Endoscopic management of pancreaticopleural fistulas: a report of three patients.

Pancreaticopleural fistulas are a rare complication of acute or chronic pancreatitis, and are usually treated by surgery. We report three patients whose pancreaticopleural fistulas were successfully treated by endoscopic retrograde cholangiopancreatography and drainage (stenting, nasopancreatic drainage). In one patient a pancreatic pseudocyst persisted despite successful initial closure of the leak using this method and, as it was also suspected to be infected, additional drainage of the pseudocyst was required. Endotherapy of pancreaticopleural fistulas could obviate the need for surgery when conventional medical treatment has failed in this condition.

Interesting RI accumulation in hepatic images with Tc-99m GSA SPECT scintigraphy in idiopathic portal hypertension.

The authors report a case of idiopathic portal hypertension (IPH), which showed interesting RI accumulation on Single Photon Emission CT (SPECT) images, with Tc-99m galactosyl human serum albumin (GSA) scintigraphy. Accumulation of Tc-99m GSA was decreased in the periphery of the liver where strong enhancement was revealed only in the arterial phase on dynamic CT. These findings imply that portal flow is decreased in the periphery of the liver where arterial flow is dominant. It was thought that secondary reduced activity of GSA due to a decrease in portal flow results in reduced radioactivity in the periphery of the liver in IPH. This interesting accumulation of Tc-99m GSA may be one of the sign of IPH.

Cynomolgus monkey liver aldehyde oxidase: extremely high oxidase activity and an attempt at purification.

Aldehyde oxidase (EC 1.2.3.1) in monkey (Macaca fascicularis) liver was characterized. Liver cytosol exhibited extremely high benzaldehyde and phthalazine oxidase activities based on aldehyde oxidase, compared with those of rabbits, rats, mice and guinea pigs. Monkey liver aldehyde oxidase showed broad substrate specificity distinct from that of the enzyme from other mammals. Purified aldehyde oxidase from monkey liver cytosol showed two major bands and two minor bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). These bands were also observed in Western blotting analysis using anti-rat aldehyde oxidase. The molecular mass of the enzyme was estimated to be 130-151 kDa by SDS-PAGE, and to be about 285 kDa by HPLC gel filtration. The results suggest that isoforms of aldehyde oxidase exist in monkey livers.

Purification and some properties of hamster liver aldehyde oxidase.

Aldehyde oxidase was purified from hamster liver cytosol by ammonium sulfate fractionation, chromatography on DEAE-cellulose and Phenyl-Toyopearl, and HPLC-gel filtration on TSK-gel G3000SW(XL) column. The purified enzyme was homogeneous by the criterion of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Its molecular weight was determined to be 144800 by SDS-PAGE and 288000 by HPLC gel filtration. The isoelectric point was pH 5.1. The apparent Km and Vmax for benzaldehyde and 2-hydroxypyrimidine were 19.0 and 4.4 microM, and 165 and 211 nmol/min/mg protein, respectively. The benzaldehyde oxidase activity was markedly inhibited by menadione and chlorpromazine. The substrate specificity was different from those of the enzymes from other animals.

Rupture of a pancreatic pseudocyst into the portal venous system.

Pseudocyst formation is a well-known complication of acute or chronic pancreatitis. We report a case in which pseudocyst ruptured into the splenic and portal veins.

An automated new technique for scoring the rodent micronucleus assay: computerized image analysis of acridine orange supravitally stained peripheral blood cells.

We developed an automated image analysis system to obtain objective data for the rodent peripheral blood micronucleus assay with acridine orange (AO) supravital staining. The system was able to identify micronucleated reticulocytes (MNRETs) and to evaluate inhibition of bone marrow cell proliferation by measuring the reticular area of reticulocytes (RETs). We also developed automated equipment to produce homogeneous acridine orange-coated glass slides. This study was designed to compare automated scoring with manual scoring using 4 model clastogens and 2 mouse strains. The MNRET incidence induced by each clastogen was similar for automated and manual scoring, and there was good correlation (r=0.92) between the methods. In addition, an index of bone marrow toxicity based on the reticular area of RETs was compared to the conventional index (% of polychromatic erythrocytes (PCE) to total erythrocytes; PCE ratio) and was similar. The results indicated that our technique for computer-assisted image analysis for the micronucleus assay with AO supravitally stained peripheral blood RETs was comparable to conventional microscopic scoring, and it was superior in objectivity and statistical power.

Altered cytokeratin expression and differentiation induction during neoplastic transformation of cultured rat liver cells by nickel subsulfide.

Rat liver T51B cells were maintained in the presence of low concentrations of Ni(II) derived from alpha Ni3S2 for 3-15 months in culture in order to monitor cytokeratin, differentiation, and transformation patterns. Nickel exposures caused irreversible, heritable juxtanuclear aggregates of cytokeratin CK55, which increased in size and complexity with prolonged nickel exposure, eventually resembling Mallory bodies and expressing glutamyltransferase. Altered cytokeratin expression was accompanied by induction of differentiation, with markers of both bile ductular cells and hepatocytes, such as induction of cytokeratin polypeptides CK39 and CK49, cell morphology, and cytokeratin filament network changes; whereas control cultures similarly maintained for long periods in culture remained unchanged. Altered cytokeratin expression was also accompanied by acquisition of transformation markers--loss of density dependence, progression toward calcium independence, and (benign) growth in nude mice. Observed cytokeratin aberrations may be a factor in nickel carcinogenesis, in view of the known affinity of the metal for cellular structural proteins, especially keratin, which play a role in maintenance of cell behavior.

Interaction of intermediate filaments with nuclear lamina and cell periphery.

Ultrastructural observations of the cytoskeleton suggest that the connection of the intermediate filaments (IFs) to actin microfilaments (MFs) at the plasma membrane and the nuclear lamina inside the nuclear membrane link signals received at the cell periphery to the nucleus. When these connections are viewed in three dimensions using detergent extracted cytoskeletal preparations from tissue cultures or slices made from tissue, the IFs are seen to run without interruption from the cell periphery to the nucleus and back. The IFs form side to side connections with the nuclear lamina and pore complexes. The nucleus and the centrioles are supported and held suspended in these extracted cells where all organelles and cytosol have been removed. The IFs are particularly dense in the ectoplasm where they form a sheet and provide the scaffolding which maintains the shape of the extracted cells. The IFs in the ectoplasm are attached to desmoplakin at cell-cell desmosome adhesions and to MFs where the cells are attached to the fibronectin substratum possibly through integrin linkages at adhesion plaques. This was graphically shown by immunogold labelling of IF cells treated with nickel. In this way, it was possible to visualize the loss of the cell-cell connections at desmosomes and the disruption of the IF-MF connections in the ectoplasm. The MFs after losing their connections with the IFs, redistribute to cover the entire cell periphery. The nickel treatment of primary liver cell cultures lead to the loss of several functions including formation of the bile canaliculus, the ability to secrete fluorescein diacetate and the ability to take up horseradish peroxidase (HRP) by endocytosis. These observations support the conclusion that the IF-MF connections at the cell periphery provide both structural and functional polarity of the liver cells including uptake and secretion and the formation of bile canaliculi.

Cytokeratin intermediate filaments of rat hepatocytes: different cytoskeletal domains and their three-dimensional structure.

A new method of visualizing the three-dimensional architecture of the cytokeratin filaments of the intact rat hepatocyte in situ has been achieved. Frozen sections of liver cut 10 micron thick were serially extracted to remove all elements of the cells except the intermediate filaments. Parallel sections were stained with monoclonal antibodies to the two main cytokeratins found in bile duct and liver cells. Immunofluorescent antibody and immunogold electron microscopy techniques were used to identify the proteins morphologically. Several new observations resulted from these studies. The pericanalicular sheath of intermediate filaments was visualized using steropairs as an uninterrupted branching tubular structure composed of cytokeratins located in the cell cortex of adjacent hepatocytes. Intermediate filaments in the cell cortex formed a distinct sheet of matted filaments which enveloped the entire hepatocyte. The cortical intermediate filaments were in continuity with the pericanalicular sheath and the filaments located within the cytoplasm. The intermediate filaments are attached to the centrioles and appeared to tent the nuclear lamina-pore complex at points of contact. Monoclonal antibodies to rat liver intermediate filament cytokeratins (CK49 and CK55) each stained intermediate filaments located in the cell cortex, within the cytoplasm and at the nucleus. By immunogold staining, some of the intermediate filament filaments were shown to contain both cytokeratins. Filaments which did not stain were thought to be either actin at the cell periphery or nuclear lamins around the nucleus. It is concluded that the cytokeratins form a specialized framework for the cell cortex, canaliculus, centrioles and the nucleus of hepatocytes. The filaments run continuously throughout the cytoplasm without terminating.

Selective rearrangement of cytokeratin filaments in cultured liver epithelial cells induced by nickel.

Rearrangements of intermediate filaments (IFs) in the liver cells involve cytokeratins, i.e., Mallory body formation. However, rearrangements of IFs observed in cell culture characteristically involve vimentin or both vimentin and cytokeratin. We report here that nickel treatment of a liver epithelial cell line (T51B) in vitro selectively induced cytokeratin filaments to accumulate in a juxtanuclear location as the cells rounded up. After withdrawal of nickel from the culture medium, vimentin filaments remained attached to the cell periphery as the cells spread out again, but the cytokeratin filaments remained aggregated in a perinuclear position without reestablishing all peripheral connections. Thus, the rearrangement of cytokeratin filaments involved partial loss of the peripheral attachments in this model.

Changes in the cytokeratin intermediate filament cytoskeleton associated with Mallory body formation in mouse and human liver.

Mouse and human extracted liver tissue were examined by indirect immunofluorescent staining and transmission electron microscopy in order to study the alteration of cytokeratin intermediate filaments associated with Mallory body formation. Frozen sections of griseofulvin-fed mouse liver and human liver of primary biliary cirrhosis and primary sclerosing cholangitis were extracted by Triton X-100 and nuclease. Indirect immunofluorescent staining was performed by using anticow hoof keratin antibody for mouse liver and anti-human epidermal keratin antibody (AE1 and AE3) for human liver. Transmission electron microscopy was also performed on extracted and critical point-dried samples. The griseofulvin-fed mouse hepatoma cells showed four different types of altered staining pattern based on the indirect immunofluorescent staining of the cytoplasm and Mallory bodies: Type I--cytoplasm(+), Mallory body(-); Type II--cytoplasm(+), Mallory body(+); Type III--cytoplasm(-), Mallory body(+), and Type IV--cytoplasm(-), Mallory body(-). Types I and III were predominant, however, some hepatoma cells which contain Mallory bodies revealed bright cytoplasmic staining (Type II). The nuclear rims were strongly stained. In human liver, AE1 stained Mallory bodies and the bile duct epithelium intensely, but did not stain normal hepatocytes. AE3 mainly stained Mallory bodies and normal hepatocytes, but also stained bile duct epithelium weakly. Indirect immunofluorescent staining for human liver showed the same staining patterns as found in mouse liver, except that Type IV was not observed. Although many hepatocytes which contained Mallory bodies did not react with either of these two antibodies (Type III), some of the hepatocytes were stained, not only with AE3, but also with AE1 (Type II).(ABSTRACT TRUNCATED AT 250 WORDS)

Connections of intermediate filaments with the nuclear lamina and the cell periphery.

We investigated the relationship between intermediate filaments (IFs) and other detergent- and nuclease-resistant filamentous structures of cultured liver epithelial cells (T51B cell line) using whole mount unembedded preparations which were sequentially extracted with Triton X-100 and nucleases. Immunogold labelling and stereoscopic observation facilitated the examination of each filamentous structure and their three-dimensional relationships to each other. After solubilizing phospholipid, nucleic acid and soluble cellular protein, the resulting cytoskeleton preparation consisted of a network of cytokeratin and vimentin IFs linked by 3 nm filaments. The IFs were anchored to and determined the position of the nuclear lamina filaments (NLF) network and the centrioles. The NLF was composed of the nuclear lamina filaments measuring 3-6 nm in diameter which radiated from and anchored to the skeleton of the nuclear pores. The IFs located in the nuclear region appeared to be interwoven with the NLF. At the cell surface, the IFs seemed to be attached to the putative actin filament network. They formed a focally interrupted plexus-like structure at the cell periphery. Fragments of vimentin filaments were found among the filamentous network located at the cell surface, and some filaments terminated blindly there.

Pathology of acute hepatitis A in humans. Comparison with acute hepatitis B.

Little is known about the pathologic characteristics of viral hepatitis A in humans. The authors compared the histologic features in liver biopsy specimens taken within 30 days of the onset of illness from 15 patients with hepatitis A and 14 patients with acute hepatitis B. In both hepatitis A and B, liver cell damage and necrosis were diffusely located, with accentuation in the centrilobular and midzonal areas in which ballooning degeneration and variation in cytoplasmic staining quality were observed frequently. One case of epidemic hepatitis A showed prominent periportal liver cell necrosis with inconspicuous centrilobular liver cell alterations. Kupffer cell mobilization was mild in hepatitis A, but more striking in hepatitis B. The portal inflammation was more pronounced and rich in plasma cells in hepatitis A than in hepatitis B. In summary, there were no major differences in the pathologic features of acute hepatitis A and B as sampled within 30 days of the onset of illness.