161 BILE ACIDS REDUCE THE BINDTRG AND UPTAKE OF ENDOTOXIN BY CULTURED RAT KUPFFER CELLS
H. Van Bossuyt,C. Desmaretz, G.B. Gaeta, E. Wisse. Lab. Cell Biology, Free University Brussels, Laarbeeklaan 103, 1090 Brussels-Jette,Belgium
According to l i terature ,25% to 69% of the p a t i e n t s with obstructivejaundiceshowendo- toxemia. There~sconsiderable evidence that endotoxemia is the underlying cause of some complications seen in obstructive jaundice. Endotoxemia and the outcome of jaundiced patients af ter surgery are re lated, since a l l patients who died af ter surgery for jaun- dice showed preoperative endotoxemia.Bile acids (BA) prevent the absorption of endotoxin ( l ipopolysaccharide,LPS) from the intest ines into the portal blood.The absence of BA in the intest ines in obstructive jaundice may contr ibute to portal endotoxemia.LPS would normally be cleared by the Kupffer ce l ls (Kc) . In th is study the ef fect of BA on the up- take of LPS by Kc was investigated in v i t ro . Kc were isolated from male Wistar rats and put in cul ture.Cul tured Kc were incubated with t r i t i a t e d LPS in presence of d i f fe rent concentrations of BA (85% TCA, 15% TDCA) or not. The percentage of cel l -associated LPS increased with time an~ reached a plateau af ter about 2h of incubation.About 1.2/~g LPS was associated with I0 Kc at th is time i n t e r v a l . l n presence of 0.3, 0.6 and I/~nol BA/ml the cel l -associated LPS was 5%, ]3% and 29% lower than in control cultures.The binding at O°C was about 25% lower in presence of ~}~mol BA/ml.Preincubation of Kc with ~mol BA per m] did not the binding and uptake of LPS af ter removal of the BA.The rate of secre- t ion of rad ioac t iv i ty by Kc was not influenced by the presence of BA. From these observations we conclude that BA inf luence the binding and uptake of LPS by Kc. We therefore consider i t l i k e l y that , i i~patients with obstructive jaundice, serum BA may account for the sp i l l over of portal endotoxin into the systemic blood.
162 DES-GAMMA-CARBOXYPROTHROMBIN ACTIVITY IN HEPATOCELLULAR CARCINOMA
V. Vargas, J.Monasterio , J.Mir*, A.Gonz~lez, R.Esteban, J.Guardia .
Liver Unit. Department of Internal Medicine. Hospital "Vall d'Hebron". Universi dad Autdnoma and * Department of Hematology. H. "Vall d'Hebron". BARCELONA.
The presence of dlfferent forms of prothrombin in patients with hepatocellular carcinoma (HCC) has suggested that these proteins may be useful in its diagnosis. The aim of our study
was to establish the value of des-gamma-carboxyprothrombin (DCP) as a marker of HCC. We have measured the actlvity in plasma in patients with such illnes and in patients with other liver diseases. The DCP levels were compared with those of alphafetoproteih (AFP)
We studied 20 patients with HCC, 55 with hepatic cirrhosis, 10 with metastatic liver di- sease and 14 with other liver diseases. We also measured DCP in 30 asymptomatic blood donors.
DCP functional activity assay was performed using staphylocoagulase on citrated plasma af- ter bentonite and aluminum adsorption. The thrombin-coagulase formed was measured by the in-
cre&se in optical density of a chromogenic substrate. The results were expressed in mU. AFP was measured by RIA and the results were expressed in ng/ml.
The cutoff point of DCP level was stated at 4.5 mU. DCP was positive in 17 of 20 patients with HCC (85%), while it was negative in all the patients with hepatic cirrhosis and with other non-neoplastic liver diseases. Only one of i0 patients with metastasic liver diseases presented positive levels. The sensivity of the assay was 85% and the specificity was 98%. The AFP ( more than 40 ng/ml) was positive in 17 HCC (85%), with a specificity of 91%. Two of three patients with negative AFP had positive DCP.
The DCP activity assay using staphylocoagulase is a new method, that does not require radio isotopes and that has similar sensitivity as AFP with a higher specificity. This assay may be useful as a biological marker of HCC.