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[Fatty acid composition of liver lipids in chronic hepatitis, liver cirrhosis and fatty liver].

Fatty acid spectrum of liver tissue lipids was studied by means of gas-liquid chromatography in 58 bioptic samples from patients with chronic hepatitis, liver cirrhosis and fatty hepatosis. A statistically significant decrease in content of linoleic, arachidonic and docosahexaenic acids as well as an increase in oleic, palmitoleic and myristic acids were found. Under conditions of the diseases studied fatty acid spectrum of liver tissue lipids altered. A decrease in degree of lipid desaturation was especially distinct in liver tissue cirrhosis.

Chromatography, Gas↗

Precursor cells to CD3-intermediate (CD3int) liver mononuclear cells in the adult liver: further evidence for the extrathymic development of CD3int liver mononuclear cells.

Liver mononuclear cells (LMNC) can be divided into CD3-high positive (CD3hi), CD3-intermediate positive (CD3int) and CD3- populations. CD3int LMNC, but not CD3hi LMNC, are considered to be extrathymically derived because they are found in the liver of nude mice and adult thymectomized lethally irradiated bone marrow chimeras. CD3int LMNC express NK1.1 and show a skewed T cell receptor (TCR) V region repertoire with relatively high levels of V beta 8 and V alpha 14 expression, further suggesting that they belong to a different lineage from conventional T cells, which lack NK1.1 expression and V region skewing. Since the liver has been proposed to be a site of extrathymic T cell differentiation in adult mice, we analyzed LMNC for the presence of T cell precursors. CD3- LMNC contained CD4- CD8- cells and CD4lo CD8- cells. Reverse transcription-polymerase chain reaction analysis showed both CD3- populations express the recombination-activating gene -1, which is indispensable in gene rearrangement of TCR and expressed by thymic T cell precursors. Furthermore, when electronically sorted CD3- LMNC were cultured in medium without any feeder cells or exogenously added cytokines for 24 h, CD3int cells, but not CD3hi cells, appeared. These results suggest that the adult liver contains T cell precursors that lack the expression of the CD3/TCR complex, but are strongly committed to differentiate into extrathymic liver CD3int T cells.

Animals↗

[An injury of the liver caused by ischemia-reperfusion in rat liver. Report 2: Relationship between the damage of the liver and during the period of reperfusion].

Liver injury by 30-min ischemia following reperfusion was examined biochemically and histopathologically. A greater increase in the level of LDH was observed after 1-hr reperfusion. However, the level of LDH decreased in proportion to the period of reperfusion, while the levels of GOT and GPT were also increased rapidly and reached its peak at 12 hr following reperfusion and were almost restored to the control level by 48 hr. A similar increase was obtained in the lipid peroxides of the liver. In addition, cyt. P-450 content and NADPH cyt. c reductase activity decreased in proportion to the period of reperfusion up to 12 hr and then recovered by 96 hr. On the other hand, heme oxygenase activity was significantly increased by ischemia-reperfusion. The ischemia-reperfused liver resulted in various morphological changes with the period of reperfusion. The destruction of Disse's space, vacuolization of the cytoplasm and nonviable hepatocytes were observed after 12-hr reperfusion. These results indicate the greatest damages of the liver induced by 30-min ischemia following reperfusion is observed after 12-hr or 24-hr reperfusion. The liver injury by ischemia-reperfusion could be a useful experimental model to develop for future studies.

Alanine Transaminase↗

Activating immunity in the liver. I. Liver dendritic cells (but not hepatocytes) are potent activators of IFN-gamma release by liver NKT cells.

A prominent subset of the hepatic innate immune system is alpha-galactosylceramide (alphaGalCer)-reactive, (CD4(+) and CD4(-)CD8(-)) CD1d-restricted NKT cells. We investigated in C57BL/6 (B6) mice which hepatic cell type stimulates hepatic NKT cell activation. Surface expression of CD1d but not CD40, CD80, or CD86 costimulator molecules was detected in hepatocytes. Pulsed in vitro or in vivo with alphaGalCer, hepatocytes triggered IL-4 release by liver NKT cells but required exogenous IL-12 to trigger IFN-gamma release by NKT cells. Liver dendritic cells (DC) isolated from nontreated mice showed low surface expression of MHC, CD1d, and CD40, CD80, or CD86 costimulator molecules that were strikingly up-regulated after alphaGalCer injection. Although liver CD11c(+) DC displayed lower CD1d surface expression than hepatocytes, they were potent stimulators of IFN-gamma and IL-4 release by liver NKT when pulsed with alphaGalCer in vitro or in vivo. Liver DC are thus potent stimulators of proinflammatory cytokine release by NKT cells, are activated themselves in the process of NKT cell activation, and express an activated phenotype after the NKT cell population is eliminated following alphaGalCer stimulation.

Animals↗

New frontiers in liver surgery. Two-stage liver surgery for the management of advanced metastatic liver disease.

BACKGROUND/AIMS: To assess the value and the safety of main portal branch transection combined with transarterial targeting locoregional neo and adjuvant immunochemotherapy, 32 patients suffering from advanced metastatic liver disease underwent two-stage hepatectomy. METHODOLOGY: From September 1995 to June 1999, 32 consecutive patients underwent two-stage surgery for advanced metastatic liver disease. Firstly we performed ligation and transection of the main portal branch corresponding to the liver lobe occupied by the tumor and introduction of an arterial jet port catheter towards the hepatic artery. After a locoregional transarterial targeting immunochemotherapy regimen the patient had a 2nd laparotomy for hemihepatectomy. Following surgery, locoregional targeting immunochemotherapy was carried out in all patients via the arterial port of the gastroduodenal artery as an adjuvant treatment. RESULTS: There were no operative deaths. Mean survival was 27 +/- 8 months. CONCLUSIONS: Two-stage liver surgery is an appealing alternative that increases the resectability rate and overall survival in patients with advanced metastatic liver disease and is associated with excellent quality of post-operative life.

Adult↗

Liver cell differentiation in a chemically defined medium. Morphologic and enzymatic comparisons of in utero rat liver and liver rudiment grown in organ culture.

The liver diverticulum of the 11-day rat embryo has been grown in a chemically defined medium. Cytodifferentiation and enzymatic activities in the organ culture have been compared with those in the embryonic liver developing in vivo. In the absence of hormone and tissue extracts the liver diverticulum grew and differentiated extensively in the synthetic medium. The appearance and development of cytoplasmic organelles (rough and smooth endoplasmic reticulum, Golgi complex and glycogen particles) and enzymatic activity (glycogen synthetase) in both the embryonic liver and the comparable organ-cultured liver diverticulum were parallel.

Animals↗

Effects of liver failure on branched-chain alpha-keto acid dehydrogenase complex in rat liver and muscle: comparison between acute and chronic liver failure.

BACKGROUND/AIMS: Branched-chain alpha-keto acid dehydrogenase (BCKDH) complex catalyses the committed step in the branched-chain amino acid (BCAA) catabolic pathway. In many cases of liver failure, the serum BCAAs/aromatic amino acids ratio (Fisher's ratio) decreases, and BCAAs have been administered to patients with liver failure to correct this ratio. We conducted an animal study to examine whether the effects on hepatic BCKDH complex differ between acute liver failure (ALF) and chronic liver failure (CLF). METHODS: ALF and CLF was induced in rats by a single high-dose injection and 21 weeks of repeated low-dose injections of carbon tetrachloride, respectively. Plasma BCAA and branched-chain alpha-keto acid (BCKA) levels, and activities and protein amounts of hepatic BCKDH complex and kinase were measured. RESULTS: ALF was characterized by elevated plasma BCAA and BCKA levels and decreased hepatic BCKDH activity. CLF was characterized by decreased plasma BCAA and BCKA levels and increased hepatic BCKDH activity. This increase in BCKDH activity in CLF was associated with the decreased BCKDH kinase, which is responsible for the BCKDH inactivation. CONCLUSIONS: The results obtained in the present study suggest that BCAA catabolism is suppressed in ALF and increased in CLF.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗

Comparison of antigenicity of hepatoma cells, normal liver cells, foetal liver cells and chemically damaged liver cells in guinea-pigs immunized with hepatomata using the macrophage migration inhibition test.

The macrophage migration inhibition test has been used to study the immune responses of guinea-pigs immunized with injections of whole cells of both an allogeneic and a syngeneic hepatoma grown as established cell lines in tissue culture.A clear dose-response relationship between tumour cell concentration and migration inhibition was seen in immunized animals and no significant migration inhibition was seen in control animals. There was no cross reaction between the two tumours used. There was no cross reaction between whole isolated normal liver cells and tumour cells, or between foetal liver cells and tumour cells. Whole isolated liver cells from carbon tetrachloride damaged livers caused some degree of migration inhibition in both normal and immunized guinea-pigs but, taking this into account, they did not appear to cross react with hepatoma cells.

Animals↗

[Liver cirrhosis in autopsy material within 48 years. II. Causes of death, liver cell carcinoma, weights of liver and spleen (author's transl)].

Post mortems of 2289 patients with liver cirrhosis revealed that 37% died of causes related directly to cirrhosis i.e. liver failure or hemorrhage from oesophageal varices. Patients who died of ruptured oesophageal varices in 82% have splenomegaly and most frequently hepatomegaly. Livercell carcinomas were recognized in 7,5% of cirrhotic livers, increasing to 10% in 1976-1978. This rise particularly is conspicuous in women. Liver carcinoma is 4,5 times more frequent in postnecrotic than in portal cirrhosis.

Esophageal and Gastric Varices↗

A liver with polycystic liver disease as graft for orthotopic liver transplantation.

Donor organ shortage calls for widening of criteria for organ transplantation. In exceptional cases, even livers with macroscopic alterations may be accepted for transplantation. The case of a patient who received a liver graft from a donor with previously unknown adult polycystic degeneration is reported. The liver showed only small bilobar cysts with less than 10% of hepatic replacement and was successfully used for orthotopic liver transplantation. Two years after transplantation the patient is in good clinical condition and has returned to work.

Cysts↗

The circulation in liver tissue and experimental liver metastases before and after embolization of the liver artery.

Simultaneous measurement of cardiac output distribution with 86Rubidium and 57Cobalt-tagged microspheres in rats implanted with liver tumors by intraportal injection of sarcoma cells enables quantitation of arterial and portal tumor circulation. The portal circulation was found to be increased in small tumors as compared to the liver, but as the tumor grew there was a decrease in the portal tumor circulation. When the tumor growth became massive even the total liver circulation was reduced, as measured with 133Xenon wash-out. All the tumors had increased arterial circulation. This arterial hyperperfusion was changed into ischemia when the liver artery was occluded through embolization with degradable microspheres.

Animals↗

The appearance and degradation of specific hepatocellular cytoplasmic inclusion bodies in rat liver due to D-galactosamine. I. The relation between the amount of liver glycogen and the appearance of the atypical dense bodies in the liver cell.

One of the most sensitive and specific signs of the galactosamine effect upon the rat liver cell is the appearance of PAS-positive and diastase-resistant granules within the cytoplasm of hepatocytes. Light-microscopic, histochemical, biochemical, and electron-microscopic findings reveal that the appearance of these ADB (= atypical dense bodies) depends upon a working glycogen metabolism at the time of GalN treatment. The ADB are composed of particles resembling, due to shape and size, ribosomes and beta particles of glycogen. Most of them are surrounded by the rER, but they are never enclosed by a limiting membrane. Due to sequential changes they can be generally classified into three types; the early, the intermediate, and the late type. In seven experiments it can be shown, that the appearance of the ADB depends upon the time and dosage after GalN treatment. They occur even if an additional treatment with galactose or uridine prevents the liver from the features of a hepatitis, as also shown in the livers of newborn animals up to 3 weeks of age. The histochemical response against various glucosidases, hexosaminidases, pronase, and RNAse as well as against various fixatives indicates that ADB are composed of, at least, two different constituents, the former RNAse-sensitive and visible with routine light-microscopic staining procedures, the latter RNA-resistant, PAS-positive, and invisible after staining with H & E or toluidine blue. The latter is diastase-resistant, suggesting that this portion of ADB does not represent the usual glycoproteins but some abnormal metabolite of glycogen. The ADB can be detected with maximal accumulation in the cytoplasm of hepatocytes at that time when the glycogen content determined in the liver homogenate by biochemical methods is greatly reduced.

Animals↗

Blood antioxidant levels in patients with alcoholic liver disease correlate with the degree of liver impairment and are not specific to alcoholic liver injury itself.

BACKGROUND: Enhanced production of reactive oxygen species may play a pathogenic role in alcoholic liver injury. AIMS: To investigate whether various antioxidant parameters in blood are affected in different stages of alcoholic liver disease and how specific the changes are relative to non-alcoholic cirrhosis. METHODS: Patients with alcohol abuse without cirrhosis (n=14), with alcoholic cirrhosis [Child-Pugh scores A (n=9), B (n=5) and C (n=18)] and with non-alcoholic cirrhosis [Child-Pugh score C (n=6)] and healthy controls (n=13) were studied. Levels of reduced glutathione and glutathione peroxidase activity in blood, erythrocytic superoxide dismutase activity and carotenoids, alpha-tocopherol and malondialdehyde in plasma were measured. RESULTS: Levels of reduced glutathione were significantly decreased in Child-Pugh score C cirrhotics, alcoholic or not in origin, whereas oxidized glutathione and glutathione peroxidase activity were not affected. Superoxide dismutase activity and alpha-tocopherol levels were not significantly different in the various groups. Carotenoid levels were significantly lower in alcoholic cirrhotics (Child-Pugh score C) vs. controls. Malondialdehyde levels were elevated only in cirrhotics Child-Pugh score C, alcoholic or non-alcoholic. CONCLUSIONS: Levels of reduced glutathione and malondialdehyde reflect the degree of liver impairment, more than the relation with alcohol intake. Decreases in several antioxidant levels are not specific to alcoholic liver injury.

Adult↗

In situ enzymatic oligonucleotide amplification of hepatitis C virus-RNA in liver biopsy specimens (reverse transcriptase in situ polymerase chain reaction) after orthotopic liver transplantation for hepatitis C-related liver disease.

BACKGROUND: Hepatitis C infection recurs after orthotopic liver transplantation for hepatitis C virus (HCV)-related end-stage liver disease. Overlapping histopathologic features may present difficulties in differentiating recurrent HCV in the allograft from other conditions, especially rejection. METHODS: In this study, we evaluated the presence of HCV-RNA by reverse transcriptase in situ polymerase chain reaction (RT in situ RCR) in hepatic tissue, after orthotopic liver transplantation for HCV-related liver disease. Further, detection of HCV-RNA was correlated with the serum HCV-RNA levels, histopathology, and clinical outcome. RESULTS: Twenty-five patients were part of this study. Seventeen patients were transplanted for HCV cirrhosis and eight for an underlying disease other than HCV. None of the patients in the non-HCV group had in situ RT-PCR detection of HCV-RNA. Positive RT in situ PCR for HCV was found in 9 of 17 HCV patients, and the patients had a clinical course consistent with recurrent hepatitis C disease. Four of these nine patients had an initial histologic diagnosis of rejection. The other eight patients in the HCV group had negative RT in situ PCR, and none of them had a course compatible with recurrent HCV disease, although four patients were histologically diagnosed as having chronic C hepatitis. The mean HCV-RNA level (log/mL) in the patients who had in situ detection of HCV-RNA was 7.01+/-0.26. Although RT-PCR was negative in 8 of 17 HCV patients, the patients were serologically viremic and the mean HCV-RNA level was 6.05+/-0.33 (P=0.03). CONCLUSIONS: Our findings indicate that the HCV in situ RT-PCR assay may be helpful in the differentiation of recurrent hepatitis C disease from rejection. This may further help in the adjustment of immunosuppression.

Adult↗

Susceptibility of the liver and biliary tract to anaerobic infection in extrahepatic biliary tract obstruction. II. Liver function and morphology of the liver and biliary tract--an experimental study in rabbits.

Biliary tract infection with anaerobic bacteria (B. fragilis or Fusobacterium mortiferum) was produced in rabbits by common duct ligation (c.d.l.) 3 days prior to intravenous bacterial inoculation. Animals were investigated 1, 4 or 7 days after inoculation. Histopathological investigations included the liver, the common duct, and the gallbladder, while liver function was evaluated by bilirubin, alkaline phosphatase (AP), and L-alanine aminotransferase (GPT) in serum. Rabbits with c.d.l. and biliary tract infection were compared to rabbits with c.d.l. in which bacterial inoculation failed to produce infection, to inoculated rabbits without c.d.l., and to uninoculated rabbits with c.d.l. Anaerobic biliary tract infection in rabbits with c.d.l. caused a significant increase in liver abscesses, a significant increased infiltration with granulocytes in the gallbladder, and a significant increase in serum levels of bilirubin, AP, and GPT, but failed to produce signs of cholangitis in the liver and intramural abscesses in the gallbladder. A material is presented of normal values for bilirubin, AP, and GPT in serum in rabbits.

Alkaline Phosphatase↗

[Stimulation of DNA and RNA synthesis in the liver of hepatectomized rats by fractions of liver cytosol and conditioned media obtained during cultivation of rat liver explants].

Heat resistant fractions stimulating proliferation (PSF) were identified in the fractions of rat liver cytosol after 70% hepatectomy as well as in the medium harvested after cultivation of rat liver explants. The PSF augmented considerably the rate of DNA synthesis, it increased synthesis and transport of rapidly labelled RNA as well as the level of mitotic activity in rat liver tissue resected by 30% and 80%. Role of the PSF in regulation and acceleration of regeneration processes in the residual parts of liver tissue, remained after hepatectomy, is discussed.

Animals↗

Antibodies to liver cell membrane antigens in chronic active hepatitis (CAH). III. Partial characterization of the liver cell membrane antigens and comparison of reactivities in sera from patients with various liver diseases.

The reactivity of sera in liver diseases was investigated by an immunoradiometric assay for antibody to liver membrane antigens (LMAg). Serum reactivity was similar using glutaraldehyde treated hepatocytes from man and several animals, but the reactivity with human hepatocellular carcinoma cell lines was weak and irregular. F(ab)2 fragments from reactive sera were used in competitive inhibition experiments to ascertain whether anti-LMAg from different liver diseases reacted with the same or different antigenic determinants of the liver membrane. Sera from different patients with autoimmune CAH reacted predominantly with the same membrane components, as did sera from hepatitis B virus associated CAH (CAH-B); however sera from acute viral hepatitis A did not and, since such sera were reactive to LMAg in the assay, other membrane antigens are presumably involved. Enzymatic treatment of viable hepatocytes was performed to determine the nature of the reactive antigen(s): proteases or periodate resulted in reduced binding of reactive sera, but neuraminidase did not, suggesting that the LMAg recognized by CAH sera comprises species non-specific membrane glycoproteins. We conclude, on the basis of quantitative data from a immunoradiometric assay, that antibody to LMAg is more probably reactive than pathogenic, and that assessment of the antigenicity of specific membrane components is necessary.

Actins↗