Microtubules are required for cytokeratin aggresome (Mallory body) formation in hepatocytes: an in vitro study

Mallory bodies are cytokeratin-ubiquitin aggresomes that form in hepatocytes in many different chronic liver diseases. One of the key components in aggresome formation, not yet investigated in Mallory body formation, is the role of microtubules. An in vitro tissue culture assay is required to test for microtubule involvement in Mallory body formation so that Mallory body formation can be observed in the presence or absence of microtubule-disrupting agents. In this report, a new model of in vitro Mallory body formation was developed, which uses cultured hepatocytes isolated from drug-primed mice. When hepatocytes were incubated in the presence of antimicrotubule agents, they failed to form Mallory bodies. It is concluded that intact microtubules are required for Mallory body formation.     

M-30 and 4HNE are sequestered in different aggresomes in the same hepatocytes

M-30 and 4HNE adducts are two markers of active liver disease. M-30 is a serologic marker and 4HNE adducts are histologic markers. M-30 is a marker for apoptosis because it is a fragment of cytokeratin-18 left over from proteolysis by caspase 3. 4HNE is a marker of oxidative stress because it results from lipid peroxidation. Both markers are commonly found in nonalcoholic steatohepatitis and in alcoholic hepatitis. Liver biopsies from patients with steatohepatitis, 11 alcoholic and 11 non-alcoholics were stained for 4HNE and M-30. Almost all of the biopsies in both groups showed 4HNE- and M-30-positive aggresomes in hepatocytes. Mallory Denk bodies (MDB) stained variably positive for M-30, whereas 4HNE was present in aggresomes independent of MDBs. However, they were sometimes located in hepatocytes which also contained MDBs as shown by confocal microscopy of double stained biopsies. The results indicate that the formation of M-30 and 4HNE aggresomes occurs through different pathways of liver cell injury in both types of steatohepatitis.     

Peculiar changes in Rosenthal fibres in an atypical astrocytoma

A 50-year-old female patient died of an untreatable glioma apoplecticum. At autopsy a strongly vascularized glial tumour was found. The criteria for malignancy according to the WHO classification were only partially fulfilled by this tumour which displayed morphological features of an astrocytoma but could not be further subclassified. By light microscopy, angioma-like vascular proliferations, large cells with brightly eosinophilic cytoplasm, and small cells with hyperchromatic nuclei were found. Most large cells had vesicular, excentrically placed nuclei and contained fibrillary whorls or amorphous, irregular cytoplasmic inclusions. By immunohistochemical staining, using antibodies to glial fibrillary acidic protein (GFAP) the fibrillary whorls were identified as aggregates of glial filaments. The amorphous inclusions lacked GFAP immunoreactivity and appeared in the electron microscope as electron dense material surrounded by a dense network of glial filaments. The abnormal perikaryal inclusions of these atypical astrocytoma cells appeared to be peculiar alterations of Rosenthal fibres closely mimicing Mallory bodies.     

Clinicopathological characteristics of hepatocellular carcinoma bearing Mallory bodies: an autopsy study

Abstract— Mallory bodies are known to occur in hepatocellular carcinoma. The simple question whether or not there are any clinicopathological features characterizing Mallory body-positive hepatocellular carcinoma remains unresolved to date. The present study of 200 consecutive autopsy cases of hepatocellular carcinoma showed several important differences between 49 cases bearing Mallory bodies and 151 cases bearing no Mallory bodies in carcinoma cells. The patients in the former group were older, showed a higher association rate of liver cirrhosis, and their liver weight was lighter. As to the gross pathology of hepatocellular carcinoma, the nodular type was relatively frequent in Mallory body-positive hepatocellular carcinoma, while the massive and diffuse types were relatively frequent in Mallory body-negative cases. The frequency of extrahepatic metastases in the Mallory body-positive group was lower than that in the Mallory body-negative cases. The reasons for these differences remain speculative.     

Keratin inclusions alter cytosolic protein localization in hepatocytes

Aim:  Mallory bodies have been observed in various liver diseases, however, the precise mechanism and significance of these structures have yet to be determined.
Methods:  Previously we reported on the redistribution of cytosolic proteins to keratin inclusions in mutant keratin 18-transfected cells. In this study, we treated green fluorescent protein-tagged wild-type keratin 18-transfected cells with several proteasome inhibitors and performed immunofluorescent analyses.
Results:  Proteasome inhibitors induced intracellular keratin inclusions, and desmoplakin, zonula occludens-1 and β-catenin were relocated to keratin inclusions, while theintegral membrane proteins were intact. The cytosolic proteins, 14-3-3 ζ protein and glucose-6-phosphate dehydrogenase were also relocated to inclusions. Moreover, E-cadherin, a basolateral membrane protein, was present on both the apical and basolateral domains in inclusion-containing cells.
Conclusion:  These data are identical to those in the mutant keratin 18 transfection study and suggest that keratin inclusions induced by different treatments affect localization of various cytosolic components, which may influence cellular functions performed by these proteins.     

Diffuse Mallory Bodies in the Liver, Diffuse Lewy Bodies in the Brain and Diffuse Fat Replacement (Lipomatous Pseudohypertrophy) of the Pancreas in a Patient with Juvenile Parkinson's Disease

A 38-year-old male patient with the juvenile variant of Parkinson's disease, in whom onset had occurred at the age of 24 yr, was autopsied. There were no clear symptoms of pancreatic or hepatic insufficiency during the entire clinical course. The only notable features were a slightly delayed decrease of the blood glucose level in an oral glucose tolerance test, slightly elevated levels of serum alkaline phosphatase and serum lactate dehydrogenase, and episodic loose stools. Autopsy revealed uniform enlargement of the pancreas due to massive fat replacement (lipomatous pseudohypertrophy): the exocrine glandular elements showed marked atrophy and loss, while the islets of Langerhans were preserved. The liver exhibited a histology closely mimicking alcoholic hepatitis associated with the diffuse presence of Mallory bodies (MBs), possibly indicative of a disturbance of protein metabolism. The nervous system showed the diffuse presence of Lewy bodies (LBs) in the cerebrum in addition to the ordinary lesions of Parkinson's disease. Although the etiopathogenesis of none of these three lesions has been well elucidated, common epitopes of MBs and LBs have recently been demonskated. Therefore, the present case study suggests that a specific underlying toxic agent may cause diffuse LBs in the brain on the one hand, and diffuse MBs in the liver and lipomatous pseudohypertrophy of the pancreas on the other. Acta Pathol Jpn 42: 826–831, 1992.     

Infiltrating Polymorphonuclear Leukocytes and Apoptotic Bodies Derived From Hepatocytes but Not From Ballooning Hepatocytes Containing Mallory Bodies Show Nuclear DNA Fragmentation in Alcoholic Hepatitis

Background: The mechanism of liver cell death in alcoholic hepatitis is still controversial. There is evidence that polymorphonuclear leukocytes (PMNs) which infiltrate liver parenchyma are involved in liver cell death. Methods: We employed terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphatedigoxigenin nick end labeling (TUNEL) method to determine the cells that underwent apoptosis in liver specimens obtained from 1 normal liver, 23 livers with nonalcoholic liver disease, and 11 livers with alcoholic liver diseases (4 alcoholic hepatic fibrosis, 4 alcoholic hepatitis, and 3 alcoholic cirrhosis). The Fas, bcl-2, and CD15 antigens were immunolocalized to analyze the relationship between these antigens and apoptosis. Results: Few TUNEL-positive reactions were found in normal liver. Apoptotic bodies in acute and chronic viral hepatitis were found to be TUNEL-positive. TUNEL-positive materials were also abundant in the cytoplasm of macrophages at the area of parenchymal collapse in viral liver diseases. Mononuclear cells that infiltrated liver parenchyma and portal tracts occasionally showed TUNEL-positive reactions, which reflected the activation-induced apoptosis of T lymphocytes. In alcoholic liver diseases, the incidence of apoptotic bodies derived from hepatocytes varied from 0 to 1.00/mm². In addition, a considerable portion (0.2-16.5%) of infiltrating PMNs in alcoholic hepatitis were TUNEL-positive. In contrast, the nuclei of ballooning hepatocytes that contained Mallory bodies revealed few positive TUNEL reactions. Interestingly, the density of CD15-positive PMNs was well correlated with peripheral white blood cell counts (r = 0.842, p < 0.005). Conclusions: PMNs that infiltrate liver parenchyma show an activation-induced cell death-like phenomenon in alcoholic hepatitis, like activated T cells that infiltrate in viral hepatitis. This apoptosis phenomenon may follow the degranulation of PMNs, which may cause a necrosis of ballooning hepatocytes that contain Mallory bodies through satellitosis.     

Mallory Weiss syndrome is not associated with hiatal hernia: a matched case–control study

Background/objective: Hiatal hernia is considered to be a predisposing factor to develop Mallory-Weiss Syndrome (MWS). No large case–control studies verifying this hypothesis have been conducted.

Methods: We reviewed all esophagogastroduodenoscopies with findings of MWS (n?=?2342) in a national database and compared with age and gender-matched controls (n?=?9368). Demographics, endoscopic characteristics and presence of a hiatal hernia were compared between both groups. Average age was 56.7?±?18.6 years, and 72.4% were male.

Results: Hiatal hernia was more common in controls, and no significant difference was seen in a multivariate analysis.

Conclusion: Dynamic changes inducing mucosal tension are more relevant determinants to develop MWS than gastro-esophageal junction location alone.     

Alcoholic hepatitis with leukemoid reaction after surgery

Alcoholic hepatitis (AH) is a clinicopathologic syndrome resulting from an excessive intake of alcohol. Leukemoid reactions (LRs) are characterized by a strikingly elevated granulocyte count over 40,000-50,000 cells/mm³. Although a leukocytosis of 15,000-18,000 cells/mm³ is frequently seen in AH, LRs are rare in this context. AH-associated LRs are a sign of poor prognosis and have a high mortality. A 64-year-old male with a history of heavy alcohol intake underwent a right hemicolectomy for cecal carcinoma. Preoperative laboratory data were normal with the exception of an albumin of 2.1 g/dL. Liver biopsies that were taken because of a nodular appearance revealed micronodular cirrhosis, steatohepatitis, and Mallory bodies. Postoperatively, the patient developed a leukocytosis that progressively increased to 72.6 cells/mm³. He also developed signs of impaired hepatic and renal function. Extensive workup failed to reveal a source of infection. A trial of intravenous antibiotics had no impact on the leukocytosis. Methylprednisolone at a dose of 40 mg IV daily was started on postoperative day 9. The patient experienced a progressive decline in white blood count (WBC), which reached 25.2/mm³ on postoperative day 14. However, he died on postoperative day 16. We conclude that the patient had AH-associated LR in the postoperative period, but died despite successful treatment of the LR with steroids. Presented at the 2005 American Hepato-Pancreato-Biliary Association Congress, Hollywood, Florida, April 14–17, 2005 (poster presentation).