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Bile
Kinds of Bile Terms modified by Bile Selected AbstractsBiological measurement of estrogenic activity in urine and bile conjugates with the in vitro ER-CALUX reporter gene assayENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2002Juliette Legler Abstract Although estrogens are excreted as biologically inactive conjugates, they can be reconverted to an active form, possibly by bacteria. A simple method was developed to deconjugate estrogen metabolites present in human urine and fish bile back to active estrogens by enzymatic hydrolysis with ,-glucuronidase or live Escherichia coli cells. Deconjugated extracts were tested for estrogenic activity in the in vitro stable estrogen receptor,mediated chemical-activated luciferase gene expression (ER-CALUX) assay. Estrogen glucuronides in urine obtained from human males and females were effectively converted to active forms after incubation with ,-glucuronidase or E. coli. The highest estrogenic activity was found in deconjugated metabolites from urine of a pregnant woman, in which levels up to 3,000 nmol estradiol equivalents per liter of urine were found after overnight incubation of urine with E. coli. Bile sampled from male bream and flounder from various freshwater and marine locations was also deconjugated and a good correlation was found between high biliary estrogenic activity and elevated levels of xenoestrogenic activity in surface water as well as in plasma vitellogenin. Therefore, the measurement of deconjugated bile could form a useful (indirect) biomarker for internal dose of xenoestrogens in male fish. [source] Multiple hepatic nodules: Rare manifestation of clonorchiasisJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 9 2006Wei-Chih Liao Abstract A 38-year-old man was admitted due to intermittent right upper quadrant pain for 1 month. Leukocytosis with marked eosionphilia and elevated serum alkaline phosphatase were noted. Stool examinations revealed no parasites or ova. Ultrasonography and computed tomography disclosed multiple hepatic tumors. Biopsy of the hepatic tumor was performed due to non-conclusive imaging studies and revealed eosinophil infiltration in portal areas only. Endoscopic retrograde cholangiography showed mild dilatation with irregularity of bilateral intrahepatic ducts, compatible with chronic cholangitis. Bile was aspirated and biliary lavage with normal saline was performed during endoscopy-guided biliary cannulation. Microscopic examination of the aspirate showed the characteristic ova of Clonorchis sinensis. The patient received Praziquantel therapy for 1 day. Abdominal pain reduced in intensity gradually. Eosinophilia and multiple hepatic lesions resolved after adequate treatment of Clonorchis sinensis. The rare manifestation of multiple hepatic tumors in Clonorchis sinensis should be differentiated from other primary or metastatic neoplasms, while biliary lavage for parasite ova is a valuable diagnostic tool when stool examination is negative. [source] Microbiologic Evaluation of Gallbladder Bile of Healthy Dogs and Dogs with Iatrogenic Hypercortisolism: A Pilot StudyJOURNAL OF VETERINARY INTERNAL MEDICINE, Issue 1 2010P.H. Kook Background: In people, hypercortisolism (HC) has been associated with acalculous cholecystitis and biliary dyskinesia, which may potentiate ascending biliary infections. In dogs, an association between HC and gallbladder disease recently has been documented, although the role of bacteria remains controversial. Furthermore, there is no information on the gallbladder bile microbial flora in healthy dogs. Objectives: To investigate the microbial flora in gallbladder bile in healthy dogs, the relationship between iatrogenic hyperadrenocorticism and bactibilia and possible changes in biliary microbial flora after cortisol withdrawal in dogs. Animals: Six control dogs and 6 dogs treated with hydrocortisone. Methods: Gallbladder bile obtained by percutaneous ultrasound-guided cholecystocentesis was cultured aerobically and anaerobically and examined cytologically before (d0), during (d28, d56, d84), and after (d28p, d56p, d84p) administration of hydrocortisone (8 mg/kg PO q12h). Results: In the control group, 2/42 bile cultures yielded bacterial growth (Enterococcus sp.; Escherichia coli on d0) and 1/42 bile smears had cytological evidence of bacteria (d28). In the HC group, 2/42 bile cultures yielded bacterial growth (Enterococcus sp. on d28; Bacillus sp. on d28p) and 3/42 bile smears had cytological evidence of bacteria (d84, d84, d28p). All dogs remained healthy throughout the study period (168d). Conclusions and Clinical Importance: Based on the results of conventional bacterial culture techniques, gallbladder bile of healthy dogs periodically may harbor bacteria, which do not appear to be clinically relevant. A 3-month period of iatrogenic HC was not associated with bactibilia. A higher prevalence of bactibilia may be detected with micromolecular techniques. [source] Bone mineral metabolism and histomorphometry in rats with cholestatic liver diseaseLIVER INTERNATIONAL, Issue 2 2002Zvi Ackerman Abstract: Background: The etiology of osteopenia in cholestatic liver disease is uncertain. An animal model is needed in order to study the efficacy of therapeutic agents. Aims: In order to characterise the bone disease in rats with cholestatic liver disease. Methods: Four-month old male Sprague,Dawley bile duct-ligated (BDL) and sham-operated (SO) rats were studied. Twenty-eight days after surgery serum osteocalcin, a bone-formation marker, urinary deoxypyridinoline (DPD) cross-links, a resorption marker, and 25-hydroxyvitamin D3 were determined. Static and dynamic (tetracycline-based) histomorphometric analysis was performed on femurs and tibiae. Results: All BDL rats developed biliary cirrhosis. Bile duct-ligated rats had lower bone mass, reflected in statistically significantly 13.5% lower femoral dry-weight, 16% lower femoral ash-weight, 42.7% lower tibial cancellous bone area and 19% lower trabecular thickness, compared with SO rats. Bile duct-ligated rats exhibited decreased bone formation manifested by statistically significantly 70% lower tetracycline double-labelling, 40% lower mineralising surface, 51% lower bone-formation rate and 47% lower osteocalcin compared with SO rats. Deoxypyridinoline levels were 20% lower in BDL rats. Bile duct-ligated rats had 52% lower serum 25-hydroxyvitamin D3 level, but no significant increase in cortical osteoid area. Conclusions: Bile duct-ligated rats develop osteopenia characterised by low bone-formation rate, and can be used for studying therapeutic agents for patients with cholestatic liver disease displaying similar bone changes. [source] Absence of glycochenodeoxycholic acid (GCDCA) in human bile is an indication of cholestasis: A 1H MRS studyNMR IN BIOMEDICINE, Issue 5 2009Omkar B. Ijare Abstract The utility of 1H MR spectroscopy in detecting chronic cholestasis has been investigated. The amide proton region of the 1H MR spectrum of human bile plays a major role in differentiating cholestatic (Ch) patterns from the normal ones. Bile obtained from normal bile ducts contains both taurine and glycine conjugates of bile acids , cholic acid (CA), chenodeoxycholic acid (CDCA), and deoxycholic acid (DCA). Absence of a glycine-conjugated bile acid glycochenodeoxycholic acid (GCDCA) has been observed in bile samples obtained from primary sclerosing cholangitis (PSC) patients. A total of 32 patients with various hepatobiliary diseases were included in the study. Twenty-one patients had PSC and 11 had normal cholangiograms. One PSC patient was excluded from the study because of a bad spectrum. Seventeen out of the 20 PSC patients showed an absence of GCDCA in their 1H MR spectrum of bile. Six of the 11 reference patients with normal cholangiogram also showed spectra similar to those of PSC, indicating the possibility of cholestasis. DQF-COSY and TOCSY experiments performed on bile samples from PSC patients also revealed absence of phosphatidylcholine (PC) in some of the bile samples, suggesting possible damage to the cholangiocytes by the toxic bile. These observations suggest that analysis of human bile by 1H MRS could be of value in the diagnosis of chronic Ch liver disorders. Copyright © 2008 John Wiley & Sons, Ltd. [source] Oesophageal and gastric bile exposure after gastroduodenal surgery with Henley's interposition or a Roux-en-Y loopBRITISH JOURNAL OF SURGERY (NOW INCLUDES EUROPEAN JOURNAL OF SURGERY), Issue 5 2004J.-Y. Mabrut Background: The degree which the various reconstruction techniques prevent bile reflux after gastroduodenal surgery has been poorly studied. Methods: Bile exposure in the intestinal tract just proximal to the jejunal loop was measured with the Bilitec 2000® device for 24 h after gastroduodenal surgery in three groups of patients. Group 1 comprised 24 patients with a 60-cm Henley's loop after total gastrectomy. Group 2 included 31 patients with a 60-cm Roux-en- loop after total (22 patients) or subtotal (nine) gastrectomy. Group 3 contained 21 patients with a 60-cm Roux-en- loop anastomosed to the proximal duodenum as part of a duodenal switch operation for pathological transpyloric duodenogastric reflux. Bile exposure, measured as the percentage time with bile absorbance greater than 0·25, was classified as nil, within the range of a control population of healthy subjects, or pathological (above the 95th percentile for the control population). Reflux symptoms were scored and all patients had upper gastrointestinal endoscopy. Results: Bile was detected in the intestine proximal to the loop in none of 24 patients in group 1, eight of 31 in group 2 and 12 of 21 in group 3 (P < 0·001). The mean reflux symptom score increased with the degree of bile exposure, and the proportion of patients with oesophagitis or gastritis correlated well with the extent of bile exposure (P < 0·001). Conclusion: A long Henley's loop was more effective in preventing bile reflux than a long Roux-en- loop. Bilitec® data correlated well with the severity of reflux symptoms and the presence of mucosal lesions. Copyright © 2004 British Journal of Surgery Society Ltd. Published by John Wiley & Sons, Ltd. [source] Pharmacological and Clinical Studies with Temocapril, an Angiotensin Converting Enzyme Inhibitor that is Excreted in the BileCARDIOVASCULAR THERAPEUTICS, Issue 3 2004Kenichi Yasunari ABSTRACT Temocapril is an angiotensin converting enzyme inhibitor (ACEI), a prodrug with a thiazepine ring. Its active form, temocaprilat, is slightly more potent than enalaprilat in inhibiting ACE isolated from rabbit lung. The inhibitory potency of temocaprilat on isolated rat aorta is 3 times that of enalaprilat. Temocapril is excreted in the bile and urine and can be used in patients with renal insufficiency. It reduces blood pressure without causing any significant change in heart rate or cardiac output. Temocapril has been reported to improve endothelial dysfunction in vitro by suppressing increased oxidative stress. In vivo it improves reactive hyperemia in patients with essential hypertension. It has been reported to prevent coronary vascular remodeling in vivo by suppressing local ACE and increased oxidative stress. In humans temocapril has been found to improve insulin resistance partly by increasing adiponectin levels. Cardiac remodeling was improved by temocapril not only in experiment animals but also in humans. It improves renal function and decreases urinary albumin excretion in diabetics as well as in hypertensive patients. Temocapril is currently marketed only in Japan. Considering its beneficial effects and unique pharmaco-kinetics, temocapril, is likely to be introduced in other countries as well. [source] Manganese cell labeling of murine hepatocytes using manganese(III)-transferrin,CONTRAST MEDIA & MOLECULAR IMAGING, Issue 3 2008Christopher H. Sotak Abstract Manganese(III)-transferrin [Mn(III),Tf] was investigated as a way to accomplish manganese-labeling of murine hepatocytes for MRI contrast. It is postulated that Mn(III),Tf can exploit the same transferrin-receptor-dependent and -independent metabolic pathways used by hepatocytes to transport the iron analog Fe(III),Tf. More specifically, it was investigated whether manganese delivered by transferrin could give MRI contrast in hepatocytes. Comparison of the T1 and T2 relaxation times of Mn(III),Tf and Fe(III),Tf over the same concentration range showed that the r1 relaxivities of the two metalloproteins are the same in vitro, with little contribution from paramagnetic enhancement. The degree of manganese cell labeling following incubation for 2,7,h in 31.5,µm Mn(III),Tf was comparable to that of hepatocytes incubated in 500,µm Mn2+ for 1,h. The intrinsic manganese tissue relaxivity between Mn(III),Tf-labeled and Mn2+ -labeled cells was found to be the same, consistent with Mn(III) being released from transferrin and reduced to Mn2+. For both treatment regimens, manganese uptake by hepatocytes appeared to saturate in the first 1,2,h of the incubation period and may explain why the efficiency of hepatocyte cell labeling by the two methods appeared to be comparable in spite of the ,16-fold difference in effective manganese concentration. Hepatocytes continuously released manganese, as detected by MRI, and this was the same for both Mn2+ - and Mn(III),Tf-labeled cells. Manganese release may be the result of normal hepatocyte function, much in the same way that hepatocytes excrete manganese into the bile in vivo. This approach exploits a biological process,namely receptor binding, endocytosis and endosomal acidification,to initiate the release of an MRI contrast agent, potentially conferring more specificity to the labeling process. The ubiquitous expression of transferrin receptors by eukaryotic cells should make Mn(III),Tf particularly useful for manganese labeling of a wide variety of cells both in culture and in vivo. Published in 2008 by John Wiley & Sons, Ltd. [source] ENDOSCOPIC TRANSPAPILLARY CATHETERIZATION INTO THE GALLBLADDER FOR DIAGNOSIS OF GALLBLADDER CARCINOMADIGESTIVE ENDOSCOPY, Issue 2 2006Naohito Uchida It is often difficult to determine the precise nature of lesions in the gallbladder by radiographic, endoscopic and ultrasonographic methods. The approach to the gallbladder by a percutaneous transhepatic route has been reported. However, there is a possibility of seeding tumor cells into the peritoneal cavity and liver in a percutaneous procedure. On the contrary, transpapillary route can be performed without a possibility of seeding. The double-contrast cholecystography, intragallbladder sonography, direct biopsy of gallbladder lesions and cytology using gallbladder bile have been performed by the procedure of the transpapillary catheterization into the gallbladder. Confirming malignancy by histopathological diagnosis is desirous for determining therapeutic strategy in gallbladder carcinoma. Gathering gallbladder bile is comparatively easier than biopsy of the lesion using the transpapillary catheterization into the gallbladder. Examination of telomerase-related molecules is useful for diagnosis of pancreatic carcinoma. Usefulness of combination assay of human telomerase reverse transcriptase mRNA (hTERT mRNA) and cytology using gallbladder bile obtained by transpapillary catheterization is reported here. However, it would appear that hTERT mRNA is less important in the diagnosis of gallbladder carcinoma than in that of pancreatic carcinoma. When the molecular biological substances with higher sensitivity are found, the reliance of the combination assay of the molecular biological substances and cytology will be established. [source] Newly Developed Ultrasonic Probe With Ropeway System for Transpapillary Intraductal Ultrasonography of the Bilio,Pancreatic Ductal SystemDIGESTIVE ENDOSCOPY, Issue 3 2000Naotaka Fujita Background: Intraductal ultrasonography of the bile/pancreatic duct using a thin-caliber ultrasonic probe (IDUS) provides excellent images of these ducts and the surrounding structures. Insertion of the device through the papilla of Vater is essential to carry out this examination. We developed a new probe with a ropeway system (XUM5RG-29R; Olympus, Tokyo) for transpapillary IDUS. Its usefulness such as ease of application and safety were prospectively evaluated. Patients and methods: During the period of October 1997 to April 1998, transpapillary IDUS using the probe was performed in 194 patients at seven medical institutions. The success rates of insertion of the probe into the bile/pancreatic duct, observation of the area of interest, and the incidence of complications were evaluated. Results: Passage of the probe through the papilla was successful without difficulty in all the patients. Successful introduction of the probe into the pancreatic duct, bile duct and both of the ducts was achieved in 98.4, 100 and 85% of the patients, respectively. Once the probe was introduced into the aimed duct, it was possible to obtain IDUS images of the area of interest in all but five patients. Mild acute pancreatitis developed in eight patients (4.1%), all of whom recovered with conservative therapy only. Conclusions: It is possible to introduce the new ultrasonic probe into the desired duct once a guide wire has been inserted. This type of ultrasonic probe is quite useful when performing transpapillary IDUS of the bile and/or pancreatic duct. [source] Whole stomach with antro-pyloric nerve preservation as an esophageal substitute: an original techniqueDISEASES OF THE ESOPHAGUS, Issue 2 2004J.-M. Collard SUMMARY., The paper describes an original technique of gastric tailoring in which the two-thirds of the lesser curvature proximal to the crow's foot are denuded flush with the gastric wall, leaving both nerves of Latarjet and the hepatic branches of the left vagus nerve intact. Maintenance of the vagal supply to the antro-pyloric segment in two patients resulted in the presence of peristaltic contractions sweeping over the antrum on simple observation of the antral wall at the end of the procedure and on both upper G-I series and intragastric manometry tracings 6 weeks postoperatively. Gastric exposure to bile on 24-h gastric bile monitoring was normal 6 weeks after the operation. Neither patient had any gastrointestinal symptoms with the exception of early sensations of postprandial fullness when overeating. [source] Estrogenicity in bile of juvenile rainbow trout as measure of exposure and potential effects of endocrine disruptorsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 5 2004Ann-Sofie Allard Abstract Estrogenicity in the bile of juvenile rainbow trout exposed to effluents from municipal sewage treatment plants and various industries was assayed by using a recombinant yeast strain containing the human estrogen receptor , gene. Estrogenicity in bile also was measured after deconjugation of steroids to provide an estimate of the exposure and as an endpoint for potential effects on the organism. In unexposed fish or fish exposed for three weeks at control localities, 0.5 to 9 ng of estradiol equivalents (EEq) were found per gram of bile (ng EEq/g bile). Fish exposed for three weeks in cages placed in the receiving waters near outlets of municipal effluent had an average activity of 26 ng EEq/g bile. Fish exposed to undiluted sewage water in aquaria had a bile estrogenicity of 51 to 87,000 ng EEq/g bile. Unconjugated estrogens contributed only 8% or less to the estrogenicity in bile of fish exposed to municipal effluents. Municipal sewage effluents were more estrogenic than the industrial effluents that were investigated. Estrogenicity in bile was compared to that in extracts of wastewater by using the same receptor assay, and to vitellogenin induction in the plasma of the same fish. Bile estrogenicity proved to be a useful and sensitive (internal) measure of exposure and indicated its potential for the display of biological effects as a complement or replacement of more laborious assays. [source] Biological measurement of estrogenic activity in urine and bile conjugates with the in vitro ER-CALUX reporter gene assayENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2002Juliette Legler Abstract Although estrogens are excreted as biologically inactive conjugates, they can be reconverted to an active form, possibly by bacteria. A simple method was developed to deconjugate estrogen metabolites present in human urine and fish bile back to active estrogens by enzymatic hydrolysis with ,-glucuronidase or live Escherichia coli cells. Deconjugated extracts were tested for estrogenic activity in the in vitro stable estrogen receptor,mediated chemical-activated luciferase gene expression (ER-CALUX) assay. Estrogen glucuronides in urine obtained from human males and females were effectively converted to active forms after incubation with ,-glucuronidase or E. coli. The highest estrogenic activity was found in deconjugated metabolites from urine of a pregnant woman, in which levels up to 3,000 nmol estradiol equivalents per liter of urine were found after overnight incubation of urine with E. coli. Bile sampled from male bream and flounder from various freshwater and marine locations was also deconjugated and a good correlation was found between high biliary estrogenic activity and elevated levels of xenoestrogenic activity in surface water as well as in plasma vitellogenin. Therefore, the measurement of deconjugated bile could form a useful (indirect) biomarker for internal dose of xenoestrogens in male fish. [source] Effect of bile salts, lipid, and humic acids on absorption of benzo[a]pyrene by isolated channel catfish (Ictalurus punctatus) intestine segmentsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 5 2001Lynn P. Weber Abstract Dietary absorption of lipophilic contaminants may be a significant route of exposure in aquatic organisms. Bile salts, lipids, and humic acids are important factors that may influence the intestinal absorption of a contaminant such as benzo[a]pyrene (BaP). We hypothesized that bile salts, monoglycerides, and free fatty acids would increase BaP intestinal absorption, while triglycerides, humic acids, and sediment would decrease BaP intestinal absorption. We have established and validated an in vitro model to examine modification of 3H-BaP absorption in everted intestinal segments from channel catfish (Ictalurus punctatus). Uptake of BaP into the everted intestinal segments continued to increase over the times examined in this study (60 min) and apparently occurs passively; thus, fugacity-based models of uptake are supported. Absorption of BaP into intestinal cells was significantly decreased by the addition of monoglycerides and free fatty acids to bile salts in the incubation media. Addition of triglycerides decreased BaP absorption even further. Humic acids may have decreased BaP intestinal absorption, while natural sediment may have increased BaP absorption. The results of this study suggest that all lipids may decrease intestinal uptake of lipophilic contaminants if they remain in unabsorbable excess in the intestinal lumen by retaining BaP in lipid/bile micelles. In contrast, if triglycerides are hydrolyzed into monoglycerides/free fatty acids prior to absorption, lipophilic contaminant uptake will likely be facilitated. Thus, it may be the hydrolytic state of lipids that determines its effects on BaP absorption. Humic acids alone may decrease dietary uptake of BaP, but our results suggest that other components in natural sediment may counteract this effect to cause a slight enhancement of BaP uptake. Further studies are needed to determine the dietary conditions necessary for bio-accumulation to contribute significantly to lipophilic contaminant body burdens in benthivorous fish. Finally, the everted intestinal segment technique has the potential to be used in other species and with different contaminants. [source] Using regional exposure criteria and upstream reference data to characterize spatial and temporal exposures to chemical contaminants,ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 4 2000Susan M. Cormier Abstract Analyses of biomarkers in fish were used to evaluate exposures among locations and across time. Two types of references were used for comparison, an upstream reference sample remote from known point sources and regional exposure criteria derived from a baseline of fish from reference sites throughout Ohio, USA. Liver, bile, and blood were sampled from white suckers (Catostomus commersoni) and common carp (Cyprinus carpio) collected during 1993 and 1996 in the Ottawa River near Lima, Ohio. Levels of exposure were measured for petroleum by naphthalene-type metabolites, combustion by-products by benzo[a]pyrene-type metabolites, coplanar organic compounds by ethoxyresorufin- O -deethylase (EROD) activity, and urea by blood urea nitrogen (BUN) levels. The four biomarkers analyzed proved effective in determining differences between reference and polluted sampling sites, between geographically close (<0.5 km) sites, and between sampling years at sites common in both years. Calculated exposure criteria levels of the polycyclic aromatic hydrocarbon bile metabolites were found to be a conservative approximation of levels from a designated reference site and could thereby permit comparison of biomarker levels of fish from the Ottawa River to a regional reference level. Polycyclic aromatic hydrocarbon bile metabolite and EROD activity levels were more reflective of spatial patterns of contamination than BUN, although all biomarkers indicated differences overtime. Biomarkers from white suckers seemed to be more responsive in detecting changes in contaminant levels than the same biomarkers from common carp. Lower levels in 1996 of all biomarkers at many sites suggested lower exposures than in 1993 and could be indicative of some improvement over the period. [source] Gallbladder Na+/H+ exchange activity is up-regulated prior to cholesterol crystal formationEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 8 2005S. C. Narins Abstract Background, Gallbladder Na+ and H2O absorption are increased prior to gallstone formation and may promote cholesterol nucleation. Na+/H+ exchange (NHE) isoforms NHE2 and NHE3 are involved in gallbladder Na+ transport in prairie dogs. We examined whether increased gallbladder Na+ absorption observed during early gallstone formation is the result of NHE up-regulation. Materials and methods, Native gallbladder and primary cultures of gallbladder epithelial cells (GBECs) harvested from prairie dogs fed nonlithogenic (CON) or 1·2% cholesterol diet for varying lengths of time to induce cholesterol-saturated bile (PreCRYS), cholesterol crystals (CRYS), or gallstones (GS) were used. NHE activity was assessed by measuring dimethylamiloride-inhibitable 22Na+ uptake under H+ gradient in primary GBECs. HOE-694 was used to determine NHE2 and NHE3 contributions. NHE protein and mRNA expression were examined by Western and Northern blots, respectively. Results, Gallbladder total NHE activity was 25·1 ± 1·3 nmol mg protein,1 min,1 in the control and increased during gallstone formation peaking at the PreCRYS stage (98·4 ± 3·9 nmol mg protein,1 min,1). There was a shift in NHE activity from NHE2 to NHE3 as the animals progressed from no stones through the PreCRYS and CRYS stages to gallstones. The increase in NHE activity was partly caused by an increased Vmax without any change in KNam. Both NHE2 and NHE3 protein increased moderately during the PreCRYS stage without increases in mRNA expression. Conclusions, Increased gallbladder Na+ absorption observed prior to crystal formation is in part caused by an increase NHE activity which is not fully accounted for by an increase in NHE proteins and mRNA levels but may be explained by enhanced localization in the membranes and/or altered regulation of NHE. [source] Effects of endothelin-1 on portal-systemic collaterals of common bile duct-ligated cirrhotic ratsEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 4 2004C.-C. Chan Abstract Background/Aims, Endothelin-1 (ET-1) may induce intrahepatic vasoconstriction and consequently increase portal pressure. Endothelin-1 has been shown to exert a direct vasoconstrictive effect on the collateral vessels in partially portal vein-ligated rats with a high degree of portal-systemic shunting. This study investigated the collateral vascular responses to ET-1, the receptors in mediation and the regulation of ET-1 action by nitric oxide and prostaglandin in cirrhotic rats with a relatively low degree of portal-systemic shunting. Methods, The portal-systemic collaterals of common bile duct-ligated (BDL) cirrhotic rats were tested by in situ perfusion. The concentration-response curves of collaterals to graded concentrations of ET-1 (10,10,10,7 m) with or without BQ-123 (ETA receptor antagonist, 2 × 10,6 m), BQ-788 (ETB receptor antagonist, 10,7 m) or both were recorded. In addition, the collateral responses to ET-1 with preincubation of N, -nitro-L-arginine (NNA, 10,4 M), indomethacin (INDO, 10,5 M) or in combination were assessed. Results, Endothelin-1 significantly increased the perfusion pressures of portal-systemic collaterals. The ET-1-induced constrictive effects were inhibited by BQ-123 or BQ-123 plus BQ-788 but not by BQ-788 alone. The inhibitory effect was greater in the combination group. Pretreatment of NNA or NNA plus INDO equivalently enhanced the response of ET-1 while pretreatment of INDO alone exerted no effect. Conclusion, Endothelin-1 has a direct vasoconstrictive effect on the collaterals of BDL cirrhotic rats, mainly mediated by ETA receptor. Endogenous nitric oxide may play an important role in modulating the effects of ET-1 in the portal-systemic collaterals of BDL cirrhotic rats. [source] Inhibition of prostacyclin by indomethacin ameliorates the splanchnic hyposensitivity to glypressin in haemorrhage-transfused common bile duct-ligated ratsEUROPEAN JOURNAL OF CLINICAL INVESTIGATION, Issue 2 2001F.-Y. Lee Prostacyclin (PGI2) is an important contributor to the mediation of hyporeactivity to vasoconstrictors and the development of hyperdynamic circulation in portal hypertensive states. Inhibition of PGI2 synthesis in haemorrhage-transfused partially portal vein-ligated rats could ameliorate the splanchnic hyposensitivity to glypressin, a long-acting vasopressin analogue. This study investigated whether the hyposensitivity to glypressin also exists in rats with common bile duct ligation (BDL) and whether the inhibition of PGI2 synthesis by indomethacin could potentiate the portal-hypotensive effect of glypressin in bleeding BDL rats. Two series of BDL rats were used. Series 1 investigated the haemodynamic effects of low dose glypressin (0·07 mg kg,1) in BDL rats with or without bleeding by catheterization. In series 2, haemodynamic parameters were measured in stable or bleeding BDL rats that were receiving intravenously high dose glypressin (0·2 mg kg,1) or indomethacin (5 mg kg,1) followed by high dose glypressin. In rats with a hypotensive haemorrhage, 4·5 mL of blood was withdrawn and 50% of the withdrawn blood was reinfused before the administration of glypressin or indomethacin. Splanchnic hyposensitivity to glypressin was demonstrated in haemorrhage-transfused BDL rats receiving high, but not low, doses of glypressin. Indomethacin infusion did not cause significant systemic and portal haemodynamic changes in bleeding BDL rats (P > 0·05). The addition of indomethacin significantly enhanced the portal-hypotensive effects of glypressin (P < 0·05) and potentiated the increases in mean arterial pressure induced by glypressin infusion (P < 0·001) in bleeding BDL rats. Splanchnic hyposensitivity to glypressin observed in haemorrhage-transfused BDL rats could be ameliorated by the addition of indomethacin, suggesting a role of endogenous PGI2 in its pathophysiology. [source] Functional analysis of the rat bile salt export pump gene promoterFEBS JOURNAL, Issue 14 2002Regulation by bile acids, drugs, endogenous compounds The 5, flanking region of the bile salt export pump (Bsep) gene was systematically analysed to provide the basis for understanding the mechanisms which regulate Bsep transcription. In addition substrates and drugs were investigated for their ability to alter Bsep promoter activity. Bsep promoter function was restricted to hepatocyte derived HepG2 cells. The 5, deletional analysis revealed a biphasic shape of reporter gene activities, indicating a suppressive element between nucleotides ,800 and ,512. Two consensus sites for the farnesoid X receptor (FXR) were located at nucleotides ,473 and ,64. The latter was characterized as functionally active in bile acid-mediated feed-back regulation of Bsep transcription. Bsep promoter activity was reduced by rifampin and ,-estradiol. The anti-estrogen tamoxifen stimulated promoter activity. Dexamethasone, hydrocortisone and phenobarbital had no effect on Bsep promoter activity. In conclusion, the data suggest that transcriptional regulation of the Bsep gene can be modulated by a number of endogenous compounds and xenobiotics. FXR was a major regulatory factor, mediating bile acid feed-back stimulation of Bsep transcription. [source] Characterization of the PCR inhibitory effect of bile to optimize real-time PCR detection of Helicobacter speciesFEMS IMMUNOLOGY & MEDICAL MICROBIOLOGY, Issue 2 2005Waleed Abu Al-Soud Abstract The inhibitory effect of human and porcine bile samples to detect Helicobacter DNA was studied by adding different concentrations of bile samples to PCR mixtures of six thermostable DNA polymerases containing cagA specific primers and Helicobacter pylori DNA. PCR products were amplified by using the Rotorgene system and SYBR Green I. Among the six DNA polymerases tested, rTth had the lowest sensitivity to bile inhibitors, whereas Taq and Tfl had the highest sensitivity. Bile proteins did not inhibit AmpliTaq DNA polymerase, whereas the fraction containing mainly bile acids and their salts inhibited the amplification capacity of AmpliTaq. Heating human bile at 98 °C and adding casein and formamide to the reaction mixture reduced the PCR inhibitory effect of bile. Therefore, a pre-PCR treatment based on dilution and heating of bile, adding casein and formamide to the reaction mixture of rTth DNA polymerase was found efficient to amplify DNA directly in bile. [source] Survey of antibiotic resistance in an integrated marine aquaculture system under oxolinic acid treatmentFEMS MICROBIOLOGY ECOLOGY, Issue 3 2006Etienne Giraud Abstract The consequences of antibiotic use in aquatic integrated systems, which are based on trophic interactions between different cultured organisms and physical continuity through water, need to be examined. In this study, fish reared in a prototype marine integrated system were given an oxolinic acid treatment, during and after which the level of resistance to this quinolone antibiotic was monitored among vibrio populations from the digestive tracts of treated fish, co-cultured bivalves and sediments that were isolated on thiosulfate,citrate,bile,sucrose. Oxolinic acid minimum inhibitory concentration distributions obtained from replica plating of thiosulfate,citrate,bile,sucrose plates indicated that a selection towards oxolinic acid resistance had occurred in the intestines of fish under treatment. In contrast, and despite oxolinic acid concentrations higher than minimum inhibitory concentrations of susceptible bacteria, no clear evolution of resistance levels was detected either in bivalves or in sediments. [source] Herbal alternatives to bear bile: effects of Scutellaria baicalensis Georgi on IL-6 promoter and CYP3A4 activitiesFOCUS ON ALTERNATIVE AND COMPLEMENTARY THERAPIES AN EVIDENCE-BASED APPROACH, Issue 2006SS Appiah [source] Bile acid treatment alters hepatic disease and bile acid transport in peroxisome-deficient PEX2 Zellweger mice,HEPATOLOGY, Issue 4 2007Megan H. Keane The marked deficiency of peroxisomal organelle assembly in the PEX2,/, mouse model for Zellweger syndrome provides a unique opportunity to developmentally and biochemically characterize hepatic disease progression and bile acid products. The postnatal survival of homozygous mutants enabled us to evaluate the response to bile acid replenishment in this disease state. PEX2 mutant liver has severe but transient intrahepatic cholestasis that abates in the early postnatal period and progresses to steatohepatitis by postnatal day 36. We confirmed the expected reduction of mature C24 bile acids, accumulation of C27,bile acid intermediates, and low total bile acid level in liver and bile from these mutant mice. Treating the PEX2,/, mice with bile acids prolonged postnatal survival, alleviated intrahepatic cholestasis and intestinal malabsorption, reduced C27,bile acid intermediate production, and prevented older mutants from developing severe steatohepatitis. However, this therapy exacerbated the degree of hepatic steatosis and worsened the already severe mitochondrial and cellular damage in peroxisome-deficient liver. Both untreated and bile acid,fed PEX2,/, mice accumulated high levels of predominantly unconjugated bile acids in plasma because of altered expression of hepatocyte bile acid transporters. Significant amounts of unconjugated bile acids were also found in the liver and bile of PEX2 mutants, indicating a generalized defect in bile acid conjugation. Conclusion: Peroxisome deficiency widely disturbs bile acid homeostasis and hepatic functioning in mice, and the high sensitivity of the peroxisome-deficient liver to bile acid toxicity limits the effectiveness of bile acid therapy for preventing hepatic disease. (HEPATOLOGY 2007;45:982,997.) [source] Quantifying anomalous intestinal sterol uptake, lymphatic transport, and biliary secretion in Abcg8,/, mice,,HEPATOLOGY, Issue 4 2007Helen H. Wang Sitosterolemia is caused by mutations in either ABCG5 or ABCG8, but simultaneous mutations of these genes have never been observed. To explore whether ABCG8, the sterol efflux (hemi-)transporter, plays a major role in determining intestinal absorption efficiency and hepatic secretion rates of cholesterol and sitostanol, we performed direct measurements of the absorption and lymphatic transport of these sterols in mice with chronic biliary and lymphatic fistulae, as well as the transport rates of radiolabeled cholesterol and sitostanol from plasma high-density lipoprotein (HDL) into bile in male Abcg8,/, and wild-type mice. We observed that the absorption and lymphatic transport rates of radiolabeled cholesterol and sitostanol were increased by ,40% and ,500%, respectively, in Abcg8,/, mice in the setting of constant intraduodenal infusion of micellar taurocholate and lecithin. Both strains displayed identical intestinal Npc1l1 expression levels and small intestinal transit rates. After 45 minutes of intraduodenal infusion, acute intestinal uptake rates of trace [14C]cholesterol and [3H]sitostanol were essentially similar in both groups of mice with intact biliary secretion. Furthermore, in wild-type mice, mass transport rate of [3H]sitostanol from plasma HDL into bile was significantly faster than that of [14C]cholesterol; however, no [3H]sitostanol and only traces of [14C]cholesterol were detected in bile of Abcg8,/, mice. Conclusion: Deletion of the Abcg8 gene alone significantly increases the mass of intestinal cholesterol and sitostanol absorption and reduces but does not eliminate hepatic secretion of cholesterol. Moreover, the mutation has no influence on acute uptake of cholesterol and sitostanol by the enterocyte nor small intestinal transit time. (HEPATOLOGY 2007;45:998,1006.) [source] Bicarbonate-rich choleresis induced by secretin in normal rat is taurocholate-dependent and involves AE2 anion exchanger,HEPATOLOGY, Issue 2 2006Jesús M. Banales Canalicular bile is modified along bile ducts through reabsorptive and secretory processes regulated by nerves, bile salts, and hormones such as secretin. Secretin stimulates ductular cystic fibrosis transmembrane conductance regulator (CFTR),dependent Cl, efflux and subsequent biliary HCO3, secretion, possibly via Cl,/HCO3, anion exchange (AE). However, the contribution of secretin to bile regulation in the normal rat, the significance of choleretic bile salts in secretin effects, and the role of Cl,/HCO3, exchange in secretin-stimulated HCO3, secretion all remain unclear. Here, secretin was administered to normal rats with maintained bile acid pool via continuous taurocholate infusion. Bile flow and biliary HCO3, and Cl, excretion were monitored following intrabiliary retrograde fluxes of saline solutions with and without the Cl, channel inhibitor 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) or the Cl,/HCO3, exchange inhibitor 4,4,-diisothiocyanatostilbene-2,2,-disulfonic acid (DIDS). Secretin increased bile flow and biliary excretion of HCO3, and Cl,. Interestingly, secretin effects were not observed in the absence of taurocholate. Whereas secretin effects were all blocked by intrabiliary NPPB, DIDS only inhibited secretin-induced increases in bile flow and HCO3, excretion but not the increased Cl, excretion, revealing a role of biliary Cl,/HCO3, exchange in secretin-induced, bicarbonate-rich choleresis in normal rats. Finally, small hairpin RNA adenoviral constructs were used to demonstrate the involvement of the Na+ -independent anion exchanger 2 (AE2) through gene silencing in normal rat cholangiocytes. AE2 gene silencing caused a marked inhibition of unstimulated and secretin-stimulated Cl,/HCO3, exchange. In conclusion, maintenance of the bile acid pool is crucial for secretin to induce bicarbonate-rich choleresis in the normal rat and that this occurs via a chloride,bicarbonate exchange process consistent with AE2 function. (HEPATOLOGY 2006;43:266,275.) [source] Novel biotransformation and physiological properties of norursodeoxycholic acid in humans,,HEPATOLOGY, Issue 6 2005Alan F. Hofmann Experiments were performed in 2 volunteers to define the biotransformation and physiological properties of norursodeoxycholic acid (norUDCA), the C23 (C24 -nor) homolog of UDCA. To complement the in vivo studies, the biotransformation of norUDCA ex vivo using precision-cut human liver slices was also characterized. In the human studies, both a tracer dose given intravenously and a physiological dose (7.9 mmol, 3.0 g) given orally were excreted equally in bile and urine. By chromatography and mass spectrometry, the dominant biotransformation product of norUDCA in bile and urine was the C-23 ester glucuronide. Little N -acyl amidation (with glycine or taurine) occurred. The oral dose induced a sustained bicarbonate-rich hypercholeresis, with total bile flow averaging 20 ,L/kg/min, a rate extrapolating to 2 L/d. The increased bile flow was attributed to cholehepatic shunting of norUDCA as well to the lack of micelles in bile. Phospholipid and cholesterol secretion relative to bile acid secretion decreased during secretion of norUDCA and its metabolites, presumably also because of the absence of micelles in canalicular bile. When incubated with human liver slices, norUDCA was glucuronidated, whereas UDCA was conjugated with glycine or taurine. In conclusion, in humans, norUDCA is glucuronidated rather than amidated. In humans, but not animals, there is considerable renal elimination of the C-23 ester glucuronide, the dominant metabolite. NorUDCA ingestion induces a bicarbonate-rich hypercholeresis and evokes less phospholipid and cholesterol secretion into bile than UDCA. Molecules that undergo cholehepatic shunting should be powerful choleretics in humans. (HEPATOLOGY 2005;42:1391,1398.) [source] Peptide antibiotic human beta-defensin-1 and ,2 contribute to antimicrobial defense of the intrahepatic biliary treeHEPATOLOGY, Issue 4 2004Kenichi Harada Human beta-defensins (hBDs) are important antimicrobial peptides that contribute to innate immunity at mucosal surfaces. This study was undertaken to investigate the expression of hBD-1 and hBD-2 in intrahepatic biliary epithelial cells in specimens of human liver, and 4 cultured cell lines (2 consisting of biliary epithelial cells and 2 cholangiocarcinoma cells). In addition, hBD-1 and hBD-2 were assayed in specimens of bile. hBD-1 was nonspecifically expressed immunohistochemically in intrahepatic biliary epithelium and hepatocytes in all patients studied, but expression of hBD-2 was restricted to large intrahepatic bile ducts in 8 of 10 patients with extrahepatic biliary obstruction (EBO), 7 of 11 with hepatolithiasis, 1 of 6 with primary biliary cirrhosis (PBC), 1 of 5 with primary sclerosing cholangitis (PSC), 0 of 6 with chronic hepatitis C (CH-C), and 0 of 11 with normal hepatic histology. hBD-2 expression was evident in bile ducts exhibiting active inflammation. Serum C reactive protein levels correlated with biliary epithelial expression of hBD-2. Real-time PCR revealed that in all of 28 specimens of fresh liver, including specimens from patients with hepatolithiasis, PBC, PSC, CH-C and normal hepatic histology, hBD-1 messenger RNA was consistently expressed, whereas hBD-2 messenger RNA was selectively expressed in biliary epithelium of patients with hepatolithiasis. Immunobloting analysis revealed hBD-2 protein in bile in 1 of 3 patients with PSC, 1 of 3 with PBC, and each of 6 with hepatolithiasis; in contrast, hBD-1 was detectable in all bile samples examined. Four cultured biliary epithelial cell lines consistently expressed hBD-1; in contrast these cell lines did not express hBD-2 spontaneously but were induced to express hBD-2 by treatment with Eschericia coli, lipopolysaccharide, interleukin-1, or tumor necrosis factor-,. In conclusion, these findings suggest that in the intrahepatic biliary tree, hBD-2 is expressed in response to local infection and/or active inflammation, whereas hBD-1 may constitute a preexisting component of the biliary antimicrobial defense system. Supplementary material for this article can be found on the Hepatology website (http:/interscience.wley.com/jpages/0270,9139/suppmat/index.html). (Hepatology 2004;40:925-932). [source] Endogenous ursodeoxycholic acid and cholic acid in liver disease due to cystic fibrosisHEPATOLOGY, Issue 6 2004Jeffery L. Smith Focal biliary cirrhosis causes significant morbidity and mortality in cystic fibrosis (CF). Although the mechanisms of pathogenesis remain unclear, bile acids have been proposed as potential mediators of liver injury. This study examined bile acid composition in CF and assessed altered bile acid profiles to determine if they are associated with incidence and progression of liver injury in CF-associated liver disease (CFLD). Bile acid composition was determined by gas,liquid chromatography/mass spectrometry in bile, urine, and serum samples from 30 children with CFLD, 15 children with CF but without liver disease (CFnoLD), and 43 controls. Liver biopsies from 29 CFLD subjects were assessed histologically by grading for fibrosis stage, inflammation, and disruption of the limiting plate. A significantly greater proportion of endogenous biliary ursodeoxycholic acid (UDCA) was demonstrated in CFnoLD subjects vs. both CFLD subjects and controls (2.4- and 2.2-fold, respectively; ANOVA, P = .04), and a 3-4 fold elevation in endogenous serum UDCA concentration was observed in both CFLD subjects and CFnoLD subjects vs. controls (ANOVA, P < .05). In CFLD, there were significant correlations between serum cholic acid and hepatic fibrosis, inflammation, and limiting plate disruption as well as the ratio of serum cholic acid/chenodeoxycholic acid to hepatic fibrosis, inflammation, and limiting plate disruption. In conclusion, elevated endogenous UDCA in CFnoLD suggests a possible protective role against liver injury in these patients. The correlation between both cholic acid and cholic acid/chenodeoxycholic acid levels with histological liver injury and fibrosis progression suggests a potential monitoring role for these bile acids in CFLD. (HEPATOLOGY 2004;39:1673,1682.) [source] Gene delivery of Cu/Zn-superoxide dismutase improves graft function after transplantation of fatty livers in the ratHEPATOLOGY, Issue 6 2000Thorsten G. Lehmann Oxygen-derived free radicals play a central role in reperfusion injury after organ transplantation, and fatty livers are particularly susceptible. Endogenous radical scavengers such as superoxide dismutase (SOD) degrade these radicals; however, SOD is destroyed rapidly when given exogenously. Therefore, an adenoviral vector encoding the Cu/Zn-SOD gene (Ad.SOD1) was used here to test the hypothesis that organ injury would be reduced and survival increased in a rat model of transplantation of fatty livers. Donors received chow diet (untreated), high-fat diet, or ethanol-containing high-fat diet. Some of the ethanol-fed donors were infected either with the gene lacZ encoding bacterial ,-galactosidase (Ad.lacZ), or Ad.SOD1. After liver transplantation, SOD activity and protein expression in liver, survival, histopathology, release of transaminases, free radical adducts in bile, and activation of NF-,B, I,B kinase (IKK), Jun-N-terminal kinase (JNK), and TNF, were evaluated. Ad.SOD1 treatment increased survival dramatically, blunted transaminase release, and reduced necrosis and apoptosis significantly. Free radical adducts were increased two-fold in the ethanol group compared with untreated controls. Ad.SOD1 blunted this increase and reduced the activation of NF-,B. However, release of TNF, was not affected. Ad.SOD1 also blunted JNK activity after transplantation. This study shows that gene therapy with Ad.SOD1 protects marginal livers from failure after transplantation because of decreased oxygen radical production. Genetic modification of fatty livers using viral vectors represents a new approach to protect marginal grafts against primary nonfunction. [source] A missense mutation in FIC1 is associated with greenland familial cholestasisHEPATOLOGY, Issue 6 2000Leo W. J. Klomp Greenland familial cholestasis is a severe form of intrahepatic cholestasis described among indigenous Inuit families in Greenland. Patients present with jaundice, pruritus, bleeding episodes, and steatorrhea, and die in childhood due to end-stage liver disease. We investigated the possibility that Greenland familial cholestasis is caused by a mutation in FIC1, the gene defective in patients with progressive familial intrahepatic cholestasis type 1 and many cases of benign recurrent intrahepatic cholestasis. Using single-strand conformation polymorphism analysis and sequencing of the FIC1 exons, a missense mutation, 1660 G,A (D554N), was detected and was shown to segregate with the disease in Inuit patients from Greenland and Canada. Examination of liver specimens from 3 Inuit patients homozygous for this mutation revealed bland canalicular cholestasis and, on transmission electron microscopy, coarsely granular Byler bile, as previously described in patients with progressive familial intrahepatic cholestasis type 1. These data establish Greenland familial cholestasis as a form of progressive familial intrahepatic cholestasis type 1 and further underscore the importance of unimpeded FIC1 activity for normal bile formation. 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