bromochloroacetic-acid and Liver-Cirrhosis--Alcoholic

Mallory bodies (MBs) are characteristic morphologic features of alcoholic hepatitis but are also associated with non-alcoholic liver diseases including long lasting cholestasis, metabolic and neoplastic disorders. MBs contain in addition to keratins non-keratin components, including microtubule-associated (tau protein) and other not yet characterized proteins in an aggregated form. Aggregation of these components in the cell is promoted by posttranslational modifications, such as partial proteolysis, phosphorylation and cross-linking, and may result in functional and structural disturbances of the cell depending on the physiologic function of the components involved. Several enzymes responsible for these modifications are Ca(++)-dependent. Thus, disturbance of Ca(++)-homeostasis may play an essential role in the pathogenesis of MBs. In some structural aspects MBs closely resemble inclusions associated with degenerative disorders of the central nervous system, including Alzheimer's and Parkinson's disease. Studies on the pathogenesis of MBs, therefore, not only shed light on a peculiar type of liver cell injury but may also assist in the understanding of other chronic degenerative diseases, particularly those of the central nervous system.

Topics: Alzheimer Disease; Animals; Brain Diseases; Calcium; Homeostasis; Humans; Keratins; Liver; Liver Cirrhosis, Alcoholic; Liver Diseases; Liver Diseases, Alcoholic; Parkinson Disease; Protein Processing, Post-Translational; Transcription, Genetic

Mallory bodies are a morphological key feature of severe alcoholic liver cell injury (alcoholic hepatitis) and the morphological expression of dysregulation and derangement of the intermediate filament cytoskeleton of the hepatocyte. Their pathogenesis is still unclear. Studies on Mallory body formation may not only help to elucidate the mechanisms of liver cell injury associated with alcoholic hepatitis, but may also contribute to our understanding of the regulation and function of the intermediate filament cytoskeleton.

Topics: Animals; Chemical and Drug Induced Liver Injury; Colchicine; Cytoskeleton; Electrophoresis, Polyacrylamide Gel; Endoplasmic Reticulum; Fluorescent Antibody Technique; Griseofulvin; Hepatitis, Alcoholic; Humans; Isoantibodies; Isoelectric Focusing; Keratins; Liver; Liver Cirrhosis, Alcoholic; Mice; Microscopy, Electron; Rats

We describe the histopathologic features of 2 cases of biliary neoplasia with extensive intraductal spread arising in liver cirrhosis. The prevalence of this type of biliary neoplasia may be 0.4% from the review of 468 cases of cirrhotic liver. Histologic analysis revealed that the micropapillary proliferation of the atypical biliary epithelium composed of columnar cells with enlarged nuclei diffusely extended superficially from the septal intrahepatic bile duct to the reactive ductules associated with liver cirrhosis. Both cases exhibited prominent fibrous or sclerotic stroma near the biliary lesion. Immunohistochemical analysis revealed a characteristic cytokeratin and mucin expression pattern (CK7++, CK19++, CK20+, MUC1+/-, MUC2-, MUC5AC+, MUC6-). The tumor cytoplasm was focally positive for laminin gamma2 together with linear staining of the basement membrane. Proliferative activity confirmed by Ki67 staining was relatively high. Both patients were disease-free for 3 years after the operation. We believe that the possibility of biliary neoplasia with extensive intraductal spread should be considered to be a variant of biliary intraepithelial neoplasia.

Topics: Bile Ducts, Intrahepatic; Biliary Tract Neoplasms; Biomarkers, Tumor; Carcinoma, Hepatocellular; Cell Count; Cell Proliferation; Cholangiocarcinoma; Hepatitis C, Chronic; Humans; Immunoenzyme Techniques; Keratins; Liver Cirrhosis, Alcoholic; Liver Transplantation; Male; Middle Aged; Mucins; Neoplasms, Multiple Primary; Treatment Outcome

Fibrosis and nodular regeneration are the hallmarks of liver cirrhosis. To assess the degree of fibrosis and the severity of the structural changes affecting parenchymal and extraparenchymal components in liver cirrhosis, a computerized morphometric model has been applied to liver specimens from patients undergoing liver transplantation for primary biliary cirrhosis, posthepatitic and alcoholic cirrhosis. Fifty-eight hepatectomy specimens from patients undergoing liver transplantation for cirrhosis were analyzed: 17 alcoholic, 28 posthepatitic (HBV-related and HCV-related cirrhosis), and 13 primary biliary cirrhoses. Liver specimens were fixed in 10% neutral-buffered formalin and embedded in paraffin. Sections were stained with chromotrope-aniline blue method and monoclonal antibodies against cytokeratin 7 and CD31. Volume fractions of parenchymal compartment and fibrosis were stereologically determined on the specimens stained with chromotrope-aniline blue method. Volume fractions of portal bile ducts, proliferated bile ductules, and hepatocytes with biliary metaplasia were measured on cytokeratin 7 stains, while volume fractions of capillary units have been evaluated on CD31 staining. Volume fraction of fibrosis was higher in primary biliary cirrhosis than in the other disease-induced cirrhosis. The main differences were related to immunohistochemical staining. Volume fraction of hepatocytes with biliary metaplasia was higher in HCV-related cirrhosis, whereas volume fractions of biliary structures were more prominent in HBV-related cirrhosis. Primary biliary cirrhosis was characterized by a reduced number of bile ducts and by a wider expression of cytokeratin 7 into periportal hepatocytes. Capillary units were more prominent in primary biliary cirrhosis than alcoholic and posthepatitic cirrhosis. Our computerized morphometric model well describes and quantifies the morphological alterations of the liver and it could represent an adjunctive tool to evaluate the degree of dysplastic phenomena involving parenchymal and extraparenchymal compartments.

Topics: Adult; Female; Hepatectomy; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Keratin-7; Keratins; Liver; Liver Cirrhosis; Liver Cirrhosis, Alcoholic; Liver Cirrhosis, Biliary; Liver Regeneration; Liver Transplantation; Male; Microscopy; Middle Aged; Platelet Endothelial Cell Adhesion Molecule-1; Staining and Labeling

Not all alcoholic patients develop severe liver disease with fibrosis progressing to cirrhosis. It is of practical importance to determine whether some markers can predict progression of liver fibrosis.. We used a baboon model that mimics human alcoholic liver disease. Cytokeratin 7 and 19 expression and fat deposition were investigated in serial liver biopsies of 18 animals undergoing prolonged alcohol administration (range 2-17 years) and in four controls. Fibrosis was graded histologically and was also assessed quantitatively by image analysis.. Ten animals did not show a progression of liver disease even after 17 years of alcohol administration, but eight animals fed alcohol exhibited a progression of liver disease from no fibrosis or perivenular fibrosis to septal fibrosis or cirrhosis within 7 years. In normal liver, cytokeratin 7 and cytokeratin 19 immunostaining is restricted to bile duct cells. Hepatocellular cytokeratin 7 was observed only in those animals which progressed to more severe stages of fibrosis and it anticipated this progression by 4.2 years on average.. In alcohol-fed baboons, cytokeratin 7 staining of hepatocytes (but not cytokeratin 19, nor fat deposition) predicts with a high degree of sensitivity and specificity progression to more severe liver disease.

Topics: Animals; Bile Ducts; Disease Progression; Hepatocytes; Keratin-7; Keratins; Liver; Liver Cirrhosis, Alcoholic; Papio; Prognosis; Sensitivity and Specificity; Severity of Illness Index; Staining and Labeling; Time Factors; Tissue Distribution

In the present study we investigated tissue changes in palatine (PG) and labial (LG) minor salivary glands from individuals who had died of alcoholic hepatic cirrhosis, to characterize histopathological parameters of alcoholic sialosis, that may be used for differential diagnosis. Samples obtained from autopsies were processed using cytochemical techniques for mucosubstances and immunocytochemical labelling for cytokeratines, PS 100 and T-cells. Both PG and LG showed dilated excretory ducts with atrophic epithelium, which contained PAS+ metachromatic material and detached cells. Intra and interlobular ductal hyperplasia was present in some areas, mainly in PG. CK expression was heterogeneous in ductal cells, and negative in acinar cells. Most of the acinar cell nuclei were normal, but some of them were atypical in shape and distribution. Myoepithelial cells, serous demilunes and the basal region of the cells of the striated ducts expressed PS 100. In PG, 85% of the mononuclear infiltrates expressed T-cell markers, whereas in LG only 40% of these cells were T-cells. These findings, in addition to other histopathological parameters seen in previous studies, may be used as indicators for differential diagnosis with other gland pathologies.

Topics: Aged; Alcoholism; Case-Control Studies; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Keratins; Liver Cirrhosis, Alcoholic; Male; Middle Aged; S100 Proteins; Salivary Ducts; Salivary Gland Diseases; Salivary Glands, Minor; T-Lymphocytes

In the present study, the frequency and the distribution pattern of immunoreactive hepatocytic cytokeratin (CK) inclusions was investigated using the monoclonal antibody (MAb) CAM 5.2 detecting CKs 8, 18 and 19. The CK antigenicity of the inclusions was confirmed on frozen sections with MAbs for the CKs 7, 8, 17, 18 and 19. The frequency of hepatocytic CK aggregates was compared to the presence of non-alcoholic and alcoholic liver disease (ALD) as well as to the average all-year daily ethanol intake of 195 consecutive males subjected to medico-legal autopsy. Hepatocytic CK inclusions were infrequent in patients with normal liver histology, portal fibrosis and/or portal inflammation. In ALD, however, inclusions were observed in 35.6% of patients with fatty liver, in 63.2% of patients with alcoholic hepatitis, in 30.8% of patients with bridging fibrosis and in 60.0% of patients with liver cirrhosis. In all specimens studied, the inclusions were reactive only for CKs 8 and 18, CKs 7, 17, and 19 being unreactive. The frequency of inclusion bodies increased, paralleling increasing average all-year daily alcohol consumption. Compared to non-drinkers, a daily intake of between 40 and 80 g of absolute alcohol was associated with a statistically significantly (relative risk, RR = 6.6) increased risk of formation of aggregates. Similarly, daily consumption exceeding 80 g was related to increased (RR = 6.0) frequency of CK aggregates. The frequency of full-blown "classical" Mallory bodies (MBs) was, however, increased (RR = 8.9) only in patients with a daily intake exceeding 160 g.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Aged; Alcohol Drinking; Fatty Liver, Alcoholic; Hepatitis, Alcoholic; Humans; Immunohistochemistry; Inclusion Bodies; Keratins; Liver; Liver Cirrhosis, Alcoholic; Liver Diseases, Alcoholic; Male; Middle Aged

Severe ethanol-induced liver damage is characterized by fibrous dissociation of liver cell plates leading to many apparently isolated hepatocytes. Three-dimensional reconstruction, however, revealed hepatocytes that were surrounded by connective tissue as endpoints of "parenchymal pillars" or in association with liver cell plates and bile ductules. Double immunofluorescence studies displayed the expression of cytokeratin (CK) 7 in bile ducts, including bile ductules, but also in some hepatocytes still organized in liver cell plates. The other bile duct, typical CK, namely CK 19, was only detectable in few hepatocytes. However, the expression of CK 7 and/or CK 19 was less frequent in hepatocytes that were closely associated with bile ductules. CK 7 and CK 19 were also found in some, but not all, Mallory bodies. These observations indicate that the expression of these two CKs is neither related to a transformation of hepatocytes to bile duct-like structures ("ductal metaplasia") nor to the formation of Mallory bodies. Furthermore, double immunofluorescence studies revealed small groups of hepatocytes and bile ductules that were encircled by basement membrane material, thus suggesting the formation of "secretory units."

Topics: Aged; Bile Ducts; Cell Communication; Cell Membrane; Endoplasmic Reticulum; Extracellular Matrix; Female; Fibrosis; Fluorescent Antibody Technique; Humans; Image Processing, Computer-Assisted; Immunohistochemistry; Keratins; Liver; Liver Cirrhosis, Alcoholic; Liver Diseases; Male; Mesoderm; Middle Aged

Topics: Actin Cytoskeleton; Alcoholism; Animals; Cytoskeleton; Ethanol; Humans; Intermediate Filaments; Keratins; Liver; Liver Cirrhosis, Alcoholic; Microscopy, Electron; Rats

Antibodies raised against prekeratin intensely and specifically stain, in immunofluorescence microscopy, Mallory bodies ("alcoholic hyalin") present in livers of human alcoholics and griseofulyin-treated mice. The high sensitivity of this method allows the identification of small distinct cytoplasmic structures that are observed during early stages of Mallory body formation, especially frequent in the perinuclear cytoplasm, as well as during stages of Mallory body disintegration and disappearance, such as after withdrawal of the drug. In the latter situation, the prekeratin-containing small particles exhibit a characteristic pattern of arrangement in the hepatocyte periphery. Electron microscopy illustrates that such small bodies are heap-like aggregates of typical Mallory body filaments. Immunofluorescence studies with antibodies to isolated prekeratin polypeptides from bovine hoof or muzzle epidermis show that Mallory body filaments, in particular those in human liver, are immunologically more closely related to prekeratin of tonofilaments from living epidermal cells (stratum spinosum). The data indicate that Mallory bodies contain a pathologic form of prekeratin-like material. They also suggest that disorders of cytoskeletal structures of the intermediate-sized filament class are associated with specific diseases and can be visualized and characterized by immunofluorescence microscopy by using antibodies to constitutive proteins of such filaments.

Topics: Animals; Cytoplasmic Granules; Fluorescent Antibody Technique; Griseofulvin; Hepatitis, Alcoholic; Humans; Keratins; Liver Cirrhosis, Alcoholic; Liver Cirrhosis, Experimental; Mice; Protein Precursors