bluuebunny/crossref_metadata_2025 huggingface dataset:
Partial purification of rat hepatic stimulator substance and characterization of its action on hepatoma cells and normal hepatocytes
Authors: Fleig Wolfgang E. , Hoss Gerhard
Published: March 2007

The active principle of a cytosol extract from weanling rat liver representing a putative liver-specific growth factor was partially purified and characterized. “Hepatic stimulator substance” was extracted from the livers of 40- to 60-gm male rats by heat treatment of a homogenate in 35% (w/v) phosphate-buffered saline and subsequent ultracentrifugation. This “heat supernatant” and fractions derived from the subsequent purification steps were tested for growth stimulatory activity in two rat hepatoma cell lines. The undifferentiated, fibroblas-toid-like HTC hepatoma cells did not respond to crude hepatic stimulator substance or any of the partially purified preparations. In contrast, MH1C1 cells, which display some differentiated hepatic functions and epithelial morphology, reacted to hepatic stimulator substance and the purified fractions with a dose-dependent increase of their growth rate in serum-free culture. Although insulin, glucagon and epidermal growth factor showed only a minor effect on MH1C1 cell growth on their own, they were active as permissive or potentiating factors for the expression of the maximal effect of hepatic stimulator substance. Similarly, normal adult rat hepatocytes were only sensitive to hepatic stimulator substance when cultured in the simultaneous presence of epidermal growth factor. Under such conditions, hepatic stimulator substance stimulated hepatocyte entry into the S-phase of the cell cycle 3-fold compared to epidermal growth factor alone. Hepatic stimulator substance did not affect growth of human skin fibroblasts and of the rat intestinal crypt cell line IEC-6. When compared to the supernatant obtained after a 65°C heat step, heating to 95°C increased the biological activity of the extract 2- to 3- fold. MH1C1 cell growth with 50 μg protein per ml of the 95°C heat supernatant in serum-free medium was identical to the growth rate obtained with 10% fetal bovine serum. Precipitation with ethanol reduced the growth-stimulatory activity of hepatic stimulator substance by 60%. The biological activity was concentrated 10- and 100-fold by conventional and high-performance gel filtration, respectively. The biological activity was non-dialyzable and was destroyed by protease treatment. In sodium dodecyl slfate-polyacrylamide gel electrophoresis, the partially purified material showed three bands between M.W. 15,300 and 20,000.
Bone disease in chronic childhood cholestasis. I. vitamin D absorption and metabolism†
Authors: Heubi James E. , Hollis Bruce W. , Specker Bonny , Tsang Reginald C.
Published: March 2007

Metabolic bone disease is common in children and adults with chronic cholestasis. We evaluated baseline vitamin D (vitamin D2 and D3), 25-OH vitamin D2 and D3, 1,25(OH)2 vitamin D, vitamin D-binding protein, bone mineral content and dietary mineral content in six children (mean age: 12.1 years) with cholestasis since infancy. Absorption of 25-OH vitamin D3 and vitamin D2 was evaluated by measuring serial serum concentrations after a test dose. Bone mineral content was reduced by >2 S.D. in five of six subjects compared to age-specific controls; none had radiographic evidence of rickets but all had osteopenia. Dietary Ca and P content in the subjects was comparable to the recommended daily allowance for age-specific children. Baseline serum vitamin D2 concentrations were undetectable in all but one cholestatic subject despite oral supplementation with 2,500 to 50,000 IU per day vitamin D2. Baseline serum 25-OH vitamin D was 33.2 ± 6.0 ng per ml (mean ± S.E.) and comparable to our laboratory norms (15 to 50 ng per ml). Serum 1,25(OH)2 vitamin D and “free” 1,25(OH)2 vitamin D were both significantly (p<0.05) reduced compared to controls. A significantly blunted rise and reduced area under the absorption curve (both p<0.001) after 1,000 IU per kg vitamin D2 was found in cholestatic children (0.8 ng ± 0.5 ng per ml and 18.0 ± 14.3 ng hr per ml, respectively) compared to controls (59.5 ± 10.0 ng per ml and 1,780 ± 253 ng hr per ml, respectively). The peak change and area under the absorption curve for 25-OH vitamin D3 from baseline after 10 μg per kg 25-OH vitamin D3 was significantly reduced (both p<0.05) in cholestatic children (60.2 ± 13.8 ng per ml and 751.5 ± 189.9 ng hr per ml, respectively) compared to controls (151.6 ± 30.7 ng per ml and 2,021 ± 472 ng hr per ml, respectively). We conclude that despite severe vitamin D malabsorption, normal serum 25-OH vitamin D concentrations in cholestatic children most likely result from sunlight-stimulated endogenous vitamin D synthesis. Use of oral 25-OH vitamin D should be considered in cholestatic children because its absorption is less impaired than vitamin D.
A model of presinusoidal portal hypertension
Authors: Lefkowitch Jay H.
Published: March 2007

The etiology of idiopathic portal hypertension (IPH) is not known. To obtain clues to the pathogenesis, an attempt was made to produce a hepatic lesion similar to that in IPH by repeated injections of aggregated killed non-pathogenic E. coli directly into the portal vein. In the treated dogs, histology of the liver showed dense fibrosis in the portal tract and an aberrant vasculature around the portal area after 1 month. Portal pressure was elevated and middle-to-small-sized portal branches were decreased in number as studied by portography. These changes closely mimic those seen in human IPH. The possibility is discussed that chronic entrance of an antigen such as bacteria from the intestine to the portal venous system plays an etiologic role in IPH.
Increased tumor necrosis factor production by monocytes in alcoholic hepatitis
Authors: McClain Craig J. , Cohen Donald A.
Published: March 2007

Tumor necrosis factor is a cytokine that mediates many of the biologic actions of endotoxin. Recent studies have shown that tumor necrosis factor administration may cause liver injury and that tumor necrosis factor may mediate the lethality of the hepatotoxin galactosamine. One of the most potent inducers of tumor necrosis factor production is endotoxin. Because patients with alcoholic liver disease frequently have endotoxemia and because many of the clinical manifestations of alcoholic hepatitis are known biologic actions of tumor necrosis factor, we thought it would be important to evaluate tumor necrosis factor activity in patients with alcoholic hepatitis. Basal and lipopolysaccharide-stimulated tumor necrosis factor release from peripheral blood monocytes, a major source of tumor necrosis factor production, was determined in 16 patients with alcoholic hepatitis and 16 healthy volunteers. Eight of 16 alcoholic hepatitis patients and only two of 16 healthy volunteers had detectable spontaneous tumor necrosis factor activity (p < 0.05). After lipopolysaccharide stimulation, mean monocyte tumor necrosis factor release from alcoholic hepatitis patients was significantly increased to over twice that of healthy controls (25.3 ± 3.7 vs. 10.9 ± 2.4 units per ml, p < 0.005). We conclude that monocytes from alcoholic hepatitis patients have significantly increased spontaneous and lipopolysaccharide-stimulated tumor necrosis factor release compared to monocytes from healthy volunteers. We suggest that some of the metabolic abnormalities and possibly some of the liver injury of alcoholic hepatitis may be due to enhanced tumor necrosis factor production.
Isolation, culture and main characteristics of mouse fat-storing cells: Interaction with viruses
Authors: Chen Wan , Gendrault Jean-Louis , Steffan Anne-Marie , Jeandidier Eric , Kirn André
Published: March 2007

Fat-storing cells were isolated from 15-day-old mouse sinusoidal cell cultures (Kupffer or endothelial cells), where they had multiplied abundantly; they were then purified by a negative selection method based on the fact that they do not possess Fc receptors, as do both other types of cells.

The fat-storing cells, which could be subcultured for at least 10 passages, have the main morphological characteristics already described in vivo, in particular, the rough endoplasmic reticulum and the lipid droplets, which become less and less apparent as the number of passages increases. Subcultured fat-storing cells, almost devoid of lipid droplets and vitamin A, were able to take up retinol, as the appearance of a typical autofluorescence indicated; the number of lipid droplets increased concomitantly. Furthermore, the cultured fat-storing cells were able to internalize one-μm-sized latex beads by phagocytosis. Infection of fat-storing cells with mouse hepatitis virus 3, ectromelia or Sindbis virus led to multiplication of the virus particles. There was a direct relation between the multiplication of mouse hepatitis virus 3 in cultured fat-storing cells and the susceptibility of the animals to the virus. In the case of Sindbis virus, interferon is produced, its production being independent of the presence of vitamin A.
Urea cycle enzyme activities are normal and inducible by a high-protein diet in CCl4 cirrhosis of rats
Authors: Snodgrass Philip J.
Published: March 2007

We produced moderately severe, inactive micronodular cirrhosis in rats using CCl4 and measured the urea cycle enzyme activities in liver after feeding a 15% casein diet for 1 week and again after a 60% casein diet for 1 week. There was no deficiency of any of the five urea cycle enzymes in cirrhotic livers of rats pair-fed the 15% casein diet. Argininosuccinate synthetase and carbamyl phosphate synthetase activities were lower than in non-pair-fed controls by some baselines. All five enzymes in cirrhotic livers were induced 1.5- to 3-fold by the high-protein diet expressed as units per 100 gm of rat. The level of carbamyl phosphate synthetase activity was lower in the livers of rats pair-fed the 60% casein diet than in control livers based on wet weight, collagen-free protein and DNA, but the activities were equal expressed as units per 100 gm of rat. This example of CCl4-induced cirrhosis in the rat does not serve as a good model for human cirrhosis, in which the urea cycle enzymes are reported to be decreased in activity.
Patterns of intrahepatic recurrence after curative resection of hepatocellular carcinoma
Authors: Matsumata Takashi , Kanematsu Takashi , Takenaka Kenji , Yoshida Yasuhiro , Nishizaki Takashi , Sugimachi Keizo
Published: March 2007

Using selective angiograms of the liver, we analyzed the patterns of intrahepatic recurrence of hepatocellular carcinomas following curative surgery. In 33 patients with intraheptaic recurrences, seven patients (21%) had a recurrence near the resected hepatic stump. The remaining 26 had either a nodular recurrence in segments away from the resected margin or a widespread multinodular recurrence in the liver remnant. There was a recurrence within the first postoperative year in five of 16 patients with a nodular recurrence and in eight of 10 patients with a widespread multinodular recurrence. In these patients, particularly those with a widespread multinodular recurrence, tumor thrombi in the portal vein present before the operation and/or disseminated during operation from such an advanced main tumor seemed to be the most important and significant factor related to the early recurrence in the remnant liver. This evidence suggests that in cases of surgery for hepatocellular carcinoma it is important to establish a technique to prevent dissemination of cancer cells due to operative manipulation and also adequate adjuvant therapy rather than attempting to obtain an ample resection margin.
Hyperparathyroidism and portacaval shunt: A syndrome or a coincidence?
Authors: Urban Ernest , Dindzans Vincent J. , Van Thiel David H.
Published: March 2007

This is a report of six patients with cirrhosis of the liver in whom primary hyperparathyroidism occurred due to a solitary parathyroid adenoma 3 months to 9 years after undergoing emergency portacaval shunt for hemorrhage from esophageal varices. The presenting symptoms in all six patients were weakness and bone pain. Three patients had a bone fracture after insignificant trauma, one and probably two passed kidney stones, and a duodenal ulcer developed in two. Bone x-ray films showed generalized osteoporosis in all patients. Renal function and arterial blood pH were within normal limits in every patient. The diagnosis of primary hyperparathyroidism in each patient was based on repeated demonstrations of hypercalcemia, hypophosphatemia, and markedly elevated serum immunoreactive parathyroid hormone concentrations. In all six patients, removal of the parathyroid adenoma resulted in disappearance of symptoms; normalization of serum calcium, phosphorus, and immunoreactive parathyroid hormone levels; and, in four of the six, improvement in radiographic evidence of osteoporosis during follow-up of from 1 to 6 years. The association of cirrhosis, portacaval shunt, and primary hyperparathyroidism has not been documented previously. Our six patients with primary hyperparathyroidism constitute 3.4 percent of 174 survivors of emergency portacaval shunt in a series of 264 unselected, consecutive patients with cirrhosis and bleeding esophageal varices. Hepatic osteodystrophy is known to have occurred in only 11 of these 174 survivors. Primary hyperparathyroidism may be a more common cause of hepatic osteodystrophy than has been previously recognized, and should be considered in patients with cirrhosis in whom weakness, bone pain, and bone demineralization develop, particularly if they have a portacaval anastomosis.
The prevalence and incidence of cholecystolithiasis in patients with chronic liver diseases: A prospective study
Authors: Sheen I-Shyan , Liaw Yun-Fan
Published: March 2007

To investigate the prevalence and incidence of cholecystolithiasis in hepatitis B surface antigen-positive chronic liver diseases, a prospective study using hepatobiliary ultrasonography was conducted in 933 “healthy” persons and four groups of HBsAg-positive patients: 226 asymptomatic carriers, 73 patients with chronic active hepatitis, 77 patients with early liver cirrhosis and 124 patients with advanced liver cirrhosis. Patients with hepatocellular carcinoma or alcoholism were excluded. The prevalences of cholecystolithiasis increased along with the increasing duration and severity of chronic liver diseases, with a significant linear trend (p < 0.001). The prevalences of cholecystolithiasis in patients with liver cirrhosis (18.5% in males, 31.2% in females) were 4 to 5.5 times higher than that of the healthy population (p < 0.005). In addition, the linear trend of increasing prevalence with increasing age in the healthy population was not observed in patients with chronic liver diseases. On the other hand, five of the 69 patients with early liver cirrhosis were found to develop cholecystolithiasis during a mean follow-up period of 32 months. The calculated annual incidence of cholecystolithiasis was 2.6%. The results suggest that chronic liver disease, particularly liver cirrhosis, is a risk factor for cholecystolithiasis.
Effect of repetitive low-dose endotoxin on liver parenchymal and kupffer cell fibronectin release
Authors: Vincent Peter A. , Cho Eshin , Saba Thomas M.
Published: March 2007

Repetitive low-dose endotoxin, at a dose which will result in endotoxin tolerance, produces a marked but transient 2- to 3-day increase in plasma fibronectin. This elevation of fibronectin appears to contribute to increased hepatic Kupffer cell phagocytic function observed with repetitive low-dose endotoxin administration. Although numerous cell types synthesize fibronectin, hepatocytes are believed to be the major cell source of fibronectin in the plasma. Since Kupffer cells also synthesize fibronectin, we sought to determine the relative contribution of hepatic Kupffer cells, as compared to parenchymal cells, to the elevation of plasma fibronectin following repetitive low-dose endotoxin administration. Kupffer cells isolated from rats previously treated for 3 consecutive days with 100 μg Salmonella enteritidis endotoxin released greater (p < 0.01) amounts of fibronectin over time in culture (3, 6, 12 and 24 hr) as compared to Kupffer cells isolated from normal rats. Experiments in which fibronectin was normalized to DNA content of the cells in culture also showed similar results for fibronectin release by Kupffer cells (normal: 2.9 ± 0.5 ng per μg DNA per 24 hr; endotoxin-treated: 53.3 ± 1.3 ng per μg DNA per 24 hr). Hepatocytes from endotoxin-treated rats released less (p < 0.01) fibronectin over time than hepatocytes isolated from normal animals. As with Kupffer cells, results for fibronectin release by hepatocytes were similar when normalized to the DNA content (normal: 190.0 ± 9.4 ng per μg DNA per 24 hr; endotoxin-treated: 83.3 ± 4.2 ng per μg DNA per 24 hr). However, cultured hepatocytes did synthesize and release more fibronectin than cultured Kupffer cells whether they were isolated from normal or endotoxin-treated rats. These results suggest that Kupffer cells may contribute a greater portion of the plasma fibronectin pool following endotoxin treatment than in normal animals. Furthermore, the increased release of fibronectin by Kupffer cells observed after endotoxin is coupled with decreased hepatocyte release of fibronectin.
Biochemical effects and zonal heterogeneity of peroxisome proliferation induced by perfluorocarboxylic acids in rat liver
Authors: Just Wilhelm W. , Gorgas Karin , Hartl Franz-Ulrich , Heinemann Petra , Salzer Manfred , Schimassek Hans
Published: March 2007

Rats were treated for 5 to 14 days with perfluoroacetate, perfluorobutyrate and perfluorooctanoate. Alterations in hepatic morphology with special reference to the peroxisomal compartment were investigated by light and electron microscopy following cytochemical staining of catalase activity with the alkaline 3,3′-diaminobenzidine medium. All three compounds induced hepatomegaly and peroxisome proliferation. Perfluorobutyrate and perfluorooctanoate were found to be more active than perfluoroacetate. Perfluorooctanoate-induced peroxisome proliferation was more prevalent in centrilobular than in periportal hepatocytes. Peroxisomes in centrilobular liver cells frequently were of round shape, exhibited diameters of up to 1.5 μm and were predominantly located within smooth endoplasmic reticulumglycogen areas. In periportal cells, however, clusters of polymorphous peroxisomes ranging from 250 to 1,100 nm in diameter were observed at the periphery of smooth endoplasmic reticulum-glycogen regions. Peroxisome proliferation was accompanied by a change of peroxisomal and mitochondrial enzyme activities, in particular an increase in peroxisomal palmitoyl-CoA oxidation. Significant alterations in the concentration of peroxisomal matrix and membrane polypeptides were also noted. Within the first 2 days, perfluorooctanoate treatment exerted a strong hypolipidemic activity and both compounds perfluorooctanoate and perfluorobutyrate raised the level of hepatic free acid-soluble CoA nearly 10-fold as compared with control livers. The results suggest perfluorinated carboxylic acids to be model substances suitable to correlate biochemical and morphological parameters with the zonal heterogeneity of the peroxisomal compartment in rat liver. Due to the manifold hepatic effects, contact of humans with perfluorinated carboxylic acids or their metabolic precursors may represent a severe health risk.
Vitamin B6 repletion in cirrhosis with oral pyridoxine: Failure to improve amino acid metabolism
Authors: Henderson Michael J. , Scott Steven S. , Merrill Alfred H. , Hollins Bettye , Kutner Michael H.
Published: March 2007

This study evaluated the effect of daily oral pyridoxine supplementation in patients with cirrhosis. Eight subjects were treated with 25 mg of pyridoxine for 28 days. Before and after the supplementation period, B6 status was assessed by measuring fasting plasma vitamer levels and response to a 25 mg oral pyridoxine load. In addition, a 24-hr urine collection was analyzed during each load study for B6 metabolites. The data indicated that supplementation achieved repletion of peripheral B6 stores, as evidenced by: (i) a significant (p < 0.005) rise in fasting plasma pyridoxal phosphate after supplementation (mean ± S.D. = 56.8 ± 30.5 nmoles per liter) as compared to initial levels (17.0 ± 17.8 nmoles per liter); (ii) a higher (p < 0.05) percentage excretion of the pyridoxine load as urinary 4-pyridoxic acid (31.0 ± 9.3%) compared to the initial load (19.6 ± 5.8%), and (iii) a postsupplementation area under the plasma concentration vs. time curve for pyridoxal phosphate (377 ± 529 nmoles·hr per liter), which was decreased (p < 0.005) from the presupplementation value (934 ± 756 nmoles·hr per liter). The postsupplementation fasting plasma pyridoxal phosphate concentrations were within the normal range. The consequences of B6 repletion on amino acid metabolism were measured by oral protein loads (n = 4) or oral methionine loads (n = 4). No significant changes were observed for methionine or any other amino acid in regard to plasma fasting concentration, peak concentration or AUC. Although the vitamin B6 deficiency of cirrhosis was corrected by daily oral pyridoxine supplementation, there was apparently no improvement in the deranged amino acid metabolism.
Hepatic encephalopathy in thioacetamide-induced acute liver failure in rats: Characterization of an improved model and study of amino acid-ergic neurotransmission†
Authors: Zimmermann Christof , Ferenci Peter , Pifl Christian , Yurdaydin Cihan , Ebner Josef , Lassmann Hans , Roth Erich , Hörtnagl Heide
Published: March 2007

An imbalance of excitatory and inhibitory amino acid-ergic neurotransmission has been suggested to play a role in the pathogenesis of hepatic encephalopathy. For further evaluation of this hypothesis, several parameters of amino acid-ergic neurotransmission were studied in rats with acute liver failure induced by the administration of 300 mg per kg thioacetamide by gavage on two consecutive days. By appropriate supportive care, hypoglycemia, renal failure and hypothermia were avoided. Rats were monitored clinically and neurologically. Hepatic encephalopathy evolved in four distinct, easily recognizable stages. Light and electron microscopic examination of brains of rats with hepatic encephalopathy revealed only a slight swelling of nuclei of neurons and astrocytes without signs of neuronal degeneration or brain edema.

In rats with hepatic encephalopathy, the concentrations of GABA, glutamate and taurine were decreased in the cerebral cortex, the hippocampus and the stria-tum, whereas those of aspartate and glycine were unchanged or increased.

GABAA and benzodiazepine receptors were studied as parameters for the postsynaptic GABAA-benzodiazepine receptor complex, glutamic acid decarboxylase as parameter for presynaptic GABA-ergic neurons and stimulation of benzodiazepine binding by GABA as a parameter for a GABA-mediated postsynaptic event. None of these parameters was different in hepatic encephalopathy as compared to controls. Similarly, Ca++/Cl−-dependent and -independent glutamate receptors as parameters for glutamatergic neurons were unchanged in rats with hepatic encephalopathy.

Thus, in rats with thioacetamide-induced liver failure and hepatic encephalopathy, changes of the concentrations of neurotransmitter amino acids occur in the brain. Other neurochemical parameters, however, failed to identify alterations of GABA-ergic or glutamatergic neurotransmission in hepatic encephalopathy.
The aminopyrine breath test predicts the outcome of hepatic transplantation in pigs
Authors: Oakland Christian D. H. , Hickman Rosemary , Terblanche John
Published: March 2007

The aminopyrine breath test has been performed on Day 1 and Day 7 in pigs following experimental liver allografting. The results show that at 24 hr after surgery there was a significant reduction in liver function in all animals as measured by the elimination rate constant of 14CO2 in the breath. This was very profound in the group that died within 5 days (0.09 ± 0.01 hr−1) and was significantly different from the group that survived (0.16 ± 0.01 hr−1). There appears to be a critical value of the elimination rate constant ( = 0.12 hr−1) which predicts death within 5 days. These results would justify a trial of the aminopyrine breath test in patients after liver transplantation.
Impaired liver function in stable renal allograft recipients
Authors: Frey Felix J. , Schaad Heinz J. , Renner Eberhard L. , Horber Fritz F. , Frey Brigitte M. , Preisig Rudolf
Published: March 2007

Hepatic failure as a cause of death is increased in stable renal allograft recipients when compared with patients on dialysis. In order to assess the magnitude and the natural history of the hepatic functional derangement, the kinetics of xenobiotics which are metabolized by cytosolic (galactose) or microsomal (prednisolone, cyclosporine A) enzymes were determined in 28 consecutive stable kidney transplant patients 1 month and 1 year after transplantation. Renal transplant patients had a decreased mean (± S.D.) galactose elimination capacity at 1 month (6.26 ± 0.94 mg per min × kg) and at 1 year (5.93 ± 0.96 mg per min × kg), when compared with a different group of 28 healthy control subjects (7.52 ± 0.78 mg per min × kg, p < 0.001) and a decreased total body clearance of prednisolone at 1 month (2.13 ± 0.34 ml per min × kg vs. 2.71 ± 0.43 ml per min × kg in controls, p < 0.001), which further decreased over the following year to 1.76 ± 0.32 ml per min × kg (p < 0.001). The clearance of cyclosporine A declined significantly during the first year of successful transplantation (5.9 ± 2.1 ml per min × kg vs. 4.9 ± 1.2 ml per min × kg, p < 0.05). In conclusion, a substantial proportion of stable renal transplant recipients have decreased cytosolic and microsomal liver functions despite the absence of clinical and laboratory evidence of significant liver disease.
The effect of estrogen and tamoxifen on hepatocyte proliferation in Vivo and in Vitro
Authors: Francavilla Antonio , Polimeno Lorenzo , Dileo Alfredo , Barone Michele , Ove Peter , Coetzee Mona , Eagon Patricia , Makowka Leonard , Ambrosino Giovanni , Mazzaferro Vincenzo , Starzl Thomas E.
Published: March 2007

We have previously shown that changes in estrogen-hepatocyte interaction occur during liver regeneration. Following 70% hepatectomy, estrogen levels in the blood were elevated, the number of estrogen receptors in the liver was increased and there was an active translocation of estrogen receptors from the cytosol to the nucleus. The injection of tamoxifen, an estrogen antagonist, inhibits hepatocyte proliferation following partial hepatectomy. The administration of 1 μg tamoxifen per gm body weight at zero time or 6 hr after the operation resulted in a significant inhibition both of DNA synthesis and of the number of cells in mitosis. Injections of tamoxifen 12 hr or later after the operation had no effect. Concomitant injections of equimolar amounts of estrogen abolished the inhibition by tamoxifen. The effects of estrogen and tamoxifen were also tested on hepatocytes in primary culture. Estrogens in the presence of 5% normal rat serum stimulated hepatocyte DNA synthesis as determined by [3H]thymidine incorporation and the labeling index, whereas epidermal growth factor-induced DNA synthesis in the absence of normal rat serum was strongly inhibited. Tamoxifen, in contrast, inhibited DNA synthesis of hepatocytes in the presence of 5% normal rat serum and reversed the stimulatory effect of estrogen in the same system. Attempts to elucidate the mechanism of tamoxifen inhibition in vitro indicated that one effect of tamoxifen is to prevent the amiloride-sensitive Na+ influx necessary to initiate hepatocyte proliferation.
Experimental systems for the study of hepadnavirus and hepatitis delta virus infections
Authors: Mason William S. , Taylor John M.
Published: March 2007

The past decade has seen a dramatic increase in the number of approaches available for the study of hepadnavirus and hepatitis delta virus infections. In this review, we have summarized the recent applications of these approaches to the study of virus replication, tissue specificity, liver injury and hepatocellular carcinogenesis.
Colchicine treatment of cirrhosis: Questions
Authors: Grace Norman D.
Published: March 2007

There is preliminary evidence that colchicine, an inhibitor of collagen synthesis, may be beneficial in the treatment of cirrhosis of the liver. To evaluate the use of colchicine (1 mg per day, five days per week) in the treatment of hepatic cirrhosis, we performed a randomized, double-blind, placebo-controlled trial in which 100 patients were followed for up to 14 years. Forty-five patients had alcoholic cirrhosis, 41 had posthepatitic cirrhosis, and the remaining 14 had cirrhosis with various other causes. Histologic studies were available for 92 percent of patients. Seventy-three patients were in Child-Turcotte class A, 26 were in class B, and one was in class C.

Fifty-four patients received colchicine, and 46 received placebo. The overall survival in the colchicine group was markedly better than in the placebo group (median survival, 11 and 3.5 years, respectively; P < 0.001). The cumulative 5-year survival rates were 75 percent in the colchicine group and 34 percent in the placebo group; the corresponding 10-year survival rates were 56 percent and 20 percent. Among the 30 patients treated with colchicine who underwent repeated liver biopsies, histologic improvement was seen in 9; the liver appeared normal in 2, and 7 had minimal portal fibrosis. No histologic improvement was observed in the 14 members of the placebo group who had two or more biopsies. Few side effects were observed in either group.
Hepatic mitochondrial oxidative metabolism and lipid peroxidation in experimental hexachlorobenzene-induced porphyria with dietary carbonyl iron overload†
Authors: Feldman Edward S. , Bacon Bruce R.
Published: March 2007

Both human porphyria cutanea tarda and experimental hexachlorobenzene-induced porphyria are associated with hepatic injury and are potentiated by excess hepatic iron. The mechanisms whereby cellular injury occurs and the synergistic role of iron overload are unknown. In the present experiments, we studied hepatic mitochondrial function and lipid peroxidation in rats with hexachlorobenzene-induced porphyria in which iron loading was achieved by dietary carbonyl iron supplementation. Female rats were treated for 8 weeks, receiving a chow diet supplemented with hexachlorobenzene (0.2%, w/w), carbonyl iron (1.0%, w/w) or hexachlorobenzene + iron. Hepatic total porphyrins were increased 100-fold in rats receiving hexachlorobenzene (hexachlorobenzene alone and hexachlorobenzene + Fe), and total hepatic iron was increased approximately 10-fold in rats receiving iron supplementation (Fe alone and hexachlorobenzene + Fe). There was a significant increase in mitochondrial lipid peroxidation in rats treated with hexachlorobenzene alone and hexachlorobenzene + Fe. A significant reduction in mitochondrial respiratory control ratios and in oxidative phosphorylation (ADP/O ratios) using glutamate and succinate as substrates was demonstrated when rats were treated with hexachlorobenzene + iron. The reductions in respiratory control ratios were due to a combination of an inhibitory defect in electron transport as evidenced by an irreversible decrease in State 3 respiration and an uncoupling effect as evidenced by an increase in State 4 respiration. These findings suggest that lipid peroxidation and mitochondrial dysfunction may contribute to the hepatotoxicity seen in hexachlorobenzene-induced porphyria.
Liver transplantation: Intraoperative changes in coagulation factors in 100 first transplants
Authors: Lewis Jessica H. , Bontempo Franklin A. , Awad Sami A. , Kang Yoo Goo , Kiss Joseph E. , Ragni Margaret V. , Spero Joel A. , Starzl Thomas E.
Published: March 2007

Six intraoperative blood samples were obtained at intervals from each of 100 individuals undergoing their first liver transplants. The patients fell into the following diagnostic categories: postnecrotic cirrhosis 28, primary biliary cirrhosis 20, sclerosing cholangitis 19, miscellaneous diseases 14, carcinoma/neoplasia 12 and fulminant hepatitis 7. Coagulation factor values in the initial (baseline) blood samples varied by patient diagnosis. In general, all factor levels were reduced except factor VIII:C, which was increased to almost twice normal. The slight intraoperative changes in factors II, VII, IX, X, XI and XII suggested that a steady-state relationship existed between depletion (consumption/bleeding) and repletion (transfusion, transit from extra- to intravascular space), even in the anhepatic state. In contrast, there were rapid and very significant falls in factor VIII and fibrinogen and a less pronounced decrease in factor V, all reaching their nadirs in early to mid-Stage III. The cause of these coagulation changes appears to be activation of the fibrinolytic system.
Biologic significance of the detection of HBsAg and HBcAg in liver and tumor from 204 HBsAg-positive patients with primary hepatocellular carcinoma
Authors: Hsu Hey-Chi , Wu Tsung-Teh , Sheu Jin-Chuan , Wu Chieng-Yen , Chiou Tzyy-Jen , Lee Chu-Shue , Chen Ding-Shinn
Published: March 2007

Hepatitis B virus surface and core antigens (HBsAg, HBcAg) were examined in the resected primary hepatocellular carcinoma from 204 patients who had HBsAg in serum. Ninety patients had small (<5 cm) and 114 had large hepatocellular carcinoma (>5 cm). HBsAg was detected in hepatocellular carcinoma in 65 cases (32%) and HBcAg in 30 cases (14.7%); hepatitis B virus antigens were more frequently detected in small (HBsAg in 42.2% and HBcAg in 20%) than in large hepatocellular carcinoma (HBsAg 23.7% and HBcAg 10.5%). These results suggest that replicative forms of hepatitis B virus DNA may exist in hepatocellular carcinoma more frequently than previously believed and that the malignant hepatocytes can support hepatitis B virus replication. A lymphocytic infiltration in hepatocellular carcinoma was more often observed in hepatocellular carcinoma expressing HBsAg (71%) or HBcAg (63%) than in hepatocellular carcinoma with no detectable HBsAg (26%) or HBcAg (37%), p < 0.01. The reaction was mild in the majority (85%) of the cases. These findings suggest that hepatitis B virus antigen expression in hepatocellular carcinoma can provoke a local immune response. The most striking finding was that patients with hepatitis B virus antigens in small hepatocellular carcinoma had a 5-year survival rate (13%) lower than that (50%) of the antigen-negative patients (p < 0.05). In contrast, patients with a marked local immune response in hepatocellular carcinoma, regardless of the viral antigen status, had significantly better 5-year survival rates (43%) than those with no or a mild lymphocytic reaction (18%). These findings indicate that a marked immune response in hepatocellular carcinoma is a favorable prognostic sign. The unfavorable course of the patient with viral antigen-expressing hepatocellular carcinoma may in fact reflect the inability of the host immune response to clear the viral infection and the antigen-expressing tumor cells.
Poly(A+)RNA levels of growth-, differentiation- and transformation-associated genes in the progressive development of hepatocellular carcinoma in the rat
Authors: Huber Brian E. , Heilman Carole A. , Thorgeirsson Snorril S.
Published: March 2007

The development of chemically induced hepatocellular carcinoma in the rat proceeds through a series of premalignant changes that may ultimately progress to a primary malignant tumor. Using the selection technique based on diminished binding of preneoplasic hepatocytes to tissue culture plates precoated with asialofetuin, we have isolated poly(A+)RNA from early preneoplastic foci as well as preneoplastic persistent nodules and primary hepatocellular carcinoma induced by the Solt-Farber protocol in the Fischer rat. The steady-state poly(A+)RNA levels of genes traditionally associated with growth, differentiation and/or transformation were then determined to address the question of their temporal expression in the multistep nature of cancer development.

Ornithine decarboxylase- and P53-specific transcripts did not significantly change in preneoplastic foci but were increased in later-stage preneoplastic nodules and hepatocellular carcinoma. Albumin-specific transcripts were decreased in all hepatocellular carcinoma but there was no consistent coordinated increase in α-fetoprotein-specific transcripts. c-myc and raf transcripts increased at the very early preneoplastic foci stage and continued to increase throughout the neoplastic process. No L-myc or N-myc transcripts could be detected in any RNA sample. c-Ha-ras-specific transcripts were essentially unaltered in all RNA samples whereas no c-Ki-ras or N-ras transcripts could be detected throughout the neoplastic process. In addition, no dominant-acting transforming mutations in the ras gene family were detected by DNA transfection experiments using NIH/3T3 cells.
What is the place of laser lithotripsy in the treatment of common bile duct stones?
Authors: Saurebruch T.
Published: March 2007

Endoscopic retrograde laser lithotripsy of common bile duct stones is a new technique which can be carried out through the endoscope without anaesthesia using ordinary endoscopic equipment. In the method described here a flashlamp pulsed Neodymium YAG laser (wave length 1064mm) was used. Light energy was transmitted along a highly flexible quartz fibre with a diameter of 0.2mm. This new technique was used in nine patients with concrements in the common bile duct, which could not be removed with the established endoscopic techniques. In eight of the nine the concrements (maximum diameter 4.7 × 3.1 cm) could be fragmented and in six the fragments could be extracted from the common bile duct. The total energy required was 80-300J; complications were not observed.
Endotoxin-induced hypercoagulability: A possible aggravating factor of alcoholic liver disease†
Authors: Arai Masao , Nakano Shigeo , Okuno Fumio , Hirano Yoshiaki , Sujita Kazufumi , Kobayashi Toshiji , Ishii Hiromasa , Tsuchiya Masaharu
Published: March 2007

The present experiments were designed to study the effect of chronic ethanol consumption on endotoxin toxicity. The intravenous injection of endotoxin produced a more pronounced increase of serum AST and ALT activities in chronic ethanol-fed rats, when compared to controls. The activities of hepatic mitochondrial enzymes, succinate dehydrogenase and cytochrome oxidase, were also distinctly decreased by endotoxin treatment in chronic ethanol-fed rats. Consistent with these biochemical alterations, light and electron microscopic examinations revealed severe liver injury after endotoxin injection in chronic ethanol-fed rats. Furthermore, the increase of blood BUN and creatinine levels accompanied by the degeneration of the renal tubulus and slight infiltration of neutrophils into the glomerule were produced by endotoxin treatment and were more conspicuous in chronic ethanol-fed rats than controls. Therefore, the biochemical and histological evidence indicates that endotoxin markedly potentiates organ injury after chronic ethanol consumption. In addition, a more pronounced decrease in blood antithrombin III activity accompanied by an increase in fibrin degradation product level in blood was recognized in chronic ethanol-fed rats receiving endotoxin, when compared to controls receiving endotoxin. This increase of blood fibrin degradation product level correlated well with the decrease of antithrombin III activity (r = −0.6116; p < 0.005). These findings of blood antithrombin III activity and fibrin degradation product level indicate that the coagulation-fibrinolysis system is more activated by endotoxin treatment after chronic ethanol consumption. Furthermore, the activation of the coagulation-fibrinolysis system was well correlated with biochemical and histological alterations representing hepatorenal involvement. Therefore, the results from the present study indicate that chronic ethanol consumption potentiates endotoxin-induced organ injury and that hypersensitive activity to endotoxin of the coagulation-fibrinolysis system after chronic ethanol consumption may, at least in part, pathogenically contribute to the aggravation of organ injury.
Selective hepatobiliary transport of nordeoxycholate side chain conjugates in mutant rats with a canalicular transport defect
Authors: Elferink Ronald P. J. Oude , de Haan Jan , Lambert Karel J. , Hagey Lee R. , Hofmann Alan F. , Jansen Peter L. M.
Published: March 2007

Canalicular transport of bilirubin diglucuronide, dibromosulfophthalein and several glutathione conjugates is deficient in mutant TR− rats. In contrast, transport of cholyltaurine (taurocholate), a conjugated bile acid, is normal. Previous studies using normal rats have shown that C23 nor-dihydroxy bile acids are conjugated with sulfate or glucuronide during hepatic transport in contrast to the natural C24 bile acids, which are amidated with glycine or taurine. Studies were performed to test the hypothesis that (a) in the TR− rat, nordeoxycholate would be conjugated with glucuronate or sulfate just as in the normal rat, and (b) that such conjugates would have defective biliary secretion. [C23-14C]Nordeoxycholate was administered intravenously to bile fistula rats (TR− and normal), and the biliary recovery of metabolites was assessed by chromatography and mass spectrometry.

In both groups of rats, the major biotransformation product of nordeoxycholate was the side chain (23-ester) glucuronide. Conjugation on the nucleus with sulfate and glucuronide at the 3-position (ethereal linkage) also occurred, as well as amidation at the C23 carboxylic acid group. In the mutant rats, biliary secretion of the 3-sulfate and 3-glucuronide conjugates was less than 10% and 1%, respectively, of that of normal rats, whereas biliary secretion of the 23-ester glucuronide and the 23-taurine amidate, as well as unchanged nordeoxycholate, was not decreased. Canalicular secretion of nor-bile acid 3-ether glucuronides and 3-sulfates appears to involve the “bilirubin transport system,” which is deficient in mutant rats. Canalicular secretion of unconjugated, amidated or esterified nordeoxycholate is mediated via another pathway, probably the “bile acid transport system.” It is speculated that the “bile acid transport system” transports large organic anions with one (or more) negative charges on one side chain (“unipolar anions”), whereas the “bilirubin transport system” transports large organic anions with at least two separate negative side chains tentatively termed “multipolar anions,” which include the 3-sulfates and 3-glucuronides of bile acids.
Delta hepatitis in homosexual men in the united states
Authors: Weisfuse Isaac B. , Hadler Stephen C. , Fields Howard A. , Alter Miriam J. , O’Malley Paul M. , Judson Franklyn N. , Ostrow David G. , Altman Norman L.
Published: March 2007

To assess the incidence and prevalence of delta hepatitis in homosexual men, we tested serum specimens for delta markers in participants in two previous studies: a hepatitis B vaccine trial among homosexual men conducted in the early 1980′s and the Centers for Disease Control sentinel counties hepatitis study for 1983-1984. In the vaccine trial, men found to be hepatitis B surface antigen positive at the time of enrollment and those men who had serologic evidence of new hepatitis B virus infection during follow-up were tested. In the sentinel counties study that determined risk factors for viral hepatitis in reported cases, all homosexual men with acute and chronic hepatitis B virus infections were tested for delta markers. Specimens were tested for delta antigen and IgM and total delta antibody. In seven different cities, among 321 men found to be HBsAg positive at the time of screening, eight (2%) were positive for any delta marker. Among 290 men with new hepatitis B virus infections during follow-up, three (two coinfections, one superinfection) had serologic evidence of delta hepatitis. In the sentinel counties study, 0/63 acute hepatitis B virus infections in homosexual men were associated with delta hepatitis. This study indicates that the delta agent is an infrequent cause of viral hepatitis in homosexual men in the United States.
Fat-storing cells and myofibroblasts: One cell or two?
Authors: Maher Jacquelyn J.
Published: March 2007

To differentiate cultured rat liver myofibroblasts, fat-storing cells, aortic smooth muscle cells and skin fibroblasts from each other, desmin and vimentin stainings were undertaken by indirect immunofluorescence using monoclonal antibodies. In myofibroblasts, the reaction with antibodies to vimentin was positive but that with antibodies to desmin was virtually negative. In primary cultures as well as subsequent passage of fat-storing cells, reactions with antibodies to both desmin and vimentin were positive. In primary culture of smooth muscle cells, both reactions were positive, but in the first passage, smooth muscle cells lost the reactivity with antibodies to desmin. Fibroblasts showed a positive reaction with antibodies to vimentin and a negative one with antibodies to desmin. Thus, immunohistochemistry of intermediate filaments allows for the differentiation between fat-storing cells, which are desmin- and vimentin-positive, and myofibroblasts or fibroblasts, which are desmin-negative but vimentin-positive. Smooth muscle cells are also vimentin-positive and become desmin-negative after the first passage.
The initial hemorrhage from esophageal varices
Authors: Conn Harold O.
Published: March 2007

We conducted a prospective study of 321 patients with cirrhosis of the liver and esophageal varices with no history of bleeding to see whether a comprehensive analysis of their clinical features and of the endoscopic appearances of their varices could help to identify those at highest risk for bleeding. Varices were classified endoscopically as suggested by the Japanese Research Society for Portal Hypertension. Patients were followed for 1 to 38 months (median, 23), during which 85 patients (26.5 percent) bled. Multiple regression analysis (Cox’s model) revealed that the risk of bleeding was significantly related to the patient’s modified Child class (an index of liver dysfunction based on serum albumin concentration, bilirubin level, prothrombin time, and the presence of ascites and encephalopathy), the size of the varices, and the presence of red wale markings (longitudinal dilated venules resembling whip marks) on the varices. A prognostic index based on these variables was devised that enabled us to identify a subset of patients with a one-year incidence of bleeding exceeding 65 percent. The index was prospective validated on an independent sample of 75 patients with varices and no history of bleeding. We conclude that our prognostic index, which identifies groups of patients with one-year probabilities of bleeding ranging from 6 to 76 percent, can be used to identify candidates for prophylactic treatment.