bluuebunny/crossref_metadata_2025 huggingface dataset:
Hemagglutination and Immunofluorescence Studies on Polymerized Human Serum Albumin Binding Activity in Chronic Hepatitis B Virus Infection
Authors: Mora Ignacio , Porres Juan Carlos , Guio Carlos Hernández , Gutiez Julia , Carreño Vicente
Published: March 2007

The binding activity of polymerized human serum albumin was determined in 202 HBsAg carriers. The presence of polymerized human serum albumin receptor sites was tested by hemagglutination and differentiated from antihuman albumin antibodies by immunofluorescence, isolation of IgG and IgM fractions and testing of HBsAg anti–HBs immune complexes. A granular pattern with anti–HBs was specific for polymerized human serum albumin receptor sites as demonstrated with purified HBsAg. In addition, a linear pattern with fluoresceinated antihuman immunoglobulins might suggest the presence ofantihuman albumin antibodies (which was generally due to an IgG antibody). However, a granular pattern with fluoresceinated antihuman immunoglobulins may indicate the presence of HBsAg anti–HBs immune complexes. A weak linear pattern was also observed simultaneously in these cases, probably due to IgM antihuman albumin antibodies or an antipolymerized human serum albumin receptor site antibody.

Of 202 HBsAg–positive patients, 71 showed polymerized human serum albumin receptor sites activity. The highest percentage of polymerized human serum albumin receptor sites was found among patients showing HBeAg and hepatitis B virus DNA polymerase positivity (96%), followed by HBeAg positivity and hepatitis B virus DNA polymerase negativity (48%), and anti–HBe positivity and hepatitis B virus DNA polymerase negativity (17%). In addition, a significant correlation between polymerized human serum albumin titers and hepatitis B virus DNA polymerase was found (r = 0.573, p <0.01). However, at a similar HBeAg titer, patients who were positive for hepatitis B virus DNA polymerase had a higher polymerized human serum albumin receptor sites titer than those who were negative for hepatitis B virus DNA polymerase.

The presence of a granular pattern with anti–HBs or fluoresceinated antihuman immunoglobulins means a positivity to polymerized human serum albumin receptor sites or the presence of HBsAg anti–HBs immune complexes, respectively. Furthermore, polymerized human serum albumin receptor sites titers correlate with hepatitis B virus DNA polymerase concentrations and may serve as indicators of viral replication.
Effects of Ethanol on Experimental Hepatocarcinogenesis
Authors: Takada Akira , Nei Jinichi , Takase Shujiro , Matsuda Yoshiro
Published: March 2007

Although primary hepatoma is not very frequent in alcoholics, the incidence of hepatoma in cases of hepatitis B infection combined with heavy alcohol drinking is high. In the present study, the effects of chronic alcohol administration onthe development of chemicalinduced hepatic cancer in rats were analyzed. In 70% hepatectomized Wistar strain male rats, asingle dose (1 mg per 100 gm body weight) of diethylnitrosamine was injected intraperitoneally. Eight weeks after the injection, 20% alcohol–10% sucrose solution (diethylnitrosamine–alcohol group), 0.1% sodium phenobarbital solution (diethylnitrosamine–phenobarbital group), 10% sucrose solution (diethylnitrosamine–sucrose group) or tap water (diethylnitrosamine–alone group) was given as drinking water for 32 weeks. The numbers of visible nodules per liver were significantly greater in the diethylnitrosamine–alcohol and diethylnitrosamine–phenobarbital groups compared to the diethylnitrosaminealone and diethylnitrosamine–sucrose groups. The numbers of enzyme–altered foci which were positive to γ–glutamyl transpeptidase staining per square centimeter of liver section were also greater in the diethylnitrosamine–alcohol and diethylnitrosamine–phenobarbital groups than in the diethylnitrosamine–alone and diethylnitrosamine–sucrose groups, although the numbers of nodules and enzyme–altered foci were significantly larger in the diethylnitrosamine–phenobarbital group than in the diethylnitrosamine–alcohol group. The enzyme–altered foci areas calculated by γ–glutamyl transpeptidase staining were significantly larger in the diethylnitrosamine–alcohol and diethylnitrosamine–phenobarbital groups than in the diethylnitrosamine–alone and diethylnitrosamine–sucrose groups. Histologically, visible nodules observed in diethylnitrosamine–phenobarbital and diethylnitrosamine–alcohol groups showed characteristic features of neoplastic nodules. These results indicate that alcohol has a promoter action on the development of chemically induced hepatic cancer like phenobarbital.
Isosorbide Dinitrate in Experimental Portal Hypertension: A Study of Factors That Modulate the Hemodynamic Response
Authors: Blei Andres T. , Gottstein Jeanne
Published: March 2007

Isosorbide dinitrate, a long–acting vasodilator, has been tested in human portal hypertension with conflictingresults. In order to determine some of the factors that could affect the individual response to this drug, we infused isosorbide dinitrate at a low dose (10 to 25 μg per kg per min) and a high dose (100 μg per kg per min) to rats with portal vein stenosis. Under pentobarbital anesthesia, portal pressure was measured with an ileocolic vein catheter while cardiac output and regional blood flows were measured with the microsphere technique. At a dose that decreased arterial pressure by approximately 10%, cardiac output remained unchanged while portal vein inflow decreased significantly; portal pressure was not reduced (10.7 ± 0.2 vs. 10.0 ± 0.3 mm Hg), indicating a rise in portal vascular resistance. At a high dose of isosorbide dinitrate, arterial pressure and cardiac output fell markedly; portal pressure decreased only modestly (11.3 ± 0.3 vs. 9.8 ± 0.6 mm Hg, p < 0.05), but portal flow was unchanged, indicating a reduction in portal vascular resistance. In addition, portal hypertensive rats received a constant i.v. infusion of N–acetyl–cysteine the combination of the latter and isosorbide dinitrate markedly potentiated the effects on arterial pressure. Thus, the dose of the drug and the presence of cysteine–containing compounds appear to modulate the hemodynamic response to isosorbide dinitrate. Clinical testing with this drug should be undertaken with consideration of these factors.
The Usefulness of Microscopic Bile Examination in Patients With Suspected Microlithiasis: A Prospective Evaluation
Authors: Delchier Jean-Charles , Benfredj Paul , Preaux Anne-Marie , Metreau Jean-Michel , Dhumeaux Daniel
Published: March 2007

Gallbladder bile collected by duodenal intubation or during surgery was examined microscopically in patients who werefree of stones and in patients with proven stones. None of the 16 patients free of stones had cholesterol monohydrate crystals or calcium bilirubinate granules in bile. Among the 17 patients with proven cholelithiasis, 13 with cholesterol stones had cholesterol monohydrate crystals in their bile, but only 2 of the 4 patients with pigment stones had calcium bilirubinate granules. These data confirm that cholesterol monohydrate crystals are sensitive and specific for cholesterol stones, whereas calcium bilirubinate granules lack sensitivity for the diagnosis of pigment stones. From these results, thediagnostic usefulness of microscopic examination of bile collected from the duodenum was studied prospectively in 46 patients with symptoms suggestive of cholelithiasis but in whom stones had not been visualized at cholecystography and ultrasonography. In 15 of them, bile was found to be abnormal: cholesterol monohydrate crystals were seen in 11, cholesterol monohydrate crystals + calcium bilirubinate granules in 2 and calcium bilirubinate granules in 2. To date, nine of these patients have been operated on: 6 (all with cholesterol monohydrate crystals) had small cholesterol gallstones and 3 (2 with cholesterol monohydrate crystals and 1 with calcium bilirubinate granules) had signs strongly suggestive of the recent migration of gallstones. One patient refused operation, but minute pigment stones were found to be associated with calcium bilirubinate granules at duodenal intubation. In the other 31 patients, bile contained neither cholesterol monohydrate crystals nor calcium bilirubinate granules. They were not operated on and were followed up with repeated investigations for 12 to 24 months. During this period, cholelithiasis could be demonstrated in only one patient, who had pigment stones. These results suggest that microscopic bile examination can distinguish between patients with and without gallstones, and may be helpful in predicting the presence of cholelithiasis, when stones are not visualized by conventional procedures.
Clinical, Virologic and Histologic Outcome Following Seroconversion From Hbeag to Anti–Hbe in Chronic Hepatitis Type B
Authors: Fattovich Giovanna , Rugge Massimo , Brollo Lucio , Pontisso Patrizia , Noventa Franco , Guido Maria , Alberti Alfredo , Realdi Giuseppe
Published: March 2007

Seventy consecutive HBsAg– and HBeAg–positive patients with biopsy–proven chronic hepatitis were followed prospectively with serial determinations of SGPT levels and hepatitis B virus serum markers including HBsAg, HBeAg, anti–HBeand hepatitis B virus DNA. During a period of 1 to 11 years (mean ± S.D.: 5.0 ± 2.3 years), 28 patients remained persistently HBeAg positive, most with continuing biochemical and histologic activity, while 41 cases seroconverted to anti–HBe. One patient became HBeAg and anti–HBe negative. After seroconversion, 87.8% ofthe cases showed sustained normalization of SGPT, and clearance of hepatitis B virus DNA from serum and histologic improvement was documented in 79% of the cases who had a control liver biopsy, while 15.8% developed cirrhosis. In two patients (4.9%), the disease remained active despite seroconversion, and both cases had evidence of continuing hepatitis B virus replication. Finally, reactivation of liver damage and of hepatitis B virus replication was observed in three additional patients (7.3%) who had transiently normalized SGPT after seroconversion. All 70 patients were analyzed for hepatitis delta virus markers, and only two persistently HBeAg–positive cases were found positivefor antibody to hepatitis delta virus in serum, one also having hepatitis delta antigenin the liver. These findings indicate that, in chronic hepatitis type B, termination ofvirus replication is associated in most patients with biochemical and histologic regression of inflammatory activity. After anti–HBe seroconversion has occurred, virusreplication and liver disease may persist or reactivate in a small proportion of patients thus giving origin to the well–recognized group of anti–HBe positive, hepatitis B virus DNA–positive chronic hepatitis type B.
Effects of Immunosuppressive Therapy With Prednisolone on B and T Lymphocyte Function in Patients With Chronic Type B Hepatitis
Authors: Hanson Reginald G. , Peters Marion G. , Hoofnagle Jay H.
Published: March 2007

B and T lymphocyte function was studied in 10 patients with chronic type B hepatitis before, during and after a 28–day course of prednisolone therapy. Lymphocyte function was assessed by measuring the in vitro synthesis of immunoglobulin by peripheral blood mononuclear cells stimulated with pokeweed mitogen and by assaying lymphocyte proliferation in response to B and T cell mitogens. During high dose prednisolonetherapy, there was a decrease in immunoglobulin synthesis by peripheral blood mononuclear cells and in lymphocyte proliferation to all mitogens. Studies using separated B and Tcells showed that prednisolone treatment led to a decrease in both helper and suppressor T cell function but an enhancement of primary B cell function. When prednisolone was withdrawn, lymphocyte function rapidly returned to baseline levels.

During prednisolone therapy, serum aminotransferase activities decreased by an average of 50%. In all patients, there was a subsequent rebound increase in serum aminotransferase activities 4 to 10 weeks after withdrawal of prednisolone. This was accompanied by a striking increase in suppressor T lymphocyte activity without significant changes in either helper T cell or B cell function. The close correlation between changes in helper and suppressor T lymphocyte function and serum aminotransferase activities during andafter immunosuppressive therapy suggests that immunoregulatory T lymphocytes may play an important role in the pathogenesis of chronic type B hepatitis.
Products of the “X” Gene in Hepatitis B and Related Viruses
Authors: Feitelson Mark A.
Published: March 2007

The X region in hepatitis B virus DNA potentially encodes a polypeptide 154 amino acids in length. Two synthetic peptides spanning residues 100 to 115 (peptide 99) and 115 to 131 (peptide 100) in a hydrophilic domain within the carboxy terminal third of theproposed gene product were made and used to raise peptide antisera in rabbits. Such antisera specifically bound to X reactive determinants in liver–derived core antigen particles from humans (HBcAg), ducks (DHBcAg), ground squirrels (GSHcAg) and woodchucks (WHcAg) at each step of core antigen purification. This reactivity was blocked by addition of excess synthetic peptide, and neither sera were reactive with other purified antigens such as HBsAg. Individual polypeptides associated with these core particles were also reactive by Western blotting. These findings suggest that X reactive determinants are present in the core particles of hepatitis B virus and related viruses, and thatone or more coreassociated polypeptides may have both X and core antigenic determinants. The possible significance of these observations upon the genetic organization and expression of the X gene is discussed.
Value of Ascitic Lipids in the Differentiation Between Cirrhotic and Malignant Ascites
Authors: Jüngst Dieter , Gerbes Alexander L. , Martin Robert , Paumgartner Gustav
Published: March 2007

Ascitic fluid concentrations of cholesterol, triglycerides and phospholipids, were compared with ascitic fluid total protein in 40 patients with chronic liver disease, 51patients with various neoplasms and 1 patient with cardiac failure. Seven patients withboth chronic liver disease and malignancy were considered separately. The first 54 patients (23 cirrhotic and 31 with malignancy) were used to determine median values and ranges and to define the most suitable cutoff concentrations between both groups. Median values for cholesterol (75 mg per dl), phospholipids (0.79 mmole per liter), triglycerides (75 mg per dl) and protein (3.8 gm per dl)were higher in malignant ascites compared to ascitic fluid concentrations of cholesterol (20 mg per dl), phospholipids (0.33 mmole per liter), triglycerides (51 mg per dl) and protein (1.9 gm per dl) in patients withcirrhosis. The best discrimination values were 48 mg per dl for cholesterol, 0.6 mmole per liter for phospholipids, 65 mg per dl for triglycerides and 2.5 gm per dl for protein. Application of these cutoff points to 38 subsequent patients (17 cirrhotic, 1 with cardiac failure and 20 with malignancy) revealed an efficiency of 86.8% for cholesterol, 86.8% for phospholipids, 68.4% for triglycerides and 79.0% for protein. From the data of all 92 patients, an efficiency of 92.3% forcholesterol, 79.4% for phospholipids, 72.8% for triglycerides and 79.4% for protein was calculated.

We conclude that ascitic fluid cholesterol determination offers an excellent, cost–effective discrimination of ascites due to cirrhosis vs. ascites caused by malignancies.
Is the Acidity of Ascitic Fluid A Reliable Index in Making the Presumptive Diagnosis of Spontaneous Bacterial Peritonitis?
Authors: Pinzello Giovambattista , Virdone Roberto , Lojacono Francesca , Ciambra Maddalena , Dardanoni Gabriella , Fiorentino Germana , Riccobono Loredana , Pagliaro Luigi
Published: March 2007

Ascitic fluid pH and arterial–ascitic fluid pH gradient were compared to ascitic fluid polymorphonuclear cell count in 84 patients with cirrhotic ascites and in 12 with malignant ascites to assess their role as diagnostic tests for spontaneous bacterial peritonitis and to clarify the relationship between ascitic fluid pH and lactate. Ascitic fluid pH was significantly lower (pH 7.30) in spontaneous bacterial peritonitis (n = 18) and probable spontaneous bacterial peritonitis (n = 12) than in sterileascites (pH 7.41; n = 54). Since blood pH levels were not different in the presence of infection, arterial–ascitic fluid pH gradient was significantly higher in spontaneous bacterial peritonitis and probable spontaneous bacterial peritonitis than in sterile ascites (0.12 vs. 0.02). The close correlations between arterial–ascitic pH gradient and lactate (r = 0.77, p < 0.0001), lactate and bicarbonate gradient(r = 0.64, p = 0.003) and arterial–ascitic pH gradient and pCO2gradient (r = —0.90, p < 0.0001) suggest that the low ascitic fluid pH may be due to an increase in lactate and CO2. Patients withEscherichia coli–induced spontaneous bacterial peritonitis had significantly lower ascitic fluid pH and higher lactate than those with spontaneous bacterial peritonitisbyother organisms. Values of ascitic fluid pH, lactate and arterial–ascitic fluid pH gradient in malignant ascites were similar to those of spontaneous bacterial peritonitis and probable spontaneous bacterial peritonitis. Cutoff points, selected by receiver operating characteristic curves analysis, of 450 per mm3for polymorphonuclear cells and of 0.07 for arterial–ascitic fluid pH gradient, allow high positive and negative predictive values for spontaneous bacterial peritonitis. The polymorphonuclear cell count has the highest sensitivity and specificity. The use of both testscan be useful in the early diagnosis of spontaneous bacterial peritonitis or to confirmspontaneous bacterial peritonitis when bacteriological cultures are negative, if conditions associated with neutrocytic ascites or malignancy are excluded.
Pericanalicular Location of Hepatocyte Lysosomes and Effects of Fasting: A Morphometric Analysis
Authors: Sewell Richard B. , Dillon Catherine , Grinpukel Susan , Yeomans Neville D. , Smallwood Richard A.
Published: March 2007

Lysosomes constitute a small proportion of hepatocyte volume, but they play a key degradative role, particularly during catabolic states such as nutritional deprivation. Our preliminary observations in hepatocytes suggested that fasting may alter the distribution of lysosomes, which are thought to congregate about the biliary canaliculus. Thecurrent study aimed to provide, in both fed and fasted rats, a quantitative morphometric analysis of lysosomes in the immediate pericanalicular and successively more distant regions. We selected three regions of radius 0 to 2 (region 1), 2 to 4 (region 2) and 4to 6 μm (region 3) about the bile canaliculus and compared lysosomal numerical and volume densities. In fed rats, numerical and volume densities of both primary andsecondary lysosomes were 5 to 10 times greater in region 1 than in region 3. After a 72–hr fast, this gradient was lost due largely to a reduced number of primary and secondary lysosomes in region 1. Our calculations suggest that during fasting, lysosomes are redistributed away from the bile canaliculus towards the more distant cytoplasm rather than being lost from the cell, for example, into bile. The number of overt macroautophagic vacuoles was not altered by fasting. We estimate that, in fed rats, there areapproximately 270 lysosomes per hepatocyte, and that 43% of lysosomes are contained in the immediate pericanalicular region 1, which comprises only 7 to 8% of the cytoplasmic volume. The reason for this pericanalicular concentration of lysosomes and the mechanism of the redistribution with fasting remain to be determined.
From Australia to Chinchilla to Jamaica
Authors: Thaler Michael M.
Published: March 2007

A general impairment of liver mitochondrial enzymes is central to Reye's syndrome (RS). The respiration of isolated liver mitochondria was measured after the addition of concentrated normal serum or RS serum derived from 12 patients. RS serum stimulates oxygen consumption in isolated rat liver mitochondria. This effect is due to (a) the oxidation of uric acid by peroxisomes contaminating the preparation and (b) a stimulation of mitochondrial respiration (1.05 ± 0.14 nmole of O2/ min · mg of protein; control 0.30 ± 0.08 nmole O2/min · mg). The stimulation of respiration occurs in the presence of all respiratory substrates, is dependent on the amount of serum added, and represents an uncoupling of oxidativephosphorylation. RS serum reduces ATP formation by 15 to 76%. The uncoupling effect correlates with the amount of free fatty acid in the serum sample and resembles theeffect induced by the addition of a dicarboxylic fatty acid. Dicarboxylic fatty acids, especially long–chain dicarboxylic acids, impair ATP formation. Dicarboxylic acids were found in the serum of all RS patients and comprised as much as 54% of the total serum free fatty acids. Ninety percent of the serum dicarboxylic acids were of 16 to 18 carbon lengths. The amount of dicarboxylic acids in the RS serum corresponded directly with the reduction in ATP formation by the RS serum. This demonstrates that dicarboxylic acids occur in RS and may be important in the general impairment of mitochondrial function in RS and other disorders where they are present.
A dot-immunobinding assay for antimitochondrial antibodies
Authors: Penner Edward , Goldenberg Hans , Meryn Siegfried , Gordon Julian
Published: March 2007

A dot-immunobinding assay was established for the detection of antimitochondrial antibodies. Nitrocellulose strips were coated with sonicated rat liver mitochondria and incubated in the presence of human sera. The resulting immune complexes were visualized with an enzyme-linked second antibody. Antimitochondrial antibodies were found in the sera of 96% of patients with primary biliary cirrhosis, 17% of patients with autoimmune hepatitis and 4% of patients with progressive systemic sclerosis. Sera of patients with other liver diseases or with systemic lupus erythematosus, specimens positive for M1 and M3 mitochondrial antibodies and samples from normal controls were all negative. Antinuclear and other cytoplasmic antibodies were not detected in this assay. The dot-immunobinding assay for antimitochondrial antibodies is rapid and sensitive, and obviates the need for expensive equipment.
Serum hyaluronate in liver diseases: Study by enzymoimmunological assay
Authors: Frébourg Thierry , Delpech Bertrand , Bercoff Eric , Senant Jacques , Bertrand Philippe , Deugnier Yves , Bourreille Jacques
Published: March 2007

It has been suggested that glycosaminoglycans are involved in the pathogenesis of liver fibrosis. Furthermore, recent studies have reported that one of them, hyaluronate, was mainly taken up and degraded by the liver. Using an enzymoimmunological assay, based on hyaluronate-hyaluronectin interaction, serum levels of hyaluronate were measured in 113 patients with various liver diseases. Patients were divided into six groups according to clinical, biological and histological data: Group 1—alcoholic cirrhosis (n=47) including alcoholic cirrhosis with alcoholic hepatitis (n=24); Group 2—primary biliary cirrhosis (n=21); Group 3—cirrhosis related to viral hepatitis (n=10); Group 4—idiopathic hemochromatosis (n=17); Group 5—alcoholic fatty liver (n=8); and Group 6—viral or drug acute hepatitis (n=10). Ninety-four blood donors were studied as controls. Levels of hyaluronate were found to be strikingly elevated in Group 1 (1,225 ± 1,137 μg per liter), Group 2 (792 ± 739 μg per liter), Group 3 (649 ± 373 (μg per liter), and Group 4 (246 ± 242 μg per liter), whereas patients in Group 5 (94 ± 63 μg per liter) and Group 6 (73 ± 57 μg per liter) had values close to controls (23 ± 17 μg per liter). There was a significant correlation between serum hyaluronate and serum albumin, prothrombin time, factor V concentration and serum γ-globulins. It is suggested that hyaluronate levels reflect both active fibrosis and hepatic failure and may be a quantitative marker of severity of hepatic injury.
Controlled trial of vasopressin plus nitroglycerin vs. vasopressin alone in the treatment of bleeding esophageal varices
Authors: Tsai Yang-Te , Lay Chii-Shyan , Lai Kwok-Hung , Ng Wai-Wah , Yeh Yeong-Shyan , Wang Jiin-Yu , Chiang Teun-Tzong , Lee Shou-Dong , Chiang Benjamin N. , Lo Kwang-Juei
Published: March 2007

Thirty-nine patients admitted during a 16-month period for acute bleeding from varices confirmed by emergency endoscopy were randomized to receive either continuous intravenous infusions of vasopressin alone (0.66 units per min) (Group I: 19 patients) or vasopressin plus sublingual nitroglycerin (0.6 mg every 30 min for 6 hr) (Group II: 20 patients). The two groups of patients were similar in the type and severity of their cirrhosis. Bleeding was controlled initially in 47% (9/19) of the patients in Group I and 55% (11/20) of the patients in Group II after 6 hr of infusion (not statistically significant). Complete control of bleeding during 24 hr of infusion was achieved in only 4 of 19 patients in Group I (21%) but in 9 of 20 in Group II (45%). This difference is not statistically significant. The total number of patients with complications during infusions were significantly different statistically in the vasopressin and vasopressin-nitroglycerin groups, respectively (17/19 vs. 7/20, p<0.001). Major complications requiring immediate cessation of infusions were observed in 6 of 19 of the patients in Group I (32%) and in 2 of 20 in Group II (10%) (p<0.05). Mortality (58% in Group I, 55% in Group II) and transfusion requirements were similar in the two groups. This study shows that the addition of sublingual nitroglycerin to intravenous vasopressin does not alter the efficacy of vasopressin alone in controlling hemorrhage from esophageal varices, but it does significantly reduce the complications.
The hemodynamic effect of verapamil on portal hypertension in patients with postnecrotic cirrhosis
Authors: Kong Chi-Woon , Lay Chii-Shyan , Tsai Yang-Te , Yeh Ching-Lan , Lai Kwok-Hung , Lee Shou-Dong , Lo Kwang-Juei , Chiang Benjamin N.
Published: March 2007

Wedged hepatic venous pressure, free hepatic venous pressure and cardiac index were measured before and 1 hr after i.v. administration as well as 1 month and 3 months after chronic oral administration of verapamil in 10 patients with HBsAg-positive cirrhosis.

The gradient between wedged hepatic venous pressure and free hepatic venous pressure was decreased 14% at 1 hr after i.v. administration of 10 mg verapamil, and the sustained decrease in hepatic venous pressure gradient was also demonstrated in the patients after 1 month, and in six patients after 3 months of continuous oral administration of verapamil. The mean arterial pressure significantly decreased (p<0.01) 1 hr after the administration of verapamil. There was no significant change in other hemodynamic values. We conclude that chronic oral administration of verapamil can reduce the hepatic venous pressure gradient in patients with compensated HBsAg-positive cirrhosis and portal hypertension.
Androgen and estrogen receptors in hepatocellular carcinoma and in the surrounding noncancerous liver tissue
Authors: Ohnishi Shin , Murakami Tohru , Moriyama Takashi , Mitamura Keiji , Imawari Michio
Published: March 2007

Both androgen and estrogen receptors were studied in human hepatocellular carcinoma and noncancerous liver tissue surrounding it. Androgen receptor was detected in the cytosol and/or nucleosol of 4 of 8 cancerous tissues and 1 of 6 noncancerous tissues. The levels of androgen receptor in hepatocellular carcinomas ranged from 3.4 to 37.6 fmoles per mg protein with dissociation constants (Kd) of 0.226 − 51.3×10−9
M. That in the surrounding noncancerous tissue was 2.1 fmoles per mg protein with Kd of 0.941×10−9
M. Estrogen receptor was detected in the cytosol of 1 of 7 cancerous tissues, while it was detected in the cytosol and/or nucleosol of 3 of 7 noncancerous tissues. The level of estrogen receptor in hepatocellular carcinoma was 4.9 fmoles per mg protein with Kd of 1.20×10−9
M, and those in the surrounding noncancerous tissues ranged from 2.6 to 1,073 fmoles per mg protein with Kd of 0.223 − 3.15×10−9
M. The results suggest that the expression of androgen receptor may be augmented in association with malignant transformation of hepatocytes while the expression of estrogen receptor may be rather suppressed and that some of hepatocellular carcinomas may be androgen-dependent.
Hepatic transport of fluorescent molecules: In Vivo studies using intravital TV microscopy
Authors: Sherman I. A. , Fisher M. M.
Published: March 2007

Experiments were undertaken to evaluate the role of intravital epifluorescent TV microscopy in the study of the hepatic transport of sodium fluorescein and fluorescently labeled sodium glycocholate in rats and hamsters.

It was found that the apparent sinusoid to canaliculus transport time for fluorescently labeled sodium glycocholate was only half of that for sodium fluorescein, 0.50 and 0.92 sec, respectively. The sinusoid to bile transport time was 35 sec for fluorescently labeled sodium glycocholate and 90 sec for sodium fluorescein. There were also marked differences in zonal clearance of these two substances. Sodium fluorescein was removed from Zone 1 much faster than from Zone 3, while fluorescently labeled sodium glycocholate was cleared at the same rate by hepatocytes in all three acinar zones.

This study provides direct evidence that there are zonal differences in hepatic transport rates for certain compounds. It also demonstrates that epifluorescent TV microscopy has the potential to provide quantitative data on the transport rates of bile acids and other molecules into, through and out of the liver.
Sublobular distribution of transferases and hydrolases associated with glucuronide, sulfate and glutathione conjugation in human liver
Authors: El Mouelhi Mohamed , Kauffman Frederick C.
Published: March 2007

Activities of glucuronosyltransferase, sulfotransferase, glutathione S-transferase, β-glucuronidase and sulfatase were determined in microdissected samples of periportal and pericentral sublobular regions from four human livers obtained at immediate autopsy. New methods are presented for the microdetermination of sulfotransferase and sulfatase activities in microdissected samples weighing 0.1 to 4 μg dry weight using umbelliferone and 4-methylumbelliferone sulfate as substrates. The three transferases were distributed heterogenously across the liver lobule. Glucuronosyltransferase and glutathione S-transferase were localized predominantly in pericentral regions. In contrast, sulfotransferase activity was greater in periportal than pericentral regions. Average activities for glucuronosyltransferase and sulfotransferase were 23, and 50 μmoles. gm dry wt−1. hr−1, respectively, in periportal regions, and 34 and 38 μmoles. gm dry wt−1. hr−1, respectively, in pericentral regions. Activities of glutathione S-transferase were considerably higher than those of the other transferases and were 8.3 mmoles. gm dry wt−1. hr−1 in periportal areas and 12.2 mmoles. gm dry wt−1 hr−1 in pericentral areas. The two hydrolases studied, β-glucuronidase and sulfatase, were evenly distributed across the liver lobule. The presence of significant hydrolase and transferase activities in both zones of the liver lobule supports the idea that net production of both sulfate and glucuronide conjugates may be influenced by futile cycling of conjugation-deconjugation reactions in both zones of the liver. Based on enhanced formation of sulfate but not glucuronide conjugates in homogenates of human liver treated with inhibitors of the hydrolases, it is suggested that futile cycling is more pertinent to the regulation of sulfation than glucuronidation.
Composition of pigmented centers of cholesterol gallstones
Authors: Malet Peter F. , Williamson Clarke E. , Trotman Bruce W. , Soloway Roger D.
Published: March 2007

Most cholesterol gallstones have visually pigmented centers, but it is unclear whether this represents simple co-precipitation of pigment with cholesterol during stone nidation or nidation on a true pigment stone center. To clarify this issue, we selected from among 67 sets of cholesterol gallstones, 12 sets with the most conspicuously pigmented centers. The composition of the centers and the peripheries of these 12 stones was analyzed using infrared spectroscopy and compared with that of 10 black pigment gallstones. The pigmented centers of cholesterol stones contained 80.1 ± 7.9% (mean ± S.E.) cholesterol, 6.2 ± 3.4% calcium bilirubinate (only 4 of the 12 centers had measurable calcium bilirubinate), trace amounts of calcium phosphate and no calcium carbonate or calcium palmitate. The peripheral areas of the cholesterol stones contained 91.6 ± 2.3% cholesterol and no detectable calcium salts. For comparison, the composition of the centers of 10 black pigment gallstones was 13.5 ± 2.2% cholesterol, 28.2 ± 5.3% calcium bilirubinate, 5.5 ± 2.4% calcium phosphate and 10.6 ± 5.8% calcium carbonate. The composition of only one cholesterol stone center (15.8% cholesterol, 26.8% calcium bilirubinate) resembled that of a pigment stone, but even this center differed from that of a typical pigment stone in that it contained only a trace amount of calcium phosphate and no calcium carbonate. Thus, the chemical composition of pigmented centers of cholesterol gallstones is quantitatively different from that of black pigment stones, suggesting that cholesterol gallstones do not form on a pigment stone nidus.
Effect of combined treatment with 2,3,7,8-tetrachlorodibenzo- p -dioxin and phototherapy on bilirubin metabolism in the jaundiced gunn rat
Authors: Cohen Arnold N. , Kapitulnik Jaime , Ostrow Donald J. , Webster Cecile C.
Published: March 2007

2,3,7,8-Tetrachlorodibenzo-p-dioxin, a potent inducer of microsomal cytochrome P448-dependent monoxygenases, and phototherapy both accelerate bilirubin metabolism and decrease jaundice in Gunn rats. The effects of combined treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin and light were studied in these rats by applying phototherapy for 65 hr, beginning 5 days after induction with 2,3,7,8-tetrachlorodibenzo-p-dioxin.

2,3,7,8-Tetrachlorodibenzo-p-dioxin pretreatment caused a 75% decline in plasma bilirubin in 5 days, with no change thereafter, whether or not the rats were exposed subsequently to phototherapy. In the uninduced rats, plasma bilirubin levels declined by 55% after 40 hr of phototherapy. As determined by [14C]bilirubin kinetics, both 2,3,7,8-tetrachlorodibenzo-p-dioxin and phototherapy increased fractional bilirubin turnover and decreased the total bilirubin pool. In the 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced rats, the contracted bilirubin pool shifted from skin to liver, but these tissue pools did not change further during phototherapy. By contrast, in uninduced rats, phototherapy decreased the cutaneous bilirubin pool, which is the main target of phototherapy. 2,3,7,8-Tetrachlorodibenzo-p-dioxin was more effective than phototherapy in diminishing plasma bilirubin levels and the total bilirubin pool, but the combined treatment (2,3,7,8-tetrachlorodibenzo-p-dioxin followed by phototherapy) was no more effective than 2,3,7,8-tetrachlorodibenzo-p-dioxin alone.
Gastric bleeding in portal hypertension: Inflammatory or congestive?
Authors: Sarfeh James I. , Tarnawski A.
Published: March 2007

This paper reports the incidence and natural history of macroscopic gastritis in a series of 127 consecutive patients with portal hypertension of various aetiologies. Gastritis was observed endoscopically in 65 patients (51%) and was of two main types. Twenty eight patients had severe or persistent gastritis which caused clinically significant bleeding on 80 occasions and accounted for 25% of the bleeds from all sources. The remainder had mild gastritis. The presence of gastritis seemed to be independent of the severity of liver disease or the degree of rise of wedged hepatic venous pressure and there was no difference in age, sex, or drugs prescribed in patients with or without gastritis. The mean follow up period and the mean number of sclerotherapy treatments was significantly greater (p<0.005) in patients with gastritis. Full thickness gastric biopsies in seven surgical patients and 11 autopsy specimens showed dilated and tortuous submucosal veins. Endoscopic biopsies in 14 patients showed vascular ectasia in the mucosal layer which was in excess of the degree of inflammatory infiltrate. Gastritis occurred in patients with portal hypertension of all common aetiologies and the clinical and pathological evidence supports the contention that it reflects a congested gastric mucosa and should be renamed congestive gastropathy. As injection sclerotherapy improves survival from variceal bleeding congestive gastropathy may become more common. The response to conventional (‘anti-erosive’) therapy is poor and measures aimed at reducing the gastric portal pressure may be the only effective means of treating this condition.
Halothane hepatitis: A double defect?
Authors: Ranek Leo
Published: March 2007

We studied susceptibility to halothane hepatitis with an in vitro test that detects cell damage from electrophilic drug intermediates. Metabolites of phenytoin were generated by incubation of phenytoin with rat hepatic microsomes in the presence of the epoxide hydrolase inhibitor 1,1,1-trichloropropene oxide (TCPO), which prevents the further metabolism of phenytoin to an inert metabolite. In lymphocytes exposed to this system, cytotoxicity was measured by trypan blue dye exclusion and was expressed as the percentage increase in trypan blue-positive cells after the addition of TCPO. In the presence of TCPO, lymphocytes from 11 patients with halothane hepatitis exhibited an increase in cytotoxicity at 0.06 mM phenytoin that was eight times greater than the increase in healthy controls (54 ± 10 per cent [mean ± S.E.M.] vs. 7.1 ± 2.2 per cent, p<0.001). Patients with other liver diseases and persons recently exposed to halothane without adverse effects did not differ from healthy controls. In three patients with halothane hepatitis who were studied serially, the lymphocyte abnormality was still present after 13 months. Family studies revealed abnormal results on 10 cytotoxicity tests among 19 members of four families.

We propose that there is a familial, constitutional susceptibility factor that predisposes persons to halothane hepatitis.
The calcium ionophore A23187 stimulates glycoprotein secretion by the guinea pig gallbladder
Authors: Malet Peter F. , Locke Catherine L. , Trotman Bruce W. , Soloway Roger D.
Published: March 2007

The purpose of this study was to examine the role of calcium ions in gallbladder glycoprotein secretion in cultured guinea pig gallbladder explants. The calcium ionophore A23187 showed a threshold of 2 μg per ml medium for stimulation of secretion of [3H]glucosamine-labeled glycoproteins over a 30 min incubation period. The ionophore at 3 and 5 μg per ml medium resulted in a 3- to 4-fold increase in secretion of [3H]glucosamine-labeled glycoproteins. Ionophore-induced stimulation of glycoprotein secretion was abolished by the addition of 0.01 mM verapamil to the medium. To study the effect of changes in extracellular calcium on basal glycoprotein secretion, explants were cultured for 24 hr in media with 0.007, 0.5 or 2.0 mM calcium; no differences in basal glycoprotein secretion were observed. When cultured in medium with 1.0 mM EGTA, basal secretion decreased significantly vs. controls in 0.007 mM total calcium medium. Total [3H]glucosamine incorporation by explants in medium with EGTA was unaltered, however, suggesting that the low level of calcium in the medium was selectively impairing the secretory process. These findings indicate that calcium ions are important in the regulation of gallbladder glycoprotein secretion.
Rat hepatic bile acid sulfotransferase: Identification of the catalytic polypeptide and evidence for polymeric forms in female rats
Authors: Collins Robert H. , Lack Leon , Harman Kenneth M. , Killenberg Paul G.
Published: March 2007

A monoclonal antibody, PK1B, directed against rat liver bile acid sulfotransferase was used for the purification and characterization of the enzyme. Incubation of rat liver supernatant with the antibody followed by immunoprecipitation with Staphylococcus aureus cells demonstrated that PK1B reacted with 90% of the enzymatic activity present in the liver supernatant from female rats and 40 to 50% of the activity in male liver preparations. Immunoadsorption chromatography with PK1B bound to Sepharose isolated active enzyme which was purified greater than 75-fold. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of this preparation in the presence of 2-mercaptoethanol demonstrated three polypeptides: Mr 29,500; 32,500, and 34,000. Western blot analysis indicated that PK1B recognized an epitope which was found only on the Mr 29,500 polypeptide. Two-dimensional gel electrophoresis associated the enzymatic activity with this Mr 29,500 band. High-pressure liquid chromatographic analysis of immunopurified enzyme defined three distinct, enzymatically active protein populations: I (Mr 400,000 to 170,000); II (Mr 130,000), and III (Mr 43,000). An Mr 29,500 polypeptide was the sole constituent of Peaks I and III and a major constituent of Peak II. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence and absence of 2-mercaptoethanol indicated that in Peak II, catalytically active Mr 29,500 protein is associated with the other two polypeptides by disulfide bonds. In contrast, Peak I consists of a polymer of Mr 29,500 polypeptide which is independent of disulfide interaction.
Depletion of liver and esophageal epithelium vitamin a after chronic moderate ethanol consumption in rats: Inverse relation to zinc nutriture
Authors: Mobarhan Sohrab , Layden Thomas J. , Friedman Howard , Kunigk Annette , Donahue Philip
Published: March 2007

This study was designed to determine whether chronic moderate ethanol ingestion alters the levels of vitamin A of liver and esophageal epithelium and if this is dependent on zinc nutriture. Forty male Sprague-Dawley 4-week-old rats were divided into five groups: zinc-deficient (0.9 ppm), ethanol-fed; zinc-deficient; zinc-adequate (25 ppm); zinc-adequate (25 ppm), ethanol-fed; and zinc-supplemented (50 ppm), ethanol-fed. All rats received liquid Lieber-DeCarli diet containing 4,000 IU per liter of vitamin A for 5 weeks. Zinc-deficient, ethanol-fed rats and zinc-adequate, ethanol-fed rats and zinc-supplemented, ethanol-fed rats received 15.5% of the caloric intake as ethanol while zinc-deficient and zinc-adequate rats received isocaloric amounts of maltose dextrin. All groups were pair-fed to zinc-deficient, ethanol-fed rats. In addition, a group of eight rats designated as weight-restricted controls were fed a diet similar to the one given to zinc-adequate rats but in the amount to obtain a final weight as in the zinc-deficient group. After 35 days, the liver histology was normal in all rats, and no fat accumulation was noted. Hepatic vitamin A concentration was significantly decreased in zinc-adequate, ethanol-fed rats (41 ± 10 μg per gm) and further in zinc-supplemented, ethanol-fed rats (12 ± 5 μg per gm) as compared to controls (137 ± 49). A highly significant negative correlation between serum zinc and liver vitamin A was found in ethanol-fed animals. In zinc-deficient, ethanol-fed rats, ethanol did not induce mobilization of liver vitamin A, however, it caused a significant reduction in the ratio of retinyl ester to retinol (53 ± 16%) as compared to controls. Zinc deficiency in the absence of alcohol caused a reduction of hepatic vitamin A concentration and total content, and this effect was not secondary to protein-caloric malnutrition as values were unaffected in weight-restricted controls. In esophageal epithelium, a significant reduction of vitamin A stores was noted in zinc-supplemented, ethanol-fed rats (0.67 ± 0.7 μg per gm) as compared to controls (3.3 ± 1.0, p < 0.01), and a significant negative correlation was seen between serum zinc and esophageal vitamin A levels of ethanol-fed rats. These results demonstrate that ethanol in moderate quantities causes profound alterations of the vitamin A status of the liver and esophagus, and these effects are inversely correlated to zinc nutriture.
Comparison between polyclonal and first and second generation monoclonal radioimmunoassays in the detection of hepatitis B surface antigen in patients with hepatocellular carcinoma
Authors: Kew Michael C. , Fujita Yumiko , Takahashi Hiroshi , Coppins Ann , Wands Jack R.
Published: March 2007

Serum from 221 black patients with hepatocellular carcinoma was tested for HBsAg using a polyclonal radioimmunoassay, and first and second generation monoclonal radioimmunoassays designated M1-RIA and M2-RIA. These monoclonal radioimmunoassays have a lower limit of detection of about 55 and 15 pg of HBsAg-associated epitopes per ml of serum. Correlations were made with other hepatitis B virus serologic markers such as anti-HBc and anti-HBs, using polyclonal radioimmunoassays. We found 47.5% (105/221) of the patients were HBsAg positive by conventional polyclonal radioimmunoassay; all of these patients were also reactive by monoclonal radioimmunoassay. Of the 116 polyclonal radioimmunoassay-negative patients, 4 (3.4%) were reactive by M1-RIA. These four patients were all positive for anti-HBc and/or anti-HBs antibodies. There were 106 of 112 patients negative for HBsAg by polyclonal radioimmunoassay and M1-RIA available for testing by the M2-RIA; 11 (10.4%) were found to be positive only by this test. Thus, with the use of M1- and M2-RIAs, the number of hepatocellular carcinoma patients negative for HBsAg by polyclonal radioimmunoassay was reduced by 14% in this population. More importantly, of the 11 M2-RIA-positive patients, six demonstrated anti-HBc and/or anti-HBs antibodies whereas the remaining five had no serologic evidence of recent or past hepatitis B virus exposure. Finally, of the 21 patients in this series with no markers of hepatitis B virus infection using polyclonal radioimmunoassay and M1-RIA, five (24%) were reactive for HBsAg-associated epitopes by M2-RIA. Taken together, these findings may be due in part to the detection of HBsAg in immune complexes, particularly in those patients with anti-HBc and anti-HBs. However, in hepatocellular carcinoma patients with no hepatitis B virus serologic markers, the M2-RIA may have been reactive due to low levels of antigen expression co-existing with an inadequate or lack of a host immune response to HBcAg and HBsAg, variable expression of hepatitis B virus gene products or to a “hepatitis B-related virus”.
Natural course of delta superinfection in chronic hepatitis B virus-infected patients: Histopathologic study with multiple liver biopsies
Authors: Govindarajan Sugantha , de Cock Kevin M. , Redeker Allan G.
Published: March 2007

We evaluated the natural course of delta superinfection in patients with chronic B viral hepatitis by examining serial liver biopsy specimens from 23 patients. The mean follow-up period was 2.5 years, and the average number of liver biopsies per patient was 2.8. The striking features were: (i) the prolonged acute hepatitis-like necroinflammatory activity (mean = 15 months) following acute delta hepatitis and (ii) rapid progression to severe forms of liver disease within a mean interval of 24 months from the onset of delta superinfection. Although there was a high frequency of stainable delta antigen (90%) in these liver biopsy specimens, there was no correlation between the degree of positivity and the histopathological changes.