bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Putrescine on D–Galactosamine-Induced Acute Liver Failure in Rats
Authors: Nishiguchi Shuhei , Kuroki Tetsuo , Takeda Tadashi , Nakajima Shinya , Shiomi Susumu , Seki Shuichi , Matsui-Yuasa Isao , Otani Shuzo , Kobayashi Kenzo
Published: March 2007

We studied the effects of putrescine on acute liver failure caused in rats by two injections of 1 gm/kg D–galactosamine. The hepatic polyamine level rose only slightly in the D–galactosamine-injected rats treated with glucagon and insulin, and [3H]thymidine incorporation into DNA increased little; these hormones did not improve the survival rate. When D–galactosamine-injected rats were given putrescine, the putrescine concentration in the liver increased and the survival rate of the rats was significantly higher than that of control rats given only D–galactosamine. Putrescine administration tended to lower the serum level of alanine aminotransferase in rats injected with D–galactosamine, so the polyamine might have a protective effect on hepatocytes. Putrescine significantly increased [3H]thymidine incorporation in the liver; thus it accelerated liver regeneration. Difluoromethylornithine decreased the level of putrescine in the liver, decreasing both [3H]thymidine uptake and the survival rate. In the rats treated with D–galactosamine, in which liver damage was so severe that treatment with glucagon and insulin was ineffective, the intraperitoneal administration of putrescine increased the survival rate in acute liver failure. This probably resulted mainly from activation of liver regeneration and possibly from a protective effect of putrescine on the liver. (Hepatology 1990;12:348-353).
The Potentiation of Yersinial Infections in Iron Storage Disorders
Authors: Hart Hamish H.
Published: March 2007

Liver abscesses due to Yersinia enterocolitica usually occur in patients with a pathological iron overload. A case of multiple liver abscesses in a patient with haemochromatosis is reported and the literature is reviewed.
Partial Portal Decompression: Two Approaches to the Management of Portal Hypertension
Authors: Rypins Eric B. , Sarfeh James I.
Published: March 2007

To test the hypothesis that partial portal decompression in the treatment of variceal hemorrhage will diminish subsequent encephalopathy, 50 consecutive patients were studied after construction of a small–stoma (10 to 12 mm) side–to–side portacaval shunt, with the goal of a postoperative portacaval pressure gradient of 10 mm Hg. During follow–up averaging 26 months, six patients (12 percent) died. Four patients (8 percent) had episodes of rebleeding, only one from varices. All patients had patent shunts at subsequent angiography or ultrasonography. Despite consistent (100 percent) postoperative reversal or stagnation of portal flow on duplex scan, encephalopathy on clinical and psychometric grounds was observed in only three patients (6 percent). This study suggests that small–stoma portacaval shunt can be performed with reliably low rates of rebleeding and encephalopathy. That encephalopathy was rare despite loss of hepatic portal perfusion incriminates other factors besides portal flow in the genesis of postshunt hepatic failure.
Both macrophages and endothelial cells of the human hepatic sinusoid express the CD4 molecule, a receptor for the human immunodeficiency virus
Authors: Scoazec Jean-Yves , Feldmann Gérard
Published: March 2007

Hepatic sinusoidal lesions, including peliosis hepatis and sinusoidal dilatation, are frequently observed during human immunodeficiency virus infection. It has been proposed that human immunodeficiency virus itself plays a role in their pathogenesis. To test this hypothesis, we attempted to determine whether liver sinusoidal cells express the CD4 molecule, which behaves as a membrane receptor mediating the binding of human immunodeficiency virus to its target cells. For this purpose, three monoclonal antibodies-OKT4, OKT4a and anti-Leu3a + 3b, binding to different epitopes of the CD4 molecule-were used. All antibodies tested had the same tissue reactivity. On light microscopy, they reacted with most sinusoidal macrophages and in addition gave a continuous labeling of the sinusoidal lining suggestive of endothelial cell reactivity. On ultrastructural examination, the plasma membranes of both sinusoidal macrophages and endothelial cells were labeled. The reactive antigen was characterized by immunoblotting of liver homogenates. A unique band was detected, corresponding to an antigen with an apparent molecular weight of 54,000 Da, comparable to that reported for the CD4 molecule expressed on lymphocytes and monocytes. Therefore combination of structural and immunochemical data makes it possible to assess that both endothelial cells and macrophages of the hepatic sinusoid express the CD4 molecule. Consequently, both cell types constitute putative targets for human immunodeficiency virus and/or human immunodeficiency virus-related lesions. They may be involved in the pathogenesis of liver sinusoidal lesions observed in human immunodeficiency virus infection and may constitute an unsuspected reservoir of the virus. (Hepatology 1990;12:505-510).
Plexiform neurofibromatosis of the liver and mesentery in a child
Authors: Partin Jacqueline S. , Lane Bernard P. , Partin John C. , Edelstein Lawrence R. , Priebe Cedric J.
Published: March 2007

Plexiform neurofibromatosis of the liver was recognized by needle biopsy of the liver in an 11-yr-old boy who had a 2-yr history of diarrhea, intermittent abdominal pain, failure to gain weight and progressive abdominal distention. Imaging studies demonstrated a large retroperitoneal mass; a laparotomy was performed. At surgery, the mesentery was greatly thickened by neurofibromas, and plexiform neurofibroma extended through the hilum of the liver. Light and electron microscopy demonstrated that in addition to the direct involvement by tumor, neural hyperplasia existed throughout the liver. The most distal ramifications of the portal spaces were filled with Schwann cells, bundles of unmyelinated nerves and perineurium-surrounded nerves containing myelinated and unmyelinated fibers. The ultrastructural findings were consistent with stimulation of proliferation of all the portal neural elements and tumoral tissue. The nontumoral response was more than simple hyperplasia because it appeared to result in fibrotic changes in the most involved areas and active breaching of the limiting plate with destruction of hepatocytes and collagen deposition throughout the liver. (Hepatology 1990;12:559-564).
Identification and characterization of intrahepatic hepatitis b virus dna in hbsag-seronegative patients with chronic liver disease and hepatocellular carcinoma in Taiwan
Authors: Lai Ming-Yang , Chen Pei-Jer , Yang Pei-Ming , Sheu Jin-Chuan , Sung Juei-Low , Chen Ding-Shinn
Published: March 2007

To clarify the role of hepatitis B virus infection in HBsAg-seronegative patients with chronic liver disease and hepatocellular carcinoma in Taiwan, we examined the hepatitis B virus DNA in liver biopsy tissues of 112 patients by Southern blot analysis. The patients studied included 43 patients with nonalcoholic chronic liver disease, 21 patients with hepatocellular carcinoma and 48 control patients with other hepatobiiary and gastrointestinal diseases. To confirm the specificity of the intrahepatic hepatitis B virus DNA signal and to understand the structure of the integrated viral sequences, molecular cloning and DNA sequencing of an integrated hepatitis B virus DNA were done in one patient. Among 13 patients without serological evidence of previous hepatitis B virus infection, no hepatitis B virus sequences were found in the liver. In other HBsAg-negative patients with evidence of previous hepatitis B virus exposure, a substantial positive rate of intrahepatic hepatitis B virus DNA was found (7%). The intrahepatic hepatitis B virus DNA was all in integrated form. The positive rate among patients with nonalcoholic chronic hepatitis and cirrhosis (2%) was not different from that of the control group with other hepatobiliary and gastrointestinal diseases (4%). However, the positive rate of integrated hepatitis B virus DNA between hepatocellular carcinoma patients and nonhepatocellular carcinoma patients was statistically significant (19% vs. 3%, p <0.05). Molecular cloning and sequencing of a 3.0 kb EcoRI fragment of an integrated hepatitis B virus DNA from an anti-HBspositive patient revealed that it was a partial copy of the hepatitis B virus genome. Extensive rearrangement of the integrated viral sequences was found and might account for nonproduction of HIBsAg. We conclude that a substantial (7.8%) proportion of HBsAg-negative patients with chronic liver disease and hepatocellular carcinoma in Taiwan have hepatitis B virus DNA sequence in the liver, and integration of hepatitis B virus DNA may play a role in the development of hepatocellular carcinoma. (Hepatology 1990;12:575-581).
Hepatitis B virus replication in chinese patients with hepatocellular carcinoma
Authors: Lok Anna S. F. , Ma Oliver C. K.
Published: March 2007

We studied the frequency of hepatitis B virus replication in Chinese patients with hepatocellular carcinoma. Hepatitis B e antigen and hepatitis B virus DNA could be detected in the sera of 28% and 47% of 116 HBsAg-positive patients, but not in the sera of 15 HBsAg-negative patients. Replicative forms of hepatitis B virus DNA were detected in the neoplastic and nonneoplastic liver tissues from 34% and 62% of 29 HBsAg-positive patients and 0% and 20% of five HBsAg-negative patients by Southern blot hybridization analysis.

Of the 10 patients with chronic hepatitis B virus infection in whom hepatocellular carcinoma developed during follow-up, hepatitis B e antigen and hepatitis B virus DNA were detected in the sera of seven and eight patients, respectively, at presentation, 13 to 43 mo before the diagnosis of hepatocellular carcinoma. In nine patients, hepatitis B virus DNA was persistently or intermittently detected in the serum during follow-up. Five patients remained hepatitis B e antigen-positive and seven were detectable for hepatitis B virus DNA in serum when hepatocellular carcinoma was diagnosed. Four patients had one or more episodes of exacerbations before the diagnosis of hepatocellular carcinoma; in three, the exacerbations were associated with changes in level of hepatitis B virus replication.

Our study demonstrated that despite the long interval between the onset of hepatitis B virus infection and the development of hepatocellular carcinoma, hepatitis B virus replication persisted in most patients with hepatocellular carcinoma, albeit at a low level. (Hepatology 1990;12:582-588).
Histological features and clinical course of large regenerative nodules: Evaluation of their precancerous potentiality
Authors: Kondo Fukuo , Ebara Masaaki , Sugiura Nobuyuki , Wada Katsunori , Kita Kazuhiko , Hirooka Noboru , Nagato Yoshinobu , Kondo Yoichiro , Ohto Masao , Okuda Kunio
Published: March 2007

Seventeen patients with cirrhosis and histologically defined, ultrasonically detected, large regenerative nodules of the liver were followed without treatment for more than 1 yr (range = 13 to 51 mo). During the follow-up period, none of the nodules were demonstrated sonographically to have enlarged. Computed tomography and/or hepatic angiography were also performed in 14 patients, but there was no evidence of malignant transformation in these lesions. None of the 17 patients showed acute elevation of serum α-fetoprotein levels. Furthermore, four nodules became undetectable by these imaging procedures. Follow-up biopsy performed in seven patients revealed no significant histological changes from the first biopsy sample. However, HCCs developed apart from these large regenerative nodules in four patients. These HCCs suddenly appeared and did not seem to have been derived from preexisting large regenerative nodules.

From these observations, it was not possible to regard a large regenerative nodule having no cytological atypia as a high-potential premalignant lesion. Their precancerous potentiality needs to be compared with that of ordinary pseudolobules. (Hepatology 1990;12:592-598).
Corticosteroid therapy of alcoholic hepatitis: How many studies will it take?
Authors: Reynolds Telfer B.
Published: March 2007

Study Objective:

To determine the efficacy of a corticosteroid in reducing the short-term mortality of patients with 8evere alcoholic hepatitis.

Design:

Randomized, double-blind, placebo-controlled multicenter trial.

Setting:

Four university teaching hospitals.

Patients:

We enrolled 66 patients with alcoholic hepatitis and either spontaneous hepatic encephalopathy or a discriminant function value greater than 32, calculated using the formula: 4.6(prothrombin time -control time) + serum bilirubin [in pmoUL]/17.1. Fifty-nine patients (89%)completed the study. Two patients withdrew from the trial. The other 64 patients were hospitalized for the duration of the trial: however, treatment was discontinued in 5 patients because of potential drug toxicity.

Interventions:

Patients were randomly assigned to receive either methylprednisolone (32 mg) or placebo within 7 days of admission. Treatment was given for 28 days. The doses were then tapered over 2 weeks and discontinued.

Measurements and Main Results:

The endpoint of the study was death. Of the 31 recipients of placebo, 11 (35%)died within 28 days of randomization compared with 2 (6%)of the 36 patients given methylprednisolone (p = 0.006). The 95% CI for the difference in mortality was 12% to 70%. In the patients with spontaneous hepatic encephalopathy at entry, 9 of 19 recipients of placebo died (47%)compared with 1 (7%)of the 14 patients given methylprednisolone (p = 0.02). The 95% CI for the difference in mortality was 14% to 66%. The Cox proportional hazards regression model showed the advantage of methylprednisolone over placebo after adjustment for other potentidy important prognostic variables (p = 0.004).

Conclusions:

Methylprednisolone therapy decreases short-term mortality in patients with severe alcoholic hepatitis manifested either by spontaneous hepatic encephalopathy or a markedly elevated discriminant function value.
Supplemental putrescine reverses ethanol-associated inhibition of liver regeneration
Authors: Diehl Anna Mae , Abdo Suzan , Brown Nesbitt
Published: March 2007

Biosynthesis of the polyamines, putrescine, spermidine and spermine, is required for DNA synthesis and liver regeneration after partial hepatectomy. Chronic ethanol consumption impairs polyamine synthesis during the prereplicative phase after partial hepatectomy. To determine whether this delay in polyamine synthesis contributes to ethanol's inhibition of liver regeneration, the ability of supplemental putrescine to improve regeneration in ethanol-fed rats was tested. Chronically ethanol-fed rats and isocalorically maintained controls underwent partial hepatectomy and were injected intraperitoneally with saline or putrescine (0.03 or 0.30 mmol/kg) at 0, 4, 8 and 12 hr after partial hepatectomy. Rats were killed at 24, 48 or 72 hr, 1 hr after exposure to [3H]thymidine, so that DNA synthesis could be estimated. DNA synthesis was significantly inhibited in ethanol-fed rats treated with saline compared with saline-treated pair-fed controls. Supplemental putrescine did not affect DNA synthesis in pair-fed rats. In contrast, putrescine significantly improved [3H]thymidine incorporation 24 to 72 hr after partial hepatectomy in ethanol-fed rats. Intraperitoneal injection of putrescine (1.2 mmol/kg) at the time of partial hepatectomy increased hepatic polyamine concentrations for the first 6 hr after partial hepatectomy despite significantly inhibiting the activity of ornithine decarboxylase, the rate-limiting enzyme for polyamine synthesis, in both groups. Hepatic polyamine levels after putrescine injection were greater in ethanol-fed rats than in similarly treated controls. These data suggest that putrescine treatment triggers events that normalize DNA synthesis in ethanol-fed rats. These results confirm the hypothesis that ethanol's antiregenerative mechanism intimately involves inhibition of putrescine synthesis. (HEPATOLOGY 1990;12:633-637).
Growth of group A rotaviruses in a human liver cell line
Authors: Schwarz Kathleen B. , Moore Tara J. , Willoughby Rodney E. , Wee Siok-Bi , Vonderfecht Steven L. , Yolken Robert H.
Published: March 2007

Recent observations in children with rotavirus gastroenteritis and in infant mice given rotavirus vaccine by oral administration suggest that this well-known gastrointestinal pathogen may infect the liver. To examine this possibility, the susceptibility of Hep G2 cells to infection with a variety of rotavirus strains was tested. These cells were used because they are considered to be well differentiated and exhibit many liver-specific functions. The Hep G2 cells supported the growth of the simian strain rhesus rotavirus (MMU 18006), a strain currently being used in vaccine trials, but did not support the growth of any human strain (D, DS1, Price or ST3). The rhesus rotavirus infection was cytopathic and resulted in release of lactate dehydrogenase. Rhesus rotavirus growth in Hep G2 cells displayed trypsin-enhanced infectivity and was inhibited by pretreatment of cells with Arthrobacter ureafaciens neuraminidase but not with neuraminidase from Clostridium perfringens. Hep G2 cells were also permissive for another simian strain (SA11), a bovine strain (UK) and single gene substitution reassortants containing VP7 (the major outer capsid neutralization protein) from a human rotavirus strain and the remaining 10 genes from either rhesus rotavirus or UK. In general, UK and its reassortants produced lower levels of antigen than did rhesus rotavirus and its reassortants. Hep G2 cells and other hepatic cell lines may prove to be useful tools to explore the hepatotropic potential of wild-type rotaviruses and candidate vaccine strains. (HEPATOLOGY 1990;12:638-643).
HBV-DNA detection by gene amplification in acute hepatitis B
Authors: Quint Wim G. V. , de Bruijn Inge , Kruining Hans , Heijtink Rudolf A.
Published: March 2007

Serum samples from 62 women, inadvertently infected with hepatitis B virus in an in vitro fertilization program, were tested for the presence of hepatitis B virus-DNA using the polymerase chain reaction. Under conditions of a strict spatial separation of DNA extraction, amplification and product analysis, we succeeded in detection of as few as 360 hepatitis B virus particles per milliliter. Hepatitis B virus-DNA was detected with a high frequency during HBsAg and HBeAg antigenemia (98.5%) but also in the convalescent phase after appearance of antibody to HBsAg (18.2%). However, all patients with hepatitis B virus-DNA in convalescent sera were hepatitis B virus-DNA negative 3 to 6 mo later. All patients with HBeAg-positive samples showed hepatitis B virus-DNA positivity by polymerase chain reaction. For acute hepatitis, gene amplification restores the relationship between HBeAg and hepatitis B virus-DNA observed in serum from chronic hepatitis B patients and calls attention to the prolonged presence of hepatitis B virus-DNA in serum after generally accepted criteria for resolution of the infection have been reached. (HEPATOLOGY 1990;12:653-656).
Prospective study of early detection of hepatocellular carcinoma in patients with cirrhosis
Authors: Oka Hiroko , Kurioka Narito , Kim Kosyun , Kanno Toru , Kuroki Tetsuo , Mizoguchi Yasuhiro , Kobayashi Kenzo
Published: March 2007

We prospectively monitored 140 cirrhotic patients for the development of hepatocellular carcinoma for 6 yr, using periodical screening by high-resolution convex-array ultrasonography and α-fetoprotein. Twenty-eight patients were positive for HBs antigen, 26 patients had received blood transfusions and were negative for HBs antigen and 26 patients had a history of heavy drinking. We detected hepatocellular carcinoma in 40 patients during this period. The overall cumulative incidence of hepatocellular carcinoma in the 6 yr was 39%; the cumulative incidence was 59% in patients with HBsAg, 53% in patients who had had blood transfusions and were negative for HBsAg and 22% in patients who had a history of heavy drinking and who were without HBsAg. Detection of the carcinoma in 85% of these 40 patients was based on results of ultrasonography. Twenty-six of the patients (65%) had a small hepatocellular carcinoma of 2 cm or less. α-Fetoprotein levels were lower than 100 ng/ml in 56% of these 40 patients. Patients with cirrhosis are at high risk of developing hepatocellular carcinoma, especially patients with HBsAg or with a history of blood transfusion who are negative for HBsAg. Periodic monitoring by use of ultrasonography in particular is recommended for early detection of hepatocellular carcinoma. (HEPATOLOGY 1990;12:680-687).
Hepatocytes explanted in the spleen preferentially express carbamoylphosphate synthetase rather than glutamine synthetase
Authors: Lamers Wouter H. , Been Willem , Charles Robert , Moorman Antoon F. M.
Published: March 2007

Urea cycle enzymes and glutamine synthetase are essential for NH3 detoxification and systemic pH homeostasis in mammals. Carbamoylphosphate synthetase, the first and flux-determining enzyme of the cycle, is found only in a large periportal compartment, and glutamine synthetase is found only in a small, complementary pericentral compartment. Because it is not possible to manipulate experimentally the intrahepatic distribution of carbamoylphosphate synthetase and glutamine synthetase, we looked for conditions in which explanted hepatocytes would exhibit either the carbamoylphosphate synthetase phenotype or glutamine synthetase phenotype. In the spleen hepatocytes either settle as individual cells or in small agglomerates. The dispersed cells only express the carbamoylphosphate synthetase phenotype. Within the agglomerates, sinusoids that drain on venules develop. Hepatocytes surrounding the venules stain only weakly for carbamoylphosphate synthetase but are strongly positive for glutamine synthetase. These observations were made for explanted embryonic hepatocytes (no prior expression of either carbamoylphosphate synthetase or glutamine synthetase), neonatal hepatocytes (compartments of gene expression not yet established) and adult periportal and pericentral hepatocytes. (HEPATOLOGY 1990;12:701-709).
Monomicrobial nonneutrocytic bacterascites: A variant of spontaneous bacterial peritonitis
Authors: Runyon Bruce A.
Published: March 2007

Spontaneous bacterial peritonitis is diagnosed when (a) the ascitic fluid culture is positive, (b) the ascitic fluid neutrophil count is ≥ 250 cells/mm3 and (c) there is no evident intraabdominal surgically treatable source for infection. Few details are available regarding the variant of ascitic fluid infection in which the culture grows bacteria (pure growth of a single type of organism), but the neutrophil count is <250 cells/mm3. In this prospective study of 138 episodes of culture-positive spontaneously infected ascites detected in 105 patients, 44 (31.9%) were episodes of “monomicrobial nonneutrocytic bacterascites” compared with 94 (68.1%) episodes of spontaneous bacterial peritonitis. Seventeen patients had both types of infection. The infection-related mortality and hospitalization mortality were similar between the two groups. Patients with bacterascites appeared to have less severe liver disease. In 62% of bacterascites episodes in which a second paracentesis was performed before any treatment the fluid spontaneously became sterile without development of ascitic fluid neutrocytosis. Thirty-eight percent of patients with bacterascites (who underwent a second paracentesis before treatment was started) progressed to spontaneous bacterial peritonitis—sometimes within a few hours. The concentration of the chemoattractant C5a was not decreased in the ascitic fluid of the bacterascites patients; this excludes ascitic fluid C5a deficiency as the explanation of the lack of neutrocytosis.

Monomicrobial nonneutrocytic bacterascites is a common variant of ascitic fluid infection that may resolve without treatment or may progress to spontaneous bacterial peritonitis. (HEPATOLOGY 1990;12:710-715).
Relative frequencies of portosystemic pathways and renal shunt formation through the “posterior” gastric vein: Portographic study in 460 patients
Authors: Kimura Kunio , Ohto Masao , Matsutani Shoichi , Furuse Junji , Hoshin Kazuhiko , Okuda Kunio
Published: March 2007

Percutaneous transhepatic portography was carried out in 460 patients with portal hypertension to study various collateral routes. Besides the left gastric vein, which was the most frequent collateral route and feeder of esophageal varices, a distinct vein located between the left gastric vein and the short gastric vein constituted a major collateral route in 191 patients (42%). In terms of frequency, this vein was more significant than the short gastric (34%) and the paraumbilical vein (24%) as a collateral route. We propose that this previously anonymous vein be called the “posterior gastric” vein because it runs posterior to the stomach. This vein also formed a renal shunt, a common cause of encephalopathy, in 43 (23%) of the 191 patients; the relative frequency of renal shunt formation by this vein was significantly greater than that by the left gastric vein (12%) and the short gastric vein (18%) (HEPATOLOGY 1990; 12:725-728).
Methotrexate hepatotoxicity and the premature reporting of Mark Twain's death: Both greatly exaggerated
Authors: Kaplan Marshall M.
Published: March 2007

Ten histological criteria were evaluated semiquantitatively in the liver biopsies of 60 patients with rheumatoid arthritis (RA) before initiation of methotrexate (MTX) and were compared with 40 biopsies taken during MTX treatment (mean cumulative dose 1,322 mg). Mesenchymal changes (Kupffer cell proliferation, portal tract infiltration) and parenchymal alterations (nuclear variability, ballooning, fatty infiltration) were very common without statistically significant difference between the 2 groups. Slight periportal and/or portal fibrosis was present in 25% of patients without statistical differences between groups. Central fibrosis occurred in 13.5-12.5%. We conclude that liver abnormalities in RA are not related to MTX treatment.
Immunological studies before and during interferon therapy in chronic HBV infection: Identification of factors predicting response
Authors: Scully Linda J. , Brown David , Lloyd Chris , Shein Ruvan , Thomas Howard C.
Published: March 2007

Lymphoblastoid interferon is effective therapy in some but not all patients with chronic hepatitis B virus infection. To assess whether immunological parameters were predictive of response to interferon therapy, we determined the human leukocyte antigen type, CD4/CD8 ratio, natural killer cell activity, IgM anti-HBc antibody levels and concanavalin A—induced lymphocyte proliferative response in 30 patients before treatment. In addition, to investigate the mechanisms of action of interferon in promoting hepatitis B virus clearance, we serially measured the CD4/CD8 ratios, natural killer activity and lymphocyte proliferative response at wk 4, 8 and 12 of treatment. A beneficial response to therapy was defined as the sustained clearance of HBeAg and serum hepatitis B virus DNA within 1 yr of commencing therapy. Elevated IgM anti-HBc levels were associated with a beneficial response to therapy, but there was no correlation observed between response and pretreatment CD4/CD8 ratio, natural killer activity or lymphocyte proliferative response. Six of seven human leukocyte antigen DR3—positive patients responded. No measurable changes in the immunological parameters studied were observed in the nonresponder group, whereas a significant rise in CD4/CD8 ratio, associated with a fall in peripheral CD8 number and a decline in measurable NK activity, was seen in the responder group. These changes were maximal at the time of hepatitis B virus DNA clearance, which was associated with a transient increase in hepatic inflammation. Whether these changes occurred because of hepatic sequestration of cytotoxic and natural killer cells in the liver at the time of seroconversion or reflected an interferon-induced altered immunoregulatory balance requires further investigation. (HEPATOLOGY 1990;12:1111-1117).
Immunization of woodchucks with recombinant hepatitis delta antigen does not protect against hepatitis delta virus infection
Authors: Karayiannis Peter , Saldanha John , Monjardino John , Goldin Robert , Main Janice , Luther Shashi , Easton Mark , Ponzetto Antonio , Thomas Howard C.
Published: March 2007

To assess the role of immunization against hepatitis delta antigen in the prevention of hepatitis delta virus infection, woodchuck carriers of woodchuck hepatitis virus were immunized with a 64 amino acid portion of hepatitis delta antigen from its N-terminal region. The protein was expressed in Escherichia coli and contained a major immunogenic epitope. A significant anti-hepatitis delta response was observed that did not, however, protect the animals from hepatitis delta virus superinfection. Unexpectedly, the period of detectable viremia was longer in the immunized than in the control animals. We conclude that immunization with this recombinant hepatitis delta antigen does not afford protection against subsequent hepatitis delta virus exposure. (HEPATOLOGY 1990;12:1125-1128).
Antibodies to nuclear lamin C in chronic hepatitis delta virus infection
Authors: Węsierska-Gądek Józefa , Penner Edward , Hitchman Eva , Sauermann Georg
Published: March 2007

Sera of patients with chronic hepatitis delta virus infection stained the nuclear periphery in indirect immunofluorescence. Using proteins of isolated nuclei, isolated nuclear matrices, the nuclear pore complexlamina fraction and purified lamins A and C as antigen source in immunoblotting experiments, nuclear lamin C was identified as the reactive antigen. Most sera tested (8 of 10) recognized nuclear lamin C exclusively, but not the nuclear lamins A and B. Antibodies reacting with both nuclear lamins A and C, which share extensive sequence homologies, have been reported to occur in autoimmune hepatitis and primary biliary cirrhosis. The present findings suggest that the novel autoantibody associated with chronic hepatitis delta virus infection recognizes an epitope localized in the short carboxyterminal region of nuclear lamin C. (HEPATOLOGY 1990;12:1129-1133).
Endotoxin and the hyperdynamic circulation of portal vein—ligated rats
Authors: Mehta Rajeev , Gottstein Jeanne , Zeller Patrick W. , Lichtenberg Robert , Blei Andres T.
Published: March 2007

Humoral factors may be responsible for the hyperdynamic circulation seen in portal hypertension. Endotoxin, a peripheral arteriolar vasodilator, has been proposed to mediate this hemodynamic picture. We examined the pathogenic role of endotoxin in portal vein—ligated rats, a prehepatic portal hypertensive model with a well-developed hyperdynamic circulation. To this end, we (a) administered oral neomycin, a poorly absorbable antibiotic, at doses of 50 and 100 mg/day for 7 days and found no evident splanchnic hemodynamic effects of a 2-log—fold reduction of cecal aerobic bacterial flora as assessed by the radioactive microsphere technique in portal vein—ligated rats studied in the postanesthesia awake state; (b) assayed endotoxin in arterial samples using a quantitative limulus assay and found no evidence of endotoxinemia in PVL rats; (c) induced a state of endotoxin tolerance by repeated daily intraperitoneal injections of lowdose endotoxin and found no amelioration of the hyperdynamic state in portal vein—ligated rats. Our results do not support the hypothesis that endotoxin plays a major pathogenic role in the hyperdynamic circulation of this experimental model. (HEPATOLOGY 1990;12:1152-1156).
Calcium absorption in bone disease associated with chronic cholestasis during childhood†
Authors: Bucuvalas John C. , Heubi James E. , Specker Bonny L. , Gregg Deborah J. , Yergey Alfred L. , Vieira Nancy E.
Published: March 2007

Fractional absorption of calcium was determined in 9 children aged 4.9 to 16.9 yr with chronic cholestatic liver disease to determine the role of calcium malabsorption in the development of metabolic bone disease. Radiological evidence of rickets was absent in all patients, but bone density, measured by single beam photon absorptiometry of the distal radius, was reduced in eight of nine subjects. Serum calcium and phosphorus concentrations were normal in all except one subject. Serum 25-hydroxyvitamin D concentration was decreased compared with controls in only one of nine patients, but serum 1,25-dihydroxyvitamin D concentrations were diminished in seven of nine subjects. In all subjects, dietary calcium and phosphorus intakes were greater than 80% of the RDA. Fractional absorption of calcium, determined by oral and intravenous administration of stable calcium isotopes, was similar in cholestatic compared with control subjects (37.1% ± 12.5% vs. 34.0% ± 16.4%). In the cholestatic subjects, calcium absorption correlated with serum 1,25-dihydroxyvitamin D (r = 0.871, p < 0.002) but not 25-hydroxyvitamin D concentrations. Calcium balance, assessed by the duplicate diet method, was positive in four of five subjects. Anthropometric measurements were performed to examine the relationship between nutritional status and bone mineral content. Heights of all subjects were less than or equal to the 10th percentile and fat stores and somatic protein stores were less than the 25th percentile in six of nine subjects.

We conclude that factors other than calcium malabsorption and decreased serum 25-hydroxyvitamin D concentration contribute to diminished bone mass in children with cholestatic liver disease. (HEPATOLOGY 1990;12:1200-1205).
Defects in the precore region of the HBV genome in patients with chronic hepatitis B after sustained seroconversion from HBeAg to anti-HBe induced spontaneously or with interferon therapy
Authors: Takeda Kiyoshi , Akahane Yoshihiro , Suzuki Hiroshi , Okamoto Hiroaki , Tsuda Fumio , Miyakawa Yuzo , Mayumi Makoto
Published: March 2007

Hepatitis B virus DNA clones were propagated from sera of six patients with chronic hepatitis B who seroconverted from HBeAg to antibody to HBeAg either spontaneously or after administration of α-interferon. Defects in the precore region blocking synthesis and secretion of HBeAg were detected in all 46 hepatitis B virus DNA clones from three patients who remained positive for antibody to HBeAg and in whom hepatitis resolved. Defective clones had point mutations from guanine to adenine at nucleotide 83 in the precore region, converting codon 28 from tryptophan (TGG) to a stop codon (TAG). In contrast, this defect was not found in any of 39 hepatitis B virus DNA clones from three patients who seroconverted to antibody to HBeAg but then redeveloped HBeAg with reactivation of hepatitis. Using these results, the G-to-A point mutation at nucleotide 83 in the precore region would predict sustained positivity for antibody to HBeAg and remission of hepatitis in patients who have seroconverted either spontaneously or with interferon therapy. (HEPATOLOGY 1990;12:1284-1289).
Influence of hepatitis delta virus replication in the presence of hepatitis B virus DNA in peripheral blood mononuclear cells
Authors: Moraleda Gloria , Bartolomé Javier , Martinez Maria Gracia , Porres Juan Carlos , Carreño Vicente
Published: March 2007

The presence of hepatitis B virus DNA was studied in peripheral blood mononuclear cell samples from 259 HBsAg carriers (229 anti-hepatitis delta negative, 30 anti-hepatitis delta positive), 16 anti-HBc—positive HBsAg-negative patients and 30 patients without hepatitis B virus markers. Hepatitis B virus DNA sequences were detected in peripheral blood mononuclear cell from 115 (44.4%) of the chronic HBsAg carriers and from two (12%) of the anti-HBc—positive, HBsAg-negative patients. In anti-hepatitis delta—negative patients, viral DNA was positive in peripheral blood mononuclear cell from 74 (46%) and from 24 (35.5%) with and without serum HBV-DNA, respectively. With respect to anti-hepatitis delta—positive patients, viral DNA was found in peripheral blood mononuclear cell in 8 of 13 (61.5%) of the patients with circulating hepatitis delta virus RNA and in 9 of 17 (53%) of the hepatitis delta virus RNA—negative subjects. Regarding hepatitis B virus DNA in serum and peripheral blood mononuclear cell, 71% (5 of 7) of the patients with serum hepatitis B virus DNA had this marker in peripheral blood mononuclear cell, whereas 52% (12 of 23) of the patients without serum hepatitis B virus DNA had hepatitis B virus DNA in peripheral blood mononuclear cell. A Southern blot analysis was also carried out on peripheral blood mononuclear cell samples from 30 patients. Hepatitis B virus DNA was detected in 16 patients as free forms, in 12 patients as dimers and free forms and as free circular together with free linear forms in the remaining two patients. In conclusion, there is no relation between the viral replication level and the presence of hepatitis B virus DNA in peripheral blood mononuclear cell. Hepatitis delta virus does not seem to affect the presence of hepatitis B virus DNA in peripheral blood mononuclear cell. (HEPATOLOGY 1990;12:1290-1294).
Primary culture of adult rat hepatocytes after 48-hour preservation of the liver with cold UW solution
Authors: Guyomard Claire , Chesne Christophe , Meunier Bernard , Fautrel Alain , Clerc Catherine , Morel Fabrice , Rissel Maryvonne , Campion Jean-Pierre , Guillouzo André
Published: March 2007

Rat livers were perfused and stored for 48 hr in cold University of Wisconsin solution before dissociation by the two-step collagenase method. At that time, glycogen content was significantly reduced, but no obvious changes in albumin, β-actin and aldolase B mRNAs and in glutathione levels were observed. Enzymatic perfusion yielded 280 ± 30 × 106 viable hepatocytes vs. 520 ± 40 × 106 viable hepatocytes from unstored organs. Cell viability determined by trypan blue exclusion was 74% and 90%, respectively. Hepatocytes from University of Wisconsin—preserved livers had a 29% reduced adenosine triphosphate content, but glutathione levels did not significantly differ from those found in unstored cells. When put into culture, hepatocytes formed typical monolayers of granular epithelial cells and did not exhibit alteration of their fine structure when compared with cells from unstored organs. After 24 and 48 hr, they showed variations in cytochrome P-450 content and ethoxyresorufin O-deethylase activity similar to those observed with unstored cells. By contrast, overall protein synthesis and albumin secretion rate were 40% and 30% lower, respectively. Hepatocytes from University of Wisconsin—preserved organs could be cryopreserved and further cultured as unstored cells. The University of Wisconsin solution was also used to preserve isolated hepatocytes. Viability of freshly isolated hepatocytes was decreased by only 10% after 48 hr of hypothermic liver storage when assayed by intracellular lactate dehydrogenase content. However, after 4 hr of storage, in contrast with hepatocytes preserved in L15 Leibovitz medium, the cells attached poorly to plastic and exhibited morphological alterations. These results show that rat hepatocytes are still viable, well functional and able to survive in culture after 48 hr University of Wisconsin preservation of the liver and suggest that isolated hepatocytes survive better in a culture medium than in this organ storage solution. (HEPATOLOGY 1990;12:1329-1336).
Serial quantitative image analysis and confocal microscopy of hepatic uptake, intracellular distribution and biliary secretion of a fluorescent bile acid analog in rat hepatocyte doublets
Authors: Kitamura Tsuneo , Gatmaitan Zenaida , Arias Irwin M.
Published: March 2007

To characterize the poorly understood mechanisms of intracellular transport of bile acids, fluorescein isothiocyanate—glycocholate was synthesized and its ring-OH—linked structure established by fast atom bombardment, mass spectroscopy and 13C nuclear magnetic resonance. Biliary secretion of fluorescein isothiocyanate—glycocholate and [14 C]-labeled glycocholate in rats was similar, in contrast to the biliary secretion of sodium fluorescein and methylamine-conjugated fluorescein isothiocyanate, which are nonbile acid organic anions. After incubation of cultured hepatocyte doublets with fluorescein isothiocyanate—glycocholate, serial quantitative image analysis of fluorescence was performed in the cellular cytoplasm, perinuclear zone and bile canaliculus. Uptake of fluorescein isothiocyanate—glycocholate into the cytoplasm was inhibited by removal of sodium from the medium and by addition of glycocholate or taurocholate. After preincubation with colchicine, but not lumicolchicine, the proportion of perinuclear to cytoplasmic fluorescein isothiocyanate—glycocholate increased during incubation with fluorescein isothiocyanate—glycocholate. Neither fluorescein isothiocyanate—glycocholate uptake nor canalicular secretion was affected. By confocal microscopy the perinuclear zone containing fluorescein isothiocyanate—glycocholate was identified as the Golgi apparatus by fluorescent colocalization with C6-NBD-ceramide, which specifically identifies the Golgi apparatus. We conclude that colchicine inhibits fluorescein isothiocyanate—glycocholate transfer from the Golgi apparatus to the bile canaliculus. These results suggest that intracellular bile acid transport involves microtubule-dependent vesicular movement from the Golgi apparatus to the bile canaliculus. The role of this process in normal physiology is uncertain. (HEPATOLOGY 1990;12:1358-1364).
Differential regulation of liver P-450III cytochromes in choline-deficient rats: Implications for the erythromycin breath test as a parameter of liver function
Authors: Kolars Joseph C. , Murray Scott A. , Peters Ken M. , Watkins Paul B.
Published: March 2007

Progressive liver fibrosis in rats develops when they are fed a diet deficient in choline. This diet also results in a pronounced and selective decrease in the liver microsomal content of a phase I drug—metabolizing enzyme belonging to the cytochrome P-450III gene family. Because P-450III cytochromes characteristically catalyze the N-demethylation of erythromycin, we believed that the production of breath CO2 from erythromycin would be dramatically reduced in choline-deficient rats. However, when 12 choline-deficient rats were compared with 9 control rats, the reduction in CO2 production from erythromycin (mean decrease 71%) was essentially identical to that from aminopyrine (mean decrease 69%), a substrate believed to be metabolized normally by the hepatocyte in fibrotic liver disease. Furthermore, we found that the relative erythromycin and aminopyrine demethylase activities were comparable when measured in vitro in liver microsomes prepared from the choline-deficient rats. To determine the molecular basis for the erythromycin demethylase activity in the choline-deficient rats, the liver microsomes were subjected to immunoblot analysis using a variety of polyclonal and monoclonal antibodies capable of distinguishing individual P-450III—related proteins. Our studies confirm that a major erythromycin demethylase belonging to the P-450III family, termed P-450p, was greatly reduced in the choline-deficient rat liver. However, the specific concentration of a second P-450p—related protein was essentially normal and that of a third P-450p—related protein was actually increased in the choline-deficient rat liver. These changes occurred over weeks and months on the choline-deficient diet and are not consistent with “feminization” of the liver. The P-450p—related proteins appeared to catalyze erythromycin demethylase activity because antibodies recognizing them inhibited the majority (62%) of this activity in choline-deficient rat microsomes. Finally, RNA extracted from choline-deficient rat livers was hybridized on Northern blots with synthetic oligonucleotide probes that identify the only two known rat P-450III cDNAs, PCN-1 and PCN-2. Prolonged choline deficiency appeared to have no consistent effects on expression of these RNA species. We conclude that despite the down-regulation of a major erythromycin demethylase, this catalytic activity is relatively preserved in the choline-deficient rat, at least in part because of differential regulation of P-450III cytochromes. Erythromycin therefore appears to offer little advantage over aminopyrine as a substrate for assessing liver function in this model of chronic liver disease. (HEPATOLOGY 1990;12:1371-1378).
Hepatobiliary effects of morphine are mediated in the brain
Authors: Hurwitz Aryeh , Looney Greg , Sullins Mark , Ben-Zvi Zvi
Published: March 2007

Morphine slows hepatobiliary elimination of sulfobromophthalein in rodents, raising dye levels in plasma and liver. Earlier studies showed these effects to be independent of other opiate effects such as bile duct spasm, hypothermia or blood gas changes resulting from respiratory depression. Because opiate receptors are distributed throughout the body, within the central nervous system and at peripheral sites including the gastrointestinal tract, experiments were performed to ascertain whether central or peripheral sites mediate the hepatobiliary effects of morphine. Sulfobromophthalein was administered intravenously to mice and its levels were measured in plasma and liver. Tail-flick latency indicated centrally mediated analgesia. Inhibited intestinal transit of India ink reflected an opiate effect with a significant peripheral component.

When injected into a cerebral ventricle morphine was much more potent in producing analgesia and raising sulfobromophthalein levels than when administered intravenously or intraperitoneally. An intravenous dose of naloxone that reversed morphine analgesia also prevented sulfobromophthalein elevation but did not prevent gut slowing. Naltrexone injected in a cerebral ventricle also reversed analgesia and sulfobromophthalein elevation but not intestinal slowing. The polar opiate agonist N-methylmorphine did not cause analgesia or raise sulfobromophthalein levels at peripheral intraperitoneal doses to 100 mg/kg. When given in a central ventricle at 4 × 10—3 mg/kg, this agent produced analgesia and raised sulfobromophthalein but did not slow intestinal transit. After spinal cord transection, intravenous morphine did not retard the tail-flick response or affect sulfobromophthalein disposition, but peripherally mediated intestinal transit was slowed as it was in intact mice.

These experiments demonstrate parallel opiate effects on analgesia and on BSP disposition but not on intestinal transit. Hepatobiliary effects of morphine are mediated by interaction with opiate receptors in the brain. (HEPATOLOGY 1990;12:1406-1412).