Bile acid contents in isolated rat hepatocytes were determined by gas-liquid chromatography-mass spectrometry-selected ion monitoring with the use of deuterium-labeled internal standards. This allowed us first to monitor the actual amounts of not only major but also minor bile acid components present with sufficient sensitivity and specificity and to follow the changes of individual bile acids in cultured rat hepatocytes simultaneously. In freshly isolated rat hepatocytes, cholic and beta-muricholic acids were the major components, comprising 35 and 46% of the total bile acids, respectively. These two bile acids were found to be most actively synthesized during the first 2 h of incubation and continued to increase thereafter for up to 6 h (the end of the period studied). In contrast, chenodeoxycholic and alpha-muricholic acids, which are the precursors of beta-muricholic acid, showed slight increases only in the first hour of incubation and decreased thereafter. These results suggested that the conversion to beta-muricholic acid from chenodeoxycholic acid via alpha-muricholic acid occurred rapidly in cultured rat hepatocytes. The secondary bile acids such as deoxycholic, hyodeoxycholic, and 3 alpha, 12 beta-dihydroxy-5 beta-cholanoic acids declined steadily from the start of incubation, which supported the findings that further hydroxylation of these dihydroxy bile acids occurs in rat liver.
Kupffer cells (KC) were isolated from the liver of guinea pigs, rats, and mice using enzymatic digestion with collagenase, followed by differential centrifugation and plastic adherence. Purity of the isolated KC was 96.0±2.2, 97.2±2.1, and 96.0±2.3 per cent in guinea pigs, rats, and mice respectively. These isolated KC were tested for migratory response to bacterial factor, which is one of the representative chemotactic factors for inflammatory macrophages, using a modified Boyden chamber technique. KC from the three animal species similarly migrated to the bacterial factor. The migratory response of the KC to the bacterial factor is due to chemotaxis but not chemokinesis. These results show the possibility that KC may recognize a chemoattractant and directionally migrate to it.
One hundred and seventy cases of cancer of digestive tract were separated into three groups, advanced gastric cancer, biliary and/or pancreatic cancer and colorectal cancer. The presence of liver metastasis in each group was studied preoperatively by either liver scintigraphy or celiac and superior mesenteric angiography. In advanced gastric cancer and colorectal cancer, false negative ranged between 6.7-12.7 per cent by both scintigraphy and angiography. In biliary tract and/or pancreatic cancer, the ratio of false negative was significantly higher i.e. 18.8-22.6 per cent, which suggests the difficulty in diagnosing liver metastasis correctly in this group. The most frequent occurrences of false positive either in scintigraphy or angiography were those of masses of less than 2 cm in diameter. The difference in correct diagnosis ratio for liver metastasis between scintigraphy and angiography in all three groups was only 2.9 per cent. Hence, liver scintigraphy seems to be preferable for the purpose of detecting liver metastasis prior to surgery, since further angiographical examination entails irradiation, possible complications and economic factors.
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