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These studies investigated the growth characteristics and functional properties of isolated canine pancreatic ductal epithelial cells. Cells were isolated from the accessory pancreatic duct and cultured by using three conditions: on vitrogen-coated petri dishes with fibroblast conditioned medium (nonpolarized); in vitrogen-coated Transwells above a fibroblast feeder layer (polarized); or as organotypic rafts above a fibroblast-embedded collagen layer (polarized). Growth characteristics, transepithelial resistances, and carbonic anhydrase and cyclic adenosine monophosphate (AMP) responses were evaluated. Under polarized conditions, the cells grew as monolayers with columnar epithelial characteristics. The monolayers developed high transepithelial resistance and became impervious to the passage of horseradish peroxidase. Epithelial growth factor (EGF) (2 ng/ml) stimulated ductal cell growth and accelerated the formation of a high-resistance monolayer. Forskolin (10 microM) rapidly decreased transepithelial resistance. Carbonic anhydrase activity, which was lower in nonpolarized compared with polarized conditions, was stimulated by carbachol (175 microM). Secretin, however, did not stimulate carbonic anhydrase activity in these cells. Although secretin stimulated adenylyl cyclase activity in early-passage cells, this response was lost in later-passage cells. Both vasoactive intestinal polypeptide (VIP; 1 microM) and forskolin (10 microM) consistently increased adenylyl cyclase activity. Isolated canine pancreatic ductal epithelial cells proliferate in vitro, develop high-resistance epithelial monolayers, and respond to stimuli that activate adenylyl cyclase. These cells should provide a useful model for regulatory studies of ductal cell functions.

Numerous investigators have proposed a role for bacteria in biliary lithogenesis. We hypothesized that bacterial DNA is present in gallstones, and that categorical differences exist between gallstone type and the frequency of bacterial sequences.Polymerase chain reaction (PCR) was used to amplify bacterial 16S rRNA and uidA (encoding Escherichia coli [E. coli] beta-glucuronidase) genes in different types of gallstones. PCR products were sequenced.Bacterial 16S rRNA and uidA DNA sequences in E. coli were detected in all brown pigment, common bile duct, and mixed cholesterol gallstones (n = 14). In contrast, only one (14%) of seven pure cholesterol gallstones yielded a PCR product. Most (88%) mixed cholesterol gallstones yielded PCR amplification products from their central, as well as their outer, portions. Sequenced products possessed 88-98% identity to 16S rRNA genes of E. coli and Pseudomonas species.Bacterial DNA sequences are usually present in mixed cholesterol (to 95% cholesterol content), brown pigment, and common bile duct, but rarely in pure cholesterol gallstones. The presence of bacterial beta-glucuronidase is also suggested. The role of bacteria and their products in the formation of mixed cholesterol gallstones, which comprise the majority of cholesterol gallstones, warrants further study.

Biliary infection is associated with mucin hypersecretion by the biliary epithelium. Mucins have been identified as potent pronucleators of cholesterol in bile. The aim of the present study was to determine whether lipopolysaccharides (LPS) from different bacteria are capable of stimulating mucin secretion by cultured dog gallbladder epithelial (DGBE) cells, and to investigate the mechanism by which LPS stimulate mucin secretion. Mucin secretion by confluent monolayers of DGBE cells was quantified by measuring the secretion of [3H]-N-acetyl-D-glucosamine-labeled glycoproteins. Cell viability was evaluated by measuring the leakage of the enzyme, lactate dehydrogenase (LDH), into the culture medium. LPS, derived from Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa (200 microg/mL), all caused an increase in mucin secretion by the DGBE cells, without causing concomitant cell lysis. LPS from E. coli was found to be the most potent stimulator of mucin secretion, and increased mucin secretion by the DGBE cells to 252% +/- 14% of control. LPS from E. coli had no effect on intracellular cyclic adenosine monophosphate (cAMP) levels in the DGBE cells. Addition of the nitric oxide (NO)-releasing compound, NOR-4 (0.125-1 mmol/L), to the cells did not result in increased mucin secretion, and the NO synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME) (4 or 10 mmol/L), did not inhibit the LPS-stimulated mucin secretion. Exogenous tumor necrosis factor alpha (TNF-alpha) (1-10 ng/mL) did cause a minor increase in mucin secretion by the DGBE cells, but the effect of LPS from E. coli on mucin secretion could not be inhibited by preincubation with a TNF-alpha antibody (10 microg/mL). We conclude that LPS stimulates mucin secretion by the gallbladder epithelium. Whether this stimulation is mediated by TNF-alpha remains to be determined.

We used confluent cultures of dog gallbladder epithelial cells, stimulated by conditioned medium from a culture of human neonatal foreskin fibroblasts, to establish the presence of inducible nitric oxide synthase (NOS, EC 1.14.13.39). Assay was by conversion of radiolabeled arginine to citrulline. By 4 days after addition of the conditioned medium, a relatively high level of activity was observed. However, further study showed that the enzyme did not require addition of the usual cofactors for maximal activity (NADPH, FAD, FMN and tetrahydrobiopterin) and was stable in the absence of anti-proteolytic agents. Our suspicion that this enzyme might not be NOS but arginine deiminase (EC 3.5.3.6) was confirmed by enzyme purification and by the liberation of ammonia during enzyme reaction. This enzyme, which is absent from primates and virtually confined to single-cell organisms, suggested the presence of Mycoplasma, a common contaminant of cell cultures, and it was subsequently confirmed that the fibroblast culture was a source of Mycoplasma. With the widespread interest in nitric oxide and NOS, and common use of the convenient [3H]arginine assay, there is a considerable danger of the two enzymes being confused. At the very least, it is necessary to check for activity in the absence of added cofactors.

We have successfully established monolayer and organotypic culture techniques for growing human oral and esophageal epithelial cells. Cells in monolayer culture were grown in serum-free medium, modified from techniques previously reported by our group. The organotypic cultures were grown in a defined medium supplemented with 10% fetal calf serum. Oral and esophageal cells were maintained in keratinocyte basal medium with pituitary extract and other supplements, and 0.05 mM calcium for 7-9 and 9-11 passages, respectively. Both cell types had similar morphology by phase contrast microscopy. When confluent, the cells were predominantly small, basaloid, and uniform and interspersed with larger, differentiated cells. By immunohistochemistry, both cell types in monolayer were positive to AE1, AE3, and 34BE12 antibodies to keratins of stratified epithelia. Oral epithelial cells in monolayer also were positive to 35BH11, representative of simple epithelial keratins, while esophageal cells were not. The esophageal cells were focally positive to K13, while the oral cells were negative. Both were negative for K19. When comparing monolayer to organotypic cultures and to in vivo specimens, there was a significant difference in the expression of keratins. Using organotypic cultures, AE1, AE3, and 34BE12 were strongly positive in both oral and esophageal cells, similar to in vivo tissues. In contrast to monolayers, both were also focally positive for K19. Esophageal cells were strongly positive for K13, while the oral cells were mildly but uniformly positive. Both were negative for keratins of simple epithelia. These two cell culture techniques offer unique opportunities to study the pathobiology, including carcinogenesis, of stable cell systems from the oral and esophageal epithelia.

1. Bile salts stimulate mucin secretion by the gallbladder epithelium. We have investigated whether this stimulatory effect is due to a detergent effect of bile salts. 2. The bile salts taurocholic acid (TC) and tauroursodeoxycholic acid (TUDC) and the detergents Triton X-100 (12.5–400 µM) and Tween-20 (0.1–3.2 mM) were applied to monolayers of cultured dog gallbladder epithelial cells. Mucin secretion was studied by measuring the secretion of [3H]N-acetyl-d-glucosamine-labelled glycoproteins. We also attempted to alter the fluidity of the apical membrane of the cells through extraction of cholesterol with β-cyclodextrin (2.5–15 mM). The effect on TUDC-induced mucin secretion was studied. Cell viability was assessed by measuring lactate dehydrogenase (LDH) leakage or 51Cr release. 3. In contrast with the bile salts, the detergents were not able to cause an increase in mucin secretion without causing concomitant cell lysis. Concentrations of detergent that increased mucin release (> 100 µM Triton X-100, > 0.8 mM Tween-20), caused increased LDH release. Incubation with β-cyclodextrin resulted in effective extraction of cholesterol without causing an increase in 51Cr release. However, no effect of the presumed altered membrane fluidity on TUDC (10 mM)-induced mucin secretion was observed. 4. The stimulatory effect of bile salts on mucin secretion by gallbladder epithelial cells is not affected by the fluidity of the apical membrane of the cells and also cannot be mimicked by other detergents. We conclude that the ability of bile salts to cause mucin secretion by the gallbladder epithelium is not determined by their detergent properties.

Attempts to grow human pancreatic duct epithelial cells in long-term culture have proven difficult. We have developed a system of growing these cells for several passages by adapting methods used to culture dog pancreatic duct cells. Epithelial cells were enzymatically dissociated from the main pancreatic duct and plated onto collagen-coated culture inserts suspended above a human fibroblast feeder layer. After primary culture, the cells were either passaged onto new inserts or plastic tissue culture plates in the absence of collagen. Cells grown on the latter plates were maintained in a serum-free medium. Primary pancreatic duct epithelial cells grow steadily to confluence as a monolayer in the feeder layer system. After primary culture, cells passaged onto new inserts with fresh feeder layer or plastic plates and fed with serum-free medium continued to develop into confluent monolayers for up to four passages. The cells were columnar with prominent apical microvilli, sub-apical secretory vesicles, and lateral intercellular junctions resembling the morphology of normal in vivo epithelial cells. These cells were also positive for cytokeratin 19, 7, and 8 and carbonic anhydrase II, as measured by immunohistochemistry. Metabolically, these cells synthesized and secreted mucin, as measured by incorporation of tritiated N-acetyl-D-glucosamine. In conclusion, we demonstrated that human pancreatic epithelial cells from the main duct can be successfully grown in culture and repeatedly passaged using a feeder layer system, with serum-free medium, and in organotypic cultures.

Cl- secretion by pancreatic duct epithelial cells (PDEC) regulates cellular HCO3- secretion, an important component of the exocrine pancreas. In cystic fibrosis, for example, impaired function of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel results in decreased pancreatic secretion and secondary pancreatic insufficiency. Studies of ion transport by PDEC have been hindered by the lack of a practical in vitro model. We have successfully cultured nontransformed dog PDEC on Vitrogen-coated permeable membranes overlying a feeder layer of myofibroblasts and report the characterization of Cl- channels in these cells. Cl- conductance, assessed through efflux of 125I from PDEC, was stimulated by agents acting via adenosine 3',5'-cyclic monophosphate (cAMP) or cytosolic Ca2+. The Cl- conductances activated by cAMP and Ca2+ were distinct, since they were differentially inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and, to a lesser extent, by 5-nitro-2-(3-phenylpropylamino)benzoic acid and diphenylamine-2 carboxylate. Patch-clamp studies confirmed the presence of Cl- channels activated by cAMP and Ca2+, with differential inhibition by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid. The presence of CFTR Cl- channels in PDEC was confirmed by immunoblotting. These cultured PDEC are an optimal model for studies of pancreatic duct secretion.

Lipids in hepatic bile may be modified by the gallbladder epithelium. The purpose of this study was to demonstrate a bidirectional exchange of cholesterol between biliary lipid carriers and gallbladder epithelial cells and to determine the factors regulating this cholesterol transfer. Gallbladder epithelial cells were cultured to confluent monolayers, their membranes were labeled with endogenously synthesized [14C]cholesterol, and the cells were incubated with model bile introduced into the apical membrane compartment. Similarly, model bile with different lipid composition containing [3H]cholesterol was incubated with the unlabeled monolayers. We found that cholesterol in the apical membrane bilayer of the epithelial cells exchanged readily with that in bile, but only in the presence of bile salts. The rate of exchange is dependent on the concentration and species of bile salts. The net gain of cholesterol (absorption) or net loss of cholesterol (cytotoxicity) exhibited by the epithelial cells was regulated by the thermodynamic stability of cholesterol and the detergent effect of mixed micelles in bile. It is also possible that the physicochemical composition of lipids in bile may modify the cellular function of the gallbladder epithelium.

To study the morphologic and genetic events associated with the carcinogenic process in the pancreas, we have isolated and cultured a cell line of dog pancreatic duct epithelial cells and treated these cells with a carcinogen. The pancreatic duct epithelial cells were plated onto Vitrogen-coated Transwell inserts suspended above a feeder layer of human gallbladder myofibroblasts. The epithelial cells grew steadily into polarized monolayers, could be passaged repeatedly, and demonstrated the typical morphologic, immunohistochemical, and flow cytometric profile of normal well-differentiated columnar pancreatic epithelial cells. After being treated with 10 -5 M N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for 48 h, the treated cells grew on plastic surfaces. When grown in organotypic culture, the MNNG-treated cells were cuboidal with a multilayered, pseudostratified architecture. Flow cytometry demonstrated aneuploidy and a high percentage of the cells in S phase after reaching confluency, in sharp contrast to untreated cells. Cytogenetic analysis of the MNNG-treated cells revealed frequent chromosomal trisomy and tetrasomy. The secretion of mucin was also different in the MNNG-treated cells versus the untreated cells. The cultured pancreatic epithelial cells may be useful as an assay system to study the genotoxicity of known and potential carcinogens.

Background: Sentinel lymph node (SLN) biopsy is a standard surgery for axillary staging of early breast cancer. This study aimed to evaluate the identification rate and surgery time of SLN biopsy with the multi-modal method using the mixture of indocyanine green (ICG) fluorescence, RI and blue dye as a new method, in comparison with the RI, in clinically node negative breast cancer patients. Materials and methods: In this phase-II randomized study, 86 patients with clinically node negative breast cancer were randomized to receive periareolar injection of the mixture including ICG fluorescence, RI and blue dye or the only RI for SLN biopsy. We compared the identification rate, the number of SLN, and detections time of SLN biopsy between these two groups. Results: The mean age of the multimodal group and the only RI group was 48.2 years and 51.0 years (p = 0.12), respectively. There were no differences of histopathologic factors including tumor size, node positivity, hormone receptor positivity and HER2 positivity between two groups. Sentinel lymph nodes were identified in all patients of both groups (100% in the multimodal group vs 100% in the RI group). The average number of SLN in the mixture group was more than that of the RI group (3.4 ±1.37 vs 2.3 ± 1.04; p < 0.001). The time to excise the first SLN was similar in each group (6.5 ± 5.16 min vs 8.0 ± 4.35 min; p = 0.13). In multimodal group, percutaneous lymphatic drainage was visualized by fluorescent imaging in 90.7% (39 of 43 patients) and all of the first SLN were detected by both ICG fluorescent and RI. After operation, there was no complication including skin staining or necrosis. Conclusions: This study is the first randomize trial which compared the multimodal method using the mixture of ICG fluorescence, RI and blue dye and the conventional RI method for SLN biopsy. The multimodal method is an available method for SLN biopsy, which allows percutaneous visualization of lymphatic vessels and intraoperative lymph node detection. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-01-03.

We have analyzed the chemical composition of primary intrahepatic stones from 72 Korean patients. Two types of concretions have been identified: brown pigment (calcium bilirubinate) stones and black-colored mixed stones. Brown pigment stones were found in 68% of all cases and the remainder (32%) consisted of mixed stones. Intrahepatic mixed stones had mean cholesterol and bilirubin contents of 46.6% and 25.9%, respectively, whereas calcium bilirubinate stones had mean cholesterol and bilirubin contents of 14.1% and 43.6%, respectively. Intrahepatic mixed stones had a smooth black-colored surface and on cross section, exhibited a distinct outer shell surrounding an inner yellow, cholesterol-rich body. The finding of intrahepatic mixed stones with high cholesterol content suggests that primary hepatolithiasis may result from at least two different conditions or disorders and points to different approaches to their treatment.

Hypersecretion of gallbladder mucin has been proposed as a pathogenic factor in gallstone formation. We investigated whether mucin secretion is modulated by biliary constituents using normal, well-differentiated dog gallbladder epithelial cells. Model biles or bile salts were applied to monolayers of epithelial cells. Mucin secretion was studied by measuring the secretion of [3H]N-acetyl-D-glucosamine-labeled glycoproteins. Model biles with different cholesterol saturation indices increased mucin secretion by the cells to an average 251% after 5 hours of incubation (P < 0.01). Mucin secretion remained elevated during a 24-hour period, suggesting a sustained effect on mucin secretion. There was no relation between the cholesterol or phospholipid concentration and the extent of stimulation of mucin secretion. Taurocholate caused a dose-dependent increase in mucin secretion, suggesting that bile salt was the bile component responsible for the stimulatory effect. At a concentration of 0.5 mmol/L, only the more hydrophobic bile salts taurochenodeoxycholate and taurodeoxycholate, but not the hydrophylic bile salts taurocholate and tauroursodeoxycholate, stimulated mucin secretion (P < 0.01). Bile salts play an important role in the regulation of mucin secretion. A shift in the bile salt composition of bile towards the more hydrophobic bile salts may cause mucin hypersecretion, thereby initiating cholesterol gallstone formation

Using a system of phosphatidylcholine-cholesterol vesicles to model the vesicle phase of mammalian bile (1:1 molar ratio) we evaluated whether very small amounts of C. perfringens phospholipase C activity (0.5-6.5 nmol/min per ml) could lead to vesicle fusion, a precursor step for cholesterol precipitation in gallbladder bile. Quasielastic light scattering spectroscopy (QLS) was used to monitor vesicle growth and aggregation in model bile (0.89 mM total lipid) in the presence of phospholipase C. Vesicle growth over 2 h could be detected with phospholipase activity as little as 0.5 nmol/min per ml. Vesicle growth was sustainable over days in the absence of Ca2+ once as little as 3-7 mol% diacylglycerol had been generated as a result of the initial phospholipase C treatment. The presence of fusion intermediates was confirmed using transmission electron microscopy. In addition, kinetically slow vesicle fusion with intravesicle content mixing and minimal leakage was also confirmed by fluorescence spectroscopy using two populations of vesicles containing 5 mM TbCl3 or 50 mM dipicolinic acid. Efficient fusion (40% maximum fluorescence) was obtained at 30 min at 25 degrees C with phospholipase C activity. This level of enzyme activity approximates that found in human gallbladder bile (1.2 nmol/min per ml). We conclude that the hydrolysis products of phospholipase C activity can, in very small amounts (3-7 mol% diacylglycerol), lead to destabilization and fusion of cholesterol-saturated biliary vesicles. A reappraisal of the importance of phospholipase C hydrolysis products in the pathogenesis of cholesterol gallstones is warranted based on these observations.

The dynamic interchange of cholesterol and the phase transition between nonmicellar and micellar aggregates in rat and model bile were characterized with gel-permeation chromatography, quasi-elastic light scattering, turbidity measurements, and by radiolabeling lipid aggregates in bile. Cholesterol partitioned into either the micellar or nonmicellar phases independent of the lipid aggregate structure. In model bile, increasing bile salt concentrations led to a decrease in the relative proportion of nonmicellar aggregates beginning at 5 mM taurocholate (TC), while the relative cholesterol content of the nonmicellar fraction increased from 1.0 to 2.7 +/- 2.0 (means +/- SD). In rats, creation of a biliary fistula resulted in a decrease of bile salts from 41 to 4 mM. Mixed micelles increased from 25 to 120 A in radius, while nonmicellar aggregates increased from 180 to 800 A in radius. Addition of TC to model bile (cholesterol:lecithin = 1:1) vesicles with total lipid concentrations less than 7 mM yielded a progressive shift of vesicles (450 A) to mixed micelles (30 A). For mixtures with higher total lipid concentrations, addition of TC promoted substantial vesicle aggregation and resulted in formation of a third phase containing lipid aggregates larger in size than the initial vesicles. These results suggest that rapid exchange of cholesterol occurs in bile and that significant remodeling of vesicles can occur. These alterations in vesicles include both enrichment in cholesterol content and formation of larger aggregates during increases in bile salt concentration.

Background. Cancers of the pancreas originate from both the endocrine and exocrine elements of the organ, and represent a major cause of cancer-related death. This study provides a comprehensive assessment of gene expression for pancreatic tumors, the normal pancreas, and nonneoplastic pancreatic disease. Methods/Results. DNA microarrays were used to assess the gene expression for surgically derived pancreatic adenocarcinomas, islet cell tumors, and mesenchymal tumors. The addition of normal pancreata, isolated islets, isolated pancreatic ducts, and pancreatic adenocarcinoma cell lines enhanced subsequent analysis by increasing the diversity in gene expression profiles obtained. Exocrine, endocrine, and mesenchymal tumors displayed unique gene expression profiles. Similarities in gene expression support the pancreatic duct as the origin of adenocarcinomas. In addition, genes highly expressed in other cancers and associated with specific signal transduction pathways were also found in pancreatic tumors. Conclusion. The scope of the present work was enhanced by the inclusion of publicly available datasets that encompass a wide spectrum of human tissues and enabled the identification of candidate genes that may serve diagnostic and therapeutic goals. © 2007 Lowe et al., published_or_final_version

The authors undertook a study to determine whether in vitro computed tomography (CT) or magnetic resonance (MR) imaging could enable the prediction of the outcome of gallstone dissolution with methyl tert-butyl ether (MTBE). In vitro CT and MR images were obtained of gallstones removed at surgery from 40 and 30 patients, respectively. The patterns of the gallstones seen on CT scans were categorized as dense, moderately dense, faint, isodense, rimmed, and laminated. Gallstones were categorized by maximal signal intensities seen on T1-weighted MR images. After imaging, gallstones underwent in vitro MTBE dissolution. CT appearances correlated well with dissolution rates. Greatest weight change was noted in gallstones with homogeneously faint and isodense patterns, and least weight change was seen in stones with a homogeneously dense pattern. Rimmed and laminated stones with foci of high attenuation dissolved significantly to 5% or less of the original weight, a finding indicating that such foci do not preclude dissolution. Dissolution rates correlated with attenuation values of homogeneous stones (r = .8) and of the rim portion of rimmed stones (r = .8). No correlation was found between T1-weighted signal intensities on MR images and MTBE dissolution rates.

Biliary sludge is a collection of mucus, calcium bilirubinate, and cholesterol crystals that is usually recognized by characteristic echoes on ultrasonography. Its pathogenesis, clinical significance, and ultimate prognosis remain uncertain. We therefore studied the origin of biliary sludge ultrasonic echoes, using an ex vivo liver-gallbladder preparation, and determined the outcome of a group of patients identified to have gallbladder sludge by ultrasonography. Echoes were not generated by either an increase in the total solid concentration or by the graded addition of partially purified mucus glycoprotein. Cholesterol monohydrate crystals (greater than 50 micron) mixed with mucus produced echoes that were indistinguishable from gallbladder sludge observed in patients. To determine the natural evolution of gallbladder sludge in patients, we prospectively followed 96 patients found to have biliary sludge for a mean of 37.8 mo by serial ultrasound scans every 6 mo. In 17 patients (17.7%) biliary sludge disappeared and did not recur for at least 2 yr, in 58 patients (60.4%) biliary sludge disappeared and reappeared, and in 8 patients (8.3%) asymptomatic gallstones developed. There were 12 cholecystectomies performed: six were done for symptomatic gallstones (6.3%) and the other six for sludge associated with severe biliary pain attacks with or without recurrent acute pancreatitis. The finding of sludge represented precipitates being formed in bile. In some patients, it was a precursor form of gallstone disease.

Peptic ulcer disease occurs when there is an imbalance between aggressive factors and mucosal resistance. Mucus plays a key role in mucosal resistance. The mucus layer is relatively resistant to pepti

Removal of the gallbladder results in an accelerated enterohepatic circulation of bile salts. It has been suggested that reflux of bile into the stomach after cholecystectomy may predispose a patient to the development of gastric lesions. To examine this hypothesis, 349 patients with endoscopically proved gastric ulcers or mucosal lesions were age and sex matched with 349 patients with upper gastrointestinal symptoms who had normal endoscopic findings. The frequency of cholecystectomy was determined to be 4.3 percent in the gastric ulcer group (control 3.7 percent) and 3.8 percent in the mucosal lesions group (control 4.2 percent). There was no statistical difference between patients with gastric ulcer and the control subjects, mucosal lesions and controls in the frequency of cholecystectomy using individually matched case-control comparison. The relative risk for gastric ulcer-matched control comparison was 1.3 (95 percent confidence limit 0.3 to 6.3). The relative risk for mucosal lesion-matched control comparison was 0.85 (95 percent confidence limit 0.2 to 2.9). The data did not support the hypothesis that cholecystectomy increases the propensity to develop gastric lesions.

When Dark Agouti rats were changed from a conventional pellet diet containing 3.3% crude fibre and 4% fat to the experimental diet containing 5% pectin and 17% fat, the levels of n-butyrate in the caecum rapidly decreased more than fivefold. Such a change could be important as n-butyrate is known to be a major energy source for colonocytes, and to affect colon tumour cell development in vitro. Striking decreases in the caecal concentrations of the short chain fatty acids isobutyrate, n-valerate and isovalerate occurred when Wistar rats were fed an experimental diet containing 5 % pectin as the only dietary fibre, compared to rats fed the experimental diet containing no dietary fibre. These large changes in the short chain fatty acid profile, caused by pectin supplementation, have important implications for the metabolism and health of the colon.

The interposition of a neutral starch gel greatly retarded bulk ionic movement by free flow. A mucus (charged) gel preparation of identical concentration and thickness further retarded ionic diffusion. The findings suggest that the charges in the mucus matrix may exert an ionic exclusion effect (Donnan Exclusion), thus retarding other ionic diffusion. We speculate that a mucus layer may physiologically behave as an ion exchange gel column and modify the traffic of charged ions through it.