bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of the effects of torasemide and furosemide in nonazotemic cirrhotic patients with ascites: A randomized, double-blind study
Authors: Laffi Giacomo , Marra Fabio , Buzzelli Giampiero , Azzena Giuseppe , Meacci Egidio , De Feo Maria Laura , La Villa Giorgio , Gentilini Paolo
Published: March 2007

In a randomized double-blind trial we compared the effects of torasemide, a new loop diuretic, and furosemide in nonazotemic cirrhotic patients with ascites during a 3-day period in association with potassium canrenoate (200 mg/day) administration. Doses of loop diuretics administered in this trial (10 and 25 mg/day of torasemide and furosemide, respectively) had been shown to be equipotent in healthy subjects. Torasemide induced significantly greater natriuresis than furosemide (p <0.02), with a twofold greater percentage increase in basal values (day 1: 130% vs. 50%; day 2: 104% vs. 42%; and day 3: 65% vs. 26%, respectively). Body weight loss was significantly higher during torasemide (p < 0.02) administration, and the overall decrease at the end of the treatment was twice as high for furosemide (2.5 ± 0.6 kg vs. 1.3 ± 0.4 kg, respectively). Diuresis was also higher during torasemide administration, but the difference was not significant (p = 0.08). The extent of kaliuresis observed during the two treatments was almost identical despite the striking differences in the natriuretic response. The effects of the two treatments on plasma electrolytes, creatinine clearance, blood urea nitrogen, mean arterial pressure, heart rate and plasma arginine vasopressin concentration were similar. Both drugs caused increases in plasma renin activity at the end of the treatment, whereas plasma aldosterone concentration slightly increased only after torasemide administration. Despite the presence of a trend toward a more pronounced effect on these parameters after torasemide administration, no significant difference between the two treatments was observed. Blood ammonia increased compared with pretreatment values after furosemide, whereas it remained unmodified after torasemide administration (from 90 ± 15 μg to 112 ± μg and from 87 ± 15 μg to 80 ± 16 μg NH3-N/dl, respectively; p < 0.05 between treatments). The results of this trial indicate that torasemide is suitable for the treatment of sodium and water retention in cirrhotic patients with ascites. Further investigation is needed to assess the long-term tolerability and effectiveness of this new loop diuretic. (Hepatology 1991;13:1101-1105.)
Blood and liver-infiltrating lymphocytes in primary biliary cirrhosis: Increase in activated T and natural killer cells and recruitment of primed memory T cells
Authors: Björkland Anna , Festin Roger , Mendel-Hartvig Ib , Nyberg Anders , Lööf Lars , Tötterman Thomas H.
Published: March 2007

We used two-color and three-color flow cytometric analysis to study phenotypical activation and functional subsets of T and natural killer cells in the blood and liver tissue of patients with primary biliary cirrhosis, other chronic liver diseases and the blood of healthy subjects. The changes in blood lymphocyte phenotype in patients with primary biliary cirrhosis and other chronic liver diseases were similar and comprised elevated relative or absolute numbers of activated human leukocyte antigen-DR+ T subset (CD4+ and CD8+) cells and DR+ natural killer-like (CD16+) cells. B cell (CD19+) numbers were normal. In primary biliary cirrhosis a selective reduction in T cells of suppressor-inducer (CD45RA+ CD4+) type was registered. The human leukocyte antigen-DR expression among CD4 + T cell subsets was investigated further in primary biliary cirrhosis and healthy controls using triple antibody flow cytometric analysis. Phenotypical cell activation was confined to helper T cells of the primed, memory (CD45RO + CD4+) type. The decrease in suppressor-inducer T cells in primary biliary cirrhosis was paralleled by a reciprocal increase in primed memory T cells.

Several significant differences were observed when blood and liver-infiltrating cells from primary biliary cirrhosis patients were compared. In the liver tissue, the CD4/CD8 ratio was decreased, the relative activation of T-subset cells and NK cells was further increased, the suppressor-inducer T subset was further depressed and the primed memory T subset was increased. The cytotoxic T-cell subset (CD11b−) dominated within the CD8+ population. In liver tissue from other chronic liver disease subjects, a lower CD4/CD8 ratio was found compared with primary biliary cirrhosis. However, the infiltrate was relatively similar to primary biliary cirrhosis, indicating a common inflammatory pattern. (Hepatology 1991;13:1106-1111.)
Epidermal growth factor-related transforming growth factors in the urine of patients with hepatocellular carcinoma
Authors: Chuang Lea-Yea , Tsai Juei-Hsiung , Yeh Yun-Chi , Chang Chun-Chang , Yeh Hsing-Wu , Guh Jinn-Yuh , Tsai Jung-Fa
Published: March 2007

To characterize epidermal growth factor-related transforming growth factors in the urine of patients with hepatocellular carcinoma, gel filtration with Bio-Gel P-30 was performed in seven hepatocellular carcinoma patients and seven sex-matched and age-matched healthy controls. Distinct profiles of soft agar growth assay in the hepatocellular carcinoma patients and the normal controls were seen. Three peaks (A, B and C) in the urine were examined. Peak C in most hepatocellular carcinoma patients was higher than that in healthy controls. Similar profiles were detected with epidermal growth factor radioreceptor assay and cellular DNA synthesis assay. This result might indicate that transforming growth factors with low molecular weight were found in the urine of hepatocellular carcinoma patients. An exceptional HCC patient had an additional peak (A′) that corresponded to the high molecular weight protein. We concluded that there were transforming growth factors with functional activity in the urine of patients with hepatocellular carcinoma. (Hepatology 1991;13:1112-1116.)
Regulation of fatty-acid metabolism by pancreatic hormones in cultured human hepatocytes
Authors: Vons Corinne , Pegorier Jean-Paul , Girard Jean , Kohl Claude , Ivanov Marie-Agnès , Franco Dominique
Published: March 2007

The effects of pancreatic hormones and cyclic AMP on long-chain fatty-acid metabolism were investigated in human hepatocytes isolated from 12 liver biopsy specimens and cultured for 4 days in an insulin-free medium. Glucagon (10−6 mol/L) increased endogenous ketone body production by 150%. This resulted from alterations in the partition of long-chain fatty acids from esterification toward oxidation. Glucagon or cyclic AMP enhanced (14C) oleate oxidation (basal = 45.8% ± 5.0%; glucagon = 66.8% ± 5.3%; cyclic AMP = 67.6% ± 5.0% of metabolized oleate) at the expense of oleate esterification. Insulin (10−7 mol/L) antagonized the glucagon-induced oleate oxidation. After 24 hr in basal culture conditions, the rate of lipogenesis decreased to the same low rate as in glucagon-treated cells. The presence of insulin did not restore a high rate of lipogenesis. These results are the first direct evidence of a control of ketone body production by glucagon in the human liver. (Hepatology 1991;13:1126-1130.)
Human fetal hepatocytes respond to inflammatory mediators and excrete bile
Authors: Bauer Joachim , Lengyel Gabriella , Thung Swan N. , Jonas Uwe , Gerok Wolfgang , Acs George
Published: March 2007

Under strict observation of the ethical guidelines of the 1975 Declaration of Helsinki Human Research Committee, primary hepatocyte cultures were prepared from second-trimester fetal liver specimens. We have shown for the first time that fetal hepatocytes have the capacity to produce an acutephase response on treatment with inflammatory mediators. Addition of interleukin-6 to the cultures resulted in strong induction of C-reactive protein and α-1-antichymotrypsin expression, whereas albumin expression was repressed. In contrast to interleukin-6, transforming growth factor-β did not induce C-reactive protein expression. However, as in adult hepatocytes, fetal cells responded to transforming growth factor-β by reduced albumin synthesis. We were able to show by virtue of fluorescein excretion into sealed clefts that fetal hepatocytes have the functional capacity to form bile. Our findings indicate that second-trimester hepatocytes can be regarded as fairly mature liver cells. (Hepatology 1991;13:1131-1141.)
A rat model of acute liver necrosis induced by a monoclonal antibody to liver-specific antigen and complement
Authors: Ikeda Takuya , Kurebayashi Yoichi
Published: March 2007

Acute massive hepatic injury was induced in rats by a monoclonal antibody against a rat liver-specific membrane antigen, and its histological characteristics were investigated. A single intravenous injection of murine ascites containing a monoclonal antibody produced numerous hemorrhagic foci of degenerated and necrotic liver cells predominantly in zones 1 (the periportal area) and 2 (the area of transition between the periportal zone and the perivenular zone) of the liver lobule within 10 min. Massive hepatocellular necroses were observed 1 hr later, but no inflammatory cell infiltration occurred in and around the necrotic foci. Immunohistological study demonstrated marked deposition of the third component of the complement system in the necrotic area. Serum complement activity was sharply decreased immediately after the injection of the antibody, suggesting that the hepatic necrosis is ascribable to a complement-mediated immune attack on the liver cell membrane induced by the antigen-antibody reaction. The hepatic necrosis in response to monoclonal-antibody injection did not progress to a chronic disease and healed almost completely, changing to scar tissues within 2 wk. Although it is not clear whether this hepatic injury has any clinical relevance, this antibody/complement model may be useful for investigating the cause and therapy of hepatic diseases such as fulminant hepatitis. (Hepatology 1991;13:1152-1157.)
Comparative ultrastructural study of rat livers preserved in euro-collins or university of wisconsin solution
Authors: Fratté Serge , Gendrault Jean-Louis , Steffan Anne-Marie , Kirn André
Published: March 2007

University of Wisconsin solution greatly lengthens the time liver storage is possible compared with all previous solutions used. To test whether this improvement is related to better preservation of the endothelial cell, which is thought to be the most vulnerable cell type in cold storage, we compared time-related ultrastructural changes in rat livers stored in this solution or in Euro-Collins solution. Rat livers were harvested after combined arterial and portal perfusion with the cold-storage solution. They were then preserved for different lengths of time in the same solution at 4° C before being perfusion-fixed and processed for light and electron microscopy. The first preservation damage was noted in endothelial cells; the time course of the lesions was similar in both solutions. After 2 hr of storage, enlarged and ruptured fenestrae with many gaps were observed. Swollen at 4 hr, the endothelial cells became stringlike at 10 hr, leading to stripped sinusoidal walls. Hepatocytes appeared better preserved in University of Wisconsin solution. The amount of glycogen, maintained near the control level at 24 hr in the latter, decreased dramatically between 0 and 4 hr in Euro-Collins solution, as ultrastructurally observed and biochemically confirmed. Furthermore, sinusoidal obstruction by blebs originating from the hepatocytes and quantified by image analysis on electron micrographs was markedly delayed. It was significantly less pronounced in University of Wisconsin solution at 24 hr than in Euro-Collins solution at 2 hr (p ≤ 0.05). Our findings confirm that endothelial cells are highly susceptible to preservation damage and show that University of Wisconsin solution does not improve preservation during storage. It is therefore suggested that protection is in part due to a lesser degree of microcirculatory disturbance by blebs stemming from hepatocytes. In addition, endothelial cell structure after preservation does not seem to be a reliable parameter for predicting graft outcome. (Hepatology 1991;13:1173-1180.)
Systemic and splanchnic hemodynamic effects of molsidomine in rats with carbon tetrachloride-induced cirrhosis
Authors: Desmorat Hervé , Vinel Jean-Pierre , Lahlou Omar , Pipy Bernard , Badia Philippe , Cales Paul , Combis Jean-Marc , Souqual Marie-Claude , Pascal Jean-Pierre
Published: March 2007

Molsidomine, a long-acting vasodilator mainly used as an antianginal agent, was reported to decrease the portohepatic venous pressure gradient in patients with alcoholic cirrhosis. This study investigated the effects of linsidomine, the active metabolite of molsidomine, on systemic and splanchnic hemodynamics in rats with CCI4-induced cirrhosis using the microsphere technique. Compared with placebo-treated rats, linsidomine-treated animals were found to have a significant decrease in portal venous pressure (- 18%, p < 0.01) and in mean arterial pressure (-16%, p < 0.01), smaller peripheral resistances (p < 0.01), greater portal venous inflow (p < 0.05), smaller splanchnic arteriolar resistances (p < 0.01) and smaller portocollateral resistances (p < 0.05). Cardiac output, hepatic arterial blood flow, portal blood flow and estimated hepatic blood flow were not significantly different between the two groups of animals. Linsidomine-treated rats exhibited a trend toward greater collateral blood flow compared with controls, but this difference was not significant. We conclude that linsidomine decreases portal venous pressure by reducing portocollateral resistances without affecting liver blood flow. These effects should be beneficial for patients with cirrhosis and portal hypertension. (Hepatology 1991;13:1181-1184.)
Compartmental analysis of steady-state taurocholate transport kinetics by isolated rat hepatocytes
Authors: Coche Thierry , Deroubaix Xavier , Depiereux Eric , Feytmans Ernest
Published: March 2007

We used compartmental modeling to describe taurocholate transport by isolated rat liver cells in suspension. Cells are preincubated in the presence of unlabeled taurocholate. When a steady-state for taurocholate is reached, radiolabeled taurocholate is added to the medium and its exchange kinetics between the medium and the cells are followed over time. Because the studies are performed under steady-state conditions, the kinetics can be described by linear compartmental models. We found a closed two-compartment model sufficient to describe the steady-state transport data. Simulations reveal that if the pools of free and bound intracellular taurocholate exchange rapidly, the cells will behave as a single, kinetically homogeneous compartment and intracellular events will not influence the exchange kinetics of taurocholate between the medium and the cells. The two-compartment model was used to study the concentration dependence of taurocholate transport by isolated cells. Steadystate transport rates and taurocholate concentrations in the medium and the cells were calculated using the model equations. Taurocholate influx, accumulation and efflux processes were studied simultaneously by examining the relationship between appropriate combinations of these variables. Application of this approach to study the inhibition of taurocholate transport by taurochenodeoxycholate is illustrated. In conclusion, this method provides a complementary approach to initial rate studies, which are generally used to investigate bile acid transport by isolated cells. (Hepatology 1991;13:1203-1214.)
Hepatocellular carcinoma in Richardson's ground squirrels (Spermophilus richardsonii): Evidence for association with hepatitis B-like virus infection
Authors: Tennant Bud C. , Mrosovsky N. , McLean Kirsteen , Cote Paul J. , Korba Brent E. , Engle Ronald E. , Gerin John L. , Wright Janet , Michener Gail R. , Uhl Elizabeth , King John M.
Published: March 2007

During studies of seasonal obesity, a high frequency of hepatic neoplasms was observed in Richardson's ground squirrels. Of 12 Richardson's ground squirrels examined thoroughly, 7 had mild or moderate degrees of chronic portal hepatitis and 6 (50%) had hepatocellular carcinoma. Serological tests for hepadnavirus surface antigen, anti-core antibody and virion DNA that recognize the ground squirrel hepatitis virus of California ground squirrels (Spermophilus beecheyi) were uniformly negative. Southern blot analyses of EcoRI digests of liver cell DNA demonstrated 3.2 kb fragments that hybridized with a ground squirrel hepatitis virus-specific probe in nontumorous liver tissue from 6 of 10 ground squirrels and in hepatocellular carcinoma specimens from 2 of 5 squirrels indicating infection with a hepadnavirus related to ground squirrel hepatitis virus. Failure, however, to detect serum antibody to ground squirrel hepatitis core antigen suggested probable antigenic differences between the ground squirrel hepatitis virus of California ground squirrels and the putative Richardson's ground squirrel agent. Further studies are required to fully characterize the hepadnavirus of Richardson's ground squirrels and to determine its relationship to hepatocarcinogenesis in this species. (Hepatology 1991;13:1215-1221.)
Carbohydrate and oxygen metabolism during hepatocellular proliferation: A study in perfused livers from mirex-treated rats
Authors: Yarbrough James , Cunningham Michael , Yamanaka Hirofumi , Thurman Ronald , Badr Mostafa
Published: March 2007

Liver regeneration after partial hepatectomy is accompanied by altered hepatic intermediary metabolism. Because the organochlorine compound mirex also causes liver cell growth, the purpose of this study was to investigate hepatic carbohydrate and oxygen metabolism in perfused livers from mirex-treated rats and to localize cell proliferation in this model. Pretreatment with mirex (100 mg/kg, intragastrically) increased liver/body weight ratios and DNA synthesis in livers of fed rats, effects that were markedly diminished in livers of fasted rats. This finding shows that liver growth caused by mirex, as is the case after partial hepatectomy, is hindered when animals are deprived of food. Furthermore, perfused livers from mirex-treated rats had depleted glycogen stores but significantly elevated oxygen uptake compared with livers from control rats. Increases in oxygen uptake and hepatocellular proliferation were observed mostly in periportal regions of the liver lobule. In regenerating livers, most DNA synthesis was reported to also occur in these regions of the liver lobule. Taken together, these data show that liver cell growth caused by mirex is accompanied by changes in hepatic intermediary metabolism and sublobular proliferation similar to those observed after partial hepatectomy. (Hepatology 1991;13:1229-1234.)
Regenerating nodules-are they premalignant lesions?
Authors: Gumucio Jorge J , Wright Teresa L.
Published: March 2007

In the period 1985-1988, 62 focal liver lesions in 58 cirrhotic patients were studied by ultrasonography; 12 of these focal lesions were documented to be regenerating lesions by echo-guided fine-needle biopsy. During an average follow-up period of 10.2 months (range 3-22 months), hepatocellular carcinoma was subsequently found in 10 of the cases of regenerating nodules, whereas the initial diagnosis of regenerating nodule was confirmed in the remaining two cases. Based upon this finding, it is suggested that every focal mass visualized by ultrasonography in a cirrhotic liver should either considered to be a neoplastic lesion or at least a preneoplastic lesion if the possibility of either a metastatic or benign lesion (eg hemangiomas, focal fatty liver change areas) can be excluded. Therefore either fine-needle aspiration or biopsy of all ultrasonographically revealed mass lesions within a cirrhotic liver is advised, such that early appropriate treatment for hepatocellular carcinoma can be instituted.
Hepatocellular expression of lymphocyte function—associated antigen 3 in chronic hepatitis
Authors: Autschbach Frank , Meuer Stefan C. , Moebius Ulrich , Manns Michael , Hess Georg , Büschenfelde Karl-Hermann Meyer Zum , Thoenes Wolfgang , Dienes Hans-Peter
Published: March 2007

T lymphocyte—mediated cytolytic immune reactions are considered a major cause of hepatocyte injury in chronic viral and autoimmune hepatitis. To further investigate local immune responses, we studied the expression of lymphocyte antigens and cell-cell interaction molecules known to be involved in effector-target cell interactions by light and electron microscopy in liver biopsy specimens from patients with chronic viral and autoimmune hepatitis.

CD8+ lymphocytes were found to be the predominant population of cells in the inflammatory infiltrate in chronic hepatitis B and non-A, non-B hepatitis. In contrast, CD4+ cells constituted a comparably higher proportion of cells and were more numerous than CD8+ cells in chronic autoimmune hepatitis. In both viral and autoimmune hepatitis, a substantial portion of lymphocytes expressed activation antigens such as T11/3 (CD2R) and IL-2-R (CD25). Lymphocyte function—associated antigen-3 (CD58), which mediates lymphocyte adhesion and activation and is the natural ligand of the CD2/T11 lymphocyte surface receptor, could be demonstrated on endothelial cells and hepatocytes. Hepatocellular lymphocyte function—associated antigen-3 expression in chronic hepatitis showed membranous and cytoplasmic staining of hepatocytes and had a positive correlation with the degree of inflammatory activity.

These results suggest that effector-target interactions between hepatocytes and lymphocytes mediated by the lymphocyte function—associated antigen-3/CD2 pathway play a role in chronic inflammatory liver disease. Possible functional consequences of this interaction include enhancement of antigen-specific immune reactions and antigen-independent mechanisms of T cell activation, which may contribute considerably to the degree of inflammatory activity and tissue damage in chronic hepatitis. (HEPATOLOGY 1991;14:223-230.)
Effect of oral propranolol administration on azygos, renal and hepatic uptake and output of catecholamines in cirrhosis
Authors: Bendtsen Flemming , Christensen Niels Juel , Sørensen Thorkild I. A. , Henriksen Jens H.
Published: March 2007

Circulating catecholamines are increased in cirrhosis with portal hypertension, and increase further after propranolol. In 23 cirrhotic patients, plasma norepinephrine and epinephrine were determined in an artery, the azygos vein, the right renal vein and a hepatic vein before and after an oral 80-mg dose of propranolol. Baseline azygos and renal venous norepinephrine levels were significantly higher than arterial norepinephrine levels (+ 20%, p < 0.005; and + 28%, p < 0.001, respectively). Hepatic venous norepinephrine and all venous epinephrine values were below the arterial values (all p < 0.05). After propranolol intake, arterial norepinephrine and epinephrine increased (+ 16%, p < 0.01; and + 93%, p < 0.001, respectively). Significant increases in norepinephrine and epinephrine were found in azygos and renal veins (all p < 0.01), whereas hepatic venous norepinephrine and epinephrine remained unchanged. Azygos and hepatic blood flow decreased after propranolol intake (−27%, p < 0.05; and − 16%, p < 0.01, respectively). Azygos spillover of norepinephrine (an estimate of locally released norepinephrine delivered to the circulation) and clearance of epinephrine remained unaltered. Hepatointestinal clearance showed no significant change for norepinephrine, but showed a borderline-significant decrease for epinephrine (−23%, p = 0.08). Our results show a net production of norepinephrine in the prehepatic splanchnic area drained through superior portalsystemic collaterals and in the kidneys. The increase in circulating catecholamines after propranolol intake is probably due to a combination of further enhancement of sympathetic activity and a decrease in catecholamine degradation. (HEPATOLOGY 1991;14:237-243.)
The metabolic bone disease of primary sclerosing cholangitis
Authors: Hay Eileen J. , Lindor Keith D. , Wiesner Russell H. , Dickson Rolland E. , Krom Ruud A. F. , Larusso Nicholas F.
Published: March 2007

The incidence and severity of osteopenic bone disease in primary sclerosing cholangitis is poorly defined. Clinical, biochemical and radiographic assessment and bone mineral density measurements of the lumbar spine were carried out in two groups of patients. Group 1 consisted of 30 patients with advanced primary sclerosing cholangitis; group 2 consisted of 18 patients with newly diagnosed primary sclerosing cholangitis. Only one patient had bone pain. All patients were normocalcemic; two had elevated serum parathormone levels. Fourteen patients (47%) from group 1 but no patients from group 2 had low serum 25-hydroxyvitamin D levels. Mean bone mineral density was significantly reduced in group 1 patients (0.97 ± 0.04 gm/cm2) compared with age-matched and sex-matched controls (1.25 ± 0.01 gm/cm2, p < 0.0001), and in 15 patients (50%) bone mineral density was below the fracture threshold (0.98 gm/cm2). The bone mineral density in group 2 was not significantly different from controls, and no patient was below the fracture threshold. In neither group did bone mineral density correlate with serum bilirubin, 25-hydroxyvitamin D, fecal fat excretion, previous drug therapy or the presence of chronic ulcerative colitis. Histomorphometrical examination of bone from four group 1 patients showed increased bone resorption, reduced bone formation, moderate-to-severe osteopenia and no osteomalacia. In conclusion, severe osteopenic bone disease is common in advanced primary sclerosing cholangitis and, like that seen in other cholestatis diseases, is consistent with osteoporosis. (HEPATOLOGY 1991;14:257-261.)
Epstein-Barr virus and persistent graft dysfunction after liver transplantation
Authors: Telenti Amadlo , Smith Thomas F. , Ludwig Jurgen , Keating Michael R. , Krom Rund A. F. , Wiesner Russell H.
Published: March 2007

Epstein-Barr virus infection has been associated with a broad spectrum of clinical manifestations, depending on the immune status of the host. In this report, we describe two liver transplant patients who received hepatic allografts from donors serologically positive for Epstein-Barr virus and who experienced primary infection with Epstein-Barr virus associated with prolonged liver graft dysfunction. In both patients, Epstein-Barr serologies converted within 3 mo of liver transplantation, and hepatic histological study revealed mononuclear infiltration of the sinusoids evolving to pronounced immunoblastic features suggestive of evolving lymphoma. In both cases, in situ hybridization studies confirmed the presence of Epstein-Barr virus genome in the liver. Furthermore, polymerase chain reaction analysis suggested that high levels of Epstein-Barr virus DNA were present in biopsy specimens obtained during the episode of acute hepatitis that followed Epstein-Barr virus seroconversion. The degree of Epstein-Barr virus DNA estimated by polymerase chain reaction appeared to increase in parallel with the progression of parenchymal lymphocytic infiltrates. In one patient, a biopsy sample from a cervical node also revealed high levels of Epstein-Barr virus DNA estimated using the polymerase chain reaction technique. Furthermore, in these patients, Epstein-Barr virus DNA levels appeared to decrease dramatically after discontinuing azathioprine administration and beginning treatment with acyclovir. These two cases illustrate the dynamics of Epstein-Barr virus immune regulation and confirm chronic hepatic allograft dysfunction related to Epstein-Barr viral infection. (HEPATOLOGY 1991;14:282-286.)
Binding and internalization of transforming growth factor-β1 by human hepatoma cells: Evidence for receptor recycling
Authors: Sathre Kim A. , Tsang Monica L.-S. , Weatherbee James A. , Steer Clifford J.
Published: March 2007

Cellular processing of 125I-labeled transforming growth factor-β1 was investigated in the human hepatoma cell lines Hep G2 and Hep 3B. Binding of 125I-transforming growth factor-β1 to cell surface receptors was specific, saturable and calciumindependent. Both cell lines exhibited a single class of high-affinity (Kd = 2.2 × 10−10 mol/L) binding sites (4.5 × 103 for the Hep G2 cell; 1.5 × 103 for the Hep 3B cell) for both human and porcine transforming growth factor-β1. Binding was temperature dependent, time dependent and pH dependent. Cell-bound 125I-transforming growth factor-β1 was removed by brief exposure to acidic medium (pH <4) but was converted into an acid-resistant state rapidly after shifting the cells to 37°C. Spontaneous dissociation of bound ligand over a 6 hr period at 4° C was less than 10%. Disuccinimidyl suberate was used to covalently label 125I-transforming growth factor-β1 to cell-surface binding sites. Labeling of the ligand/receptor complexes was inhibited by unlabeled transforming growth factor-β1 but was unaffected by other growth factors. The radiolabeled complexes showed approximate molecular weights of 280,000, 85,000 and 65,000 when run on reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cell-bound 125I-transforming growth factor-β1 was internalized and degraded at 37° C, and the products were released into the medium as trichloroacetic acid-nonprecipitable radioactivity. The lysosomotropic base chloroquine and the carboxylic ionophore monensin inhibited degradation and release of 125I-labeled products from the cells.

In the presence of cycloheximide and under conditions of sustained binding and uptake of saturating amounts of 125I-transforming growth factor-β1 for 3 hr, a 20% decrease in the binding capacity of Hep G2 cells occurred. The result indicates that during active processing of the 125I-transforming growth factor-β1 receptor complex by Hep G2 cells, surface receptors for transforming growth factor-β1 are replenished either from a cryptic intracellular pool or by receptor recycling. (HEPATOLOGY 1991;14:287-295.)
Rates of vertebral bone loss before and after liver transplantation in women with primary biliary cirrhosis
Authors: Eastell Richard , Dickson Rolland E. , Hodgson Stephen F. , Wiesner Russell H. , Porayko Michael K. , Wahner Heinz W. , Cedel Sandra L. , Riggs Lawrence B. , Krom Ruud A. F.
Published: March 2007

Atraumatic fractures caused by osteoporosis may be a serious complication of primary biliary cirrhosis. Mean (± S.D.) bone mineral density in the lumbar spine in 210 ambulatory women with primary biliary cirrhosis was 1.02 ± 0.19 gm/cm2, 7% lower than that in 139 age-matched normal women (after adjustment for age and body weight) (p < 0.001). Bone mineral density in the lumbar spine was inversely related to a risk score index of liver disease severity (r = −0.29, p < 0.001). The mean rate of bone loss in 105 of these 210 women was 2%/yr ± 4%/yr, twice as great as in the 139 normal women (p < 0.02). In 20 women with primary biiary cirrhosis followed up after orthotopic liver transplantation, bone mineral density in the lumbar spine decreased at 3 mo (p < 0.01), and this decrease may have resulted in atraumatic fractures in 13 of them. Bone mineral density in the lumbar spine then increased (p < 0.01) so that by 12 mo the median bone mineral density in the lumbar spine was similar to that before transplantation and by 24 mo it was 5% above it. Therefore we conclude that the progressive bone loss observed in primary biliary cirrhosis (which is further accentuated immediately after transplantation) may be halted, and the bone mass may be restored toward normal within 2 to 3 yr after orthotopic liver transplantation. (HEPATOLOGY 1991;14:296-300.)
In vitro fragmentation of gallstones: Comparison of electrohydraulic, electromagnetic and piezoelectric shockwave lithotripters
Authors: Schneider Thomas H. , Fromm Martin , Ott Roland , Janowitz Paul , Swobodnik Werner , Neuhaus Horst , Ell Christian
Published: March 2007

To compare the fragmentation efficiency of three different shockwave systems, 63 human gallstone triplets were disintegrated in vitro using an electrohydraulic (MPL 9000, Dornier), an electromagnetic (Lithostar Plus, Siemens) and a piezoelectric (Piezolith 2300, R. Wolf) lithotripter. Since each stone triplet was obtained from the same gallbladder, the concrements of one such set were identical in physicochemical parameters. According to the maximal diameter, the calculi were divided into group A (6 to 15 mm) and group B (16 to 30 mm). Shockwave application was terminated when residual fragments measured 4 mm or less. Forty-five triplets were fragmented at energy settings mainly used in clinical treatment of patients with gallbladder stones (MPL 9000: 20 kV; Lithostar Plus: setting 9 (maximal); Piezolith 2300: setting 3). The fragmentation endpoint was achieved in group A (n = 3 × 36) with the Piezolith 2300 after median 150 (range = 50 to 500) pulses and with the Lithostar Plus after 150 (50 to 750) pulses compared with 500 (50 to 1,500) pulses using the MPL 9000 (p < 0.01). In group B (n = 3 × 9) the Lithostar Plus (median = 750, range = 250 to 1,250 pulses) required fewer discharges than the Piezolith 2300 (1,250,250 to 2,500 pulses; p < 0.05) and the MPL (1,500,500 to 1,600 [upper limit] pulses; p < 0.01). At maximal intensities (MPL 9000: 26 kV; Lithostar Plus: setting 9; Piezolith 2300: setting 4), no statistically significant differences between the three tested shockwave systems were observed (Piezolith 2300: 100, 50 to 250 pulses; MPL 9000: 100, 50 to 500 pulses; Lithostar Plus: 100, 50 to 1000 pulses) for group A stones (n = 3 × 12) and group B stones (n = 3 × 6; Piezolith 2300 [500, 250 to 750 pulses], MPL 9000 [625,250 to 750 pulses] and Lithostar Plus [750, 250 to 1,000 pulses]).

An adequate degree of fragmentation independent of the physical shockwave system can be achieved in vitro at maximal intensities for each tested stone size. At submaximal energy settings, better results were obtamed in stones up to 15 mm in diameter with the piezoelectric generator than with the electrohydraulic device, whereas in larger stones disintegration efficiency did not differ significantly between these two lithotripters. (HEPATOLOGY 1991;14:301-305.)
Changes in sulfated proteoglycan production after activation of rat liver macrophages
Authors: Laskin Jeffrey D. , Dokidis Anthea , Gardner Carol R. , Laskin Debra L.
Published: March 2007

Production of extracellular matrix proteins — in particular, the proteoglycans-by macrophages is important in many of their functions, including cell-cell recognition, adhesion and phagocytosis. In this study, we characterized changes in sulfated proteoglycan production by hepatic macrophages following in vivo activation with lipopolysaccharide. We found that both resident Kupffer cells and liver macrophages from lipopolysaccharide-treated rats incorporated [35S]sulfate into proteoglycans. Lipopolysaccharide-activated macrophages incorporated two to three times more of the label than did resident Kupffer cells. In addition, although both cell types produced chondroitin sulfate and heparan sulfate, resident Kupffer cells synthesized more chondroitin sulfate whereas lipopolysaccharideactivated cells produced more heparan sulfate. Using specific antibodies and flow cytometry, we also found that hepatic macrophages produced chondroitin-4-sulfate, chondroitin-6-sulfate and chondroitin-O-sulfate. Lipopolysaccharide-activated macrophages contained more chondroitin-4-sulfate and chondroitin-O-sulfate and less heparan sulfate than did resident Kupffer cells. Both tunicamycin and β-D-xylosides, inhibitors of sulfated proteoglycan biosynthesis, were found to block phagocytosis by the cells. Taken together, these results suggest that sulfated proteoglycans are important in activation and functional responsiveness of liver macrophages. (HEPATOLOGY 1991;14:306-312.)
Bacterial invasion of the biliary system by way of the portal-venous system
Authors: Sung Joseph Y. , Shaffer Eldon A. , Olson Merle E. , Leung Joseph W. C. , Lam Kan , Costerton William J.
Published: March 2007

It has been suggested that bacteria in the intestine gain access into the biliary tract by entering the portal-venous blood. We have tested the hypothesis of hematogenous infection of the biliary system in cats. The animals were treated in three different groups: group A (no biliary obstruction), group B (acute biliary obstruction) and group C (chronic biliary obstruction). A mutant strain of Escherichia coli was infused into the splenic vein of cats at three different dosages (107, 105 and 103) with sham controls. In the unobstructed biliary system, the mutant E. coli was isolated from the bile 30 min and 90 min after the infusion of 107 and 105
E. coli, respectively. No bacteria were found in the bile with the infusion of 103
E. coli and in the control animals. Bile flow was significantly reduced with the infusion of bacteria. The biliary excretion of E. coli in group B was similar to that in group A. In group C, the bile output in the first hour was very high but declined rapidly. E. coli was excreted into the bile at all three dosages of infusion after 30 min. Histological sections of the liver showed that the infused bacteria entered the sinusoidal blood and that some were phagocytosed by Kupffer cells. The portal-venous blood was considered an important route of bacterial invasion into the biliary system, and the penetration of bacteria was facilitated in biliary obstruction. (HEPATOLOGY 1991;14:313-317.)
Liver macrophage-mediated cytotoxicity toward mastocytoma cells involves phagocytosis of tumor targets
Authors: Gardner Carol R. , Wasserman Arthur J. , Laskin Debra L.
Published: March 2007

Macrophage-mediated cytotoxicity toward tumor cells usually involves extracellular lysis of the targets. In this study, we report that liver macrophages from rats treated with lipopolysaccharide (5 mg/kg, intravenous) also kill certain tumor cell targets by phagocytosis. Liver macrophages were coincubated with P815 mouse mastocytoma cells for 24 to 72 hr at an effector/target ratio of 10:1. Macrophage phagocytosis was characterized by flow cytometry and by light and electron microscopy. For flow-cytometric studies, P815 cells were prelabeled with the fluorescent dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethyl-indocarbocyanine perchlorate. We found that coincubation of macrophages with labeled targets resulted in a time dependent increase in macrophage-associated fluorescence, reaching a maximum at 72 hr. This correlated with light-microscopic observations of increased numbers of tumor cells in the macrophages and enhanced macrophage surface area and density. Electron microscopic studies revealed that the initial event in the phagocytic process involved the capture of P815 cells by the pseudopodia of the macrophages. Target cells were then surrounded by lamellipodia, internalized in phagosomes and destroyed. These data, together with previous studies, provide evidence for multiple mechanisms of cytotoxicity mediated by activated liver macrophages. (HEPATOLOGY 1991;14:318-324.)
Functional and morphological features of isolated hepatocytes preserved in university of wisconsin solution
Authors: Sorrentino Dario , Van Ness Karen , Ribeiro Isabela , Miller Charles M.
Published: March 2007

Although several studies suggest that hepatic graft failure after cold ischemia results from nonparenchymal cell damage, other data indicate that hepatocellular ATP content is significantly correlated with the transplantation success rate. In this study, we have conducted a systematic investigation of various aspects of cell viability and function of isolated hepatocytes stored at 4°C for 24 and 48 hr in either University of Wisconsin solution or Hanks’ HEPES buffer, a control solution clinically unsuitable for organ preservation. After 24 hr, hepatocytes stored in Hanks’ HEPES buffer had viability (measured by trypan blue exclusion and ALT and lactic dehydrogenase leakage), transport function (measured by 22Na+ and [3H]taurocholate uptake) and cell size similar or only slightly altered when compared with freshly isolated and University of Wisconsin solution-stored hepatocytes. ATP content was decreased in both groups; however, the reduction was much greater in Hanks’ HEPES buffer-stored cells. Furthermore, ATP regenerating capacity was greatly reduced in Hanks’ HEPES buffer-stored but not in University of Wisconsin solution-stored hepatocytes. By 48 hr viability and function of Hanks’ HEPES buffer-stored hepatocytes were decreased; University of Wisconsin solution afforded partial protection. When examined by light and electron microscopy, cells stored in both University of Wisconsin solution and Hanks’ HEPES buffer for 24 hr appeared essentially normal except for the presence of numerous membrane blebs in the Hanks’ HEPES buffer group. Tissue sections of livers preserved in Hanks’ HEPES buffer but not in University of Wisconsin solution revealed the presence of extensive amounts of blebs in the sinusoidal lumen and loss of endothelial elements. This study indicates that isolated hepatocytes are remarkably resistant to 24-hr cold ischemia. However, the resulting low ATP levels may be responsible for formation of membrane blebs, the shedding of which, in vivo, may lead or contribute to microcirculatory disturbances. These findings may potentially explain the relationship between hepatocellular ATP content and regenerating capacity and the clinical outcome of liver transplantation. (HEPATOLOGY 1991;14:331-339.)
Identification of genetic differences in drug metabolism: Prediction of individual risk of toxicity or cancer
Authors: Nebert Daniel W.
Published: March 2007

These two reports describe recombinant DNA tests that can identify individuals having a defect in the cytochrome P450IID6 (CYP2D6)-mediated oxidative metabolism of debrisoquine and more than two dozen other drugs that are commonly prescribed. The poor metabolizer (PM), representing 5% to 10% of the northern European white population is homozygous for an autosomal recessive trait. Compared with the extensive metabolizer (EM) phenotype, the PM individual is more prone to toxicity caused by some of these drugs. Curiously, the PM phenotype appears to be associated with a lower risk of lung and bladder cancer.

Gough and coworkers propose that the primary gene defect associated with the CYP2D6 polymorphism appears to be a G to A transition mutation affecting the splice junction of intron 3/exon 4 of the CYP2D6 gene. They offer a polymerase chain reaction (PCR) amplification method for diagnosing three variant alleles and claim to have predicted the phenotype of all normal and 34 of 42 (81%) PM individuals. Based on the known frequencies of those variant alleles detectable by this method, however, the assay would be expected to predict less than 60% of PM patients. Heim and Meyer, on the other hand, have combined allelespecific PCR with restriction fragment length polymorphism (RFLP) patterns to identify more than 95% of all mutant alleles, suggesting that their assay would accurately predict more than 90% of all PM individuals. Using this latter assay or a similar test with further improvements, especially in combination with family studies, it should soon be possible for the physician to determine the CYP2D6 phenotype of the patient, thereby avoiding toxic drug overdoses.
Increased expression of intercellular adhesion molecule 1 on bile ducts in primary biliary cirrhosis and primary sclerosing cholangitis
Authors: Adams David H. , Hubscher Stefan G. , Shaw Jean , Johnson Gerald D. , Babbs Christopher , Rothlein Robert , Neuberger James M.
Published: March 2007

It has been suggested that immunological mechanisms involving lymphocyte-mediated damage are important in the characteristic bile-duct damage that occurs in primary biliary cirrhosis and primary sclerosing cholangitis. Because adhesion is necessary for the interaction of lymphocytes with their target structures, we have studied the expression of intercellular adhesion molecule 1, a ligand for the leukocyte adhesion receptor lymphocyte function—associated antigen 1 in the liver of patients with primary biiary cirrhosis and primary sclerosing cholangitis. Strong expression of intercellular adhesion molecule 1 was seen on interlobular bile ducts and proliferating bile ductules in both conditions. In primary biliary cirrhosis, medium-sized ducts, which are spared by the disease, were negative. Minimal bile-duct staining was seen in conditions in which bile-duct damage is not a major feature, such as nonbiliary cirrhosis and acute liver diseases. In patients with cirrhosis from any cause, strong expression of intercellular adhesion molecule 1 was detected on the periseptal hepatocytes adjacent to new connective tissue. The intensity of immunohistochemical staining was recorded using a semiquantitative visual scoring system that was subsequently validated quantitatively by confocal laser scanning microscopy.

The expression/induction of intercellular adhesion molecule 1 on bile ducts may be important in the pathogenesis of bile-duct damage in primary biliary cirrhosis and primary sclerosing cholangitis and is further evidence to support an immune pathogenesis in these two conditions. Furthermore, the induction of intercellular adhesion molecule 1 on hepatocytes may be an important factor in the liver-cell damage and fibrosis that occur during the development of cirrhosis.
Effect of insulin and plasma amino acid concentration on leucine metabolism in cirrhosis
Authors: Petrides Alexander S. , Luzi Livio , Reuben Adrian , Riely Caroline , Defronzo Ralph A.
Published: March 2007

Clinically stable patients with cirrhosis demonstrate insulin resistance with regard to glucose metabolism. However, much less is known about the two major factors, insulin and plasma amino acid concentration, that regulate protein metabolism in cirrhotic patients. To examine this question, we performed paired euglycemic insulin clamp studies in combination with 14Cleucine and indirect calorimetry. In the first study insulin alone was infused, and the plasma amino acid concentration was allowed to decline, During the second study a balanced amino acid solution was infused with insulin to increase the total plasma amino acid concentration approximately twofold. Insulinmediated glucose disposal (4.68 vs. 6.45 mg/kg-min, p < 0.01) was significantly impaired by 30% in cirrhotic patients during both insulin clamp studies. In the postabsorptive state, cirrhotic patients manifested low plasma leucine (76 vs. 102 μmol/L) and α-ketoisocaproate (19 vs. 30 μmol/L) concentrations, but all parameters of leucine turnover were normal. When insulin alone was infused, the endogenous leucine flux (an index of protein degradation) declined similarly in cirrhotic patients (30.8 μmol/m2-min) and control (26.9) subjects, and this was accompanied by a similar decrease in plasma leucine concentration (31% vs. 33%). The decline in circulating leucine concentration was accompanied by a parallel decline in leucine oxidation (5.1 vs. 4.6 μmol/m2-min) and nonoxidative (28.9 vs. 26.0 μmol/m2-min) leucine disposal, which were of similar magnitude in cirrhotic patients and control subjects, respectively. In both cirrhotic patients and control subjects, combined hyperinsulinemia/hyperaminoacidemia elicited a similar stimulation of nonoxidative leucine disposal (an index of protein synthesis) and leucine oxidation while causing a greater suppression of endogenous leucine flux than observed with insulin alone. Thus the suppressive effect of insulin on protein degradation and the stimulatory effect of insulin/amino acid infusion on protein synthesis are not impaired in cirrhotic patients, demonstrating a clear-cut dissociation between the effects of insulin on protein and glucose metabolism.