Biomedical subjects
E W Moore
Publications and source records attributed to E W Moore.
A black hole: an unexpected cause of difficult intubation.
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Effect of NSAIDs on gallbladder bile composition.
Secretion of gallbladder mucin is an important step in gallstone pathogenesis. Previous studies have demonstrated that aspirin and other nonsteroidal antiinflammatory drugs (NSAIDs) can both inhibit gallbladder mucin secretion and prevent gallstone formation in animal models of cholesterol gallstone disease. The present study was performed to determine if chronic NSAID use was associated with a reduction in the mucin content or affected the lipid components of human gallbladder bile. Four groups of patients were identified retrospectively from a cohort of 230 morbidly obese patients who underwent gastric bypass surgery. The index group consisted of 18 patients who were found to have gallstones at gastric bypass surgery and had a history of chronic NSAID use. Three other patient groups were identified from the cohort by matching this index population for sex, race, and age according to the following criteria: (1) patients with gallstones who had not utilized NSAIDs, (2) patients without gallstones but with chronic NSAID use, and (3) patients without gallstones and without a history of NSAID use. Gallbladder bile was obtained from all patients by direct aspiration from the gallbladder at the time of surgery. Patients with gallstones had a significantly (P < 0.02) greater concentration of gallbladder mucin in their gallbladder bile compared to patients without gallstones (0.897 +/- 0.226 vs 0.173 +/- 0.039 mg/ml). Among gallstone patients, gallbladder mucin was reduced in those patients with a history of chronic NSAID use (1.18 +/- 0.43 vs 0.74 +/- 0.19 mg/ml).(ABSTRACT TRUNCATED AT 250 WORDS)
Some Ca2+ electrodes misrepresent Ca2+ binding by bile salts.
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Hypercalcemia decreases bile flow and increases biliary calcium in the prairie dog.
BACKGROUND: Biliary calcium is known to play an important role in the pathogenesis of gallstones. Calcium salts are present in all pigment gallstones and are also present in the core of most, if not all, cholesterol gallstones. METHODS: The effects of acute hypercalcemia on bile flow and biliary calcium secretion were examined in 22 prairie dogs during intravenous taurocholate infusion (0, 1.0, 2.25, and 4.5 mumol/kg/min). RESULTS: Bile flow was linearly correlated with bile acid output in both control (y = 7.62x + 13.5, r = 0.98) and hypercalcemic (y = 7.00x + 10.4, r = 0.96) animals. At lower bile acid outputs (< 3.0 mumol/kg/min), biliary ionized calcium output per increment bile acid output was significantly increased in hypercalcemic animals (0.016 versus 0.011 mumol Ca++ mumol taurocholate, p < 0.001). Bile ionized calcium concentrations approximated Gibbs-Donnan predicted values only at low bile flow rate. CONCLUSIONS: Hypercalcemia decreases bile flow and increases biliary ionized calcium concentration in the prairie dog. These effects favor the precipitation of calcium salts in bile.
Premicellar taurocholate enhances calcium uptake from all regions of rat small intestine.
BACKGROUND/AIMS: The specific components of bile, which is necessary for normal calcium absorption, are unknown. We have previously shown that Ca2+ is bound with high affinity by premicellar taurocholate. The current studies examined the effects of taurocholate on intestinal calcium transport. METHODS: Intestinal Ca2+ uptakes were measured from proximal, mid, and distal small intestinal segments perfused with solutions containing 45CaCl2 (0.1-1 mmol/L), taurocholate (0-10 mmol/L), trihydroxymethylaminomethane buffer (pH 7), phenolsulfonpthalein (nonabsorbable marker), and NaCl (total ionic strength, 0.16 mol/L) for four randomized perfusion periods. In other studies, the proximal small intestine was divided into two equal segments and perfused with either 45CaCl2 or 45CaCl2 plus taurocholate (2.5-5 mmol/L). Calcium absorption was measured from the difference in uptake and calcium concentration retained in mucosa. Finally, effects of taurocholate on Ca2+ uptake across isolated brush border membrane vesicles were measured. RESULTS: Premicellar taurocholate produced an approximately 1.7-2-fold enhancement (P < 0.01) in Ca2+ uptake in all regions, with lesser contributions from micellar taurocholate. These effects resulted in a net increase in calcium absorption. Premicellar taurocholate also significantly increased calcium uptake across brush border vesicles. CONCLUSIONS: Premicellar taurocholate significantly enhances calcium uptake into, and absorption across, enterocytes. The mechanisms remain to be experimentally verified.
Evidence that bile salts are important for iron absorption.
The role of bile salts in intestinal Fe absorption has not previously been defined. We have recently shown that bile salts having cholanic ring 7 alpha-OH and/or 12 alpha-OH groups bind Fe2+ with high affinity at premicellar concentrations and also produce a two-to threefold enhancement in Fe2+ uptake from perfused intestinal segments in vivo. However, the physiological relevance of these observations for Fe2+ absorption was not known. To study this, we examined the effects of a single dose of taurocholate (TC) administered with 59Fe-FeSO4 in reversing iron malabsorption induced by bile duct ligation. Fe absorption from three 0.5-mg doses of 59Fe-FeSO4 was measured from five groups of six adult Sprague-Dawley rats each: group 1, before and after bile duct ligation and subsequent administration of 59Fe-FeSO4 + TC test dose; group 2, before and after bile duct ligation followed by a third dose of Fe alone; group 3, before and after bile duct ligation followed by a third dose of Fe+taurodehydrocholate (TDHC); group 4, sham laparotomy; and group 5, nonoperated controls where Fe absorption was measured from three doses of 59Fe-FeSO4 at the same time intervals as group 1. Absorption was measured, using whole body counting, after achievement of steady-state counts after each Fe dose. Mean percentage absorption from dose 1 (baseline) was 35.33 +/- 4.6% (SE). No significant differences between any groups were noted. In nonoperated controls (group 4), Fe absorption was virtually identical after all three doses. Also, hematocrit did not change in any of the 30 animals.(ABSTRACT TRUNCATED AT 250 WORDS)
Gallbladder mucin, arachidonic acid, and bile lipids in patients who develop gallstones during weight reduction.
BACKGROUND: Arachidonic acid (AA) and hydrophobic bile salts (BS) stimulate gallbladder mucin (GBM) secretion, which is thought to be an essential step in gallstone pathogenesis. The present study was performed to evaluate the relationship between AA, BS, and GBM in patients who develop gallstones following weight reduction. METHODS: Eleven patients who underwent gastric bypass, developed symptomatic gallstones, and then underwent cholecystectomy were evaluated. Gallbladder bile was obtained for analysis during each procedure. Matched patients who did not develop gallstones following gastric bypass served as controls. RESULTS: GBM increased in every patient who developed stones (mean increase: 5000%). The largest increase was observed soon after gastric bypass, and this declined curvilinearly with time. Gallbladder bile cholesterol was initially elevated but then rapidly declined before increasing back to pregastric bypass levels after weight loss was complete. No significant changes in phosphatidylcholine molecular species (including AA) or BS composition were observed following weight reduction. Concentrations of cholesterol, phospholipids, and changes in [AA] over time were each a linear function of [BS]. No relationship between GBM and any of these bile constituents was apparent. CONCLUSIONS: These observations strongly suggest that increases in GBM, which occur with gallstone formation in humans, are not the result of alterations in biliary AA or BS composition.
Cultured human gallbladder epithelia. Methods and partial characterization of a carcinoma-derived model.
BACKGROUND: Gallstone disease remains a leading cause of morbidity and mortality in humans. Despite extensive research into the physiology of the gallbladder, little is known about mucosal events that precede and contribute to stone formation. Here, we describe and partially characterize a cultured epithelial model of human gallbladder mucosa. EXPERIMENTAL DESIGN: Cells originally obtained from a well-differentiated gallbladder mucosal carcinoma were cultured in modified Eagle's minimum media (supplemented with fetal calf serum and antibiotics) on polycarbonate supporting matrices. RESULTS: Cell cultures were observed to come to confluence with 6 to 9 days. Light and transmission electron microscopy demonstrated the resultant epithelia to be predominantly one cell thick, to be polar in orientation, and to have apical villi. Epithelia exhibited cytokeratin markers consistent with their epithelia origin, functionally acidified the mucosal bathing solutions, and secreted mucin. Further experiments demonstrated transepithelial potential differences, mucosal-to-serosal transfer of sodium which could be inhibited with amiloride and 4-acetamido-4'-isothiocyanatostilbene-2-2'-disulfonic acid, and paracellular movement of neutral molecular probes inversely related to size. CONCLUSIONS: This culture model of human gallbladder mucosal carcinoma cells exhibits parameters consistent with native gallbladder and may offer a convenient new research tool for the study of the pathophysiology of gallstone formation.
Gallstones in patients with morbid obesity. Relationship to body weight, weight loss and gallbladder bile cholesterol solubility.
Gallstones are common in obesity, and in individuals undergoing weight reduction. However, the relationships between body weight, weight reduction, gallbladder bile composition and gallstone formation are not well understood. The present studies were conducted on a cohort of 230 morbidly obese individuals presenting for bariatric surgery. Mean body weight ranged from 90-235.4 kg (mean: 136.2 kg). Body mass index (BMI) ranged from 35.4-94.7 kg/m2. Thirty-two patients (14%) had undergone prior cholecystectomy and 48 (21%) were found to have gallstones by intraoperative ultrasonography. No significant relationship was observed between gallstone prevalence and body weight. Following bariatric surgery weight loss averaged 1.57 kg/week over six months. Absolute weight loss ranged from 13.6-81.3 kg. Symptomatic gallstones requiring cholecystectomy developed in 15/150 patients (10%) over two years of follow-up. In contrast, ultrasonography detected asymptomatic gallstones in 34/92 patients (37%) six months following bariatric surgery. No relationship existed between the amount of weight lost and gallstone formation. Gallbladder bile cholesterol solubility remained constant throughout the entire weight range present in this population. No significant difference in cholesterol solubility was present between persons presenting for bariatric surgery and patients who developed symptomatic gallstones and underwent cholecystectomy following weight reduction. We conclude that gallstones are common in patients with severe obesity both before and following bariatric surgery. However, weight loss per se does not appear to be the major determinant of gallstone formation in persons who weigh in excess of 100 kg.
High-affinity binding is essential for enhancement of intestinal Fe2+ and Ca2+ uptake by bile salts.
Both calcium and iron are bound with high affinity by premicellar bile salts having cholanic ring 7-OH and/or 12-OH groups, forming soluble cation-bile salt complexes. The authors of the current study recently showed that premicellar taurocholate markedly enhances intestinal iron and calcium uptake. However, the relationship of high-affinity binding to the observed uptake enhancement was unknown. In the current study, this relationship was examined by studying taurodehydrocholate (TDHC) binding and intestinal uptake of both cations. Ca2+ binding was measured by noting depression of [Ca2+] activity in solutions containing constant total Ca concentrations (1 mmol/L) and varying [TDHC] (0.5-50 mmol/L). Fe2+ binding was assessed by equilibrium dialysis studies of 59FeSO4 (0.179-1.79 mmol/L) and TDHC (0.5-50 mmol/L). Effects of TDHC on intestinal Fe2+ and Ca2+ uptake were measured in isolated perfused intestinal segments in vivo in seven and eight Sprague-Dawley rats, respectively. TDHC, lacking ring OH groups, did not bind either cation with high affinity and had no effect on their intestinal uptake. These results suggest that high-affinity binding is essential for bile salt-induced enhancement of intestinal Fe2+ and Ca2+ uptake.
Changes in gallbladder bile composition following gallstone formation and weight reduction.
Changes in gallbladder bile composition that occurred in patients who developed gallstones during weight reduction were evaluated. Bile was sampled directly from the gallbladder in 11 morbidly obese patients with no gallstones at the time of gastric bypass surgery and after gallstones had formed at cholecystectomy. Bile salt concentration ([BS]) increased significantly from a mean of 82.7-157.7 mmol/L (P less than 0.05). The concentration of cholesterol in gallbladder bile increased slightly and cholesterol saturation declined slightly with weight reduction and gallstone formation. Gallbladder mucin concentration increased 18-fold from a mean of 62 to 1110 micrograms/mL (P less than 0.001). Both free [Ca2+] and total calcium [Ca] increased 40% from mean values of 1.12 and 5.05 mmol/L at gastric bypass to 1.86 and 8.60 mmol/L after gallstone formation (P less than 0.05). The increase in [Ca2+] observed after gallstone formation was much greater than anticipated from changes in [BS] alone. This excess [Ca2+] in gallbladder bile increased curvilinearly with increasing mucin concentration. These results show that both gallbladder mucin and [Ca2+] increase with gallstone formation in humans and that mucin may modulate [Ca2+] in gallbladder bile.
Convective movement of Ca2+ across guinea pig gallbladder epithelium.
Recently, much interest has developed in biliary calcium because of its importance in the pathogenesis and composition of gallstones. While much progress has been made in understanding the thermodynamic factors that control biliary calcium concentrations, little is known about the kinetic factors that control the movement of calcium across the gallbladder epithelium. These studies measure guinea pig gallbladder epithelial permeability to Ca2+ during in vivo convective water movement across the membrane. Water movement, ranging from -15.2 (absorption) to 6.3 microliters.min-1.cm-2 (water entry), was induced by placing hypotonic, isotonic, and hypertonic solutions into the gallbladder lumen. Calcium movement was found to be directly and linearly related to water flow, indicating that Ca2+ moved with the convective water flow, presumably across paracellular channels. The slope of this relationship (0.602), representing the concentration of calcium in the fluid translocated across the gallbladder epithelium, was only about half that of plasma or luminal contents, indicating that calcium movement across the membrane was restricted. The mean sieving coefficient (1 - r) of guinea pig gallbladder, calculated from this slope, was approximately 0.5, indicating that the epithelium is only moderately permeable to Ca2+. The results suggest that intraluminal chelation of Ca2+ for the possible prevention and/or treatment of calcium-containing gallstones is a potentially feasible therapeutic modality.
In vivo studies of biliary ceftriaxone excretion and solubility in guinea pig hepatic bile.
Ceftriaxone (CFTX), a third-generation cephalosporin, has occasionally been reported to produce biliary sludge composed of its calcium salt. We performed studies in guinea pigs to (1) investigate the hepatic route of CFTX excretion, (2) determine ceftriaione's effects on bile flow and composition, and (3) quantify the solubility and metastability of the calcium salt as a function of administered dose. Our results show that even at high doses ceftriaxone has only minimal effects on bile flow and biliary electrolyte secretion, either alone or in combination with bile salt (taurocholate) infusion. A significant increase in total calcium concentration was observed without change in free Ca2+ concentration, this is compatible with formation of a soluble calcium salt of ceftriaxone, as previously demonstrated in vitro. Ion products of Ca2+ and ceftriaxone as high as 3.5 times the solubility product constant without crystal formation were observed, confirming the presence of a metastable state for the calcium salt of ceftriaxone in the living animal. Biliary excretion of ceftriaxone inhibited excretion of indocyanine green, suggesting that ceftriaxone and indocyanine green share a common anionic excretory pathway in this species.
Calcium in human gallbladder bile.
Biliary calcium is believed to be of great importance in gallstone pathogenesis. These studies were therefore performed to determine if quantitative and/or qualitative differences in calcium are present in gallbladder bile from patients with and without gallstones. Bile was obtained by direct gallbladder aspiration from 68 obese patients undergoing elective gastric bypass surgery. Forty-five patients had no evidence of gallstones or sludge, 18 had cholesterol gallstones, and five had black pigment stones. Gallbladder bile was also obtained from 27 nonobese patients undergoing elective cholecystectomy (19 cholesterol; eight black pigment gallstones). For all patients, total calcium ranged from 1.50 to 16.44 mmol/L (mean: 6.05 +/- 0.31 mmol/L); free Ca++ ion ranged from 0.53 to 2.83 mmol/L (mean: 1.28 +/- 0.05 mmol/L). Considerable overlap was observed between obese and nonobese subjects and between patients with and without gallstones. For all patient groups, calcium, Ca++, and bound calcium increased linearly with increasing concentrations of bile salt. No significant differences in the slopes of these relationships were observed with obesity or gallstones. In contrast, free Ca++ ion was greater in gallbladder bile from gallstone patient groups throughout the entire range of bile salt. We hypothesize that this observed increase in Ca++ resulted from increased Gibbs-Donnan forces and excess gallbladder mucin present within the gallbladder bile of patients with gallstones.
Premicellar taurocholate enhances ferrous iron uptake from all regions of rat small intestine.
Inorganic iron is virtually insoluble at the pH of small intestinal contents. This severe solubility limitation has been partly overcome by intraluminal substances that bind and solubilize iron, thus increasing availability for absorption. While several dietary ligands capable of solubilizing Fe2+ in intestinal lumen have been described, an endogenous binding ligand has not been previously described. It has recently been shown that certain trihydroxy bile acids (taurocholate and glycocholate) show high-affinity premicellar and low-affinity micellar Fe(2+)-binding properties, resulting in the formation of soluble Fe(2+)-bile salt complexes. It was hypothesized that this binding would increase the intraluminal pool of soluble iron, increase delivery of soluble iron to mucosal carriers, and thus enhance intestinal Fe2+ uptake. As a first step toward testing this hypothesis, the effect of taurocholate on Fe2+ uptake from all regions of in vivo rat small intestine is reported. It is shown that taurocholate, at premicellar concentrations, produces a marked, stepwise increase in Fe2+ uptake from all regions of small bowel, with little further increase above the critical micellar concentration. Enhancement of intestinal Fe2+ uptake is a newly described effect, and potential physiological function, of premicellar bile salts.
Recovery of bile secretion following orthotopic liver transplantation.
Recovery of hepatic function following orthotopic liver transplantation includes the ability to produce 'adequate' bile. What constitutes adequate bile flow, however, has not previously been defined. The present study was undertaken to characterize biliary water and electrolyte secretion following hepatic transplantation. Bile was sampled from nine liver transplant recipients for 15-25 consecutive days during chronic t-tube biliary drainage. Liver biopsies and t-tube cholangiograms were unremarkable in all patients. During the first post-operative day mean bile flow, bile salt concentration, [BS], and bile salt output (BSO) were 60.0 microliters/min, 6.8 mM and 0.41 mumol/min, respectively. [BS] increased over days 1-5 and then plateaued at 12.2 mM over days 6-25 post-transplant. BSO and bile flow increased over days 1-12 before achieving steady-state values of 4.52 mumol/min and 334.7 microliters/min, respectively. In each patient bile flow increased linearly with increasing BSO. Choleretic index (CI), varied from 36.9-77.1 microliters/mumol (mean: 50.7 +/- 8.8). The y-intercept for this relationship ranged from 52.4-156.9 microliters/min (mean: 95.9 +/- 81.8). Only primary bile salts (82% cholate and 17% chenodeoxycholate), were observed in the bile of each patient. Biliary electrolyte concentrations were similar to that observed in plasma. Each was relatively unaffected by changes in bile flow and BSO. Electrolyte outputs increased linearly with respect to both BSO and bile flow. We conclude that recovery of bile secretion following orthotopic liver transplantation occurs gradually over a 10-12 day period and is strongly dependent upon bile salt secretion.
Canine bile contains anticrystallization factors that inhibit precipitation of calcium carbonate.
Previous studies have strongly suggested that human bile contains a substance(s) that interferes with the precipitation of calcium phosphate and carbonate from solution. These studies, however, did not distinguish between calcium binding by biliary constituents resulting in decreased calcium carbonate saturation (alterations in solution thermodynamics) and true inhibition of calcium salt precipitation by kinetic factors. Because our recent studies have shown that canine common duct bile is always supersaturated with calcium carbonate (thermodynamically at risk for precipitation), we hypothesized that it must contain kinetic factors that inhibit formation and/or growth of calcium carbonate crystals. Effects of canine bile, bovine albumin and the bile salt taurocholate on calcium carbonate precipitation were studied in highly supersaturated solutions of calcium carbonate that spontaneously undergo rapid precipitation. Measured free ionized calcium concentrations, [Ca++], and calculated calcium carbonate saturation indices were compared in test solutions and controls to evaluate the thermodynamic effects of test solutions on the degree of saturation in the assay system. It is shown that addition of only 0.2 ml of normal canine gallbladder bile to the assay system (a 1:101 dilution of biliary components) abolished precipitation. A lesser inhibitory effect (a decrease in the rate of precipitation) was observed when gallbladder bile was diluted but was lost after 10-fold dilution. Canine common duct bile caused a decrease in the rate of precipitation similar to diluted gallbladder bile. In contrast, sodium taurocholate (250 mmol/L), the major bile salt in the dog, and albumin (1.5 gm/L), the most abundant protein in bile, had only a minimal inhibitory effect.(ABSTRACT TRUNCATED AT 250 WORDS)