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PubMed1982

A simple reverse-phase high pressure liquid chromatographic determination of conjugated bile acids in serum and bile using a novel radial compression separation system.

A new HPLC method for analyzing conjugated bile acids in bile and especially in serum is presented. For separation of novel radial compression system with a 10-cm flexible-walled reverse phase column (RCM-100 Module, Waters Ass.) packed with micro-Bondapak C18 was used. Without any laborious hydrolysis and derivatization steps, 10 conjugated bile acids are separated easily with a mobile phase of methanol/0.01 mol/1 KH2PO4 (150:50, v/v; pH 6.0) in about 30 min, determined with a variable UV detector at 200 nm and a flow rate of only 0.5 ml/min. Dexamethasone was used as internal standard. Conjugated bile acids are extracted from serum with Sep-pak C18 cartridges after a 1:8 dilution with a mixture of the mobile phase and 0.2 mol/l NaOH (6:8, v/v). Extraction from bile was also performed using Sep-pak cartridges after a 1:20 dilution with 0.5 mol/l phosphate buffer (pH 7.0). The complete analysis time of the method, with Sep-pak extraction, is less than one hour. Recoveries for the different conjugated bile acid varied from 89.4% to 96.7% in serum. The HPLC method for serum and bile was validated for the primary bile acids cholate and chenodeoxycholate against the previously described GLC method. Agreement between the two methods was excellent. Duplicate analyses in serum and bile samples also showed that the reproducibility was good.

Bile↗

PubMed1989

Bile salts and hydrodynamics of bile formation.

We report a novel method to assess bile secretory pressure using a Statham pressure transducer. The studies were performed in vivo in male Sprague-Dawley rats under pentobarbital anesthesia. Maximal secretory pressure averaged 21.8 +/- 1.1 (S.D.) cmH2O. The bile accumulated after 10 min of obstruction was 7.7 +/- 2.8 microliters.g-1; assuming a basal biliary dead space of 2.3 microliters.g-1, the distended capacity of the biliary tree averaged 10 microliters.g-1. The small volume of the strain gauge permitted calculation of compliance of the biliary tree which averaged 0.35 +/- 0.12 microliters.cmH2O-1.g-1. Stimulation of bile flow by bile salts reduced the time required to reach maximal bile secretory pressure. Taurocholate but not taurodehydrocholate decreased maximal secretory pressure within minutes, the pressure-time curves showing a new equilibrium between bile formation and regurgitation forces. Both bile flow and bile salt recoveries were decreased by taurocholate but not by taurodehydrocholate. Taurocholate decreased biliary compliance while taurodehydrocholate had no effect. This provides further evidence that taurocholate increases the biliary permeability and suggests that this bile salt also affects the elastic properties of the biliary tree.

Animals↗

PubMed2004

Characterization of bile acids and fatty acids from ox bile in oil paintings by gas chromatography-mass spectrometry.

Characterization of ox bile, traditionally used in painting, is of interest in the fields of archaeometry and conservation and restoration of works of art. Bile acids, fatty acids (F), and cholesterol found in ox bile have been identified using a derivatization method that combines the formation of ethyl esters from the carboxylic groups and the trimethylsilyl ethers from hydroxyl groups. This method of analysis is consistent with these others proposed by the authors to analyze drying oils, proteins, and diterpenic resins usually used as binders and varnishes by the painters. Bile acids from binary samples such as animal glue/ox bile, casein/ox bile and Arabic gum/ox bile have been successfully analyzed using the proposed method. Finally, a method of analysis of mixtures of drying oil and ox bile has been also proposed attempting to quantitatively characterize samples in which ox bile was added to the drying oil for increasing the surfactant properties.

Animals↗

PubMed1996

Bile-pancreatic juice replacement not cholinergic- and cholecystokinin-receptor blockade reverses acinar cell hyperstimulation after bile-pancreatic duct ligation.

BACKGROUND: Acinar cell inhibitors (eg, atropine) fail to ameliorate clinical and experimental acute pancreatitis. We hypothesized that amelioration of pancreatic acinar cell hyperstimulation after bile and pancreatic duct ligation is better with gut replacement of bile and pancreatic juice than with cholinergic- and cholecystokinin (CCK)-receptor blockade. METHODS: Using acinar cell amylase activity as an index of hyperstimulation, we studied 63 rats in two sets of experiments. Bile-pancreatic juice exclusion from gut--without (set one) and with (set two) bile and pancreatic duct obstruction--was treated with atropine and CCK-receptor antagonist L-364,718, or with enteral replacement of bile-pancreatic juice. RESULTS: In the set one experiment, acinar cell hyperstimulation after bile-pancreatic juice exclusion was reversed by combined L-364,718 and atropine pretreatment. In set two, acinar cell hyperstimulation after bile and pancreatic duct ligation was reversed by enteral bile and pancreatic juice replacement, but not by combined L-364,718 and atropine pretreatment. CONCLUSIONS: According to this experimental corollary of early gallstone impaction, prevention of acinar cell hyperstimulation after duct occlusion should be aimed at the source of the response to bile-pancreatic juice exclusion, namely, the gut, rather than at the target of the response, the pancreatic acinar cell.

Amylases↗

PubMed1986

Naturally occurring conjugated bile acids, measured by high-performance liquid chromatography, in human, dog, and rabbit bile.

The aim of this study was to determine the biliary pattern of conjugated bile acids after stimulation of their enterohepatic circulation. Conjugated bile acids were separated by reversed-phase ion-pair chromatography without prior derivatization. A MicroPak SP-C18-IP-4 column was used as non-polar matrix, and an ionic alkyl compound, tetrabutylammonium phosphate, was added to the mobile phase, which was a mixture of acetonitrile and water. Quantification was made by UV absorption at 210 nm with external standardization. In fourteen human patients with external biliary drainage after papillotomy there was preferential glycine conjugation. The mean values were 36.5% for glycocholic acid, 33% for glycochenodeoxycholic acid, and 10.0% for glycodeoxycholic acid. Only 15.2% of the biliary bile acids were taurine metabolites. Conjugates of ursodeoxycholic acid were below 2.1%. In most cases, conjugated lithocholic acid was not detected. Within 4 h after ingestion of a standardized meal there were no significant changes in the biliary bile acid pattern. In four dogs (beagles), glycine-conjugated bile acids were lacking. The mean values were 74.3% for taurocholic acid, 14.9% for taurodeoxycholic acid, and 5.3% for taurochenodeoxycholic acid. In six rabbits, 87.4% of biliary bile acids was identified as glycodeoxycholic acid and 5.3% as glycocholic acid. In conscious dogs, as well as in rabbits, the stimulation of biliary secretion by cholecystokinin and/or secretin had no effect on the biliary bile acid spectrum. Evidently, there is a difference in the biliary composition of conjugated bile acids between humans, dogs, and rabbits. Because of the different physicochemical behaviour of glycine- and taurine-conjugated bile salts, it seems difficult to compare the therapeutic effect of gallstone dissolution in various species.

Animals↗

PubMed1999

Role of bile salt hydrophobicity in distribution of phospholipid species to carriers in supersaturated model bile solutions.

BACKGROUND: Phospholipid species modulate cholesterol-holding capacity and, therefore, regulate bile metastability. METHODS: In this study, we investigated the effect of bile salt hydrophobicity on the distribution of phospholipids among lipid particles in supersaturated model bile solutions (total lipid concentration, 9 g/dL; taurocholate/phospholipid ratio 3.0, cholesterol saturation index 1.3), by using gel permeation chromatography. RESULTS: With an increase of bile salt hydrophobicity in the elution buffer, the uptake of cholesterol and phospholipids into bile salt micelles was increased, associated with an increased cholesterol/phospholipid molar ratio of the vesicles. In contrast, there was an inverse correlation between the hydrophobicity of the phospholipid species in the vesicles and that of bile salts in the elution buffer, suggesting that hydrophobic bile salts induced preferential uptake of hydrophobic phospholipids into bile salt micelles, while less hydrophobic phospholipids, with a relatively low cholesterol-holding capacity, remained in the vesicles. CONCLUSIONS: These data indicate that bile salt hydrophobicity regulates vesicular cholesterol metastability by modulating the hydrophobicity of phospholipids in vesicles, as well as the lipid distribution among various biliary lipid particles.

Bile↗

PubMed1994

Dietary psyllium hydrocolloid and pectin increase bile acid pool size and change bile acid composition in rats.

Bile acid composition, synthetic rate and pool size were determined in rats fed diets containing 5 g cellulose, 5 g pectin or 5 g psyllium hydrocolloid/100 g diet using the washout technique. Bile acid pool sizes were similar for pectin- and psyllium-fed rats, and both were higher than the pool size for rats fed cellulose (5.48 +/- 1.22, 4.70 +/- 0.71 and 1.77 +/- 0.41 mumol/100 g body wt, respectively). Bile acid secretion rates showed a similar pattern [1730 +/- 496, 1551 +/- 252 and 572 +/- 88 nmol/(h.100 g body wt)], as did basal synthetic rates [129 +/- 25, 126 +/- 42 and 87 +/- 18 pmol/(h.100 g body wt)]. Individual and total bile acid pool sizes were generally higher in animals fed the pectin- or psyllium-supplemented diet compared with rats fed cellulose. Pectin or psyllium consumption resulted in a lower hydrophobicity of the bile acid pool and a lower ratio of circulating 12 alpha-hydroxylated to non-12 alpha-hydroxylated bile acids compared with cellulose consumption. This reduced hydrophobicity has been shown to reduce feedback inhibition of bile acid synthesis and may be responsible for the larger bile acid pool size. Changes in the location of reabsorption of bile acids may also be responsible for changes in the pool size and composition. These changes are characteristic of greater sterol excretion.

Animals↗

PubMed1994

Effects of bile salt and phospholipid hydrophobicity on lithogenicity of human gallbladder bile.

Increased biliary bile salt and phospholipid hydrophobicity may promote nucleation of cholesterol crystals and gallstone formation. We therefore compared bile salt composition (determined by gas-liquid chromatography) in patients with cholesterol (n = 35) and pigment (n = 16) gallstones (group A). Bile salt composition and cumulative bile salt hydrophobicity index were not different between both stone types. Hydrophobicity index or % of individual bile salts did not correlate with cholesterol saturation index or nucleation time. In an additional 21 cholesterol stone patients (group B) biliary bile salt and phospholipid hydrophobicity as determined by high-pressure liquid chromatography did not correlate with cholesterol saturation index or nucleation time. In both group A and group B, cholesterol stone patients with cholesterol crystals in their fresh biles had a higher % deoxycholic acid, a lower % cholic acid and a higher bile salt hydrophobicity index than crystal-negative patients. This study indicates the need for further research on the role of bile salt hydrophobicity in the pathogenesis of gallstones.

Adult↗

PubMedDate not supplied

Regurgitation of bile acids in rat liver under bile drainage: quantitative analysis by taurine or ursodeoxycholate loading test.

In rats with an interrupted enterohepatic circulation of bile acids, levels of serum taurine-conjugated bile acids were increased significantly 3 h after intravenous administration of taurine. Similarly, serum taurine- or glycine-conjugated ursodeoxycholic acid (UDCA) was increased significantly 2 h after UDCA administration. These findings suggested that the administered taurine or UDCA was taken up into hepatocytes and utilized to form conjugated bile acids, which were thereafter regurgitated into the systemic circulation from the liver. The proportion of regurgitated taurine-conjugated bile acids relative to total serum bile acids measured by taurine loading (30%) almost coincided with that of regurgitated taurine- or glycine-conjugated UDCA relative to total serum bile acids measured by UDCA loading (31.6%). Thus, the present study showed conclusively that at least 30% of serum bile acids are derived from newly conjugated bile acids that are regurgitated from the liver in rats with bile fistula.

Animals↗

PubMed1993

Reduced cholesterol metastability of hepatic bile and its further decline in gall bladder bile in patients with cholesterol gall stones.

The reduced metastability of biliary cholesterol in the gall bladder bile of patients with cholesterol gall stones has been well shown. The purpose of this study was to examine the hypothesis that such a difference in metastability already exists in hepatic bile. Paired hepatic and gall bladder bile samples were collected from 10 patients with cholesterol gall stones and six patients without gall stones. Cholesterol nucleation time, biliary lipid concentration, vesicular cholesterol distribution, and biliary protein concentration were measured and compared. The nucleation time in the hepatic bile of patients with cholesterol gall stones was significantly shorter than the gall stone free patients (8.2 (7.2) v 15.7 (5.8) days, p < 0.05), and was associated with a greater concentration of biliary lipid despite the lack of a difference in the cholesterol saturation index (CSI) and total protein concentration. During the storage of bile in the gall bladder, the nucleation time became quicker in the patients with cholesterol gall stone (2.9 (1.7) days) while it was similar in the gall stone free patients (17.3 (5.7) days) compared with that of the corresponding hepatic bile. These differences were associated with a higher CSI (1.44 (0.33) v 1.13 (0.14), p < 0.05) and a greater vesicular cholesterol distribution (19.7 (11.9) v 4.4 (1.4)%, p < 0.01) in the patients with cholesterol gall stones than the gall stone free patients. The concentrations of total lipid and protein in gall bladder bile were not significantly different between the two groups. In conclusion, patients with cholesterol gall stones produce less metastable hepatic bile by the evidence of shorter nucleation time. During the storage of the bile in the gall bladder, the metastability is reduced further only in the cholesterol gall stone patients but not in the gall stone free patients.

Bile↗

PubMed1979

Bile flow and electrolyte composition of bile associated with maximum bilirubin excretion in sheep.

Changes in the composition of bile accompanying the maximum biliary excretion (Emax) of bilirubin were investigated in sheep. Sheep fitted with chronic 'T-tubes' in the common bile duct were infused with taurocholate and bilirubin at various rates. Bile collected during both pre- and post-bilirubin steady-state periods was analyzed for the biliary concentration of electrolytes, bile salts, and bilirubin. Bilirubin Emax was 24.6 mumol/min while bile salt excretion during this period was 103 mumol/min. At Emax bilirubin entry into bile reached a concentration of 16.1 mumol/mL, increased the biliary concentration of sodium, did not change osmolarity of bile, and did not increase bile flow. The data suggest that bilirubin either interacts with mixed micelles in bile or forms molecular aggregates.

Animals↗

PubMed1983

Effect of bile and bile acids on binding of intrinsic factor to cobalamin and intrinsic factor-cobalamin complex to ileal receptor.

Human bile or commercially available bile acids bind [57Co]cobalamin (Cbl) due to the presence of "R"-type binders, which are normally present in the former and present as a contaminant in the latter. The competition of these R proteins for the binding of Cbl by intrinsic factor (IF) could explain the in vivo inhibition attributed previously to the bile acids themselves. R protein seems to be involved in the inhibition of Cbl binding because protease digestion of either bile or bile acid abolishes the Cbl binding ability. Moreover, antibody to R protein abolishes the inhibition. Bile or bile acids do not have a direct effect on either purified IF or the IF-Cbl receptor molecule, even though bile acids increase the attachment of IF-[57Co]Cbl to ileal brush-border membranes. These data demonstrate two steps where components of bile could affect Cbl absorption: the binding of Cbl to IF and of IF-Cbl to its ileal receptor. It is not clear whether these in vitro phenomena are important for the normal absorption of Cbl in vivo.

Animals↗

PubMedDate not supplied

Bile acid load on the DNA distribution pattern of bile ductules and cholangiocarcinoma induced by diisopropanolnitrosamine in hamsters.

This study evaluated the influence of bile acid load on the DNA distribution pattern of proliferated bile ductules and cholangiocarcinoma induced by diisopropanolnitrosamine. Ninety hamsters were separated into control, tauro- and deoxycholic acid (DCA) groups. The DNA distribution pattern of intrahepatic lesions at 15-25 weeks was measured by cytofluorometry and classified into three types: I (-A, -B), II and III, according to the degree of dispersion on the DNA histogram. Regarding proliferated bile ductule lesions, all groups showed an increase in cell populations, indicating the dispersion of nuclear DNA content from the 4C to 6C ranges over the course of 25 weeks, and two groups with bile acids, especially the DCA group, revealed significant high incidences of lesions with type I-B plus II compared with those in the control group (p < 0.05, 0.01). Changes in carcinoma types were similar to those of bile ductule lesions, and the tumors in the DCA group had a significant high frequency of type II plus III (p < 0.05). In addition, heterogeneity of the DNA distribution pattern was observed within individual lesions of not only carcinoma but also bile ductules. These results suggest that bile acid load, especially DCA, promotes an increase in nuclear DNA content or DNA polyploidization and enhances the distribution of the DNA pattern of proliferating bile ductules and carcinoma. Furthermore, a bile ductule-carcinoma sequence may be present in the development of cholangiocarcinoma.

Animals↗

PubMed2002

Helicobacter species are not detectable by 16S rDNA PCR in bile from Dutch patients with common bile duct stones.

BACKGROUND: Some Helicobacter species colonize the intestinal tract. To explore the possible relation between Helicobacter spp. and gallbladder disorders, we have investigated their presence in bile of patients with biliary obstruction and dilatation of the bile ducts. METHODS: Bile was sampled from 31 Dutch patients with biliary obstruction identified by jaundice and dilatation of the bile ducts on ultrasound. Samples (n = 31) were obtained immediately following cannulation of the common bile duct (CBD) by endoscopic retrograde cholangiopancreatography (ERCP) (n = 29) or by peri-operative puncture of the gallbladder (n = 2). DNA was isolated from bile by binding to diatoms. Helicobacter spp. were detected by a sensitive (detection limit 1 CFU per reaction tube) 16S rDNA PCR with genus-specific primers. Duplicate samples were spiked with Helicobacter pylori DNA and subjected to PCR in order to check for inhibition. RESULTS: 28 patients had CBD stones (bile collected by ERCP (n = 26) or operatively (n = 2)), 2 had a pancreatic head tumor, and in 1 no abnormalities were found. In 1 of 21 amplifiable bile samples (10/31 inhibited) from Dutch patients with CBD stones, H. pylori 16S rDNA was found. CONCLUSION: Our data indicate that CBD stones in Dutch patients are not associated with the presence of Helicobacter spp. in bile.

Adult↗

PubMed1989

Role of primary and secondary bile acids as feedback inhibitors of bile acid synthesis in the rat in vivo.

The effect of various primary and secondary bile acids on the rates of synthesis of all major bile acids was studied in the live rat with an extracorporal bile duct. Bile acid synthesis was determined using HPLC based on mass or by isotope dilution. Derepressed rates of bile acid synthesis (30-54 h) were inhibited by an infusion of taurocholic acid only at a supraphysiological dose of 500 mumol/kg per h, but not at 300 mumol/kg per h, which approximates the initial bile acid secretion (250 mumol/kg per h). When administered together with taurocholic acid (200 mumol/kg per h) only a high dose of taurochenodeoxycholic acid (100 mumol/kg per h) decreased taurocholic but not tauromuricholic or taurochenodeoxycholic acid synthesis. The only bile acid suppressing taurocholic acid (36-71%) and taurochenodeoxycholic acid (up to 33%) formation at an infusion rate close to the normal portal flux was deoxy- or taurodeoxycholic acid at 15-50 mumol/kg per h. It may be concluded that deoxycholic acid and possibly other secondary bile acids are much more potent inhibitors than primary bile acids.

Alanine Transaminase↗

PubMed1999

[The detergent properties of the bile acids and their action on bile secretion].

We investigated the influence of the hydrophilic bile acids (cholic and ursodeoxycholic) and hydrophobic (chenodeoxycholic and deoxycholic) on the bile secretion during the isolated rat liver perfusion to find out the role of the bile acids detergent properties in their choleretic and cholestatic effects. The increase of the concentration of the hydrophobic bile acids changed the type of the bile secretion rate dynamic from one- to two-peak stimulation. It was shown that the difference in the rate of the bile acids biotransformation was essential both in this process and in the possibility of the bile acids to cause the cholestasis. It was evaluated the bile acids osmotic activity under the micelle formation and micelle information conditions. It was shown the significance of the other bile acids effects but direct osmotic on the osmotic gradient development.

Animals↗

PubMedDate not supplied

Effect of 4-hydroxyandrost-4-ene-3,17-dione (formestane) on the bile secretion and metabolism of 4-(14)C-cholesterol to bile acids.

4-Hydroxyandrost-4-ene-3,17-dione (formestane) is a selective aromatase inhibitor. It is indicated for postmenopausal patients with advanced breast cancer. The aim of the present study was to investigate the effect of 4-hydroxyandrost-4-ene-3,17-dione on the bile secretion and metabolism of 4-(14)C-cholesterol to bile acid. The experiments were carried out in the ovariectomized and sham-operated female Wistar rats. Formestane (20 mg/kg, i.m., daily) was administered to animals for 2 weeks. Twenty four hours after the last drug administration, rats were anesthetized with ethyl urethane. 4-(14)C-cholesterol (740 kBq/kg, s.a. 2.28 GBq/mmol) was infused for 1 min by catheter inserted into the jugular vein. Bile samples were assayed for total 14C radioactivity 14C-bile acids were determined in bile (after thin-layer chromatographic separation) by the use of isotopic technique with liquid scintillator. Previous studies showed that systemic adverse effects occurred in about 12% of patients following intramuscular drug administration. Many of them such as hot flushes, vaginal spotting and emotional lability were related to the mechanism of action of formestane i.e. estrogen suppression. Lethargy, rash, nausea, dizziness, indigestion, ataxia, cramps and facial swelling have also been reported. The results of the present study have shown that formestane administered to the female ovariectomized rats decreased the bile secretion and diminished conversion of 4-(14)C-cholesterol to trihydroxy bile acids. The decreased synthesis of trihydroxy bile acids and increased concentrations of cholesterol and litocholic acid in bile may be associated with increased risk of gallstone formation.

Androstenedione↗

PubMed1989

Bacteriological study of choledochal bile in patients with common bile duct stones, with or without acute suppurative cholangitis.

Acute suppurative cholangitis is a serious complication in extrahepatic biliary tract obstruction. Bacteriological studies have shown that in these patients the bile cultures usually present several kinds of bacteria, especially Gram-negative aerobic bacteria such as E. coli and Klebsiella sp. The mortality rate in our patients with common bile duct stones and acute suppurative cholangitis is enhanced 5- to 10-fold. The aim of the present study was to analyse the bacterial properties of choledochal bile in a large number of patients with common bile duct stones, with or without acute suppurative cholangitis, and to determine the sensitivity of the isolated bacteria to several antibiotics used in clinical practice. The greatest sensitivity of the bacteria isolated from the common bile duct was seen for aminoglycosides (gentamicin and amikacin) and for cephalosporins, especially of the third generation (cefotaxime, ceftazidime, ceftriaxone and cefoperazone). Ampicillin showed a low sensitivity rate. The appropriate selection of the antibiotics used in acute suppurative cholangitis depends not only on the concentration of the antibiotic in the bile; a high plasma concentration is even more important to control the septic manifestations. A high excretion to the bile can affect the intestinal flora, leading to diarrhea (e. g. with cefoperazone). Ampicillin has been giving contradictory results in respect of bile and plasma concentrations. Gentamicin attains 30-40% of the plasma concentration in the bile, but persists for as long as 8 hours with excellent sensitivity and clinical efficacy. Amikacin reaches 44% of the plasma concentration with a very high sensitivity (96%).(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗