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Characterization of cholylglycine hydrolase from a bile-adapted strain of Xanthomonas maltophilia and its application for quantitative hydrolysis of conjugated bile salts.

Purified bile salt hydrolase from bile-adapted Xanthomonas maltophilia displays Michaelis-Menten kinetics on cholylglycine and cholyltaurine and hydrolyzes bile salts also in crude bovine bile. The protein is a dimer and is resistant to proteinases and to heating at 55 to 60 degrees C for up to 60 min, in agreement with calorimetric data.

Amidohydrolases↗

Selection of patients for bile diversion surgery: use of bile acid measurement in fasting gastric aspirates.

Eighteen patients with dyspepsia and vomiting which followed surgery for peptic ulcer have completed a study to examine the role of diverting bile from the stomach by a Roux-en-Y procedure. Bile regurgitation and mild epigastric pain relieved by vomiting were abolished. Measurements of bile acids in the fasting gastric aspirate were useful in predicting the outcome of surgery; good results were obtained when initially there was reflex into the stomach of more than 120 mumol/hour of bile acids. A wider group of patients than those selected in previous series may benefit from this operation, as good results can be obtained in patients with dyspepsia relieved by alkali and without achlorhydria or gastritis. Endoscopy was repeated one year after Roux-en-Y operation. Erythema of the mucosa was improved, but gastritis did not improve.

Adult↗

Bile acid transformation by the intestinal flora and cholesterol saturation in bile. Effects of Streptococcus faecium administration.

In 8 normal subjects the daily administration for 30 days of at least 750 X 10(6) Streptococcus faecium, a bacteria producing substances acting against Clostridia and other intestinal microorganisms reduced the cholesterol saturation and the molar percentage of deoxycholic acid (DCA) in bile. The percentage of cholic acid (CA) and chenodeoxycholic acid (DCA) increased significantly in feces, whereas lithocholic acid (LCA) decreased from 49.2 to 40.5%. In vitro fecal samples of subjects taking S. faecium transformed more slowly CA to DCA and CDCA to LCA. Moreover, 7-ketolithocholic seems an intermediate step in the biotransformation of CDCA to LCA. During SF administration, the anaerobic bacteria count of feces decreases, whereas aerobic bacteria increase; the changes of fecal flora may reduce the degradation of primary bile acids influencing the bile acid composition in both the bile and the intestine.

Adult↗

Effect of endoscopic sphincterotomy on bile acid pool size and bile lipid composition in man.

The effect of endoscopic sphincterotomy on bile acid pool size and lipid composition was studied in 3 patients with an intact gallbladder and in 7 patients who had previous cholecystectomy. Measurements were made at two time intervals after endoscopic sphincterotomy, early (3-9 days) and late (6-9 months). Patients with an intact gallbladder showed a marked reduction in their total bile acid pool during follow-up examinations (95.3 +/- SD 14.0 vs. 18.6 +/- 8.1 mumol/kg), whereas in the cholecystectomized patients the pool size showed no significant change (29.4 +/- 13.4 vs. 26.6 +/- 11.4 mumol/kg). The reduction in bile acid pool size caused by sphincterotomy in patients with an intact gallbladder did not increase the degree of cholesterol saturation in hepatic bile.

Aged↗

Alterations of rat intestinal disaccharide absorption by bile or conjugated bile acids.

Taurocholate showed different effects on the jejunal sucrose absorption, depending on the concentration of the bile acid: 3 mmol/l taurocholate inhibited to a small extent (-7%), 10 mmol/l taurocholate had no effect and 30 mmol/l taurocholate increased the sucrose absorption by 17%. The sucrose hydrolysis was augmented by 20% in the presence of 30 mmol/l taurocholate. 2 mmol/l taurochenodeoxycholate caused an increase of sucrose hydrolysis(+ 37%) as well as of sucrose absorption (+ 12%) in the jejunum. Pooled rat bile induced a 21% inhibition of the sucrose hydrolysis, while the sucrose absorption was unchanged. The jejunal maltose hydrolysis was unaffected by 3 mmol/l taurocholate, whereas 10 or 30 mmol/l caused a minimal increase (+ 3%). The absorption of the resulting glucose was reduced to 84--92%. In the ileum 10 mmol/l taurocholate did not change the sucrose hydrolysis and absorption, whereas 2 mmol/l taurochenodeoxycholate induced a 30% decrease. The investigated bile acids had no effect on the microscopic structure of the small intestinal mucosa. Hypertonic solutions caused an accumulation and exfoliation of PAS-positive material at the top of the villi, independent of the presence or absence of bile acids.

Animals↗

Lipid transport into bile and role in bile formation.

Biliary lipid secretion is driven by bile salts, the primary metabolites of cholesterol. Transport of bile salts as well as phospholipids and cholesterol is mediated by ATP Binding Cassette (ABC) transporters. Expression of these transporters is regulated in a coordinate fashion by a set of nuclear hormone receptors explaining the old observation of coupling between bile salt secretion and biliary lipid secretion. Although it is now clear which proteins are involved, the molecular mechanism of biliary lipid secretion is still unresolved. In addition, scarce information is available about the systems responsible for intracellular transport of cholesterol, phospholipid and bile salt. These issues form the subject of the review.

ATP-Binding Cassette Transporters↗

An analysis of data on human hepatic bile. Relationship between main bile components, serum cholesterol and serum triglycerides.

Hepatic bile samples were taken from the common duct during interval operations for gallstone disease, performed under standardized conditions. Prior to operation serum cholesterol and triglycerides levels were determined. The concentrations of Cholesterol (Chol), phospholipids (Lip P) and of the three major bile acids (BA) were determined in ninety-seven samples. The data were subjected to statistical analysis. A highly significant rank correlation was found between the Chol- and the Lip P molar fractions. The rank correlation between the Chol molar fraction and the BA/Lip P ratio was highly significantly negative. Reasons are given why the observed intraindividual differences are interpreted as reflecting interindividual changes. The conclusion is that just as in the animal model, a rise of Chol in human hepatic bile is accompanied by a decrease of the BA/Lip P ratio. A significant correlation was found between chenodeoxycholic acid (CDCA), and the ratio between the two other main bile acids (CA/DCA). High lithogenicity was associated with low CDCA- and high DCA values. In a mathematical representation valid in the sense of a rank correlation, the bile/serum Chol ratio rose with increasing DCA values combined with increasing absolute differences between the other acids. Serum triglycerides were negatively correlated with the BA molar fraction and with the absolute concentration of BA. As a result there was an association between serum triglycerides and lithogenicity.

Bile↗

The escape of bile from the intrhepatic biliary tree in acute bile stasis.

The escape of fluorescent dyes from the bile passages during bile stasis and after a retrograde infusion into the common bile duct was examined in rats and mice. After an intravenous dye injection in bile stasis a strong fluorescence is seen in the periportal spaces. During retrograde dye infusion great amounts of dye accumulate in the periportal spaces but, in consequence of focal disruption of liver cell membranes, escapes also into the spaces of Disse, to be consequently transported by the sinusoidal blood.

Acute Disease↗

Bile duct bacterial isolates in primary sclerosing cholangitis and certain other forms of cholestasis--a study of bile cultures from ERCP.

BACKGROUND/AIMS: The pathogenesis of the inflammatory lesion in primary sclerosing cholangitis is unknown. We have recently demonstrated a high positivity rate of bacterial cultures in bile and bile ducts of explanted livers from primary sclerosing cholangitis patients compared with patients with primary biliary cirrhosis. In particular, alpha-hemolytic Streptococci was a frequent finding, suggesting an etiopathogenic role of that particular bacteria in primary sclerosing cholangitis. We therefore wanted to study naive primary sclerosing cholangitis patients and compare them with primary sclerosing cholangitis patients that have previously undergone endoscopic retrograde cholangiopancreatography, in order to evaluate the potential role of these bacteria in the etiopathogenesis in primary sclerosing cholangitis. METHODOLOGY: Samples for bacterial cultures were obtained during a diagnostic endoscopic retrograde cholangiopancreatography. PARTICIPANTS: 12 naive primary sclerosing cholangitis patients, 10 patients with primary sclerosing cholangitis, previously investigated using endoscopic retrograde cholangiopancreatography, 47 patients with choledocholithiasis, 19 patients with cancer obstructing the common bile duct, and 29 patients with other forms of biliary disorders. RESULTS: Positive cultures were obtained from 3 of the naive primary sclerosing cholangitis patients and from 6 of the primary sclerosing cholangitis patients with previous endoscopic retrograde cholangiopancreatography (NS). The most frequent finding in all the primary sclerosing cholangitis patients was alpha-hemolytic Streptococci. Bacteria were cultured from the bile in 64% of the patients with choledocholithiasis, higher than the 25% in the naive primary sclerosing cholangitis patients (P < 0.03), and in 56% of patients with obstructing cancer (NS) but in only 24% of patients with other forms of biliary disorders, all of whom, except 4, had normal cholangiograms. In the 22 patients with primary sclerosing cholangitis, 75% of the positive bacterial cultures consisted of Gram-positive isolates and 25% were enteric bacteria, which differed statistically from the 74% enteric bacteria and 26% Gram-positive bacteria in the patients with common duct stone (P < 0.01). CONCLUSIONS: Alpha-hemolytic Streptococci do not seem to play a primary role in the etiopathogenesis of primary sclerosing cholangitis since most naive primary sclerosing cholangitis patients were found to have negative bacterial cultures. This does not exclude the possibility that they play a role in the progression of primary sclerosing cholangitis following infection in conjunction with the first endoscopic retrograde cholangiopancreatography.

Bacteria↗

[Effect of dalargin on the bile acid spectrum in human bile].

Effects were studied of dalargin on the bile acids spectrum in the bile of patients with duodenal ulcer. Dalargin has been shown to increase the bile concentration of tauro- and glycochilates, with the concentration of free bile acids remaining unchanged or being on the decrease.

Anti-Ulcer Agents↗

[Bacteriological study of bile from the gallbladder and bile ducts of patients surgically treated for biliary pathology].

We have evaluated the results of a bacteriological study of bile in 115 patients undergoing biliary surgery in the Valdivia Hospital (Chile). 35.6% of the positive bile cultures corresponded to patients with acute cholecystitis and only 22.5% to chronic disease. The culture was positive in 56.3% of cases of common bile duct stones and in 86.7% of cholangitis. Most positive bile cultures were monomicrobial, corresponding to patients with chronic cholecystitis. Most isolates were from Enterobacteriaceae, 49% of which corresponding to Escherichia coli. The in vitro sensitivity study showed that, as a rule, the strains were sensitive to the used antimicrobials.

Adult↗

[Selective resection of the distal bile duct in a high risk patient with polypoid cancer of the intrapancreatic bile duct].

An 81-year-old man who had suffered from hypertension for the preceding three years was admitted as an emergency to the department of neurology of our hospital with the chief complaint of dysarthria. He was diagnosed to have multiple lacunar cerebral infarcts by cranial CT, while the laboratory data showed liver dysfunction characteristic of cholestasis. Mild respiratory insufficiency and renal dysfunction were also found. Further radiological examinations on the liver and biliary tree (US, CT and ERC) were performed, and they revealed that the common bile duct was dilated due to two stone-like masses. He was referred to our department of surgery and underwent laparotomy. Intraoperative endoscopy disclosed intraductal papillomatous lesions. Because of the multiple complications of the patient, resection of the entire common bile duct including the gallbladder and the papilla of Vater without any resection of the pancreas was performed instead of pancreatoduodenectomy. The postoperative course was uneventful and he was discharged on the 31st POD. One year after operation, there is no sign of recurrence either clinically or radiologically. The potentially curative operation for cancer of the distal bile duct is pancreatoduodenectomy, but this is of so great a surgical stress that such a high risk case as described above might be unable to survive it. Even if he survived the perioperative period, he might have a poor quality of life due to postoperative complications. The macroscopic appearance of bile duct cancer is correlated to its invasive spread and prognosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenocarcinoma↗

[Cancer of the colon and cholelithiasis: bile bacteria, composition of the stones and the bile].

Our recent studies have shown a significant association between lithiasic biliary disease and colorectal cancer. This could be due to the existence of risk factors common to both disease or to a cause-effect correlation between them. This latter hypothesis is supported by the observation in gallstone patients of the increase of biliary and fecal concentrations in secondary biliary acids. These could have co-carcinogenic effect on the colon. With a view to singling out further elements which might help us to understand more clearly the possible cause-effect correlation between cholelithiasis and colon cancer, we examined 12 patients affected by both diseases. In these, we evaluated the composition of the gallbladder stones, by means of spectrophotometry and diffractometry. Bile samples were taken from the gallbladder and used to examine the lipidic composition and the cholesterol saturation index according to Carey. In addition bacteriological examinations were carried out. The results were compared with those of 10 patients with cholelithiasis but not cancer, 10 with cancer but not cholelithiasis and 10 with neither. Analysis of the results did not reveal significant differences in gallstone and bile composition between colon cancer patients with concomitant gallstones and control groups. However, in cancer patients with gallstones a higher incidence of bile bacteria (35.7%) was observed than in the other groups. Bile bacteria were observed more frequently in right colon cancer patients who had pigment stones in 75% of the cases. The results seem to evidence peculiarities in patients with a cancer of right colon.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Lipids, bile and fatty acids of the bile in rabbits with hypercholesterolemia].

Fractions of bile acids, neutral lipids, phospholipids as well as their fatty acid composition were studied in bile of 7 rabbits with alimentary hypercholesterolemia. Distinct alterations were observed in content of lipid fractions involved in cholesterol metabolism; the spectrum of fatty acids were transformed mainly towards increase in content of polyunsaturated acids, the bile acid spectrum was also altered. These alterations in metabolism of bile lipids appear to serve for neutralization of the unfavourable effect of cholesterol excess.

Animals↗

Determination of conjugated bile acids in human bile and duodenal fluid by reverse-phase high-performance liquid chromatography.

A simple mehtod using reverse-phase liquid chromatography is presented for resolution and quantitation of the major conjugated bile acids of man, including the glycine and taurine conjugates of the dihydroxy bile acids, chenodeoxycholic and deoxycholic acid. Using modern, high-performance chromatographic equipment, analysis time is less than 30 minutes. The quantitative range of the method, with detection by refractive index, is 0.05 to 0.1 mumol of bile acid and the limit of detection for an injection sample is 0.01 mumol. This provides a sensitivity sufficient for analysis of dilute duodenal and gallbladder bile with minimal sample preparation.

Bile↗

[Effect of bile and bile acids on bacterial resistance to antibiotics].

The combined effect of bile acids and antibiotics on the moderately sensitive and resistant staphylococcal strains was studied. 10 staphylococcal cultures isolated from patients with different pyo-inflammatory diseases were tested for their sensitivity to 5 antibiotics (benzylpenicillin, ampicillin, erythromycin, kanamycin and chlortetracycline). 9 of them were moderately sensitive to the drugs. The combined use of these antibiotics with cholic, desoxycholic , dehydrocholic, glycocholic and choleic acids increased the sensitivity of the staphylococci to the drugs in 87 per cent of the cultures. The same regularity was observed with respect to 15 staphylococcal strains isolated from the patients' bile. The combination of taurocholic acid and streptomycin was the most effective. The combinations of the antibiotics with cholic acid were less effective. The same regularities were noted in the experiments with billicultures tested for antibiotic sensitivity on solid media with the patient's bile. Antibiotics with maximum activity associated with the presence of the patient's bile should be recommended for the treatment of patients with inflammatory diseases of the liver and urinary tract.

Anti-Bacterial Agents↗

Bile restitution procedures for studying bile secretion in fistulated pigs.

In order to study bile secretion in the fistulated pig, an apparatus which reinfused the secreted bile at a rate reflecting the secretion rate, was used. The essential parts were a glass container equipped with two platinum wire electrodes, an electronic relay, a stop valve, and a peristaltic pump. This apparatus allowed a reinfusion of collected bile in a volume determined by the position of one of the electrodes, the recording of bile flow, and a continuous sampling for chemical analysis.

Animals↗

Bile acids. LXVII. The major bile acids of Varanus monitor.

The major bile acids of gall bladder bile of Varanus monitor have been separated by thin-layer chromatography and shown to be derivatives of taurine. After alkaline hydrolysis, the free acids were separated by thin-layer and partition chromatography. Identification or characterization of the free acids was facilitated by gas-liquid chromatography and gas-liquid chromatography-mass spectometry of the methyl esters or methyl ester-trimethylsilyl ethers. About 13% of the total bile acids was represented by the C24 acids cholic, deoxycholic, allocholic, chenodeoxycholic, and 12-oxo-3 alpha-hydroxy-5 beta-cholanic acids, of which cholic acid constituted about 50%. The remainder of the bile acids consisted of eight C27 acids of which varanic acid was the major constituent; an isomer of varanic acid and 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestanoic acid were also identified. By chromatographic behavior and mass spectral fragmentation, the structures of four C27 acids with unsaturated side chains were elucidated as follows: 3 alpha, 7 alpha-dihydroxy-5 beta-cholest-23-enoic, 3 alpha, 7 alpha-dihydroxy-5 beta-cholest-24-enoic, 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholest-23-enoic, and 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholest-24-enoic acids. Similarly, the structure of the 12-deoxy analog of varanic acid, 3 alpha, 7 alpha, 24 xi-trihydroxy-5 beta-cholestanoic acid, was suggested for the component that constituted 7% of the total.

Animals↗