bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatitis B virus subtypes and hepatitis C virus genotypes in patients with chronic liver disease in Nepal
Authors: Shrestha Santosh Man , Tsuda Fumio , Okamoto Hiroaki , Tokita Hajime , Horikita Minoru , Tanaka Takeshi , Miyakawa Yuzo , Mayumi Makoto
Published: March 2007

A total of 145 patients with chronic liver disease, including 20 with chronic hepatitis, 63 with cirrhosis and 62 with primary hepatocellular carcinoma from Nepal were tested for markers of hepatitis B virus or hepatitis C virus infection. HBsAg was detected in 57 (39%) and hepatitis C virus RNA in 12 (8%); the cause of liver disease was not known in the remaining 76 (52%). HBsAg was found in 5 (1.3%) of 379 normal controls, whereas hepatitis C virus-associated antibodies were detected in 13 (3.4%), none of whom was positive for serum hepatitis C virus RNA. Subtypes of 102 HBsAg samples, from patients and asymptomatic carriers, were adw in 35 (34%), adr in 4 (4%) and ayw in 48 (47%); the remaining 15 (15%) were of atypical subtypes such as ad, ay and a. Of 12 hepatitis C virus RNA samples, genotype I was detected in 1, genotype II in 5 and genotype V in 1; the remaining five samples were not to be classified by polymerase chain reaction with primers specific for genotypes I to V deduced from hepatitis C virus core sequences, despite high hepatitis C virus RNA titers in all of them. Sequences of 192 amino acids in the entire E1 region of unclassifiable hepatitis C virus isolates from five patients differed from each other in 17% to 23%, and varied from reported isolates of defined genotypes in 13% to 44%. These results indicate that atypical subtypes of hepatitis B virus and novel genotypes of hepatitis C virus would prevail in Nepal. (Hepatology 1994;19:805-809.)
Immunological disorders in C virus chronic active hepatitis: A prospective case-control study
Authors: Pawlotsky Jean-Michel , Yahia Mustapha Ben , Andre Chantal , Voisin Marie-Catherine , Intrator Liliane , Roudot-Thoraval Françoise , Deforges Lionel , Duvoux Christophe , Zafrani Elie-Serge , Duval Jean , Dhumeaux Daniel
Published: March 2007

Hepatitis C virus-related chronic hepatitis may be associated with various immunological disorders. The aim of this study was to determine prospectively the prevalence of the clinical, biochemical and pathological immunological abnormalities in a series of 61 consecutive patients with chronic hepatitis C, compared with those in 61 age- and sex-matched control subjects without markers of hepatitis C virus and hepatitis B virus infections and with those in 61 patients with chronic hepatitis B. The following investigations were systematically performed before any treatment: detection of serum cryoglobulinemia and rheumatoid factor, detection of a large variety of serum antitissue antibodies, biopsy of labial salivary glands, ophthalmological examination, dosage of thyroid-stimulating hormone and in vivo capillary microscopy. Cryoglobulinemia was found in 36% of the hepatitis C virus patients, four of whom had dermatological and/or neurological manifestations of vasculitis, and rheumatoid factor was present in 70%. Serum antitissue antibodies were detected in 41% of cases, mostly antinuclear and anti-smooth muscle antibodies. Liver-kidney microsomal and antithyroid antibodies were rare. Salivary gland lesions were found in 49% of the patients: all had lymphocytic capillaritis, sometimes associated with lymphocytic sialadenitis resembling that of Sjögren’s syndrome, but without features of sicca syndrome and Ro/SSA antibodies. Five percent of the patients had lichen planus. The prevalences of cryoglobulinemia, rheumatoid factor and antitissue antibodies were significantly higher than those in the control group and patients with chronic hepatitis B. In conclusion, hepatitis C virus-related chronic hepatitis may be associated with immunological abnormalities that can be classified into several categories: (a) immune complex-mediated disease, mainly represented by mixed cryoglobulinemia that, in rare cases, may be associated with symptoms of vasculitis; (b) autoimmune abnormalities, frequent and mainly represented by serum autoantibodies, especially antinuclear and anti-smooth muscle antibodies; (c) salivary gland lesions, mainly lymphocytic capillaritis, which is found in approximately half the patients and might secondarily lead to lymphocytic sialadenitis resembling Sjögren’s syndrome; and (d) lichen planus, in which hepatitis C virus could be the main agent of frequently associated liver disease. (Hepatology 1994;19:841-848.)
Strong transient expression of the type I interferon-induced MxA protein in hepatitis A but not in acute hepatitis B and C
Authors: Jakschies Detlef , Zachoval Reinhardt , Müller Rainer , Manns Michael , Nolte Klaus-Ulrich , Hochkeppel Heinz-Kurt , Horisberger Michel-Andre , Deicher Helmuth , Von Wussow Peter
Published: March 2007

The human MxA protein is a new specific marker for type I interferon activity both in vitro and in vivo. In the study presented here this interferon-induced marker, as well as the 2′,5′-oligoadenylate synthetases, was measured in circulating mononuclear cells from 21 patients with acute hepatitis A, 20 patients with acute hepatitis B and 14 patients with acute hepatitis C for determination of the activiation of the interferon system in these viral diseases. In acute hepatitis A a strong expression (10 of 10 patients) of the MxA protein and the 2′,5′-oligoadenylate synthetase activity in peripheral-blood mononuclear cells was observed during the first 2 wk after onset of clinical symptoms. In this period the MxA protein concentrations reached levels similiar to those measured in patients treated with up to 5 × 106 IU interferon-α three times a week. Beyond wk 3, in eight of eight patients with hepatitis A no increased MxA protein levels were found. In contrast, peripheral-blood mononuclear cells from patients with acute hepatitis B contained either no measurable MxA protein or only slightly higher levels of the MxA protein, as did those of most patients (12 of 14) with acute hepatitis C. The MxA protein levels of both hepatitis B and C patients were significantly lower (p < 0.05) than those found in hepatitis A patients. Furthermore, sera from 6 of 10 patients with hepatitis A, but none of 10 patients with acute hepatitis B and C, contained measurable MxA protein. This serum MxA protein may originate from interferon-exposed and subsequently damaged liver cells. These results demonstrate that endogenous interferon is produced in large amounts during the first 2 wk of hepatitis A after onset of symptoms but not thereafter. In contrast, most patients with acute hepatitis B or C produce only minor amounts of interferon but do so for a prolonged period (> 6 wk). We conclude that hepatitis B virus and hepatitis C virus are poor interferon inducers and thus evade an important early defense mechanism against viral infection. (Hepatology 1994;19:857-865.)
Possible multicentric occurrence of hepatocellular carcinoma: A clinicopathological study
Authors: Takenaka Kenji , Adachi Eisuke , Nishizaki Takashi , Hiroshige Kaichiro , Ikeda Toshihiko , Tsuneyoshi Masazumi , Sugimachi Keizo
Published: March 2007

To assess the features of multicentric occurrence in hepatocellular carcinoma, we analyzed 10 of 72 patients (14%) who had undergone hepatic resection for hepatocellular carcinoma from May 1989 to October 1992 both clinically and pathologically. The multicentric occurrence of hepatocellular carcinoma was defined among the simultaneously detected small tumors as (a) at least one tumor consisting of extremely well-differentiated (grade I) hepatocellular carcinoma growing in a replacing pattern or (b) one of a group of hepatocellular carcinomas growing in an area of adenomatous hyperplasia. Of the 10 patients, the tumors in 9 were diagnosed as synchronous multicentric hepatocellular carcinomas, whereas the tumor in 1 was considered metachronous. All patients had cirrhosis; one of them had hepatitis B virus infection and nine patients had HCV infection. The inflammatory findings in the parenchyma were determined on the basis of serum enzyme values (AST, 89 ± 27 IU/L; ALT, 96 ± 43 IU/L). One or two tumors in 9 of 10 patients had thin trabecular or trabecular patterns showing replacing growth. In addition, one of the two tumors in two of nine patients was observed growing in areas of adenomatous hyperplasia. Recurrences were found in 4 of 10 patients. The 3-yr disease-free survival rate was 23%. Multiple recurrences were recognized in the two patients, and in the patients who underwent repeat surgery, grade I tumors were also found. Even though these tumors were small and well-differentiated, the recurrence rate was high. Therefore to detect the recurrence of metachronous multicentric hepatocellular carcinoma at an earlier stage, careful follow-up after surgery should be carried out. (Hepatology 1994;19:889-894.)
Assessment of hepatic iron overload in thalassemic patients by magnetic resonance spectroscopy
Authors: Dixon Ruth M. , Styles Peter , Al-Refaie Faris N. , Kemp Graham J. , Donohue Sarah M. , Wonke Beatrix , Hoffbrand Victor A. , Radda George K. , Rajagopalan Bheeshma
Published: March 2007

The transverse relaxation time of water protons is shortened by the presence of iron. This shortening depends on the amount and the environment of iron in the sample. We have developed a method for measuring short transverse relaxation time noninvasively by magnetic resonance spectroscopy. To evaluate magnetic resonance spectroscopy as a means of assessing hepatic iron content in patients with transfusional iron overload, we compared the results obtained with this method with those obtained by other means of assessing total body iron content. The correlation between the liver biopsy iron concentration and 1/transverse relaxation time was highly significant (r = 0.95, p < 0.004, n = 6) for iron loads up to 3% dry weight. The correlation between serum ferritin and 1/transverse relaxation time was also significant, but the correlation coefficient was much lower (r = 0.67, p < 0.002, n = 20). The correlation between 24-hr urinary iron excretion and 1/transverse relaxation time was not significant, nor was that between AST and 1/transverse relaxation time. We conclude that magnetic resonance spectroscopic determination of the transverse relaxation time of hepatic water is an accurate method of measuring liver iron content, especially when the iron content is below 3%. Because it is a noninvasive method that is associated with negligible side effects, it could provide clinicians with an excellent means of assessing the effectiveness of the various therapeutic strategies used in the management of patients with iron overload. (Hepatology 1994;19:904-910.)
Characteristics of hepatocellular carcinoma originating in the caudate lobe
Authors: Shimada Mitsuo , Matsumata Takashi , Maeda Takashi , Yanaga Katsuhiko , Taketomi Akinobu , Sugimachi Keizo
Published: March 2007

Nine patients with hepatocellular carcinoma originating in the caudate lobe who underwent hepatic resection were studied. The caudate lobe was divided into three parts, according to the criteria of Kumon, including the Spiegel lobe, the paracaval portion and the caudate process. The tumors were located in the Spiegel lobe in four, the paracaval portion in four and the caudate process in one. Surgical procedures consisted of right hepatic lobectomy in one, central bisegmentectomy in one and caudate lobectomy in seven. The mean surgical time was 379 ± 129 min; the mean estimated blood loss was 2,994 ± 2,303 ml. The abovementioned surgical risks were more clearly recognized in the paracaval portion than in the Spiegel lobe. In addition, most patients experienced significant postoperative complications. Six of eight patients with 6 mo or longer follow-up had recurrences, and two of six patients died. The characteristics of hepatocellular carcinoma in the caudate lobe were as follows: (a) a higher surgical risk, and more definite risk in the paracaval portion; and (b) a higher rate of early recurrence. A left lobectomy for the Spiegel lobe, a right or left trisegmentectomy for the paracaval portion and a right lobectomy for the caudate process would be ideal from the point of view of the portal supply of the caudate lobe. However, in cirrhotic patients either a caudate lobectomy or partial resection might be preferable because longer survival can be expected. (Hepatology 1994;19:911-915.)
Murine liver allograft transplantation: Tolerance and donor cell chimerism
Authors: Qian Shigwang , Demetris Anthony J. , Murase Noriko , Rao Abdul S. , Fung John J. , Starzl Thomas E.
Published: March 2007

Nonarterialized orthotopic liver transplantation with no immunosuppression was performed in 13 mouse-strain combinations. Two strain combinations with major histocompatibility complex class I and class II and minor histocompatibility complex disparity had 20% and 33% survival of more than 100 days, but the other 11 combinations, including four that were fully allogeneic and all with only class I, class II or minor disparities, yielded 45% to 100% survival of more than 100 days. Long-living recipients permanently accepted donor-strain heterotopic hearts transplanted on the same day or donor-strain skin 3 mo after liver transplantation, in spite of detectable antidonor in vitro activity with mixed lymphocyte reaction and cellmediated lymphocytotoxicity testing (split tolerance). In further donor-specific experiments, liver grafts were not rejected by presensitized major histocompatibility complex class I-disparate recipients and they protected donor-strain skin grafts from second set (or any) rejection. Less frequently, liver transplantation rescued rejecting skin grafts placed 1 wk earlier in major histocompatibility complex class I, class II and minor histocompatibility complex, class II or minor histocompatibility complex-disparate strain combinations. Donor-derived leukocyte migration to the central lymphoid organs occurred within 1 to 2 hr after liver transplantation in all animals examined, persisted in the surviving animals until they were killed (>375 days), and was demonstrated with double-immunolabeling to be multilineage. The relation of these findings to so-called hepatic tolerogenicity and to tolerance in general is discussed. (Hepatology 1994;19:916-924.)
Inhibition by perhexiline of oxidative phosphorylation and the β-oxidation of fatty acids: Possible role in pseudoalcoholic liver lesions
Authors: Deschamps Dominique , DeBeco Virginie , Fisch Cecile , Fromenty Bernard , Guillouzo Andre , Pessayre Dominique
Published: March 2007

In an attempt to better understand the mechanisms for pseudoalcoholic liver lesions in human beings, we determined the effects of perhexiline on mitochondrial functions in mice and rats. A first series of studies suggested that protonated perhexiline entered mouse mitochondria along the mitochondrial membrane potential. Release of a proton in the mitochondrial matrix led to uncoupling of oxidative phosphorylation, and accumulation of perhexiline inhibited complexes I and II of the respiratory chain, decreased ATP formation in vitro and decreased the mitochondrial β-oxidation of long-, medium- and short-chain fatty acids in vitro and in vivo in mice. In cultured rat hepatocytes, exposure for 24 hr to 25 μmol/L perhexiline markedly decreased hepatocellular ATP and cell viability. Exposure to 5 μmol/L perhexiline did not modify ATP and viability but decreased the β-oxidation of palmitic acid uniformly labeled with carbon 14 by 38%, increased hepatocyte triglyceride levels by 98% and produced microvesicular steatosis after 72 hr of culture. We conclude that perhexiline is concentrated inside mitochondria, where it inhibits both oxidative phosphorylation and the mitochondrial β-oxidation of fatty acids. These effects may contribute to the development of necrosis, steatosis and possibly certain other pseudoalcoholic liver lesions in human beings. (Hepatology 1994;19:948-961.)
Intrasplenic transplantation of isolated periportal and perivenous hepatocytes as a long-term system for study of liver-specific gene expression
Authors: Chen Ling , Davis Gerard J. , Crabb David W. , Lumeng Lawrence
Published: March 2007

Many hepatocyte-specific genes are expressed heterogeneously in the liver lobule depending on the location of the hepatocytes in relation to the inflow or outflow of portal blood (i.e., periportal or perivenous). For example, albumin is expressed in all hepatocytes but more so in the periportal zone, cytochrome P-450IIE1 is exclusively expressed in the perivenous zone and glutamine synthetase is limited to one or two cell layers next to the terminal hepatic venule. Additionally, hepatic damage caused by several xenobiotics, including carbon tetrachloride, is more severe in the perivenous zone. We have isolated highly enriched perivenous and periportal hepatocytes by means of a digitonin-collagenase perfusion method and transplanted them separately into the spleens of syngeneic rats. After transplantation, hepatocyte-specific gene expression in the transplanted perivenous and periportal cells was monitored for up to 13 mo with in situ hybridization to detect the specific gene transcripts (mRNAs). We also studied the effects of carbon tetrachloride administration on transplanted periportal cells by comparing them with intrasplenic transplanted periportal hepatocytes without carbon tetrachloride treatment. Our results showed that: (a) both transplanted perivenous and periportal hepatocytes could survive and proliferate in the splenic microenvironment for a prolonged period; (b) long-term-transplanted periportal hepatocytes in spleen could eventually express a high level of cytochrome P-450IIE1 mRNA in all transplanted hepatocytes and could express glutamine synthetase mRNA in only about 5% to 10% of them, specifically those hepatocytes located adjacent to splenic blood vessels. It is noteworthy that periportal hepatocytes in situ normally do not express the glutamine synthetase gene and express only a low level of cytochrome P-450IIE1 mRNA; and (c) carbon tetrachloride yielded different toxic effects on transplanted periportal hepatocytes at day 3 and mo 8. Necrosis was seen only when transplanted periportal hepatocytes expressed a high level of cytochrome P-450IIE1 mRNA by mo 8. (Hepatology 1994;19:989-998.)
Characterization and enrichment of fetal rat hepatoblasts by immunoadsorption (“panning”) and fluorescence-activated cell sorting
Authors: Sigal Samuel H. , Brill Shlomo , Reid Lola M. , Zvibel Isabel , Gupta Sanjeev , Hixson Douglas , Faris Ronald , Holst Patricia A.
Published: March 2007

We developed methods for enriching fetal hepatoblasts by combining panning and multiparametric fluorescence-activated cell sorting. In unpurified, dissociated fetal liver cell suspensions of embryonic age day 15, 3.2% ± 1.3% and 2.5% ± 0.7% cells expressed albumin and α-fetoprotein, respectively. The remainder exhibited a hemopoietic, endothelial or stromal cell phenotype. Cells were panned first with an antibody to red blood cells to remove erythroid cells and then with monoclonal antibodies OX-43/OX-44 to remove hemopoietic and endothelial cells. This procedure eliminated 84% of fetal hepatic cells, with enrichment of the remainder for albumin or α-fetoprotein expression (up to sixfold increase). Flow cytometric analysis of unlabeled cells revealed two populations, which differed in granularity and autofluorescence. After panning, fluorescence-activated cell sorting for agranular cells yielded OX-43/44-positive cells that were essentially all hemopoietic precursor cells or OX-43/44-negative cells that were mostly hemopoietic precursor cells, along with 3.0% ± 0.7% α-fetoprotein-positive cells. In contrast, sorting for granular, OX-43/44-negative cells enriched for predominantly α-fetoprotein-positive, parenchymal precursor cells (75.1% ± 4.7%). Multiparametric flow cytometric analysis of the expression of an oval cell antigen, OC.3, which is a bile duct and putative liver stem cell marker, showed that most OC.3-positive cells coexpressed OX-43/OX-44 and morphologically were hemopoietic precursor cells. However, approximately 30% of the OX-43/44-negative, granular cells expressed OC.3. Although the physiological significance of OC.3-positive hepatoblasts remains to be determined, the ability to isolate distinct liver cell populations by means of fluorescence-activated cell sorting should facilitate further studies. In addition, because panning alone produced significantly enriched populations of fetal hepatoblasts, applications not requiring further cell purification could be performed with this simple technique. (Hepatology 1994;19:999-1006.)
A new three-dimensional culture system for hepatocytes using reticulated polyurethane
Authors: Sato Yasuo , Ochiya Takahiro , Yasuda Yuko , Matsubara Kenichi
Published: March 2007

Poly-N-para-vinylbenzyl-lactonamide (PVLA)-coated reticulated polyurethane (PVLA-RPU) has been employed for the long-term maintenance of primary rat hepatocyte cultures. After 3 days of incubation of 2 × 107 hepatocytes/cm3 embedded in PVLA-RPU discs and kept in culture medium, most cells showed typical hepatocyte morphology, with some bile canaliculus-like intercellular spaces among the hepatocytes on examination with scanning and transmission electron microscopy. The cells were attached to the surface of the PVLA-RPU and formed multicellular spheroids in the reticulated pores. The hepatocytes maintained various liver-specific functions such as albumin secretion, ammonium metabolism, urea synthesis and gluconeogenesis, and they were viable. The liver-specific functions could be maintained for more than 1 month when the cells were kept in the rats’ peritoneal cavities. This new system may be useful as a bioreactor for an artificial liver. (Hepatology 1994;19:1023-1028.)
Stimulation of phagocytic activity of murine Kupffer cells by tuftsin
Authors: Kubo Shoji , Rodriguez Tom , Roh Mark S. , Oyedeji Caroline , Romsdahl Marvin M. , Nishioka Kenji
Published: March 2007

Tuftsin (Thr-Lys-Pro-Arg) is a natural immunomodulating peptide. We have investigated for the presence of a specific tuftsin receptor on murine Kupffer cells using fluorescein-labeled tuftsin, which retains full biological activity. After incubation with fluorescein-labeled tuftsin, Kupffer cells displayed clear binding of this compound on the plasma membrane. Excess tuftsin inhibited this binding, indicating the presence of specific tuftsin receptors on the Kupffer cells. We then investigated the effect of tuftsin on the phagocytic activity of these cells. Phagocytosis assays were performed on 24-well plates between murine Kupffer cells and fluorescent microspheres. The greatest stimulatory effect of tuftsin on percent phagocytic cells over the control value was observed when the cells were incubated with particles at 1 μg/ml tuftsin for 15 min at 37° C with a particle-to-Kupffer cell ratio of 50:1. Tuftsin also markedly increased the number of particles engulfed by Kupffer cells under the same conditions. These results indicate that Kupffer cells have specific tuftsin receptors; thus tuftsin can stimulate phagocytic activity of Kupffer cells, which constitute the majority of macrophages in the host and are situated strategically in the liver for host defense. (Hepatology 1994;19:1044-1049.)
Of mice, men and cholesterol
Authors: Aftring Paul R. , Freeman Mason W.
Published: March 2007

We employed homologous recombination in embryonic stem cells to produce mice lacking functional LDL receptor genes. Homozygous male and female mice lacking LDL receptors (LDLR−/− mice) were viable and fertile. Total plasma cholesterol levels were twofold higher than those of wild-type littermates, owing to a seven- to ninefold increase in intermediate density lipoproteins (IDL) and LDL without a significant change in HDL. Plasma triglyceride levels were normal. The half-lives for intravenously administered 125I-VLDL and 125I-LDL were prolonged by 30-fold and 2.5-fold, respectively, but the clearance of 125I-HDL was normal in the LDLR−/− mice. Unlike wild-type mice, LDLR−/− mice responded to moderate amounts of dietary cholesterol (0.2% cholesterol/10% coconut oil) with a major increase in the cholesterol content of IDL and LDL particles. The elevated IDL/LDL level of LDLR−/− mice was reduced to normal 4 d after the intravenous injection of a recombinant replication-defective adenovirus encoding the human LDL receptor driven by the cytomegalovirus promoter. The virus restored expression of LDL receptor protein in the liver and increased the clearance of 125I-VLDL. We conclude that the LDL receptor is responsible in part for the low levels of VLDL, IDL, and LDL in wild-type mice and that adenovirus-encoded LDL receptors can acutely reverse the hypercholesterolemic effects of LDL receptor deficiency.
Transarterial oily chemoembolization for the treatment of hepatocellular carcinoma: A multivariate analysis of prognostic factors
Authors: Mondazzi Luca , Bottelli Riccardo , Brambilla Giorgio , Rampoldi Antonio , Rezakovic Indira , Zavaglia Claudio , Alberti Alberto , Idèo Gaetano
Published: March 2007

A total of 84 patients with hepatocellular carcinoma and cirrhosis were analyzed retrospectively to investigate prognostic factors. All patients received transarterial oily chemoembolization as the only anticancer therapy. The follow-up range was 1 to 39 mo (median, 9.5 mo). The overall actuarial survival rates at 12, 24 and 30 mo were 62%, 31% and 24%, respectively. According to univariate analysis, variables significantly associated with survival were age, Child-Pugh grade, total serum bilirubin, Okuda stage, tumor size, degree of labeling of the tumor with Lipiodol, gelatin foam use, changes with treatment in tumor size and changes with treatment in α-fetoprotein concentration. Two multivariate analyses were performed. When pretreatment and treatment variables were considered, parameters with independent prognostic value were age, Child-Pugh grade, total serum bilirubin, tumor size and degree of Lipiodol labeling of the tumor. When follow-up variables were also considered, we (a) confirmed the prognostic significance of all these parameters (age, Child-Pugh grade, total serum bilirubin, tumor size) and (b) found the independent prognostic value of the change in tumor size (or change in α-fetoprotein concentration). Both models yielded different risk coefficients for each class of each variable. Two simple prognostic indexes, based on these coefficients, are proposed: an “initial” index (including pretreatment and treatment variables) and a “follow-up” index (also including follow-up variables). According to the two indexes, the patients were classified into three groups with different prognoses: good (93% and 100% actuarial survival at 1 yr for the initial and follow-up indexes, respectively), intermediate (65% and 53%, respectively) and poor (27% for both indexes). (Hepatology 1994;19:1115-1123.)
Venoocclusive disease of the liver after marrow transplantation: Histological correlates of clinical signs and symptoms
Authors: Shulman Howard M. , Fisher Lloyd B. , Schoch Gary H. , Henne Kevin W. , McDonald George B.
Published: March 2007

We evaluated the relationship between the signs and symptoms of the clinical syndrome called venocclusive disease of the liver after bone marrow transplantation and the histological findings in 76 patients who later came to autopsy. Coded necropsy liver was scored for individual histological features that were correlated with prospectively assessed clinical features that the patients had exhibited during life. Patients were stratified into two groups: those with severe clinical venocclusive disease (n=32) and those without. Clinically severe venocclusive disease was statistically correlated with several zone 3 acinar changes: occluded hepatic venules, the frequency of occluded hepatic venules×degree of occlusion, eccentric luminal narrowing/phlebosclerosis, zone 3 sinusoidal fibrosis and zone 3 hepatocyte necrosis (all p ≤ 0.03). There was a significant relationship between the number of these histological abnormalities in zone 3 of the liver acinus and a clinical diagnosis of severe venocclusive disease (p=0.003). The presence of ascites was significantly correlated with occluded venules, zone 3 sinusoidal fibrosis and zone 3 hepatocyte necrosis (p=0.001). Maximum serum bilirubin in the first 20 days after transplant was significantly correlated with sinusoidal fibrosis, hepatocyte necrosis and eccentric luminal sclerosis/phlebosclerosis (p<0.01) but not with venular occlusion. The clinical syndrome of liver toxicity (commonly called venocclusive disease) that results from cytoreductive therapy before bone marrow transplant is strongly correlated with a constellation of histological lesions involving structures in zone 3 of the liver acinus and the hepatic venules into which sinusoidal blood flows. This study suggests that there is no single diagnostic histological feature. The severity of clinical venocclusive disease appears to be proportional to the number of such histological changes and is not due solely to occlusion of small hepatic venules. (Hepatology 1994;19:1171-1180.)
Hepatic artery and portal vein vascularization of normal and cirrhotic rat liver†
Authors: Kassissia Ibrahim , Brault Antoine , Huet P.-Michel
Published: March 2007

The hepatic artery and portal vein vascularization of seven normal and seven cirrhotic rats was evaluated by means of the multiple-indicator dilution technique using the flow-limited model analysis. Injected 15-μm microspheres were all trapped by the liver in normal and cirrhotic rats after portal vein and hepatic artery injections, ruling out the presence of intrahepatic shunts larger than 15 μm. The albumin curve was linearly displaced relative to the red blood cell curve in both groups of rats, indicating that albumin distribution remained compatible with the flow-limited distribution model. Albumin extravascular space was similar when measured following both routes of injection. Sucrose outflow profile was also compatible with the flow-limited model after portal vein injection in normal rats, but not in severely cirrhotic rats. In contrast, after hepatic artery injection in both normal and cirrhotic rats the sucrose curve was not linearly displaced compared with that of red blood cells; its curve peak was less delayed than its downslope. This finding indicates that, after hepatic artery injection, sucrose distribution was not compatible with the flow-limited model; moreover, its extravascular space was much larger than that after portal vein injection, particularly in cirrhotic rats. This phenomenon is best explained by the peribiliary capillary plexus, lying between terminal arteries and sinusoids, a plexus enlarged in cirrhotic livers. Finally, sinusoidal volume was apparently much larger after hepatic arterial injection compared with that after portal venous injection. This occurrence may well result from an unshared arterial sinusoidal bed or the peribiliary capillary plexus. (Hepatology 1993;18:1189-1197.)
Presinusoidal and proximal intrasinusoidal confluence of hepatic artery and portal vein in rat liver: Functional evidence by orthograde and retrograde bivascular perfusion
Authors: Watanabe Yuji , Püschel Gerhard P. , Gardemann Andreas , Jungermann Kurt
Published: March 2007

The site of confluence of the artery and the portal vein in the liver still appears to be controversial. Anatomical studies suggested a presinusoidal or an intrasinusoidal confluence in the first, second or even final third of the sinusoids. The objective of this investigation was to study the problem with functional biochemical techniques. Rat livers were perfused through the hepatic artery and simultaneously either in the orthograde direction from the portal vein to the hepatic vein or in the retrograde direction from the hepatic vein to the portal vein. Arterial flow was linearly dependent on arterial pressure between 70 cm H2O and 120 cm H2O at a constant portal or hepatovenous pressure of 18 cm H2O. An arterial pressure of 100 cm H2O was required for the maintenance of a homogeneous orthograde perfusion of the whole parenchyma and of a physiologic ratio of arterial to portal flow of about 1:3. Glucagon was infused either through the artery or the portal vein and hepatic vein, respectively, to a submaximally effective “calculated” sinusoidal concentration after mixing of 0.1 nmol/L. During orthograde perfusions, arterial and portal glucagon caused the same increases in glucose output. Yet during retrograde perfusions, hepatovenous glucagon elicited metabolic alterations equal to those in orthograde perfusions, whereas arterial glucagon effected changes strongly reduced to between 10% and 50%. Arterially infused trypan blue was distributed homogeneously in the parenchyma during orthograde perfusions, whereas it reached clearly smaller areas of parenchyma during retrograde perfusions. Finally, arterially applied acridine orange was taken up by all periportal hepatocytes in the proximal half of the acinus during orthograde perfusions but only by a much smaller portion of periportal cells in the proximal third of the acinus during retrograde perfusions. These findings suggest that in rat liver, the hepatic artery and the portal vein mix before and within the first third of the sinusoids, rather than in the middle or even last third. (Hepatology 1994;19:1198-1207.)
Transcriptional rate and steady-state changes of retinoblastoma mRNA in regenerating rat liver†
Authors: Kren Betsy T. , Teel Amy L. , Steer Clifford J.
Published: March 2007

This study characterizes the mRNA expression of the retinoblastoma tumor suppressor gene in regenerating rat liver during 96 hr after 70% partial hepatectomy. A 960-bp Bg1II-OxaNI fragment of murine retinoblastoma cDNA was used to probe Northern blots of poly(A)+-enriched RNA isolated from regenerating liver. Two species of retinoblastoma mRNA, 2.8 kb and 4.7 kb long, were identified in control liver and exhibited an intensity ratio of 5:1, respectively. Expression of the 2.8-kb mRNA was reduced by 50% 1 hr after partial hepatectomy and was less than 10% of control values by 3 hr. The transcript began to reappear at 12 hr and returned to near-baseline levels by 24 hr. In contrast to the rapid disappearance of the 2.8-kb transcript, expression of the 4.7-kb mRNA increased 15-fold by 6 hr and returned to control levels by 18 hr after partial hepatectomy. Pretreatment of the animals with cycloheximide before partial hepatectomy completely stabilized steady-state levels of both mRNA transcripts through 6 hr. Nuclear run-on assays revealed a sixfold increase in transcription by 30 min and a return to near-baseline levels by 6 hr. The in vivo half-lives of the 2.8- and 4.7-kb transcripts in control livers were 39.5 and 41.2 min, respectively. The half-life of the 4.7-kb transcript 6 hr after partial hepatectomy was 39.1 min. Intravenous administration of transforming growth factor-β1, a known inhibitor of hepatocyte replication, just before partial hepatectomy caused no significant change in the modulation of the transcripts through 24 hr. The results indicate that the steady-state expression of the 2.8- and 4.7-kb retinoblastoma mRNA transcripts in the regenerating rat liver is regulated predominately as an intranuclear event. However, in the absence of functional analysis, the significance of these findings in relation to the growth characteristics of the regenerating liver after partial hepatectomy remains unanswered. (Hepatology 1994;19:1214-1222.)
Effects of ethanol feeding on liver, kidney and jejunal membranes of micropigs
Authors: Villanueva Jesús , Chandler Carol J. , Shimasaki Nell , Tang Anna B. , Nakamura Manabu , Phinney Stephen D. , Halsted Charles H.
Published: March 2007

The micropig model of chronic alcoholism was used to study the relationship of lipid composition and physical properties in three different tissue membranes from the same animals. Ethanol feeding reduced membrane anisotropy, as measured with the diphenylhexatriene probe, in liver plasma and kidney brush-border membranes but not in jejunal brushborder membranes. Preincubation with ethanol reduced anisotropy in each of the three control membranes, whereas all three membranes from the ethanol-fed group were relatively tolerant to the acute effect of ethanol. In liver and kidney membranes, ethanol feeding increased levels of linoleic (18:2ω6) acid and decreased levels of arachidonic (20:4ω6) and docosahexaenoic (22:6ω3) acids and their specific double-bond positions, consistent with reduced activities of Δ6 and Δ5 fatty acid desaturases. In liver and kidney membranes, anisotropy parameters and the acute effect of ethanol correlated inversely with levels of linoleic acid and directly with levels of arachidonic and docosahexaenoic acids and their specific double bonds. Levels of docosahexaenoic acid correlated with the acute effect of ethanol in all three membranes. Phospholipid fatty acid profiles were similar in jejunal brush-border membranes and terminal bile samples, suggesting that the effects of ethanol on jejunal fatty acids and physical properties are modulated by intraluminal biliary phospholipids. The effect of ethanol on anisotropy could not be attributed to changes in membrane cholesterol/phospholipid ratios. These studies affirm the value of this new animal model of chronic alcoholism and provide comprehensive evidence for the central role of fatty acid desaturation in the membrane-associated effects of ethanol exposure. (Hepatology 1994;19:1229-1240.)
The immunostimulant OK-432 enhances liver regeneration after 90% hepatectomy in rats
Authors: Kato Kazuya , Matsuda Minoru , Kusano Mitsuo , Onodera Kazuhiko , Kato Junji , Kasai Shinichi , Mito Michio , Hodgson John W.B.
Published: March 2007

The effect of reticuloendothelial system activation on liver regeneration after 90% hepatectomy was investigated. OK-432, a killed streptococcal preparation that increases reticuloendothelial system activity, was administered to rats before 90% partial hepatectomy. Pretreated rats showed marked improvement in long-term survival: 87% (18 of 23) survived beyond 42 hr, compared with only 44.2% (24 of 52) of controls (p<0.05). Survival was determined by means of life-table analysis and regeneration response by means of bromodeoxyuridine labeling index of hepatic DNA. OK-432 pretreatment had significantly increased bromodeoxyuridine labeling index 18, 24, and 42 hr after partial hepatectomy (p<0.05). The results indicate that reticuloendothelial system activation by OK-432 before 90% partial hepatectomy enhances liver regeneration and improves survival, but these factors may not be related. The improved survival may be because of less infection in macrophage-stimulated animals or more rapid clearance of hypotension-causing vasoactive compounds. (Hepatology 1994;19:1241-1244.)
Stimulation of collagen α1(I) gene expression is associated with lipid peroxidation in hepatocellular injury: A link to tissue fibrosis?†
Authors: Bedossa Pierre , Houglum Karl , Trautwein Christian , Holstege Axel , Chojkier Mario
Published: March 2007

We have shown that lipid peroxidation stimulates collagen α1(I) gene transcription in cultured cells. Because increased lipid peroxidation and collagen production coexist in many hepatic disorders, including experimental carbon tetrachloride intoxication, we investigated whether lipid peroxidation modulates collagen gene expression in rats treated with carbon tetrachloride. In this animal model, we show colocalization of increased collagen α1(I) mRNA with lipid peroxidation by means of in situ hybridization and immunohistochemical study for malondialdehyde and 4-hydroxynonenal protein adducts, respectively. However, allyl alcohol treatment, which induced a similar degree of hepatocellular injury but without aldehyde-protein adducts, did not increase collagen α1(I) gene expression, suggesting that hepatocyte necrosis is not sufficient to induce the expression of collagen type I. Furthermore, in the absence of an inflammatory response, coculture experiments of hepatocytes and Ito cells treated with carbon tetrachloride indicate that hepatocytes exert a “paracrine” stimulation of both lipid peroxidation and collagen gene expression in Ito cells. These experiments suggest that hepatocyte lipid peroxidation plays a major role in the regulation of collagen α1(I) gene expression by Ito cells and that it may be a link between hepatocyte injury and hepatic fibrosis. (Hepatology 1994;19:1262-1271.)
Mechanisms of impaired hepatic fatty acid metabolism in rats with long-term bile duct ligation
Authors: Krähenbühl Stephan , Talos Christine , Reichen Jürg
Published: March 2007

Hepatic metabolism of fatty acids is impaired in experimental animals with long-term bile duct ligation. To characterize the underlying defects, fatty acid metabolism was investigated in isolated hepatocytes and isolated liver mitochondria from rats subjected to long-term bile duct ligation or sham surgery. After starvation for 24 hr, the plasma β-hydroxy-butyrate concentration was decreased in rats with bile duct ligation as compared with control rats. Production of β-hydroxybutyrate from butyrate, octanoate and palmitate by hepatocytes isolated from rats subjected to bile duct ligation was also decreased. Liver mitochondria from rats subjected to bile duct ligation showed decreased state 3 oxidation rates for L-glutamate, succinate, duroquinone, and fatty acids but not for ascorbate as substrate. State 3u oxidation rates (uncoupling with dinitrophenol) and activities of mitochondrial oxidascs were also decreased in mitochondria from rats subjected to bile duct ligation. Direct assessment of the activities of the subunits of the electron transport chain revealed reduced activities of complex I, complex II and complex III in mitochondria from rats subjected to bile duct ligation. Activities of the β-oxidation enzymes specific for short-chain fatty acids were all reduced in rats subjected to bile duct ligation. Mitochondrial protein content per hepatocyte was increased by 32% in rats subjected to bile duct ligation compared with control rats. Thus the studies directly demonstrate mitochondrial defects in fatty acid oxidation in rats subjected to bile duct ligation, which explain decreased ketosis during starvation. (Hepatology 1994;19:1272-1281.)
Treatment of chronic hepatitis D virus infection with low and high doses of interferon-α2a: Utility of polymerase chain reaction in monitoring antiviral response
Authors: Madejón Antonio , Cotonat Teresa , Bartolomé Javier , Castillo Inmaculada , Carreño Vicente
Published: March 2007

We examined the efficacy of decreasing high doses (beginning at 18 MU/day) of interferon-α2a vs. that of daily low doses (3 MU) in the treatment of chronic hepatitis delta virus infection. Patients treated with 18 MU had a somewhat higher frequency of normalization of serum ALT levels than patients treated with low doses (31% and 12%, respectively, on an intention-to-treat basis). A decrease in the percentage of hepatitis D virus RNA positivity was observed in both groups at the end of treatment. Thus, whereas in baseline samples 10 (62%) of the patients in each group were positive for hepatitis D virus RNA in serum on slot-blot hybridization, these numbers decreased to 5 (31%) and 4 (25%) patients in groups 1 and 2, respectively, at the end of therapy. However, hepatitis D virus RNA, detected by means of nested polymerase chain reaction, remained in all but two (one in each group) patients who completed the treatment. Finally, during posttreatment follow-up, hepatitis D virus RNA levels returned to baseline values, and only one patient remained negative for this marker. The beneficial effect of inter-feron-a was only transient. Only two patients (one from each treatment group) had persistently normal serum ALT levels after 18 mo of follow-up. Finally, the presence of serum hepatitis D virus RNA at the end of therapy, detected with nested polymerase chain reaction, might be a good marker for the prediction of viral replication relapse. (Hepatology 1994;19:1331-1336.)
Virological and biochemical long-term follow-up of patients with chronic hepatitis c treated with interferon
Authors: Castillo Inmaculada , Bartolomé Javier , Navas Sonia , Gonzalez Sara , Herrero Montserrat , Carreño Vicente
Published: March 2007

We studied the long-term outcomes of 43 patients with chronic hepatitis C treated with one or two interferon cycles, in relation to hepatitis C virus RNA in serum and peripheral-blood mononuclear cells. After the first interferon cycle, 15 (35%) patients had normal transaminase levels, although only five of them had normal levels throughout follow-up (complete responders). After treatment, hepatitis C virus RNA was detected in serum and peripheral-blood mono-nuclear cells with a similar frequency among the five complete responders (60% and 40%, respectively) and the 10 responders with relapse (50% and 20%, respectively). During the follow-up of the complete responders (up to 27 mo), fluctuating viremia levels were found, as demonstrated by the intermittent serum hepatitis C virus RNA positivity. In responders with relapse serum hepatitis C virus RNA reappeared concurrent with the relapse, without changes in peripheral-blood mononuclear cells. A second interferon cycle was performed in 23 nonresponders. Six of them had normalized transaminase levels but four had relapses. After retreatment, hepatitis C virus RNA was detected in peripheral-blood mononuclear cells with the same frequency (50%) in complete responders and in responders with relapse. Loss of serum hepatitis C virus RNA was only achieved in responders with relapse. During the follow-up, half of the complete responders lost serum hepatitis C virus RNA. This marker reappeared in responders with relapse, and hepatitis C virus RNA was found de novo in PBMCs of one responder with relapse. None of the 17 nonresponders to retreatment lost hepatitis C virus RNA in serum or PBMCs during therapy. In the follow-up, serum hepatitis C virus RNA became undetectable in four patients (one of them had normalized transaminase values), whereas viral RNA in peripheral-blood mononuclear cells was detected de novo in seven patients. In summary, hepatitis C virus RNA status in serum or peripheral-blood mononuclear cells after interferon treatment is unrelated to the long-term outcome of the patients. (Hepatology 1994;19:1342-1346.)
Immunohistochemical detection of Fas antigen in liver tissue of patients with chronic hepatitis C
Authors: Hiramatsu Naoki , Hayashi Norio , Katayama Kazuhiro , Mochizuki Kiyoshi , Kawanishi Yuko , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

Apoptosis is a type of cell death that occurs in acute or chronic hepatitis. It has been suggested to be mediated through Fas antigen. To evaluate the role of apoptosis on liver injury of chronic hepatitis C, we studied the expressions of Fas antigen and hepatitis C virus antigen (core antigen) immunohistochemically. Forty liver biopsy samples from patients with type C chronic liver disease were immunostained for Fas antigen and hepatitis C virus antigen. Expression of Fas antigen was found mainly in the cytoplasm of hepatocytes, and these positive cells were found particularly among infiltrating lymphocytes at the advancing edges of “piecemeal necrosis.” The histo-logical activity index showed inflammation of both portal and periportal areas to be more severe in the Fas antigen-positive samples than in the Fas antigen-negative ones (p<0.05 and p<0.001, respectively). Furthermore, semiquantitative analysis revealed more expression of Fas antigen in the liver tissues with active inflammation than in those without it (p<0.01). The prevalence of Fas antigen expression in the hepatitis C virus antigen-positive group was higher than that in the hepatitis C virus antigen-negative group (p<0.05). Our findings suggest that Fas antigen expression (apoptosis) plays an important role in inflammation in the hepatitis C virus-infected liver, particularly in the active inflammation of chronic hepatitis C. (Hepatology 1994; 19:1354-1359.)