bluuebunny/crossref_metadata_2025 huggingface dataset:
The fasting B6 vitamer profile and response to a pyridoxine load in normal and cirrhotic subjects
Authors: Henderson Michael J. , Codner Mark A. , Hollins Bettye , Kutner Michael H. , Merrill Alfred H.
Published: March 2007

This study established the fasting plasma and urine profiles of vitamin B6 in cirrhotics and assessed the response to an oral dose of pyridoxine. High-performance liquid chromatography was used to measure all vitameric coenzymatic and degradatory forms. In 31 patients with cirrhosis and 15 healthy controls, fasting plasma and 24-hr urine collection showed: plasma pyridoxal-5′-phosphate, the biologically active form, was significantly (p<0.001) reduced in cirrhotics (mean ± S.D.: 5.7 ± 3.2 ng per ml) compared to normals (14.2 ± 7.5 ng per ml); plasma pyridoxal was detected in more cirrhotics (48%) than normals (28%); pyridoxic acid, the end catabolite, was significantly (p<0.05) lower in plasma of cirrhotics compared to normals, but 24-hr urine excretion was not different. Administration of 25 mg of pyridoxine to 7 cirrhotics and 5 normals showed the following plasma changes: (i) pyridoxine rapidly peaked at 30 min and was totally cleared from plasma by 3 hr; (ii) plasma pyridoxal and pyridoxic acid increased in parallel up to 40-fold over baseline by 1 to 2 hr and rapidly fell toward baseline by 8 hr, and (iii) plasma pyridoxal-5′-phosphate, in contrast, increased significantly (p<0.05) from baseline by 60 min and was maintained above normal for 24 hr. The area under the plasma concentration vs. time curve (AUC) for pyridoxal-5′-phosphate was significantly (p<0.05) less for the cirrhotics than normals and showed a significant negative correlation to hepatocyte function and blood flow. This suggests that it is most difficult to replete plasma pyridoxal-5′-phosphate in those cirrhotics with best preservation of liver function and flow. In contrast, the plasma pyridoxal AUC is significantly (p<0.025) greater in the cirrhotics after the pyridoxine load. Plasma pyridoxic acid AUC and urine pyridoxic acid excretion (27% of load per 24 hr) are not significantly different for the two groups. The data are consistent with normal pyridoxal-5′-phosphate synthesis since plasma pyridoxal-5′-phosphate levels are maintained within the normal range by a single 25 mg oral dose of pyridoxine. However, the smaller AUC for pyridoxal-5′-phosphate in cirrhotics compared to normals, combined with the increased pyridoxal AUC, shows increased bydrolysis to pyridoxal. This is not excreted as pyridoxic acid and suggests that pyridoxal may be entering intracellular sites.
Genetic polymorphism of human liver alcohol and aldehyde dehydrogenases, and their relationship to alcohol metabolism and alcoholism
Authors: Bosron William F. , Li Ting-Kai
Published: March 2007

It is now widely accepted that the various pharmacologic and addictive consequences of alcohol consumption are related to the tissue concentration of ethanol or its metabolic products. The oxidative metabolism of ethanol in liver is principally catalyzed by alcohol dehydrogenase and aldehyde dehydrogenase. Both of these enzymes exist in multiple molecular forms, and genetic models have been proposed to account for the multiplicity of isoenzymes. Alcohol dehydrogenase subunits are encoded at five different gene loci, and genetic polymorphism occurs at two alcohol dehydrogenase loci. Variant isoenzymes produced at the two polymorphic alcohol dehydrogenase loci account for the differences in enzyme electrophoretic patterns observed among individuals. Some of these variant isoenzymes exhibit widely different kinetic properties, and this may account for the 2- to 3-fold variation in alcohol elimination rate among individuals. Since the protein sequence of several of the alcohol dehydrogenase subunits has been determined and several of the alcohol dehydrogenase genes has been cloned, some of the structural changes which give rise to differences in catalytic and electrophoretic properties are now known. Genetic polymorphism also occurs at the aldehyde dehydrogenase gene locus which encodes the mitochondrial low Km for acetaldehyde aldehyde dehydrogenase isoenzyme. The variant isoenzyme exhibits little or no catalytic activity. Individuals with this “null” variant have higher than normal blood acetaldehyde levels and exhibit an alcohol-flush reaction which appears to be a deterrent to heavy drinking and alcoholism. Since epidemiological surveys have demonstrated genetic predisposition to alcohol abuse and alcoholism, it is reasonable to suppose that the polymorphism of either alcohol dehydrogenase or aldehyde dehydrogenase might be one of the underlying biological factors contributing to the inherited nature of alcoholism.
Transgenic mouse models for the chronic hepatitis B surface antigen carrier state: Transfection a deux
Authors: Di Bisceglie Adrian M. , Hoofnagle Jay H.
Published: March 2007

In an attempt to establish a model of the healthy carrier state in hepatitis B virus (HBV) infections, transgenic mice expressing HBV genes were produced. Fertilized one-cell eggs were microinjected with subgenomic fragments of HBV DNA containing the coding regions for the HBV surface antigen (HBsAg) and pre-S and×antigens. Either the normal (HBV) or metallothionein promoters were used to obtain expression of the HBV genes. There was no evidence of viral replication or tissue pathology. The integrated HBV DNA sequences were inherited in a normal Mendelian fashion. Three of 16 transgenic mice expressed HBV-encoded gene products to which they were immunologically tolerant. Expression was not tissue specific and may be influenced by the genomic integration site and cellular factors. Both HBsAg and pre-S antigen were detectable within the cytoplasm of hepatocytes and renal tubular epithelial cells. High serum concentrations of HBsAg were detectable and the secreted product appeared authentic as judged by mean density, morphology, mean particle diameter, polypeptide composition, and antigenicity. The absence of tissue pathology in these immunologically tolerant animals supports the hypothesis that cellular injury under these conditions is not a direct consequence of expression of the pre-S or HBs regions of the HBV genome.

Two transgenic mice were obtained that contain in their chromosomes the complete hepatitis B virus (HBV) genome except for the core gene. These mice secrete particles of HBV surface antigen (HBsAg) in the serum. In one mouse, HBV DNA sequences that had integrated at two different sites were shown to segregate independently in the first filial generation (F1) and only one of the sequences allowed expression of the surface antigen. Among these animals the males produced five to ten times more HBsAg than the females. A 2.1-kilobase messenger RNA species comigrating with the major surface gene messenger RNA is expressed specifically in the liver in the two original mice. The results suggest that the HBV sequences introduced into the mice are able to confer a tissue-specific expression to the S gene. In addition, the HBV transgenic mice represent a new model for the chronic carrier state of hepatitis B virus infection.
Incarcerate free radicals
Authors: Granger Neil D. , Parks Dale A.
Published: March 2007

We have investigated the possible protective effect of superoxide dismutase and allopurinol in a rat model of mild and severe hepatic necrosis produced by Corynebacterium parvum with or without endotoxin. Histology showed a sinusoidal mononuclear cell infiltrate with multiple granulomata but variable degrees of hepatic necrosis. In the severe hepatic injury model there was a reduction in mortality, associated with a decrease in histologic and biochemical evidence of hepatic necrosis, after treatment with superoxide dismutase. This protective effect was not demonstrated with partially heat-inactivated superoxide dismutase. In the mild hepatic injury model similar trends in reduction of serum levels of hepatic enzymes were observed after treatment with both superoxide dismutase and allopurinol. These results indicate that oxygen-derived free radicals may play an important role in the pathogenesis of hepatic injury in the rat.
Characterization and clinical relevance of a new complement-fixing antibod-anti-m8-in patients with primary biliary cirrhosis
Authors: Weber Paul , Brenner Judith , Stechemesser Ernst , Klein Reinhild , Weckenmann Ulrike , Klöppel Günter , Kirchhof Manfred , Fintelmann Volker , Berg Peter A.
Published: March 2007

A new complement-fixing antimitochondrial antibody-anti-M8-was detected in patients with primary biliary cirrhosis. Anti-M8 was only found in association with anti-M2, however, not all anti-M2 positive patients had anti-M8. Thus, among 66 anti-M2 positive patients, 29 were also positive for anti-M8, whereas sera from patients who had the complement-fixing anti-M2 and anti-M4 antibodies in parallel always strongly reacted with the M8 antigen. This group was previously described as mixed form. The M8 antigen was isolated either from human liver mitochondria or pig kidney microsomes and could be clearly distinguished from the M4 antigen. In contrast to M4, M8 was trypsin sensitive and banded at sucrose densities from 1.16 to 1.24, while M4 was found at densities from 1.08 to 1.14. Like M4, the M8 antigen also co-purified with outer mitochondrial membranes.

Fifty-three patients with primary biliary cirrhosis have been followed over a period of up to 16 years and were classified according to their complement-fixing antimitochondrial antibody profile. At the time of the first diagnosis, 95% of 31 patients being anti-M2 positive, but anti-M8 negative (antimitochondrial antibody Profile I) were in Stage I or II. In contrast, only 61% of 13 patients being anti-M2 and anti-M8 positive (antimitochondrial antibody Profile II) and 44% of 9 patients with anti-M2, anti-M8 and anti-M4 in parallel (antimitochondrial antibody Profile III) belonged to Stage I or II. At the end of the observation period, only 24% of patients with antimitochondrial antibody Profile I (anti-M8 negative) had reached Stage III or IV, while 75% of patients with antimitochondrial antibody Profile II or III (anti-M8 positive) had a progression to Stage III or IV.

It is concluded that the presence of complement-fixing anti-M8 antibody may herald a more progressive course of primary biliary cirrhosis.
Estrogen and progesterone receptors in human gallbladder
Authors: Singletary Brian K. , van Thiel David H. , Eagon Patricia K.
Published: March 2007

Gallbladder disease is more prevalent in women than men. Estrogen therapy has been associated with an increased incidence of gallbladder disease in both sexes. Further, increased progesterone levels have been implicated in impairment of gallbladder motility in pregnancy. Because sex hormones often exert their action through specific receptors, we investigated whether human gallbladder contains receptors for estrogen and progesterone. Binding of radiolabeled hormones in cytosol and nuclei prepared from human gallbladder of both sexes is indicative of the presence of receptors for both estrogen and progesterone. The binding is saturable, of high affinity and highly specific for its particular type of hormone but not other steroids. Fractionation of sodium molybdate-stabilized gallbladder cytosol on Sephadex G-100 demonstrates that the labeled hormones are not bound to defined proteins such as sex steroid binding globulin or albumin. These studies indicate that human gallbladder contains both estrogen and progesterone receptors, that the presence of these receptors may explain the sensitivity of gallbladder tissue to these hormones and that certain aspects of gallbladder function may be mediated by the interaction of steroid hormones with these receptors.
Incidental solitary hepatocellular carcinomas smaller than 1 cm in size found at autopsy: A morphologic study
Authors: Nakanuma Yasuni , Ohta Goroku , Sugiura Hitoshi , Watanabe Kishichiro , Doishita Kenji
Published: March 2007

The morphologic features and growth pattern of single hepatocellular carcinomas ≤ 1 cm in size, found incidentally at autopsy, were studied in nine cases. In all but one case, the hepatic parenchyma showed advanced cirrhosis. In three cases, the hepatocellular carcinomas were localized within a regenerative nodule as a form of “nodule within nodule.” The carcinoma was rimmed by nonneoplastic hepatic tissue. A fourth carcinoma consisted of an expansile hepatocellular carcinoma nodule enclosed by a fibrous band of cirrhotic stroma. The remaining five cases consisted of hepatocellular carcinomas which infiltrated the surrounding regenerative nodules or hepatic lobules. These observations suggested that hepatocellular carcinomas arise within regenerative nodules, some of which still retain residual nonneoplastic tissue around the tumor. Others invaded the adjacent liver tissue. The grossly visible fibrous capsule, often seen in more advanced hepatocellular carcinomas, was absent in all cases. All of the hepatocellular carcinomas were well-differentiated. Four cases showed a trabecular pattern with slight sinusoidal dilatation, 3 showed a scirrhous pattern and 2 showed a compact pattern. Their histologic features included marked bile production, Mallory body formation by clusters of tumor cells, resistance to hemosiderin deposition in a markedly siderotic background and loss or decrease of reticulin fibers. These features were hallmarks of small hepatocellular carcinomas. Pathologists should study cirrhotic livers carefully so as not to miss small carcinomas. Clinicians should be aware that even small liver nodules may be hepatocellular carcinomas.
Absence of detectable serum interferon in acute and chronic viral hepatitis
Authors: Pirovino Mauro , Aguet Michel , Huber Marcel , Altorfer Josef , Schmid Martin
Published: March 2007

Amounts of interferon were measured in sera from 59 patients with acute viral hepatitis and 49 patients with chronic hepatitis B and compared to those from patients with nonviral liver disease or influenza, and from healthy controls. In all patients with acute and chronic viral hepatitis, no serum interferon could be detected, confirming data from earlier studies of acute viral hepatitis in which no circulating interferon was found. Our results disprove the view that the amounts of serum interferon, detected at the time of the acute clinical illness, may be a determinant of outcome.
Evidence for the role of ammonia in the intracerebral transfer and metabolism of tryptophan
Authors: Grippon Patrick , le Poncin Lafitte Monique , Boschat Michèle , Wang Shwen , Faure Grazyna , Dutertre Dominique , Opolon Pierre
Published: March 2007

After portacaval shunt in the rat, the transport of tryptophan and other neutral amino acids across the blood-brain barrier is enhanced. To determine the role of NH3 in the intracerebral transfer of tryptophan and serotonin metabolism, solutions containing either saline or NH3 or tryptophan or NH3 + tryptophan together were infused, respectively, into the internal carotid artery of rats in order to achieve blood levels similar to those observed after liver ischemia. After tryptophan infusion, a significant increase in the hypothalamic levels of tryptophan and 5-hydroxyindoleacetic acid was observed. A similar increment was found after NH3 infusion. NH3 + tryptophan infusion induced a significant increment in hypothalamic tryptophan and 5-hydroxyindoleacetic acid levels which were 2-fold greater than after tryptophan infusion. There was no significant change in 5-hydroxytryptamine levels in any of these experiments. Glutamine levels increased significantly after NH3 infusion. When tryptophan and NH3 were infused simultaneously, a significant reduction in glutamine levels occurred. These results cannot be explained by any modification of cerebral blood flow nor of the cerebral intratissular pH. Our data suggest that NH3 enhances the transfer of tryptophan across the blood-brain barrier and thus stimulates serotonin metabolism. The mechanism by which tryptophan transfer across the blood-brain barrier is facilated is unknown. The reduction in glutamine levels in the hypothalamus when NH3 and tryptophan are infused together may be explained either by an inhibition of synthesis or by an intratissular influx of neutral amino acids and an efflux of glutamine as suggested by James et al.
Nodular regenerative hyperplasia of the liver associated with the toxic oil syndrome: Report of five cases
Authors: Solis-Herruzo José A. , Vidal José V. , Colina Francisco , Santalla Fermin , Castellano Gregorio
Published: March 2007

Nodular regenerative hyperplasia of the liver developed in five patients who had the toxic oil syndrome caused by ingestion of adulterated cooking oil. This hepatic complication was detected from 19 to 37 months (mean-2.5 years) after the onset of the toxic oil syndrome. Nodular regenerative hyperplasia was asymptomatic, although all patients had persistently abnormal liver function. Hepatomegaly was present in four cases, mild jaundice in three and signs of portal hypertension in two. Pathogenesis of nodular regenerative hyperplasia in toxic oil syndrome is unknown, but probably microcirculatory disturbances within the liver might have played a role. Fibrosis in Zone 3, sinusoidal dilatation and occasionally intralobular hemorrhage were seen in three cases; in one of them, characteristic lesions of venoocclusive disease were also present. In another case, endarteritic changes of hepatic arterioles were evident.
The fluidity of liver plasma membranes from patients with different types of liver injury
Authors: Schuller Amador , Solis-Herruzo José A. , Moscat Jorge , Fernandez-Checa J. C. , Municio Angel M.
Published: March 2007

The physical properties of cell membranes play an important role in cellular homeostasis. Previous studies with animal models have shown that the hepatocyte membrane fluidity has important effects in cellular function. In order to further investigate the relationship between physical properties of membranes and liver disease, the present study was performed. Twenty-nine patients with different degrees of liver injury and 10 control individuals were studied. A lower membrane fluidity-as assessed by diphenylhexatriene fluorescence polarization-was observed in membrane preparations from patients, as compared to controls. These alterations are positively correlated with the severity of liver damage and could be accounted for by a higher cholesterol content of patients' membrane preparations.
Hepatitis B virus is a retrovirus in disguise
Authors: Zeldis Jerome B.
Published: March 2007

Nucleocapsid-pol fusion proteins have been detected by serological screening hepatocellular carcinoma tissues that contain hepatitis B virus (HBV) DNA. The existence of these fusion proteins suggests that HBV may synthesize its reverse transcriptase in a fashion analogous to the way that retroviruses synthesize and process a precursor. The accumulation of HBV reverse transcriptase intermediates in tumorous tissues and not in other tissues may be related to the absence of viral core particles and possibly contributes to tumor development.
Paracetamol hepatotoxicity: A built-in antidote?
Authors: Williams Roger
Published: March 2007

The analgesic effect, toxicity and kinetics of oral paracetamol were compared with those of paracetamol + l-methionine (5:1). The analgesic effect of paracetamol, studied in the Randall-Selitto test in rats, was not changed by methionine: ED50 was 94.6 mg/kg without methionine and 94.1 mg/kg with it. However, methionine reduced the acute toxicity (LD50) of paracetamol by 50% (p < 0.05) in non-fasted, fasted and phenobarbital-pre-treated mice. In a randomized cross-over study in 10 human volunteers the pharmacokinetics of paracetamol (1,500 mg) was not affected by methionine (300 mg). The absorption of methionine from the combination tablets was rapid, peak concentrations occurred in plasma at 30 min and were 3-4 times higher than after paracetamol tablets not containing methionine. Methionine in products containing paracetamol should increase their safety and be the simplest way to reduce the high mortality in paracetamol overdosage.
Glutathione S -transferases: Of rats and men
Authors: Kirsch R. E. , Bass N. M.
Published: March 2007

Rat liver glutathione S-transferases have been purified to apparent electrophoretic homogeneity by S-hexylglutathione-linked Sepharose 6B affinity chromatography and CM-cellulose column chromatography. At least 11 transferase activity peaks can be resolved including five Yb size homodimeric isozymes, two Yc size homodimeric isozymes, one Ya homodimeric isozyme, one Yα homodimeric isozyme, and two Ya-Yc heterodimeric isozymes. Distribution of the GSH peroxidase activity among the CM-cellulose column fractions suggests the existence of further multiplicity in this isozyme family. Substrate specificity patterns of the Yb subunit isozymes revealed a possibility that each of the five Yb-containing isozymes is composed of a different homodimeric Yb size subunit composition. Our findings on the increasing multiplicity of glutathione S-transferase isozymes are consistent with the notion that multiple isozymes of overlapping substrate specificities are required to detoxify a multitude of xenobiotics in addition to serving other important physiological functions.
Bath is a bath is a bath
Authors: Conn Harold O.
Published: March 2007

Immersion in water in spas has been practised for centuries and has many proponents. Despite fierce debate about its efficacy there has been little scientific evaluation of the effect of immersion in mineral waters. Eight normal subjects were immersed in Bath spa water for two hours and the renal, haematological, and cardiovascular responses were compared with those in the control periods before and after immersion. Significant, twofold diuresis and natriuresis, 5% haemodilution, and a 50% increase in cardiac index were observed in subjects immersed, sitting, in Bath spa water at 35°C. These changes may constitute part of the scientific rationale for spa treatment in many states of disease.
Testosterone treatment of men with alcoholic cirrhosis: A double-blind study
Authors: Gluud Christian
Published: March 2007

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.
Ethanol-induced liver fibrosis in rats fed high fat diet
Authors: Tsukamoto Hidekazu , Towner Sally J. , Clofalo Lefran M. , French Samuel W.
Published: March 2007

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.
The combined effects of protein deficiency and chronic ethanol administration on rat ethanol metabolism
Authors: Wilson Jeremy S. , Korsten Mark A. , Lieber Charles S.
Published: March 2007

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.
Receptor-mediated uptake of asialoglycoprotein by the primate liver initiates both lysosomal and transcellular pathways
Authors: Schiff Michael J. , Huling Sandra L. , Jones Albert L.
Published: March 2007

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.
Determination of iron to phosphorus ratios of iron storage compounds in patients with iron overload: A chemical and electron probe X-ray microanalysis
Authors: Cleton Maud I. , Frenkel Erik J. , de Bruijn Willem C. , Marx Joannes J. M.
Published: March 2007

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.
Peripheral T-cell lymphoma presenting as predominant liver disease: A report of three cases
Authors: Gaulard Philippe , Zafrani Elie Serge , Mavier Philippe , Rocha Francisco Dario , Farcet Jean-Pierre , Divine Marine , Haioun Corinne , Pinaudeau Yvon
Published: March 2007

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.
Morphological findings in the liver of children with cystic fibrosis: A light and electron microscopical study
Authors: Hultcrantz Rolf , Mengarelli Silwa , Strandvik Birgitta
Published: March 2007

Liver tissue from five children with cystic fibrosis, obtained through percutaneous liver biopsies, have been investigated via light and electron microscopy. None of the patients had clinical evidence of liver disorder, and their blood chemistry was mainly normal.

Light microscopy showed slight fibrosis in three cases, more advanced fibrosis in one case and focal cirrhotic changes in one case. All patients had fatty infiltration in the hepatocytes and glycogen in the nuclei of these cells.

Electron microscopy showed an increase in the number of Ito cells around the portal tracts and also fibrosis in all patients. In the majority of hepatocytes, no evident necrosis was seen. Hypertrophy of the smooth endoplasmic reticulum and the Golgi apparatus were noted. Large lysosomes containing lipofuscin and lipids were also present. No direct evidence of cholestasis could be seen in the hepatocytes. The bile canaliculi were not dilated and did not contain bile plugs. No bile pigment was seen in the cells, and direct evidence of cholestasis was thus not found in the hepatocytes. Other organelles, such as the rough endoplasmic reticulum, peroxisomes and mitochondria, had a normal appearance. Bile ducts, even when seen in fibrotic portal tracts, were not dilated.

The ultrastructural findings cannot explain the basis for the liver cell damage. Cholestasis does not seem to be a presumable etiological factor as judged from the findings in the present study.
The isolation of functionally heterogeneous hepatocytes of the proximal and distal half of the liver acinus in the rat
Authors: Gumucio Jorge J. , May Maryann , Dvorak Caroline , Chianale Jose , Massey Veronica
Published: March 2007

The objective of this study was to isolate hepatocytes of the proximal half (Zones 1 and 2) or distal half (Zones 2 and 3) of the liver acinus. The zonal origin of the isolated hepatocytes was recognized by: (a) the presence in hepatocytes of a fluorescent marker, acridine orange, selectively delivered to either the proximal or the distal half of the acinus by in situ perfusion prior to cell isolation and (b) the measurement of the induction of cytochrome P-450 by phenobarbital, an induction known to occur predominantly in the distal half of the acinus.

Following the selective labeling of the acinus with acridine orange, livers were perfused with collagenase in either the portal to hepatic vein direction (anterograde) or in the retrograde direction. Hepatocytes isolated by either an anterograde or a retrograde perfusion were separated by centrifugation in a Percoll density gradient. This procedure isolated populations of proximal or distal hepatocytes, respectively, which were intact and 90% fluorescent. In an effort of assessing the heterogeneity of the separated proximal and distal hepatocytes, each population was further fractionated by centrifugal elutriation. This resulted in the arbitrary separation of proximal or distal hepatocytes into five fractions. Total cytochrome P-450 was determined spectrophotometrically in each of the fractions isolated from controls and after 3 days of the in vivo administration of phenobarbital. On the basis of the pattern of fluorescence in isolated hepatocytes and on the cytochrome P-450 inductive response to phenobarbital administration, it is proposed that: (a) the anterograde or retrograde perfusion of the liver with collagenase separated hepatocytes predominantly of the proximal or distal half of the liver acinus, respectively and (b) that hepatocytes of the distal half of the liver acinus responded to phenobarbital administration with the highest level of cytochrome P-450 induction, indicating that the isolated hepatocytes conserved the functional heterogeneity observed in vivo.
Effects of anticonvulsant agents on halothane-induced liver injury in human subjects and experimental animals
Authors: Nomura Fumio , Hatano Hitoshi , Ohnishi Kunihiko , Akikusa Bunshiro , Okuda Kunio
Published: March 2007

In order to evaluate the clinical implication of experimental studies on halothane-induced liver damage in phenobarbital-treated rats, we studied the clinical records of 315 consecutive patients who underwent brain surgery with halothane anesthesia. After exclusion of subjects with a history of alcoholism or antecedent chronic liver disease, clinical data of 279 patients with normal preoperative transaminase activities were analyzed. The incidence of halothane-induced liver injury was significantly higher in the subjects given phenobarbital than in those with no phenobarbital medication (7/100 vs. 1/179, p < 0.01). To determine if other anticonvulsant compounds can influence halothane-induced liver injury, rats were pretreated with diphenylhydantoin or valproic acid prior to exposure to halothane under hypoxic conditions for comparison with phenobarbital. The degree of halothane hepatotoxicity assessed from ALT activities and morphological alterations was of the decreasing order of phenobarbital > controls = diphenylhydantoin > valproic acid, and a similar order was observed in the extent of reductive metabolism of halothane. These results indicate that patients pretreated with phenobarbital may be at a greater risk of halothane-induced liver damage, and that treatment with valproic acid and diphenylhydantoin lead to the production of toxic intermediates of halothane to a lesser extent than treatment with phenobarbital does.
Frequency and significance of antibody to double-stranded DNA in chronic active hepatitis
Authors: Wood James R. , Czaja Albert J. , Beaver Sandra J. , Hall Stephen , Ginsburg William W. , Kaufman David K. , Markowitz Harold
Published: March 2007

To assess the frequency and significance of immunoglobulin G antibody to double-stranded DNA in chronic active hepatitis, 99 patients with severe disease were tested for the antibody by an enzyme-linked immunosorbent assay of established sensitivity and specificity. Antibody was detected in 56 patients (57%) and occurred with similar frequency in patients with autoimmune (64%), idiopathic (46%) and type B (43%) disease. The mean serum level of antibody was higher in autoimmune disease but it was not significantly different from that in the other diagnostic categories. Patients with and without the antibody could not be distinguished by clinical, laboratory or histologic findings. Responses to corticosteroid therapy and mortality were unrelated to antibody status. In five patients, the antibody disappeared after corticosteroid treatment and induction of histologic remission. We conclude that immunoglobulin G antibody to double-stranded DNA is frequently present in patients with severe chronic active hepatitis. Patients with autoimmune disease may have higher antibody levels than others, but the presence of antibody does not connote a disease of specific etiology, unique presentation or different behavior. The prevalence of the finding and its disappearance after corticosteroid therapy suggest that it is a nonspecific manifestation of inflammatory activity.
The in vitro production of antibodies to mitochondrial antigens by peripheral blood mononuclear cells from patients with primary biliary cirrhosis
Authors: Avigan Mark I. , Adamson Gregory , Hoofnagle Jay H. , Jones Anthony E.
Published: March 2007

Peripheral blood mononuclear cells from 7 patients with primary biliary cirrhosis and 7 healthy control subjects were studied for their ability to produce antibodies to mitochondrial antigens in vitro. Peripheral blood mononuclear cells were collected by lymphapheresis and cultured with or without pokeweed mitogen for 10 days. The culture supernatants were then tested for antibodies to mitochondrial antigens by both immunofluorescence microscopy and a microtiter ELISA. Peripheral blood mononuclear cells from 5 of 7 patients with primary biliary cirrhosis but from none of the healthy controls produced antibodies to mitochondrial antigens spontaneously (without pokeweed mitogen stimulation). In contrast, peripheral blood mononuclear cells from 6 of 7 patients with primary biliary cirrhosis and from 6 of 7 control subjects synthesized detectable levels of antibodies to mitochondrial antigens after stimulation with pokeweed mitogen. In general, peripheral blood mononuclear cells from the primary biliary cirrhosis patients produced higher titers of antibodies to mitochondrial antigens in culture than cells from healthy controls. Furthermore, the antibodies to mitochondrial antigens reactivity produced by peripheral blood mononuclear cells of primary biliary cirrhosis patients exhibited a specificity for the M2 mitochondrial antigen which is present on the inner membrane of mitochondrial cristae and which is closely associated with a mitochondrial ATPase activity. In contrast, the antibodies to mitochondrial antigens reactivity produced by peripheral blood mononuclear cells of healthy controls appeared to be directed at a broader range of mitochondrial antigens. These findings indicate that, in patients with primary biliary cirrhosis, there is a marked expansion of B lymphocyte clones that produce an antibody to a specific mitochondrial antigen.
The streaming liver: A slow, divided flow?
Authors: Scothorne R. J. , Macsween R. N. M.
Published: March 2007

Twenty male adult rats weighing 200 g were injected with tritiated thymidine (3HTdR). The animals were then killed in groups of five, at the following times: 1 h, 1, 3 and 5 weeks. Autoradi-ograms of sections through the liver were prepared. The distances between labelled cells and the portal space rim were measured. One hour after labelling most labelled cells were confined to a region extending from the portal space rim up to a distance of 700 μm, which roughy corresponds to Rappaport's hepatic acinus zones-1 and -2. Throughout the experiment lasting 5 weeks labelled cells entered zone-3 and advanced toward the terminal hepatic vein. Hepatocytes travelled at a daily velocity of 1.44 μm, covering daily 0.324% of the acinus diameter. During the experiment acinus size did not change appreciably. The estimated mean hepatocyte cell cycle time was 37 days and its life expectation, 201 days. These experiments show that the liver is essentially a slowly renewing cell population. Hepatocytes nascent at the portal space gradually stream toward the terminal hepatic vein where they are probably eliminated by apoptosis. Their journey lasts 201 days. Since hepatocytes are glued together with tight junctions, all have to advance toward their terminal hepatic veins en masse. During their voyage, they traverse the three acinus zones, and since in each they produce different enzymes, each zone represents a differentiation state of the advancing cell. It is suggested further that the streaming hepatocyte carries with it its nerve supply and is accompanied by sinusoidal endothelium and Kupffer cells.
Nonsurgical intrahepatic portacaval shunt; a utopian dream or an approaching reality?
Authors: Rosch Josef
Published: March 2007

A prospective study was undertaken to assess the results of embolization of esophageal varices by the transjugular approach, in 83 patients with advanced cirrhosis (65 p. 100 Child's class C) admitted with severe bleeding (mean of 10 units of blood per patient). Embolization was successfully completed in 65 patients (78 p. 100) and complete obliteration was achieved in 50 (60 p. 100). Twenty-nine procedures were performed on actively bleeding patients. Cessation of bleeding was observed in 22 (76 p. 100). Excluding the 5 patients in whom embolization could not be achieved, the success rate was 92 p. 100. Embolization was attempted as an elective procedure in 54 patients who were not actively bleeding. Recurrent hemorrhage occurred in 18 patients (33 p. 100) during the first month of follow-up. Finally, 38 patients rebled and and 57 died. No accident could be ascribed to the procedure. It is concluded that: (a) the transjugular approach proved to be safe in actively bleeding patients with severe cirrhosis; (b) variceal obliteration effectively controlled active bleeding. The efficacy of this procedure was limited in time. The use of embolization is further hindered by a consistent failure rate and by the development of new therapeutic modalities, such as sclerotherapy.
Propranolol for the prevention of recurrent variceal hemorrhage: A controlled trial
Authors: Villeneuve Jean-Pierre , Pomier-Layrargues Gilles , Infante-Rivard Claire , Willems Bernard , Huet Michel P. , Marleau Denis , Viallet André
Published: March 2007

We conducted a prospective, randomized single-blind trial of propranolol for the prevention of recurrent variceal bleeding. Seventy-nine patients shown to have variceal hemorrhage at endoscopy were included in the study within 72 hr following diagnosis. Fifty-seven patients had alcoholic cirrhosis, 10 cryptogenic cirrhosis, 6 posthepatitic cirrhosis, 4 biliary cirrhosis, 1 portal vein thrombosis without cirrhosis and 1 idiopathic portal hypertension. The severity of liver disease at inclusion was assessed according to the Pugh modification of the Child-Turcotte classification: 9 (11%) had Class A; 41 (52%) Class B and 29 (37%) Class C disease. Patients were randomly assigned by sealed envelope to the propranolol group (42 patients) or the placebo group (37 patients). Propranolol dosage was titrated in order to produce plasma concentrations of propranolol of 50 to 150 ng per ml. β-blockade was also confirmed by isoproterenol testing. The cumulative percentages of patients free of rebleeding 1 and 2 years after inclusion were 31 and 21% in the propranolol group, and 25 and 17% in the placebo group; both differences were not significant. Cumulative 1 and 2 years survival were also comparable: 64 and 54% in the propranolol group vs. 70 and 63% in the placebo group. There was no evidence for a therapeutic effect of propranolol after adjusting for potential confounding variables by multiple logistic regression. We conclude that propranolol is not effective for the prevention of variceal rebleeding, when administered early following the initial bleed, in cirrhotics unselected with respect to the severity of the liver disease.
Hepatic α1-antitrypsin mRNA content in cirrhosis with normal and abnormal protease inhibitor phenotypes
Authors: Schwarzenberg Sarah Jane , Sharp Harvey L. , Manthei Robin D. , Seelig Steven
Published: March 2007

We quantitated α1-antitrypsin mRNA in normal, α1-antitrypsin-deficient cirrhotic and biliary cirrhotic livers using two-dimensional electrophoretograms of [35
S] methionine-labeled translational products of total hepatic RNA and RNA/DNA hybridization. α1-antitrypsin precursor product was identified by immunoprecipitation. The relative abundance of α1-antitrypsin product from normal (0.989 ± 0.197), cirrhotic (0.956 ± 0.062) and α1-antitrypsin deficient (0.818 ± 0.12) livers was not significantly different. Although (RNA/DNA) was decreased in the PiZZ cirrhotic livers compared to normal (0.56 ± 0.045 vs. 0.95 ± 0.225), it equaled that found in the PiM cirrhotic livers (0.56 ± 0.055). The concentration of α1-antitrypsin mRNA [relative abundance × (RNA/DNA)], while decreased in PiZZ compared to normal liver, is thus no different in PiZZ cirrhotics than in PiM cirrhotics. We confirmed this observation by quantitation of the α1-antitrypsin mRNA using an α1-antitrypsin genomic probe. By RNA/DNA hybridization, α1-antitrypsin mRNA was equal in PiM cirrhotic and PiZZ cirrhotic (38.48 ± 4.5 vs. 31.93 ± 2.1), but significantly decreased from noncirrhotic PiM liver (58.36 ± 12.7). We conclude that α1-antitrypsin mRNA is decreased in cirrhosis of any etiology, and this decrease appears to represent a general response of the liver to injury. Since the decreased α1-antitrypsin mRNA in PiM cirrhotics is associated with normal serum α1-antitrypsin levels, it is unlikely that the decreased α1-antitrypsin mRNA in PiZZ cirrhotics accounts for their decreased serum levels.