bluuebunny/crossref_metadata_2025 huggingface dataset:
Blood-brain barrier permeability is markedly decreased in cholestasis in the rat
Authors: Wahler Jacqueline B. , Swain Mark G. , Carson Richard , Bergasa Nora V. , Jones Anthony E.
Published: March 2007

The blood-brain-barrier plays an essential role in regulating the entrance of substances into the brain. To date, permeability of the blood-brain barrier has not been studied in models of cholestatic liver injury, although levels of substances known to enhance vascular permeability (bile acids, substance P, histamine) are elevated in cholestasis. Two rat models of cholestasis were studied: bile duct resection (5 days after surgery) and α-naphthylhisothiocyanate treatment (45 mg/kg/day for 7 days). The mean value for whole brain blood-to-brain transfer constant in bile duct resection rats was about 50% less than corresponding values in sham-operated and unoperated control rats (p ± 0.05, respectively). Reductions in blood-to-brain transfer constant of similar magnitude were found in the caudate nuclei, cortexes and hippocampi of bile duct-resected rats. Blood-to-brain transfer constant values in α-naphthylisothiocyanate-treated rats were also about 50% less in whole brain and specific brain regions than corresponding control values. A precedent for a decrease in blood-to-brain transfer constant is the dexamethasone-treated rat, in which the phenomenon has been attributed to a decrease in cerebral capillary endothelial cell membrane fluidity. We confirmed that blood-to-brain transfer constant values are reduced by about 50% in dexamethasone-treated rats. A decrease in membrane fluidity affords a rational explanation for a decrease in blood-to-brain transfer constant in cholestasis as a consequence of the dynamic equilibrium between elevated plasma levels of cholesterol in cholestasis and cell membranes exposed to the circulation. (Hepatology 1993;17:1103-1108.)
Artificial liver: A forthcoming attraction
Authors: Sussman Norman L. , Kelly James H.
Published: March 2007

We have developed a novel hepatocyte loaded hollow fiber bioreactor as a potential bioartificial liver. Freshly harvested rat hepatocytes were entrapped in a three-dimensional gel matrix within hollow fibers in a perfused bioreactor. Gel entrapment allowed cells to be cultured at high density while maintaining tissuespecific function. Hepatocyte function was evaluated in 10 bioreactors, each containing approximately 5 x 107 cells. Oxygen consumption averaged 0.32 pmole/cell/hr, albumin appearance averaged 0.60 pg/cell/hr, and lidocaine clearance (a measure of the P-450 activity) averaged 0.74 pg/cell/hr. Function persisted for the 7 days of the study. Electron microscopy at 7 days showed the distinctive ultrastructure of viable, differentiated hepatocytes: bile canaliculi, intercellular junctions, peroxisomes, abundant mitochondria, and glycogen granules. Maintenance of tissue specific function and ultrastructure suggests that this bioreactor configuration has potential as a device to support patients in liver failure, as well as to study hepatocytes in vitro.
Quantification of hepatitis C virus by competitive reverse transcription—polymerase chain reaction: Increase of the virus in advanced liver disease
Authors: Kato Naoya , Yokosuka Osamu , Hosoda Kazuhiko , Ito Yoshimi , Ohto Masao , Omata Masao
Published: March 2007

We developed a quantitative method of hepatitis C virus RNA by competitive reverse transcription-polymerase chain reaction. With this method, 36 patients with type C chronic liver disease were analyzed for the copy number of circulating hepatitis C virus in 50 μl of serum. The amounts of hepatitis C virus RNA ranged from 101 to 107 copies in the 36 patients. The average amount of hepatitis C virus RNA was 103.3 ± 2.2 copies in 12 patients with chronic persistent hepatitis, 105.7 ± 1.6 copies in 12 patients with chronic active hepatitis and 106.0 ± 1.6 copies in 12 patients with cirrhosis (including 4 patients with hepatocellular carcinoma). The amount of hepatitis C virus RNA in serum was significantly less in patients with chronic persistent hepatitis than in patients with chronic active hepatitis or cirrhosis (p <0.01), and it tended to increase according to the progression of histopathological changes of the liver. Furthermore, it was revealed that the amount of hepatitis C virus RNA became exponentially larger as the term from infection became longer. Quantification of hepatitis C virus RNA by competitive reverse transcription-polymerase chain reaction may have many applications for the study of clinical features of hepatitis C virus infection. (Hepatology 1993;18:16-20).
Circulating and tissue levels of the neutrophil chemotaxin interleukin-8 are elevated in severe acute alcoholic hepatitis, and tissue levels correlate with neutrophil infiltration
Authors: Sheron Nick , Bird George , Koskinas John , Portmann Bernard , Ceska Miroslav , Lindley Ivan , William Roger
Published: March 2007

Acute alcoholic hepatitis is characterized by a unique degree of liver neutrophil infiltration, often accompanied by marked peripheral neutrophilia in the absence of demonstrable bacterial or fungal infection. In this study we assayed plasma and tissue levels of a potent neutrophil activator and chemotaxin, interleukin-8, in patients with a spectrum of alcoholic liver diseases and in normal and diseased control subjects. Levels of circulating interleukin-8 were undetectable in normal subjects but highly elevated in patients with alcoholic hepatitis, particularly in those who died (geometric mean = 600 ng/L confidence interval = 323 to 1,120 vs. geometric mean = 184 ng/L confidence interval = 114 to 309 in survivors). Levels correlated with biochemical indicators of severe disease (bilirubin: R = 0.38; international prothrombin ratio: R = 0.28; white blood cell count: R = 0.35; creatinine: R = 0.34) and with tumor necrosis factor-α (R = 0.43) and soluble tumor necrosis factor receptors (p55; R = 0.59). In contrast, moderate elevations in the levels of circulating interleukin-8 were seen in alcoholic cirrhosis (geometric mean = 93 ng/L confidence interval = 40 to 213) and in alcoholic patients undergoing alcohol withdrawal (geometric mean = 137 ng/L confidence interval = 72 to 259). Levels in nonalcoholic inflammatory liver disease were comparatively low (geometric mean = 17 ng/L confidence interval = 10 to 29). In liver tissue from patients with alcoholic liver disease, local levels of interleukin-8 correlated with degree of neutrophil infiltration (R = 0.71; n = 16), and levels were much higher in alcoholic hepatitis (987 pg/mg confidence interval = 351 to 1623) than in other alcoholic liver diseases (103 pg/mg confidence interval = 0 to 220), normal liver (20 pg/mg confidence interval = 0 to 61) and nonalcoholic liver disease (219 pg/mg confidence interval = 142 to 295). Immunohistochemical study confirmed widespread parenchymal expression of interleukin-8 in alcoholic hepatitis patients with the most severe degrees of neutrophil infiltration. Interleukin-8 may play a role in mediating the infiltration of neutrophils and the effects of neutrophil activation so characteristic of acute alcoholic hepatitis. (Hepatology 1993;18:41-46).
Terlipressin plus transdermal nitroglycerin vs. octreotide in the control of acute bleeding from esophageal varices: A multicenter randomized trial
Authors: Silvain Christine , Carpentier Stéphane , Sautereau Denis , Czernichow Bella , Métreau Jean-michel , Fort Eric , Ingrand Pierre , Boyer Jean , Pillegand Bernard , Doffël Michel , Dhumeaux Daniel , Beauchan Michel
Published: March 2007

We undertook a multicenter randomized trial to compare the efficacy of terlipressin combined with transdermal nitroglycerin and that of octreotide in the emergency control of acute variceal hemorrhage in cirrhosis. Over 16 mo, 87 patients with endoscopically proved active bleeding from esophageal or cardiac varices were enrolled in five centers in France and randomly assigned to receive intravenous terlipressin (2 mg and then 1 mg/4 hr over 24 hr) and transdermal nitroglycerin (10 mg/12 hr over 24 hr) (group 1) or octreotide (continuous intravenous infusion of 25 μg/hr over 12 hr and then 100 μg at hr 12 and hr 18 subcutaneously) (group 2). Initial control of bleeding was assessed at the end of 12 hr of treatment on the basis of stability of blood pressure and hematocrit level with no further transfusion requirements. At 12 hr, bleeding was controlled in 59% (24 of 41) in group 1 and 78% (36/46) of group 2 patients (Fisher's exact test, p = 0.064). Mean transfusion requirements over this 12-hr period were significantly greater in group 1 (three blood units; range = 0 to 13) than in group 2 (one blood unit; range = 0 to 5) (p = 0.002). After the first 12 hr, 20% of patients (5 of 24) had repeat bleeding in group 1 compared with 27% (10 of 36) in group 2. During the first 48-hr period, five patients (12%) died in group 1, compared with 3 (6%) in group 2. Few side effects were noted in either group. However, in group 1 two patients experienced severe bradycardia; it resulted in death in one patient. Octreotide appeared to be as effective as terlipressin combined with transdermal nitroglycerin in the emergency control of active variceal bleeding in cirrhosis, with significantly smaller transfusion requirements and only minor side effects. (Hepatology 1993;18:61-65).
Prior esophageal variceal bleeding does not adversely affect survival after orthotopic liver transplantation
Authors: Ho Ka-Sic , Lashner Bret A. , Emond Jean C. , Baker Alfred L.
Published: March 2007

Prior variceal bleeding may adversely affect the prognosis of orthotopic liver transplantation. We studied this question by evaluating all 175 adult patients undergoing orthotopic liver transplantation at our institution to determine risk factors associated with mortality after transplantation. Seventy patients demonstrated prior variceal bleeding, and of those, 32 had a course of sclerotherapy. Thirteen also had portal systemic shunts. Compared with the 105 transplant patients who had no prior bleeding, patients who bled were more likely to have parenchymal liver disease (74% vs. 50%), equally likely to have cholestatic liver disease (19%), and less likely to have malignancy (6% vs. 19%) or fulminant liver failure (1% vs. 12%). Bleeding patients also were more likely to be Child-Pugh class C (46% vs. 35%). By the common closing date of December 31, 1990, 26 patients (37%) with prior bleeding and 48 patients (46%) without bleeding died after transplantation. From survival curves, patients with prior bleeding had improved survival rates at 1 yr (65% vs. 54%), at 3 yr (60% vs. 47%) and at 5 yr (55% vs. 43%), although the differences were not statistically significant. With a proportional hazards model to adjust for confounding effects, the relative risk of mortality among patients with prior bleeding was 0.60 (95% confidence interval, 0.27 to 1.30). Sclerotherapy or portal systemic shunts did not alter survival. Important adverse risk factors for mortality included older age at transplantation, black race, malignant disease and more advanced Child-Pugh class. This study suggests that prior esophageal variceal bleeding did not adversely affect the mortality of patients undergoing orthotopic liver transplantation. (Hepatology 1993;18:66-72).
Normal splanchnic but impaired peripheral insulin-stimulated glucose uptake in cirrhosis
Authors: Shmueli Ehoud , Walker Mark , Alberti George , Record Christopher O.
Published: March 2007

The site of impaired glucose uptake in cirrhosis is uncertain. We therefore performed hyperglycemic clamps (glucose 10 mmol/L) in 10 patients with compensated alcoholic cirrhosis and impaired glucose tolerance and in six control subjects. Muscle glucose uptake was estimated in patients and controls with the forearm technique. In the cirrhotic subjects splanchnic glucose uptake was measured with hepatic vein catheterization and primed continuous infusions of indocyanine green and [6−3H]glucose. To assess insulinindependent glucose uptake and the effects of elevated nonesterified fatty acid levels on glucose uptake, we repeated the study with somatostatin to induce insulin deficiency and a nicotinic acid analog, acipimox, to inhibit lipolysis. Substrate disposal was assessed on indirect calorimetry. Despite similar stimulated insulin levels, total glucose utilization was lower in the cirrhotic subjects (3.9 ± 0.3 vs. 8.8 ± 1.7 mg/kg/min, p = 0.006). This deficiency was accounted for by lower nonoxidative glucose disposal (1.2 ± 0.2 vs. 5.8 ± 1.6 mg/kg/min, p = 0.002). Forearm glucose uptake was lower in the cirrhotic subjects (0.39 ± 0.06 vs. 1.21 ± 0.3 mg/100 ml/min, p = 0.001). However, splanchnic glucose uptake at 1.59 ± 0.14 mg/kg/min was similar to that reported in other studies of normal subjects. Insulin-independent glucose uptake was normal, and acipimox had no effect on total or forearm glucose utilization. Glucose intolerance in cirrhosis is characterized by impaired peripheral insulin-stimulated nonoxidative glucose disposal. The high nonesterified fatty acid levels seen in cirrhosis most likely do not contribute to this defect. Splanchnic glucose uptake is normal in cirrhosis. (Hepatology 1993;18:86-95).
Can determination of the proliferative capacity of the nontumor portion predict the risk of tumor recurrence in the liver remnant after resection of human hepatocellular carcinoma?
Authors: Chiu Jen-Hwey , Wu Li-Hwa , Kao Hwa-Li , Chang Hwey-May , Tsay Shyh-Haw , Loong Che-Chuan , Cha Gar-Yang , Lui Wing-Yu
Published: March 2007

To test the hypothesis that increased proliferative capacity of cells in a liver remnant is a risk factor for tumor recurrence in patients who have undergone liver resection for hepatocellular carcinoma, DNA flow-cytometric measurement and cell-cycle analysis of the nontumor parts of resected hepatocellular carcinomas (tumor size <5 cm) were performed. The disease-free survival rates 1, 2, 3 and 4 yr after surgery were 64%, 58%, 43%, and 36%, respectively. Proliferative capacity (fractions of synthetic, postsynthetic and mitotic phases) of the nontumor parts, irrespective of liver pathology, was higher than that of normal liver and statistically lower than that of tumor parts from resected hepatocellular carcinoma specimens. Livers with chronic active hepatitis and with hepatocyte dysplasia had significantly lower proliferative activity than did those with chronic active hepatitis and with hepatocyte dysplasia. We saw no significant difference in proliferative capacity between patients with and without cirrhosis. Disease-free-survival analysis showed that the presence of liver pathology (hepatitis B infection, cirrhosis, chronic active hepatitis and hepatocyte dysplasia) was not the factor linked to tumor recurrence in the liver remnant and that a marked increase in proliferative capacity (≥ 18%), regardless of liver pathology, was the risk factor linked to tumor recurrence after liver resection. We conclude that there is some degree of increased proliferative capacity in the nontumor parts of resected hepatocellular carcinomas and that a marked increase in the proliferative capacity (≥ 18%) of the nontumor part is a significant risk factor in predicting tumor recurrence in the liver remnant after liver resection. (Hepatology 1993;18: 96-102).
Decrease of hepatic triglyceride lipase levels and increase of cholesteryl ester transfer protein levels in patients with primary biliary cirrhosis: Relationship to abnormalities in high-density lipoprotein
Authors: Hiraoka Hisatoyo , Yamashita Shizuya , Matsuzawa Yuji , Kubo Masaharu , Nozaki Shuichi , Sakai Naohiko , Hirano Ken-Ichi , Kawata Sumio , Tarui Seiichiro
Published: March 2007

Serum levels of high-density lipoprotein cholesterol are often increased in patients with primary biliary cirrhosis. To elucidate the mechanism of the elevation of high-density lipoprotein cholesterol levels in this disease, lipoprotein abnormalities were analyzed in 10 patients subdivided into two groups according to concentration of high-density lipoprotein cholesterol. Activities and protein masses of lipoprotein lipase, hepatic triglyceride lipase and cholesteryl ester transfer protein were also determined. Serum highdensity lipoprotein cholesterol concentration exceeded 90 mg/dl in 5 of 10 patients. Lipoprotein particles in the high-density lipoprotein2 fraction (with density between 1.063 and 1.125 gm/ml) were enriched with apolipoprotein E and larger in size than those of normal controls. In patients with and without hyperalphalipoproteinemia, hepatic triglyceride lipase activity was significantly decreased (p < 0.05); this was due to the reduction of its protein mass. Lipoprotein lipase activity and protein mass were normal. It is noteworthy that increases in cholesteryl ester transfer protein activity and mass were observed. The enhancement of cholesteryl ester transfer protein activity was more remarkable in the patients with hyperalphalipoproteinemia than in those without hyperalphalipoproteinemia. Because a significant postive correlation was demonstrated between activity and protein mass (r = 0.90, p <0.001), the increase of cholesteryl ester transfer protein activity may be attributed to the increase of its protein mass. These data suggest that the decrease of hepatic triglyceride lipase levels at least partly explains the appearance of apolipoprotein E-rich large high-density lipoprotein particles in patients with primary biliary cirrhosis. Considering that the deficiency of cholesteryl ester transfer protein has been shown to cause marked hyperalphalipoproteinemia with similar abnormalities of high-density-lipoprotein particles, the increase of cholesteryl ester transfer protein activity in primary biliary cirrhosis with hyperalphalipoproteinemia is peculiar and interesting, although its mechanism remains unknown. (Hepatology 1993;18:103-110).
The immunosuppressive effect of ursodeoxycholic acid: A comparative in vitro study on human peripheral blood mononuclear cells
Authors: Lacaille Florence , Paradis Khazal
Published: March 2007

Ursodeoxycholic acid is an efficient treatment for putatively immune-mediated liver diseases, but its mechanism of action is unknown. We studied human mononuclear cell proliferation as an in vitro model for cell-mediated immunity in the presence of ursodeoxycholic acid, its glycoconjugate and tauroconjugate and chenodeoxycholic acid at concentrations of 5, 25 and 50 μmol/L. Proliferation was inhibited in a dose-dependent manner compared with control values (15% to 54% depending on the bile acid, concentration and mitogen used), except at 5 μmol/L where inhibition was significant with only one mitogenic stimulus of the three used. With one mitogen (phorbolester) the inhibition was additive with that of cyclosporine. The number of cell-surface receptors studied was not modified by bile acids. Interleukin-2 production was decreased 35% to 60% by ursodeoxycholic acid and its conjugates. The proliferation of the interleukin-2-dependent cell line CTLL-2 was also inhibited. The immunosuppression was reversible except at a chenodeoxycholic acid concentration of 50 μmol/L. Because bile acids are able to partition into membranes and change their properties, we speculate that this allows them to interact with cell-surface receptors or signaling systems within the membrane or on its inner face, thus impairing their function. This would inhibit the numerous extracellular messages that lymphocytes need to proliferate. (Hepatology 1993;18:165-172).
Reestablishment of cell polarity of rat hepatocytes in primary culture
Authors: Musat Alexandru I. , Sattler Carol A. , Sattler Gerald L. , Pitot Henry C.
Published: March 2007

The cell-basement membrane interaction is an important determinant of epithelial cell polarity. Although hepatocytes in situ are polarized, no morphologically identifiable basement membrane is found at their basal surface. However, several studies have demonstrated immunoreactivity to basement membrane proteins in the space of Disse, indicating the existence of an extracellular matrix, albeit of low density. Therefore we hypothesized that the interaction of hepatocytes with this matrix may determine their polarity and asked whether a basement membrane-like substrate could reestablish hepatocyte polarity in vitro. For this purpose, established monolayers of primary rat hepatocytes were cultured overlaid with a basement membrane-like matrix extracted from the Engelbreth-Holm-Swarm mouse tumor, mimicking thus the in situ tissue architecture. The hepatocytes in this culture configuration, unlike hepatocytes in classic cultures, developed distinct membrane domains, as demonstrated by the reformation of γ-glutamyltranspeptidase, Mg2+-ATPase-positive bile canalicular networks and intercellular gap junctions immunolocalized to the lateral membrane with antibodies to connexin 32. The actin cytoskeleton of these cells reorganized into pericanalicular webs, and no accumulation of “stress” filaments was found beneath the membrane facing the medium. Golgi complexes appeared to be preferentially located in mitochondria-poor pericanalicular cytoplasm, indicating the polarized distribution of these organelles. Together, these data indicate that a basement membrane-like substrate present between hepatocytes and nutrient medium restores the polarity of these cells in culture. Extrapolation of these findings to the intact liver suggests that the matrix in Disse's space governs the development of hepatocyte polarity. (Hepatology 1993;18:198-205).
Restructuring American health care financing: First of all, do no harm!
Authors: Berk Paul D.
Published: March 2007

Health care costs are climbing throughout the western world. Aging populations and the costs of advanced technology are the principal forces behind much of this global increase. No country has yet succeeded in containing these growing costs other than by some form of rationing. A variety of experimental strategies, including managed competition, are being considered or tested, but none is clearly effective.

American health care expenditures differ, not in that they are rising, but in their enormously high starting point. Among other things, our higher costs reflect administrative costs of more than 20%, double those of Canada and nearly triple the European average; a malpractice system that, whatever its possible advantages, costs more than 10 times as much as it pays out to the injured; the enormous medical costs of poverty; maldistribution of physician specialties and incomes; and reimbursement systems that eliminate consumer input and oversight.

Restructuring the system of health care financing to bring administrative costs in line with those of other nations could save at least $70 billion annually; another $25 billion or more could be saved by replacing the malpractice system with more cost-effective alternatives. These savings could defray the costs of insuring all those not now covered, without increasing either costs to the middle class, through taxation of benefits, or total health care expenditures. With all Americans covered, the necessary restructuring of the system of health care delivery could be conducted without the current pressure for immediate drastic reform, which carries with it the risk of serious error.

In dealing with the sick, physicians are taught to apply two maxims: “primum non nocere” or “first of all, do no harm!”; and the rule of therapeutic restraint. The latter states that a severe chronic illness may respond better, and with fewer complications, to gradual corrective measures than to highly aggressive therapy. Both rules could well be applied to curing the American health care system.
Liver transplantation for hepatocellular carcinoma
Authors: Venook Alan P.
Published: March 2007

Background: Liver transplantation for unresectable hepatocellular carcinoma yields disappointing results. Most cases recur within 2 years, often in the transplanted liver. Methods: A combination of neoadjuvant doxorubicin and orthotopic liver transplantation was used in 20 patients with unresectable hepatocellular carcinoma confined to the liver. Seventeen patients had tumors > 5 cm in greatest diameter, and 11 cases were stage IVA by the TNM classification. Doxorubicin was administered preoperatively, intraoperatively, and postoperatively at a dose of 10 mg/m2 weekly, totaling 200 mg/m2. Results: Chemotherapy was well tolerated although leukopenia was was observed in 70% of patients. Eight patients died, five of recurrent tumor and three of hepatitis B. Three others remain alive 8-22 months after tumor recurrence. One patient had initial tumor recurrence in the allograft. Actuarial survival is 59% and tumor-free survival is 54% at 3 years. For the 17 patients with tumors > 5 cm, overall survival is 63% and tumor-free survival is 49% at 3 years. Conclusion: The results of this pilot study suggest that neoadjuvant doxorubicin chemotherapy favorably alters the post-transplant survival of patients with ehpatocellular carcinoma.
Hepatitis C virus c100 antigen in liver tissue from patients with acute and chronic infection
Authors: Sansonno Domenico , Dammacco Franco
Published: March 2007

A pool of murine monoclonal antibodies developed against c100 antigen, a hepatitis C virus-associated protein encoded by the NS3/NS4 virus genome, was used to detect hepatitis C virus in liver biopsy specimens from patients with acute and chronic hepatitis C virus infection. The antigen was present in the cytoplasm of liver cells only. The immunoreactive signal appeared as large, distinct, brilliant fluorescent granules with no clear relationship to cellular structures. No obvious membrane c100 antigen accumulation was observed. Distribution of c100-containing hepatocytes was directly correlated with viral replication in acute hepatitis. All three acute-hepatitis patients were positive for hepatitis C virus RNA (as detected on polymerase chain reaction) in serum and displayed c100 antigen in 50% to 70% of hepatocytes, with a distinct topographical relationship with necrotic areas and inflammatory cell accumulation. Conversely, very low numbers of infected cells and no relationship between tissue c100 antigen expression and sites of liver cell necrosis and inflammation were found in 14 chronic hepatitis C virus infection patients. Furthermore, though all patients had measurable levels of serum hepatitis C virus RNA, only eight (57%) had detectable c100 antigen in liver sections. Indeed, these two distinct immunopathological patterns were inversely related to the development of c100 antibody in serum. Specificity of hepatocellular c100 antigen deposits was established through extensive absorption experiments using structural and nonstructural hepatitis C virus recombinant proteins. However, tissue processing was found to be a crucial step in the demonstration of hepatitis C virus antigen in fresh frozen liver tissue. Furthermore, monoclonal antibodies against recombinant core and NS3-encoded proteins failed to demonstrate immunofluorescent staining, suggesting that the c100 protein is the main antigen demonstrable by this method. (Hepatology 1993;18:240-245).
Adoptive transfer of immunity to hepatitis B virus after T cell-depleted allogeneic bone marrow transplantation
Authors: Ilan Yaron , Nagler Arnon , Adler Ruth , Naparstek Elizabeth , Or Reuven , Slavin Shimon , Brautba Chaim , Shouva Daniel
Published: March 2007

Recipients of allogeneic bone marrow transplantation are pancytopenic for several weeks and immunosuppressed for many months as a result of myeloablative therapy required to eliminate the basic disease and to prevent allograft rejection. After bone marrow transplantation, these patients remain profoundly immunosuppressed by the chemotherapy and immunotherapy used as prophylaxis against graft-vs.-host disease, treatment of established disease or both. These patients are usually dependent on multiple blood products and are therefore at risk for hepatitis B virus infection, which may run a fulminant course. Active immunization against hepatitis B virus in the immediate pre-bone marrow transplantation and post-bone marrow transplantation periods was found to be ineffective, probably because of the absence of T cell-dependent B-cell responses, which persists for approximately 1 yr after bone marrow transplantation. We studied adoptive transfer of immunity to hepatitis B virus through bone marrow transplantation in two populations of patients. The first group (A) consisted of 12 pairs of BMT donors and recipients, in which all bone marrow donors were positive for antibodies to HBc and HBs as a result of previously acquired hepatitis B virus infection and resolution; all recipients were negative to antibodies to HBc and HBs. The second group (B) consisted of eight pairs of donors and recipients in which all the donors were actively immunized against hepatitis B virus before bone marrow transplantation; all recipients were negative for all hepatitis B virus markers. All bone marrow transplantation recipients were monitored for antibodies to hepatitis B virus antigens. In group A, antibodies to HBsAg and HBcAg were detected 30 to 120 days after bone marrow transplantation. In group B, all immunized bone marrow transplantation donors demonstrated seroconversion until the day of bone marrow transplantation. In group B bone marrow transplantation recipients, protective HBs antibody levels appeared in all eight patients 9 to 42 days after bone marrow transplantation. This study provides evidence that primary immunization of bone marrow donors against hepatitis B may induce antibodies to HBsAg in bone marrow recipients through adoptive transfer of immunity. Moreover, bone marrow transplantation recipients may respond to booster vaccination after primary immunization of the bone marrow transplantation donor. (Hepatology 1993;18:246-252).
High prevalence of thyroid autoantibodies in a prospective series of patients with chronic hepatitis C before interferon therapy
Authors: Tran Albert , Quaranta Jean François , Benzaken Sylvia , Thiers Valerie , Chau Hoang Trong , Hastier Patrick , Regnier Daniel , Dreyfus Gilles , Pradier Christian , Sadoul Jean-Louis , Hebutern Xavier , Rampa Patrick
Published: March 2007

After describing two cases of Hashimoto's thyroiditis associated with chronic hepatitis C, we set up a prospective study to assessthe prevalence of thyroid autoantibodies (thyroglobulin and thyroid microsomal autoantibodies) in 72 chronic hepatitis C patients (43 men and 29 women; mean age = 51 ± 2.1 yr) before interferon therapy admitted between January and December 1991 to our liver unit. Thyroid autoantibodies were systematically assayed in 60 chronic HBsAg-positive patients (34 men and 26 women; mena age = 50 ± 2.2 yr), who served as controls. Antibody to hepatitis C virus was detected with a second-generation enzyme immunoassay using two beads. In chronic hepatitis C patients, no men had thyroid autoantibodies. Nine of 29 women (31 %) had thyroid autoantibodies. Among them, six (20.7%) had high titers of thyroid autoantibodies, and two had hypothyroidism. In all nine of these women, hepatitis C virus viremia was detected on nested polymerase chain reaction (with primers located in the 5′ untranslated region). One year later, titers of thyroid autoantibodies had increased in one patient. Three other patients progressed to hypothyroidism. We judged four of 29 patients (13.8%) to have Hashimoto's thyroiditis on the basis of their high titers of thyroid autoantibodies and biological features of hypothyroidism. In the control group, only one man had thyroid microsome autoantibodies, at a very low titer (1:100). The association between chronic hepatitis C and presence of thyroid autoantibodies is clearly confimed (p = 0.021) by this study. (HEPATOLOGY 1993;18:253-257).
One-step RNA polymerase chain reaction for detection of hepatitis C virus RNA
Authors: Hu Ke-Qin , Yu Chang-Hong , Vierling John M.
Published: March 2007

Although detection of hepatitis C virus RNA with polymerase chain reaction has become the standard for diagnosis, extensive application has been thwarted by polymerase chain reaction's labor intensiveness, risk of false-positive results through contamination and time required for individual assays. To minimize these limitations, we developed and validated a one-step hepatitis C virus RNA polymerase chain reaction assay. The one-step method was compared with traditional hepatitis C virus RNA polymerase chain reaction using primers from the highly conserved 5′ untranslated region of the hepatitis C virus genome. Variables studied in the one-step method included the source and quantity of reverse transcriptase (RTase), the concentration of MgCl2 and the duration of reverse transcription and complementary DNA amplification cycles. Optimal conditions for the one-step method were obtained with 25 U of reverse transcriptase and 2 mmol/L MgCl2. The one-step method substantially reduced the time required for analysis. The sensitivity of the one-step method was comparable to that of traditional hepatitis C virus RNA polymerase chain reaction using serially diluted RNA extracted from the serum of a hepatitis C virus-infected patient. The specificity of the one-step method was confirmed on Southern-blot hybridization. The results exhibited 100% concordance with results of traditional hepatitis C virus RNA polymerase chain reaction in 50 serum samples, including those of positive and negative controls. In addition, 100% concordance was observed between the two methods' results when sera containing low levels of hepatitis C virus RNA were used. In serum samples containing positive- and negative-stranded hepatitis C virus RNA, the one-step method produced stronger polymerase chain reaction product signals than did traditional polymerase chain reaction performed with only an antisense primer for reverse transcription. These results indicated that both strands were reverse-transcribed in the one-step technique. The one-step method is a sensitive and specific alternative to traditional hepatitis C virus RNA polymerase chain reaction. Substantial reductions in labor intensiveness, risks of contamination and time required for analysis make it suitable for testing multiple clinical samples. (Hepatology 1993;18:270-274).
Induced expression of heat-shock protein on biliary epithelium in patients with primary sclerosing cholangitis and primary biliary cirrhosis
Authors: Broomé Ulrika , Scheynius Annika , Hultcrantz Rolf
Published: March 2007

In both primary sclerosing cholangitis and primary biliary cirrhosis it is supposed that immunological mechanisms are involved in the progressive destruction of the bile ducts. The aberrant expression of human leukocyte antigen-DR in the bile ducts of patients with these disorders enables the biliary epithelium to present putative antigens to the surrounding lymphocytes; however, no such antigen has been identified. Heat-shock proteins have been implicated in the pathogenesis of various immunological destructive disorders. Liver biopsy specimens from patients with primary biliary cirrhosis (n = 10) and primary sclerosing cholangitis (n = 13) were compared with those from patients with chronic hepatitis C infection (n = 5) and alcoholic cirrhosis (n = 4) and from normal controls (n = 6). Liver sections were investigated by means of immunohistochemical study using a mouse monoclonal antibody, ML30, directed against the 65-kD heat-shock protein of Mycobacterium, with monoclonal antibody against human leukocyte antigen-DR and with the monoclonal antibody Identi-Tr TCR δ1, which recognizes a determinant on the δ-chain of the γ/δ form of the human T-cell receptor. Human leukocyte antigen-DR expression was found on the biliary epithelium of all primary sclerosing cholangitis and primary biliary cirrhosis patients but not on bile ducts from patients with alcoholic cirrhosis or chronic hepatitis C infection or those from normal controls. The biliary epithelium reacted with ML30 in 9 of 10 primary biliary cirrhosis patients and in all primary sclerosing cholangitis patients. Two different patterns of epithelial staining were observed: a perinuclear pattern and intense staining in the apical part of the cell cytoplasm, as demonstrated with confocal laser scanning microscopy. In the group of patients with chronic hepatitis C infection and alcoholic cirrhosis, ML30 reactivity was less extensive and was present only around the nuclei of bile duct cells. None of the bile ducts in the normal livers reacted with ML30. Staining with ML30 could not be correlated with the histological or clinical staging of the patients. The number and distribution of γ/δ T cells did not differ between the normal liver specimens and the various patient groups. Whether the finding of an induced expression of heat-shock protein in the secretory part of the biliary epithelium in primary biliary cirrhosis and primary sclerosing cholangitis will imply that the heat-shock proteins may act as an immune target recognized by the T cells surrounding the bile ducts in primary biliary cirrhosis and primary sclerosing cholangitis must be elucidated in further studies. (Hepatology 1993;18:298-303).
Serum levels of transforming growth factor-α in patients after partial hepatectomy as determined with an enzyme-linked immunosorbent assay
Authors: Tomiya Tomoaki , Fujiwara Kenji
Published: March 2007

A sandwich enzyme-linked immunosorbent assay for measuring serum transforming growth factor-α levels was developed with monoclonal IgM and polyclonal IgG antihuman transforming growth factor-α antibodies and a system to amplify the activity of the conjugated enzyme. The assay detected serum transforming growth factor-α levels as low as 5 pg/ml. Serum transforming growth factor-α levels were below the detection limit of the assay in 19% of healthy adults, with a mean (± S.D.) detectable level of 22.0 ± 16.7 pg/ml. In 13 patients who underwent partial hepatectomy, serum transforming growth factor-α levels were determined serially after surgery. The levels were increased within 28 days of surgery in all the patients, with a mean maximal level of 118.2 ± 90.8 pg/ml. The maximal level achieved in each case correlated significantly with the resected volume of the liver and the increased volume of the remaining liver 28 days after hepatectomy (r = 0.59, p < 0.05, and r = 0.71, p < 0.005, respectively). In contrast, serum transforming growth factor-α levels showed no increase after laparotomy for nonhepatic surgery. Serum transforming growth factor-α levels may vary depending on the regenerative stimulus and reflect the degree to which liver regeneration will occur in patients after partial hepatectomy. (Hepatology 1993;18:304-308).
Depressed immune function in patients with cirrhosis before emergence of hepatocellular carcinoma
Authors: Saibara Toshiji , Maeda Takashi , Miyazaki Masako , Onishi Saburo , Ymamot Yasutake
Published: March 2007

Hegpatocellular carcinomas 1 cm in diameter with high or low echogenicity can be detected on ultrasonography and confirmed on fine-needle biopsy, but it is still very difficult to detect small hepatocellular carcinomas with isoechogenicity. In this study, we assessed lymphokine-activated killer cell activity and interferon-γ production prospectively every 1 to 3 mo for 23 ± 4 mo (mean ± 1 S.D.) in 227 patients with cirrhosis. Transient depression of lymphokine-activated killer activity was detected in 43 patients (defective lymphokine-activated killer group), and hepatocellular carcinoma was detected in 24 cases before the end of the 18-mo follow-up. Twenty-one (87.5%) of the 24 hepatocellular carcinoma patients were included in the defective lymphokine-activated killer group. Defective lymphokine-activated killer activity was detected more than 6 mo before detection of a space occupying lesion in the liver or elevation of α-fetoprotein level above 400 ng/ml. Serum α-fetoprotein level was elevated above 400 ng/ml in only five cases in which hepatocellular carcinoma was detected as a space-occupying lesion. Our results indicate that sequential assessment of lymphokine-activated killer activity may be a predictor of hepatocellular carcinoma and that the depression of immune function in cirrhotic patients is a serious risk factor for hepatocellular carcinoma emergence. (Hepatology 1993;18:315-319).
Establishment of two distinct human hepatocellular carcinoma cell lines from a single nodule showing clonal dedifferentiation of cancer cells
Authors: Yano Hirohisa , Iemura Akihiro , Fukuda Kazunori , Mizoguchi Atsushi , Haramaki Makoto , Kojiro Masamichi
Published: March 2007

Hepatocellular carcinomas often contain tumor cells of more than one histological grade. The clonal relationship and biological behavior of hepatocellular carcinoma cells in histologically heterogeneous areas have not been fully explored. We established two distinct human hepatocellular carcinoma cell lines (HAK-1A and 1B) from a single nodule showing a three-layered structure with a different histological grade in each layer. Morphologically, HAK-1A and 1B resembled well-differentiated hepatocellular carcinoma cells in the outer layer of the original tumor and poorly differentiated ones in the inner layer, respectively. HAK-1B appeared less differentiated morphologically and more aggressive biologically than HAK-1A; HAK-1B had a shorter doubling time, higher tumorigenicity and an aneuploid DNA index. Chromosome analysis revealed many different abnormalities in the two cell lines, in which, however, two identical structural abnormalities (2q+ and 17p+) were identified. Moreover, sequence analysis of the p53 gene showed identical mutations at codon 242 in both cell lines. These findings suggest that the two cell lines are of clonal origin and that hepatocellular carcinomas consisting of cancerous tissues of more than one histological grade may reflect clonal dedifferentiation in the tumor. Furthermore, we predict that a clonal, morphologically less differentiated subpopulation such as HAK-1B is more aggressive in proliferation and may be closely related to subsequent tumor progression in hepatocellular carcinoma. (Hepatology 1993;18:320-327).
Interferon-γ inhibits liver regeneration by stimulating major histocompatibility complex class II antigen expression by regenerating liver
Authors: Sato Yoshinobu , Tsukada Kazuhiro , Matsumot Yoh , Ab Toru
Published: March 2007

The effects of interferon-γ and interleukin-2 on liver regeneration after 70% hepatectomy in rats was studied immunohistologically, with special attention paid to major histocompatibility complex class II antigen expression. Liver regeneration 2 days after partial hepatectomy as assessed on the basis of bromodeoxyuridine labeling index revealed that regeneration was inhibited significantly in rats given a single dose of interleukin-2 or interferon-γ compared with rats that underwent only partial hepatectomy. Simultaneous administration of interleukin-2 and interferon-γ inhibited liver regeneration more markedly than administration of either drug. In rats subjected to partial hepatectomy, Kupffer cells around the portal vein expressed slightly more major histocompatibility complex class II antigen than did sham-operated controls. In the group given interferon-γ, major histocompatibility complex class II antigen expression was markedly increased. Major histocompatibility complex class II antigen expression was greatest in most Kupffer cells of rats given both interleukin-2 and interferon-γ. These results suggest that interferon-γ activates (proliferating) Kupffer cells, in turn leading to suppression of liver regeneration. These major histocompatibility complex class II antigen-positive Kupffer cells act as antigen-presenting cells and present hepatocyte as antigen, the so-called abnormal self, to helper and cytotoxic T cells. Both types of T cells, in turn, may suppress hepatocyte proliferation. The various cytokines induced by the activated Kupffer cells and helper T cells seem to form a network with interferon-γ to regulate liver regeneration. (Hepatology 1993;18:340-346).
Activin induces cell death in hepatocytes in vivo and in vitro
Authors: Schwall Ralph H. , Robbins Kim , Jardieu Paula , Chang Ling , La Cora , Terrel Timothy G.
Published: March 2007

While studying endocrine responses to activin in female rats, we discovered that activin caused a marked reduction in liver mass. The regressed livers exhibited no gross signs of necrosis or infarction, but histopathological evaluation revealed extensive cell death in the centrilobular regions. The dying cells appeared to fragment into structures resembling apoptotic bodies. Liver mass and histological appearance were restored after cessation of activin infusion, indicating that on an organ level, this effect was reversible. To determine whether the effects observed in vivo were caused by direct actions on the liver, we then tested activin on isolated hepatocytes in serum-free medium. Under these conditions, activin caused many hepatocytes to undergo fragmentation, which was accompanied by a loss of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-reducing activity, an index of viability. We compared the effects of activin with those of transforming growth factor-β because activin is structurally related to transforming growth factor-β and because transforming growth factor-β has been shown previously to induce cell death in hepatocytes. Both proteins caused cell death of comparable magnitude, as defined by the extent of loss of MTT-reducing activity, but transforming growth factor-β was active at one tenth of the effective activin concentrations. A neutralizing monoclonal antibody to transforming growth factor-β blocked the response to transforming growth factor-β but had no effect on the response to activin. Conversely, follistatin, an activinbinding protein, blocked the response to activin but not to transforming growth factor-β. Inhibin, which antagonizes the effects of activin in many systems, had little effect on the response to activin. Activin and transforming growth factor-β differed in their onset of action; exposure to transforming growth factor-β for only 1 hr induced a maximal response, whereas maximal response to activin required its continuous presence for 24 hr. These results show a novel effect of activin on cell death in hepatocytes in vivo and in vitro, suggesting that activin may have a previously unrecognized role in regulating hepatic function. (Hepatology 1993;18:347-356).