Specific somatostatin-binding to cytosol of bovine gallbladder mucosa

The binding of somatostatin was studied in the cytosolic fraction of bovine gallbladder mucosa. The binding reaction depended on time, temperature and pH, and was reversible, saturable and specific. Stoichiometric data suggested the presence of two classes of binding sites: a class with high affinity (K _d=23.6 nM) and low capacity (3.7 pmol somatostatin/mg protein) and a class with low affinity (K _d=284.6 nM) and high capacity (85.0 pmol somatostatin/mg protein) at 37°C and pH 7.4. The binding sites were highly specific for somatostatin since peptides such as [Leu]enkephalin, neurotensin, vasoactive intestinal peptide and substance P showed practically no effect upon somatostatin binding. The presence of somatostatin-binding sites in the cytosolic fraction of gallbladder mucosa, together with the known occurrence of somatostatin nerve endings in the gallbladder strongly suggests that this peptide may be involved in the physiology and physiopathology of gallbladder mucosa.     

清胆颗粒利胆作用的实验研究

目的：通过研究清胆颗粒对正常大鼠及模型动物胆汁及病理组织的影响,确定其利胆功效,为临床应用提供依据.方法：采用胆管引流法测定清胆颗粒对正常大鼠胆汁量的影响;应用石胆酸（lithocholic acid,LCA）造成豚鼠胆囊炎模型,测定其对模型动物肝重、肝指数、胆囊容积及胆囊病理组织的影响.结果：清胆颗粒能显著增加正常大鼠胆汁流量,明显减轻模型动物肝湿重和肝指数,缩小胆囊容积,能明显降低胆汁中TB、DB及UCB/TB含量,对Ca^2＋增高有明显的抑制作用,对模型动物胆囊粘膜上皮增生有明显的抑制作用.结论：清胆颗粒具有显著的利胆作用.     

Beta-endorphin in the human gallbladder

The presence of beta-endorphin-like immunoreactivity (beta-END-LI) in human gallbladder and its release were examined by means of radioimmunochemical measurements and immunohistochemical stainings. beta-END-LI was detected in the gallbladder (27.2 +/- 3.2 ng/g wet weight, mean +/- S.E.). The immunoreactivity in beta-END-LI extracted from the gallbladder was similar to that of synthetic beta-END, judging from the result of its inhibition curve parallel to that of the synthetic substance in the radioimmunoassay (RIA) system. On gel-filtration chromatography of a gallbladder extract, two components of beta-END-LI were found; one eluted on a position of beta-lipotropin (beta-LPH) and another on a position of synthetic beta-END. Specific beta-END-LI positive cells were detectable in metaplastic mucous glands. When human gallbladder mucosa was perfused with a solution of 10(-8) M or 10(-6) M cholecystokinin octapeptide (CCK-8), the release of beta-END-LI from mucosa into the perfusate increased 2-3 fold. These results indicate that beta-END-LI present in human gallbladder is released by the direct action of CCK-8 on the gallbladder mucosa and suggest that it may have a physiological or pathophysiological role.     

丹顶鹤胆囊的显微观察

应用光镜及透射电镜观察一例成体雄性丹顶鹤胆囊。结果表明,丹顶鹤胆囊粘粘膜、肌层和外膜三部分组成。粘膜上皮高柱状、游离端有密集排列的微绒毛,胞质面端有许多粘液颗粒,说明上皮细胞具有吸收功能并可分泌粘液。     

Scanning electron microscopic study of human gallbladder mucosa in cholelithiasis

The mucosa of 17 gallbladders removed because of cholelithiasis was examined by means of transmission and scanning electron microscopy. Scanning electron micrographs revealed a mulberry-like organization of the normal human gallbladder mucosa. In acute inflammation, circumscribed ulcers were often seen; along their margins the epithelial cells were swollen and isolated from the surrounding cells. In chronic cholecystitis there was an extensive lack of epithelium on the irregularly oriented folds of the mucosa and the characteristic mulberry-like organization was preserved only in islets.     

Use of antibiotics and nitrofurans in treating acute and chronic cholecystitis

Sixty-two patients with acute cholecystitis and 108 patients with chronic calculous cholecystitis were examined. High levels of contamination of the bile, gallbladder mucosa and gallstones were shown. E. coli, Staphylococcus and Streptococcus were most frequent among 20 species of aerobic and anaerobic bacteria. Preoperative sanation of the hepatoduodenal area with antibiotics did not result in complete elimination of the bacteria in the bile, gallbladder mucosa and gallstones. The use of nitrofurans and especially furazolidone and furagin in the preoperative period prevented the microbial growth in the specimens collected during the operations. The data of the study allow recommending the use of furazolidone and furagin for preoperative sanation of the biliferous tract.     

Carbamyl phosphate and glutamine stimulation of the gallbladder salt pump

Summary Carbamyl phosphate stimulates neutral salt transport in thein vitro gallbladder. Acetyl phosphate also stimulates transport. In addition, the metabolic precursors of carbamyl phosphate, glutamine and bicarbonate ion, stimulate transport of fluid across the gallbladder mucosa. These data suggest that the effect of glutamine in stimulating increases in transport is achieved via the synthesis of carbamyl phosphate which in turn reacts with the transport mechanism. Metabolic precursors of glutamine, glutamate plus a nitrogen source, when combined will produce an effect similar to glutamine alone. Fluid transport is also slowed by azaserine; thus, the stimulation effect appears to be restricted to glutamine.     

A Comparative Study of Clinicopathological Features between Chronic Cholecystitis Patients with and without Helicobacter pylori Infection in Gallbladder Mucosa

Background

Helicobacter pylori has been isolated from 10%–20% of human chronic cholecystitis specimens but the characteristics of “Helicobacter pylori positive cholecystitis” remains unclear. This study aims to compare the clinicopathological features between chronic cholecystitis patients with and without Helicobacter pylori infection in gallbladder mucosa.

Methods

Three hundred and twenty-six chronic cholecystitis patients were divided into two groups according to whether Helicobacter pylori could be detected by culture, staining or PCR for Helicobacter 16s rRNA gene in gallbladder mucosa. Positive samples were sequenced for Helicobacter pylori-specific identification. Clinical parameters as well as pathological characteristics including some premalignant lesions and the expression levels of iNOS and ROS in gallbladder were compared between the two groups.

Results

Helicobacter pylori infection in gallbladder mucosa was detected in 20.55% of cholecystitis patients. These patients had a higher prevalence of acid regurgitation symptoms (p = 0.001), more histories of chronic gastritis (p = 0.005), gastric ulcer (p = 0.042), duodenal ulcer (p = 0.026) and higher presence of Helicobacter pylori in the stomach as compared to patients without Helicobacter pylori infection in the gallbladder mucosa. Helicobacter pylori 16s rRNA in gallbladder and gastric-duodenal mucosa from the same individual patient had identical sequences. Also, higher incidences of adenomyomatosis (p = 0.012), metaplasia (p = 0.022) and higher enhanced expressions of iNOS and ROS were detected in Helicobacter pylori infected gallbladder mucosa (p<0.05).

Conclusions

Helicobacter pylori infection in gallbladder mucosa is strongly associated with Helicobacter pylori existed in stomach. Helicobacter pylori is also correlated with gallbladder premalignant lesions including metaplasia and adenomyomatosis. The potential mechanism might be related with higher ROS/RNS production but needs further investigation.     

Calcitonin gene-related peptide: an inhibitor of guinea pig gallbladder contraction.

Calcitonin gene-related peptide (CGRP) relaxes vascular and intestinal smooth muscle. This study localized CGRP in the guinea pig gallbladder, examined the effects of CGRP on KCl- and ACh-induced contraction, and determined CGRPs site of action in the gallbladder. The gallbladder of male Hartley guinea pigs was used in in vitro tension studies, radioimmunoassay, or immunocytochemical studies. Radioimmunoassay showed that 8.0 +/- 0.5 pmol/g of immunoreactive CGRP was present. Immunocytochemistry demonstrated that immunoreactive-CGRP nerve fibers occurred around blood vessels, in gallbladder smooth muscle layers, and were associated with ganglia. No immunoreactive cell bodies were observed, even after colchicine treatment. The in vitro tension studies showed that CGRP inhibits either KCl- or acetylcholine-stimulated contraction. CGRP may in part act directly on the gallbladder smooth muscle to inhibit contraction.     

Apoptosis and regeneration of the gastric mucosa under the action of S-phase-specific agent, hydroxyurea

The treatment of periodic injections of hydroxyurea to mice on the processes of regeneration in gastric mucosa was studied. In all experimental groups of animals dystrophic and atrophic changes of gastric mucosa could be observed. The phenomena of dystrophy were more pronounced. If the time of the experiment was not more than 4 days apoptosis bodies could be found only in the epithelium. The long-term duration of the experiment resulted in increased death of apoptosis bodies inside the glands, and development of atrophy of gastric mucosa.     

Polysaccharides and mucin 5AC (MUC5AC) expression in gallbladder mucosa of young patients with gallstones as evaluated by spatial visualization and quantification

The study aimed at examination of tissue expression of polysaccharides and secretory mucin 5AC (MUC5AC) in young patients (up to 25 years of age) with a symptomatic gallstones. For comparison, patients most frequently subjected to cholecystectomy were studied, i.e. patients of approximately 50 years of age with the same diagnosis. In quantitative studies on tissue expression of both mucus components, the modern technique of spatial visualization was applied for the first time. Application of the technique permitted to demonstrate significant positive relationships between expression of glycoproteins (immunocytochemical ABC technique for detection of MUC5AC) and expression of sugar components in mucus (PAS technique) and to confirm suitability of the technique for quantitative appraisal of both histochemical and immunocytochemical reactions. An even higher expression of polysaccharides in the entire mucosa and of MUC5AC was detected in gallbladder epithelium of 50-year-old patients, as compared to young patients with symptomatic gallstones. In the young patients, expression of polysaccharides correlated with inflammatory activity (grading), width of gallbladder wall and PLT level in peripheral blood. A significantly higher expression of polysaccharides in gallbladder epithelium was demonstrated in young patients admitted in the emergency mode to the hospital. These correlations in young patients may suggest a role of both mucus components in pathogenesis of cholelithiasis in this age group. A quantitative appraisal of mucus component expression in the two parts of gallbladder mucosa (epithelium vs. entire mucosa) using spatial visualization technique permitted to more accurately compare production of glycoproteins and of polysaccharides in patients with cholelithiasis and to demonstrate additional correlations of a potential clinical significance.     

Sensory innervation of the ear drum and middle-ear mucosa: Retrograde tracing and immunocytochemistry

Summary The distribution and origin of nerve fibers of presumed sensory nature in the ear drum and middle-ear mucosa of the rat were studied by a retrograde tracing technique in combination with immunocytochemistry.Application of True Blue (TB) on the ear drum or on the middle-ear mucosa labeled nerve cell bodies in the jugular, trigeminal, geniculate and cervical dorsal root ganglia (C₂–C₄). Judging from the number of TB-labeled nerve cell bodies the jugular and trigeminal ganglia contributed the major component to the sensory innervation of the ear drum and the middle-ear mucosa, while the contribution from the geniculate and cervical dorsal root ganglia was relatively minor.The majority of the TB-labeled nerve cell bodies contained calcitonin gene-related peptide (CGRP), whereas minor populations stored substance P (SP) and neurokinin A (NKA). Nerve fibers containing SP, NKA and CGRP were moderate in number in the middle-ear mucosa and few in the ear drum. Double immunostaining revealed that SP invariably coexisted with NKA in nerve cell bodies in the ganglia examined. The SP/NKA-containing nerve cell bodies constituted a subpopulation of those storing CGRP.The findings indicate that several ganglia project to the ear drum and middle-ear mucosa and that many neuropeptides are involved in the mediation of middle-ear sensitivity.     

The effects of mucus on the binding of cationized ferritin by human and animal gastrointestinal epithelium

Human gallbladder and gastric epithelial cells are normally covered with a layer of mucus. When specimens were exposed to cationized ferritin (CF) in vitro, they did not regularly bind nor internalise it. If the tissues were first exposed to the mucolytic agents cysteamine or pepsin, then the gallbladder epithelium readily bound CF and the gastric epithelium irregularly. The in vivo binding of CF by guinea pig gallbladder could be abolished by the induction of mucous hypersecretion by the antibiotic lincomycin. The removal of the mucus by mucolytic agents restored the binding of CF. The irregular binding of CF by gastric mucosa after the use of mucolytic agents suggests other factors may be at play.     

Light and electron microscopic observations of the autonomic innervation of the mouse gallbladder mucosa

Summary The autonomic innervation of the mouse gallbladder mucosa was studied using histo-and cytochemical methods. In a light microscopic investigation the distribution of acetylcholinesterase (AChE) activity and formaldehyde-induced fluorescence was studied histochemically. Nerve fibres and small varicosities showed concentrations of AChE activity very close to the epithelium in the subepithelial connective tissue. No adrenergic nerves were observed in the mucosa.When using the electron microscope and employing the potassium permanganate fixation/staining technique only one sort of axonal enlargement was encountered, viz. the cholinergic type. These varicosities contained numerous agranular vesicles (500–600 Å in diameter). No varicosities of the adrenergic (dense-cored vesicles) type were observed.Signs of increased secretory activity in the epithelium were observed in the first few minutes after cholinergic stimulation. After repeated in vivo stimulation, there was an almost total depletion of glycoprotein granules, best seen when using the cytochemical PA-CrA-silver technique. The findings suggest that the subepithelial connective tissue and the epithelium of the mouse gallbladder mucosa have a cholinergic innervation.     

Ketone body utilization in duodenum. Differential effect of fasting on lipogenesis from acetoacetate and 3-hydroxybutyrate

The effect of fasting and refeeding on oxidation, lipogenesis and amino acid synthesis from ketone bodies has been studied in neonatal chick duodenal mucosa. Oxidation and amino acid synthesis were higher from acetoacetate and were stimulated by fasting from both 3-hydroxybutyrate and acetoacetate. On the contrary, lipogenesis was always higher from 3-hydroxybutyrate and fasting reduced lipogenesis rate from acetoacetate (by 66%) but not from 3-hydroxybutyrate. Results suggests the existence of a cytosolic fast-dependent acetoacetyl-CoA synthetase in chick duodenal mucosa which is involved in phospholipid synthesis.     

Light and electron microscopic observations of the autonomic innervation of the mouse gallbladder mucosa. A histochemical, cytochemical, and secretory study.

The autonomic innervation of the mouse gallbladder mucosa was studied using histo- and cytochemical methods. In a light microscopic investigation the distribution of acetylcholinesterase (AChE) activity and formaldehyde-induced fluorescence was studied histochemically. Nerve fibres and small varicosities showed concentrations of AChE activity very close to the epithelium in the subepithelial connective tissue. No adrenergic nerves were observed in the mucosa. When using the electron microscope and employing the potassium permanganate fixation/staining technique only one sort of axonal enlargement was encountered, viz. the cholinergic type. These varicosities contained numerous agranular vesicles (500-600 A in diameter). No varicosities of the adrenergic (dense-cored vesicles) type were observed. Signs of increased secretory activity in the epithelium were observed in the first few minutes after cholinergic stimulation. After repeated in vivo stimulation, there was an almost total depletion of glycoprotein granules, best seen when using the cytochemical PA-CrA-silver technique. The findings suggest that the subepithelial connective tissue and the epithelium of the mouse gallbladder mucosa have a cholinergic innervation.     

The effects of mucus on the binding of cationized ferritin by human and animal gastrointestinal epithelium

Summary Human gallbladder and gastric epithelial cells are normally covered with a layer of mucus. When specimens were exposed to cationized ferritin (CF) in vitro, they did not regularly bind nor internalise it. If the tissues were first exposed to the mucolytic agents cysteamine or pepsin, then the gallbladder epithelium readily bound CF and the gastric epithelium irregularly. The in vivo binding of CF by guinea pig gallbladder could be abolished by the induction of mucous hypersecretion by the antibiotic lincomycin. The removal of the mucus by mucolytic agents restored the binding of CF. The irregular binding of CF by gastric mucosa after the use of mucolytic agents suggests other factors may be at play.     

Extracellular space determination in gallbladder mucosa

Sucrose seems to be the best marker of the extracellular space in gallbladder mucosa. Inulin space is always smaller than sucrose space and the difference seems to be due to a sweeping-away effect in the intercellular channels as metabolic inhibitors and ouabain reduce it. Mannitol space continuously increases and this substance is likely to enter the cells.     

Association of immune cells with neuroepithelial bodies in the lungs of neonatal dogs,cats and hamsters

A close topographical association between neuroepithelial bodies and immune cells is occasionally observed in the lungs of various neonatal mammalian species. The immune cells concerned are mast cells and neutrophil or eosinophil granulocytes. In the lungs of newborn puppies having undergone left lung autotransplantation, mast cells are particularly numerous in the airway mucosa of both right and left lungs and their association with neuroepithelial bodies is highly significant. Several of the substances known to be synthesized by the neuroepithelial bodies have a chemoattractive effect on immune cells. Thus, our observations indicate that intrapulmonary neuroepithelial bodies contribute to the local immune response.This investigation was supported by a grant from the Fonds voor Geneeskundig Wetenschappelijk Onderzoek (FGWO-NFWO), Belgium     

Nucleus raphe obscurus participates in regulation of gallbladder motility through vagus and sympathetic nerves in rabbits

The aim of the present study is to investigate if the nucleus raphe obscurus (NRO) participate in regulating the gallbladder motility in rabbits. Rabbits were fasted for about 20-24 hours. After anesthetization with urethane, an incision was made at the middle of the abdomen and the gallbladder was exposed. A frog bladder connected with force transducer was inserted into the gallbladder through a small incision at the funds to record gallbladder motility (tonic contraction and phasic contraction). Glutamate and other chemicals were microinjected into NRO through a vitreous tube attached to a microsyringe. We found both the tonic contraction and phasic contraction of the gallbladder were enhanced after the glutamate was injected into NRO. GABA inhibited gallbladder motility if administrated in the same way. Microinjection of ketamine, NMDA (N-methyl-D-aspartate) receptor antagonist, into NRO inhibited the phasic contraction of gallbladder. Administration of CNQX (6-cyano-7-nitroquinoxaline-2, 3-dione), a non-NMDA receptor antagonist, enhanced the gallbladder tonic contraction. Pretreatment of ketamine into NRO attenuated the effect of glutamate, while pretreatment of CNQX had no effect on it. Intravenous injection of atropine or vagotomy completely abolished the effect of glutamate on gallbladder phasic contraction, while intravenous injection of phentolamine or transecting the spinal cord at T3-4 inhibited that on tonic contraction. Intravenous injection of propranolol did not influence the glutamate effect. These results suggested that glutamate in NRO participates in regulating the motility of the gallbladder through NMDA receptor. When excited, the NMDA receptors in NRO enhance the phasic contraction of the gallbladder through vagus nerve and peripheral M-receptors, and enhance the tonic contraction of the gallbladder through sympathetic nerve and peripheral alpha-receptors. GABA in NRO is also involved in the regulation of gallbladder motility.