bluuebunny/crossref_metadata_2025 huggingface dataset:
cyclosporine and liver transplantation: Will the midazolam test make blood level monitoring obsolete?
Authors: Watkins Paul B.
Published: March 2007

Immunosuppression therapy with cyclosporine is often hampered by significant interindividual variability in the metabolic clearance of the drug. It has been suggested that much of the variability in cyclosporine clearance is due to differences in the cytochrome P450 3A4 (CYP3A4) content in the liver and intestinal mucosa. A study was conducted in liver transplant recipients to characterize hepatic CYP3A4 variability during the first 10 days after surgery. The formation of 1′-hydroxymidazolam (1′-OH MDZ) was followed in the plasma after i.v. midazolam (MDZ) administration to 21 multiple-organ donors and to recipients of 10 of the 21 donor livers. Liver biopsy tissue was obtained from donors and recipients after the in vivo pharmacokinetic test. For liver donors, the plasma 1′-OH MDZ/MDZ concentration ratio 30 min after the i.v. MDZ dose was well correlated with the hepatic CYP3A4 content (r = 0.87, P < .001). Much of the variability in the two parameters was attributed to the administration of enzyme-inducing drugs before organ procurement. The mean hepatic CYP3A4 content and plasma 1′-OH MDZ/MDZ concentration ratio in six inducer-treated donors was 4.7-fold and 2.3-fold higher than the respective mean value for all other donors. The hepatic CYP3A4 content and plasma 1′-OH MDZ/MDZ ratio for liver recipients, studied on postoperative day 10, was negatively correlated with the respective parameter measured in donors on day 0 (r = −0.60 for CYP3A4 and r = −0.79 for 1′-OH MDZ/MDZ; P < .05 and P < .01). The dynamic changes in hepatic CYP3A4 expression during the perioperative period, some of which appear to be due to the effect of enzyme-inducing drugs, help explain the difficulties often encountered in the achievement and maintenance of therapeutic cyclosporine blood levels after liver transplantation.
Immunosuppression: Now we have choices
Authors: Sher Linda S. , Makowka Leonard
Published: March 2007

Background.

Tacrolimus (FK 506), a macrolide compound isolated from a bacterium, is a potent immuno-suppressant with activity in solid-organ transplants. Most immunosuppressive regimens for liver transplantation are based on cyclosporine.

Methods.

We conducted an open-label, randomized, multicenter trial to compare the efficacy and safety of tacrolimus-based and cyclosporine-based immunosuppressive regimens for patients receiving a first liver transplant. A total of 478 adults and 51 children (≤12 years of age) were randomly assigned at the time of transplantation to receive tacrolimus (n = 263) or cyclosporine (n = 266) and were followed for one year. The primary end points were patient and graft survival at one year. The secondary end points were the incidence of acute rejection, corticosteroid-resistant rejection, and refractory rejection (continued rejection after two courses of corticosteroids and an intervening course of muromonab-CD3).

Results.

According to Kaplan-Meier analysis, actuarial patient-survival rates at day 360 were 88 percent for both the tacrolimus and cyclosporine groups (P = 0.85; 95 percent confidence interval for the difference, −5.4 to 6.6 percent), and graft-survival rates were 82 percent and 79 percent, respectively (P = 0.55; 95 percent confidence interval for the difference, −4.8 to 9.7 percent). Acute rejection occurred in 154 patients in the tacrolimus group and 173 patients in the cyclosporine group (P <0.002), corticosteroid-resistant rejection occurred in 43 and 82 patients, respectively (P <0.001), and refractory rejection occurred in 6 and 32 patients, respectively (P <0.001). Tacrolimus was associated with a higher incidence of adverse events requiring withdrawal from the study, primarily nephrotoxicity and neurotoxicity; 37 patients in the tacrolimus group and 13 in the cyclosporine group discontinued the study because of adverse events (P <0.001).

Conclusions.

After one year, immunosuppressive regimens based on tacrolimus and cyclosporine were comparable in terms of patient and graft survival. Tacrolimus was associated with significantly fewer episodes of acute, corticosteroid-resistant, or refractory rejection, but substantially more adverse events requiring discontinuation of the drug. (N Engl J Med 1994;331:1110-1115.)
Platelet-derived growth factor receptor expression in hepatic stellate cells: How too much of a good thing can be bad
Authors: Pinzani Massimo
Published: March 2007

A consistent response to liver injury is the activation of resident mesenchymal cells known as lipocytes (Ito, fat-storing cells) into a proliferating cell type. In cultured lipocytes, platelet-derived growth factor (PDGF) is the most potent proliferative cytokine, but requires the activation-dependent expression of its receptor protein (Friedman, S. L., and M. J. P. Arthur. 1989. J. Clin. Invest. 84:1780-1785); the role of PDGF receptor (PDGFR) in liver injury is unknown. We have examined PDGFR gene expression in freshly isolated lipocytes during liver injury and correlated these findings with a culture model of cellular activation. Whereas lipocytes from normal rats had no detectable transcript for the β-PDGFR subunit, this mRNA was induced within 1 h after a dose of carbon tetrachloride (CCI4). In contrast, subunit mRNA was detected in normal cells, but was unchanged after liver injury. Similar results were observed in lipocytes from bile duct-obstructed rats, although β-PDGFR induction was less marked. By immunoblot, induction of β-PDGFR protein in lipocytes isolated from CCI2-treated animals correlated with mRNA increases. In contrast to lipocytes, endothelial cells from normal liver expressed low levels of a- and β-receptor subunit mRNA, which did not increase with injury. Using a P-PDGFR antibody, receptor protein could be identified within fibrotic septa in CCI2-treated animals in regions where cells expressed proliferating cell nuclear antigen (PCNA). In cultured lipocytes activated by growth on uncoated plastic, PPDGFR transcripts appeared within 3 d after plating, which coincided with the onset of cellular proliferation. In contrast, quiescent cells in suspension culture had no detectable β-PDGFR mRNA. These results indicate that β-PDGF receptor induction by lipocytes is an early event during hepatic injury in uiuo and in primary culture. (J. Clin. Invest. 1994; 94:1563-1569.)
Expression of transforming growth factor alpha in the liver before and after interferon alfa therapy for chronic hepatitis B
Authors: Morimitsu Yosuke , Kleiner David E. , Conjeevaram Hari S. , Hsia Chu Chieh , Di Bisceglie Adrian M. , Tabor Edward
Published: March 2007

The effect of interferon alfa (IFN-α) therapy on the expression of transforming growth factor alpha (TGF-α) in the liver during chronic hepatitis B was investigated. Serial liver biopsy specimens were evaluated from 35 patients who had participated in a randomized, controlled trial of recombinant human IFN-α for the treatment of chronic hepatitis B. Percutaneous liver biopsy specimens obtained before and 1 year after entry in the trial were sectioned and stained with a monoclonal antibody to TGF-α in an avidin-biotin-peroxidase-complex system. The expression of TGF-α in each section was evaluated blindly (with respect to treatment group and order of biopsies) and was numerically scored. There was no significant difference in TGF-α expression before or after therapy between 13 patients receiving daily IFN-α, 13 receiving alternate-day IFN-α, and 9 receiving no therapy. Sustained clearance of HBV-DNA and DNA polymerase activity occurred in 8 of 26 treated patients (“responders”); the 18 other patients were “nonresponders.” Expression of TGF-α before IFN-α therapy was significantly higher in responders than in nonresponders; after IFN-α therapy, TGF-α expression decreased significantly among responders compared with nonresponders and untreated controls. Thus, the level of expression of TGF-α in the liver, which was correlated with the severity of inflammation in the liver in this study, appeared to be predictive of the response to IFN-α therapy in chronic hepatitis B, with a higher level of expression indicating a greater likelihood that the patient would respond. (Hepatology 1995; 22:1021-1026.).
Pretreatment serum hepatitis C virus RNA levels and hepatitis C virus genotype are the main and independent prognostic factors of sustained response to interferon alfa therapy in chronic hepatitis C
Authors: Martinot-Peignoux Michele , Marcellin Patrick , Pouteau Mièle , Castelnau Corinne , Boyer Nathalie , Poliquin Marc , Degott Claude , Descombes Isabelle , Le Breton Véronique , Milotova Véronica , Benhamou Jean Pierre , Erlinger Serge
Published: March 2007

The aim of the study was to determine the respective influence of pretreatment serum hepatitis C virus (HCV) RNA levels and HCV genotype on the response to interferon (IFN) alfa in patients with chronic hepatitis C. We retrospectively studied 141 patients with chronic hepatitis C included in two consecutive controlled trials of IFN alfa. A sustained response was observed in 28, a response followed by relapse in 43, and no response in 70 patients. Pretreatment serum HCV RNA quantitation with the branched DNA (bDNA) assay and HCV genotyping with reverse hybridization assay (LiPA) were performed in all patients. Seventy-four percent of the patients had detectable serum HCV RNA (43%, 77% and 84%) in the three groups of patients with sustained response, relapse, and no response, respectively (P = .005). Mean serum HCV RNA level were 1.4 ± 6 × 106, 4.8 ± 6 × 106, and 3.9 ± 5 × 106 genomes/mL in patients with sustained response, response and relapse, and no response, respectively (P < .01). Genotype 1b was found in 7%, 47%, and 46% of the patients in the three response groups, respectively. By univariate analysis, age, source, and duration of HCV infection, serum HCV RNA levels, and HCV genotypes were significantly different in the three response groups. By multivariate analysis, the only independent factors associated with sustained response were low serum HCV RNA levels and HCV genotype other than 1b. Pretreatment serum HCV RNA levels and HCV genotype are the main and independent factors associated with sustained response to IFN therapy. (Hepatology 1995; 22:1050-1056.).
Antipyrine clearance and response to interferon treatment in patients with chronic active hepatitis C
Authors: Coverdale Shirley , Byth Karen , Field Jacqueline , Liddle Christopher , Lin Rita , Farrell Geoffrey C.
Published: March 2007

To determine whether hepatic metabolic function affects the response to interferon treatment, we measured antipyrine clearance (APC) in 85 patients with chronic active hepatitis C and compared the results with treatment outcome. Among 55 patients who responded to interferon by normalization of alanine transaminase (ALT), median APC before treatment was 0.47 (range, 0.12 to 0.98; normal range, 0.34 to 1.02 mL/min/kg body wt), a value that was significantly greater than in 30 nonresponders (0.23; 0.08 to 0.67 mL/min/kg body wt, P < .001). APC was closely associated with response to interferon. The response rate among cases with values >0.25 mL/min/kg body weight was 79%, the same as in cases without cirrhosis. Cases without cirrhosis and with APC of >0.25 mL/min/kg body weight had an 85% chance of responding to interferon; this was unlikely a simple reflection of histological activity, because the correlation with Scheuer score was poor in this subgroup (r = -.31, P < .05). A second, independent group of 43 patients was used to test the predictive value of APC (using 0.25 mL/min/kg body wt as a cut-off) for response to interferon treatment. In this group, APC correctly predicted positive response to interferon in 75% of cases. APC was also used to measure the effects of treatment on hepatic metabolic function. Regardless of outcome, there was no change in APC at the end of a 6-month course of interferon treatment. Six months later, however, improvement in APC (14%; P < .05) was evident among responders but not in those who had failed to respond to interferon. In patients who continued to have normal ALT (18 of 19 tested were also nonviremic), the improvement in APC was sustained for at least 24 months, whereas among relapsers (defined by ALT increase), APC eventually declined to be less than the pretreatment value at 24 months. It is concluded that pretreatment APC is a powerful positive predictor of responsiveness to interferon treatment in patients with chronic hepatitis C, indicating that good hepatic metabolic function may be important in determining the effectiveness of interferon treatment. Furthermore, changes in hepatic metabolic function after apparently successful treatment, at least as determined by APC, appear to be subtle, delayed in onset, and maintained only in those who remain in biochemical remission. (Hepatology 1995; 22:1065-1071.).
Inhibition of woodchuck hepatitis virus replication by adenine arabinoside monophosphate coupled to lactosaminated poly-l-lysine and administered by intramuscular route
Authors: Fiume Luigi , Di Stefano Giuseppina , Busi Corrado , Mattioli Alessandro , Rapicetta Maria , Giuseppetti Roberto , Ciccaglione Anna Rita , Argentini Claudio
Published: March 2007

We prepared a hepatotropic conjugate, suitable for intramuscular (IM) injection, of lactosaminated poly-Llysine with adenine arabinoside monophosphate (ara-AMP), a drug active against hepatitis B virus (HBV). We studied its organ distribution in mice and its antiviral activity in woodchucks that are carriers of woodchuck hepatitis virus (WHV). In mice, after IM administration of a conjugate tritiated in the drug moiety (5.2 μg/g equal to 2 μg/g of ara-AMP) radioactivity in liver was three times greater than in kidney, spleen, and intestine. On the contrary, after IM injection of unconjugated, tritiated, are-AMP (5 μg/g) the amounts of radioactivity in liver, spleen, and kidney were similar. Unconjugated ara-AMP and the conjugate were administered IM to woodchucks for 13 days. Unconjugated ara-AMP decreased viremia at the daily dose of 5 mg/kg but was ineffective at 2.5 mg/kg. The conjugate at the daily doses of 4.2 and 7 mg/kg (equal to 1.5 and 2.5 mg/kg of ara-AMP, respectively) markedly lowered the viremia, which decreased to undetectable levels in the animals treated with the higher dose. Assuming that in HBV-infected patients the same doses will be active, then the amount of conjugate (soluble at 200 mg/mL) required by a 70-kg patient will be contained in a volume of 1.5 to 2.5 mL, compatible with the IM route. Compared with a similar ara-AMP complex with lactosaminated human albumin, currently being studied in clinical trials for the treatment of chronic type B hepatitis, which must be infused intravenously, the present conjugate might provide more patient compliance because of IM administration. (Hepatology 1995; 22:1072-1077.).
Anti-hepatitis B virus activity of a mixture of two monoclonal antibodies in an “inhibition in solution” assay
Authors: Heijtink Rudolf A. , Kruining Johannes , Weber Yvonne A. M. , de Man Robert A. , Schalm Solko W.
Published: March 2007

Two human monoclonal antibodies with anti-hepatitis B activity were investigated separately and as a mixture by means of an “inhibition in solution” assay. With this assay the capacity of anti-HBs antibodies to inhibit the binding of hepatitis B surface antigen (HBsAg) with solid-phase anti-HBs (Ausria II, Abbott Laboratories) was studied. Both HBsAg of different subtypes and purified Dane particles were used. One of the monoclonal antibodies (9H9) was directed against a conformational epitope (anti-“a” like) but induced incomplete inhibition (80% to 90%) of all HBsAg subtypes; the other (4-7B), active against a linear epitope, caused full inhibition of all HBsAg subtypes except one (HBsAg/adw4). In vitro, a mixture of these two monoclonal antibodies was active against HBsAg from liver transplant recipients, including those with suspected variant viruses. (Hepatology 1995; 22:1078-1083.).
Subcellular distribution of large and small hepatitis delta antigen in hepatocytes of hepatitis delta virus superinfected woodchucks
Authors: Cullen John M. , David Caroline , Wang Jia-Gang , Becherer Paul , Lemon Stanley M.
Published: March 2007

Hepatitis delta virus (HDV) encodes only a single protein, the hepatitis delta antigen (HDAg), which is expressed as two molecular forms (large and small) with different functions in viral replication. Compared with small antigen, large antigen has a 19 residue carboxyl terminal extension. Antibodies that recognize a large antigen-specific epitope within this carboxyl extension, or an epitope shared by both large and small antigens (total antigen), were used in immunohistochemical studies of liver sections from superinfected woodchuck carriers of woodchuck hepatitis virus. There were no differences in the subcellular distributions of large and total antigens, with both generally present only in nuclei of hepatocytes. Rare cells demonstrated cytoplasmic staining. Complete or partial granular nucleoplasmic staining with stained nucleoli was the most common pattern observed. Within 31 days of infection, 0.1% to 19% (mean = 7.4%) of all hepatocytes contained antigen. The proportion of these nuclei containing large antigen ranged from 0 to 100% (mean, 39%), and increased during the first month of infection. The number of antigen-positive nuclei and the proportion staining for large antigen were reduced with progression to chronicity, correlating with reductions in the level of viremia. Thus, the large hepatitis delta antigen shares a common subcellular distribution with small antigen and is found in an increasing proportion of the nuclei of infected cells during the course of acute infection. (Hepatology 1995; 22:1090-1100.).
Role of hepatitis C virus in dual and triple hepatitis virus infection
Authors: Liaw Yun-Fan
Published: March 2007

Hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatitis delta virus (HDV) share same transmission routes, thus dual or triple infection may occur and even persist in the same patients. A significant amount of literature has accumulated since the advent of HCV assays. It is pertinent to review and evaluate the clinical and virological significance of HCV in multiple hepatotropic viral infection. The reported series on seroprevalence of HCV indicate that HCV is found in more than 10% of HBV- or HDV-infected patients worldwide. Of the patients with dual or triple infection involving HCV, those having coreplication of viruses tend to have severe and progressive liver disease that is resistant to interferon therapy, in contrast with patients having a single virus infection. Paradoxically, dual or triple hepatitis virus infections are associated with viral interference. In particular, HCV exerts a suppressive effect on HBV and HDV and may enhance seroclearance of HBV antigens or even usurp the role of preexisting virus as the agent for continuing hepatitis. Although HBV and HDV may also suppress HCV, it appears to be less effective. These findings clearly suggest the necessity of monitoring patients with HBV or HDV infections. In view of complex dynamism of viral interaction in multiple hepatotropic virus infection, the importance of HCV assay in the clinical studies can not be overemphasized. The basic mechanisms that regulate the viral interactions, in particular the impact of HCV in dual or triple virus infections, remain to be investigated. (Hepatology 1995; 22:1101-1108.).
The liver-spleen scan as a quantitative liver function test: Correlation with liver severity at peritoneoscopy
Authors: Hoefs John C. , Wang Felix , Kanel Gary , Braunstein Philip
Published: March 2007

Sulfur colloid distribution on liver-spleen scan is determined by the perfused Kupffer cell mass. The perfused Kupffer cell mass is proportional to the perfused hepatocyte mass, but is less affected by acute changes in hepatocyte function. Thus, sulfur colloid distribution parameters (precisely measured by quantitative liver-spleen scan [QLSS]) may be an excellent test of the perfused hepatic mass. Although no gold standard exists for confirmation, a close correlation should exist between liver disease severity assessed at peritoneoscopy and sulfur colloid distribution. Peritoneoscopy severity (scored as total peritoneoscopy score [PS]; range, 0-5) was assessed in 76 patients who also had QLSS. Multivariate equation were generated to estimate liver disease severity from the QLSS. These were then applied prospectively in 20 consecutive patients to validate these equations. In 76 patients, 62 were evaluated because of chronic liver disease (CLD) and included those with micronodular (20) and macronodular (20) cirrhosis with various degrees of severity (Child’s A, 16; B, 29; C, 17). Multivariate analysis yielded a number of combinations of QLSS parameters that correlated with peritoneoscopic severity. These equations were used to estimate liver disease severity. Estimates of liver disease severity (estimated PS [EPS]) correlated well with the PS in these 76 patients (r = .9064; r
2 = .8216; P < .0001). Adding histological fibrosis to the QLSS parameters yields an equation for estimating PS that was even more effective (r = .9462; r
2 = .8953; P < .001). However, validation of multivariate equations requires confirmation of their value in a second population. Applying these equations to a prospective group of 20 patients who subsequently had peritoneoscopic evaluation produced a similar correlation for QLSS parameters alone for estimating severity (r = .870; r
2 = .757; P < .0001), and this was improved when the equation including histological fibrosis was added (r = .936; r
2 = .877; P < .001). We believe these data support the QLSS as a quantitative estimate of the perfused hepatic mass that correlates with liver disease severity at peritoneoscopy. (Hepatology 1995; 22:1113-1121.).
Portal hypertension and iron depletion in patients with genetic hemochromatosis
Authors: Fracanzani Anna Ludovica , Fargion Silvia , Romano Riccardo , Conte Dario , Piperno Alberto , D’Alba Roberta , Mandelli Clara , Fraquelli Mirella , Pacchetti Stefania , Braga Mario , Fiorelli Gemino
Published: March 2007

Clinically, portal hypertension has been considered to be less common and less severe in patients with cirrhosis resulting from iron overload in homozygotes for genetic hemochromatosis than in patients with cirrhosis of other causes. To characterize the prevalence and progression of portal hypertension in genetic hemochromatosis (GH), 120 cirrhosis and iron-overloaded patients were compared with a control group of 120 patients with postnecrotic cirrhosis (PNC) who were matched for gender, age, Child’s class, and alcohol abuse. Gastroesophageal endoscopy and abdominal ultrasonography were performed at diagnosis and repeated every 12 months and every 6 months, respectively, to evaluate the presence and severity of varices, the caliber of the portal vein and its collaterals, and splenic size. At diagnosis a similar frequency of varices was observed in patients with GH (25%) and in PNC (24%), as well as of portal vein abnormalities and spleen enlargement. During the follow-up period, all but two of the patients with GH were treated by phlebotomy and depleted of excess iron. After a mean of 6 ± 4.3 (SD) years of observations (range, 2 to 10 years), varices were improved or completely reversed in 26% of patients with cirrhosis and GH but in only 5% of those with PNC (P < .01). Bleeding from varices was observed in only one patient with GH but in five patients with PNC. Of 22 patients with GH in whom portal hypertension was unmodified or worsened, 16 had coexistent hepatic viral infection. The hazard risk for the development of de novo (i.e., a new occurrence) varices was 17 times higher in patients with PNC than in patients with GH (P < .001). The natural history of portal hypertension in patients with GH was substantially improved by phlebotomy therapy except in the presence of a coexistent hepatic viral infection. These results emphasize the need for intensive phlebotomy therapy even in patients diagnosed after the development of cirrhosis. (Hepatology 1995; 22:1127-1131.).
Reduced serum levels of immunoreactive erythropoietin in patients with cirrhosis and chronic anemia
Authors: Siciliano Massimo , Tomasello Daniele , Milani Alessandro , Ricerca Bianca Maria , Storti Sergio , Rossi Lodovico
Published: March 2007

Chronic anemia is frequently observed in patients affected by cirrhosis. To investigate the possible role of erythropoietin (Epo) in the pathogenesis of anemia in cirrhosis, we measured the immunoreactive Epo levels and the respective hemoglobin (Hb) concentrations in 48 anemic and nonanemic cirrhotic patients and in a control group of healthy subjects and patients with iron-deficiency anemia. Epo concentrations were determined in serum using a sensitive enzyme immunoassay. The regression curve between Epo values and Hb concentrations showed a significant inverse exponential trend both in cirrhotic patients (r = −.55; P < .0001) and controls (r = −.92; P < .0001). In a semilogarithmic plot, the line slope obtained in cirrhotic patients was significantly lower (P < .005) than that of controls, suggesting a blunt Epo response to anemia in cirrhosis. Moreover, covariance analysis showed that the Epo levels for a given degree of anemia were furtherly reduced in the patients with a more severe disease, suggesting a close relation between cirrhosis and the mechanisms involved in the derangement of the Epo feedback system. Finally, the Epo concentrations measured in the cirrhotic patients without anemia did not significantly differ from Epo values obtained in healthy subjects. An impaired Epo response may play a role in maintaining low Hb concentrations in cirrhotic patients with anemia. However, the evidence of a residual Epo response to anemia in cirrhosis and the presence of normal basal Epo levels in nonanemic cirrhotic patients do not support an inadequate Epo secretion as one of the primary causes of anemia in cirrhosis. (Hepatology 1995;22:1132-1135.).
Polymorphisms in alcohol metabolizing enzyme genes and alcoholic cirrhosis in Japanese patients: A multivariate analysis
Authors: Yamauchi Masayoshi , Maezawa Yoshihiko , Mizuhara Yuji , Ohata Mitsuru , Hirakawa Junichi , Nakajima Hisato , Toda Gotaro
Published: March 2007

Alcohol dehydrogenase (ADH), aldehyde dehydrognase (ALDH), and P450IIE1 are the primary enzymes that catalyze the conversion of ethanol to acetaldehyde and then to acetate. Genetic polymorphisms have been reported in ADH2, ADH3, ALDH2, and the 5′-flanking region of P450IIEI. In this study, we used multivariate analysis to determine which genetic polymorphisms in alcohol metabolizing enzymes were independently associated with the development of alcoholic cirrhosis. Thirty-four noncirrhotic alcoholic patients, including 27 with fatty liver and 7 with nonspecific changes, and 46 patients with alcoholic liver cirrhosis were studied. Restriction fragment length polymorphisms (RFLPs) in the ADH2 and P450IIE1 genes were detected by digestion of polymerase chain reaction (PCR)-amplified DNA with MaeIII and RsaI, respectively. In the ALDH2 gene, RFLPs were detected by differences in the MboII site after PCR amplification. By multivariate analysis of four significant factors including total alcohol intake, ADH, ALDH, and P450IIE1 using the multiple logistic regression model, genotype ADH22/ADH22 (P = .029) and genotype c1/c1 of P450IIE1 (P = .013) were found to be independently associated with alcoholic cirrhosis. The odds ratios for ADH22/ADH22 genotype and the type A genotype of P450IIE1 (c1/c1) were 4.600 and 4.006, respectively. These results suggest that ADH2 and P450IIE1 gene polymorphisms may be independently associated with the development of alcoholic liver cirrhosis in Japan. (HEPATOLOGY 1995; 22:1136-1142.).
Differential effects of cytokines on the inducible expression of CYP1A1, CYP1A2, and CYP3A4 in human hepatocytes in primary culture
Authors: Muntané-Relat Jordi , Ourlin Jean-Claude , Domergue Jacques , Maurel Patrick
Published: March 2007

We have investigated the effect of cytokines, including interleukin-6 (Il-6), interleukin-1α (Il-1α), and tumor necrosis factor-alpha (TNF-α), on the inducible expression of cytochrome P450s (CYP) CYP1A1, CYP1A2, and CYP3A4 in human hepatocytes in primary culture. The ability of these cultures to mimic the acute phase response when stimulated with cytokines was evaluated using immunoblotting to measure the production of albumin, ferritin, fibrinogen, and ceruloplasmin. The cytokines exhibited specific patterns of action on the production of these proteins. Albumin was depressed by all the cytokines. In contrast to Il-6 and Il-1α, TNF-α reduced the production of fibrinogen and ceruloplasmin but stimulated the production of ferritin. When cells were treated with the CYP inducer alone, large increases in the expression of CYP1A1 and CYP1A2 by β-naphthoflavone and of CYP3A4 by rifampicin were observed at messenger RNA (mRNA) and protein levels, by ribonuclease protection and immunoblotting, respectively. When the cells were treated with the inducer plus cytokines, the induction of mRNA was greatly reduced. Again, specific patterns of action were revealed: Il-6 had the most potent effect on CYP3A4, whereas TNF-α was the most potent with CYP1A genes. In all cases, changes at the protein levels paralleled changes at the mRNA levels. In cells preinduced with β-naphthoflavone or rifampicin, the decay with time of the levels of the CYP1A2 or CYP3A4 proteins, after the removal of the inducer, was not affected by cytokines. We conclude that cytokines strongly repress the inducibility of CYP1As and CYP3A4 genes at a transcriptional or a posttranscriptional level, but affect neither the rate of translation of CYP mRNAs nor the rate of degradation of the CYP proteins in these cultures. (Hepatology 1995; 22:1143-1153.).
The combination of ursodeoxycholic acid and methotrexate for patients with primary biliary cirrhosis: The results of a pilot study
Authors: Lindor Keith D. , Dickson Rolland E. , Jorgensen Roberta A. , Anderson Monte L. , Wiesner Russell H. , Gores Gregory J. , Lange Stephen M. , Rossi Steven S. , Hofmann Alan F. , Baldus William P.
Published: March 2007

Ursodeoxycholic acid (UDCA) and methotrexate (MTX) have both been proposed as treatments for patients with primary biliary cirrhosis (PBC). It has been suggested that a combination of the two drugs may offer advantages over either used separately. In this pilot study, we sought to evaluate the safety and efficacy of this combination for patients with PBC. Thirty-two patients with antimitochondrial antibody positive PBC were prospectively entered into a pilot study and received UDCA, 13 to 15 mg/kg/d, in conjunction with MTX, 0.25 mg/kg/wk, for a period of 2 years. The results of this treatment were compared with those obtained from 180 patients with PBC studied in a placebo-controlled trial of UDCA alone conducted during the same period. Patients in the pilot study and randomized study were comparable with regard to age, gender, and liver biochemistries. The UDCA/MTX-treated patients were of earlier histologic stage and had a lower mean Mayo risk score. During this period, seven patients in the UDCA/MTX group were withdrawn, four for pulmonary toxicity (two who required hospitalization), and one each with mouth ulcer, extreme fatigue, and hair loss. The use of UDCA/MTX was not associated with improvement in symptoms. In the patients receiving UDCA/MTX, biochemical changes were comparable to those of patients receiving UDCA alone but superior to those in the placebo group (P < .05). Histological changes were comparable in all groups at 2 years. Cessation of MTX while UDCA was continued led to no deterioration in liver biochemistries. In a 2-year study, the use of MTX in combination with UDCA was associated with substantial toxicity, but with no evidence for symptomatic, biochemical, or histologic improvement over that seen with UDCA alone. The use of MTX in patients with PBC should be confined to pa- tients in prospective trials and not used on an empiric basis. (Hepatology 1995; 22:1158-1162.).
Hepatic involvement in mastocytosis: Clinicopathologic correlations in 41 cases
Authors: Mican Joann M. , Di Bisceglie Adrian M. , Fong Tse-Ling , Travis William D. , Kleiner David E. , Baker Bennie , Metcalfe Dean D.
Published: March 2007

Mastocytosis is a disease of mast cell hyperplasia that may involve several organ systems, including liver. Between 1988 and 1991, we conducted a retrospective-prospective study of 41 patients with mastocytosis and found 61% had evidence of liver disease. Hepatomegaly was detected in 24%, splenomegaly in 41%, and elevated serum alkaline phosphatase, serum aminotransaminases, 5′nucleotidase, or gamma-glutamyltranspepti-dase (GGTP) in 54% of the patients. Alkaline phosphatase levels directly correlated with GGTP levels, hepatomegaly, splenomegaly, and liver mast cell infiltration and fibrosis. Elevated alkaline phosphatase levels and splenomegaly were observed more frequently in patients with categories II and III mastocytosis. Five patients in combined disease categories II or III developed ascites or portal hypertension and died of complications of mastocytosis; three had hypoprothrombinemia at the time of death. Thirty-five liver biopsy specimens from 25 patients were examined. Mast cell infiltration was commonly observed in the biopsy specimens, more severe in those patients with either category II or III disease, and correlated with hepatomegaly, splenomegaly, alkaline phosphatase levels, and GGTP levels. Mast cells were often only detected by using special stains (tolu-idine blue and chloracetate esterase). Increased portal fibrosis was seen in 68% of the biopsy specimens and correlated with mast cell infiltration and portal inflammation. Cirrhosis was not observed. Nodular regenerative hyperplasia, portal venopathy, and venoocclusive disease was observed in eight biopsy specimens and may have been the cause of the portal hypertension or ascites in four patients. These findings demonstrate that liver disease with mast cell infiltration is a common finding in patients with mastocytosis. Severe liver disease, however, is uncommon except in patients with mastocytosis who are in categories II and III, where it may contribute to both morbidity and mortality. (Hepatology 1995; 22:1163-1170.).
Ciprofloxacin and long-term prevention of spontaneous bacterial peritonitis: Results of a prospective controlled trial
Authors: Rolachon Alain , Cordier Laurent , Bacq Yannick , Nousbaum Jean-Baptiste , Franza Anne , Paris Jean-Claude , Fratte Serge , Bohn Benoit , Kitmacher Pascal , Stahl Jean-Paul , Zarski Jean-Pierre
Published: March 2007

The aim of this prospective double-blind study was to evaluate the value of long-term antibiotic prophylaxis using ciprofloxacin for the prevention of spontaneous bacterial peritonitis (SBP) in 60 cirrhotic patients with low ascitic fluid protein levels (<15 g/L). The patients were assigned to two groups: group I (n = 28) ciprofloxacin 750 mg per os once a week for 6 months, group II (n = 32) placebo. The two groups were similar for clinical and laboratory characteristics. Twelve patients developed an intercurrent disorder, and 10 patients died during the trial. There were no adverse effects in the treated group. There was a significant decrease in the incidence of SBP (3.6 vs. 22%) (P < .05) and duration of hospitalization (9.3 ± 4.5 vs. 17.6 ± 6.2 days) (P < .05) in the treated group as compared with the placebo group. The bacteriological study showed no acquired resistance to ciprofloxacin after 6 months’ treatment. These results suggest that long-term preventive antibiotic prophylaxis based on the weekly administration of 750 mg of ciprofloxacin is effective in the prevention of SBP in cirrhotic patients. (Hepatology 1995; 22:1171-1174.).
Serum 7α-hydroxycholesterol as a new parameter of liver function in patients with chronic liver diseases
Authors: Kuroki Syoji , Okamoto Shuichiro , Naito Tokio , Oda Hitoshi , Nagase Shoji , Sakai Hironori , Nawata Hajime , Yamashita Hiroyuki , Chijiiwa Kazuo , Tanaka Masao
Published: March 2007

To examine bile acid synthesis in chronic liver diseases, serum total 7α-hydroxycholesterol level was measured by gas-liquid chromatography-mass spectrometry in patients with cirrhosis (n = 23), patients with chronic hepatitis (n = 21), and control subjects (n = 18). The serum 7α-hydroxycholesterol levels were significantly lower in patients with cirrhosis than the controls (78 ± 59 pmol/mL vs. 237 ± 97 pmol/mL; mean ± SD). However, in patients with chronic hepatitis, the level was fully retained (262 ± 102 pmol/mL). Serum 7α-hydroxycholesterol levels of 17 patients with cirrhosis classified as Child B and C ranged from 33 to 69 pmol/mL, and all were less than the normal range (between 104 and 466 pmol/mL), however, those levels of some patients classified as Child A were within the normal range. Serum 7α-hydroxycholesterol levels significantly correlated with serum albumin, cholinesterase, total bile acid, direct bilirubin, alkaline phosphatase, indocyanine green (ICG) retention rate, hepaplastin test, and lecithin-cholesterol acyltransferase activities. We conclude that bile acid synthesis is well preserved in patients with chronic hepatitis and that it is decreased in most patients with cirrhosis. Serum 7α-hydroxycholesterol may be a new parameter of liver function testing to assess hepatic bile acid synthesis in patients with chronic liver diseases. (Hepatology 1995; 22:1182-1187.).
Regulation of bile acid synthesis by deoxycholic acid in the rat: Different effects on cholesterol 7α-hydroxylase and sterol 27-hydroxylase
Authors: Shefer Sarah , Kren Betsy T. , Salen Gerald , Steer Clifford J. , Nguyen Lien B. , Chen Thomas , Tint Stephen G. , Batta Ashok K.
Published: March 2007

We examined the effects of feeding deoxycholic acid (1% and 0.4% of diet), alone and in combination with ursodeoxycholic acid, on serum and biliary bile acid concentrations, hepatic morphology, and the activities and steady-state messenger RNA (mRNA) levels of HMG-CoA reductase and cholesterol 7α-hydroxylase in the rat. Feeding 1% deoxycholic acid increased serum bile acid concentrations (cholestasis), produced portal traid inflammation, bile duct proliferation, and severe hepatocyte necrosis with nuclear pleomorphism. Hepatic demage was preventage when ursodeoxycholic acid (1%) was combined with the deoxycholic acid (1%), or when deoxycholic acid intake was reduced to 0.4%. HMG-CoA reductase and cholesterol 7α-hydroxylase activities were markedly inhibited (−56% and −55%, respectively) with either 1% or 0.4% deoxycholic acid. Ursodeoxycholic acid alone produced an insignificant decline in HMG-CoA reductase and cholesterol 7α-hydroxylase activities, and when combined with 1% deoxycholic acid did not lessen the inhibitory effect of the latter. Steady-state mRNA levels increased 20-fold for HMG-CoA reductase and 53-fold for cholesterol 7α-hydroxylase in rats fed 1% deoxycholic acid. In contrast, 0.4% deoxycholic acid decreased HMG-CoA reductase mRNA levels 76%, and cholesterol 7α-hydroxylase mRNA levels 82%. Ursodeoxycholic acid alone did not affect HMG-CoA reductase or cholesterol 7α-hydroxylase steady-state mRNA levels. Steady-state mRNA levels and activities of sterol 27-hydroxylase, a key enzyme in the alternative acidic pathway of bile acid synthesis, did not change with either high or low doses of deoxycholic acid. In conclusion, 1% deoxycholic acid induced hepatocyte destruction and regeneration associated with increased mRNA levels for HMG-CoA reductase and cholesterol 7α-hydroxylase, but significantly suppressed both enzyme activities. Thus, high-dose deoxycholic acid uncouples HMG-CoA reductase and cholesterol 7α-hydroxylase mRNA levels from enzyme function. In contrast, lower-dose deoxycholic acid (0.4%) inhibited both activities and mRNA levels of HMG-CoA reductase and cholesterol 7α-hydroxylase. Adding 1% ursodeoxycholic acid to 1% deoxycholic acid prevented the rise in mRNA levels but did not lessen the inhibitory effect of the latter. This inhibition occurred without change in hepatic histology, which suggests a regulatory role for deoxycholic acid that is independent of liver damage. Conversely, sterol 27-hydroxylase activity and mRNA levels are not affected by deoxycholic acid treatments. (Hepatology 1995; 22:1215-1221.).
The effect of partial hepatectomy on tumor growth in rats: In vivo and in vitro studies
Authors: de Jong Koert P. , Lont Harold E. , Bijma Amelie M. , Brouwers Mark A. M. , de Vries Elisabeth G. E. , van Veen Marco L. , Marquet Richard L. , Slooff Maarten J. H. , Terpstra Onno T.
Published: March 2007

Residual tumor in the remnant liver after partial hepatectomy (PH) for colorectal liver metastases is a serious clinical problem. This fact is reflected by the high number of recurrences after potentially curative liver resections. Liver regeneration, it appears, might influence the growth of remaining micrometastases in the liver. Using rats, we demonstrated enhancement of growth of a syngeneic colon carcinoma (CC 531) in the remnant liver after 70% PH. Fourteen days after PH, tumor weights in the liver were twice as high as those of sham-operated rats. This difference in tumor weight was not found in extrahepatic tumors. In vitro experiments did not show stimulation of cultured CC 531 cells by portal or systemic serum withdrawn 24 hours or 14 days after hepatectomy as compared with sera obtained after sham operation. Co-cultures of CC 531 cells and hepatocytes (in ratios of 1:10 or 1:1) demonstrated a higher 3H-thymidine incorporation than was the case in separately cultured cells. In co-cultures, bromodeoxyuridine (BrdU) incorporation in DNA was found primarily in CC 531 cells and rarely in hepatocytes. Cell density appeared to be of influence on 3H-thymidine incorporation in co-cultures. Hepatocytes were found to have a stimulating effect on CC 531 cells in low-density cultures, whereas high-density cultures exhibited an inhibiting effect after a culture time of 120 hours. These results show that, depending on cell density in co-cultures, a paracrine stimulating influence of hepatocytes on this type of colon carcinoma cells (CC 531) might be responsible for the increased tumor growth in vivo. (Hepatology 1995; 22:1263-1272.).
Tumor necrosis factor-alpha decreases hepatocyte bile salt uptake and mediates endotoxin-induced cholestasis
Authors: Whiting James F. , Green Richard M. , Rosenbluth Adam B. , Gollan John L.
Published: March 2007

Tumor necrosis factor-alpha (TNF,α), a cytokine that is produced in a variety of inflammatory diseases associated with cholestasis, is believed to be the primary mediator of the systemic effects of endotoxin. Thus, we have investigated the role of TNFα in the pathogenesis of endotoxin-induced cholestasis in intact animals, and in the uptake of taurocholate by cultured hepatocytes. Male Sprague-Dawley rats received either intravenous (IV) endotoxin (7.5 mg/kg) or monoclonal anti-TNFα antibody followed by endotoxin. Basal bile flow and bile salt excretion were measured for a 2-hour period, after which all animals received an IV bolus of taurocholate (10 μmol/100 g body weight). Endotoxin decreased basal bile flow by 41% and bile salt stimulated bile flow by 38% (n = 12;P< .01). Basal bile salt excretion was decreased 86% after endotoxin administration. Passive immunization with anti-TNFα antibody blocked this endotoxin-associated cholestasis. In addition, rat hepatocytes were isolated and cultured in the presence of either endotoxin (10 μg/mL) or TNFα (100 ng/mL) for 24 hours. These primary hepatocyte cultures exhibited a dose- and timedependent, noncompetitive, inhibition of taurocholate uptake. We postulate that TNFα is an important mediator of the cholestasis of sepsis. (Hepatology 1995; 22:1273-1278.).
Introduction of a murine p53 mutation corresponding to human codon 249 into a murine hepatocyte cell line results in growth advantage, but not in transformation
Authors: Dumenco Luba , Oguey Delphine , Wu Justina , Messier Norma , Fausto Nelson
Published: March 2007

The p53 gene is frequently mutated in human tumors; in hepatocellular carcinomas, there is a high frequency of a specific mutation at codon 249 in regions with significant aflatoxin exposure. To assess the role of this p53 mutation in the development of hepatocellular carcinoma, a mutant murine p53 gene, p53ser246, which corresponds to human codon 249, was transfected into a differentiated, nontransformed hepatocyte cell line AML12. Expression of p53ser246 in this line resulted in a growth advantage when compared with either a control vector (which contains a large p53 deletion) or with a different p53 mutant, val135, not found in hepatocellular carcinoma. Overall, there was a threefold increase in colony formation after transfection with p53ser246 as compared with the control or p53val135 vectors, and the p53ser246 plates developed consistently larger colonies. Whereas clones expressing the control or p53val135 constructs showed no significant morphological changes, clones expressing p53ser246 showed increased heterogeneity (large multinucleated cells and areas of small crowded cells) without focus formation. In addition, the ser246 mutation imparted a growth advantage in serum-free media, suggesting less dependence on specific factors present in serum. None of the mutant p53 or control lines were capable of growth in soft agar or tumor formation in nude mice. Thus in this model, in which endogenous wild-type p53 expression is retained, a high level of mutant p53 expression is not sufficient to transform hepatocytes. Our findings indicate that p53ser246 has effects on hepatocytes that may result in a clonal growth advantage and suggest that additional factors are required for the development of hepatocellular carcinoma. (Hepatology 1995; 22:1279-1288.).