bluuebunny/crossref_metadata_2025 huggingface dataset:
[3H]QNB binding and contraction of rabbit colonic smooth muscle cells
Authors: Ringer M. J. , Hyman P. E. , Kao H. W. , Hsu C. T. , Tomomasa T. , Snape W. J.
Published: December 2017

We used radioligand binding and studies of cell contraction to characterize muscarinic receptors on dispersed smooth muscle cells from rabbit proximal and distal colon. Cells obtained after serial incubations in collagenase were used to measure binding of tritiated quinuclidinyl benzilate [( 3H]QNB). AT 37 degrees C, specific [3H]QNB binding was saturable and linearly related to cell number. Nonlinear regression analysis was used to determine the affinity of [3H]QNB for its receptor. In the distal colon the Kd was 60 pM, and the mean number of receptors was 1.2 X 10(6)/cell. Compared with cells from the distal colon, cells from the proximal colon had a lower affinity (Kd = 337 pM) but similar numbers of receptors. The concentrations of the antagonists atropine, secovorine, and pirenzepine, which were required for inhibition of 50% [3H]QNB binding (IC50), were 8, 5, and 870 nM, respectively, suggesting that the receptors are of the M2-muscarinic subclass. Hill coefficients for these agents were 1.1, 0.9, and 1.1, suggesting binding to a single receptor. The IC50 for the muscarinic agonists bethanechol and oxotremorine were 80 and 0.57 microM, respectively. Hill coefficients were 0.67 for both, suggesting more complex interactions involving receptors of different affinities. In studies of cell contraction, bethanechol stimulated a dose-dependent decrease in cell length with half the maximal contraction occurring at 100 pM. These results suggest that 1) contraction is mediated by binding of bethanechol to M2-muscarinic receptors and that 2) there are a large number of spare receptors in colonic smooth muscle.
Receptors for substance P on isolated intestinal smooth muscle cells of the guinea pig
Authors: Souquet J. C. , Bitar K. N. , Grider J. R. , Makhlouf G. M.
Published: December 2017

Two radioligands, 125I-labeled substance P (125I-SP) and 125I-labeled substance K (125I-SK), were used to characterize the kinetics and stoichiometry of binding of mammalian tachykinins [substance P (SP), substance K (SK), and neuromedin K (NK)] to smooth muscle cells isolated from the longitudinal muscle layer of guinea pig intestine. Specific binding of 125I-SP and 125I-SK was rapid, saturable, reversible, and temperature dependent. Binding attained 63-70% of steady-state binding within 1 min, coincidentally with the time of optimal contraction. The order of potency with which mammalian tachykinins and the SP antagonist, [D-Pro2, D-Trp7,9]SP, inhibited the binding of both radioligands was identical: SP greater than SK greater than NK greater than [D-Pro2, D-Trp7,9]SP, implying preferential interaction with a site that had highest affinity for SP. SK was 2-3 times, NK 3-4 times, and [D-Pro2, D-Trp7,9]SP 7-23 times less potent than SP (IC50 0.36 nM). Except for NK, the order of potency was similar to that for contraction of isolated muscle cells. The existence of binding sites with even higher affinity was suggested by the ability of muscle cells to contract in response to concentrations as low as 10(-13) M. These binding sites were not detectable at the concentration of radioligands used. It was concluded that a SP receptor is the only tachykinin receptor subtype present on intestinal muscle cells of the guinea pig.
Pancreatic polypeptide inhibits pancreatic enzyme secretion via a cholinergic pathway
Authors: Jung G. , Louie D. S. , Owyang C.
Published: December 2017

In rat pancreatic slices, rat pancreatic polypeptide (PP) or C-terminal hexapeptide of PP [PP-(31-36)] inhibited potassium-stimulated amylase release in a dose-dependent manner. The inhibition was unaffected by addition of hexamethonium but blocked by atropine. In contrast, PP(31-36) did not have any effect on acetylcholine- or cholecystokinin octapeptide-stimulated amylase release. In addition, when pancreatic slices were incubated with [3H] choline, PP(31-36) inhibited the potassium-evoked release of synthesized [3H] acetylcholine in a dose-dependent manner. The inhibitory action of PP was unaffected by adrenergic, dopaminergic, or opioid receptor antagonists. Thus PP inhibits pancreatic enzyme secretion via presynaptic modulation of acetylcholine release. This newly identified pathway provides a novel mechanism for hormonal inhibition of pancreatic enzyme secretion via modulation of the classic neurotransmitter function.
Role of rat intestinal brush-border membrane angiotensin-converting enzyme in dietary protein digestion
Authors: Yoshioka M. , Erickson R. H. , Woodley J. F. , Gulli R. , Guan D. , Kim Y. S.
Published: December 2017

The role of rat intestinal angiotensin-converting enzyme (ACE; E.C 3.4.15.1) in the digestion and absorption of dietary protein was investigated. Enzyme activity was associated with the brush-border membrane fraction, with the highest activity in the proximal to midregion of the small intestine. Preliminary enzyme characterization studies were carried out using purified brush-border membrane preparations. When a variety of N-blocked synthetic peptides were used as potential substrates for ACE, activity was highest with those containing proline at the carboxy terminal position. The hydrolytic rates observed with these prolyl peptides were comparable to those observed when major digestive peptidases of the brush-border membrane such as aminopeptidase N and dipeptidyl aminopeptidase IV were assayed. When isolated rat jejunum was perfused in vivo with solutions of Bz-Gly-Ala-Pro, the dipeptide Ala-Pro was the main hydrolytic product detected in the perfusates. Absorption rates of the constituent amino acids, alanine and proline, depended on the concentration of peptide perfused. Captopril, an active site specific ACE inhibitor, significantly inhibited hydrolysis and absorption of constituent amino acids from Bz-Gly-Ala-Pro. These results show that intestinal brush-border membrane ACE functions as a digestive peptidase in addition to its role as a regulator of biologically active peptides in other tissues.
Intact biliary excretion of gastrically administered prostaglandin F2 alpha in rats: developmental differences
Authors: Bedrick A. D. , Wells M. A. , Ford D. L. , Koldovsky O.
Published: December 2017

Tritium-labeled prostaglandin F2 alpha was administered via orogastric tube to bile duct-cannulated suckling and weanling rats to determine if maturational differences were present in the biliary excretion of prostaglandin F2 alpha and metabolites. Animals were killed 2 h after radioactivity administration. Characterization of radioactivity present in bile revealed age-related differences in biliary prostaglandin F2 alpha excretion. Suckling rats had a greater proportion of radioactivity migrating in chromatographic regions of greater polarity than prostaglandin F2 alpha. Compared with the weanling, a significantly greater amount of radioactivity cochromatographed with intact, unmetabolized prostaglandin F2 alpha (33.08 +/- 1.99 vs. 21.38 +/- 1.46). These results indicate that orogastrically administered prostaglandin F2 alpha can be absorbed from the gastrointestinal tract, transported to the liver, and subsequently excreted into bile and detected in an unmetabolized form in suckling and weanling rats. The enterohepatic circulation of milk-derived prostaglandin present in bile may contribute to the overall content of intestinal prostaglandins.
Characterization of gastrin receptors on guinea pig pancreatic acini
Authors: Yu D. H. , Noguchi M. , Zhou Z. C. , Villanueva M. L. , Gardner J. D. , Jensen R. T.
Published: December 2017

Recent studies have demonstrated gastrin receptors in some pancreatic tumors and that gastrin is a potent stimulant of pancreatic Na+-H+ exchange. In the present study we used 125I-labeled gastrin (125I-gastrin) to characterize gastrin receptors on guinea pig pancreatic acini. Binding of 125I-gastrin was temperature dependent, saturable, and specific for gastrin-related peptides. Analysis demonstrated a single class of receptors with high affinity for gastrin (Kd = 1.5 nM) and a binding capacity of 1 fmol/mg protein. Binding of 125I-gastrin was inhibited with the following relative potencies (Kd): cholecystokinin octapeptide (CCK-8) (0.35 nM) greater than gastrin-17-I = gastrin-34-I (1.5 nM) greater than pentagastrin (7 nM) greater than desulfated [des(SO3)]CCK-8 (28 nM) greater than CCK-4 (508 nM) and by the receptor antagonists CBZ-CCK-27-32-NH2 (3.5 microM) greater than proglumide analogue 10 (30 microM) greater than asperlicin (265 microM) greater than Bt2-guanosine 3',5'-cyclic monophosphate (828 micron). In contrast, for both stimulation of enzyme secretion and inhibition of binding of 125I-CCK-8 the relative potencies were CCK-8 much greater than des(SO3)CCK-8 greater than gastrin-17-I = gastrin-34-I greater than pentagastrin greater than CCK-4. For each receptor antagonist the dose-inhibition curve for gastrin-stimulated amylase release was superimpossible with that for CCK-8-stimulated amylase release. Gastrin-17-I at concentrations less than 0.1 microM did not potentiate carbachol or vasoactive intestinal peptide-stimulated amylase secretion and did not affect basal or stimulated adenosine 3',5'-cyclic monophosphate or 45Ca outflux.(ABSTRACT TRUNCATED AT 250 WORDS)
Effects of ethanol on parietal cell membrane phospholipids and proton pump function
Authors: Mazzeo A. R. , Nandi J. , Levine R. A.
Published: December 2017

The effects of ethanol on membrane phospholipids of isolated rabbit parietal cells (PC) and fundic glands (FG) were evaluated by using proton nuclear magnetic resonance spectroscopy (NMR). The most conspicuous resonance bands in PC and FG spectra at 3.2 and 0.9 ppm were due to the protons of +N(CH3)3 groups of phosphatidylcholine and, to a lesser degree, other phospholipid derivatives and terminal -CH3 groups for all phospholipids, respectively. Ethanol treatment (1 min) of PC and FG at 37 degrees C significantly increased the peak intensity of -CH3 groups in a dose-dependent manner, while no intensity change in PC and a lesser change in FG were observed for the +N(CH3)3 groups, suggesting a greater disorganizing effect of ethanol in the inner hydrophobic (-CH3) compared with the outer hydrophilic [+N(CH3)3] membrane moieties of phospholipids. Ethanol at concentrations greater than 5% (vol/vol) inhibited K+-stimulated paranitrophenyl phosphatase activity of intact PC, which correlated with inhibition of basal and histamine-stimulated aminopyrine uptake ratio by PC and FG. NMR is a sensitive, noninvasive probe to show ethanol-induced changes in intact PC lipid membrane organization without altering ultrastructural morphology.
Functionally distinct receptors for cholecystokinin and gastrin on dispersed chief cells from guinea pig stomach
Authors: Cherner J. A. , Sutliff V. E. , Grybowski D. M. , Jensen R. T. , Gardner J. D.
Published: December 2017

Caerulein, gastrin, and C-terminal fragments of cholecystokinin (CCK) varying in length from eight (CCK-8) to four (CCK-4) amino acids stimulate pepsinogen secretion from dispersed chief cells prepared from guinea pig stomach. C-terminal fragments of CCK containing fewer than four amino acids, even when tested at concentrations as high as 3 mM, do not stimulate pepsinogen secretion. The efficacies of gastrin and the various CCK-related peptides, coupled with the pattern of action of CCK receptor antagonists, indicate that chief cells from guinea pig stomach possess two functionally distinct classes of receptors, C-receptors and G-receptors. The C-receptors can be occupied by caerulein, CCK-8, CCK-7, des(SO3)CCK-8, or des(SO3)CCK-7, and occupation of C-receptors causes full stimulation of pepsinogen secretion. G-receptors can be occupied by gastrin I, gastrin II, CCK-6, CCK-5, or CCK-4, and occupation of G-receptors causes stimulation of pepsinogen secretion that is 60% of maximal.
Effect of somatostatin on basal and stimulated gastric secretion in the cod, Gadus morhua
Authors: Holstein B. , Cederberg C.
Published: December 2017

In vivo secretion of gastric acid and pepsin has been studied in pylorus-ligated cod. Basal acid output amounted to 100-150 mumol H+.kg-1.h-1 and pepsin secretion to 1 mg.kg-1.h-1. In response to bombesin nonapeptide (2.4 nmol.kg-1.h-1) and histamine (81 nmol.kg-1.h-1), acid secretion increased to approximately 200 and 600% of the basal level, respectively. Pepsin output was marginally affected by histamine but increased to approximately 3 and 15 times the basal level during treatment with bombesin and eledoisin (3.27 nmol.kg-1.h-1). Somatostatin (SS-14, 15 nmol.kg-1.h-1) inhibited basal acid secretion by 85%. It also inhibited the acid secretion during stimulation with bombesin (68%) and histamine (57%), but although the former effect could be explained by removal of the basal component, the latter could not. Basal pepsin secretion was not affected by SS-14. A slight inhibition (28%) of the peak pepsin response to eledoisin was demonstrated, and bombesin failed to stimulate pepsin secretion during treatment with SS-14. These results indicate that endogenous somatostatin, if present in the cod stomach, could play a role in the regulation of gastric secretion.
Somatostatin modulates cholinergic neurotransmission in canine antral muscle
Authors: Koelbel C. B. , van Deventer G. , Khawaja S. , Mogard M. , Walsh J. H. , Mayer E. A.
Published: December 2017

Somatostatin has been shown to inhibit antral motility in vivo. To examine the effect of somatostatin on cholinergic neurotransmission in the canine antrum, we studied the mechanical response of and the release of [3H]acetylcholine from canine longitudinal antral muscle in response to substance P, gastrin 17, and electrical stimulation. In unstimulated tissues, somatostatin had a positive inotropic effect on spontaneous phasic contractions. In tissues stimulated with substance P and gastrin 17, but not with electrical stimulation, somatostatin inhibited the phasic inotropic response dose dependently. This inhibitory effect was abolished by indomethacin. Somatostatin stimulated the release of prostaglandin E2 radioimmunoreactivity, and prostaglandin E2 inhibited the release of [3H]acetylcholine induced by substance P and electrical stimulation. Somatostatin increased the release of [3H]acetylcholine from unstimulated tissues by a tetrodotoxin-sensitive mechanism but inhibited the release induced by substance P and electrical stimulation. These results suggest that somatostatin has a dual modulatory effect on cholinergic neurotransmission in canine longitudinal antral muscle. This effect is excitatory in unstimulated tissues and inhibitory in stimulated tissues. The inhibitory effect is partially mediated by prostaglandins.
Laser-Doppler flowmetry for estimating liver blood flow
Authors: Arvidsson D. , Svensson H. , Haglund U.
Published: December 2017

Whether laser-Doppler flowmetry can be used to monitor liver blood flow was evaluated in a porcine model in which portal venous blood flow was followed indirectly by electromagnetic flowmetry applied to the superior mesenteric artery, and total hepatic venous outflow was measured directly by using an extracorporeal circuit. Hepatic venous outflow at rest was 23.5 +/- 5.7 ml.kg body wt-1.min-1. Occlusion of the hepatic arterial supply reduced hepatic laser-Doppler blood flow to 22%, but hepatic venous outflow only to 80%. Portal venous blood flow remained unchanged or increased slightly. Occlusion of the portal vein, on the other hand, decreased hepatic laser-Doppler blood flow values to 37% and hepatic venous outflow to 13%. Increased hepatic venous outflow pressure, caused by a positive end-expiratory pressure or elevation of the draining cannula, reduced flow and caused approximately equal changes in the three variables, as did reduced flow by step-wise bleeding. From these experiments in the pig it is concluded that laser-Doppler flowmetry on the liver surface clearly reflects relative changes of the total liver blood flow, as exemplified in this study, during venous stasis and bleeding. The technique is, however, more sensitive to blood flow changes in the hepatic artery as compared with the portal vein.
Ethanol-induced increase in portal blood flow: role of adenosine
Authors: Orrego H. , Carmichael F. J. , Saldivia V. , Giles H. G. , Sandrin S. , Israel Y.
Published: December 2017

The mechanism by which ethanol induces an increase in portal vein blood flow was studied in rats using radiolabeled microspheres. Ethanol (2 g/kg) by gavage resulted in an increase of 50-70% in portal vein blood flow. The ethanol-induced increase in portal blood flow was suppressed by the adenosine receptor blocker 8-phenyltheophylline [ethanol, 61.8 +/- 4.1 ml.kg-1.min-1; ethanol + 8-phenyltheophylline (0.2 mg.kg-1.min-1), 44.2 +/- 2.0 ml.kg-1.min-1; P less than 0.05]. By itself, 8-phenyltheophylline (0.2 mg.kg-1.min-1) was without effect on cardiac output or portal blood flow. Adenosine infusion resulted in a dose-dependent increase in portal blood flow with a maximal effect at a dose of 0.17 mg.kg-1.min-1 (control, 41.3 +/- 2.3; adenosine, 81.7 +/- 8.0 ml.kg-1.min-1; P less than 0.05). This adenosine-induced increase in portal blood flow was inhibited by 8-phenyltheophylline in a dose-dependent manner [adenosine, 81.7 +/- 8.0 ml.kg-1.min-1; adenosine + 8-phenyltheophylline (0.2 mg.kg-1.min-1), 49.8 +/- 6.6 ml.kg-1.min; P less than 0.05]. Both alcohol and adenosine significantly reduced preportal vascular resistance by 40% (P less than 0.02) and 60% (P less than 0.01), respectively. These effects were fully suppressed by 8-phenyltheophylline. It is concluded that adenosine is a likely candidate to mediate the ethanol-induced increase in portal vein blood flow. It is suggested that an increase in circulating acetate and liver hypoxia may mediate the effects of alcohol by increasing tissue and interstitial adenosine levels.
Autoregulatory escape from norepinephrine infusion: roles of adenosine and histamine
Authors: Crissinger K. D. , Kvietys P. R. , Granger D. N.
Published: December 2017

Stimulation of sympathetic fibers or infusion of norepinephrine (NE) into the superior mesenteric artery (SMA) leads to an initial decrease in intestinal blood flow, which is followed by a return of flow toward the base-line value (autoregulatory escape) despite continued nerve stimulation or NE infusion. Although the mechanisms responsible for “autoregulatory escape” have not been defined, accumulation of vasodilator metabolites is frequently invoked to explain this phenomenon. Inasmuch as histamine and adenosine exist in high concentrations in the intestinal mucosa and both are potent vasodilators, we examined the effects of chlorpheniramine (an H1 blocker) and adenosine deaminase (degrades adenosine) on autoregulatory escape from NE infusion. In autoperfused piglet intestinal preparations, we measured SMA blood flow and the arteriovenous oxygen difference during intra-arterial NE infusion before and after blockade with chlorpheniramine or adenosine deaminase. Adenosine deaminase pretreatment increased the peak vasoconstrictor and reduced the steady-state escape responses to NE infusion. Chlorpheniramine did not affect either the vasoconstrictor or escape responses. The oxygen uptake changes induced by NE infusion were not dramatically modified by either treatment. These results indicate that adenosine but not histamine is responsible for at least part of the escape of intestinal blood flow from NE infusion.
Giant migrating contractions of the canine cecum
Authors: Sarna S. K. , Prasad K. R. , Lang I. M.
Published: December 2017

We investigated cecal motor activity and its coordination with ileal and colonic motor activities in five conscious dogs instrumented with strain-gauge transducers. Both ileum and colon exhibited the usual migrating motor complexes (MMCs) that were independent of each other. Cecum had no motor complexes, but it periodically generated giant migrating contractions (GMCs) that originated at the apex and migrated to the cecum-colonic junction at a velocity of 13 +/- 1.3 cm/min. The amplitude of cecal GMCs was 1.82 +/- 0.26 and 3.37 +/- 0.39 times larger than that of contractions during ileal and colonic motor complexes, respectively (P less than 0.05). The mean duration and period of cecal GMCs were 45 +/- 4 s and 61.4 +/- 14.2 min, respectively, in the fasted state and 49 +/- 6 s and 68.5 +/- 13.5 min in the fed state, respectively (P less than 0.05). A total of 82 cecal GMCs was recorded in 71 h of fasted recording and 94 in 121 h of fed recording. Cecal GMCs were not coordinated with colonic MMCs. Intravenous motilin, but not morphine, initiated a premature cecal GMC. We concluded that cecum has a unique motor activity that is different from that of the colon and the ileum. The strong caudad GMCs of the cecum may periodically empty cecal contents into the colon.
Cholestasis and the interactions of sulfated glyco- and taurolithocholate with calcium
Authors: van der Meer R. , Vonk R. J. , Kuipers F.
Published: December 2017

To elucidate possible mechanisms of the differential cholestatic effect of sulfated glyco- and taurolithocholate (SGLC and STLC) the interactions of these bile acids with calcium were studied both in vitro and in vivo. In vitro, SGLC was readily precipitated by Ca in physiological concentrations (less than 5 mM). It appeared that only micelles of SGLC were precipitated and that both the sulfate and the carboxylate anionic groups participated in Ca binding. Conjugation with taurine almost completely prevented this Ca-dependent precipitation. Intravenous infusion of SGLC (12 mumol/h) in unanesthetized rats with permanent biliary drainage induced cholestasis, which was preceded by the formation of a biliary Ca-SGLC precipitate, suggesting a causal relationship. Taurine conjugation prevented this biliary precipitation as well as cholestasis. Furthermore, we studied in vitro the binding of the sulfated bile acids to insoluble calcium phosphate, simulating interactions with Ca at the intestinal level. Compared with STLC, micellar SGLC was bound with much higher affinity. This high-affinity binding was probably due to the involvement of both the sulfate and the carboxylate group. This might also explain that the apparent critical micellar concentration of SGLC, in contrast to that of STLC, was extremely Ca dependent. The present findings indicate that 1) interaction with calcium might be responsible for the differential cholestatic properties of SGLC and STLC and 2) binding of sulfated lithocholic acid conjugates to insoluble calcium phosphate in the intestinal lumen is likely to prevent their accumulation in the enterohepatic circulation.
Effect of GABA on basal and vagally mediated gastric acid secretion and hormone release in dogs
Authors: Thirlby R. C. , Stevens M. H. , Blair A. J. , Petty F. , Crawford I. L. , Taylor I. L. , Walsh J. H. , Feldman M.
Published: December 2017

To stimulate peripheral gamma-aminobutyric acid (GABA) receptors, GABA, which does not cross the blood-brain barrier, was administered to dogs with vagally innervated gastric fistulas at intravenous doses of 0, 0.66, 2, 6, 18, and 54 micrograms.kg-1.min-1. Mean gastric acid output increased from zero basally to 3.0 +/- 1.4 mmol/h during infusion of 54 micrograms.kg-1.min-1 GABA. Plasma somatostatin-like immunoreactivity decreased significantly below basal levels during infusion of 54 micrograms.kg-1.min-1 GABA (P less than 0.05). To stimulate central nervous system GABA receptors as well as peripheral GABA receptors, progabide, a GABA-receptor agonist, which readily crosses the blood-brain barrier, was injected intravenously. Mean acid output was 3.5 +/- 1.3 mmol/h after 20 mg/kg progabide and 0.6 +/- 0.5 mmol/h after its vehicle (P less than 0.05). Basal serum gastrin concentration increased significantly after progabide injection. Acid output during insulin-induced hypoglycemia was inhibited 59% by 30 mg/kg intravenous progabide. Progabide infusion also diminished or abolished circulating gastrin, somatostatin, and pancreatic polypeptide responses during insulin-induced hypoglycemia (P less than 0.05). Further studies were performed in dogs with a gastric fistula and a vagally denervated Heidenhain pouch to confirm that GABA-receptor stimulation affects acid secretion via peripheral pathways. Intravenous injection of baclofen (0.5 mg/kg), a GABAB-receptor agonist, increased acid secretion significantly from the gastric fistula and the Heidenhain pouch. These studies suggest that GABA may play a role in regulating gastric acid secretion and gastrointestinal and pancreatic endocrine function by both central and peripheral mechanisms.
Starch and glucose oligosaccharides protect salivary-type amylase activity at acid pH
Authors: Rosenblum J. L. , Irwin C. L. , Alpers D. H.
Published: December 2017

Salivary-type amylase may significantly contribute to duodenal starch hydrolysis in exocrine pancreatic insufficiency, provided that gastric inactivation does not occur. We investigated the effect of starch and its hydrolytic products, therefore, on salivary amylase activity in vitro at low pH. When incubated at pH 3 in the presence of 1% starch, 56% of the initial activity of amylase purified from saliva remained after 60 min at 37 degrees C compared with only 6% without starch. Similar protection of amylolytic activity was observed using human milk, which also contains a salivary-type amylase. In addition, partially hydrolyzed starch protected salivary amylase activity at pH 3, and purified glucose oligomers ranging in length from two to seven glucose molecules protected amylase in a concentration-dependent manner. Lactose, sucrose, and glucose, however, were ineffective in sparing amylase. Starch protected amylase activity even in the presence of pepsin, and maltotriose conferred striking protection below pH 3. These studies indicate that salivary-type isoamylases are protected in a simulated gastric environment by substrates of amylase as well as its end products of hydrolysis.
Effects of leukotriene C4 on pancreatic secretion and circulation in dogs
Authors: Konturek S. J. , Pawlik W. , Czarnobilski K. , Gustaw P. , Jaworek J. , Beck G. , Jendralla H.
Published: December 2017

In the present study the effects of leukotriene C4 (LTC4) on exocrine pancreatic secretion and pancreatic blood flow were determined. LTC4 given intravenously in various doses ranging from 0.35 to 2.8 nmol.kg-1.h-1 in conscious dogs caused a dose-dependent inhibition of pancreatic HCO-3 and protein responses to exogenous hormones such as secretin, cholecystokinin octapeptide (CCK-8), and bombesin and to endogenous stimulants including meat feeding and duodenal perfusion with oleate. In tests with pancreatic secretion induced by secretin plus CCK, maximal inhibition by LTC4 occurred at a dose of 1.4 nmol.kg-1.h-1 and reached approximately 70% of the control value for HCO-3 output and 45% for protein output. In tests with separate secretin- or CCK-induced secretion, maximal inhibition occurred at a dose of 1.4 nmol.kg-1.h-1 and reached 38 and 66% of the control HCO-3 and protein secretion, respectively. The same dose of LTC4 reduced the postprandial HCO-3 secretion by approximately 80% and protein output by approximately 70%. After administration of indomethacin, the pancreatic secretion declined, but the inhibitory effects of LTC4 remained unchanged. Pancreatic tissue generated two to three times more LTC4 than the gastrointestinal mucosa, and indomethacin caused further increase in this generation, suggesting that LTC4 may contribute to indomethacin-induced pancreatic inhibition. (ABSTRACT TRUNCATED AT 250 WORDS)
Neurohumoral control of villous motility
Authors: Womack W. A. , Mailman D. , Kvietys P. R. , Granger D. N.
Published: December 2017

A quantitative videomicroscopic method was used to examine neurohumoral control of villous motility. Intraduodenal instillation of saline, 0.4% hydrochloric acid, or acidified predigested food did not cause a significant change in villous contraction frequency in an isolated segment of jejunum. Villous motility in the jejunum of fed dogs, from which the chyme had been removed, was not greater than that in fasted dogs (2.9 +/- 0.3 vs. 3.4 +/- 0.5 contractions/min). Furthermore, acid extracts of the duodenal mucosa did not produce an increase in jejunal villous motility when injected intravenously. These data argue against the existence of a humoral stimulant of villous motility (villikinin). Vagotomy caused only a small (20%) and transient (10 min) decline in villous motility. Vagal stimulation at 5, 10, and 20 Hz caused villous motility to increase by 24 +/- 7, 23 +/- 9, and 32 +/- 10%, respectively. Atropine blocked the effects of vagal stimulation. Section of the periarterial (sympathetic) nerves did not alter villous contractile activity. Stimulation of the periarterial nerves at 5, 10, and 20 Hz caused villous contraction frequency to decline by 41 +/- 5, 45 +/- 5, and 38 +/- 10%, respectively. This inhibition appears to involve both alpha- and beta-adrenergic receptors and a reduction in blood flow. Neither atropine, alpha-blockade, nor beta-blockade produced a sustained alteration in basal contraction frequency.
Characterization of a new CCK antagonist, L364,718: in vitro and in vivo studies
Authors: Louie D. S. , Liang J. P. , Owyang C.
Published: December 2017

In this study we examined a novel, orally effective, nonpeptidal cholecystokinin (CCK) antagonist, 3S(-)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepine-3-yl) -1H-indole-2-carboxamide (L364,718) on CCK-induced amylase release. We used isolated rat pancreatic acini and incubated them with CCK-8 with or without various CCK receptor antagonists. L364,718 (3-100 nM), proglumide (1-10 mM), and the proglumide derivative CR1409 (1-30 microM) each caused a progressive rightward shift in the CCK-8 dose-response curve without a change in maximal amylase secretion. Plotting the data according to the method of Schild indicated competitive inhibition. The Kis for L364,718, CR1409, and proglumide were 0.25 nM, 0.15 microM, and 0.5 mM, respectively. Thus L364,718 was 600-fold more potent than CR1409 and 2,000,000-fold more potent than proglumide in inhibiting CCK-8-induced amylase release. Inhibition of 125I-Bolton-Hunter-CCK-8 binding to acini by these receptor antagonists had a similar rank potency. L364,718 was tested against other pancreatic exocrine secretagogues and was effective against agonists that only act through the CCK receptor. In rats, diversion of pancreatic juice from the duodenum stimulated pancreatic amylase output and elevated plasma CCK levels. The stimulated amylase secretion was blocked by intravenous administration of L364,718. Furthermore, extracted plasma CCK from rats with diversion of pancreatic juice from the duodenum stimulated amylase release from pancreatic acini, and this stimulation was blocked by addition of L364,718.(ABSTRACT TRUNCATED AT 250 WORDS)
Ethanol-induced increase in portal blood flow: role of acetate and A1- and A2-adenosine receptors
Authors: Carmichael F. J. , Saldivia V. , Varghese G. A. , Israel Y. , Orrego H.
Published: December 2017

The increase in portal blood flow induced by ethanol appears to be adenosine mediated. Acetate, which is released by the liver during ethanol metabolism, is known to increase adenosine levels in tissues and in blood. The effects of acetate on portal blood flow were investigated in rats using the microsphere technique. The intravenous infusion of acetate (7-250 mumol.kg-1.min-1) resulted in vasodilation of the preportal vasculature and in a dose-dependent increase in portal blood flow [control, 39.1 +/- 2.6 ml.kg-1.min-1; acetate (250 mumol.kg-1. min-1), 68.7 +/- 4.0 ml.kg-1.min-1]. This acetate-induced increase in portal blood flow was suppressed by the adenosine receptor blocker, 8-phenyltheophylline. Using the A1-adenosine receptor agonist N-6-cyclohexyl adenosine and the A2-agonist 5'-N-ethylcarboxamido adenosine, we demonstrate that the effect of adenosine on the preportal vasculature is mediated by the A2-subtype of adenosine receptors. In conclusion, these data support the hypothesis that the increase in portal blood flow after ethanol administration results from a preportal vasodilatory effect of adenosine formed from acetate metabolism in extrahepatic tissues.
H+-dependent calcium uptake into an IP3-sensitive calcium pool from rat parotid gland
Authors: Thevenod F. , Schulz I.
Published: December 2017

In permeabilized parotid cells and in isolated membrane vesicles from parotid endoplasmic reticulum (ER), Mg-ATP-dependent Ca2+ uptake was measured using a Ca2+-specific macroelectrode and 45Ca2+, respectively. Mg-ATP-dependent Ca2+ uptake was inhibited by vanadate (2 x 10(-3) mol/l) by approximately 45% in permeabilized cells and by approximately 70% in membrane vesicles from ER during the initial 10 min. After this lag phase, Ca2+ uptake increased and low steady-state free [Ca2+] of approximately 3 x 10(-7) mol/l was still reached in presence of vanadate within 30-40 min. Subsequent addition of inositol 1,4,5-trisphosphate (IP3) caused a similar Ca2+ release compared with control. This indicates that in presence of vanadate an IP3-sensitive Ca2+ pool was filled. However, when protonophores, such as nigericin or carbonyl cyanide-m-chlorophenylhydrazone, were added in addition to vanadate, this low steady-state free [Ca2+] was not reached. 45Ca2+ uptake was reduced by approximately 70% within 60 min, and IP3 did not cause 45Ca2+ release when given subsequently, indicating that filling of an IP3-sensitive Ca2+ pool was prevented. Mg-ATP-driven H+ uptake into ER vesicles, as estimated with acridine orange, was abolished by protonophores and by the H+-ATPase blockers N-ethylmaleimide and Dio 9 but was unaltered by vanadate. Preincubation of ER vesicles in a medium without Ca2+, but with vanadate and with Mg-ATP to generate an H+ gradient, allowed demonstration of 45Ca2+ uptake from a medium that did not contain ATP. The cation sequence in absence of vanadate for support of Mg-ATP-dependent 45Ca2+ uptake was K+ greater than Na+ greater than Li+ = choline+ and, in presence of vanadate, was choline+ greater than Li+ = Na+ greater than K+. A preformed H+ gradient dissipated more rapidly in presence of K+ compared with choline+, probably due to an intrinsic K+ permeability of ER membrane. Our data indicate that both a Ca2+ and a H+ pump are located in a compartment of ER that is also sensitive to IP3. Ca2+ uptake is coupled to an H+ gradient that is generated by the H+ pump and most likely occurs via Mg-ATP-driven Ca2+-H+ countertransport but to some extent can also operate in absence of ATP at the expense of the H+ gradient.
Role of glucagon in intestinal hyperemia associated with early experimental diabetes mellitus
Authors: Yrle L. F. , Smith J. K. , Benoit J. N. , Granger D. N. , Korthuis R. J.
Published: December 2017

The role of glucagon as a blood-borne mediator of the intestinal hyperemia associated with experimental diabetes mellitus was assessed in anesthetized fasted (18-24 h) rats 4 wk after the administration of streptozotocin (65 mg/kg body wt) or its vehicle. Selective removal of pancreatic glucagon from the circulation was accomplished by the intravenous administration of a highly specific glucagon antiserum. Blood flow to the gastrointestinal tract and kidneys was measured with radioactive microspheres using the reference sample technique. Blood flows were increased by at least 60% in each segment of the gastrointestinal tract of diabetic animals compared with control rats. Glucagon antiserum had no effect on blood flows in the gastrointestinal tract of control animals. However, the antiserum produced a significant reduction in blood flow to the stomach (26%), duodenum (25%), jejunum (12%), and kidneys (16%) in diabetic rats. There was no change in blood flow to the ileum or colon of diabetic animals with antiserum administration. The results of this study support the hypothesis that glucagon mediates a portion of the hyperemia noted in the stomach, duodenum, and jejunum. However, glucagon does not appear to play a role in the genesis of the hyperemia noted in more distal segments of the gastrointestinal tract (ileum and colon). A possible role for glucagon in the maintenance of renal blood flow in diabetic rats is suggested.
SOD prevents damage and attenuates eicosanoid release in a rabbit model of necrotizing enterocolitis
Authors: Miller M. J. , McNeill H. , Mullane K. M. , Caravella S. J. , Clark D. A.
Published: December 2017

Necrotizing enterocolitis (NEC) was produced in anesthetized rabbits by transmural injection of intestinal loops with an acidified solution of casein and calcium gluconate, mimicking the luminal milieu of afflicted neonates. Intravenous infusion of superoxide dismutase (SOD) 15 min after NEC induction prevented intestinal damage. In ex vivo perfused intestinal loops, we determined the sites of eicosanoid release and their contribution to the vascular effects of N-formyl-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) in damaged and SOD-salvaged intestine. The vascular effluent was the primary site of stimulated eicosanoid release. The vascular responses to fMLP (vasoconstriction) and PAF (vasodilation) were not altered by SOD, although vascular resistance was higher in the SOD group. SOD treatment attenuated 1) transmural fluid shifts in ex vivo perfused intestinal preparations, an index of vascular permeability, 2) fMLP-induced prostaglandin E2, 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha), and leukotriene B4 (LTB4) release, and 3) PAF-induced release of 6-keto-PGF1 alpha and LTB4. Stimulated thromboxane B2 release was not altered by SOD. Thus NEC can be established by a luminal insult that causes local generation of free radicals and exaggerated release of prostaglandins and leukotrienes.
Histamine H2 receptor stimulation increases gastric emptying in monkeys
Authors: Dubois A. , Nompleggi D. , Castell D. O.
Published: December 2017

We studied the effect of the specific H2 receptor agonist dimaprit on gastric emptying using a dye dilution technique in five chair-adapted rhesus monkeys to determine simultaneously gastric emptying and H+ secretion. Continuous subcutaneous injections of either saline alone or dimaprit (15, 30, 60, 120, and 240 nmol.kg-1.min-1) were given in random order on separate days. Both fractional emptying rate and H+ output were significantly increased by higher doses of dimaprit, with resulting twofold concurrent increase of intragastric H+ concentration. Because intragastric administration of H+ decreases gastric emptying and secretion, we evaluated the possibility that the stimulatory action of a histamine H2 agonist was underestimated when gastric pH was greater during control than after dimaprit. When exogenous HCl was added to the stomach to achieve H+ concentrations similar to those measured after dimaprit, both fractional emptying and H+ secretion were decreased twofold. When these "acidified control values" were used as a point of comparison, the stimulatory effect of dimaprit on both fractional emptying and gastric secretion was greater, suggesting that the stimulatory actions of dimaprit are underestimated if one does not take into account the differences in intragastric H+ concentrations.
Influence of distension on absorption and villous structure in rat jejunum
Authors: Holzheimer G. , Winne D.
Published: December 2017

In urethan-anesthetized rats the appearance rates of urea (U), antipyrine (A), and alpha-methyl-D-glucoside (MG) in the venous blood of perfused jejunal segment were measured in the undistended state and after elevation of the intraluminal pressure up to 10 cmH2O. Serosal and inner cylindrical surface area of the jejunal segment were enlarged by maximally 100 and 150%, respectively. The absorption rates, however, increased only by 34 (U), 28 (A), and 26% (MG). The increase of the supravillous diffusion resistance contributed only partially to this effect. The "cylindrical" permeability coefficient (Pcyl, average permeability coefficient related to inner cylindrical area, neglecting villous structure) decreased by 39, 57, and 50%, respectively. Due to circular stretching, broad intervillous spaces were formed that covered finally approximately 40% of the mucosal surface area. The additional intervillous diffusion resistance in these spaces was more effective than the absorption through the lateral surface of the villi. Thus the overall permeability of the mucosa, i.e., Pcyl, was reduced. Calculations based on a simplified model of the mucosa confirmed the experimental results.
Cholecystokinin in pig plasma: release of components devoid of a bioactive COOH-terminus
Authors: Cantor P. , Rehfeld J. F.
Published: December 2017

Using radioimmunoassays specific for different sequences of cholecystokinin (CCK), we studied the intestinal release of CCK in pigs. After stimulation by intraduodenal infusion of HCl, plasma CCK concentrations, measured with an antiserum specific for the sulfated, bioactive sequence of CCK, increased from 1.4 +/- 0.7 to 42.7 +/- 11.7 pM in portal plasma and from 0.5 +/- 0.3 to 12.3 +/- 1.5 pM in peripheral plasma. The concentrations measured with an antiserum specific for the NH2-terminal sequence 5-10 of CCK-33 were considerably higher, increasing from 64 +/- 17 to 139 +/- 14 pM in portal plasma and from 69 +/- 7 to 102 +/- 9 pM in peripheral plasma. Chromatography suggested that the NH2-terminal immunoreactivity consisted of large CCK fragments devoid of the bioactive COOH-terminal octapeptide, i.e., desocta and/or desnona CCK-58, CCK-39, and CCK-33. The bioactive forms in both portal and peripheral plasma comprised CCK-58-, CCK-33-, CCK-22-, CCK-12-, and CCK-8-like forms. Generally, the CCK-22-like component predominanated, but although CCK-8 was more abundant than CCK-33 in portal plasma, these forms occurred in equal amounts in peripheral plasma. Large amounts of NH2-terminal immunoreactivity were also found in the venous effluent of the isolated perfused duodenum after stimulation with gastrin-releasing peptide; the venous perfusate contained mainly CCK-22- and CCK-8-like material, which together constituted greater than 80% of the bioactive CCK components. Both duodenal and jejunal mucosa contained components resembling NH2-terminal fragments as well as large amounts of intact CCK-58 and CCK-33.(ABSTRACT TRUNCATED AT 250 WORDS)
Intracisternal injection of a TRH analogue stimulates gastric luminal serotonin release in rats
Authors: Stephens R. L. , Tache Y.
Published: December 2017

Intracisternal injection of TRH analogue, RX 77368, which has been previously shown to induce a vagal-dependent stimulation of gastric acid secretion and contractility dose-dependently (10-300 ng) stimulated serotonin release into the gastric lumen of conscious pylorus-ligated rats. The stimulation of gastric acid and serotonin release occurred 15 and 30 min, respectively, after intracisternal injection of RX 77368 (100 ng) and lasted for over 2 h, as measured in urethan-anesthetized rats with gastric fistula. Cervical vagotomy and atropine pretreatment (0.1 mg/kg) completely abolished the RX 77368-stimulated gastric serotonin and acid output in acute gastric fistula rats, whereas in pylorus-ligated rats atropine pretreatment markedly reduced RX 77368-stimulated gastric fluid volume and serotonin output, but not serotonin concentration. Systemic administration of bethanechol chloride, histamine, or pentagastrin, at doses that elicited gastric acid secretory responses similar to that of intracisternal TRH analogue, produced little or no intraluminal serotonin release. These results demonstrate that intracisternal injection of TRH analogue induced a marked and sustained release of serotonin into the gastric lumen that is mediated by vagal-dependent mechanisms unrelated to enhanced acid secretion. Activation of muscarinic receptors appears to be involved, but the relative ineffectiveness of bethanechol in this system suggests that (an)other vagally mediated neurochemical event(s) stimulated by intracisternal RX 77368 acts in conjunction with muscarinic receptor activation to produce intraluminal serotonin release. The functional significance of serotonin release into the gastric lumen in relation with intracisternal TRH analogue-induced alterations of gastric secretion, motility and blood flow, and lesion formation remains to be elucidated.
Functional VIP receptors in the human mucus-secreting colonic epithelial cell line CL.16E
Authors: Laburthe M. , Augeron C. , Rouyer-Fessard C. , Roumagnac I. , Maoret J. J. , Grasset E. , Laboisse C.
Published: December 2017

A polarized human clonal intestinal cell line exhibiting mucus secretion (Cl.16E) was used to study the expression and function of vasoactive intestinal peptide (VIP) receptors in mucus-secreting cells. Cl.16E cells expressed one class of receptors with a KD of 0.13 nM and a capacity of 67 fmol/mg protein. Covalent labeling of receptors followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed a receptor protein with a Mr of 63,000 in Cl.16E cells. VIP stimulated adenylate cyclase activity in membranes from Cl.16E cells with an ED50 of 0.06 nM. In conditions where carbachol stimulated mucin secretion from filter-grown Cl.16E cells, VIP, dibutyryl adenosine 3',5'-cyclic monophosphate (DbcAMP), or forskolin did not alter basal secretion. However, VIP strongly potentiated carbachol-induced mucin secretion, since carbachol alone and VIP plus carbachol induced a 1.6- and 3.6-fold increase of mucin secretion above basal, respectively. This potentiating effect of VIP was mimicked by DbcAMP or forskolin. It was observed for VIP concentrations in the 0.1-100 nM range (ED50, 2 nM). VIP elicited a dramatic increase of intracellular cAMP levels in filter-grown Cl.16E cells with a dose-response curve (ED50, 4 nM) very similar to that observed for the modulation of mucin secretion. These studies suggest that the clonal cell line Cl.16E may be an invaluable cellular model for evaluating the neurohormonal control of mucus secretion.
Effects of 1,25(OH)2D3 on enterocyte basolateral membrane Ca transport in rats
Authors: Favus M. J. , Tembe V. , Ambrosic K. A. , Nellans H. N.
Published: December 2017

One, twenty-five dihydroxyvitamin D3 [1,25(OH)2D3], commonly known as calcitriol, stimulates intestinal Ca absorption through increased activity of a cellular transport process. To determine whether transcellular Ca transport involves energy-dependent Ca efflux across enterocyte plasma membrane in vitamin D-sufficient rats, in vitro bidirectional Ca fluxes were measured under short-circuited conditions across proximal duodenum from rats fed diets adequate in vitamin D and containing a normal Ca diet (NCD), a low Ca diet (LCD), or fed NCD and injected with 50 ng of 1,25(OH)2D3 daily for 4 days before study. LCD or 1,25(OH)2D3 increased Ca net flux [Jnet, mucosal-to-serosal flux minus the serosal-to-mucosal flux] by increasing Ca mucosal-to-serosal flux (Jm----s) (mean +/- SE, NCD vs. LCD vs. 1,25(OH)2D3, 16 +/- 4 vs. 179 +/- 18 vs. 82 +/- 21 nmol.cm-2. h-1, P less than 0.0001). Initial ATP-dependent Ca uptake rates by duodenal basolateral membrane vesicles (BLMV) was greater in vesicles from rats fed NCD compared with LCD and not different from NCD injected with 1,25(OH)2D3. These studies suggest that in vitamin D-replete animals, 1,25(OH)2D3 increases epithelial Ca Jm----s by mechanisms that do not involve ATP-dependent BLM Ca efflux. ATP-dependent Ca exit from the cell under these conditions may play a role in intracellular Ca homeostasis rather than Ca absorption.