bluuebunny/crossref_metadata_2025 huggingface dataset:
Ultrastructural Evidence of Intrahepatic Cholestasis Before and After Chenodeoxycholic Acid Therapy in Patients With Cholelithiasis: the National Cooperative Gallstone Study
Authors: Phillips James M. , Fisher Rosemarie L. , Anderson Deborah W. , Lan Shu-Ping , Lachin John M. , Boyer James L.
Published: July 2010

Electron microscopy was performed to assess potential hepatotoxicity before and after 9 and 24 months of chenodeoxycholic acid (CDCA) therapy (375 or 750 mg, daily) in 103 patients with cholelithiasis. Prior to treatment, 64% of the biopsies demonstrated ultrastructural evidence of intrahepatic cholestasis, manifested by abnormal bile canaliculi, thickened pericanalicular ectoplasm, and retention of biliary material within liver cells or pericellular spaces. After 9 months of CDCA therapy, several changes (including increased free cytoplasmic “biliary pigment,” decreased canalicular microvilli, and increased pericanalicular ectoplasm) consistent with intrahepatic cholestasis became more prevalent (63% prior to therapy, 89% at 9 months, p < 0.01) regardless of CDCA dose. After 24 months of therapy, canalicular microvilli and pericanalicular ectoplasm continued to be abnormal (p < 0.01).

This study indicates that ultrastructural evidence of intrahepatic cholestasis is a common subclinical abnormality in patients with cholelithiasis which increases during CDCA therapy, changes which could represent either the natural history of cholelithiasis or CDCA toxicity. A drug effect is suggested by the lack of correlation of these abnormalities with the duration of cholelithiasis prior to CDCA treatment and their “increasing” prevalence after only 9 months of therapy. In addition, canalicular membrane lesions developed in two patients which were similar to lithocholate toxicity in animals. It is not known whether these abnormalities would result in clinically significant hepatoxicity if therapy is continued for longer than 24 months.
Hepatic Free Fatty Acids in Alcoholic Liver Disease and Morbid Obesity
Authors: Mavrelis Peter G. , Ammon Helmut V. , Gleysteen John J. , Komorowski Richard A. , Charaf Ursula K.
Published: July 2010

Alcoholic liver disease is characterized by the accumulation of fat and inflammatory changes in the liver. Because free fatty acids, the precursors of triglycerides, can damage biological membranes, accumulation of free fatty acids in the liver might be in part responsible for the functional and morphological changes seen in alcoholic liver disease. We, therefore, determined the hepatic lipid composition in biopsies from 31 patients with alcoholic liver disease, 18 patients with morbid obesity, and 5 patients without evidence of liver disease. Free fatty acids were found in all liver biopsies. Patients with morbid obesity or alcoholic liver disease had significantly higher fatty acid and triglyceride levels than did controls (p <, 0.01). Patients with alcoholic liver disease had significantly higher fatty acid levels than did patients with morbid obesity (p < 0.05), while there was no difference in the triglyceride concentrations between these two groups. The distribution of the fatty acids in the free fatty acid fraction differed significantly from that in the triglyceride fraction indicating a preferential incorporation of unsaturated fatty acids into triglycerides. This difference in the distribution pattern was lost in patients with the most severe forms of alcoholic liver disease. The data are consistent with the hypothesis that accumulation of free fatty acids in patients with alcoholic liver disease may be responsible for or contribute to the observed functional and morphological damages.
Galactosamine Hepatitis, Endotoxemia, and Lactulose
Authors: Van Vugt Hendrina , Van Gool Jacobus , Thomas Lambert L.M.
Published: July 2010

Studies by Liehr et al. suggest that endotoxins are important in the pathogenesis of galactosamine hepatitis (Gal–N hepatitis) in rats. Lactulose (9.1 gm per kg per day) prevents hepatic lesions induced by Gal–N; an antiendotoxin effect of lactulose is postulated. However, commercial preparations of lactulose are contaminated with galactose, which shows a competitive action to Gal–N. To analyze the effect of galactose, male Wistar rats were pretreated with lactulose (Duphalac®, 9.1 gm per kg per day) and given Gal–N (375 mg per kg i.p.). After 24 hr, serum was analyzed for glutamic pyruvate transaminase, glutamate dehydrogenase, and sorbitol dehydrogenase activities. Pretreatment with Duphalac®, even 1 hr before Gal–N, abolished toxicity. Duphalac® contains 10 gm galactose per 100 ml. Galactose was given in a similar concentration and similar inhibition occurred. Pretreatment with purified lactulose (9.1 gm per kg for 5 days) diminished the effects of Gal–N but did not normalize enzyme concentrations. Because small doses of galactose (80 and 300 mg per kg) showed similar inhibitory effects, we conclude that the protective effect of commercial lactulose preparations is mainly due to galactose contamination and not to an antiendotoxin effect.
Differential Effects of Oral Contraceptive Steroids on the Metabolism of Benzodiazepines
Authors: Patwardhan Rashmi V. , Mitchell Mack C. , Johnson Raymond F. , Schenker Steven
Published: July 2010

The effects of oral contraceptive steroids (OCS) on the disposition and elimination of lorazepam, oxazepam, and chlordiazepoxide were examined. Lorazepam and oxazepam are metabolized via glucuronidation while chlordiazepoxide is metabolized by oxidation in the liver. The disposition and elimination of lorazepam, oxazepam, and chlordiazepoxide was studied in females not taking OCS and females taking OCS (norethindrone acetate, 1 mg; ethinyl estradiol, 50 m¨g) for 6 months or more. The t½ (β) for lorazepam was significantly reduced in women taking OCS (6.0 ± 3.1 vs. 14.0 ± 6.2 hr) (p < 0.005) as compared to controls, and the tV4 (/}) for oxazepam was reduced in women taking OCS (7.71 ± 3.23 vs. 12.09 ± 5.08 hr) as compared to controls, but did not reach statistical significance. The plasma clearance of both lorazepam and oxazepam was significantly increased in women taking OCS [(288.9 ± 165.9 vs. 77.5 ± 3.29 ml per min) (p < 0.01) and (251.2 ± 106.9 vs. 97.86 ± 69.4 ml per min) (p < 0.01), respectively] as compared to controls. The volumes of distribution of lorazepam and oxazepam were significantly increased in women taking OCS (p < 0.05) while plasma binding of these drugs was similar in both groups. In contrast, the t½ (β?) of chlordiazepoxide was significantly prolonged (20.58 ± 8.08 vs. 11.63 ± 5.91 hr) (p < 0.05), and the plasma clearance was significantly reduced (13.41 ± 4.69 vs. 33.22 ± 12.37 ml per min) (p < 0.05) in the OCS group as compared to controls. The volumes of distribution of chlordiazepoxide were similar in both groups, and the plasma binding of chlordiazepoxide tended to be lower in the OCS group but did not reach statistical significance. We conclude that OCS exert a differential effect on the elimination of benzodiazepines, whereby oxidation of chlordiazepoxide is impaired while the glucuronidation of lorazepam and oxazepam is enhanced by OCS.
Detection of Hepatitis B Virus DNA in Serum by a Simple Spot Hybridization Technique: Comparison with Results for Other Viral Markers
Authors: Scotto Jacques , Hadchouel Michelle , Hery Christiane , Yvart Jeannine , Tiollais Pierre , Brechot Christian
Published: July 2010

A simplified spot method for determination in serum of hepatitis B virus DNA (HBV DNA) by molecular hybridization is proposed. For simultaneous testing of 30 serum samples, it reduced to about 1 hr the duration of the steps preceding hybridization proper. The method also greatly reduced the loss of DNA during these steps and allowed more sensitive detection in samples of only 25 or 50 μ1.

HBV DNA was determined in 181 serum samples by this method, and the results were pooled with 67 previous determinations by the Southern blot technique. Results for the pool were then compared to those obtained with radioimmunoassay for serological HBV markers.

Ninety-six of the 248 samples were HBV DNA positive. Eleven others gave variable or inconclusive results, probably due to low viral particle titers.

Seventy-two HBsAg- and HBeAg-positive sera contained HBV DNA, confirming that HBeAg is a marker of active viral replication. Fourteen other HBsAg- and HBeAg-positive sera, obtained from eight patients, were either HBV DNA negative or oscillated between negative and positive, or, again, were weakly positive; serological follow-up in 7 patients showed seroconversion to anti-HBe in 5, 3 of which became HBsAg negative. Eight of the HBsAg-positive sera were negative or borderline for HBeAg but contained HBV DNA and may, therefore, have been infective; seven of these sera had anti-HBe. Six HBsAg-negative sera contained HBV DNA and may also have been infective; five of these exhibited HBV antibodies.

These results indicate that molecular hybridization not only provides a more sensitive and direct method for detecting hepatitis B virus in serum but also defines additional serological patterns with predictive or epidemiological value.
Cytoskeleton Antibodies in Chronic Active Hepatitis, Primary Biliary Cirrhosis, and Alcoholic Liver Disease
Authors: Kurki P. , Miettinen A. , Salaspuro M. , Virtanen I. , Stenman S.
Published: July 2010

Antibodies to cytoplasmic microfilaments, intermediate filaments (vimentin filaments), and microtubules which comprise the cytoskeleton of the cell were assayed in sera from 23 patients with HBsAg-negative chronic active hepatitis and 1 with HBsAg-positive chronic active hepatitis, 15 patients with primary biliary cirrhosis, 20 patients with alcoholic liver disease, and 32 healthy controls. The cytoskeleton antibodies were assayed by indirect immunofluorescence technique using vinblastine-treated cultured human embryonic fibroblasts as substrates.

There was a significantly increased incidence of cytoskeleton antibodies in patients with liver disease as compared to the control group. Antibodies to microfilaments were found frequently in sera from patients with chronic active hepatitis (67%) and primary biliary cirrhosis (53%) but were rare in sera from patients with alcoholic liver disease (25%) and in control sera (3%). On the other hand, antibodies to microtubules were found in 50% of sera from patients with alcoholic liver disease but in only 7 to 13% of sera from other groups. Intermediate filament antibodies of IgG or IgA class were found only in patient sera whereas intermediate filament antibodies of IgM class were found in the majority of sera in all groups including control sera. The highest titers of intermediate filament antibodies were seen in primary biliary cirrhosis and chronic active hepatitis. The production of cytoskeleton antibodies may be due to the reorganization or destruction of cytoskeletal structures in the liver.
Evidence for the Hormone Dependency of Hepatic Hyperplastic Nodules: Inhibition of Malignant Transformation After Exogenous 17β-Estradiol and Tamoxifen† ‡
Authors: Mishkin Shaindel Y. , Farber Emmanuel , Ho Ru Kun , Mulay Shree , Mishkin Seymour
Published: July 2010

Hepatic hyperplastic nodules (HHNs) in rats were studied as an experimental prototype of oral contraceptive-related hepatic tumors. We have found cytoplasmic estrogen receptors in HHNs produced by acetylaminofluorene (AAF) (four cycles of 0.02% in diet). Rats with AAF-induced HHNs were randomized into four groups: (i) AAF-treated control; (ii) estrogen alone (estradiol-17β); (iii) tamoxifen alone, and (iv) estrogen + tamoxifen. After 8 months of treatment with estrogen (estradiol-17β) in combination with tamoxifen, there was regression of nodular involvement and no evidence of malignant transformation. Decreased nodular proliferation also occurred after 2 and 4 months treatment with estradiol-17β and after 8 months of tamoxifen administration. The incidence of hepatocellular carcinoma after 8 months of treatment was significantly less after treatment with estrogen (40%) or tamoxifen (42.9%) when compared to AAF-treated controls (87.5%). The number of -γ-glutamyitranspeptidase-positive foci were reduced in all treatment groups after 2,4, and 8 months of treatment; these changes were most pronounced in the estrogen-treated group and did not directly correlate with the per cent inhibition of malignant transformation. Our results suggest that the malignant transformation of estrogen receptor-positive HHNs is hormone dependent.
Development and Use of a Rat Albumin cDNA Clone to Evaluate the Effect of Chronic Ethanol Administration on Hepatic Protein Synthesis†
Authors: Zern Mark A. , Chakraborty Prasanta R. , Ruiz-Opazo Nelson , Yap Sing H. , Shafritz David A.
Published: July 2010

A rat albumin cDNA probe (pBR alb 149) was developed in order to investigate the molecular mechanisms responsible for changes in hepatic protein synthesis after chronic administration of ethanol to rats. Rats fed a diet for up to 1 year in which 36% of calories were from ethanol, developed fatty livers but not cirrhosis. Cell-free protein synthesis with liver membrane-bound polysomes of ethanol-fed rats was increased as compared to control membrane-bound polysomes, whereas protein synthesis with free polysomes was unchanged. Total RNA extracted from liver membrane-bound polysomes and translated in a rabbit reticulocyte mRNA-dependent system showed a marked increase in albumin synthesis in the ethanol-fed group. Analysis of RNA molecules separated according to molecular weight by gel electrophoresis and hybridized with recombinant-cloned albumin cDNA demonstrated an increase in full-sized albumin mRNA species in ethanol-fed animals. Therefore, chronic ethanol administration appears to increase albumin synthesis by increasing the steady-state level of biologically active albumin mRNA in liver membrane-bound polysomes. Despite development of fatty liver, the protein synthesis machinery functions normally.
Ethanol Effects in a Rat Hepatoma Cell Line: Induction of γ -Glutamytransferase
Authors: Barouki Robert , Chobert Marie-Noële , Finidori Joëlle , Aggerbeck Martine , Nalpas Bertrand , Hanoune Jacques
Published: July 2010

The clone C2 derived from a rat hepatoma cell line was used to investigate the mechanism of the induction of γ-glutamyltransferase by ethanol. γ-glutamyltransferase activity was detected in the C2 cell (1.4 mU per mg protein), and its kinetic properties were similar to normal rat liver γ-glutamyltransferase. Ethanol provoked a dose- and time-dependent increase in γ-glutamyltransferase activity, the maximum (2- to 3-fold) occurring 48 hr after the addition of ethanol (180 mM). In contrast, the activity of five other enzymes tested were not markedly modified by ethanol. Propanol was more potent than ethanol in inducing 7-glutamyltransferase (5-fold stimulation), whereas methanol had no effect. The release of the enzyme in the medium was increased by ethanol and propanol.

Several observations argue in favor of an increase in the biosynthesis of γ-glutamyltransferase after ethanol addition: (i) ethanol increased the maximal velocity of the enzyme and did not modify the affinity for its substrates. It did not alter γ-glutamyltransferase subcellular distribution; (ii) ethanol had no immediate effect when added directly to the assay mixture; (iii) the lag period and the time course of the increase in γ-glutamyltransferase activity were those expected for an induction process; (iv) the increase in γ-glutamyltransferase activity was prevented by cyclohex-imide and actinomycin D suggesting that ethanol acted at the transcriptional level. The effect of ethanol was not mimicked by acetaldehyde. In conclusion, we have demonstrated that ethanol increases the biosynthesis of γ-glutamyltransferase in a rat hepatoma cell line which provides a new in vitro system.
Natural History of Hepatitis B Virus Infection in Renal Transplant Recipients-A Fifteen-Year Follow-Up
Authors: Dusheiko Geoffrey , Song Ernest , Bowyer Sheila , Whitcutt Michael , Maier Gunthild , Meyers Anthony , Kew Mlchael C.
Published: July 2010

Hepatitis B virus (HBV) markers were measured in 83 immunosuppressed renal transplant patients who were followed for periods of 2 to 15 years. Sixty-nine patients were negative for HBsAg before transplantation, of whom 14 were positive for anti-HBs. The remaining 14 patients were HBsAg positive prior to transplantation. Eighteen patients were identified as being HBsAg positive during the follow-up period. Four patients acquired primary type B hepatitis; one died of submassive hepatic necrosis and the remaining three became chronic HBV carriers with positive HBeAg, DNA polymerase, and HBV DNA. Several patterns of HBV expression were observed in HBsAg-positive patients. Four patients were HBsAg, HBeAg, DNA polymerase, and HBV DNA positive prior to transplantation, and these markers persisted. Reactivation of HBV replication occurred in eight patients, seven of whom were HBsAg positive and HBeAg and anti-HBe negative originally; one patient was anti-HBc positive. A single patient was HBsAg and anti-HBe positive and remained so for 22 months. The remaining previously HBsAg-positive patient is currently HBsAg negative. These serological data suggest that reactivation of HBV replication or continued hepatitis B virion replication occurs as commonly or more commonly than de novo infection in renal transplant recipients. The presence of HBeAg in serum predisposes to long-term Dane particle expression in immunosuppressed patients, whereas anti-HBe-positive carriers may not always be susceptible to reactivation of HBV replication despite immunosuppression.
Measurement of Serum α1-Acid Glycoprotein and cei-Antitrypsin Desialylation in Liver Disease
Authors: Serbource-Goguel Nathalie , Corbic Michèle , Erlinger Serge , Durand Geneviève , Agneray Jean , Feger Jeanne
Published: July 2010

To determine whether the presence of circulating desialylated glycoproteins reflect the existence and/or the severity of liver disease, 73 patients were evaluated with liver biopsies, conventional liver function tests, and the measurement of the degree of desialylation of two glycoproteins ax-acid glycoprotein (α1-AGP) and ai-antitrypsin (α1-AT). A combination of two immunological methods, available as routine laboratory tests, was used for the determination of the desialylation of α1-AGP and α1-AT. The severity of liver disease was assessed by a clinical classification depending upon the presence or absence of four complications (jaundice, ascites, hepatic encephalopathy, and weight loss). The presence of serum desialylated α1-AGP did not allow detection of mild liver disease, but asialoon-AGP (and to a lesser extent of asialo-ai-(AT) correlated with the severity of liver disease. The sensitivity of desialylated ai-AGP in detection of severe liver disease was 65%, and its specificity was 80%.
Increased Synthesis of Rat Liver Alkaline Phosphatase by Bile Duct Ligation
Authors: Kaplan Marshall M. , Ohkubo Akiyuki , Quaroni Elaine G. , Sze-Tu Duncan
Published: July 2010

The following studies were done to determine the mechanism of the increse in rat liver alkaline phosphatase activity after bile duct ligation. Antiserum was raised in rabbits to highly purified rat liver alkaline phosphatase. In immune titration experiments, the 350% increase in rat liver alkaline phosphatase activity caused by bile duct ligation was paralleled by a similar increase in immuno-precipitated alkaline phosphatase protein. In a second set of experiments, rat liver alkaline phosphatase was labeled with L-[3H]leucine injected into portal veins. Alkaline phosphatase was purified by antibody affinity chromatography followed by immunoprecipitation. The incorporation of L-[3H]leucine into alkaline phosphatase was significantly higher in bile duct-ligated rats than in controls, 68,357 ± 7,144 vs. 19,297 ± 3,076 dpm per gm liver (p < 0.001) and 349 ± 36 vs. 104 ±17 dpm per gm protein (p < 0.001). In a third set of experiments, the incorporation of L-3H-amino acid into highly purified rat liver alkaline phosphatase was measured. Rat liver alkaline phosphatase was purified by means of sequential iV-butanol extraction, antibody affinity column chromatography, preparative polyacrylamide gel electrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. There was significantly more L-3H-amino acid incorporated into alkaline phosphatase in bile duct-ligated rats compared to sham-operated rats, 3,565 vs. 704 dpm per gm liver and 19,656 vs. 3,843 dpm/gm protein. The data suggest that bile duct ligation increases the synthesis of rat liver alkaline phosphatase.
Nervous and Ocular Disorders in Children with Cholestasis and Vitamin A and E Deficiencies
Authors: Alvarez Fernando , Landrieu Pierre , Laget Paul , Lemonnier Frédérique , Odièvre Michel , Alagille Daniel
Published: July 2010

Thirteen children, aged 10 months to 20 years, presenting with chronic cholestasis from the first month of life and with low serum levels of vitamins A and/or E, have been investigated for neurological and ophthalmological symptoms. Clinical findings consisted of 4 types: peripheral neuropathy; cerebellar dysfunction; abnormalities of eye movement, and retinal degenerative changes. The results of electrophysiological and morphological studies of muscle and nerves were consistent with jieurono-axonal degeneration. Electrical abnormalities of the retina, especially a decrease of the b wave of electroretinogram, appear to be the first sign of the syndrome, allowing early detection. Evidence for vitamin deficiency (E or E+A) suggests substitutive parenteral treatment in such patients.
Abnormal Glucuronidation of Zomepirac in Patients with Cirrhosis of the Liver
Authors: Witassek Felix , Bircher Johannes , Huguenin Philipp , Preisig Rudolf
Published: July 2010

The disposition of zomepirac was investigated in 18 patients with various liver diseases and in 10 healthy normal subjects in order to further test the hypothesis that glucuronidation of drugs may be spared in liver disease. Severity of the liver disease was assessed by the galactose elimination capacity. Following oral administration of zomepirac (200 mg), plasma and urinary drug concentrations were measured by high-pressure liquid chromatography. Urine was assayed before and after alkaline hydrolysis. The apparent oral clearance of zomepirac was 3.7 ± S.D. 1.2, 3.0 ± 0.8, and 1.8 ± 0.6 mlmin_1kg_1in normal subjects, patients with mild liver disease, and cases with cirrhosis, respectively. In patients with liver disease, the reduction in zomepirac clearance was significantly correlated with the abnormalities in galactose elimination capacity (r = 0.83, n = 18), suggesting that the functioning liver cell mass was the major determinant of the rate of zomepirac disposition. These results are not consistent with the original hypothesis but suggest that-in contrast to ether-glucuronidation-ester-glucuronidation may be abnormal in liver disease. Dosage adjustments may, therefore, be necessary in patients with cirrhosis of the liver.
Massive Blastic Infiltration of the Liver: A Cause of Fulminant Hepatic Failure
Authors: Zafrani Elie S. , Leclercq Bernard , Vernant Jean-Paul , Pinaudeau Yvon , Chomette Guy , Dhumeaux Daniel
Published: July 2010

The clinical and pathological findings in four cases of fulminant hepatic failure due to massive infiltration of the liver by acute leukemia or lymphoma are reported. Liver abnormalities were found simultaneously with or led to the discovery of hematologic malignancies, and consisted of marked hepatomegaly and severe hepatocellular insufficiency associated with hyperlactatemia. The blood malignancies were peculiar in their fast cellular growth and large tumor mass. Evolution was rapidly fatal in all these cases. In another patient, marked hepatomegaly and hyperlactatemia revealed the presence of a widespread lymphoma before the appearance of hepatocellular insufficiency. Immediate chemotherapy was instituted, and complete remission without hepatic complication was obtained. It is suggested that malignant hematological diseases with fast cellular growth may present as fulminant hepatic failure. In order to avoid a rapidly fatal outcome secondary to liver failure and metabolic disorders, early recognition of these malignancies is necessary so as to assure prompt administration of appropriate chemotherapy.
Hepatitis B Virus Infection: The Impact of Molecular Biology
Authors: Sherlock Sheila , THomas Howard C.
Published: July 2010

Assays for the detection of nucleic acid in serum are likely to be better systems for determining infectivity than indirect ones dependent on detection of virus-encoded proteins such as HBeAg. The hybridization assays are particularly useful in monitoring spontaneous or treatment-related conversion from the “replicative” to “nonreplicative” phase of HB V infection. The simplified technology described in the papers reported in this issue of Hepatology (9, 14) and their future adaptation to nonisotopic systems will rapidly bring these techniques into everyday clinical practice.
Hepatic Thyroid Hormone Levels Following Chronic Alcohol Consumption: Direct Experimental Evidence in Rats Against the Existence of a Hyperthyroid Hepatic State
Authors: Teschke Rolf , Moreno Fernando , Heinen Edgar , Herrmann Jorg , Kruskemper Hans-Ludwig , Strohmeyer Georg
Published: July 2010

To study the effect of chronic alcohol consumption on hepatic levels of thyroid hormones, female Sprague-Dawley rats (n = 24) were pair-fed nutritionally adequate liquid diets containing either ethanol (36% of total calories) or isocaloric carbohydrates for 21 days. Compared to controls, chronic alcohol consumption failed to result in a significant change of hepatic thyroid hormone levels (thyroxine: 14.7 ± 1.81 ng per gm of liver wet weight vs. 15.0 ± 1.59; triiodothyronine: 2.60 ± 0.16 ng per gm of liver wet weight vs. 2.66 ± 0.18). Similar results were obtained when the hepatic levels of thyroid hormones were expressed per total liver, per gram of liver protein or per 100 gm of body weight. Moreover, prolonged alcohol ingestion led to a significant reduction of serum total thyroxine by 31.6% (p ± 0.001), free thyroxine by 38.9% (p ± 0.02), total triiodothyronine by 40.2% (p ± 0.001) and free triiodothyronine by 56.1% (p ± 0.001) when compared to their pair-fed controls, whereas thyroid-stimulating hormone levels remained virtually unchanged. These data, therefore, clearly show that chronic alcohol consumption is incapable of creating a hyperthyroid hepatic state in rats, and limit the rationale for antithyroid treatment in patients with alcoholic liver disease.
Maintenance Cultures of Kupffer Cells Isolated from Rats of Various Ages: infrastructure, Enzyme Cytochemistry, and Endocytosis†
Authors: De Margreet Leeuw A. , Brouwer Adriaan , Barelds Roel J. , Knook Dick L.
Published: July 2010

Sinusoidal liver cells were isolated from the livers of 3-, 12-, 30-, and 36-month-old female BN/ BiRij rats by enzymatic digestion. The Kupffer cells in the sinusoidal eel suspensions were purified by centrifugal elutriation and kept in maintenance culture for periods of up to about 3 weeks. The viability and yield of Kupffer cells per gram of body weight did not change with the age of the donor rat. The ultrastructural, cytochemical, and functional characteristics of Kupffer cells as observed in perfusion-fixed liver were retained during several days of maintenance culture. The consistent observation of worm-like structures in cultured Kupffer cells indicated the reformation of the specific fuzzy coat of the cells during culture. Endogenous peroxidatic and acid phosphatase activities were evident in cultured Kupffer cells and showed the same localization as observed in perfusion-fixed liver. Kupffer cells in culture were able to endocytose colloidal carbon, latex particles (0.8 μm), horseradish peroxidase, and endotoxin, indicating the reappearance of different types of specific membrane receptors. The ultrastructural appearance of Kupffer cells was not markedly influenced by the age of the donor rat. However, with increasing age, the lysosomes showed increasing amounts of electron dense lipid-like material and iron in the form of ferritin. No qualitative age-related changes in the enzymes tested or in the endocytic capacity of the Kupffer cells were observed. On the basis of these observations, maintenance cultures of purified Kupffer cells can be considered as a valuable model for studying Kupffer cell functions, also in relation to aging phenomena.
bolism of the Inhibitory Neurotransmitter γ-Aminobutyric Acid in a Rabbit Model of Fulminant Hepatic Failure
Authors: Ferenci Peter , Covell David , Schafer Daniel F. , Waggoner Jeanne G. , Shrager Richard , Jones Anthony E.
Published: July 2010

The serum concentration of 7-aminobutyric acid (GABA) has been found to be elevated by more than an order of magnitude in a rabbit model of acute hepatic failure. A potential mechanism for increased serum GABA levels in liver failure is decreased catabolism of GABA. To study this possibility, GABA-transaminase activities were measured in nonneural tissues, and the kinetics of3H-GABAin vivowere defined in both normal rabbits and rabbits with galactosamine-induced acute liver failure. Liver failure was associated with 56 and 80% increases in the activities of GABA-transaminase in liver and kidney, respectively. In the normal rabbit following the intravenous injection of3H-GABA, total radioactivity in the serum decreased very rapidly initially and much more slowly after 3 to 4 min. Levels of labeled metabolites of3H-GABA increased very rapidly in serum and reached a plateau after 5 to 10 min. The dependence of total serum radioactivity on time was similar for rabbits 10 hr after galactosamine injection, and control rabbits, with the exception that total serum radioactivity was higher during the first 3 min of the study in galactosamine-treated rabbits. In rabbits in hepatic coma, total serum radioactivity was higher but radioactivity in metabolites was lower than in controls at corresponding times. Clearance of3H-GABA from the circulation (i.e., the product of the initial volume of distribution and the fractional catabolic rate, milliliters per minute) was reduced by 26% in rabbits 10 hr after galactosamine injection and by 61% in rabbits in hepatic coma. The reduction in clearance of3H-GABA was associated with a decrease in the initial volume of distribution but no appreciable change in the fractional catabolic rate of GABA. The magnitude of the decrease in clearance did not fully account for the increase in serum GABA concentrations observed during acute liver failure. These findings suggest that, in addition to impaired GABA metabolism, an increase in the delivery rate of GABA to the circulation occurs in acute hepatic failure. Possible mechanisms for the latter phenomenon include increases in synthesis of GABA from glutamate in the liver and/or kidney, synthesis of GABA by gut bacteria permeability of the intestine and brain to GABA.
Hepatitis A Infection in Chronic Carriers of Hepatitis B Virus
Authors: Zachoval Reinhart , Roggendorf Michael , Deinhardt Friedrich
Published: July 2010

By routine screening for serologic markers of hepatitis A and B in patients with acute hepatitis, 30 chronic carriers of hepatitis B virus with serologic evidence of acute hepatitis A and two patients with simultaneous acute infection with hepatitis A virus and hepatitis B virus were detected.

For evaluation of clinical data, two major risk groups were distinguished. Nine patients were drug addicts and 17 were children and young adults from Mediterranean countries or southeast Asia.

During the acute phase of illness, serum bilirubin and SGPT levels did not differ from those in other patients with acute hepatitis A. In three patients for whom follow-up sera were available, HBsAg concentration decreased during the acute stage of hepatitis A.
Are Nuclear Particles Specific for Non-A, Non-B Hepatitis?
Authors: De Vos Rita , Vanstapel Marie J. , Desmyter Jan , De Wolf-Peeters Chris , De Groote Guy , Colaert Jan , Mortelmans Jan , De Groote Jan , Fevery Johan , Desmet Valeer
Published: July 2010

This study reports the findings of an electron microscopic search for so-called non-A, non-B nuclear particles in liver biopsies from patients with mainly chronic or prolonged liver disease and from chimpanzees. In patients without hepatitis B virus or acute hepatitis A virus serological markers, non-A, non-B-like nuclear particles were seen in hepatocytes in 28 of 31 cases of presumed non-A, non-B hepatitis, but also in 11 of 12 cases of liver disease not usually attributed to hepatitis viruses. They were also seen in 22 of 24 patients with HBsAg, in 3 of 3 patients with anti-HBc and no HBsAg, in 1 of 2 patients with hepatitis A, in a case of cytomegalovirus hepatitis, and in 16 of 19 patients whose serology was not available or inconclusive. The particles were present in 1 of 8 untreated HBsAg-negative chimpanzees and in 2 of 2 HBsAg-positive chimpanzees. They appeared in 4 of 4 chimpanzees developing non-A, non-B hepatitis following exposure to various inocula. Three patterns of particle aggregates were distinguished, all of which had been shown by others in non-A, non-B hepatitis. Dense aggregates were predominant, while others have shown intermediate aggregates more often; reasons for this difference could be technical. No pattern was specific for any condition. Either non-A, non-B-like nuclear particles, although associated with non-A, non-B hepatitis, are not specific for this condition, or non-A, non-B hepatitis viruses are extremely more common than is currently appreciated.
Spontaneous Bacterial Peritonitis: A Prospective Investigation in Predominantly Nonalcoholic Cirrhotic Patients
Authors: Giovambattista  , Simonetti Rosa Giovanna , Craxí Antonio , Di Piazza Sonia , Spanó Caterina , Pagliaro Luigi
Published: July 2010

Two hundred and twenty-four consecutive inpatients with cirrhosis, mostly of nonalcoholic etiology, and ascites were prospectively investigated on admission for spontaneous bacterial peritonitis (SBP) by culture, smear, and polymorphonuclear (PMN) counts of ascitic fluid. Positive cultures were found in 54 patients. A possible intraabdominal source of infection was apparent in five of them, all with clinically overt peritonitis (secondary peritonitis). No intraabdominal source of infection was identified in the remaining 49. These patients were classified into the following groups: (a) SBP, 27 patients (positive cultures plus PMN counts ± 250 per mm3). Nine of them had no clinical features of peritonitis (silent SBP) in contrast to the other 18 (symptomatic SBP). (b) Bacteriascites (BA), 18 patients (positive culture with PMN counts less than 250 per mm3 in the absence of clinical peritonitis). Four patients were unclassified. Among 170 patients with negative culture, six had an overt peritonitis plus a positive smear and PMNs ± 250 per mm3 into ascites and were considered as probable SBP. The remaining 164 patients were classified as sterile ascites. A single strain of Gram-negative bacteria (usually of enteric origin) was recovered frequently in patients with symptomatic or silent SBP (79%) whereas Gram-positive bacteria were more frequently found in patients with BA (61%, p < 0.01). In-hospital mortality showed a statistically significant difference among patients with SBP (48%) and those with BA (22%) or sterile ascites (18%) (p < 0.05 and p < 0.0005, respectively). We conclude that SBP is frequent event in nonalcoholic cirrhosis. Bacterial contamination of ascites may remain in balance or progress to overt peritonitis according to the pathogenicity of the agent. SBP carries a high mortality in comparison to sterile ascites or BA despite early antibiotic treatment.
The Role of the Urea Cycle and Polyamines in Albumin Synthesis†
Authors: Oratz Murray , Rothschild Marcus A. , Schreiber Sidney S. , Burks Alvin , Mongelli Joseph , Matarese Barbara
Published: July 2010

Albumin synthesis is stimulated by those amino acids which increase urea synthesis and membrane bound polysome aggregation. Ornithine, an amino acid not incorporated into protein and produced from arginine in the urea cycle, is an albumin-stimulating amino acid and is the precursor of the polyamines, and we have shown that the polyamine spermine promotes bound polysome aggregation. To test the concept that ureogenesis with its generation of ornithine might play a key role in albumin synthesis regulation via the polyamine pathway, isolated livers from fasted donors were perfused with ornithine, α-difluoromethyl ornithine (DFMO), and spermine. In control experiments, albumin synthesis was 13.4 ± 0.8 mg per 100 gm liver per hr and polysome aggregation was 47%. These were increased in the presence of ornithine (26.0 ± 2.6 mg and 59%); if the livers were preperfused with DFMO before the addition of ornithine, then the expected increase in albumin synthesis and polysome reaggregation did not occur (16.3 ± 1.4 mg and 47%). However, if spermine was present with DFMO during the preperfusion, then the addition of ornithine had the expected effect (albumin synthesis = 26.1 ±1.2 mg and polysome aggregation = 62%). This suggests that if the ornithine to putrescine pathway is blocked, ornithine does not stimulate albumin synthesis and offers support to the concepts that (a) ornithine stimulation of albumin production is via polyamine synthesis and (b) that the urea cycle plays a more important role in protein metabolism than simply the pathway for nitrogen disposal.
Detection of Regan Variant Type of Alkaline Phosphatase Isoenzyme in Liver Tissue of Indian Childhood Cirrhosis
Authors: Parekh S. R. , Patel B. D. , Damle S. R. , Thanki G. M. , Khutti D.
Published: July 2010

Alkaline phosphatase (ALP) isoenzyme composition was studied in sera and liver from patients with Indian childhood cirrhosis (ICC). A typical pattern consisting of a fast-moving anodal preliver band with slower moving diffuse liver and placental bands followed by an intestinal band was consistently observed in sera of patients in all stages of ICC and in pregnant mothers of index ICC patients. ICC liver ALP was relatively heat-stable and inhibited by L-phenylalanine and L-leucine. The isoenzyme also had similar immunological determinants to placental ALP and adult intestinal ALP isoenzymes. Total serum ALP isoenzyme and its heat-stable component progressively increased in concentration from early to advanced stages of the disease suggesting that the diseased liver in ICC is the source of the abnormal isoenzyme.
Morphologic and Biochemical Changes in Dogs After Portacaval Shunt Plus Bile Fistula or Ileal Bypass: Failure of Bile Fistula or Ileal Bypass to Prevent Hepatocyte Atrophy
Authors: Thompson Jon S. , Porter Kendrick A. , Hayashida Nobuo , Mcnamara Donald J. , Parker Thomas S. , Russell William J. I. , Francavilla Antonio , Starzl Thomas E.
Published: July 2010

External biliary fistula (BF) or ileal bypass (IB) was performed in dogs at the time of or 2 weeks after portacaval shunt (PCS). The pathologic changes in the dog livers 2 to 4 weeks later were compared to those caused by PCS alone. Histopathologic differences between PCS alone vs. PCS plus BF or IB could not be found. Thus, the experiments did not confirm recent observations by others in rats that BF prevents or reverses the hepatic injury of PCS. As estimated by plasma mevalonic acid determinations, the increase in hepatic cholesterol synthesis that is characteristic after BF or IB was suppressed in animals with PCS. BF and IB reduced but did not eliminate the postprandial elevation in serum bile acid that occurs after PCS. The findings have possible relevance in planning the treatment of patients with familial hypercholesterolemia with the combined use of PCS and IB.
Non-A, Non-B Hepatitis: A Prospective Study of a Hemodialysis Outbreak with Evaluation of a Serologic Marker in Patients and Staff
Authors: Gitnick Gary , Weiss Steven , Overby Lacy R. , Ling Chung-Mei , Chairez Ruben , Parsa Khosrow
Published: July 2010

An outbreak of non-A, non-B hepatitis (NANBH) in a hemodialysis unit was prospectively studied and the clinical, biochemical, and serologic events were correlated with an experimental immunodiffusion assay for serum antigen and antibody. One hundred sixteen subjects (76 dialysis patients and 40 staff members) were studied over an 8-month period. Hepatitis was defined as two consecutive SGPT levels greater than two times the upper limit of normal occurring in two separate samples drawn greater than 7 days apart in the absence of other likely causes of liver disease. Weekly serum specimens were obtained and tested for SGPT, SGOT, alkaline phosphatase, bilirubin HBsAg, anti-HBc, anti-HBs, total anti-HAV, and anti-HAV IgM by commercial reagents, and for antigen and antibody by agar gel diffusion using reference reagents previously obtained from well-documented posttransfusion NANBH patients. Clinical evaluations were performed three times per week. Thirty patients and none of the staff developed NANBH. The NANBH patients were asymptomatic, except for two patients with jaundice. Fifteen of the 30 patients were positive for antigen which was detectable in at least one serum collected during the acute phase. Six patients and 10 staff without clinical NANBH or abnormal serology had antigen. Antigenemia was also observed in three patients with acute hepatitis B, with chronic hepatitis B in one patient and with alcoholic hepatitis in one patient. Thus, an antigen was detected in a high proportion of patients during the acute phase of NANBH, and it was also found in exposed patients who had other liver diseases. NANBH antigen was found in 19 of 72 (26.4%), transfused cancer patients without evidence of liver disease, and in 20% of 168 other control patients without liver diseases. Fifty per cent of patients with NANBH developed antigenemia compared to 13% of exposed patients (6 of 46) without NANBH (p < 0.05). Of eight patients who developed chronic hepatitis, six were NANB antigen positive. Transient antigenemia was found in some asymptomatic staff members whose liver tests remained normal. Immune serum globulin did not prevent transmission of NANBH under the circumstances of this study.
Transplantation of Woodchuck Hepatocellular Carcinoma in Nude Mice
Authors: Kobayashi Kenichi , Fukuoka Kenichi , Matsushita Fumiaki , Morimoto Hideo , Hinoue Yoshinobu , Honjo Hideo , Tanaka Nobuyoshi , Sugimoto Tatsuho , Kato Yasuhiro , Hattori Nobu , Ueda Shigeharu , Kato Shiro
Published: July 2010

Woodchuck hepatocellular carcinoma has been successfully transplanted into nude (athymic) mice. The morphology of heterotransplanted tumor is similar to that of naturally occurring hepatocellular carcinoma before transplantation. The growth rate of transplanted tumor was very slow compared with those of other transplanted tumors. During the first month, only two tumors appeared. However, definitive tumor growth was noted in 6 of 20 nude mice about 3 months later. Seventeen of 20 nude mice exhibited sustained tumor growth after 6 months. The woodchuck hepatocellular carcinoma in nude mice provides an in vivo model for the study of oncogenesis of human hepatocellular carcinoma related to hepatitis B virus.
Evidence For the Presence of the Asialoglycoprotein Receptor in Coated Vesicles Isolated from Rat Liver
Authors: Steer Clifford J. , Wall Doris A. , Ashwell Gilbert
Published: July 2010

Coated vesicles were isolated from rat liver by a modification of the procedure described by Nandi et al. for bovine brain (Proc. Natl. Acad. Sci. U.S.A. 1982; 79:5881-5885). The hepatic receptor for asialoglycoproteins was shown to be an integral constitutent of these vesicles as evidenced by their ability to bind 125I-asialo-orosomucoid in a specific and saturable manner. The specific binding activity of the purified vesicles was 17-fold greater than that of the original liver homogenate based on a protein determination. Binding was 97% latent and was made fully manifest only after removal of the clathrin coat and disruption of the exposed smooth membrane vesicles by detergent. Based on this finding, it was concluded that the binding protein for asialoglycoproteins was oriented toward the inner surface of the vesicle. In addition, evidence is presented that purification procedures employing detergent may result in coated vesicles deficient in one or more integral membrane proteins.
Vesicular Transport of Horseradish Peroxidase during Chronic Bile Duct Obstruction in the Rat†
Authors: Renston Richard H. , Zsigmond Gyula , Bernhoft Robin A. , Burwen Susan J. , Jones Albert L.
Published: July 2010

The vesicular transport system for biliary secretion of plasma-derived proteins was investigated in rats with chronic bile duct obstruction. Horseradish peroxidase, previously demonstrated to be a suitable tracer for vesicular transport, was employed in these studies. Both the time course of horseradish peroxidase secretion into bile and the morphological events in its uptake, transport, and biliary secretion were found to proceed in a manner essentially identical to that of sham-operated control animals. In addition, fragmentation of hepatocytes leading to sloughing into bile of large pieces of cytoplasm bearing horseradish peroxidase-containing endocytic transport vesicles frequently was observed in the cholestatic animals. These data suggest that the vesicular transport system for the secretion into bile of plasma-derived proteins remains intact and functional during chronic bile duct obstruction and that another mechanism, possibly fragmentation and solubilization of hepatocyte membranes followed by regurgitation of proteins released from endocytic vesicles, may be responsible for the elevation of biliary proteins within plasma seen during cholestasis.
A Prospective Randomized Study of Two Sclerotherapy Techniques for Esophageal Varices
Authors: Westaby David , Macdougall Brian R. D. , Melia Walter , Theodossi Andrew , Williams Roger
Published: July 2010

In a study designed to compare the efficacy and safety of two techniques of injection sclerotherapy, 40 patients (30 with cirrhosis and 10 with portal vein block) were randomly allocated to the sheath or free-hand technique. Although the former was associated with significantly less bleeding within the first 24 hr of injection (p < 0.05) but more postinjection pain (p < 0.05) and esophageal stricture, there was a trend toward earlier obliteration of varices. This was most marked over the first three courses of injection, and although frequency of rebleeding was not significantly less, none of the 11 episodes in the sheath group were fatal, compared to 5 of 15 bleeds in those injected by the free-hand technique (p < 0.05).
Ethanol Elimination in Males and Females: Relationship to Menstrual Cycle and Body Composition
Authors: Marshall Alex W. , Kingstone David , Boss Margot , Morgan Marsha Y.
Published: July 2010

Ethanol pharmacokinetics were determined following oral ethanol, 0.5 gm per kg, in nine normal women and 10 normal men, and related to total body water measured by 3H-water dilution and body fat determined anthropometrically. Ethanol pharmacokinetics were similar in the females throughout the menstrual cycle. No variation was seen in mean peak blood ethanol concentration or elimination rate in the midfollicular (Days 8 to 10) and midluteal (Days 22 to 24) phases. Mean peak blood ethanol values were significantly higher in females (88 ± 3 mg per 100 ml) than in males (75 ± 4 mg per 100 ml) (p < 0.05), and the mean area under the ethanol concentration-time curve was significantly greater in females (241 ± 12 mg hr per 100 ml) than in males (177 ± 11 mg hr per 100 ml) (p < 0.001). There was no significant sex difference in mean ethanol elimination rates. The mean apparent volume of distribution of ethanol in female (0.59 ± 0.02 liter per kg) was less than in males (0.73 ± 0.02 liter per kg) (p < 0.001). Both apparent volume of distribution of ethanol and area under curve were significantly correlated with total body water suggesting that the sex differences in ethanol pharmacokinetics were due to sex differences in body water content. The sex differences in ethanol pharmacokinetics may partly explain reports of male-female differences in the natural history of certain ethanol-related disorders.