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The high incidence of biliary tract carcinoma in patients with anomalous pancreaticobiliary ductal junction (APBDJ) with or without choledochal cyst (CC) has been well documented. Twenty-two patients with APBDJ were divided into three groups: Group A, four patients not associated with CC and biliary tract carcinoma; Group B, 13 patients with CC but without biliary tract carcinoma; and Group C, five patients with biliary tract carcinoma (four with and one without CC). Profiles of bile acids in the gall-bladder and/or common bile duct were analysed in these patients and compared with those in the control patients with cholecystlithiasis to examine the hypothesis that the levels of deoxycholic acid (DCA) and lithocholic acid (LCA) are elevated in patients with APBDJ because these secondary bile acids are mutagenic. Bile acids were quantified by gas-liquid chromatography. Total bile acid concentration in the gall-bladder bile was significantly lower in any group with APBDJ than that of controls. In the gall-bladder, increased proportion of chenodeoxycholic acid (CDCA) in Group A and B, decreased proportion of DCA in Group B and increased proportion of cholic acid (CA) in Group C were found in bile. In the bile duct, total bile acid concentration and proportion of DCA were significantly low in bile from Group C and decreased proportion of DCA and increased proportion of CDCA were found in bile from Group B. In both the gall-bladder and hepatic bile, proportion of LCA was not significantly different between any intergroups.(ABSTRACT TRUNCATED AT 250 WORDS)

The inverting and everting methods of biliobiliary anastomoses were compared histopathologically and electron microscopically. Epithelialization started on the 3rd postoperative day and occurred within 8 mm of the anastomosis with an earlier and more active epithelialization being seen in the proximal area than in the distal. A rapid decrease of the mucosal defect was seen for 3-5 days which slowed down thereafter, and closure was achieved by 30 days after the anastomosis in both groups. The everted group showed a smoother and more rapid healing process, quicker epithelialization, and less mucosal defects than the inverted group. In the chronic phase, the inverted group showed more papillary hyperplasia and more pronounced fibrosis of the wall. The stricture index, being the internal circumference of the common bile duct: proximal x 2/anastomosis site + duodenal x 100, of the everted group was 123.7% compared to 146.7% for the inverted group, but there was no statistical difference. There was no difference in the total area of crypts, representing the epithelialization, between the two groups. Anastomoses with proximal dilatation therefore healed more slowly than those without dilatation. These findings show everted anastomosis to be superior to inverted anastomosis and thus support the usefulness of T-tube drainage to prevent postoperative dilatation of the bile duct.

To investigate the relationship between biliary mucin and ductular stone formation, mucin was isolated from hepatic bile using gel filtration on Sepharose CL-4B. The bile was obtained from 14 patients with stones in various sites of the biliary tract. The hexose content in the excluded fraction was significantly higher in patients with intrahepatic ductular stones (68.7 +/- 20.5 micrograms/mL; mean +/- s.d.) than in those with gall-bladder stones or extrahepatic ductular stones (23.8 +/- 8.1 micrograms/mL, 33.3 +/- 9.5 micrograms/mL; P less than 0.05), suggesting a higher concentration of mucin in the bile of patients with intrahepatic ductular stones. Ion-exchange chromatography on DEAE-Sephacel showed that most mucin from each material was negatively charged and electrophoretic studies indicated that it was composed mainly of high molecular weight (greater than 10(6)), sulfated glycoprotein. These results suggested that the mucin content of hepatic bile might have an important relation to the development of intrahepatic ductular stones.

The aim of the present study was to analyse bile samples from cases with gallstones by high performance liquid chromatography according to the type of stones present, with special reference to the glucoside and xyloside conjugates of bilirubin, and to investigate their deconjugation. The composition of bilirubin conjugates in bile was similar between cholesterol and black pigment stones except that the total bilirubin concentration was about 5 times higher in black pigment stone cases with haemolysis. Unconjugated bilirubin was higher in brown pigment stone cases than in cholesterol stone cases, although total bilirubin concentration was lower in the former. In addition, in brown pigment stone cases, bile contained statistically less bilirubin diglucuronide and more bilirubin diglucoside and monoglucoside than in bile with cholesterol stones (P less than 0.05). Glucoside and xyloside conjugates are also major components, regardless of the types of gallstones present, accounting for as much as 18 to 25%. Incubation experiment revealed that bilirubin diglucuronide was more readily deconjugated than bilirubin diglucoside or bilirubin monoglucoside monoxyloside. Therefore, glucuronide conjugates were likely to be the main source of unconjugated bilirubin in the formation of pigment gallstones.

We examined 30 surgically removed gallbladders with ultrasound using a 10 MHz transducer in a water bath. The sonographic structure of gallbladder wall was imaged as 5 layers. The layer 3, a well-defined hyperechoic zone, was confirmed histologically as corresponding to the boundary between subserosa and muscularis. Since the spreading of gallbladder carcinoma beyond the muscle coat is an important prognostic indicator, high resolution images may well be used to advantage in assessing gallbladder wall penetration intraoperatively, enabling the surgeon to choose the proper operative procedure.

A clinicopathologic study of 40 cases of carcinoma of the gallbladder is presented. Twenty-six cases resected were assessed retrospectively with respect to the operative procedures employed and the results based on the pathologic findings from the resected specimens. The relationship between clinical features, macroscopic forms of tumor, histological types, liver invasion, and lymph node metastasis were investigated. Papillary, papillary infiltrative and nodular forms were classified as either papillary adenocarcinoma or well-differentiated tubular adenocarcinoma and invasion of the liver and lymph node metastasis were rare. Frequent lymph node metastasis was encountered in the nodular infiltrative form and invasion of the liver was frequently present in the infiltrative form. Invasion of the liver, lymph node metastasis, and the presence of gallstones were less frequent in papillary adenocarcinoma. In contrast, moderately-differentiated tubular adenocarcinoma frequently had lymph node metastasis. Invasion of the liver and lymph node metastasis were, however, present regardless of the histologic types and were more related to the extent of subserosal involvement present. A female preponderance was noted in poorly-differentiated adenocarcinoma. The main reasons for surgery being limited to exploratory laparotomy only or palliative procedures included carcinoma infiltration into the hepatoduodenal ligament, carcinoma extension to the neighboring structures, multiple liver metastases, peritoneal dissemination, large liver invasion, and multiple metastases to the paraaortic lymph nodes.

The incidence of hepatolithiasis is high throughout East Asia compared with the West, but the marked difference in the relative proportion of hepatolithiasis to all cholelithiasis cases exists even among countries of similar ethnic backgrounds. A retrospective study of cases was conducted in two areas in China with the aim of clarifying the presence of such regional difference in China itself. The relative proportion of hepatolithiasis was 21.2% in Shenyang, 9.2% in Beijing and 4.1% in Fukuoka, Japan. A significant difference in the location of stones was also found between Shenyang, Beijing and Fukuoka. Intra- and extrahepatic hepatolithiasis in all hepatolithiasis cases was 95% in Shenyang and 75% in Beijing. Involvement of both hepatic lobes was found in 73% in Shenyang and less than 60% in the other two, suggesting that hepatolithiasis of the old form or of an advanced stage still lingers in Shenyang. In conclusion, regional differences in the proportion and the type of hepatolithiasis exist in China itself, as well as in the Chinese population in Taiwan, Hong Kong and Singapore, as previously reported. The possible contribution of environmental factors to the occurrence of hepatolithiasis is again emphasized.

A high incidence of inflammation and carcinoma of the biliary tract in patients with anomalous pancreaticobiliary ductal junction has been well documented. The change in biliary phospholipids as a result of the reflux of pancreatic juice into the biliary tract through anomalous pancreaticobiliary ductal junction may be responsible for it. We developed a new method of analysis of phospholipid classes using aminopropyl Bond Elut cartridge for extraction and high-performance liquid chromatography for separation. Satisfactory recovery was achieved (i.e., more than 95% for both phosphatidylcholine and lysophosphatidylcholine). With this method, the bile of 11 patients with anomalous pancreaticobiliary ductal junction was examined. The concentration and proportion of lysophosphatidylcholine in bile were much higher in the presence of anomalous pancreaticobiliary ductal junction than in controls (3.44 +/- 1.50 mmol/L vs. 0.52 +/- 0.25 mmol/L, 60.0% +/- 31.0% vs. 2.3% +/- 1.4% in gallbladder bile; p less than 0.001). In contrast, the concentration of phosphatidylcholine and the sum of phosphatidylcholine and lysophosphatidylcholine in gallbladder bile significantly decreased (p less than 0.001), but in hepatic bile they did not. An inverse correlation was found between the proportion of lysophosphatidylcholine and phospholipid concentration in gallbladder bile. Phospholipase A2 and amylase activities in bile were markedly high. Increased total fatty acid concentration and proportion of unsaturated fatty acid in bile were found. Total bile acid concentration in gallbladder bile was significantly lower than in controls. These results suggest that a considerable amount of lysophosphatidylcholine, which is known to have a cytotoxic effect, isp reduced by phospholipase A2 in refluxing pancreatic juice, and an increased concentration of lysophosphatidylcholine gives rise to cell damage causing mucosal hyperplasia and metaplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum levels of 7 alpha-hydroxycholesterol and activities of hepatic microsomal cholesterol 7 alpha-hydroxylase in surgical patients were analyzed by capillary gas-liquid chromatography-selected ion monitoring technique using a new internal standard, 5 alpha-cholestane-3 beta, 7 beta-diol. We found that concentrations of 7 alpha-hydroxycholesterol obtained after alkaline hydrolysis were higher than those without alkaline hydrolysis, indicating that a considerable amount of 7 alpha-hydroxycholesterol in human serum is present in esterified form. Esterified 7 alpha-hydroxycholesterol could also be quantitatively hydrolyzed with cholesterol esterase, suggesting that fatty acid is bound at the 3 beta-position of the cholestenediol. The serum levels of esterified and free 7 alpha-hydroxycholesterol in patients with cholelithiasis were 198.0 +/- 90.3 and 48.3 +/- 19.8 pmol/ml (mean +/- SD), respectively, and were similar to those in patients without hepatobiliary diseases. After treatment with chenodeoxycholic acid (300 mg per day) for 7 to 10 days, esterified and free 7 alpha-hydroxycholesterol levels decreased to 64.9 +/- 33.6 and 20.5 +/- 11.1 pmol/ml, respectively. Activity of cholesterol 7 alpha-hydroxylase was also inhibited. Treatment with ursodeoxycholic acid (600 mg per day) for 7 to 10 days had no inhibitory effect on serum 7 alpha-hydroxycholesterol levels and the enzyme activity. In all groups, high correlations were found between the activity of cholesterol 7 alpha-hydroxylase and serum levels of 7 alpha-hydroxycholesterol: free (r = 0.71, n = 38, P less than 0.001); esterified (r = 0.87, n = 38, P less than 0.001); total (r = 0.87, n = 38, P less than 0.001). Esterified and total 7 alpha-hydroxycholesterol was more highly correlated with the enzyme activity than the free form. We conclude that a significant amount of 3 beta-acyl esters of 7 alpha-hydroxycholesterol is present in human serum and that serum levels of esterified and/or total 7 alpha-hydroxycholesterol are likely to reflect the activity of hepatic cholesterol 7 alpha-hydroxylase and thus the amount of primary bile acids synthesized in the liver.

Unscheduled DNA synthesis (UDS) induced by treatment with 20-methylcholanthrene (MCA) and N-nitrosodimethylamine (DMN) was measured in cultured human gallbladder epithelial cells. MCA induced UDS very efficiently, while DMN was far less effective than MCA. Addition of rat S9 mixture did not affect the amount of UDS by the chemicals. Differences between the present results in human cells and our previous findings in bovine cells could be due to species and tissue specificity.

A reliable method for the simultaneous determination of keto and non-keto bile acids in human serum was developed. Carbonyl substituents of bile acid ethyl esters were converted into methyloxime and hydroxyl substituents into dimethylethylsilyl ethers and the products were analysed directly by capillary gas chromatography with selected ion monitoring using [2H4]chenodeoxycholic and [2H4]3 alpha-hydroxy-7-oxo-5 beta-cholanoic acids as internal standards. The bile acid peaks on the selected ion chromatogram were separated without interference from endogenous substances present in serum. Recoveries of individual keto bile acids added to serum range from 74.4 to 94.7% with a mean of 87.1%. Eight kinds of keto bile acids not previously found in sera of normal subjects, namely 3-oxo-, 3-oxo-7 alpha-hydroxy-, 3-oxo-12 alpha-hydroxy-, 3 alpha-hydroxy-7-oxo, 3 alpha-hydroxy-12-oxo-, 3-oxo-7 alpha,12 alpha-dihydroxy-, 3 alpha,7 alpha-dihydroxy-12-oxo- and 3 alpha,12 alpha-dihydroxy-7-oxo-5 beta-cholanoic acids were identified and quantified. The total concentration of keto bile acids was found to be 0.16 +/- 0.08 nmol/ml and constituted 2.9 +/- 1.5% of that of the usual non-keto bile acids in peripheral venous serum.

The effects of dietary cholesterol and cholestyramine on pancreatic carcinogenesis initiated with N-nitrosobis(2-oxopropyl)amine (BOP) were investigated in 120 female Syrian golden hamsters. BOP (70 mg/kg body weight) was injected s.c. once at the beginning of the experiment. Starting 2 weeks later, the animals were then maintained on basal diet or diets containing either 0.5% cholesterol or 1% cholestyramine for a further 16 weeks. All surviving hamsters were killed at week 18, and the pancreas tissues examined histologically. The incidences of pancreatic carcinomas in hamsters fed cholesterol and the cholestyramine supplement were 40.0 and 30.0% respectively; in both cases significantly higher than the 6.9% incidence in the basal diet group. Cholesterol contents of the serum, pancreas and liver were significantly increased by cholesterol feeding and significantly decreased by the cholestyramine diet. The cholesterol diet also significantly increased pancreatic protein and DNA contents, and the concentration of total bile acids and the level of lithocholic acid in gallbladder bile. The cholestyramine diet significantly increased total pancreatic DNA and protein contents, and pancreatic weight. The results thus indicated that both dietary cholesterol and cholestyramine can enhance BOP-initiated pancreatic carcinogenesis in hamsters.

We have previously isolated from 13 cholesterol gallstones a low molecular weight acidic bili-protein that inhibited the precipitation of calcium carbonate in vitro. We now report the isolation of a similar protein from seven black pigment gallstones. Cholesterol was removed from the stones by Soxhlet apparatus with methyl t-butyl ether, and bile acids were extracted with methanol. The protein was purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis after demineralization of the stones with ethylenediaminetetraacetate. Structural and functional properties of the protein from the black stones that were similar to the protein from the cholesterol stones included the following: (a) an apparent molecular weight of about 5 kD; (b) a high content of acidic (19.8%) and hydrophobic (50.1%) amino acids with a low content of basic residues (8.4%) and little sulfide-containing amino acids (1.9%); (c) an inhibitory effect on both the initiation and growth of calcium carbonate crystals in vitro; and (d) very tight (possibly covalent) binding of a diazo-positive yellow pigment, presumably bilirubin, with maximum spectral absorbance at 410 nm. The structural and functional similarities of these bili-proteins from black pigment and cholesterol gallstones and their striking effects on calcium carbonate precipitation in vitro suggest that they play a common role in the regulation of precipitation of calcium salts during the formation of both types of gallstones. (HEPATOLOGY 1992;15:1079–1085).

The effects of treatment with chenodeoxycholic acid (CDCA) or ursodeoxycholic acid (UDCA) on nucleation time, biliary lipid concentration, and vesicular lipid composition were studied. Gallbladder bile was collected at the time of surgery from 33 cholesterol gallstone patients who were divided into three groups: 16 untreated, 9 pretreated with CDCA (400 mg/day), and 8 pretreated with UDCA (600 mg/day) for 1-3 weeks before surgery. Control bile samples were also collected from nine patients without cholelithiasis. Nucleation time was prolonged significantly in both CDCA- and UDCA-treated groups [12.6 +/- 8.5 (SD) and 21.0 +/- 0 days, respectively] compared with the untreated gallstone group (3.3 +/- 3.2 days). Both treatments significantly decreased the proportion and concentration of both cholesterol and phospholipids present in the vesicular phase. Treatment with UDCA decreased the cholesterol saturation index more than did CDCA at the dose used in this study. In the CDCA-treated group, patients without much change in cholesterol saturation index (greater than 1.0) showed a prolongation of the nucleation time with a significant decrease in vesicular cholesterol concentration, indicating a shift of cholesterol from vesicles to micelles. UDCA-treated patients and CDCA-treated patients with decreased cholesterol saturation index (less than 1.0) showed a greater effect. The authors conclude that UDCA prolongs the nucleation time mainly by decreasing the cholesterol saturation index, whereas CDCA does so by the dual effect of lowering the cholesterol saturation index and shifting cholesterol from vesicles to micelles.

This study examined the hypothesis that filtration through the MW 300 kD cut-off membrane (XM-300) may influence factors affecting the cholesterol nucleation time (the appearance time of cholesterol monohydrate crystal). Differences in biliary lipids, biliary protein, mucous glycoprotein and vesicular composition were examined before and after the XM-300 filtration in control and cholesterol gallstone patients. The nucleation time in the cholesterol gallstone patients was significantly faster than that in the control patients. However, the nucleation time in the gallstone patients was significantly prolonged following the XM-300 filtration resulting in a similar value to that of the control patients. No differences in concentrations of total protein, mucous glycoprotein or lipids composition were observed. The nucleation time did not correlate with the total lipid concentration, the concentrations of biliary mucous glycoprotein or total protein. The prolonged nucleation time of gallstone bile by the XM-300 filtration was primarily ascribed to the removal of vesicles, which was confirmed by gel chromatography. It was concluded that vesicles greater than MW 300 kD are primarily responsible for the rapid nucleation time.

Bile acid metabolism after two-thirds partial hepatectomy (PH) in rat was studied. Bile acid kinetics (i.e. synthesis rate and pool size) were determined by wash out method combined with gas liquid chromatography, and serum bile acids by gas liquid chromatography-mass spectrometry. Serum bile acid concentration was highest on the third day after PH, as the liver regeneration progressed but it gradually decreased with increasing cholic acid (CA)/chenodeoxycholic acid (CDCA), reflecting impaired hepatic uptake of bile acids and/or cholestasis during the early post-hepatectomy period. Predominance of CA in bile acid synthesis, pool, and biliary secretion was also found. On the third day after PH, liver weight recovered to 66% of the control value, but enhancement of bile acid synthesis was not observed. Consequently, pool size remained at 50% of control. On the seventh day, synthesis of bile acid, especially of CA, was enhanced and pool size and liver weight returned to 68 and 72% of the respective control values. Bile acid synthesis was returned to the control value on the fourteenth day with concomitant restoration of liver weight and bile acid pool size. These changes in bile acid kinetics parallel the events during hepatic regeneration after PH.

The profiling of eicosanoids, including prostaglandin E2 (PGE2), prostaglandin F2 alpha (PGF2 alpha), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), thromboxane B2 (TXB2) and leukotriene B4 (LTB4), in dog and human gall bladders was carried out by a combination of an effective and convenient clean-up procedure and gas chromatography with selected-ion monitoring. The clean-up procedure was based on the stepwise elution of their methyl ester derivatives from a silica gel column with n-hexane-ethyl acetate and ethyl acetate-methanol in various ratios. The LTB4 methyl ester was eluted with an n-hexane-ethyl acetate (2:1, v/v) fraction because LTB4 is more lipophilic than the other eicosanoids. The present method permitted the quantitation of trace amounts of eicosanoids, including LTB4, present in tissues in the order of pg/mg of protein, without interference from other endogenous substances. In experimental acalculous cholecystitis produced in dog, the levels of eicosanoids (except LTB4) were significantly changed. Of these eicosanoids, the level of 6-keto-PGF1 alpha was significantly higher in the seromuscular layer and correlated with the observed severe morphological changes. In human chronic cholecystitis with gallstones, the level of 6-keto-PGF1 alpha in the mucosal layer was significantly higher than that in the seromuscular layer. These data suggest that prostaglandin I2 may play an important pathophysiological role in the course of cholecystitis.

The effect of truncal vagotomy (TV) on pancreatic carcinogenesis initiated with N-nitrosobis(2-oxopropyl)amine (BOP) was investigated in 81 female Syrian golden hamsters. The animals were divided into four groups according to the treatment, groups 1 and 2 serving as non-initiated controls receiving a single s.c. injection of 0.9% NaCl followed by either a sham operation or TV respectively, at week 2. Groups 3 and 4 were given a single s.c. injection of 70 mg/kg body wt of BOP before the sham operation or TV. All hamsters were killed at week 24, and the pancreas, liver and gall bladder tissues were examined histologically. While TV itself caused no significant change in pancreatic weight, the incidence of pancreatic carcinomas in hamsters from group 4 was 48.4%, significantly higher than the 16.7% evident in hamsters from group 3 (P less than 0.05). GLC analysis of the bile acid composition of gall bladder bile from hamsters not receiving carcinogen 1 and 4 months after TV revealed significantly decreased secondary bile acids. The results thus indicated that changes in bile acid composition may be involved in enhancement of BOP-initiated pancreatic carcinogenesis in hamsters by TV.

Among 26 patients with gallbladder carcinoma, 11 resected cases were studied retrospectively with respect to the preoperative imaging techniques and the operative procedures employed. Ultrasonography, endoscopic ultrasonography, and computed tomography were valuable in detecting gallbladder carcinoma preoperatively while direct cholangiography and angiography were helpful in designing operative strategy. The ultrasonographic findings were: a polypoid (fungating) mass protruding into the gallbladder lumen in 45%, abnormal thickening of the gallbladder wall in 18%, and a mass in the gallbladder invading adjacent organs in 18% of cases. Approximately 80% was diagnosed preoperatively by ultrasonography. Correct diagnosis was made by computed tomography in 60% of the cases. Endoscopic ultrasonography offered valuable information on the depth of tumor invasion. Direct cholangiography and angiography were useful in assessing the extent of the tumor spread. Recent advances in hepatobiliary imaging techniques, especially ultrasonography and computed tomography, contributed greatly to the detection and evaluation of gallbladder carcinoma at an early and resectable stage.

In order to investigate the effects of bile acid feeding on hepatic microsomal deoxycholate 7 alpha-hydroxylase activity, three different bile acids were administered (0.2% w/w in chow) to hamsters for two weeks. Deoxycholate 7 alpha-hydroxylase activity was increased markedly by feeding of cholic acid (CA) and slightly by deoxycholic acid (DCA). Chenodeoxycholic acid (CDCA) had little effect on the enzyme activity. Feeding each of the bile acids significantly inhibited the activity of cholesterol 7 alpha-hydroxylase in the order CDCA greater than or equal to DCA greater than CA. There was no correlation between deoxycholate 7 alpha-hydroxylase activity and cholesterol 7 alpha-hydroxylase activity. It is concluded that the activity of deoxycholate 7 alpha-hydroxylase is up-regulated by feeding DCA and CA and that the mechanism seems to be different from that of cholesterol 7 alpha-hydroxylase. The increased activity of hepatic deoxycholate 7 alpha-hydroxylase by CA and DCA should be beneficial in minimizing the toxic effects of DCA in the hamster.

The bile acids in brown pigment stones and gallbladder bile were fractionated into free acids, glycine and taurine conjugates, and sulfates using diethylaminohydroxypropyl Sephadex LH-20 (DEAP-LH-20) column chromatography and were quantitated by gas chromatography. Twenty-eight cases of brown pigment stones were studied and divided into two groups: those with and those without bacteria possessing bile acid-deconjugating activity. In the former, free bile acid amounted to 62 +/- 34% of the total bile acid, while in the latter, only 0.1% of total bile acid was free bile acid. The fraction of total bile acid made up of free bile acids was found to be consistently higher in brown pigment stones than in the corresponding bile, irrespective of the presence or absence of biliary infection. Free bile acid is present in negligible amounts in normal bile. Total bile acid concentration in the bile of patients with brown pigment stones was significantly less than that of controls (13 vs 50 mg/ml). Biliary infection is almost always present in cases with brown pigment stones. These findings suggest that bacterial infection is present at the initiation of brown pigment stone formation as well as during the period of ensuing stone growth.

Monohydroxy bile acids in liver tissue may be of importance because of their hepatotoxicity and strong cholestatic effects. Recently, the existence of lithocholate in liver tissue in two forms was suggested by Nair et al. (Lipids. 1977. 12: 922-929) i.e., either in free form or as so-called tissue-bound lithocholate released exclusively by cholylglycine hydrolase treatment. The presence of the latter aroused much interest in relation to its hepatotoxicity and possible role in tumor induction. In the present investigation lithocholyl-epsilon-L-lysine, proposed as the predominant tissue-bound bile acid, was synthesized and its metabolic behavior was tested. Lithocholyl-epsilon-lysine was not deconjugated by cholylglycine hydrolase treatment but only by alkaline hydrolysis. Bile acids in seven cirrhotic and three noncirrhotic liver samples were extracted with 95% ethanol-0.1% ammonium hydroxide. The bile acids in the extract and residue were quantified by glass capillary gas-liquid chromatography using selected ion monitoring. The presence of so-called tissue-bound lithocholate could not be substantiated in either cirrhotic or noncirrhotic liver tissues. Nearly complete extraction of lithocholate was achieved by the use of organic solvent alone. Therefore, tissue-bound lithocholate, if it exists at all, may be attached to tissue by a physical linkage which can be disrupted by the use of conventional organic solvent.

Changes in intestinal absorptive capacity for water and electrolytes were investigated in dogs after total colectomy and mucosal proctectomy reconstructed with interposing jejunum into the anorectal area. Rate of abosorption of water and sodium from the ileum increased significantly at 29 weeks postoperatively. The absorption of water, sodium, and chloride in the jejunum was significantly higher than in the neorectum. The net secretion rate of potassium increased significantly in the ileum and neorectum. The authors suggest that intestinal adaptation achieved after proctocolectomy is enhanced in the jejunum and ileum rather than in the neorectum.

Duodenoscopic manometry of the pancreatic duct (PD) and common bile duct (CBD) using a microtransducer catheter was distinct advantages over infusion manometry, giving absolute values of in situ intraluminal pressure. Microtransducer manometry was performed without medication in 49 patients with gallbladder stones (10), common bile duct stones (24), hepatic duct stones (6) and common bile duct dilatation (9), and was successful in 42 (86%) for PD and 36 (73%) for CBD. Ductal pressures showed respiration-synchronized biphasic variations superimposed by the arterial pulsation effect. Considerable postural change of the pressure values suggested that the recording posture should be predetermined. The PD-to-duodenum pressure gradient was higher than the CBD-to-duodenum gradient in most cases. Both were lower than those obtained previously by infusion methods. No significant differences were found in pressure profiles of the four disease groups. Endoscopic sphincterotomy significantly reduced not only CBD pressure but also PD pressure.

One thousand thirty-eight patients with primary gastric cancer treated operatively during the past 20 year period were described. The results were compared with those in the period from 1904 to 1927 to illustrate the changes which have taken place in diagnosis and treatment of gastric cancer during the past 76 years in Japan. All data was from the Department of Surgery I, Kyushu University. After half a century of status quo, improvement started around the mid 1960s, which coincided with the introduction of newer, improved procedures for diagnosis of gastric cancer, such as double contrast roentgenography of the stomach, endoscopic observation, and biopsy of suspected lesions. A mass population survey for the presence of gastric cancer in persons of approximately 40 years of age has led to an ever increasing number of early gastric cancer cases. The emergence of early simulating advanced cancer with relatively good prognosis and the changing state of the Borrmann type of advanced cancer have contributed to the improvement. The present study demonstrates once again the importance of early detection of gastric cancer for an increased chance of cure, since conventional lymph node dissection seems to be unneccessary, and of even earlier detection of gastric tumors of less than 1 cm in diameter for the best chance of complete cure in the latter.

Dehydrocholic acid was administered intravenously to a patient and to rats, and its metabolites excreted into bile were studied by gas chromatography-mass spectrometry as ethyl ester-dimethylethylsilyl ether-methyloxime derivatives. Two 7β-hydroxy metabolites were identified : 3α, 7β-dihydroxy-12-oxo-5β-cholanoic acid in both human and rat bile, and 3α, 7β, 12α-trihydroxy-5β-cholanoic acid in rat bile. This indicates that human and rat livers are able to reduce the 7-oxo group in dehydrocholic acid to both 7α-and 7β-hydroxy forms.

Nucleation time (Cholesterol monohydrate crystallization) in bile has been introduced to distinguish patients with cholesterol gallstone from nomal human beings. To test the hypothesis that cholesterol monomer activity correlates with nucleation time, we have examined the relationship between nucleation time, apparent cholesterol monomer activity and cholesterol monomer concentration in the aqueous phase using model biles containing bile salt, phospholipid and cholesterol. Apparent cholesterol monomer activity was inversely well correlated with nucleation time in the dilution study ( r = −0.98) and in the cholesterol saturation study ( r = −0.88) but positively with cholesterol monomer concentration in the aqueous phase in the dilution study ( r = 0.72) and in the cholesterol saturation study ( r = 0.93). The results indicate that cholesterol monomer activity reflects the nucleation time in model bile system.

Sphincter of Oddi phasic motor activity and common bile duct pressure were investigated in controls (seven patients) and in patients with gall-bladder stones (five patients), common bile duct stones (15 patients), or intrahepatic stones (13 patients). There were no significant differences in amplitude and frequency of the phasic activity or the common bile duct pressure between the controls and disease groups. Basal pressure of the phasic contraction, however, was significantly lower in patients with common bile duct or intrahepatic stones than in the controls or gall-bladder stone group. The administration of morphine, known to cause spasm of the sphincter of Oddi, increased the basal pressure and frequency of the phasic waves in all groups, while the amplitude remained unchanged. Response to morphine in patients with common bile duct or intrahepatic stones was similar to the controls. However, the basal pressure in these latter groups was lower than in the controls, even after stimulation by morphine. The high incidence of bacterial growth in bile from these patients hitherto reported may be attributable to ascending infection possibly resulting from the low basal pressure of the sphincter of Oddi.

Urinary excretion of bile alcohols and bile acids in patients with complete extrahepatic cholestasis before and after the release by external biliary drainage was studied. Following extraction, isolation, and hydrolysis, bile alcohols were determined by capillary gas-liquid chromatography-mass spectrometry as dimethylethylsilyl derivatives. During cholestasis, 8.89 mumol/day of bile alcohol and 140.4 mumol/day of bile acid were excreted in urine. The amount of bile alcohol excreted was 6.1% of that of bile acid. Positive correlation between excretion of bile alcohols and bile acids was observed. The major bile alcohols excreted were also present in urine from healthy individuals but in much smaller amounts. After the release of extrahepatic cholestasis, urinary excretion of bile acid decreased rapidly, but that of bile alcohol decreased only gradually. The latter often increased again and remained high. The results indicate that the increased excretion of bile alcohols in complete extrahepatic cholestasis may reflect the expansion of a normally existing pathway of bile alcohol synthesis and excretion leading to the modification of bile alcohols for their efficient urinary elimination. It is also suggested that the rate of synthesis of bile alcohols is determined partly by the size of the substrate pool available.

Bile acid analysis of mild and severe cirrhotic liver showed that with the advancement of cirrhosis the concentration of chenodeoxycholic acid in liver tissue becomes higher, resulting in the lower ratio of cholic to chenodeoxycholic acid probably due to the progressive alteration of cholic and chenodeoxycholic acid synthesis with the advancement of liver cirrhosis. Bile acid analysis of paired liver and bile of severe cirrhosis showed that the ratio of cholic to chenodeoxycholic acid in liver was lower than that in bile or even with that in bile. This can be explained by postulating the impaired hepatic secretion of bile acids, especially chenodeoxycholic acid. The impaired secretion together with the relatively well preserved chenodeoxycholic acid synthesis results in the accumulation of chenodeoxycholic acid in liver tissue with cirrhosis.

Changes of bile composition in gallbladder bile and serum chemistries were investigated in nine dogs after proctocolectomy and ileoanal anastomosis, using an interposed jejunal segment as a neorectum. A significant decrease in cholic acid and an increase in deoxycholic acid were observed 24 weeks after the operation, although there was no significant change of total bile acid and phospholipids in the bile. Concentration of cholesterol and cholesterol saturation index in bile increased after total proctocolectomy. Although serum triglyceride levels became lower compared with the nontreated control group, concentrations of total protein, serum cholesterol, total lipids, blood sugar, and electrolytes showed no significant difference. This operative procedure apparently did not impair intestinal absorption of bile acid, but did increase the biliary cholesterol saturation index.

This investigation was undertaken to clarify the mechanism of micropore-formation in WC-Co alloys, from the viewpoint that pores would be resulted from coarse Co particles which pre-existed in (WC+Co) mixture. High carbon two-phase WC-Co alloys with about 10 and 20% Co were in vacuum sintered under various conditions, using usually ball-milled mixtures in which some amounts of coarse Co particles were intermixed for a model-experiment, or from which pre-existed coarse Co particles were removed or not.Results obtained were as follows:1) In case of the low temperature sintering including solid-phase sintering, coarse Co particles in the mixture became Co pools and changed into pores as sintering time increased. In case of the high temperature sintering, those Co pools tended to change rapidly in pores, when the Co content in the alloy was low. 2) Thus, the structural defects such as Co pools and pores were confirmed to be closely related to each other. Pores above mentioned didn't develope in the alloys, for instance, with high Co contents because of the facility of viscous flow of the matrix. 3) The above results in model-experiments were considered to be available also in the usual WC-10% Co alloy. Because, the removal of coarse Co particles which pre-existed in the ball-milled mixture resulted in disappearence of the pores, the dimensions of which corresponded to those of removed Co particles, leading to a sharp increase in the strength of alloys.

In a recent publication, we reported that deoxycholic acid is 7 alpha-hydroxylated to yield glycocholate or taurocholate in vivo in the hamster (1987. Kuroki et al. Hepatology. 7: 229-234). In order to explore the possibility that amidation of free deoxycholic acid precedes the 7 alpha-hydroxylation, we assayed 7 alpha-hydroxylase activities of free and conjugated deoxycholates in vitro. 7 alpha-Hydroxylase activities of glycodeoxycholate and taurodeoxycholate were 720 +/- 132 and 640 +/- 160 pmol/mg.min-1, respectively. Activity of 7 alpha-hydroxylation of free deoxycholate was very low (60 +/- 20 pmol/mg.min-1). After treatment with phenobarbital in a dose of 100 mg/kg per day for 6 days, 7 alpha-hydroxylase activities of conjugated deoxycholates were decreased significantly (40%, P less than 0.01, n = 8), whereas that of free deoxycholate was not significantly changed. In the rat, 7 alpha-hydroxylase activities of conjugated deoxycholates were induced significantly (45% increase, P less than 0.05, n = 5) by phenobarbital treatment in sharp contrast to the hamster. There were significant correlations between the 7 alpha-hydroxylase activity of taurodeoxycholate and that of glycodeoxycholate both in the hamster and in the rat (hamsters: n = 16, r = 0.98, P less than 0.01; rats: n = 10, r = 0.82, P less than 0.01). These studies suggested that deoxycholic acid is 7 alpha-hydroxylated after amidation with glycine or taurine in vivo and that the same enzyme may well catalyze the 7 alpha-hydroxylation of glycodeoxycholate and taurodeoxycholate in the hamster.(ABSTRACT TRUNCATED AT 250 WORDS)

It is well known that the strength of WC-Co low carbon cemented carbide decreases with precipitation of Co3W, when the alloy is annealed at about 800°C. The present study was undertaken to reveal the reason of the strength decrease, in relation to the structural defects which would act as a fracture source. The WC-10 and 20% Co low carbon two-phase alloys (mean grain size of carbide, about 1.2 μ) were annealed at 800°C for up to 15 hr, and used as specimens. After the transverserupture test had been performed according to the Japanese Industrial Standard, measurements of the sort, dimension and location of the defects were made on the fracture surface.Results obtained were as follows. (1) It was shown that the sort, dimension distribution and average dimension of the defects as a fracture source didn't vary by the precipitation treatment, that is, neither the development of new defects nor the growth of the pre-xisting defects took place. (2) The strength of the sound matrix (σ0) decreased by the treatment. Thus, it became clear that the strength decrease was resulted from the decrease of σ0 only. (3) However, further investigation is needed to make clear the machanism of the decrease of σ0.

A total of 38 patients with middle and distal thirds bile duct cancer treated in our department during a period of 20 years were reviewed and analyzed in respect to the type of operative procedures employed and the long-term results. The purpose of the study was to evaluate the impact of various recently available modalities for diagnosis and treatment on the prognosis of patients with these malignancies.

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.

A method of microquantitative determination of bile acid in 5–30 mg of human liver tissue was developed. Bile acids were converted to their ethyl ester dimethylethylsilyl ether derivatives and analyzed by capillary gas chromatography-selected ion monitoring, using [2H4]-lithocholic (LCA), [2H5]deoxycholic (DCA), [2H4]chenodeoxycholic (CDCA), [2H4]-ursodeoxycholic (UDCA), and [2H3]cholic (CA) acids as internal standards. Precision and reproducibility of the present method were tested using surgically obtained liver specimens. The results were statistically analyzed according to one-way layout and the orthogonal polynomial equation. Bile acids except LCA were determined with 2.3 to 11.4% of the coefficient of variation. Recoveries of conjugated bile acids ranged from 72.2 to 96.0% with a mean of 84.3%. The amount of bile acids present in histologically normal liver specimens (n = 10) was found to be 29.56 ± 8.62 μg/g liver. The relative compositions (%) of CDCA and CA were 38.8 ± 8.9 and 41.1 ± 11.0, respectively.

The WC-(10, 20, 30)%Co alloys were minutely vacuum-sintered, so that the defect acting as a fracture source in each alloy might almost consist of coarse WC grains smaller than about 20μ. Two sorts of cooling rates after sintering, about 15°and 0.5°C/min, were adopted in the temperature range from 1360°to 1300°C for 10%Co alloy, 1350°to 1300°C for 20%Co alloy and 1340°to 1300°C for 30%Co alloy, respectively. Two sorts of specimens different in cooling rates in each alloy were ground with a diamond wheel, and transverse-ruptured according to JIS. After the test, the dimension (2a) and location of coarse WC grains as a fracture source were measured on the ruptured surface.The results obtained were as follows: 1) The average transverse-rupture strength (σm), and also the external stress (σd) corresponding to σm were always higher in rapidly cooled specimen (R-specimen) than in slowly cooled specimen (S-specimen). 2) The 2a of R-specimen was generally smaller than that of S-specimen, showing that of an anomalous grain growth of WC also occurred during the time of cooling specimens, when the cooling rate was decreased. 3) The distance from the tension surface to the defect (Δt) of R-specimen was larger in 10%Co alloy than that of S-specimen. However, the difference in Δt was hardly observed in (20, 30)%Co alloys. It was found in a slowly cooled 30% Co alloy that the location of the defect was always on the domain boundary of binder phase. 4) The increase in Δm and σd due to increasing cooling rate was considered to be affected mainly by the increase in the strength of WC-Co matrix including that of the domain boundary, and partially by the changes in 2a and Δt.

The transverse-rupture strength of conventional WC-10%Co high carbon two-phase alloys (mean grain size is about 1.4μ) was measured as a function of width (w) or thickness (t) of the specimen. Then, the observed strength was examined in relation to the structural defects which appeared on the fracture surfaces as a fracture source.Results obtained were as follows. (1) The increase of w or t resulted in the decrease of strength, showing the so-called volume effect. (2) The strength decrease was accompanied by the increase of the average defect (or source) dimension, and by that of the average distances from the tension surface and also from the span center to the defect. It was also accompanied by the change in the sort of defects. Thus, the volume effect as above was considered not to be simply related to usually cited risky volume, but to be related to the volume containing a defect which actually acts as a fracture source. (3) The effect of specimen-volume on the strength of conventional cemented carbide could not be satisfactorily explained by Weibull's equation.

The preparation and antigenic properties of 3β-hydroxy-5-cholen-24-oyl-glycine-bovine serum albumin (BSA) conjugate in which the hapten is linked to the carrier protein through an (O-carboxymethyl) oxime bridge at the C-19 position on the steroid portion are described. Antibody raised against the antigen in rabbits possessed high titer and specificity to 3β-hydroxy-5-cholen-24-oyl-glycine, exhibiting no significant cross-reactions with various bile acids.