Isolation and Characterization of a Cellular Protein-Lipid Complex from Ascites Fluid Caused by Various Neoplasms

High concentrations of lipids in ascites fluid caused by peritoneal carcinomatosis have been described recently. Since their nature has not yet been clarified, we isolated ascitic lipids from 25 patients with various neoplasms for further characterization. After chromatography on Sephadex G-100 gels, the ascitic lipids were fractionated on a Biogel A-5m column in three peaks. The second and third peaks were identified as low and high density lipoproteins, which were most likely of plasmatic origin, and represented the major amounts of ascitic lipids. The first peak was eluted in the void volume, indicating a molecular weight over 5 million. It consisted, on the average, of 65.3% protein, 16.2% triglycerides, 7.4% phospholipids, and 7.0% cholesterol. In a CsCl gradient, this protein-lipid complex floated in the density range from 1.128 to 1.181 g/ml. Sodium dodecyl sulfate: polyacrylamide gel electrophoresis separated up to 11 protein subunits (Mr 29,000 to 97,000), and electron microscopy revealed globular particles of 36 to 64 nm in diameter. The macromolecular complex showed no immunological reaction against anti-{alpha}- and anti-ß-lipoproteins, but a single precipitation line against anti-liver-specific lipoprotein was seen. The biochemical characteristics of this protein-lipid complex proved to have a close relationship to liver-specific lipoprotein. It is most likely derived from cell membranes of the peritoneum detached by carcinomatosis

Point mutations of the P53 gene, human hepatocellular carcinoma and aflatoxins

The tumor suppressor p53 exerts important protective functions towards DNA-damaging agents. Its inactivation by allelic deletions or point mutations within the P53 gene as well as complex formation of wildtype p53 with cellular or viral proteins is a common and crucial event in carcinogenesis. Mutations increase the half-life of the p53 protein allowing the immunohistochemical detection and anti-p53 antibody formation. Distinct G to T point mutations in codon 249 leading to a substitution of the basic amino acid arginine by the neutral amino acid serin are responsible for the altered functionality of the mutant gene product and were originally identified in 8 of 16 Chinese and 5 of 10 African HCC patients. Both groups are frequently exposed to mycotoxin contaminations of their food. Today an average P53 gene mutation rate of 25% is assumed for high-aflatoxin B1-exposure regions. This is double the rate observed in low-aflatoxin B1-exposure countries. Although many HCC patients displaying P53 mutations also suffer from HBV infection, which itself can lead to rearrangements of P53 coding regions or induce the synthesis of viral proteins possibly interacting with p53, the specific G to T transversion within codon 249 of the P53 gene seems to directly reflect the extent of aflatoxin B1 exposure

Liver resection or combined chemoembolization and radiofrequency ablation improve survival in patients with hepatocellular carcinoma

Background/ Aims: To evaluate the long-term outcome of surgical and non-surgical local treatments of patients with hepatocellular carcinoma (HCC). Methods: We stratified a cohort of 278 HCC patients using six independent predictors of survival according to the Vienna survival model for HCC (VISUM- HCC). Results: Prior to therapy, 224 HCC patients presented with VISUM stage 1 (median survival 18 months) while 29 patients were classified as VISUM stage 2 (median survival 4 months) and 25 patients as VISUM stage 3 (median survival 3 months). A highly significant (p < 0.001) improved survival time was observed in VISUM stage 1 patients treated with liver resection ( n = 52; median survival 37 months) or chemoembolization (TACE) and subsequent radiofrequency ablation ( RFA) ( n = 44; median survival 45 months) as compared to patients receiving chemoembolization alone (n = 107; median survival 13 months) or patients treated by tamoxifen only (n = 21; median survival 6 months). Chemoembolization alone significantly (p <= 0.004) improved survival time in VISUM stage 1 - 2 patients but not (p = 0.341) in VISUM stage 3 patients in comparison to those treated by tamoxifen. Conclusion: Both liver resection or combined chemoembolization and RFA improve markedly the survival of patients with HCC

Design, delivery and efficacy testing of therapeutic nucleic acids used to inhibit hepatitis C virus gene expressionin vitroandin vivo

interior, view of lobby and interior arches, ca. 198

Insertional activation of mevalonate kinase by hepatitis B virus DNA in a human hepatoma cell line

Insertional mutagenesis of growth related genes by hepatitis B virus (HBV) DNA is presumed to play a role in hepatocarcinogenesis. Here, we report on insertional activation of the mevalonate kinase (MK) gene in the human hepatoma cell line PLC/PRF/5. Integration of HBV DNA dissociated the promoter and upstream regulatory elements of the gene from its coding sequences. This led to the over-expression of hybrid transcripts arising from an HBV promoter and the consequent over-production of functionally active mevalonate kinase. MK phosphorylates mevalonate, a major intermediate in the branched cholesterol/isoprenoid biosynthetic pathway. Isoprenylation is crucial to the functions of cellular proteins related to growth control, including the proto-oncogene ras. As the enzymes of these biosynthetic pathways are regulated at multiple points by negative feedback, both transcriptionally and at the protein level, the results discussed here support the idea that aberrant growth could result from deregulated overexpression of MK and, perhaps, other enzymes in the cholesterol pathway. These results invoke novel mechanisms by which cell transformation might occur.</p