sincalide and Cholelithiasis

The mechanisms responsible for the abnormalities of gallbladder emptying in patients with chronic acalculous gallbladder disease (AGD) have not been elucidated. This study was designed to determine whether a muscle defect could explain this gallbladder dysfunction.. Gallbladder contraction induced by a continuous intravenous cholecystokinin octapeptide (CCK-8) infusion was determined by ultrasonography in control subjects, patients with AGD, pigment stones, and cholesterol stones. Muscle cells were obtained by enzymatic digestion. (125)I-CCK-8 binding and [(35)S]guanosine triphosphate gamma S (GTP gamma S) binding studies were performed.. In vivo gallbladder contraction induced by CCK-8 was significantly lower in AGD (29.4%) and cholesterol stones (28.8%) than in pigment stones (59.8%) and normal controls (57.8%; P < 0.01). In vitro muscle cell contraction induced by CCK-8 was also lower in AGD than in pigment stones. It remained impaired in AGD after stimulation with the G-protein activators GTP gamma S and AlF(4) and with the second messenger 1,2-dioctanoyl-sn-glycerol. However, GTP gamma S binding induced by CCK-8 and vasoactive intestinal polypeptide and the binding capacity of CCK receptors were not different between AGD and pigment stones.. These findings suggest that there is a good correlation between in vivo and in vitro gallbladder response to CCK-8 in patients with AGD. Unlike those found in cholesterol stones, the muscle defects in AGD appear to reside in the contractile apparatus.

Topics: Cell Membrane; Cholelithiasis; Cholesterol; Chronic Disease; Colic; Diglycerides; Gallbladder; Gallbladder Diseases; Gallbladder Emptying; Guanosine 5'-O-(3-Thiotriphosphate); Humans; In Vitro Techniques; Iodine Radioisotopes; Muscle Contraction; Muscle Fibers, Skeletal; Muscle, Smooth; Receptors, Cholecystokinin; Sincalide; Sulfur Radioisotopes; Ultrasonography; Vasoactive Intestinal Peptide

Diagnosis and identification of patients with acalculous biliary pain, who would benefit from surgery, remains a significant clinical problem. The cholecystokinin (CCK) provocation test helps diagnosis, but lack of consistency limits its usefulness.. To characterize the response of gallbladder muscle strips, from patients with acalculous biliary pain, to hormonal and muscarinic stimulation and to compare these with strips from gallstone patients and normal controls.. Eleven patients with acalculous biliary pain were studied, 5 had a positive CCK test. Eight gallbladders from gallstone patients and 6 from partial hepatectomies were used for comparison.. Muscle strips from the body and neck of the gallbladder were suspended in organ baths and dose-response curves were constructed for CCK-8 and carbachol.. In the acalculous group the strips from the body were less sensitive to carbachol than those of the neck.. Since we found no differences in the CCK responses for the groups, it casts doubt over the effectiveness of the CCK test to diagnose acalculous biliary pain. Since carbachol sensitivity was different, it might be that a similar test using muscarinic stimulation would help in the diagnosis of this difficult group of patients.

Topics: Adult; Aged; Carbachol; Cholecystitis; Cholelithiasis; Dose-Response Relationship, Drug; Female; Gallbladder; Humans; In Vitro Techniques; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Pain; Reference Values; Sensitivity and Specificity; Sincalide; Statistics, Nonparametric

The main purposes of this study were to investigate the best parameter for describing gallbladder emptying and whether gallbladder bile emptying should be induced with a bolus injection or continuous infusion of cholecystokinin-octapeptide (CCK-8).. Gallbladder emptying was measured by dynamic cholescintigraphy. Twelve healthy subjects and six patients with gallstones were examined twice with CCK-8 infusion cholescintigraphy, 0.3 ng CCK-8 kg per min for 60 min under identical circumstances. Another six healthy subjects randomly received bolus injection (0.04 microgram/kg) and infusion of CCK-8 (0.3 ng/kg per min for 60 min), respectively, during cholescintigraphy on two separate occasions. The choice of bolus dose was based on recommendations from the CCK-8 manufacturer. The infusion dose was chosen to produce plasma CCK concentrations similar to postprandial plasma CCK levels.. A parameter of gallbladder emptying, mean ejection fraction (EF), was defined as 100% minus the area under the time-activity curve normalized to 100% and divided by the time interval from maximum to minimum counts per minute. This parameter proved superior to the well known parameters, EFmax. and EF30, in regard to reproducibility in healthy subjects. The slope of the regression line for the mean EF was 0.998 and the intercept value approximately 0% (p = 0.0001). The mean coefficient of variation was 4%. Apart from a higher mean coefficient of variation, similar reproducibility results were seen in the six patients. The measurements of EF30 in healthy subjects scattered more widely around the mean compared to the mean EF and EFmax, which indicates poorer ability to separate normal from abnormal gallbladder emptying. Intravenous bolus injection of CCK-8 resulted in incomplete gallbladder emptying with a mean EF value of 16% (s.d. 9%; range 7%-32%) compared to 49% (s.d. 7%; range 37%-57%) following CCK-8 infusion (p = 0.004). Abdominal discomfort was observed in all subjects after administration of the bolus injection, whereas no complaints were reported during infusion.. Mean EF is the best parameter for describing gallbladder emptying. Moreover, slow infusion of a physiological dose of CCK-8 is preferable to induce gallbladder emptying because it results in more complete emptying and has no side effects.

Topics: Adult; Case-Control Studies; Cholelithiasis; Female; Gallbladder; Gallbladder Emptying; Humans; Imino Acids; Infusions, Intravenous; Injections, Intravenous; Male; Middle Aged; Observer Variation; Organotechnetium Compounds; Radionuclide Imaging; Reproducibility of Results; Sincalide; Technetium Tc 99m Diethyl-iminodiacetic Acid

In the present study, we adopted real-time ultrasonography to investigate the effect of atropine on the gallbladder emptying induced by CCK-OP in the normal subjects, and the difference of gallbladder emptying induced by CCK-OP between the "silent" gallstone patients and the controls. The results showed that: (a) CCK-induced gallbladder emptying in normal subjects was inhibited from 88.7% +/- 5.5% without atropine to 43.4% +/- 9.4% with atropine (P < 0.001). (b) The fasting gallbladder volume in the "silent" gallstone patients (26.7 +/- 10.9cm3) was significantly larger than that in the control (19.2 +/- 7.3cm3) (P < 0.05) and gallbladder emptying in the "silent" gallstones patients (44.0% +/- 8.7%) was significantly lower than that in the control (53.4% +/- 6.4%) (P < 0.01). We concluded that, significantly inhibited CCK-induced gallbladder emptying and there was significantly impaired gallbladder emptying in the patients with gallstones.

Topics: Adult; Atropine; Cholecystokinin; Cholelithiasis; Female; Gallbladder; Gallbladder Emptying; Gastrointestinal Agents; Humans; Male; Middle Aged; Muscarinic Antagonists; Sincalide; Ultrasonography

Gallbladder motility was studied by ultrasound in 100 healthy adult volunteers and 150 gallstone patients, in a subgroup of whom gallstone burden, type and number, gallbladder histology and tensiometric responses of gallbladder strips to cholecystokinin octapeptide were evaluated. Patients were divided into contractors (n = 108) and hypocontractors (n = 42), according to their gallbladder motility pattern in vivo. Contractors showed slower gallbladder emptying and increased fasting and postprandial residual volumes, although the ejected amount of bile was greater than that of controls (20.2 +/- SEM 1.1 vs 16.0 +/- 0.7 ml; p < 0.001). In contrast, hypocontractors exhibited slower and less complete gallbladder emptying than controls with a reduction in the absolute amount of ejected bile. Although gallbladder wall inflammation was mild and comparable in specimens from both groups of patients, the thickness of the muscular layer was greater in hypocontractors than in contractors (1073 +/- 76 vs 745 +/- 75 microns, p < 0.01) and related inversely to postprandial ejected volume (r = -0.42; p < 0.03; n = 32) but positively to gallstone volume (r = 0.40; p < 0.03; n = 32). Compared to contractors, gall-bladder muscle strips of hypocontractors exhibited a decrease in frequency and amplitude of spontaneous contraction and in maximal stress and receptor sensitivity to cholecystokinin octapeptide (0.1 nM-1 microM). Postprandial gallbladder evaculation was unaffected by stone number, and by the presence or absence of stone calcification. Gallstone volume was larger in hypocontractors (19.4 +/- 3.0 ml vs 9.6 +/- 0.9 ml, p < 0.001) than contractors. The comparison of in vitro contractility patterns between cholesterol, mixed and pigment stone patients showed a more severe defect in patients with cholesterol and mixed stones than in those with pigment calculi. In conclusion, in gallstone patients: (i) gallbladder motor dysfunction manifests mainly with increased fasting and postprandial residual volumes in contractors and with markedly increased postprandial residual volumes and decreased gallbladder emptying in hypocontractors; (ii) gallbladder kinetics seem to be influenced by stone volume and cholesterol content of calculi but not stone number, calcification or mild chronic cholecystitis; (iii) a form of hypertrophic leiomyopathy is observed in gallstone patients with the most impaired gallbladder motor function.

Topics: Adult; Aged; Cholecystitis; Cholelithiasis; Cholesterol; Female; Gallbladder; Humans; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Sincalide; Ultrasonography

A 45-minute infusion of an octapeptide of cholecystokinin (Kinevac; Squibb Diagnostics, New Brunswick, NJ) was used to measure the gallbladder ejection fraction during cholescintigraphy in 40 normal volunteers. Cholecystokinin cholescintigraphy was shown to be a reproducible test. The maximum mean gallbladder ejection fraction occurred 15 minutes after cholecystokinin infusion and was 74.5% +/- 1.9% (mean +/- SEM). A gallbladder ejection fraction greater than 40% (mean -3SD) was arbitrarily defined to be normal. The gallbladder ejection fraction test was then used to identify patients with acalculous biliary symptoms who may respond to cholecystectomy. A total of 103 patients was tested; 21 had abnormal gallbladder ejection fractions and were randomized into two groups, cholecystectomy or no operation. These patients were followed up symptomatically at 3-month intervals for 13-54 months (mean, 34 months). Of the 11 patients who underwent cholecystectomy, 10 (91%) lost their symptoms and 1 improved. Of the 10 patients in the group that did not undergo surgery, all continued to be symptomatic, 2 of whom requested cholecystectomy after 13 and 24 months, respectively. Of the 13 gallbladders obtained from surgery, 12 showed evidence of chronic cholecystitis, muscle hypertrophy, and/or narrowed cystic duct. A normal gallbladder ejection fraction was recorded in 82 patients, and further treatment was left to the discretion of their referring clinician. On follow-up, 50 patients were asymptomatic and 10 were symptomatic without specific treatment of the biliary tract; 14 underwent cholecystectomy, 8 of whom were asymptomatic. Pathological abnormalities were recorded in 6 of the removed gallbladders. It is concluded that the gallbladder ejection fraction obtained after a 45-minute infusion of cholecystokinin during cholescintigraphy is a reproducible measure of gallbladder emptying, and that cholecystectomy alleviates the biliary-type pain of patients with a reduced gallbladder ejection fraction.

Topics: Adult; Cholecystectomy; Cholelithiasis; Female; Gallbladder; Gallbladder Diseases; Humans; Imino Acids; Male; Organotechnetium Compounds; Pain; Radionuclide Imaging; Sincalide; Technetium Tc 99m Diethyl-iminodiacetic Acid

Long term total parenteral nutrition (TPN) induces biliary sludge and formation of gallstones. Cholecystectomy is hazardous in these patients because of the underlying medical problems. Therefore, a randomized, double-blind controlled study was designed to test the hypothesis that daily administration of cholecystokinin-octapeptide (CCK-OP) prevents the formation of biliary sludge in humans receiving long term TPN. Adult patients receiving TPN for more than 21 consecutive days were studied. After randomization of 15 patients, the study was concluded because statistical significance was achieved. Eight patients received saline solution (placebo) intravenously and seven received CCK-OP (50 nanograms per kilogram) intravenously over a ten minute period daily. The groups were similar with respect to age, sex, diagnosis, liver function tests, amylase levels, total TPN time and time of study. All of the patients underwent weekly ultrasound studies. Volume and emptying studies of the gallbladder in response to the study drug were performed after one week. None of the patients receiving CCK-OP had sludge whereas five of eight of the patients receiving placebo had sludge (p less than 0.02). The results of emptying studies showed significant contraction of the gallbladder in those in the CCK-OP group but not in the placebo group. These data suggest that CCK-OP given intravenously daily prevents TPN induced stasis and sludge of the gallbladder. We conclude that CCK-OP should be used as routine prophylaxis against biliary sludge and formation of gallstones in patients receiving long term TPN.

Topics: Adult; Bile; Cholelithiasis; Cholestasis; Double-Blind Method; Humans; Middle Aged; Parenteral Nutrition, Total; Randomized Controlled Trials as Topic; Sincalide; Time Factors

To report a comparison of compounded and proprietary sincalide in the evaluation of gallbladder ejection fraction during hepatobiliary scintigraphy.. Two patients were referred to nuclear medicine with symptoms consistent with hepatobiliary dysfunction. Both underwent hepatobiliary scintigraphy to evaluate anatomic and physiologic tract patency of the hepatobiliary system. Compounded sincalide, an adjuvant pharmaceutical used to evaluate gallbladder ejection fraction, was infused during hepatobiliary scintigraphy, and gallbladder ejection fractions were 11% and 24%, respectively. Hepatobiliary scintigraphy was repeated on both patients 72 hours later with proprietary sincalide used as the adjuvant pharmaceutical. The gallbladder ejection fractions were 32% and 72%, respectively.. The use of sincalide to evaluate gallbladder ejection fraction in hepatobiliary scintigraphy is widely accepted in the surgical and nuclear medicine community. In late 2001, the sole manufacturer of sincalide announced indefinite unavailability of the product. Following the announcement, several compounding pharmacies began selling extemporaneously compounded sincalide as a replacement. Use of the compounded product has assumed therapeutic equivalence.. Significant differences in gallbladder ejection fraction between compounded sincalide and sincalide in our patients are likely due to the intrinsic variability in response to sincalide. Clinicians should be aware of this variability, as well as the potential effect of concomitant medications.

Topics: Adult; Chemistry, Pharmaceutical; Cholelithiasis; Gallbladder; Gastrointestinal Agents; Humans; Male; Middle Aged; Radionuclide Imaging; Sincalide; Tomography, X-Ray Computed

A series of our studies have shown that formation of cholesterol-supersaturated bile in patients with cholesterol gallstone disease is causatively related to decreased gallbladder contractility and mucin hypersecretion by the gallbladder. Supersaturated bile may modify the composition of gallbladder membranes so that the transduction of smooth muscle regulatory signals is impaired, and it may enhance the inflammation-induced mucin secretion by the gallbladder. To achieve a better understanding of the mechanism by which supersaturated bile impairs the contractility, we studied changes in the expression levels of gallbladder cholecystokinin (CCK-A) receptor messenger ribonucleic acid (mRNA) in prairie dogs fed a high-cholesterol diet. Levels of pathobiological determinants in arachidonate metabolism which are important for mucin secretion were also measured in their bile. Adult male prairie dogs were randomly assigned to receive either a semisynthetic diet (SSD) or an SSD plus 1.2% cholesterol (a high-cholesterol diet) for 2-, 4-, and 6-week periods. The contractile force in response to CCK-octapeptide (CCK-8) was measured by using gallbladder muscle strips. The mRNA levels of the CCK-A receptor were determined by reverse-transcription polymerase chain reaction (RT-PCR). Parallel to the increase in the cholesterol saturation index, the contractile responses to CCK-8 decreased in the animals fed a high-cholesterol diet for 4 weeks and markedly decreased in the animals with gallstone formation. However, in contrast to the decreased contractility, the steady-state mRNA levels of the gallbladder CCK-A receptor were significantly increased in the animals fed a high-cholesterol diet in comparison with the corresponding control animals. In the bile, a high-cholesterol diet caused an increase in the proportion of arachidonyl-phosphatidylcholine species, where phospholipase A(2) activity, prostaglandin E(2), and mucin concentrations were increased parallel to the feeding period. Up-regulation of the CCK-A receptor mRNA in the gallbladder of animals fed a high-cholesterol diet associated with decreased contractility may be due to an impairment of CCK signaling related to increased membrane cholesterol contents and its related reaction of biological compensation in order to increase the receptor concentration. The results of the present study suggest that in prairie dogs fed a high-cholesterol diet both a decrease in gallbladder contractility related to impairment of

Topics: Animals; Arachidonic Acid; Bile; Blotting, Northern; Body Weight; Cholelithiasis; Cholesterol; Cholesterol 7-alpha-Hydroxylase; Cholesterol, Dietary; Crystallization; Dinoprostone; Disease Models, Animal; Fatty Acids; Gallbladder; Gene Expression; Hydroxymethylglutaryl CoA Reductases; Lipids; Liver; Male; Microsomes, Liver; Mucins; Muscle Contraction; Muscles; Phosphatidylcholines; Phospholipases A; Receptor, Cholecystokinin A; Receptors, Cholecystokinin; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sciuridae; Sincalide

Human gallbladders with cholesterol stones (ChS) exhibit an impaired muscle contraction and relaxation and a lower CCK receptor-binding capacity compared with those with pigment stones (PS). This study was designed to determine whether there is an abnormal receptor-G protein coupling in human gallbladders with ChS using (35)S-labeled guanosine 5'-O-(3-thiotriphosphate) ([(35)S]GTPgammaS) binding, (125)I-labeled CCK-8 autoradiography, immunoblotting, and G protein quantitation. CCK and vasoactive intestinal peptide caused significant increases in [(35)S]GTPgammaS binding to Galpha(i-3) and G(s)alpha, respectively. The binding was lower in ChS than in PS (P < 0.01). The reduced [(35)S]GTPgammaS binding in ChS was normalized after the muscles were treated with cholesterol-free liposomes (P < 0.01). Autoradiography and immunoblots showed a decreased optical density (OD) for CCK receptors, an even lower OD value for receptor-G protein coupling, and a higher OD for uncoupled receptors or Galpha(i-3) protein in ChS compared with PS (P < 0.001). G protein quantitation also showed that there were no significant differences in the Galpha(i-3) and G(s)alpha content in ChS and PS. We conclude that, in addition to an impaired CCK receptor-binding capacity, there is a defect in receptor-G protein coupling in muscle cells from gallbladder with ChS. These changes may be normalized after removal of excess cholesterol from the plasma membrane.

Topics: Autoradiography; Cholecystokinin; Cholelithiasis; Cholesterol; Cross-Linking Reagents; Gallbladder; GTP-Binding Proteins; Guanosine 5'-O-(3-Thiotriphosphate); Humans; Immunoblotting; Immunosorbent Techniques; Iodine Radioisotopes; Muscle Contraction; Muscle, Smooth; Receptors, Cholecystokinin; Sincalide; Sulfur Radioisotopes; Vasoactive Intestinal Peptide

Iron deficiency has been demonstrated in the prairie dog to result in cholesterol crystal formation and altered biliary motility. Gallbladder filling and emptying are influenced by both inhibitory and excitatory stimuli, with nitric oxide (NO) playing a key role in normal relaxation. Iron is a cofactor for nitric oxide synthase. Therefore, we tested the hypothesis that iron deficiency would result in diminished levels of gallbladder neuronal nitric oxide synthase (nNOS) but would not influence the gallbladder's response to excitatory stimuli.. Twenty adult female prairie dogs were fed either an iron-supplemented (Fe(+)) (200 ppm) control diet (n = 10) or an iron-deficient (Fe-) (8 ppm) diet (n = 10) for 8 weeks. Fasting gallbladder volume was measured. Gallbladder muscle strips were harvested for response to excitatory stimuli and measurement of nNOS protein levels by Western blotting. Muscle strip response to a spectrum of doses of cholecystokinin, acetylcholine, and electrical field stimuli was determined, and the areas under the response curves were calculated.. Gallbladder volume increased in the iron-deficient prairie dogs compared with the iron-supplemented group (1.45 +/- 0.27 mL vs 0.80 +/- 0.13 mL, P < 0.05). Iron deficiency diminished the ratio of gallbladder nNOS to beta-actin protein levels (0.05 +/- 0.01 vs 3.48 +/- 1.02, P < 0.05) but resulted in a normal response to excitatory stimuli.. We conclude that diminished gallbladder neuronal nitric oxide synthase contributes to the gallbladder stasis that occurs with iron deficiency. This phenomenon may contribute to the increased incidence of gallstones in premenopausal women.

Topics: Animals; Blotting, Western; Body Weight; Cholelithiasis; Cholesterol; Dogs; Female; Gallbladder; Iron Deficiencies; Muscle Contraction; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Sincalide; Transferrin

Human gallbladders with cholesterol stones exhibit impaired muscle contraction induced by agonists that act on transmembrane receptors, increased membrane cholesterol content, and abnormal cholesterol-to-phospholipid ratio compared with those with pigment stones. The present study was designed to investigate the functions of the CCK receptor of gallbladder muscle membranes by radioreceptor assay and cross-linking. 125I-labeled CCK-8 binding was time-dependent, competitive, and specific. Scatchard analysis showed that the maximum specific binding (Bmax) was significantly decreased in cholesterol compared with pigment stone gallbladders (0.18 +/- 0. 07 vs. 0.38 +/- 0.05 pmol/mg protein, P < 0.05). In contrast, the affinity for CCK was higher in cholesterol than pigment stone gallbladders (0.18 +/- 0.06 vs. 1.2 +/- 0.23 nM). Similar results were observed in binding studies with the CCK-A receptor antagonist [3H]L-364,718. Cross-linking and saturation binding studies also showed significantly less CCK binding in gallbladders with cholesterol stones. These abnormalities were reversible after incubation with cholesterol-free liposomes. The Bmax increased (P < 0.01) and the dissociation constant decreased (P < 0.001) after incubation with cholesterol-free liposomes. In conclusion, human gallbladders with cholesterol stones have impaired CCK receptor binding compared with those with pigment stones. These changes are reversed by removal of the excess membrane cholesterol. These receptor alterations may contribute to the defective contractility of the gallbladder muscle in patients with cholesterol stones.

Topics: Bile Pigments; Cell Membrane; Cholelithiasis; Cholesterol; Cross-Linking Reagents; Devazepide; Gallbladder; Hormone Antagonists; Humans; In Vitro Techniques; Liposomes; Muscle, Smooth; Receptor, Cholecystokinin A; Receptors, Cholecystokinin; Sincalide

This study sought to determine whether there is a positive correlation between gallbladder emptying, biliary pain, and in vitro contractility.. Ultrasound measurements were carried out on 25 gallstone patients. The response of gallbladder strips to 1.75*10(-11) to 5.25*10(-7) M cholecystokinin-8 was recorded. In a second study 23 patients filled in pain questionnaires, and in vitro studies were again carried out.. Of five patients with no gallbladder emptying, four had in vitro contraction. Overall, a significant, positive linear correlation was found (P < 0.0001). In the second study in vitro contractility showed a positive linear correlation with pain.. Gallbladder emptying correlates with contractility. However, since most 'non-contractors' can contract, we suggest the term 'non-emptying' or 'emptying' to describe gallbladder dynamics. The positive correlation between pain and contractility suggests that biliary pain has a muscular component.

Topics: Cholelithiasis; Colic; Female; Gallbladder; Gallbladder Emptying; Humans; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Pain Measurement; Sincalide; Ultrasonography

The aim of this study was to determine the role of nitric oxide (NO), as the primary neurotransmitter of non-adrenergic, non-cholinergic (NANC) innervation, in stone-diseased and stone-free human gallbladders.. Human gallbladder muscle strips were mounted in modified Krebs-Henseleit solution with atropine (1 mM), guanethidine sulfate (5 mM) and aerated with Carbogen. Electric field stimulation (EFS) (70 V, 0.5 ms, 100 pulses) was used to activate NANC nerves. N-omega-nitro-L-arginine (L-NNA, 100 mM) and L-arginine (L-Arg, 120 mM) were used to manipulate the NO-synthase. Gallbladder slices were stained by using the alkaline phosphatase, anti-alkaline phosphatase (APAAP) method for histological examination.. In the control group (basal tone, 8.94 +/- 1.17 mN) caused a frequency-dependent reduction of basal tone (1 Hz = 5.73 +/- 0.81 mN; 3 Hz = 5.18 +/- 0.65 mN; 10 Hz = 4.63 +/- 0.49 mN), inhibition of NO-synthesis with L-NNA increased the tone (7.63 +/- 0.76 mN). Stone-diseased groups were divided into two groups (contractor and subcontractor), according to their ability to contract by CCK. Under the influence of EFS the contractor group (basal tone = 7.79 +/- 0.93 mN) reacted like the control group, but was frequency-independent and, additionally, showed spontaneous phasic contractions. In the sub-contractor group (basal tone 4.13 +/- 0.65 mN) EFS only decreased the frequency of spontaneous phasic contractions. L-NNA caused an increase in tone (5.97 +/- 0.84 mN) and frequency. L-arginine, the substrate for NO-synthase, significantly reversed this effect, indicating the dependence on NO. Histologically, the contractor group showed a wrinkled mucosal membrane and low-grade inflammation. Shallow mucosa, necrosis and high-grade inflammation were found in the sub-contractor group.. In vitro, NANC relaxation of human gallbladder is NO dependent. The motility of stone-diseased gallbladders is modulated by NO and seems to depend on the degree of scarification.

Topics: Adult; Aged; Cholelithiasis; Electric Stimulation; Enzyme Inhibitors; Female; Gallbladder; Gastrointestinal Agents; Humans; In Vitro Techniques; Male; Middle Aged; Muscle Contraction; Nitric Oxide; Nitric Oxide Synthase; Nitroarginine; Receptor, Cholecystokinin A; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; Sincalide; Tetrodotoxin

To compare the common procedure in tensiometry of normalization of the force (in N) produced by a gallbladder tissue strip to units of stress, with normalization of force to the strip content of contractile protein.. A comparison was made in both healthy and in diseased gallbladder tissue strips between two normalization procedures involving anatomical parameters. The contractile response expressed in terms of tissue stress (in N/m2), which entails a normalization to the strip cross-sectional area, was set against normalization to the tissue content of contractile protein (in N/mg actin/g strip wet weight).. Dose-response curves for acetylcholine (ACh) (10(-8) to 10(-3) M) and sulphated cholecystokinin octapeptide (CCK) (10(-12) to 10(-6) M) were assessed in healthy guinea pig (n = 8) and in diseased human gallbladder tissue strips (n = 28). Assuming a tissue density of 1.05 g/cm3, the strip cross-sectional area was calculated from its weight and length. Actin content in homogenized strips was determined by polyacrylamide gel electrophoresis followed by densitometry.. Actin content in human tissue was 19.06 +/- 1.42 mg/g strip wet weight, and 12.84 +/- 0.76 mg/g strip wet weight in guinea pig tissue. No correlation was found between strip cross-sectional area and actin content. In the diseased human tissue, no correlation was found between the inflammation score and either strip cross-sectional area or strip actin content. Maximal force (in mN) exerted in response to either ACh or CCK correlated much more closely in healthy guinea pig gallbladder (r = 0.97) than in diseased human tissue (r = 0.59). Normalization of maximal force to strip cross-sectional area (i.e. to stress) showed considerable more variation (% coefficient of variance) than the normalization to strip actin content in healthy guinea pig tissue, although both strip cross-sectional area and actin content per se showed little variation. Normalization to either parameter did not result in an improved correlation or a decreased variation in the case of diseased human gallbladder tissue.. Normalization of muscle strip force in diseased tissue is questionable, as the assumptions made for healthy tissue are not valid.

Topics: Acetylcholine; Actins; Animals; Cholelithiasis; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Gallbladder; Guinea Pigs; Humans; In Vitro Techniques; Inflammation; Muscle Contraction; Sincalide; Stress, Mechanical

Patients referred to general surgeons for the treatment of gall-bladder stones were studied to evaluate the role of sincalide cholescintigraphy as a gall-bladder stress test in an effort to identify a group of patients whose pain was non-biliary in origin and who would not be improved by cholecystectomy.. Ten asymptomatic controls and 57 patients with gallstones and abdominal symptoms were studied. All patients were interviewed by an independent assessor who identified a group of patients in whom the role of gallstones in their presentation was uncertain (clinically possibly biliary group). All patients and controls underwent sincalide cholescintigraphy. The surgeons remained blinded to the study results throughout the study period. All patients were re-evaluated 6-12 months later to establish the ultimate diagnosis based on their therapeutic response.. Several parameters of gall-bladder function were studied from analysis of the sincalide cholescintigram. Lag time, ejection period, ejection rate and ejection fraction did not differ significantly among controls, patients proven to have non-biliary disease and patients proven to have biliary disease. There were significant differences in mean gall-bladder filling fraction between proven biliary and proven non-biliary groups. However, the group of patients with clinically possibly biliary symptoms could not accurately be separated into those who benefited from cholecystectomy and those who improved without surgery on the basis of this parameter.. Significant differences in gall-bladder filling fraction between symptomatic and asymptomatic gallstone patients were identified suggesting reduced gall-bladder compliance in symptomatic patients. However, the sincalide cholescintigram failed to emerge as a useful gall-bladder stress test. Even in the 1990s, assessment by an experienced surgeon appears to be the most appropriate way to select patients for cholecystectomy.

Topics: Abdominal Pain; Cholelithiasis; Gallbladder; Humans; Radionuclide Imaging; Radiopharmaceuticals; Sincalide; Technetium Tc 99m Lidofenin

Smooth muscle from gallbladders with cholesterol stones exhibits impaired response to cholecystokinin (CCK). This study investigated whether the impaired response is mediated by different signal-transduction pathways responsible for CCK-induced contraction in prairie dog and human gallbladders with cholesterol stones. Gallbladder muscle cells were isolated enzymatically to study contraction. Protein kinase C (PKC) activity was measured by examining the phosphorylation of a specific substrate peptide from myelin basic protein Ac-MBP-(4-14). Gallbladder muscle cells from high-cholesterol-fed prairie dogs contracted less in response to CCK octapeptide (CCK-8) than those from the control group. However, inositol-1,4,5-trisphosphate (IP3), diacylglycerol, and guanosine 5'-O-(3-thiotriphosphate) induced the same magnitudes of contraction in these two groups. In control prairie dog and human gallbladders, the maximal contraction caused by 10(-8) M CCK-8 was blocked by the calmodulin antagonist CGS9343B but not by the PKC inhibitor H-7. Conversely, in gallbladders with cholesterol stones from prairie dogs or human patients, the maximal contraction induced by 10(-8) M CCK-8 was blocked by H-7 and chelerythrine but not by CGS9343B. In these gallbladders CCK-8 caused a significant PKC translocation from the cytosol to the membrane. High CCK concentrations may activate the calmodulin-dependent pathway in functionally normal gallbladder muscle and the PKC-dependent pathway in muscle from gallbladders with cholesterol stones. The defect of gallbladder muscle after cholesterol feeding and stones might reside in the steps before G protein activation.

Topics: 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; Alkaloids; Animals; Benzimidazoles; Benzophenanthridines; Biological Transport; Calmodulin; Cholecystokinin; Cholelithiasis; Enzyme Inhibitors; Gallbladder; Humans; Male; Muscle Contraction; Muscle, Smooth; Phenanthridines; Protein Kinase C; Reference Values; Sciuridae; Sincalide

This study addresses cholecystokinin (CCK)-receptor alterations in stone-diseased and stone-free human gallbladders using different CCK-fragments.. Serosa-free muscle strips were mounted in a modified Krebs-Henseleit-solution of 37 degrees C and aerated with carbogen. The following concentrations of CCK-fragments (CCK 26-33, N-Acetyl CCK 27-33 sulf., CCK 26-29 sulf., CCK 25-33 sulf.) were achieved: 0.1 nmol, 0.5 nmol, 2 nmol, 10 nmol, 100 nmol.. Stone-diseased gallbladders were classified into two groups based on their in vitro reaction to CCK 26-33 (CCK-octapeptide). Muscle strips not contracting below 10 nmol were assigned to the subcontractor group. Histologically scarification, necrosis and signs of severe inflammation of the mucosa were seen in 76.9% of this group. Those starting contractions at 0.1 nmol (like the control group) were called the contractor group. This group had a shallow mucosa and mild inflammatory signs in 54.5%. The sub-contractor group showed higher spontaneous phasic activity at lower tonic activity than the contractor and control groups. In the sub-contractor group CCK 27-33 caused several times higher contractions than all other fragments. A maximal contraction level in the contractor and control groups was reached by CCK 25-33.. This striking effect of CCK 27-33 in the sub-contractor group favors the view of CCK-receptor structural alteration in a subgroup of patients with cholecystolithiasis.

Topics: Adult; Aged; Cholelithiasis; Female; Gallbladder; Humans; In Vitro Techniques; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Peptide Fragments; Receptors, Cholecystokinin; Reference Values; Sincalide

To evaluate the measurement with ultrasonography (US) of the gallbladder ejection fraction after slow-infusion cholecystokinin stimulation in patients with biliary symptoms and in individuals without symptoms.. Gallbladder volumes were calculated in 60 healthy volunteers after a 30-minute infusion of sincalide. The time to maximum response, the gallbladder ejection fraction, and the rate of initial contraction were obtained at US. A total of 100 symptomatic patients were evaluated with this technique. Reference standards included surgical outcome or results of clinical follow-up of at least 1 year.. The average ejection fraction +/- 2 standard deviations was 80% +/- 30. A fraction greater than 60% was considered to be a normal response to cholecystokinin stimulation. There was no statistically significant sex difference. Slow infusion did not produce any side effects. A sensitivity of 75% and a specificity of 100% for determination of gallbladder ejection fraction at US were obtained in patients with surgical and histopathologic proof of disease.. The slow-infusion method is reliable, safe, and reproducible in evaluating gallbladder contraction. The cholecystokinin-stimulated gallbladder ejection fraction test may be useful in determining which patients could benefit from surgery.

Topics: Adult; Aged; Aged, 80 and over; Case-Control Studies; Cholecystitis; Cholelithiasis; Female; Follow-Up Studies; Gallbladder Emptying; Humans; Infusions, Intravenous; Male; Middle Aged; Reproducibility of Results; Sensitivity and Specificity; Sincalide; Ultrasonography

Gastric surgery induces an increased incidence of gallstones. To investigate the changes in gallbladder kinetics after gastric resection, 20 male patients were studied: ten patients undergoing cholecystectomy for gallstones developed after Billroth II gastric resection and ten patients undergoing cholecystectomy for cholelithiasis without previous abdominal surgery. Longitudinal strips from the gallbladder wall were suspended in an organ bath and the isometric tension recorded. Dose-response curves to cholecystokinin-octapeptide and carbachol were obtained. Half the maximal response to cholecysto-kinin-octapeptide was 0.50 +/- 0.11 x 10(-7) M in the first group and 1.36 +/- 0.37 x 10(-7) M in the second group (P < 0.05). The ED50 to carbachol was 24.33 +/- 2.69 x 10(-7) M in the gastrectomy group and 40.39 +/- 5.01 x 10(-7) M in the control group (P < 0.01). There was no significant difference in the maximal contractile response either to cholecystokinin-octa-peptide or carbachol in the two groups. Our study shows an increased gallbladder sensitivity to cholecystokinin-octapeptide and carbachol in patients with gallstones developed after Billroth II gastric resection.

Topics: Aged; Aged, 80 and over; Carbachol; Cholecystectomy; Cholelithiasis; Duodenal Ulcer; Gallbladder; Humans; In Vitro Techniques; Isometric Contraction; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Parasympathomimetics; Sincalide; Stimulation, Chemical; Stomach

Human gallbladders were used to investigate the mechanisms of the impaired contraction induced by cholecystokinin (CCK) associated with cholesterol stones. Single muscle cells were isolated enzymatically with collagenase. Inositol 1,4,5-trisphosphate was measured by high-performance liquid chromatography. Diacylglycerol was assayed by thin-layer chromatography. CCK stimulation showed decreased muscle contraction and production of inositol 1,4,5-trisphosphate and diacylglycerol in gallbladders with cholesterol stones compared with those with pigment stones. Exogenous calmodulin induced maximal contraction of 22.4 +/- 0.5 and 21.0 +/- 0.6% in gallbladders with cholesterol and pigment stones, respectively. Similar findings were observed with a synthetic diacylglycerol analogue. Two G protein activators, aluminum fluoride and guanosine 5'-O-(3-thiotriphosphate), evoked similar responses in these two types of gallbladders, with maximal contractions of 21.3 +/- 0.4 and 23.3 +/- 0.5%, respectively, in those with cholesterol stones and 20.9 +/- 0.8 and 22.6 +/- 0.4%, respectively, in those with pigment stones. These results suggest that receptor-dependent ligands like CCK cannot fully activate the intracellular pathways, which, however, can be fully stimulated by circumventing receptors with G protein activators or second messengers. After G protein activation, the pathways appear to be functionally intact. The defect might then reside in the receptor or in the interaction between receptors and G proteins.

Topics: Aluminum Compounds; Calmodulin; Cholecystokinin; Cholelithiasis; Cholesterol; Diglycerides; Dose-Response Relationship, Drug; Female; Fluorides; Gallbladder; GTP-Binding Proteins; Guanosine 5'-O-(3-Thiotriphosphate); Humans; Inositol 1,4,5-Trisphosphate; Male; Middle Aged; Muscle Contraction; Signal Transduction; Sincalide

It is becoming increasingly evident from a number of studies that endoscopic ultrasound (EUS) is much more sensitive in the diagnosis of cholecystitis than transabdominal ultrasound (TUS). The present study was undertaken to further evaluate this relative sensitivity.. Sixty-six patients with biliary-type pain and a negative transabdominal ultrasound examination underwent combined endoscopic ultrasound and stimulated biliary drainage (EUS/SBD). Stimulated biliary drainage was obtained following intraduodenal infusion of magnesium sulfate or intravenous sincalide, a CCK analogue. EUS was considered positive if sludge or small stones were seen in the gallbladder. Stimulated biliary drainage was considered positive if calcium bilirubinate granules or cholesterol crystals were seen on microscopic examination of aspirated bile.. At operation, 61 of the patients had cholecystitis documented histologically. Fifty-eight of the patients had gallbladder sludge or small stones on EUS. One patient had a negative EUS, but had calcium bilirubinate granules in the bile. Twenty-one patients were followed post-operatively for a period of seven to 17 months, with an average of 10.5 months. Nineteen patients (90.5%) remain free of biliary pain.. Combined endoscopic ultrasound and stimulated biliary drainage (EUS/SBD) had a high sensitivity of 92.4% and a positive predictive value of 100% in the diagnosis of cholecystitis when transabdominal ultrasound was negative. A significant majority (90.5%) of patients with positive EUS/SBD who underwent cholecystectomy had resolution of their biliary pain.

Topics: Cathartics; Cholecystectomy; Cholecystitis; Cholelithiasis; Diagnosis, Differential; Duodenoscopy; Follow-Up Studies; Gallbladder Emptying; Gastrointestinal Agents; Humans; Magnesium Sulfate; Sensitivity and Specificity; Sincalide; Treatment Outcome; Ultrasonography

To examine whether the contractile response of gallbladder smooth muscle cells to cholecystokinin is reduced in patients with gallstones, smooth muscle cells were isolated from the gallbladders of patients either with or without gallstones and their direct contractile responses to cholecystokinin-octapeptide (CCK-8) were examined at physiological concentrations. The basal cell lengths were similar in patients with cholesterol gallstones (43 +/- 1.2 micron (SEM)), in patients with black pigment stones (42 +/- 1.1), and in the gallstone-free group (42 +/- 1.3). The contractile responses to CCK-8 at physiological concentrations ranging from 10(-13) to 10(-11) M were significantly greater in the gallstone patients compared with those in the gallstone-free patients, while the responses were similar at 10(-10) and 10(-9) M. No difference in contractile response to CCK-8 was found between the patients with cholesterol gallstones and those with black pigment stones. The effective dose of 50% was 1.6 x 10(-13) M in cholesterol gallstone patients, 3.2 x 10(-14) M in black pigment stone patients, and 1.0 x 10(-12) M in gallstone-free patients. The current in vitro study showed that the contractile responses of isolated smooth muscle cells of the gallbladder to CCK-8 are not impaired in patients with gallstones.

Topics: Analysis of Variance; Cholecystectomy; Cholelithiasis; Female; Gallbladder; Humans; In Vitro Techniques; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Sincalide; Stomach Neoplasms

This study describes the influence of endogenous and exogenous prostaglandins upon CCK-induced motility patterns of human gallbladders with and without stones (indomethacin and nocloprost; an exogenous PGE2-analogon). From 48 gallbladders with- and 22 gallbladders without stones (control group) longitudinal muscle stripes were dissected and transferred to an organ bath and CCK, indomethacin and nocloprost dose response curves were established. In another experimental protocol, the effect of CCK after indomethacin or nocloprost preincubation is demonstrated. Moreover, specimens of gallbladders were taken for histology and gallstones for analyse. The results demonstrate that gallbladders with stones have a significant higher basic tonus and phasic activities compared to the stone-free controls. Because of these different responses to CCK, gallbladders of the stone-diseased group were divided in two groups: 64% of the gallbladders show a sensitivity and tonic response to CCK like the controls (contractors), 36% demonstrate a reduced sensitivity to CCK and only a slight tonic response (non-contractors). Indomethacin causes a fall in tonus in both stone-diseased groups. It stops spontaneous activity in the contractor and non-contractor group. With indomethacin preincubation all three groups response to CCK with a significant reduced sensitivity. CCK-induced activity is reduced in the control and contractor group. In the non-contractor group, muscle strips do not contract after indomethacin preincubation. Nocloprost induces significant contractions in the control and contractor group. In both groups, the response to CCK after nocloprost preincubation is stronger than the reaction without preincubation. In the non-contractor group, a change in tonus after nocloprost application cannot be demonstrated, there also is no response to CCK after nocloprost preincubation. These results corroborate the notion of a significant contribution of the endogenous prostaglandin system to the regulation of gallbladder motility by CCK.

Topics: Adult; Aged; Aged, 80 and over; Cholecystokinin; Cholelithiasis; Culture Techniques; Dose-Response Relationship, Drug; Female; Gallbladder Emptying; Humans; Indomethacin; Male; Middle Aged; Muscle Contraction; Muscle, Smooth; Prostaglandins; Prostaglandins F, Synthetic; Sincalide; Vasodilator Agents

Gallbladder motility is impaired in specimens with cholesterol stones but normal with pigment stones.. Muscle cells obtained from 19 human gallbladders with cholesterol stones and 11 with pigment stones were enzymatically digested and contracted with cholecystokinin octapeptide (CCK-8), acetylcholine, and KCl.. Muscle cells from pigment stones had a greater contraction than cells from cholesterol stones. CCK-8-induced contraction was unaffected by calcium-free media but was blocked by strontium. Potassium-evoked contraction was blocked by a calcium-free media and unaffected by strontium. Inositol triphosphate (IP-3)-induced contraction was similar to the contraction caused by CCK-8 in permeable cells from pigment stones but was greater than the response to CCK-8 in cells from cholesterol stones.. Muscle cells from gallbladders with cholesterol stones contract less than cells from gallbladders with pigment stones; CCK-8-induced contraction only uses stored calcium; and IP-3 causes contractions of equal magnitude in cells from gallbladders with cholesterol and pigment stones. These abnormalities could result from an impaired receptor activation of the mechanism for IP-3 generation and release of stored calcium.

Topics: Acetylcholine; Adult; Aged; Calcium; Cholelithiasis; Cholesterol; Female; Gallbladder; Humans; In Vitro Techniques; Inositol 1,4,5-Trisphosphate; Male; Middle Aged; Muscle Contraction; Potassium; Sincalide

Thirty patients with chronic upper abdominal pain and no evidence of cholelithiasis were entered into this study. All had negative ultrasonography of the gallbladder, and most had a host of other negative investigations. These patients were referred to a surgeon to evaluate the possibility of atypical biliary colic associated with chronic acalculous cholecystitis. All patients underwent cholecystokinin-stimulated cholescintigraphy and were offered cholecystectomy if the ejection fraction was less than 35 per cent. Of the 30 patients, 27 (90%) had pathologically abnormal gallbladders. Follow-up averaged over 1 year (13.2 mo), and relief of symptoms occurred in 28 (94%). The authors conclude that in appropriately selected patients with symptoms of biliary colic (typical or atypical) and no evidence of cholelithiasis, a cholecystokinin-stimulated cholescintigram is a significant help in predicting not only which patients have gallbladder disease, but also how likely cholecystectomy is to result in an improvement in their symptoms.

Topics: Adolescent; Adult; Cholecystectomy; Cholecystitis; Cholecystography; Cholelithiasis; Chronic Disease; Diagnosis, Differential; Female; Humans; Male; Middle Aged; Pain; Radionuclide Imaging; Sincalide

In light of the effects of gastrointestinal (GI) peptides on bile secretion and biliary tract mobility, we studied the effects of GI peptides on gallstone formation in guinea pigs fed on low protein lithogenic diet. The peptides under study included cholecystokinin octapeptide (CCK-8), vasoactive intestinal peptide (VIP), somatostatin (SRIF), secretin (SEC), and neurotensin (NT). Hepatic bile flow, electrolytes, and other bile components were also measured. It was found that CCK-8 and VIP suppressed the formation of gallstones and increased hepatic bile flow and Na+, K+, Cl- output significantly. On the other hand, SRIF significantly promoted gallstone formation. The rates of gallstone formation in CCK-8, VIP, and SRIF treated guinea pigs were 15.4%, 23.5%, and 88.0%, respectively, in contrast to 56.8% in the control group. The inhibitory effect of CCK-8 and promoting effect of SRIF on gallstone formation were dose-dependent.

Topics: Animals; Bile; Cholelithiasis; Female; Gastrointestinal Hormones; Guinea Pigs; Male; Neurotensin; Sincalide; Somatostatin; Vasoactive Intestinal Peptide

Twenty-five gallbladders were studied in vitro. Sixteen had radiolucent gallstones and 9 had radiopaque gallstones. The radiolucent gallstones had a cholesterol content of 94.17 +/- 3.76% and the radiopaque gallstones had a cholesterol content of 56.6 +/- 4.46%. Half the maximal response (ED50) to cholecystokinin octapeptide (CCK-OP) and to carbachol in strips from patients with radiolucent gallstones was 0.8 +/- 0.15 and 27.01 +/- 3.74 x 10(-7) M, respectively. In strips from patients with radiopaque gallstones, the ED50 was 0.4 +/- 0.08 and 14.92 +/- 3.07 x 10(-7) M, respectively. The ED50 values to CCK-OP and carbachol were greater in strips from specimens with radiolucent gallstones than in strips from specimens with radiopaque gallstones (p less than 0.05). There was no significant difference in the maximal contractile response of the two groups. It can be concluded that gallbladder sensitivity to CCK-OP and carbachol can be modified in relation to differences in the cholesterol and calcium content of the stones.

Topics: Adult; Aged; Carbachol; Cholelithiasis; Gallbladder; Humans; In Vitro Techniques; Male; Middle Aged; Muscle Contraction; Radiography; Sincalide; Ultrasonography

Our aim was to define mechanisms whereby conjugated estrogens (Premarin, exogenous estrogen; Ayerst Laboratories, New York) increase the risk of developing cholesterol gallstones and to determine the role, if any, of dietary cholesterol. We studied gallbladder motor function, biliary lipid composition and secretion, cholesterol absorption, cholesterol synthesis and esterification by peripheral blood mononuclear cells, the clearance of chylomicron remnants, and bile acid kinetics in 29 anovulatory women. 13 were studied on both a low (443 +/- 119 mumol/d) and high (2,021 +/- 262 mumol/d) cholesterol diet. Premarin increased the lithogenic index of bile (P less than 0.05), increased biliary cholesterol secretion (P less than 0.005), lowered chenodeoxycholate (CDCA) pool (P less than 0.001) and synthesis (P less than 0.05), altered biliary bile acid composition [( CA + DCA]/CDCA increases, P less than 0.005), stimulated cholesterol esterification (P less than 0.03), and enhanced the clearance of chylomicron remnants (P = 0.07). Increases in dietary cholesterol stimulated the biliary secretion of cholesterol (P = 0.07), bile acid (P less than 0.05), phospholipid (P = 0.07), and as a result, did not alter lithogenic index. The reduction in CDCA pool and synthesis by Premarin was reversed by increasing dietary cholesterol. Off Premarin, only 24% of the increase in cholesterol entering the body in the diet was recovered as biliary cholesterol or newly synthesized bile acid. On Premarin, 68% of this increase in cholesterol was recovered as these biliary lipids. We conclude that Premarin increases biliary cholesterol by enhancing hepatic lipoprotein uptake and inhibiting bile acid synthesis. These actions of Premarin divert dietary cholesterol into bile.

Topics: Adult; Bile; Bile Acids and Salts; Cholelithiasis; Cholesterol; Cholesterol, Dietary; Chylomicrons; Estrogens, Conjugated (USP); Female; Gallbladder; Humans; Lipid Metabolism; Liver; Middle Aged; Sincalide

Feeding prairie dogs a diet rich in cholesterol induces gallstone formation that is preceded by a sustained decrease in gallbladder smooth muscle contractility. Sphincterotomy is known to prevent gallstone formation in cholesterol-fed prairie dogs. Experiments were designed to determine whether the effect of sphincterotomy is a consequence of hepatic bile diversion, and whether bile diversion prevents the altered contractility. Following sham operation, surgical biliary enteric bypass, or sphincterotomy, prairie dogs were fed a high-cholesterol or a regular diet. Gallbladder muscle contractility and the presence of crystals and stones were determined. In sham-operated animals, the cholesterol diet induced a decrease in gallbladder muscle contractility and caused the formation of cholesterol gallstones. In animals with bile diversion and sphincterotomy, the effects of cholesterol feeding were reduced or prevented. Thus, these procedures may prevent stone formation by preventing a reduction in gallbladder contractility. Contractility was depressed in animals with bile diversion fed a regular diet, compared with animals with a sham operation fed a regular diet. The mechanism for this depression may differ from that induced by the cholesterol diet. Diversion, and perhaps sphincterotomy, impairs gallbladder filling. Thus, gallbladder muscle is not stretched and does not contract against a load. This could result in a "disuse atrophy." If the results from our study apply to humans, sphincterotomy may reduce stone formation by preventing the effects of lithogenic bile on gallbladder muscle contractility and by enhancing the ability of the muscle to empty the lithogenic bile.

Topics: Animals; Bile; Catheterization; Cholelithiasis; Cholesterol, Dietary; Common Bile Duct; Duodenum; Gallbladder; In Vitro Techniques; Ligation; Male; Muscle Contraction; Sciuridae; Sincalide; Sphincter of Oddi

It has been postulated that cholecystokinin sonography may be useful in the diagnosis of acute acalculous cholecystitis in the hospitalized patient. To evaluate this hypothesis, sincalide, a cholecystokinin derivative, was administered to 15 fasting trauma patients who had undergone laparotomy. No biliary or gallbladder disease was found in any patient. Sincalide was slowly administered intravenously, and the gallbladder was examined with ultrasound every 5 minutes for 60 minutes. The average decreases in length, height, and width of the gallbladder were 15%, 23%, and 21%, respectively. In only four of the 15 patients was there a decrease by more than 50% in any of these dimensions. The average decrease in gallbladder volume was 33% (range, 0%-97%), with no change in gallbladder volume in four patients. There is considerable variability in gallbladder response to administration of sincalide in the fasting hospitalized patient. Lack of contraction of the gallbladder after injection of cholecystokinin should not be considered a major criterion in the diagnosis of acute acalculous cholecystitis.

Topics: Acute Disease; Adolescent; Adult; Cholecystitis; Cholelithiasis; Female; Gallbladder; Humans; Male; Middle Aged; Sincalide; Ultrasonography

The pathogenesis of cholesterol cholelithiasis in man is probably multifactorial and the mechanism by which gallbladder stasis occurs in gallstone patients has not been studied in detail. In the present study, time-dependent changes in gallbladder motility of the Richardson Ground Squirrels were investigated. 100 of animals were examined for their contractile responses of gallbladders to CCK-OP and Ach in vitro. Also, biochemical changes of serum and bile were investigated. There were no significant differences in motility of gallbladders to CCK-OP or Ach between control and cholesterol-fed animals. The results of this study indicate that gallbladder muscle contractility remains unchanged during the dietary induction of cholesterol gallstones in the Richardson Ground Squirrel.

Topics: Acetylcholine; Animals; Bile; Cholelithiasis; Cholesterol; Female; Gallbladder; In Vitro Techniques; Male; Movement; Muscle Contraction; Muscle, Smooth; Sciuridae; Sincalide; Time Factors

The purpose of this study was to evaluate the effect of age and the role of cholecystokinin therapy on gallstone formation in guinea pigs. Guinea pigs (31 1-mo-old, 31 1-yr-old, and 23 3-yr-old) were placed on a cholelithogenic diet for 2 wk while another 10 guinea pigs of each age group remained on regular chow. Half of each group received a daily injection of cholecystokinin (0.5 nmol/kg). After 2 wk, guinea pigs were killed and the gallbladders were examined for gallstones. The concentrations of bile constituents were determined. The prevalence of gallstones was: 1-mo-old, control 0 out of 16, cholecystokinin 1 out of 15; 1-yr-old, control 3 out of 14, cholecystokinin 5 out of 16; 3-yr-old, control 10 out of 11, cholecystokinin 3 out of 8. Gallstone formation was significantly greater in 3-yr-old controls than in the two younger control groups, and cholecystokinin treatment significantly reduced the incidence of gallstones to near the level seen in younger guinea pigs. In the two younger age groups (but not in the 3-yr-old group), the cholelithogenic diet significantly reduced the concentration of bile salts in bile below that of guinea pigs on a normal diet. The cholelithogenic diet and treatment with cholecystokinin did not alter the relative compositions of bile lipids from that of guinea pigs on a normal diet in any of the three ages studied. In the second experiment we measured gallbladder emptying in response to exogenous infusion of cholecystokinin-8 (100 fmol/kg/h-100 nmol/kg/h) in the same three age groups of guinea pigs in vivo that had been maintained on regular chow. There was no difference in cholecystokinin sensitivity between the two younger age groups, but both were significantly more sensitive to cholecystokinin than the 3-yr-old guinea pigs in rate of gallbladder emptying in the dose range 1 pmol/kg/h-1 nmol/kg/h. We conclude that a major factor in the increased incidence of gallstone formation in the aged guinea pig gallstone model is decreased gallbladder emptying due to decreased gallbladder sensitivity to cholecystokinin.

Topics: Aging; Animals; Bile; Bile Acids and Salts; Cholelithiasis; Cholesterol, Dietary; Gallbladder; Guinea Pigs; Male; Sincalide

It is well known that stasis of lithogenic bile in the gallbladder is an important factor in cholesterol gallstone formation. In this study, hamsters fed with standard lithogenic diet were given physiologic dose of exogenous cholecystokinin-octapeptide daily to facilitate emptying of the gallbladder. It was found that there was significant reduction in the gallstone formation. This study suggests that gallbladder motility is closely correlated with cholesterol gallstone formation, and administration of exogenous cholecystokinin-octapeptide can effectively prevent gallbladder stasis and reduce the incidence of cholelithiasis. This method may be useful for gallstone prophylaxis in high-risk individuals.

Topics: Animals; Cholelithiasis; Cholesterol; Cricetinae; Female; Male; Mesocricetus; Sincalide

Stasis of bile within the gallbladder has long been suspected of having an important role in the pathogenesis of gallstone disease. We postulated that the female preponderance of gallstone disease might partly be related to the effects of progesterone, a known smooth muscle relaxant, on specific receptors in the gallbladder wall, leading to stasis of bile. A total of 42 patients with gallstone disease and 28 control subjects underwent radionuclide scan imaging and their gallbladder ejection fractions were calculated in response to intravenous infusion of cholecystokinin octapeptide. Patients then underwent cholecystectomy and a piece of gallbladder fundus was assayed for the presence of progesterone receptors. Receptors were present in 60 percent of patients. As a group, gallstone patients had a decreased ejection fraction compared with control subjects. The presence of progesterone receptors in the gallbladder wall was associated with a decreased percentage of ejection compared with both healthy control subjects and patients whose gallbladders were receptor-negative. We conclude that progesterone receptors are present in the gallbladder wall of gallstone patients and that their presence correlates with impaired gallbladder emptying.

Topics: Adult; Aged; Aged, 80 and over; Cholelithiasis; Female; Gallbladder; Humans; Male; Middle Aged; Muscle Contraction; Radionuclide Imaging; Receptors, Progesterone; Sincalide

Age, gender and female sex hormones are thought to influence contractility of the gallbladder. Surgically removed gallbladders with gallstones and mild chronic cholecystitis from three groups of patients (males and premenopausal and postmenopausal females) were studied. Strips were incubated in Krebs solution, in Krebs containing 0.2 per cent ethanol-propylene glycol solvent (solvent controls) or in Krebs solution containing progesterone (0.1, 1.0 and 10.0 micromolar) or 17-beta-estradiol (0.1 and 1.0 micromolar). Concentration response curves were obtained for acetylcholine and octapeptide of cholecystokinin and the maximum contraction and median effective dose values were recorded. Steroid receptor levels in the wall of the gallbladder were also estimated. In gallbladders from males, the sensitivity to acetylcholine increased significantly with age. However, in gallbladders from premenopausal women, there was a significant decrease in sensitivity to acetylcholine and cholecystokinin with age. Progesterone (1 and 10 micromolar) reduced the spontaneous rhythmic contractions of the strips by 35 +/- 8 and 65 +/- 8 per cent, respectively, and also reduced the maximum contraction of gallbladders from postmenopausal females to acetylcholine. Estradiol-17 beta had no effect upon the contractility of the gallbladder. The results of additional studies showed that the human gallbladder is estrogen and progesterone receptor negative. The sensitivity of the gallbladder to neurohormonal stimulation varies with age and gender. These results indicate that short term exposure to female sex hormones has an inhibitory effect upon gallbladder contractility. The inhibitory effects of short term exposure to progesterone seen in this study are probably nonspecific in nature.

Topics: Acetylcholine; Adult; Aging; Cholelithiasis; Estradiol; Female; Gallbladder; Humans; In Vitro Techniques; Male; Menopause; Muscle Contraction; Progesterone; Sex Factors; Sincalide

We studied gallbladder bile flow before, during, and after cholesterol gallstone formation in the prairie dog using infusion cholescintigraphy with 99mTc-diethyl iminodiacetic acid. In 18 fasting animals partitioning of bile between gallbladder and intestine was determined every 15 min for 140 min, and gallbladder response to cholecystokinin (5 U/kg X h) was calculated from the gallbladder ejection fraction. Ten prairie dogs were then placed on a 0.4% cholesterol diet and 8 on a regular diet, and the studies were repeated 1, 2, and 6 wk later. The proportion of hepatic bile that entered the gallbladder relative to the intestine varied from one 15-min period to the next, and averaged 28.2% +/- 5.1% at 140 min. Partial spontaneous gallbladder emptying (ejection fraction 11.5% +/- 5.6%) was intermittently observed. Neither the number nor the ejection fraction of spontaneous gallbladder contractions changed during gallstone formation. By contrast, the percent of gallbladder emptying in response to cholecystokinin decreased from 72.1% +/- 5% to 25.9% +/- 9.3% (p less than 0.025) in the first week and was 14.3% +/- 5.5% at 6 wk (p less than 0.01 from prediet values, not significant from first week). Gallbladder filling decreased from 28.2% +/- 5.1% to 6.7% +/- 3% (p less than 0.01), but this change was only observed after 6 wk, when gallstones had formed. This study shows that bile flow into the gallbladder during fasting is not constant; the gallbladder contracts intermittently; gallbladder emptying in response to exogenous cholecystokinin is altered very early during gallstone formation; and gallbladder filling remains unaffected until later stages, when gallstones have formed.

Topics: Animals; Bile; Cholecystokinin; Cholelithiasis; Cholesterol; Cholesterol, Dietary; Gallbladder; Imino Acids; Male; Muscle Contraction; Radionuclide Imaging; Sciuridae; Sincalide; Technetium; Technetium Tc 99m Diethyl-iminodiacetic Acid; Time Factors

Some investigators have reported that patients with gallstones empty their gallbladders more rapidly than do healthy subjects. This may contribute to the formation of lithogenic bile. To date, cholecystokinin is considered the prime mediator of gallbladder contraction. Evidence exists that cholecystokinin may not be the major hormone accounting for gallbladder emptying. The purpose of this study was to demonstrate the existence of this noncholecystokinin substance in healthy persons and to compare its concentration with that in patients with cholesterol gallstones. Fasting serum levels from 15 healthy human subjects (8 women and 7 men, mean age 32 +/- 8 years) and 10 patients with cholesterol gallstones (5 women and 5 men, mean age 48 +/- 16 years) were studied. Using rabbit in vitro gallbladder bioassay and cholecystokinin-8 as standards, serum bioactivity was measured and expressed as cholecystokinin-8 equivalent bioactivity. The effectiveness of serum to contract the gallbladder was tested before and after removal of cholecystokinin from the serum. Cholecystokinin was removed from the serum samples by affinity chromatography with Sepharose 4B beads coated with cholecystokinin 5135 antibody. Gallbladder contractility from this treated serum thus reflects the action of a noncholecystokinin stimulant. The cholecystokinin-8 bioactivity equivalents in untreated samples from healthy subjects and from patients with gallstones were 2.9 +/- 0.3 and 7.6 +/- 0.7 ng/ml, respectively. The fact that bioactivity of serum persisted after removal of cholecystokinin in both groups of subjects provides evidence that a noncholecystokinin stimulant of gallbladder contraction exists. This substance is found in significantly higher concentrations in the fasting serum of patients with gallstones compared with healthy subjects. This finding may explain, at least in part, the increased gallbladder emptying rate in patients with gallstones and may account for the reduced bile salt pool size and, thus, formation of lithogenic bile.

Topics: Adult; Animals; Biological Assay; Cholelithiasis; Female; Gallbladder; Humans; In Vitro Techniques; Male; Middle Aged; Rabbits; Sincalide

Altered gallbladder motility could predispose to, or result from, gallstone formation and could also explain the alleged relief of biliary colic seen during bile acid therapy. Therefore, in 14 controls, 25 patients with radiolucent gallstones, and 14 patients with radiopaque gallstones, we used two techniques to measure gallbladder contraction--radionuclide imaging and real-time ultrasound--in response to one of two stimuli--a Lundh meal or intravenous cholecystokinin-octapeptide. Using the radionuclide technique, postprandial gallbladder emptying (t1/2) was prolonged (p less than 0.01) both in patients with radiopaque (26.7 +/- 3.1 min, mean +/- SEM) and radiolucent (21.7 +/- 3.1) gallstones when compared with controls (10.2 +/- 1.5). In patients with radiolucent stones, the t1/2 of gallbladder emptying became further prolonged (p less than 0.05) after 1 mo of therapy with 8-10 mg/kg body wt X day of ursodeoxycholic acid, to 32.1 +/- 4.4 min. A similar pattern of results was seen after cholecystokinin-octapeptide and also with real-time ultrasound. Thus, after both stimuli and using two independent techniques, gallbladder contraction was reduced in patients with gallstones. The slower and less complete gallbladder emptying with ursotherapy might explain the reduction in biliary colic noted during treatment.

Topics: Adult; Aged; Cholelithiasis; Deoxycholic Acid; Eating; Female; Gallbladder; Humans; Imino Acids; Infusions, Parenteral; Male; Middle Aged; Muscle Contraction; Radionuclide Imaging; Sincalide; Technetium; Technetium Tc 99m Lidofenin; Time Factors; Ultrasonography; Ursodeoxycholic Acid

Several human and experimental observations suggest that gallbladder stasis is an important link between the hepatic secretion of cholesterol saturated bile and the formation of cholesterol gallstones. In the cholesterol-fed prairie dog model, gallbladder stasis occurs before gallstone formation. In this study we sought to determine the specific defects in extrahepatic biliary physiology responsible for gallbladder stasis in this model. Adult male prairie dogs were fed either a trace cholesterol or a 0.4% cholesterol-enriched diet. In acute terminal experiments, gallbladder contents were examined for cholesterol crystals and gallstones, and gallbladder function was determined at rest and in response to intravenous cholecystokinin-octapeptide. The following alterations in gallbladder function developed concurrently with biliary cholesterol crystallization, but before gallstone formation: (a) decreased gallbladder emptying, (b) increased intragallbladder pressure in response to cholecystokinin-octapeptide, (c) increased cystic duct closing pressure, and (d) increased resistance to outflow through the cystic duct.

Topics: Animals; Cholecystokinin; Cholelithiasis; Cholestasis, Extrahepatic; Cholesterol, Dietary; Cystic Duct; Gallbladder; Male; Peptide Fragments; Pressure; Rodentia; Sincalide; Stimulation, Chemical

To examine the effect of changes in bile lithogenicity on gallbladder muscle function, in vitro gallbladder contractility was studied in an animal model of cholesterol gallstones: Richardson ground squirrels fed either a trace (control) or a 1% wt/wt cholesterol (test) diet. Lithogenic index of gallbladder bile increased on the test diet from 0.52 +/- 0.03 to 0.81 +/- 0.04 (p less than 0.001). Isometric tensions generated in response to cholecystokinin-octapeptide, acetylcholine, or potassium depolarization, were all reduced greater than 50% in the test gallbladder muscles (p less than 0.05), without any significant shift of the normalized dose-response curves. Tension in response to cholecystokinin-octapeptide differed significantly (p less than 0.05) between each stage of stone formation compared with controls: 42% decrease in test animals before development of stones; 65% decrease in those with gallstones. Ileal muscle from these animals, when tested with the same three stimuli, showed no adverse effects of the high-cholesterol diet. Another animal model, the prairie dog, also demonstrated a similar in vitro defect in gallbladder contractility associated with increases in bile lithogenicity. Thus, in the ground squirrel, a progressive defect in smooth muscle contractility to three different stimuli coincides with early changes in bile lithogenicity. The defect is not associated with any loss of sensitivity to these stimuli, and appears to be localized specifically to the gallbladder muscle. Its presence in two animal models of cholelithiasis suggests that biliary stasis is an important factor in the early stages of cholesterol stone formation.

Topics: Acetylcholine; Animals; Cholecystokinin; Cholelithiasis; Cholesterol; Cholesterol, Dietary; Gallbladder; In Vitro Techniques; Intestines; Male; Muscle Contraction; Muscle, Smooth; Peptide Fragments; Potassium; Sciuridae; Sincalide

Topics: Administration, Intranasal; Animals; Cholecystography; Cholecystokinin; Cholelithiasis; Dogs; Female; Injections, Intravenous; Injections, Subcutaneous; Male; Rectum; Sincalide; Suppositories