were increased with starvation in bile duct-ligated rats. Urinary excretion of dicarboxylic acids was increased in both fed and fasted bile duct ligated-rats compared with the respective control groups. Compared with control rats, hepatic total carnitine content (per gram of liver) was increased by 24% in fed and by 36% in fasted, bile duct-ligated rats. Fed, bile duct-ligated rats had an increased short-chain acylcarnitine-tocarnitine ratio in liver, plasma and urine compared with control rats. Analysis of the hepatic coenzyme A pool showed decreased coenzyme A content in fed and fasted bile duct-ligated rats compared with control rats. Hepatic long-chain acylcarnitine and long-chain acyl-coenzyme A content increased with starvation both in control and bile duct-ligated rats. The rise in plasma nonesterified fatty acid concentration and hepatic long-chain acylcarnitine and long-chain acylcoenzyme A contents with starvation in bile duct-ligated rats are consistent with unaltered hepatic availability of fatty acids. Thus bile duct-ligated rats demonstrated blunted ketosis during starvation and altered hepatic lipid metabolism. Bile duct-ligated rats represent an animal model for the study of fuel homeostasis and carnitine metabolism in the presence of chronic cholestatic liver disease. (Hepatology 1991;14:927-934).
The influence of propanolol on portosystemic shunting
Authors: Gumucio Jorge J. , Blendis Laurence M.
Published: March 2007

We investigated the role of early portal hypotensive pharmacotherapy in preventing the development of portal-systemic shunting in a portal hypertensive model of chronic murine schistosomiasis induced by infecting C3H mice with 60 cercariae of Schistosoma mansoni. Propranolol was administered in drinking water to 20 animals for a period of 6 wk at a dose of 10 mg · kg−1 d−1, starting at 5 wk of schistosomal infection. 32 age-matched mice with chronic schistosomal infection served as controls. All animals were studied 11 wk after the infection. Compared with controls the portal pressure (10.8 ± 0.40 mm Hg) was significantly lower (P < 0.001) in the propranolol-treated animals (7.9 ± 0.80 mm Hg). Portal-systemic shunting was decreased by 79%, from 12.2 ± 3.34% in controls to 2.5 ± 0.99% in the propranolol group (P < 0.05). Portal venous inflow was reduced by 38% in the propranolol treated animals (2.50 ± 0.73 ml/min; n = 6) compared with controls (4.00 ± 0.34 ml/min; n = 8; P < 0.05). The worm burden, the granulomatous reaction, the collagen content of the liver, and the serum bile acid levels were not significantly different between the two groups of animals. These results demonstrate that in chronic liver disease induced by schistosomiasis, the development of portal-systemic shunting can be decreased or prevented by the reduction of flow and pressure in the portal system.
Effect of interferon-γ on hepatitis B viral antigen expression in primary hepatocyte culture†
Authors: Lau Johnson Y. N. , Bain Vincent G. , Naoumov Nikolai V. , Smith Heather M. , Alexander Graeme J. M. , Williams Roger
Published: March 2007

Interferon-α has been shown recently to selectively enhance hepatocyte expression of HBsAg/pre-S2 in chronic hepatitis B virus infection in a way that may enhance immune recognition. To determine the effect of interferon-γ on hepatitis B virus antigen expression, hepatocytes isolated from patients with chronic hepatitis B virus infection were incubated in the absence or presence of interferon-γ and viral antigen expression was assessed by both radioimmunoassay and immunocytochemistry using appropriate monoclonal antibodies.

Interferon-γ inhibited the expression of all hepatitis B virus antigens tested. Intracellular HBsAg measured by radioimmunoassay of sonicated hepatocytes fell by 29% with 1 U/ml (p < 0.01) and 36% with 10 U/ml of interferon-γ (p < 0.001) compared with control treatment. Secreted HBsAg was reduced by 19% with 10 U/ml of interferon-γ (p < 0.01). Intracellular HBeAg was also decreased by 29% with 1 U/ml (p < 0.05) and 42% with 10 U/ml of interferon-γ (p < 0.05), but no significant change was found in the amount of secreted HBeAg. The proportion of hepatocytes containing various hepatitis B virus antigens and the intracellular viral antigen staining densities also fell significantly with interferon-γ incubation. Interestingly, the addition of interferon-γ abolished the augmenting effect of interferon-α on intracellular HBsAg.

These data indicate that interferon-γ, in contrast to interferon-α, has an inhibitory effect on hepatocyte expression of all hepatitis B virus antigens including HBsAg/pre-S2, suggesting that this may be one factor that accounts for their difference in clinical activity in patients with chronic hepatitis B virus infection. (Hepatology 1991;14:975-979.)
Autoantibody against dihydrolipoamide dehydrogenase, the E3 subunit of the 2-oxoacid dehydrogenase complexes: Significance for primary biliary cirrhosis
Authors: Maeda Takashi , Loveland Bruce E. , Rowley Merrill J. , Mackay Ian R.
Published: March 2007

Autoantibodies in primary biliary cirrhosis recognize mitochondrial 2-oxacid dehydrogenase complexes, particularly the E2 subunits. Reactivity with the E3 subunit, common to each of the enzyme complexes, was sought by immunoblotting, with sera screened at 1:100 instead of the conventional 1:1,000 dilution. This was found in 11 of 29 sera from patients with primary biliary cirrhosis but also in 10 of 40 sera from normal subjects. Two-dimensional immunoblotting and immunoblotting on purified enzymes established that the reactivity was actually with E3 rather than with another component of the 2-oxoacid enzymes of similar molecular weight. Purified antibodies to E3 eluted from an affinity column did not cross-react with other components of the 2-oxoacid enzyme complexes. The antibodies to E3 did not react with the Escherichia coli or yeast E3 subunits, suggesting that they are not stimulated by immune responses against microorganisms. Thus the proposal that reactivity to the shared E3 subunit of the 2-oxoacid enzyme complexes could initiate primary biliary cirrhosis is not reflected at the antibody level. (Hepatology 1991;14:994-999.)
Plasma metallothionein concentration in patients with liver disorders: Special emphasis on the relation with primary biliary cirrhosis
Authors: Mulder Theo P. J. , Janssens Roelof A. , Verspaget Hein W. , Lamers Cornelis B. H. W.
Published: March 2007

The plasma metallothionein concentration was evaluated in healthy subjects and in patients with several types of liver disorders. Plasma metallothionein concentrations in controls varied between 2.4 and 4.8 ng/ml. Patients with disorders associated with increased liver copper concentrations (i.e., primary biliary cirrhosis and primary sclerosing cholangitis) had significantly (both p <0.002) elevated plasma metallothionein concentrations (range = 1.8 to 52.2 ng/ml), and a considerable number of these were above the maximum control level (21 of 41 patients). In contrast, patients with liver disorders not associated with increased liver copper concentrations (alcoholic and cryptogenic cirrhosis, and acute viral and chronic active hepatitis) generally had normal plasma metallothionein concentrations and only a few were above the maximum control level (11 of 64 patients, maximum = 8.8 ng/ml). The metallothionein concentrations in plasma samples from patients in stage I or II primary biliary cirrhosis were within or slightly above the control range, whereas most patients in stage III had elevated levels (p <0.002), and almost all patients in stage IV had clearly elevated (p < 0.0001) concentrations. In primary biliary cirrhosis the plasma metallothionein concentration tended to increase during the evolution of the disorder, and the concentration correlated significantly with the serum total bilirubin concentration.

In conclusion, the plasma metallothionein concentration is significantly elevated in patients with primary biliary cirrhosis and in patients with primary sclerosing cholangitis. Although related to the histological stage of primary biliary cirrhosis, the measurement of plasma metallothionein concentrations contributes little to the diagnosis or the evaluation of the severity of these disorders. In patients with liver disorders not associated with increased liver copper concentrations the plasma metallothionein concentration is generally normal. (Hepatology 1991;14:1008-1012.)
Large-volume paracentesis and intravenous saline: Effects on the renin-angiotensin system
Authors: Cabrera Juan , Inglada Luis , Quintero Enrique , Jimenez Wladymiro , Losada Antonio , Mayor Juan , Guerra Cayetano
Published: March 2007

Fourteen cirrhotic patients with tense ascites were treated with total paracentesis and intravenous isotonic saline infusion. Standard liver and kidney function tests, plasma renin activity and aldosterone concentration were measured before, at 48 hrs and at 7 days after total paracentesis. The volume of ascites removed was 7.7 ± 5.6 1 (mean ± S.E.M.). None of the treated patients had clinical complications or significant alterations in liver or kidney function test results. Paracentesis and intravenous isotonic saline infusion were not associated with significant changes in mean plasma renin activity or plasma aldosterone concentration. These results suggest that this therapeutic procedure could be a safe and cost-effective alternative treatment of tense ascites in patients with cirrhosis. (Hepatology 1991;14:1025-1028.)
Predictors of one-year pretransplant survival in patients with cirrhosis
Authors: Oellerich Michael , Burdelski Martin , Lautz Hans-Ulrich , Binder Lutz , Pichlmayr Rudolf
Published: March 2007

The aim of this prospective study was to examine the usefulness of flow-dependent dynamic liver function tests and conventional methods of evaluating liver function as predictors of pretransplant survival in patients with advanced cirrhosis. Patients who underwent orthotopic liver transplantation within the follow-up period of 365 days were excluded. One hundred one patients with histologically confirmed cirrhosis were studied. Fifty-eight patients had post-hepatitic cirrhosis, 13 had cryptogenic cirrhosis and 30 had biliary cirrhosis. During follow-up, 28 patients died of their liver diseases. At entry, we recorded indocyanine green half-life, monoethylglycinexylidide formation from lidocaine, bilirubin and albumin serum concentrations, activities of cholinesterase and alkaline phosphatase, prothrombin time, clinical complications of ascites and encephalopathy and the Pugh score. These variables were subjected as covariates to a stepwise survival analysis by use of the Cox proportional-hazards model. At the final step, Pugh score, monoethylglycinexylidide formation and indocyanine green half-life were found to be the only independent variables significantly related to 1-yr survival. The parallel combination of Pugh score and monoethylglycinexylidide test yielded the highest prognostic sensitivity (82%). The series approach combining either the Pugh score and indocyanine green test or the monoethylglycinexylidide and indocyanine green tests was associated with the highest specificity (96%/97%) and high predictive values of a positive result (81%/82%). These findings suggest that appropriate combinations of the studied flow-dependent dynamic liver function tests and the Pugh score could be useful in improving transplant candidate selection and the timing of transplantation. (Hepatology 1991;14:1029-1034.)
Serum levels of short-chain fatty acids in cirrhosis and hepatic coma
Authors: Clausen Mette Rye , Mortensen Per Brøbech , Bendtsen Flemming
Published: March 2007

Short-chain fatty acids cause reversible coma in animals and may contribute to the pathogenesis of the hepatic coma in humans. The concentrations of short-chain fatty acids in peripheral venous blood were significantly elevated in 15 patients with hepatic encephalopathy caused by cirrhosis (362 ± 83 μmol/L; mean ± S.E.M.) compared with 17 cirrhotic patients without encephalopathy (178 ± 57 μmol/L) and 11 normal individuals (60 ± 8 μmol/L). However, no correlation between the depth of coma and the level of short-chain fatty acids was found after repetitive measurements in the coma group. Compared with normal individuals, all short-chain fatty acids, except valerate, were elevated in patients with hepatic encephalopathy, whereas only the concentrations of isobutyrate and isovalerate were significantly elevated in cirrhotic patients without encephalopathy.

The concentrations of short-chain fatty acids in 21 nonencephalopathic cirrhotic patients who underwent catheterization were equally distributed in the aorta (187 ± 56 μmol/L), the hepatic vein (212 ± 75 μmol/L), the azygos vein (140 ± 37 μmol/L) and the renal vein (135 ± 43 μmol/L) compared with peripheral venous blood (178 ± 57 μmol/L).

This study does not support the idea that short-chain fatty acids are of major importance in the pathogenesis of hepatic coma in patients with cirrhosis. (Hepatology 1991;14:1040-1045.)
Successful treatment of acute fascioliasis with bithionol
Authors: Bacq Yannick , Besnier Jean-Marc , Duong Thanh-Hai , Pavie Guy , Metman Etienne-Henri , Choutet Patrick
Published: March 2007

Fascioliasis is the parasitic infestation of the liver and biliary tract related to Fasciola hepatica. Bithionol is proposed as the treatment of choice for human fascioliasis without major side effects. However, the efficacy of bithionol has been evaluated in chronic but not in acute fascioliasis. In this study we report on the success of treatment with bithionol in 10 patients with fascioliasis, 8 having acute fascioliasis. The criteria for the diagnosis of fascioliasis were hypereosinophilia, positive immunoelectrophoresis and indirect hemagglutination. Bithionol was given orally to hospitalized patients at the daily dose of 25 mg/kg body wt for 10 days. Three patients with acute fascioliasis received a second course of bithionol 2 or 3 mo after the first because of the recurrence of diarrhea and fatigue in one patient and persistent hypereosinophilia in two patients. All patients were cured. The follow-up period after the first course of treatment was between 16 and 47 mo. No major side effects were observed. We conclude that bithionol is the drug of choice for both acute and chronic fascioliasis. Moreover, its oral administration may allow treatment of fascioliasis in outpatients who do not have serious symptoms. (Hepatology 1991;14:1066-1069.)
Insulin and glucagon infusion in acute alcoholic hepatitis: A prospective randomized controlled trial
Authors: Bird George , Lau Johnson Y. N. , Koskinas John , Wicks Claire , Williams Roger
Published: March 2007

In a randomized, controlled trial to investigate the possible benefit of insulin and glucagon therapy in severe acute alcoholic hepatitis, 86 patients were randomized to receive 30 U insulin and 3 mg glucagon in 250 ml 5% dextrose over 12 hr each day for 3 wk or a similar regime of identical placebo. No significant differences were seen in patients’ clinical characteristics and disease severity in the treated and placebo groups.

Of the 43 patients receiving insulin and glucagon, 15 (35%) died within 4 wk of randomization, compared with 14 deaths (33%) in the control patients (p = not significant). When the patients surviving the first 4 wk were examined there were five more deaths in the treatment group, compared with one death in the control group at 6-mo follow-up (p = not significant). No significant differences in the frequency of short-term or long-term complications of alcoholic liver disease or relapse to alcohol were seen when the two groups were compared, although hypoglycemia was seen in six patients during infusion of insulin and glucagon. Similarly, no significant differences were seen in the improvement in clinical or biochemical features at 4 wk and at 6 mo in survivors when the insulin and glucagon-treated patients were compared with patients in the placebo group.

This study does not confirm previous reports that insulin and glucagon infusion improves the outcome of severe acute alcoholic hepatitis. (Hepatology 1991;14:1097-1101.)
Immunohistochemical demonstration of pancreatic α-amylase and trypsin in intrahepatic bile ducts and peribiliary glands
Authors: Terada Tadashi , Nakanuma Yasuni
Published: March 2007

Epithelia of intrahepatic bile ducts and peribiliary glands were immunohistochemically examined for pancreatic α-amylase and trypsin in 54 normal autopsied livers. α-Amylase was evaluated with a polyclonal antibody, and trypsin was assayed with both polyclonal and monoclonal antibodies. α-Amylase was observed in large ducts, septal ducts and peribiliary glands in most livers and was seen in interlobular ducts in seven (13%) livers. Trypsin immunoreactivity with the polyclonal antibody was observed in peribiliary glands in 21 (39%) livers; it was absent in intrahepatic bile ducts in all but one liver. Trypsin immunoreactivity with the monoclonal antibody was present in large ducts, septal ducts and peribiliary glands in about 70% of the livers and was seen in interlobular ducts in two (4%) livers. Bile ductules were always negative for the two antigens. Some epithelia of peribiliary glands positive for both α-amylase and trypsin histologically resembled pancreatic acinar cells. α-Amylase and trypsin immunoreactivities of intrahepatic biliary epithelia and pancreatic aninar cells were eliminated by absorption of primary antibodies by α-amylase or trypsin, suggesting the specificities of the immunoreactivities. These data suggest that epithelia of intrahepatic large ducts, septal ducts and peribiliary glands contain pancreatic α-amylase in most livers and that they contain trypsin in about 70% of livers. α-Amylase and trypsin may be secreted into intrahepatic bile duct lumens, thereby exerting important effects on the physiology of the intrahepatic biliary tree and hepatic bile. (Hepatology 1991;14:1129-1135.)
Significance and specificity of antibodies to neutrophils detected by western blotting for the serological diagnosis of primary sclerosing cholangitis
Authors: Klein Reinhild , Eisenburg Joseph , Weber Paul , Seibold Frank , Berg Peter A.
Published: March 2007

Antibodies against neutrophils have been detected in sera from patients with primary sclerosing cholangitis and inflammatory bowel diseases either by immunofluorescence or by enzyme-linked immunosorbent assay. To assess primary sclerosing cholangitis-specific antibodies, we examined sera from 30 patients with clinically and morphologically well-established primary sclerosing cholangitis by Western blotting against neutrophils and compared these results with those obtained by testing sera from patients with inflammatory bowel diseases.

By Western blot using sonified neutrophils, 24 (80%) of 30 primary sclerosing cholangitis sera were positive. Five antigenic determinants at 95, 60, 55, 40 and 30 kD were visualized. Twenty-eight of the primary sclerosing cholangitis sera also showed the characteristic perinuclear fluorescence pattern by immunofluorescence on neutrophils. Thus a serological diagnosis of primary sclerosing cholangitis could be made in 80% of patients based on these two methods. In contrast, only 9% of 23 patients with ulcerative colitis and 10% of 60 patients with Crohn’s disease were positive by Western blot, and these patients also showed positive perinuclear fluorescence pattern by immunofluorescence, suggesting an overlap between inflammatory bowel diseases and primary sclerosing cholangitis. Although some patients with classical primary biliary cirrhosis and autoimmune chronic active hepatitis had antibodies against primary sclerosing cholangitis epitopes, none of the patients with obstructive bile duct disorders, collagen diseases, Wegener’s granulomatosis or other hepatic and nonhepatic disorders were positive by Western blot, indicating the specificity of these five primary sclerosing cholangitis-related neutrophilic epitopes. (Hepatology 1991;14:114-1152.)
Phenobarbital does not increase early labeling of bilirubin from 4-[14C]-δ-aminolevulinic acid in man and rat
Authors: Okuda Hiroaki , Tavoloni Nicola , Blaschke Terrence F. , Kiang Chi-Li , Jones Mary Jane T. , Waggoner Jeanne G. , Sardana Mohinder K. , Sassa Shigeru , Shrager Richard I. , Berk Paul D.
Published: March 2007

δ-Aminolevulinic acid-4-[14C] and [3H]-bilirubin were administered intravenously to five patients with Gilbert’s syndrome and four healthy control subjects on two occasions: before and on days 10 through 14 of a course of phenobarbital (2.5 mg/kg/day). The resulting curves of [3H]-bilirubin and [14C]-bilirubin in plasma were analyzed by computer to determine a number of parameters of physiological interest. As expected, phenobarbital produced a highly significant fall in the plasma concentration of unconjugated bilirubin as a result of a significant increase in hepatic bilirubin clearance in all subjects; plasma bilirubin turnover was unaltered. Surprisingly, the drug produced no change in the incorporation of [14C]-α-aminolevulinic acid into [14C]-early labeled bilirubin. To explain this unexpected finding, the effects of phenobarbital (75 mg/kg/day for 6 days) on incorporation of [14C]-δ-aminolevulinic acid and 2-[14C]-glycine into [14C]-early labeled bilirubin and on the activity of the enzyme δ-aminolevulinic acid synthase were studied in nonfasted, adult, male Sprague-Dawley rats. At the dose and duration of treatment used, phenobarbital administration increased total hepatic δ-aminolevulinic acid synthase activity and produced a significant increase of 70% in the incorporation of [14C]-glycine into early labeled bilirubin. By contrast, no increase in the incorporation of [14C]-δ-aminolevulinic acid into early labeled bilirubin was observed. These data suggest that δ-aminolevulinic acid is an inappropriate precursor for studies of the rate of heme biosynthesis, presumably because it bypasses δ-aminolevulinic acid synthase, the physiological rate-limiting enzyme in the heme biosynthetic pathway. (Hepatology 1991;14:1153-1160.)
Serum lysyl oxidase activity in chronic liver disease in comparison with serum levels of prolyl hydroxylase and laminin
Authors: Murawaki Yoshikazu , Kusakabe Yuka , Hirayama Chisato
Published: March 2007

Lysyl oxidase was partially purified from serum by a diethylaminoethyl batch procedure in the presence of 6 mom/L urea and dialyzed against 3 mol/L KSCN. Using this method, we determined serum lysyl oxidase activity in 52 patients with liver disease and in 14 healthy controls, and we examined usefulness of serum lysyl oxidase in assessing liver fibrogenesis. For this purpose, serum lyayl oxidase activity in chronic liver disease was compared with serum levels of prolyl hydroxylase and laminin P1. As compared with controls, serum lysyl oxidase activity increased 1.6-fold in chronic persistent hepatitis, 4.4-fold in chronic active hepatitis and 11.8-fold in cirrhosis, indicating an increase in concert with the development of liver fibrosis. In hepatocellular carcinoma, the serum activity, although signiscantly increased, was lower than that in cirrhosis. Serum prolyl hydroxylase was significantly increased in chronic active hepatitis, in liver cirrhosis and in hepatocellular carcinoma Serum laminin P1 was significantly increased in chronic active hepatitis, in cirrhosis and in hepatocellular carcinoma Serum lysyl oxidase activity did not correlate significantly with serum levels of prolyl hydroxylase and laminin P1 in any subject or in any subgroup. The magnitude of the increase and the abnormal percentage of serum lysyl oxidase activity were larger than those for serum prolyl hydroxylase and laminin P1. These results suggest that serum lysyl osidase activity is a more sensitive indicator of liver fibrosis than serum prolyl hydroxylase and laminin P1. (HEPATOLOGY 1991;14:1167-1173.)
Iron and uroporphyrin in hepatocytes of inbred mice in experimental porphyria: A biochemical and morphological study
Authors: Siersema Peter D. , van Helvoirt René P. , Ketelaars Diane A. M. , Cleton Maud I. , de Bruijn Wim C. , Wilson Paul J.H. , van Eijk Henk G.
Published: March 2007

Hexachlorobenzene-induced porphyria is iron dependent and characterized by the decreased activity of uroporphyrinogen decarboxylase and the accumulation of porphyrins in the liver. To examine the relationship between iron and porphyrins in liver tissue, we performed a biochemical and morphological (histological, ultrastructural and morphometrical) study in the livers of C57BL/10 mice. Mice were treated with hexachlorobenzene, iron dextran or the combination of hexachlorobenzene and iron dextran. An accumulation of porphyrins and an increased total iron content were found not only in the livers of mice treated with hexachlorobenzene and iron dextran but also in mice treated with iron dextran alone. In contrast, the amount of porphyrins was only slightly increased in the livers of mice treated with hexachlorobenzene alone. Needle-like structures, representing uroporphyrin crystals, were observed, histologically and ultrastructurally, in hepatocytes of mice treated with hexachlorobenzene and iron dextran and with iron dextran alone. Uroporphyrin crystals and ferritin iron were found in the same hepatocyte. A single uroporphyrin crystal, surrounded by ferritin iron, was observed in a hepatocyte of a mouse treated with hexachlorobenzene alone. Both in the livers of mice treated with hexachlorobenzene and iron dextran and in the livers of mice treated with iron dextran alone, morphometrical analysis showed that an increased area fraction of uroporphyrin crystals was associated with an increased area fraction of ferritin iron in hepatocytes.

Conclusions: In C57BL/10 mice, experimental porphyria can be induced by iron overload alone; uroporphyrin crystals and ferritin iron are located in the same hepatocyte; and the morphological co-occurrence of uroporphyrin crystals and ferritin iron in hepatocytes suggests a role for iron (as ferritin) in the pathogenesis of porphyria. (Hepatology 1991;14:1179-1188.)
Age-related changes in total protein and collagen metabolism in rat liver
Authors: Mays Peter K. , McAnulty Robin , Laurent Geoffrey J.
Published: March 2007

Liver collagen levels are determined by a balance between synthesis and degradation, processes known to have rapid rates in growing animals. We report age-related changes in liver collagen synthesis and degradation rates, as well as protein synthesis rates, in rats at five ages from 1 to 24 mo. Fractional collagen synthesis rates were determined after injection of [14C]proline with a flooding dose of unlabeled proline and its incorporation as hydroxy-[14C]proline into proteins. Fractional protein synthesis rates were based on the uptake of [14C]proline into proteins. Fractional collagen degradation rates were calculated from the difference between collagen fractional synthesis and deposition rates. Fractional rates of collagen synthesis were similar between 1 mo (23.0% ± 4.6%/day) and 24 mo (19.6% ± 3.4%/day) of age. Collagen deposition into the extracellular matrix was extremely low at every age studied; therefore degradation pathways accounted for the bulk of the collagen synthesized. The mean fractional synthesis rate for the total protein pool was unaltered between 1 mo (105.0% ± 7.2%/day) and 15 mo (89.9% ± 6.0%/day) of age, after which it increased to 234.9% ± 33.0%/day (p < 0.05) by 24 mo of age. These results indicate that liver collagen and total protein synthesis rates were maintained at relatively high levels during development and maturity but that protein synthesis rates were highest in senescent animals. (Hepatology 1991:14:1224-1229.)
Enhanced biliary iron excretion with amphiphilic diethylenetriaminepentaacetic acid
Authors: Adams Paul C. , Lin Edward , Barber Kathryn R. , Grant Chris W. M.
Published: March 2007

The elimination of toxic metal ions metabolically accumulated by patients remains a difficult clinical problem and a target of drug development. DTPA (diethylenetriaminepentaacetic acid) is a hydrophilic chelating agent with high affinity for divalent and trivalent metal ions including iron but with a limited ability to cross cell membranes for access to iron stores. In this study we have synthesized an amphiphilic form of this chelator-DTPA covalently linked to the phospholipid phosphatidylethanolamine (PE)-to produce a chelator that incorporates completely and stably into liposome membranes for efficient delivery to the liver and reticuloendothelial system. Biliary and urinary excretion of iron were studied in iron-loaded rats (n = 15) in association with a 2-hr intravenous infusion of sonicated liposomes of 1:1 amphiphilic phosphatidylethanolamine-DTPA/egg phosphatidylcholine (L-PE-DTPA) and compared with excretion obtained using equivalent amounts of watersoluble DTPA (alone or mixed with egg phosphatidylcholine liposomes [L-DTPA] as controls). For a 6-hr period, the administration of L-PE-DTPA resulted in approximately a 20-fold increase in biliary iron excretion (480 ± 160 μg/6 hr, mean ± S.D.) compared with that seen with DTPA (21.2 ± 4.0μg/6 hr) and L-DTPA (23.1 ± 5.0 μg/6 hr) (p < 0.05, analysis of variance). Urinary iron excretion was significantly decreased with L-PE-DTPA (41.5 ± 38 μg/6hr) compared with DTPA (154 ± 110 μg/6 hr) and L-DTPA (86 ± 17 μg/6 hr) (p < 0.05). Combined biliary and urinary excretion of iron was three to four times greater with L-PE-DTPA. This study suggests that the use of a highly water-insoluble amphiphilic form of DTPA as sonicated liposomes altered the route and extent of delivery of the chelator to the iron stores, resulting in a significant increase in biliary iron excretion. (Hepatology 1991;14:1230-1234.)
Cardiovascular responses to serotonin in experimental liver disease
Authors: Jacob Giris , Bishara Bashara , Lee Samuel S. , Hilzenart Nir , Bomzon Arieh
Published: March 2007

Recent evidence suggests that serotonergic mechanisms within the cardiovascular system are activated and may be important in the development of the hyperkinetic circulation and the maintenance of portal hypertension in cirrhotic patients. The in vivo pressor and positive chronotropic response together with the in vitro contractile responses of aortic rings and portal veins to serotonin were studied in three different rat models of cirrhosis, portal hypertension and jaundice: the portal vein-ligated rat, the carbon tetrachloride-induced cirrhotic rat, the chronic bile duct-ligated cirrhotic rat and the 3-day noncirrhotic, nonportal hypertensive-jaundiced rat. In addition, the activity of the enzyme monoamine oxidase type A was determined in lung homogenates prepared from the four groups of sham and treated animals. In the four different groups of sham-treated or operated pithed rats, serotonin caused a dose-dependent increase in mean arterial blood pressure without any effect on the heart rate. The pressor responses to serotonin in the three models of portal hypertension were significantly attenuated from their respective sham group. In the 3-day noncirrhotic, nonportal hypertensive-jaundiced rats, the pressor response was no different than that seen in the sham-operated rats. No evidence of consistent potentiated or blunted in vitro reactivity to serotonin of arterial rings and portal veins from the four groups of rats was seen. Portal hypertension, cirrhosis and hyperbilirubinemia had no effect on the activity of monoamine oxidase type A. These data demonstrate that portal hypertension is associated with an attenuated pressor response to serotonin. The mechanism of this attenuated response is not linked to altered in vitro reactivity of arterial rings to serotonin or increases in the activity of pulmonary monoamine oxidase. Because the in vitro reactivity of portal veins from portal hypertensive rats was not hypersensitive to serotonin, our data are consistent with the proposal that serotonin2-antagonists exert their portal hypotensive action by lowering resistance in the mesenteric and collateral circulation of cirrhotic patients. (Hepatology 1991;14:1235-1242).
Role of microtubuli in secretion of very-low-density lipoprotein in isolated rat hepatocytes: Early effects of thiol reagents
Authors: Nagelkerke Fred J. , van de Water Bob , Twiss Irene M. , Zoetewey Paul J. , de Bont Hans J. G. M. , Dogterom Peter , Mulder Gerard J.
Published: March 2007

The secretion of very-low-density lipoprotein from hepatocytes proceeds through the microtubules. In this study, the role of glutathione in the maintenance of intact microtubules and the secretion of very-low-density lipoprotein has been investigated. When rat hepatocytes were incubated with reagents that deplete glutathione (e.g., diethylmaleate, α-bromoisovalerylurea or allyl alcohol) or reacted directly with protein thiols (disulfiram), the secretion of very-low-density lipoprotein by the cells was inhibited and the microtubules were severely damaged as visualized by immunofluorescence staining. Both events occurred within 30 min; long before, an effect on the energy status of the cells became evident. The observed inhibition of the secretion therefore seems due to an effect of the toxicants on the microtubules. For α-bromoisovalerylurea, diethylmaleic acid and allyl alcohol, it may be related to glutathione depletion; preincubation of the hepatocytes with N-acetyl-l-cysteine reduced the decrease of glutathione by α-bromoisovalerylurea and allyl alcohol (but not of diethylmaleic acid) and almost completely prevented the inhibition of very-low-density lipoprotein secretion and microtubule damage. Depletion of glutathione may result in modification of a small group of essential free protein thiols. Disulfiram did not deplete glutathione, and N-acetyl-l-cysteine could not prevent the effects of disulfiram on microtubules. The binding to protein thiols of radiolabeled disulfiram, which binds to microtubules in vitro, was determined. At 0.2 mmol/L disulfiram, only 3% of total cellular protein thiols were conjugated, but secretion of very-low-density lipoprotein was already inhibited by 25%, and microtubules were severely affected. We propose that modification of a small fraction of cellular protein thiols results in the loss of microtubular ultrastructure and thereby leads to inhibition of very-low-density lipoprotein secretion. (Hepatology 1991;14:1259-1268.)
Physiological responses to increased dietary cholesterol: The case of the egg man
Authors: Gumucio Jorge J. , Jones Michael P. , Heuman Douglas M.
Published: March 2007

An 88-yr-old man carrying the tentative diagnosis of Alzheimer’s disease had a documented history of ingesting 20 to 30 eggs/day for approximately 15 yr. His psychiatrist characterized his eating habits as a compulsive disorder:“Eating these eggs ruins my life, but I can’t help it.” He is in excellent health. His weight is constant at 82 to 86 kg (height 1.87 m). Apparently, he had a mild episode of angina. Otherwise, no significant history of stroke, heart disease or gallbladder disease was seen. His serum cholesterol levels have ranged between 150 and 200 mg/dl.