Hepatology

doi: 10.1002/hep.1840060501pmid: N/A

Gluud, Christian; Gluud, Christian

doi: 10.1002/hep.1840060502pmid: N/A

A double‐blind, placebo‐controlled multicenter trial was conducted to determine the efficacy of oral testosterone treatment (200 mg three times daily) in men with alcoholic cirrhosis. By skewed randomization (3:2), 134 patients received testosterone and 87 placebo. Patients were followed from 8 to 62 months (median = 28 months). In the testosterone group, 33 patients died (25%; 95% confidence limits = 18 to 33%) as compared to 18 (21%; 95% confidence limits = 13 to 31%) in the placebo group. Taking age and significant prognostic variables into consideration, this corresponds with a relative mortality risk of 1.17 (95% confidence limits = 0.65 to 2.15) in the testosterone group vs. the placebo group. Testosterone treatment did not significantly affect liver biochemistry, prevalence of complications to cirrhosis or causes of death. Patients treated with testosterone developed significantly (p < 0.05) higher serum testosterone and blood hemoglobin concentrations and significantly (p < 0.05) lower plasma IgM concentrations as compared to the placebo group. The prevalence of gynecomastia decreased significantly (p < 0.05) in the testosterone group as compared to the placebo group. We conclude that oral testosterone treatment has no beneficial effect on survival and liver biochemistry in men with alcoholic cirrhosis, and adverse effects cannot be excluded.

Tsukamoto, Hidekazu; Towner, Sally J.; Clofalo, Lefran M.; French, Samuel W.

doi: 10.1002/hep.1840060503pmid: 3758935

Severe hepatic steatosis and focal necrosis of hepatocytes have previously been induced in rats by continuous intragastric infusion of ethanol and a diet containing only 5% fat as a per cent of total calories as reported by us previously. Since the ethanol diet fed ad libitum with such a low level of fat has previously failed to produce alcoholic fatty liver, continuously high blood alcohol levels achieved in this model appeared to be key in induction of the progressive pathologic lesions in the liver. In the current study, effects of increased fat intake on alcohol‐induced liver injury were investigated. Seventeen pairs of male Wistar rats were implanted with gastrostomy cannulas and infused with a liquid diet containing 25% of total calories as fat plus ethanol or isocaloric dextrose. Ethanol intake was progressively increased from 32% up to 47% of total calories to maintain sustained intoxication for 30 to 120 days. Light and electron microscopic examination of the liver revealed moderate to severe fatty infiltration in all of the ethanolfed rats, of which 14 had spotty or zonal necrosis in the centrilobular areas accompanied by polymorphonuclear and mononuclear cell infiltration. In addition, fibrosis was observed in association with the necrotic lesions or with large‐droplet steatosis. Reticulin and trichrome stains clearly demonstrated fine fibrosis, including perivenular fibrosis as well as septum formation progressing to bridging fibrosis. Furthermore, increased numbers of Ito cells and myofibroblasts were observed in the perivenular fibrotic areas. These results demonstrate striking potentiation of alcohol‐induced liver injury by the increased fat intake or by the concomitant decrease in carbohydrate intake. This model, which represents the first rat model to achieve such advanced experimental alcohol‐induced liver injury, possesses the obvious potential for further use in studies of ethanol‐nutrient interactions in the pathogenesis of alcohol‐induced liver injury.

Wilson, Jeremy S.; Korsten, Mark A.; Lieber, Charles S.

doi: 10.1002/hep.1840060504pmid: 3530943

This investigation was performed to examine the combined effects of protein deficiency and chronic ethanol consumption on ethanol clearance and hepatic ethanol metabolism of the rat. Protein deficiency alone was associated with reduced ethanol clearance and decreased activity of hepatic alcohol dehydrogenase and the microsomal ethanol‐oxidizing system. However, when ethanol (as 36% of energy) was administered concurrently with protein‐deficient diets, accelerated ethanol clearance and increased microsomal oxidation of ethanol was observed. Furthermore, in protein‐deficient animals fed ethanol, liver alcohol dehydrogenase levels were less decreased when compared with values observed in animals fed protein‐deficient diets without ethanol, and this effect was associated with markedly reduced serum testosterone levels in the former group.

Steffan, Anne‐Marie; Gendrault, Jean‐Louis; McCuskey, Robert S.; McCuskey, Patricia A.; Kirn, André

doi: 10.1002/hep.1840060505pmid: 3758936

Impairment of the phagocytic capacities of Kupffer cells, as is found in Frog Virus 3 hepatitis of mice, allows the endothelial liver cells to take up intravenously inoculated latex particles of 1.0 μm diameter. In vitro experiments with cultivated endothelial cells isolated by collagenase perfusion of the liver and purified by centrifugal elutriation demonstrate that uptake occurs via a typical mechanism of phagocytosis involving pseudopodia. Ingestion of latex is inhibited by incubation of the cells at 4°C and by treatment with cytochalasin B, whereas colchicine has no effect. These results demonstrate that: (i) the Kupffer cells are not the only cells of the hepatic sinusoid capable of phagocytosis; and (ii) under conditions where the phagocytosis in Kupffer cells is impaired, the endothelial cells may participate in the clearance of large particles from the blood.

Schiff, J. Michael; Huling, Sandra L.; Jones, Albert L.

doi: 10.1002/hep.1840060506pmid: 3758937

The degradation of asialoglycoproteins in hepatocytes has been well described in several animal models, but no direct evidence has yet been obtained for asialoglycoprotein processing in the primate liver. A double radiolabeling strategy was employed in the experiments described in this paper to evaluate the fate of asialoorosomucoid in the squirrel monkey. Intravenously injected asialoorosomucoid was taken up by the liver with a half‐time of 1 min. Electron microscopic autoradiography showed progression of asialoorosomucoid from the hepatocyte plasma membrane through vesicles to multivesicular bodies and then to secondary lysosomes near the Golgi‐rich area of the cell. Over 75% of the grains initially associated with clear endocytic compartments after injection had moved to these later organelles within 20 min. Following degradation of asialoorosomucoid labeled with the Bolton and Hunter reagent, radiocatabolites were secreted into bile, peaking ∼47 min after injection. We also found that 7 to 8% of the injected protein entered an alternative pathway which led to resecretion of the ligand at the bile canaliculus. This was considerably more than in rats (1 to 3%), but roughly comparable to the amount in guinea pigs (10 to 17%). Intact asialoorosomucoid peaked in monkey bile∼27 min after injection and was 3 to 4 times more concentrated than the initial plasma concentration, indicating receptor‐mediated transport. Gel filtration chromatography and polyacrylamide gel analysis of the secreted protein indicated that it had arrived in bile unaltered. Since <1% of the autoradiographic grains were localized to nonparenchymal cells, the hepatocyte was identified as the cell type simultaneously responsible for both pathways. We propose that missorting of some of the asialoglycoprotein to bile reflects diffusion within intracellular sorting compartments to areas primarily dedicated to the processing of unrelated ligands, such as those newly synthesized for biliary secretion.

Cleton, Maud I.; Frenkel, Erik J.; de Bruijn, Willem C.; Marx, Joannes J. M.

doi: 10.1002/hep.1840060507pmid: 3758938

The amounts of phosphorus and iron in various isolated ferritin preparations were investigated by: (i) chemical analysis on ferritin samples and (ii) electron probe X‐ray microanalysis on ferritin particles from the same preparations. A high correlation was found between iron to phosphorus ratios obtained by both methods. Further investigation by electron probe X‐ray microanalysis on lysosomes of hepatic cells of patients with idiopathic and secondary hemochromatosis revealed lysosomal iron to phosphorus ratios which were very similar in all parenchymal cells but different from ratios obtained in Kupffer cells. Lysosomal iron to phosphorus ratios in hepatocytes did not change after intensive phlebotomy treatment. It is postulated therefore that, during phlebotomy, iron and phosphorus are concomitantly lost from the hepatic lysosomes.

Trinder, Deborah; Morgan, Evan; Baker, Erica

doi: 10.1002/hep.1840060508pmid: 3758939

The mechanisms of iron accumulation by cultured hepatocytes isolated from fetal rat liver (19 days gestation) were investigated using rat transferrin labeled with 125I and 59Fe. The rates of iron and transferrin internalization by the cells were measured by incubating the hepatocytes with the labeled transferrin at 37°C followed by treatment with pronase at 4°C to remove surface‐bound transferrin and iron. Iron internalization increased linearly with time. Approximately 65% of the internalized iron was incorporated into ferritin. In contrast to iron, the rate of transferrin internalization was biphasic, with a rapid phase during the first 10 to 15 min and a second slower phase which becomes more apparent after that time. Iron and transferrin internalization were temperature‐dependent. Chase experiments showed that the internalized transferrin donated all of its iron to the cell and was then released in a biphasic manner which was dependent on the time of preincubation with radiolabeled transferrin. These experiments showed that iron uptake occurs by at least three processes. The first mechanism involves the specific receptor‐mediated endocytosis of transferrin. Each cell has an average of 7.8 ± 1.0×105 (mean ± SE, n = 5) transferrin binding sites with an apparent association constant of 2.0 ± 0.4×106 M−1. The second process is nonsaturable up to a transferrin concentration of at least 6 μM but like the specific process, also leads to accumulation of iron in excess of transferrin. It involves the endocytosis of transferrin mediated by 4.2×106 transferrin binding sites which have a K′a value of 2.6×105 M−1. Third, fluid‐phase endocytosis of transferrin occurs at a rate of 0.13×104 molecules per cell per min at 1.25 μM transferrin concentration. This rate is not sufficient to account for the additional iron accumulated by the cells by the second process.

Shaver, William A.; Bhatt, Harshika; Combes, Burton

doi: 10.1002/hep.1840060509pmid: 3758940

Low values for serum alkaline phosphatase activity were observed early in the course of two patients with Wilson's disease presenting with the combination of severe liver disease and Coombs' negative acute hemolytic anemia. A review of other cases of Wilson's disease revealed that 11 of 12 patients presenting with hemolytic anemia had values for serum alkaline phosphatase less than their respective sex‐ and age‐adjusted mean values; in eight, serum alkaline phosphatase activity was less than the lower value for the normal range of the test. Low values for serum alkaline phosphatase were much less common in Wilson's disease patients with more chronic forms of presentation. Copper added in high concentration to serum in vitro did not have an important effect on serum alkaline phosphatase activity. The mechanism responsible for the decrease in serum alkaline phosphatase activity in patients is uncertain.

Gaulard, Philippe; Zafrani, Elie Serge; Mavier, Philippe; Rocha, Francisco Dario; Farcet, Jean‐Pierre; Divine, Marine; Haioun, Corinne; Pinaudeau, Yvon

doi: 10.1002/hep.1840060510pmid: 3530944

Three cases of a peculiar form of peripheral T‐cell lymphoma presenting as predominant hepatic disease with splenomegaly are reported. The three patients had marked liver enlargement without lymphadenopathy; white blood cell count was normal, and modifications of hepatic tests were mild. In the three cases, the diagnosis of the lymphoma was mainly based on the results of hepatic morphological changes. Liver involvement was histologically characterized by a predominantly sinusoidal infiltration by tumor cells in the three cases, associated with perisinusoidal fibrosis in two of them; portal infiltration was noted in two patients. Immunopathological study showed that tumor cells were T‐lymphoid cells that were different from normal T‐lymphocytes by the lack of expression of one T‐cell membrane antigen, i.e., Leu‐1. These findings suggest that a distinct clinical, pathological and immunopathological entity might be individualized within the large group of T‐cell lymphomas.