Reversibility of hepatic mitochondrial damage in rats with long-term cholestasis

BACKGROUND/AIMS: Long-term bile duct ligation in rats is associated with secondary biliary cirrhosis and metabolic alterations, e.g. mitochondrial dysfunction. We performed the current studies to characterize the reversibility of hepatic mitochondrial dysfunction after reversing biliary obstruction by Roux-en-Y anastomosis. METHODS: Rats were studied after 4 weeks of bile duct ligation, and after 5 or 14 days of reanastomosis. Control rats were pair-fed to treated rats and all rats were studied after starvation for 24 h. Mitochondria were isolated by differential centrifugation and enzyme activities determined by spectrophotometric methods. RESULTS: In comparison to controls, plasma beta-hydroxybutyrate concentrations were decreased in bile duct ligated rats (200+/-70 vs. 790+/-200 micromol/l) and remained decreased after relief of biliary obstruction. In contrast, plasma free fatty acids were not different between controls and treated rats. Oxidative metabolism of L-glutamate, succinate and duroquinol was decreased in liver mitochondria from bile duct ligated rats. After relief of biliary obstruction, the metabolism of L-glutamate and duroquinol normalized quickly, whereas succinate metabolism remained impaired. Similar results were obtained for the mitochondrial oxidases in disrupted mitochondria. The activities of complex I, II, III and V of the respiratory chain were reduced in bile duct ligated rats. After relief of biliary obstruction, complex I and III normalized quickly, whereas complex II and V remained impaired. Oxidative metabolism of long-chain fatty acids by isolated liver mitochondria was decreased in bile duct ligated rats and did not recover after relief of biliary obstruction. CONCLUSIONS: Long-term cholestasis in the rat is associated with a decrease in specific functions of liver mitochondria which recover only partially after Roux-en-Y anastomosis. The persistence of decreased mitochondrial fatty acid metabolism cannot be explained by impaired activity of the respiratory chain, but is more likely due to alterations in mitochondrial beta-oxidation.     

Hematopoietic bone marrow hyperplasia: correlation of spinal MR findings, hematologic parameters, and bone mineral density in endurance athletes

To clarify the effects of obstructive jaundice on the liver, sequential changes of hepatic energy charge, the concentrations of adenine nucleotides and malondialdehyde, DNA synthesis rate, and histology of the liver were examined on the day before and Days 1, 2, 4, 7, and 14 after biliary obstruction in rats and compared with those of sham-operated controls. Foci of necrotic hepatocytes were present on Days 1 and 2 and mitoses of the hepatocytes were frequently observed with a peak on Day 2 in the jaundiced liver. Marked proliferation of bile ductules were subsequently observed on Days 7 and 14, resembling biliary cirrhosis. The DNA synthesis rate was significantly activated after bile duct obstruction with its peak on Day 2, more than nine times higher than the control value and returned to the control level on Day 14. Hepatic ATP concentration and energy charge gradually declined with prolonged jaundice and significantly lower levels persisted after Day 7 compared with the controls. The malondialdehyde level in the jaundiced liver gradually increased and became significantly higher on Day 14. We conclude that obstructive jaundice decreases hepatic energy charge and increases the lipoperoxide level. In the initial stage of obstructive jaundice, the hepatocytes proliferate associated with activated DNA synthesis probably to compensate hepatic damage; however, prolonged obstructive jaundice induces functional hepatic injury possibly necessitating biliary drainage.     

Dissociation between anxiolytic and hypomnestic effects for combined extracts of zingiber officinale and ginkgo biloba, as opposed to diazepam

Extrahepatic biliary atresia is a severe neonatal liver disease resulting from a sclerosing cholangiopathy of unknown etiology. Although biliary obstruction may be surgically corrected by a "Kasai" hepatoportoenterostomy, most patients still develop progressive hepatic fibrosis, although the source of increased collagen deposition is unclear. This study examined the role of hepatic stellate cells (HSCs) and assessed the source of transforming growth factor-beta (TGF-beta) production in hepatic fibrogenesis in patients with biliary atresia. Liver biopsies from 18 biliary atresia patients (including 5 pre- and post-Kasai) were subjected to immunohistochemistry for alpha-smooth muscle actin and in situ hybridization for either procollagen alpha1 (I) mRNA or TGF-beta1 mRNA. Sections were also subjected to immunohistochemistry for active TGF-beta1 protein. The role of Kupffer cells in TGF-beta1 production was assessed by immunohistochemistry for CD68. Procollagen alpha1 (I) mRNA was colocalized to alpha-smooth muscle actin-positive HSCs within the region of increased collagen protein deposition in fibrotic septa and surrounding hyperplastic bile ducts. The number of activated HSCs was decreased in only one post-Kasai biopsy. TGF-beta1 mRNA expression was demonstrated in bile duct epithelial cells and activated HSCs and in hepatocytes in close proximity to fibrotic septa. Active TGF-beta1 protein was demonstrated in bile duct epithelial cells and activated HSCs. This study provides evidence that activated HSCs are responsible for increased collagen production in patients with biliary atresia and therefore play a definitive role in the fibrogenic process. We have also shown that bile duct epithelial cells, HSCs, and hepatocytes are all involved in the production of the profibrogenic cytokine, TGF-beta1.     

Thyroid hormone is required for dietary fish oil to induce hypersecretion of biliary cholesterol in the rat

In the rat, both fish oil diet and thyroid hormone replacement are reported to augment bile cholesterol secretion out of proportion to bile flow or secretion of other bile lipids. We sought common mechanisms for these effects and evaluated the role of phospholipid fatty acid composition in the process. Methimazole-treated hypothyroid rats were fed low-fat chow or chow supplemented with 10% corn oil or fish oil, and were studied before and after thyroid hormone treatment. Serum, hepatic, and bile lipids were measured, phospholipid fatty acid composition determined, and hepatic 3-hydroxy-3-methylglutaryl CoA reductase activity assayed. Fish oil diet stimulated cholesterol secretion into bile only after thyroid hormone was given, and this action was synergistic with that of thyroid hormone. Reduced serum cholesterol in fish oil-treated rats was associated with increased biliary cholesterol secretion and diminished hepatic cholesterol content. This suggests that augmented biliary cholesterol secretion may contribute to the fish oil-induced reduction of serum cholesterol. No definite relationship between hepatic or biliary phospholipid fatty acid composition and biliary secretion was apparent, although high bile cholesterol secretion was associated with a low percentage of hepatic and bile phospholipid linoleic acid.     

Hepatic lobar atrophy: association with ipsilateral portal vein obstruction

OBJECTIVE: This study was performed to evaluate the association between hepatic lobar atrophy, bile duct obstruction, and portal vein obstruction. MATERIALS AND METHODS: Thirty cases of hepatic lobar atrophy identified on angiography with CT during arterial portography from August 1992 to March 1995 were retrospectively reviewed by two independent observers. Cases were evaluated for vascular patency and bile duct obstruction. Malignant diagnoses were present in 28 of 30 patients. RESULTS: Twenty-two patients (73%) had atrophy in the left lobe and eight patients (27%) had right lobar atrophy. Portal vein obstruction was unilateral and confined to the atrophic lobe in 26 patients (87%). In contrast, bile duct obstruction was bilateral in 23 patients (77%) and in only four patients (13%) was it isolated to the atrophic lobe. The correlation between atrophy and portal vein obstruction was significant, with 90% sensitivity, 97% specificity, and 96% positive predictive value (p < .00001). For the correlation between atrophy and biliary obstruction, the sensitivity of angiography with CT during arterial portography was 90%, specificity was 23%, and positive predictive value was 54% (p = .17). CONCLUSION: Hepatic lobar atrophy usually occurs in the setting of combined biliary and portal vein obstruction. A significant correlation exists between hepatic lobar atrophy and ipsilateral portal vein obstruction.     

Obstructive jaundice due to multiple hepatic abscesses

A 63-year-old Japanese male with diabetes mellitus developed obstructive jaundice following the onset of multiple hepatic abscesses. Percutaneous transhepatic cholangiography showed intrahepatic bile duct irregularity and dilatations accompanied by a complete obstruction of the right branch of the intrahepatic bile duct. Three kinds of organisms were cultured from the blood and the drained bile. The cholangiographic changes returned to the normal after the liver abscesses subsided following biliary drainage and the administration of intravenous antibiotics.     

Composition of biliary lipids and kinetics of bile acids after cholecystectomy in man

Postcholecystectomy biliary lipid composition and bile acid kinetics were studied in 24 women and 4 men. Hepatic bile was collected periodically for as long as 4 months without interrupting the enterohepatic circulation and without infecting the biliary system. In 23 patients with cholesterol gallstones, fasting biliary cholesterol made up 10.2% of total lipids in the steady state; in 5 patients with bilirubinate stones, saturation of fasting hepatic bile with cholesterol was lower (8.7% of total lipids). The percentage of deoxycholic acid after cholecystectomy was not higher than that of seven healthy, noncholecystectomized controls. Postcholecystectomy studies of diurnal variation of biliary lipids (7 patients) showed that postprandial hepatic bile had a significantly lower cholesterol saturation than fasting bile. Pool sizes of cholic and chenodeoxycholic acids were low (average 0.4 g/70 kg, each); total synthesis for both bile acids was normal (average 460 mg/day/70 kg), but fractional turnover rates of the two primary bile acids increased after cholecystectomy, probably due to more frequent recycling of the small bile acid pool.     

Tuberculous pseudotumor causing biliary obstruction. Report of a case with diagnosis by fine needle aspiration biopsy and bile cytology

Hepatobiliary tuberculosis is a rare but distinct clinical entity. We report an unusual case of biliary tract obstruction due to localized hepatic tuberculosis with periportal tuberculous adenitis. The lesion mimicked a malignancy clinically and radiologically. Fine needle aspiration biopsy revealed granulomas, epithelioid histiocytes and Langhans' giant cells. The cytodiagnosis was confirmed by identification of acid-fast bacilli in the bile cytology and isolation of Mycobacterium tuberculosis by culture. The patient responded to antituberculosis therapy. The usefulness of bile cytology in the diagnostic management of biliary tract obstruction is illustrated.     

Expression of the transcriptional regulator Egr-1 in experimental glomerulonephritis: requirement for mesangial cell proliferation

BACKGROUND/AIMS: The hepatic transport of bile salts can be regulated by changes in bile salt pool size and/or in the flux of bile salts through the liver. Prolonged bile salt pool depletion is associated with down-regulation of maximum taurocholate transport and decreased canalicular membrane specific bile salt binding sites. This study was undertaken to investigate: a) whether adaptive down-regulation of maximum hepatic bile salt transport occurs to the same extent for bile acids of different hydrophobicity; and b) the role of microtubule-dependent vesicular pathway in the adaptive changes of bile salt transport capacity. METHODS: Male rats were subjected to 24-h or 48-h external biliary diversion to induce bile salt pool depletion. Basal bile flow, bile salt secretion and lipid secretion, maximum secretory rate of three bile salts of different hydrophobicity (tauroursodeoxycholate, taurocholate and taurochenodeoxycholate) and changes in the biliary excretion of two markers of the microtubule-dependent vesicular pathway (horseradish peroxidase and polyethyleneglycol molecular weight-900) were measured in control and bile salt-depleted rats. Taurocholate-stimulated horseradish peroxidase biliary excretion was also assessed in order to define whether the restoration of bile salt flux across the hepatocytes increased the excretion of this marker in bile salt-depleted rats. RESULTS: The reduction in the maximum secretory rate of the three bile salts under study observed after prolonged biliary diversion was clearly related to their hydrophobicity, with greater reduction for taurochenodeoxycholate and smaller reduction for tauroursodeoxycholate, compared with taurocholate. The biliary excretion of vesicular transport markers was significantly reduced in bile salt-depleted rats. However, when stimulated by taurocholate, biliary excretion of horseradish peroxidase was similar to controls. CONCLUSIONS: The magnitude of the decrease of the hepatic bile salt maximum transport capacity seen after bile salt pool depletion is directly related to the hydrophobicity of the bile salt infused. A functionally depressed vesicular transport pathway appears to be also a contributing factor to this phenomenon.     

Indicators of liver excretory function in patients undergoing biliary decompression for obstructive jaundice

BACKGROUND/AIMS: The recovery of liver function after biliary drainage in patients with obstructive jaundice may be different depending on the severity and duration of the obstruction. We conducted this study to determine whether there are any clinical factors that can be used to monitor the course of recovery. METHODOLOGY: Serum and bile from 12 patients were collected for biochemical testing on the day of drainage and every 3 days for 6 days. Liver function was evaluated by the indocyanine green retention test (ICG R15) before and 6 days after decompression. Patients with an ICG R15 reduction ratio of less than 50% were considered to have a poor recovery (group 1, n = 6), while a good recovery was indicated by a reduction ratio higher than 50% (group 2, n = 6). Sequential data were compared between the groups and correlated with the results of the ICG test. RESULTS: After drainage, the patients in group 1 had less bile acid excretion on day 3 (1.0 +/- 0.8 vs. 3.4 +/- 1.1 mmol/day, p < 0.05), a slower reduction ratio of serum bilirubin on day 3 (0.38 +/- 0.14 vs. 0.60 +/- 0.12, p < 0.05) and more biliary output on day 6 (1.11 +/- 0.25 vs. 0.60 +/- 0.25 L/day, p < 0.05). The ICG R15 reduction ratio was well correlated with the bilirubin reduction ratio, the bile volume and the amount of excreted bile acids checked on day 3 (gamma = 0.73, -0.71 and 0.74, respectively, p < 0.01). CONCLUSIONS: The presence of choleresis implies ductular cell hyperplasia, while decreased excretion of bile acids and a slow reduction of hyperbilirubinemia represents severe liver damage. Both conditions are sequelae of prolonged obstruction; therefore, they might indicate a long and poor recovery.     

Orthotopic transplantation of a partial hepatic autograft in dogs

The purpose of this study was to investigate the availability of an orthotopic transplantation of partial hepatic autograft in dogs as a means of surgical training. Male mongrel dogs weighting 10-15 kg were used. The left lobe of the liver was harvested while preserving the left branches of the portal vein, hepatic artery and bile duct, and the left hepatic vein. The remnant liver was removed while preserving the inferior vena cava using a veno-venous bypass. Orthotopic transplantation of the autograft was performed while anastomosing the left hepatic vein to the inferior vena cava, portal and arterial reconstruction, and external biliary drainage. Thirteen out of 29 dogs survived more than 48 h after transplantation. However, 6 out of 13 dogs were sacrificed after developing bile peritonitis due to a dislodgement of the biliary catheter, and only two dogs were able to survive for 7 days after transplantation. The arterial ketone body ratio recovered to 1.0 within 1 h after reperfusion, and the ratio of the dogs that survived for more than 48 h remained above 1.0 until sacrifice. Orthotopic transplantation of a partial hepatic autograft is a useful and simple procedure to train surgeons for partial liver transplantation.     

Hepatocellular carcinoma presenting as a tumour of the hilar and extrahepatic bile ducts

A case of hepatocellular carcinoma extending within the large extra- and intrahepatic bile ducts is reported. No primary tumour was found in the liver parenchyma by abdominal ultrasound, spiral computed tomography or magnetic resonance, but transduodenal cholangioscopy showed tumour in the common hepatic ducts and the two main branches. Endoscopic biopsy showed highly differentiated hepatocellular carcinoma. The patient was treated with endoscopic biliary drainage and died at home 7 months after admittance.     

Consequences of cholestasis from the pathologist's viewpoint

The present review summarizes extrahepatic obstructive cholestasis and its hepatic morphological sequelae. The first part focuses on early and late changes visualized in liver biopsies. In the second part, pathogenic mechanisms involved in liver fibrosis are discussed. A third section deals with the morphological differentiation of liver atrophy occurring in obstruction and secondary biliary cirrhosis. Biliary obstruction induces a wide array of typical early and late changes of liver morphology. They comprise the visible accumulation of bile in hepatocytes and macrophages, liver cell damage (cholate stasis; apoptosis), alterations of cellular organelles, cytoskeleton and junctions, portal tract edema with cellular infiltrates, ductular reaction, bile infarcts, bile extravasation and, in the later stages, fibrosis and partial nodular change. The pathogenesis of septal fibrosis appears to involve, besides fibroblasts, myofibroblasts probably derived from the Ito cell system. Myofibroblasts seem to closely follow proliferating ductules which, therefore, have a crucial role in hepatic tissue remodeling. There are considerable structural similarities between liver atrophy occurring in biliary and/ or portal vein obstruction and secondary biliary cirrhosis. In both situations partial nodular change ensues, but with preserved vascular relations. In addition, both changes are partially reversible in the early stages of the disease. Irreversible late stages of atrophy and secondary biliary cirrhosis are, therefore, maximum alterations of a similar, dynamic process, involving part of the liver in the first case and the whole organ in the second, depending on the level of obstruction.     

Effect of P-450 inducers on biliary excretion of glutathione and its hydrolysis products. Correlation between hepatic gamma-glutamyltranspeptidase activity and the proportion of glutathione hydrolysis products in bile

This study was designed to determine if a relationship exists between hepatic gamma-glutamyltranspeptidase (gamma-GT) activity and the biliary excretion of glutathione (GSH) and its hydrolysis products. Rats were pretreated with the following microsomal enzyme inducers: pregnenolone-16 alpha-carbonitrile (PCN), dexamethasone (DEX), 3-methylcholanthrene, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), phenobarbital (PB), ethanol (ETOH), trans-stilbene oxide (TSO), butylated hydroxyanisole (BHA), isosafrole (ISF), clofibrate, and benzo(a)pyrene. Hepatic gamma-GT activity was quantitated spectrophotometrically; bile and liver samples were analyzed by HPLC for reduced and oxidized GSH and their hydrolysis products (cysteine, cysteinylglycine, and cysteinylglycine disulfide). Administration of the inducers had only minor effects on hepatic GSH concentration, as BHA was the only agent to increase GSH concentration. However, these inducers had a pronounced effect on the biliary excretion of total thiol-derived sulfur as PCN, PB, and ISF produced an increase, whereas TCDD, ETOH, and TSO caused a decrease. The relative amount of the GSH hydrolysis products in bile was highly dependent on gamma-GT activity. For example, hepatic gamma-GT activity was increased by PCN, DEX, BHA, TSO, and ISF. They also increased the GSH hydrolysis products to total thiol-derived sulfur ratio in bile. In conclusion, the ratio of GSH hydrolysis products to total thiol-derived sulfur excreted in rat bile reflects the hepatic gamma-GT activity.     

Biliary reabsorption of cholate sodium, glycocholate sodium, and taurocholate sodium from the rat biliary tree after retrograde intrabiliary injection

The results demonstrate the biliary reabsorption of 14C-cholate, 14C-glycocholate, and 14C-taurocholate from the rat biliary tree after retrograde intrabiliary injection. It could be shown that retrograde injection of these bile salts (20 nmol) in a total volume of 40 mul leads to significantly increased biliary reabsorption in contrast to the administration in a retrograde volume of only 20 mul. These differences in reabsorption may be explained by greater reabsorption at a more proximal site in the biliary tree. Furthermore 14C-glycocholate and 14C-taurocholate are reabsorbed to a lesser extent in contrast to 14C-cholate when bile flow was restarted at once after retrograde injection in a volume of 40 mul. It is speculated that conjugation of cholate to glycine and taurine has some effect on the extent of biliary reabsorption of this bile acid. Following the results presented in this paper one might hypothize that biliary reabsorption has an important influence on bile composition i.e. the biliary excretion of bile salts.     

Time-related changes of cold-stored rat liver in University of Wisconsin solution. Effect of ursodeoxycholate

Livers of Wistar rats were stored between 0 and 36 hrs. in the University of Wisconsin preservation liquid in order to determine time-related biochemical and morphological hepatic changes. Ursodeoxycholate (100 microM) was also added in the medium to test the hepatoprotective properties of the bile salt. Biochemical assays were performed on hepatic microsomes, plasma and biliary canalicular membranes. Protein and lipid composition of the microsomal and baso-lateral plasma membranes remained stable. Protein and cholesterol content of the biliary canalicular membranes decreased, phospholipid/cholesterol ratio increased between 0 and 36 hrs.; it resulted in a leak of 5'-nucleotidase and leucine amino peptidase activity of these biliary canalicular membranes, especially up to 12 hrs. Between 0 and 36 hrs., the lipid and protein content remained stable in the plasma membranes, as well as both tested enzymatic activities. Observations under electron microscopy showed alterations and underlined fragility of the bile canaliculi, particularly after 24 hrs. preservation. Ultrastructure of sinusoidal membranes showed damaged microvilli. Endoplasmic reticulum remained unchanged, in relation to the stability of the microsomal lipidic, proteic content and hydroxymethylglutaryl-coenzyme A reductase activity, except the decreased protein content after preservation for 36 hrs without ursodeoxycholate. Ursodeoxycholate by itself did not protect against the described disturbances.     

Ultrastructure of the glial cells after spinal cord compression in rabbits

Sump syndrome is a rare complication of biliary-enteric anastomosis. Classically, the distal bile duct becomes obstructed by gastrointestinal debris after choledochoduodenostomy, resulting in cholangitis or, less commonly pancreatitis. Obstruction of the biliary tree by gastrointestinal contents after Roux-en-Y choledochojejunostomy or hepaticojejunostomy has not been described in the English-language literature. This report details the diagnostic and operative management of the first patient with sump syndrome after hepaticojejunostomy. The presumed pathophysiology was reflux of vegetable matter up the efferent limb, resulting in hepatic duct obstruction and cholangitis. The patient ultimately required complex choledochoscopic drainage of the intrahepatic biliary tree and revision of the previous Roux-en-Y hepaticojejunostomy.     

Nutritional value and antinutritional factor content of precooked flours prepared from Canavalia ensiformis

There is a great body of evidence linking a high fat diet with the formation of gallstones. However, the effect of fat per se on obstructive liver damage (not involving gallstone formation) has not been assessed. The aim of this work was to study the effect of a high fat diet on liver damage induced by bile duct ligation in rats. Male 21-day-old Wistar rats were divided into two groups: group 1 received standard Purina chow diet 5001 containing 4.5% fat, group 2 received Purina chow diet 5001 enriched with 33% pork fat. Animals were allowed food and water ad libitum for 5 weeks. Obstructive jaundice was induced by double ligation and division of the common bile duct. The animals were sacrificed 1 week after biliary obstruction. Control animals were sham operated. Serum bilirubins and alkaline phosphatase, gamma-glutamyl transpeptidase and glutamic pyruvic transminase enzyme activities increased by biliary obstruction. Glycogen content decreased in the bile duct-ligated rats. These effects were more important in the group fed a 33% fat diet. Our results show that a high animal fat diet increases liver damage in experimental biliary obstruction in rats. Owing to our experimental design (bile duct ligation), the effect of a high fat diet cannot be attributed to an increase in the formation of gallstones but a direct effect must be considered. The mechanism by which fat augmented liver damage can be associated with an increase of total bile content and its toxicity.     

Cholestasis secondary to Hodgkin's disease: report of 2 cases of vanishing bile duct syndrome

Only a small percentage of patients with Hodgkin's disease become clinically Jaundiced during their disease. This Jaundice may be secondary to biliary obstruction, hemolysis, direct hepatic infiltration by the disease, drug toxicity or viral hepatitis. Vanishing bile duct syndrome secondary to Hodgkin's disease is a rare cause of cholestasis in these patients, only 13 cases having been reported so far. The authors describe 2 patients who developed severe Jaundice secondary to Hodgkin's disease due to vanishing bile duct syndrome affecting small intrahepatic bile ducts.     

Clinical factors associated with positive bile cultures during primary percutaneous biliary drainage

PURPOSE: To evaluate the utility of routine bile cultures and to determine the risk factors for bacterial colonization of the bile as well as the biliary flora in patients with biliary obstruction undergoing primary percutaneous biliary drainage. MATERIALS AND METHODS: Between October 1995 and January 1997, bile cultures were prospectively obtained in all patients undergoing percutaneous biliary drainage. Seventy-six patients underwent 86 procedures. Culture results were correlated with clinical, laboratory, and demographic variables. The antibiotic sensitivities of cultured organisms were examined. RESULTS: Fever, previous endoscopic or percutaneous biliary instrumentation, and bilioenteric anastomosis were significant predictors of a positive bile culture. In the absence of any of these indicators, bile cultures were unlikely to be positive. Enterococcus species was the organism isolated most commonly. Yeast, gram-negative aerobic bacilli, and Streptococcus viridans followed in frequency. CONCLUSION: Bile cultures provide valuable information that was useful for planning antibiotic prophylaxis and treatment. The likelihood of positive bile cultures can be predicted based on certain clinical variables. Continued investigation is needed to better predict bacterial flora in individual patients. Given the association between previous instrumentation and biliary colonization, noninvasive imaging modalities should be exhausted before invasive procedures are performed for solely diagnostic purposes in patients with biliary obstruction.