bluuebunny/crossref_metadata_2025 huggingface dataset:
Calcium flux in vivo in the rat duodenum and ileum during pregnancy and lactation
Authors: Toraason M.
Published: December 2017

Intestinal calcium transport was examined in nonpregnant, pregnant, lactating, and 100 ng 1,25-dihydroxyvitamin D3-dosed rats. Solutions containing 150 mM NaCl, 1 mM Ca, and 0.1 microCi 45Ca were recirculated in vivo through the duodenum and ileum to determine calcium efflux from the lumen, influx into the lumen, and net absorption. In the duodenum of the nonpregnant control rats, efflux of calcium was greater than influx, resulting in net absorption of calcium. In the ileum, calcium efflux and influx were equivalent and there was little net movement of calcium. 1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] significantly increased efflux and net absorption of calcium in the duodenum without affecting influx. In the ileum 1,25(OH)2D3 had no effect on calcium flux. During pregnancy and lactation there was a marked increase in mucosal tissue, and expression of transport data per centimeter of intestine gave different results than transport expressed per gram of dry mucosal tissue. Per centimeter of intestine, duodenal efflux and net absorption, but not influx, increased significantly during pregnancy and to greater extent during lactation. Per gram of dry mucosa, duodenal efflux and net absorption were increased significantly only during lactation, and the increase was comparable with that observed in 1,25(OH)2D3-dosed rats. In the ileum of pregnant and lactating rats, efflux and influx expressed per centimeter of intestine increased to approximately the same extent, and net movement of calcium remained unchanged from the control level. Expression of ileal transport data per gram of dry mucosa resulted in no significant difference in calcium movement during pregnancy and lactation.(ABSTRACT TRUNCATED AT 250 WORDS)
Action of natural glucagon on pancreatic acini: due to contamination by previously undescribed secretagogues
Authors: Pandol S. J. , Sutliff V. E. , Jones S. W. , Charlton C. G. , O'Donohue T. L. , Gardner J. D. , Jensen R. T.
Published: December 2017

In dispersed acini from guinea pig pancreas, natural glucagon caused a two- to threefold increase in amylase secretion, and this natural glucagon-induced increase was augmented by theophylline. Natural glucagon also caused a sixfold increase in cellular cAMP but did not alter cellular cGMP or outflux of 45Ca. Natural glucagon caused detectable changes in cAMP and amylase secretion at a concentration of 1 microM, half-maximal stimulation at 10 microM, and maximal stimulation at 100 microM. Natural glucagon potentiated the increase in enzyme secretion caused by secretagogues that act by causing mobilization of cellular calcium but did not alter the increase in enzyme secretion caused by secretagogues that increase or mimic the action of cellular cAMP. At concentrations up to 100 microM, natural glucagon did not inhibit binding of 125I-vasoactive intestinal peptide. The potency with which glucagon stimulated amylase release and augmented the increase in amylase release caused by cholecystokinin or carbachol was the same with acini from rat or mouse pancreas as it was with acini from guinea pig pancreas. Biologically active synthetic glucagon did not increase enzyme secretion. On reverse-phase, high-pressure liquid chromatography of natural glucagon, the ability to stimulate enzyme secretion migrated differently from the glucagon. Natural glucagon, at concentrations that stimulated cAMP accumulation, did not react with vasoactive intestinal peptide or secretin radioimmunoassays. Neither insulin nor pancreatic polypeptide, which are known to contaminate natural glucagon preparations, increased enzyme secretion from pancreatic acini. Trypsin treatment abolished the ability of natural glucagon to increase enzyme secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
Interdigestive contractile patterns of the ileum in dogs
Authors: Siegle M. L. , Ehrlein H. J.
Published: December 2017

The aim of this study is to elucidate the nature of ileal interdigestive contractile patterns by the computerized analysis of the contraction spread and by videofluoroscopy. Conscious dogs equipped with closely spaced strain-gauge force transducers were used. Two patterns of repetitive, phasic contractions were recorded, migrating clusters and phase IIIs; both patterns consisted of repetitive, propagated contractions. Both patterns migrated aborad by sequential movement of contraction waves down the bowel. Consequently, the rate of migration of either of the entire patterns was slower than the propagation velocity of constituent, individual contraction waves. Both patterns differed in several parameters, especially the propagated contractions of the clusters spread over shorter distances (1.47 +/- 0.4 cm) than those of phase III (4.65 +/- 0.99 cm). Compared with these complex patterns, propagating power contractions represented single contractions that propagated aborad at the same velocity as the contraction waves of the complex patterns. All three patterns propelled luminal contents distally.
Glycine-extended progastrin processing intermediates: accumulation and cosecretion with gastrin
Authors: Sugano K. , Park J. , Dobbins W. O. , Yamada T.
Published: December 2017

Glycine-extended intermediates of peptide processing serve as substrates for carboxyl-terminal amidation, hence activation, of many brain-gut peptides. To explore the dynamics of accumulation and secretion of these important intermediates we utilized primary cultures of canine antral mucosal G-cells as a model system. Glycine-extended progastrin processing intermediates (G-Gly) accumulated rapidly in G-cells cultured in ascorbate-deficient media, exhibiting a fourfold increase over a 51-h culture period, while gastrin content fell to less than half of the initial level. In contrast, G-cells cultured in ascorbate-supplemented media accumulated G-Gly at a relatively low rate, while gastrin was preserved at a higher level. Under either condition, G-Gly and gastrin were progressively released into the culture media. The release of both immunoreactivities could be stimulated by bombesin and inhibited by somatostatin in similar fashion. By electron microscopy, the cultured G-cells exhibited no ultrastructural alterations. These data suggest that 1) the cellular homeostasis of G-Gly is regulated by the activity of an ascorbate-dependent amidation enzyme similar to one previously described in pituitary tissues, 2) carboxyl-terminal amidation is not an obligatory step for secretion of gastrin, and 3) the proportions of gastrin and G-Gly cosecreted from G-cells reflect their proportional accumulation within G-cell secretory granules. The physiological relevance of the released G-Gly has yet to be determined.
Hybrid blood flow probe for simultaneous H2 clearance and laser-Doppler velocimetry
Authors: DiResta G. R. , Kiel J. W. , Riedel G. L. , Kaplan P. , Shepherd A. P.
Published: December 2017

To perform two independent regional blood flow measurements in tissue volumes of similar dimensions, we designed a hybrid blood flow probe capable of measuring regional perfusion by both laser-Doppler velocimetry (LDV) and H2 clearance. The probe consisted of two fiber-optic light guides to conduct light between the surface of tissue of interest and a laser-Doppler blood flowmeter. Also contained within the probe were a platinum 25-microns H2-sensing electrode and a 125-microns H2-generating electrode. The probe can thus be used to measure local perfusion with H2 clearance. The H2 can either be inhaled or can be generated electrochemically at the locus of interest. Evaluation of the probe in the canine gastric mucosa indicated 1) that the relationship between mucosal flow measurements made simultaneously with H2 clearance and LDV was highly significant and linear and 2) that H2 clearance could potentially be used to calibrate the laser-Doppler blood flowmeter in absolute units. The methods of constructing the flow probes are discussed in detail.
gamma-Aminobutyric acid stimulates acid secretion from the isolated guinea pig stomach
Authors: Tsai L. H. , Taniyama K. , Tanaka C.
Published: December 2017

gamma-Aminobutyric acid (GABA) content was measured, and the effect of GABA on acid secretion was studied using the everted preparation of isolated guinea pig stomachs. GABA contents in the mucosa layer and the remaining layer were 20-24 nmol/g tissue and 34-42 nmol/g tissue, respectively. GABA at 10(-6) to 3 X 10(-5) M induced acid secretion, and the maximum secretion was obtained at 3 X 10(-5) M, that is approximately 1.6-fold of the spontaneous secretion and approximately half of the amount secreted by histamine at 3 X 10(-4) M. The GABA-induced acid secretion was inhibited by bicuculline, scopolamine, pirenzepine, proglumide, and tetrodotoxin, but not by cimetidine. Muscimol (3 X 10 to 10(-5) M), but not baclofen, induced acid secretion in a concentration-dependent manner. The responses to GABA and muscimol were antagonized by bicuculline. Scopolamine and tetrodotoxin completely inhibited the acid secretion induced by low concentrations of GABA and muscimol and to some extent the response induced by high concentrations of muscimol. All these results indicate that GABA induces acid secretion via the A type of GABA receptor, probably located mainly on the cholinergic neurons and partially on the nonneuronal cells in the guinea pig stomach.
Somatostatin and muscarinic inhibition of canine enteric endocrine cells: cellular mechanisms
Authors: Barber D. L. , Gregor M. , Soll A. H.
Published: December 2017

Using a recently developed canine primary enteric endocrine cell culture system, we have investigated the role of adenosine 3',5'-cyclic monophosphate (cAMP) in mediating the release of neurotensin and enteroglucagon. Epinephrine-stimulated peptide release was concomitant with an increase in cAMP accumulation. Carbachol and somatostatin (SRIF) markedly inhibited the epinephrine effect on both peptide release and cAMP content. The addition of 3-isobutyl-1-methylxanthine potentiated epinephrine-stimulated peptide release without altering the relative inhibition by carbachol and SRIF, suggesting that these agents did not inhibit endocrine cell function by increasing phosphodiesterase activity. To determine the role of cAMP production in mediating inhibition of peptide release, cells were incubated with the bacterial toxin, pertussis toxin (PT). In cultures pretreated with PT, carbachol inhibition of both peptide release and cAMP accumulation was completely reversed. In contrast, SRIF inhibition of cAMP content was completely reversed after PT treatment, but inhibition of peptide release was only partially reversed. Additionally, toxin treatment only partially reversed SRIF inhibition of forskolin- and calcium ionophore-stimulated peptide release. These data suggest that muscarinic cholinergic inhibition of neurotensin and enteroglucagon release is mediated entirely through the guanine nucleotide-binding protein (Ni) or a similar toxin-sensitive, GTP-binding protein. SRIF-inhibited peptide release is mediated partially through a toxin-sensitive substrate, as evidenced by PT reversal of reduced cAMP levels. SRIF may also inhibit neurotensin and enteroglucagon release by a cAMP-independent pathway that is not coupled to Ni or a similar PT-sensitive, GTP-binding protein.
Neural regulation of gastrin and somatostatin secretion in rat gastric antral mucosa
Authors: Schubert M. L. , Makhlouf G. M.
Published: December 2017

A muscle-stripped mucosal sheet obtained from rat antrum was mounted in an Ussing chamber and used to examine the regulation of gastrin and somatostatin secretion by antral neurons. The neurons were activated pharmacologically with 1,1-dimethyl-4-phenylpiperazinium (DMPP) or electrically by field stimulation using aluminum foil electrodes layered over the edge of the mucosal sheet. Field stimulation caused an increase in gastrin and somatostatin secretion that was dependent on the strength of the stimulus (10-30 V). Field stimulation at 30 V, 10 Hz, 4 ms caused a sixfold increase in gastrin and a twofold increase in somatostatin secretion that were nearly abolished by tetrodotoxin (85-89% inhibition). Atropine partially inhibited the gastrin response (34 +/- 6%) but had no effect on the somatostatin response. DMPP caused lesser, though significant, increases in gastrin (166%) and somatostatin (83%) secretion that were nearly abolished by hexamethonium (84-91%) but were not significantly affected by atropine. The increase in somatostatin secretion caused by DMPP and field stimulation, as well as the resistance of gastrin and somatostatin secretion to atropine, was consistent with preferential activation of noncholinergic neurons at the stimulatory modalities used. The pattern and magnitude of gastrin and somatostatin response to pharmacological and electrical stimulation of antral neurons were similar to those previously observed in the vascularly perfused whole rat stomach.
Histamine increases phosphorylation of 27- and 40-kDa parietal cell proteins
Authors: Chew C. S. , Brown M. R.
Published: December 2017

Purified, hormonally responsive parietal cells from rabbit gastric mucosae were used as a model to study intracellular mechanisms controlling parietal cell HCl secretion. Using a high-resolution, two-dimensional electrophoretic technique, we demonstrate that histamine increases phosphorylation of two parietal cell proteins with approximate molecular weights of 27 and 40 kDa and respective pIs of 5.9 and 6.2. The increase in phosphorylation appears to be mediated via an adenosine 3',5'-cyclic monophosphate (cAMP)-dependent mechanism because cAMP analogues and forskolin stimulate phosphorylation of these proteins, whereas the cholinergic agonist, carbachol, which elevates parietal cell intracellular free calcium concentration but not cAMP content, and the calcium ionophore, ionomycin, do not. Both phosphoproteins are located in low-speed particulate fractions. The 40-kDa phosphoprotein was found in both enriched chief and parietal cells. This phosphoprotein may be cytoskeleton associated, since it is detected in a Triton-insoluble particulate fraction after prolonged exposure of parietal cells to Triton X-100. The 27-kDa phosphoprotein was detected in parietal but not in enriched chief cells and appeared to be localized in a low-speed fraction previously shown to contain increased H+-K+-ATPase activity after histamine stimulation. The location and rapid increase in phosphorylation of the 27-kDa phosphoprotein upon histamine stimulation make this protein an attractive candidate for future studies of intracellular regulation of parietal cell HCl secretion. The 40-kDa phosphoprotein may play a more general role in control of cytoskeletal activity, and perhaps, in morphological transformations associated with stimulus-secretion coupling.
Rat parietal cell function after prolonged inhibition of gastric acid secretion
Authors: Larsson H. , Carlsson E. , Ryberg B. , Fryklund J. , Wallmark B.
Published: December 2017

Female rats were treated orally for 3 mo with omeprazole (40 and 400 mumol/kg). Both doses caused total inhibition of gastric acid secretion and recovery was parallel to that of H+-K+-ATPase activity (30-50 and 60-80% inhibition 24 h after doses, respectively). The H+-K+-ATPase activity returned to control levels within 1 wk after the last dose. Plasma gastrin levels were dose-dependently increased during treatment but reversed to control levels within 9 days after the last dose. Parallel with a general increase in corpus mucosal mass, both pepsinogen and H+-K+-ATPase total content increased. However, their tissue concentrations did not differ from control values, suggesting that neither parietal nor chief cell density are changed by omeprazole treatment and also that their growth is parallel to the general hyperplasia. In contrast, the oxyntic mucosal histamine concentration was increased, indicating an increase in the enterochromaffin-like (ECL) cell density. Maximal capacity of the mucosa to secrete acid increased in parallel with the increase in mucosal mass and total H+-K+-ATPase content. However, basal acid secretion did not differ between treatment groups. Increased capacity slowly declined toward control levels over the 70-day recovery period after withdrawal of omeprazole. These results suggest that hypergastrinemia, induced in the rat by pharmacological inhibition of gastric acid secretion, causes a hyperplasia of oxyntic mucosal cells, ECL cells growing faster than the others. The hyperplastic mucosa has an increased capacity to produce acid and is functionally normal.
Structure-activity relationship studies on cholecystokinin: analogues with partial agonist activity
Authors: Galas M. C. , Lignon M. F. , Rodriguez M. , Mendre C. , Fulcrand P. , Laur J. , Martinez J.
Published: December 2017

In the present study, hepta- and octapeptide analogues of the C-terminal part of cholecystokinin, modified on the C-terminal phenylalanine residue, were synthesized. CCK analogues were prepared in which the peptide bond between aspartic acid and phenylalanine had or had not been modified and were lacking the C-terminal primary amide function. These CCK derivatives were able to cause full stimulation of amylase release from rat pancreatic acini but without a decrease in amylase release at supramaximal concentrations. There was a close relationship between the abilities of these derivatives to stimulate amylase release and their abilities to inhibit binding of 125I-BH-CCK-9 to CCK receptors on rat and guinea pig pancreatic acini. These CCK analogues were also able to recognize the guinea pig brain CCK receptors, some of them being particularly potent. The findings indicate that the aromatic ring of phenylalanine is important for the binding to brain and pancreatic CCK receptors, whereas the C-terminal primary amide function is not essential for the binding to pancreatic CCK receptors but is crucial for biological activity of rat pancreatic acini.
Role of blood flow in gastric acid secretion
Authors: Holm L. , Perry M. A.
Published: December 2017

The relationship between gastric acid secretion and blood flow has been investigated with a variety of different blood flow techniques including aminopyrine clearance, hydrogen gas clearance, intravital microscopy, laser-Doppler flowmetry, radioactive microspheres, and the elimination of inert gases. The most commonly used technique, aminopyrine clearance, predicts that increasing acid secretion is accompanied by a parallel increase in blood flow. However, the efficiency of clearance of aminopyrine is low in the nonsecreting stomach and increases as secretion rate increases. This precludes the use of aminopyrine clearance as a reliable measure of gastric mucosal blood flow at all but the highest steady-state level of acid secretion and casts doubt on the findings with this technique. Other methods for measuring blood flow indicate that there is no simple relationship between secretion and flow, with some studies finding that secretion and flow change in parallel and others finding that secretion varies quite independently of flow to the mucosa. One consistent finding is a strong correlation between stimulated acid secretion and gastric oxygen consumption. Both acid secretion and oxygen consumption fall if celiac blood flow is reduced below a critical value, which in the anesthetized dog stomach is approximately 30-40 ml.min-1.100 g-1. Driving blood flow above this value does not increase oxygen consumption and acid secretion, i.e., they reach a plateau. The shape of this relationship with its flow-dependent and flow-independent portions is used to explain the apparently contradictory findings in the literature regarding gastric acid secretion and blood flow.
Triton WR-1339-induced changes in serum lipids and biliary lipid secretion
Authors: Zeniya M. , Reuben A.
Published: December 2017

Biliary lipid secretion rates were measured in fed rats after an intravenous injection of Triton WR-1339 (TWR, 60 mg/100 g body wt), an agent that inhibits lipoprotein removal from the circulation. Serum triglyceride, phospholipid (PL), and cholesterol (CH) concentrations rose within 3 h of TWR to 45, 6.6, and 10 times control values, respectively, at 24-36 h. Serum lipids fell rapidly at 48 h and were normal by 72-96 h after TWR. TWR did not alter bile flow, hepatic bile acid transport, or biliary bile acid output. Within 0.5 h of TWR, biliary PL and CH outputs fell greater than 70%, and taurocholate-stimulated biliary PL secretion was markedly reduced. Biliary PL and CH secretion rates were approximately 30 and approximately 40% suppressed, respectively, 24 h after TWR, 160 and 330% elevated, respectively, at 48 h, and normally by 72 h, despite normal taurocholate-stimulated biliary PL secretion. Biliary beta-glucuronidase secretion (a lysosomal enzyme) was unchanged for 3 h after TWR but was increased at 24, 48, and 72 h, independent of biliary lipid secretion. Thus TWR acutely dissociates bile acid from lipid secretion without impairing bile acid transport or biliary lysosomal discharge. Late changes in biliary lipid secretion relate closely to TWR-induced change in serum lipid metabolism but not to stimulation of biliary lysosomal discharge.
Altered regulation of intestinal ion transport by enteric nerves in diabetic rats
Authors: Perdue M. H. , Davison J. S.
Published: December 2017

We compared ion transport parameters in isolated ileal mucosa from diabetic rats (8 wk after streptozotocin injection) and littermate controls under basal conditions and in response to electrical transmural stimulation (TS). Stripped ileal mucosa (submucosal plexus intact) was mounted in Ussing flux chambers modified to include stimulating electrodes on opposite sides of the tissue. Under basal conditions unidirectional fluxes of Na+ and Cl- were decreased across mucosa from diabetic rats compared with controls, whereas net fluxes were not significantly different. TS caused a tetrodotoxin (TTX)-sensitive transient increase in short-circuit current (Isc) that was significantly less in tissue from diabetic than control rats. The muscarinic cholinergic receptor antagonist, atropine, significantly reduced the Isc response to TS in ileum from control but not diabetic rats. In addition, the noncholinergic component of the response was smaller. The muscarinic agonist, Urecholine chloride (bethanechol chloride), caused an increase in Isc that was unaffected by pretreatment with TTX and was the same in tissue from control and diabetic rats. Our results suggest that the intestinal abnormalities that occur in diabetes may include a defect in the regulation of ion transport by enteric nerves resulting in an abnormal ability to respond to luminal and other stimuli.
Antropyloroduodenal motor responses to intraduodenal lipid infusion in healthy volunteers
Authors: Heddle R. , Dent J. , Read N. W. , Houghton L. A. , Toouli J. , Horowitz M. , Maddern G. J. , Downton J.
Published: December 2017

The delivery of lipid to the duodenum has been shown to slow gastric emptying and to increase the resistance to gastric outflow. To investigate mechanisms responsible for these effects, we have recorded antropyloroduodenal motility in nine healthy volunteers during alternate intraduodenal infusions of normal saline and triglyceride emulsion (Intralipid 10%). During the lipid infusions there were reproducible, major changes in the patterns of motility. Pressure waves, apparently isolated to the pylorus, usually started within 10 min of initiation of the lipid infusion. After 20-25 min of lipid infusion these waves occurred at median rates of 2.4 and 2.8/min (1st and 2nd lipid infusions, respectively); these rates were significantly greater (P less than 0.05) than the median rates (all less than or equal to 0.4/min) observed during the equivalent period of the succeeding saline infusions. During 10 of 22 lipid infusions, isolated pyloric pressure waves were associated with sustained pyloric tone. Infusion of lipid into the duodenum suppressed antral pressure waves in all subjects and initiated brief periods of regular duodenal contractions during 11 of 22 infusions. These studies have demonstrated alterations of antropyloroduodenal motor patterns in response to changes in the duodenal luminal content. The effects on antral and pyloric motility are probably of importance in the regulation of transpyloric flow by nutrients in the duodenal lumen.
Development of the muscarinic receptor in rabbit gastric smooth muscle
Authors: Tomomasa T. , Hyman P. E. , Hsu C. T. , Jing J. , Snape W. J.
Published: December 2017

We used binding of [N-methyl-3H]scopolamine ( [3H]-NMS) to tissue homogenates and isometric contraction of muscle strips to characterize perinatal changes in the muscarinic receptor on rabbit gastric smooth muscle. In homogenates from fetal (28 days of gestation), 1-, 3-, and 7-day, 4- and 11-wk-old rabbits, specific binding was saturable and temperature dependent, achieved equilibrium by 10 min at 30 degrees C, and was linearly related to tissue concentration. Specific binding was 80 +/- 2% of total binding at 0.2 nM [3H]NMS. The number of binding sites was 120,000 receptors/cell, maximal during the first week of life compared with the fetus or older animals. Affinity of [3H]NMS was highest in the first week of life (Kd = 345 +/- 24 pM, 1 day old). Age did not affect Hill coefficients or Ki values; secoverine and 4-diphenylacetoxy-N-methylpiperidine methiodide were 50-fold more potent than pirenzepine. In muscle strips, bethanechol stimulated dose-dependent atropine-inhibitable isometric contraction. The doses required for half-maximal contraction were similar in both age groups (5-6 microM), but maximal contraction was fivefold greater in weanlings compared with neonates. Increasing extracellular potassium concentration resulted in similar differences, suggesting that the differences were not receptor related. These results suggest that well-differentiated M2-muscarinic receptors are functional on rabbit gastric smooth muscle during the perinatal period.
Estrogen modulates ileal basolateral membrane lipid dynamics and Na+-K+-ATPase activity
Authors: Schwarz S. M. , Bostwick H. E. , Medow M. S.
Published: December 2017

The effects of ethinyl estradiol, a synthetic estrogen with cholestatic properties and a propensity to alter hepatocyte and ileal brush-border membrane fluidity, on lipid structure and Na+-K+-ATPase activity of rabbit small intestinal basolateral membranes were determined. Utilizing the fluorophores 1,6-diphenyl-1,3,5-hexatriene and DL-12-(9-anthroyl)stearic acid, increases in fluorescence anisotropy, the reciprocal of fluidity, were found in basolateral membranes and in membrane lipid liposomes isolated from ileum. Fluidity alterations were accompanied by a marked decrease in bilayer phospholipids (0.37 vs. 0.48 mumol/mg protein; P less than 0.01) and an increase in both the cholesterol-to-phospholipid molar ratio (0.85 vs 0.61; P less than 0.02) and membrane saturated fatty acid content. Estrogen-mediated physicochemical changes were associated with a significant reduction in ileal basolateral membrane Na+-K+-ATPase specific activity (100.0 vs. 185.8 nmol Pi.min-1.mg protein-1; P less than 0.02). Control values both for fluorescence anisotropy and for Na+-K+-ATPase specific activity were restored after in vitro membrane fluidization with benzyl alcohol. The data therefore indicate that ethinyl estradiol effects on basolateral membrane lipid dynamics are confined to the ileum and are associated with inhibition of Na+-K+-ATPase activity. These structural and functional changes appear to be related, in part, to specific modifications in the availability of phospholipid after estrogen treatment.
Thromboxane synthesis inhibitors and postprandial jejunal capillary exchange capacity
Authors: Mangino M. J. , Chou C. C.
Published: December 2017

The effects of thromboxane synthesis inhibitors (imidazole and U 63557A; Upjohn) and the cyclooxygenase inhibitor, mefenamic acid, on jejunal capillary filtration coefficients (Kfc) were determined in dogs before and during the presence of predigested food in the jejunal lumen. The jejunal Kfc increased significantly soon after the placement of a predigested test food containing all major constituents of diet. The Kfc remained elevated as long as the food was present in the lumen (15 min). Mefenamic acid (10 mg/kg iv) did not significantly alter resting jejunal Kfc or alter the food-induced increase in Kfc. Imidazole (5.0 mg/min ia) or U 63557A (5.0 mg/kg iv) per se significantly increased jejunal Kfc. Placement of digested food further increased the Kfc to levels significantly higher than those observed before administration of the two thromboxane synthase inhibitors. Production of thromboxane B2 by jejunal tissue was significantly reduced and 6-ketoprostaglandin F1 alpha (the stable hydrolysis product of prostacyclin) production was significantly increased after administration of U 63557A. Our study indicates that the relative production of endogenous thromboxanes and other prostanoids modulates jejunal capillary exchange capacity in the absence or presence of digested food in the jejunal lumen.
Lipoprotein lipase uptake by the liver: localization, turnover, and metabolic role
Authors: Vilaro S. , Llobera M. , Bengtsson-Olivecrona G. , Olivecrona T.
Published: December 2017

We have studied the binding and metabolism of 125I-labeled bovine lipoprotein lipase (LPL) by use of isolated, perfused rat livers. Our data suggest the presence of two types of binding sites, i.e., heparin-sensitive sites that bind primarily the catalytically active form of the lipase and are present at the endothelium in all blood vessels and heparin-insensitive sites that bind both active and inactive forms and are present only within the sinusoids. Forty minutes after uptake by the liver, approximately 50% of the LPL had lost its catalytic activity or been degraded. Three processes were evident: 1) colchicine-sensitive degradation to acid-soluble products, 2) partial proteolysis to fragments similar to those formed by limited digestion with trypsin or plasmin, and 3) a conformational change leading to loss of catalytic activity. Exogenous LPL bound in the liver caused a dramatic increase in the utilization of a perfused triacylglycerol emulsion (Intralipid), with rapid formation of free fatty acids and water-soluble metabolites. When the liver was flushed with heparin, it lost its ability to utilize the fat emulsion. Measurement of the hepatic extraction showed that rat livers take up 100-200 mU endogenous LPL per hour.
Lymphatic function in the liver after hepatic venous pressure elevation
Authors: Elk J. R. , Drake R. E. , Williams J. P. , Gabel J. C. , Laine G. A.
Published: December 2017

The liver lymphatic system plays an important role in removing excess fluid from the hepatic tissue. A complete analysis of the liver lymphatic system would be difficult. However, we used a simple circuit-analysis technique to represent the intrahepatic portion of the lymph system as a single pressure source (PL) pushing lymph through a single resistance (RL). Liver lymphatic vessels were cannulated in nine halothane-anesthetized dogs. The lymphatic vessel outflow pressure (PO) was varied by raising the outflow end of the cannula. Lymph flow from the cannula (QL) decreased linearly with PO, and we calculated RL as -delta PO/delta QL and PL as the extrapolated PO at which QL = 0. At base line, PL = 8.5 +/- 2.9 cmH2O, and RL = 0.05 +/- 0.03 cmH2O.min/microliter. After we increased inferior vena caval pressure from 5.8 +/- 2.7 to 15.2 +/- 2.5 cmH2O, PL increased significantly to 13.7 +/- 3.4 cmH2O, and RL decreased to 0.02 +/- 0.02 cmH2O.min/microliter (P less than 0.05). The results indicate that increases in QL occur because the effective pressure pushing lymph from the liver (PL) increases, and the effective resistance of the intrahepatic lymph vessels (RL) decreases.
Primary culture of PYY cells from canine colon
Authors: Aponte G. W. , Taylor I. L. , Soll A. H.
Published: December 2017

In the present study we report methods for the isolation and culture of colonic peptide YY (PYY) cells and have tested the effects of sodium oleate and other chemotransmitters on PYY release. Enzyme-dispersed canine colonic mucosa cells were separated by a Beckman elutriation rotor, and the enriched PYY fraction was cultured on type I collagen in full growth medium. After 42-48 h of culture PYY cells had selectively adhered to the collagen. Forty to 45% of the adherent cells contained PYY (5.5 +/- 0.5 pmol/24-mm well), and 10-15% of the cells contained glucagon-like immunoactivity (GL29-LI; 0.95 pmol/24-mm well). The effect of various secretagogues on PYY release from these cultures was monitored. Sodium oleate stimulated PYY release in a time-dependent fashion over a dose range from 10 microM to 10 mM. However, sodium oleate at a dose of 100 mM produced disproportionately large PYY release. During a 2-h incubation 5.1% of the total cell content of PYY was released in response to 0.1 mM sodium oleate compared with basal release of 1.1%. At doses less than 10 mM sodium oleate did not release GL29-LI, whereas concentrations of 10 and 100 mM resulted in marked GL29-LI release. These findings suggest that lower concentrations of sodium oleate selectively release PYY, whereas higher concentrations nonselectively induce peptide release probably by effects on membrane stability. We also found that bombesin, epinephrine, and forskolin, but not carbachol, produced time- and dose-dependent release of PYY from these cultures.
Effects of a high-protein, low-carbohydrate diet on degradation of sucrase-isomaltase in rat jejunoileum
Authors: Goda T. , Raul F. , Gosse F. , Koldovsky O.
Published: December 2017

During the degradation of intestinal sucrase-isomaltase by pancreatic proteinases, degradation of sucrase-active site precedes that of the isomaltase-active site in rats. In the present paper, we demonstrate that the extent of degradation of sucrase-isomaltase is altered by dietary manipulation in vivo. Adult rats were starved for 24 h and received either a standard diet (20 cal% protein, 55% carbohydrate) or an isocaloric high-protein, low-carbohydrate diet (70 cal% protein, 5% carbohydrate). Animals were killed 15 h after the refeeding. In rats fed a high-protein, low-carbohydrate diet, luminal trypsin activity was three times higher than controls, and sucrase activity in proximal ileum was significantly lower (P less than 0.001) than controls, whereas isomaltase activity was similar in both groups. In proximal jejunum, luminal trypsin activity was remarkably lower (P less than 0.01) than in proximal ileum in both groups; sucrase and isomaltase activity was similar in both groups. Crossed immunoelectrophoresis demonstrated that a degradation product of sucrase-isomaltase, i.e., isomaltase monomer, was present in a larger amount in rats fed a high-protein, low-carbohydrate diet. In rats with bypassed pancreatic ducts, the amount of this degradation product was decreased and effect of a high-protein, low-carbohydrate diet was abolished. Experiments with a sequential isolation of epithelial cells of proximal ileum revealed that sucrase activity was decreased along the entire height of the villus in animals fed a high-protein, low-carbohydrate diet.(ABSTRACT TRUNCATED AT 250 WORDS)
Functional somatostatin receptors on a rat pancreatic acinar cell line
Authors: Viguerie N. , Tahiri-Jouti N. , Esteve J. P. , Clerc P. , Logsdon C. , Svoboda M. , Susini C. , Vaysse N. , Ribet A.
Published: December 2017

Somatostatin receptors from a rat pancreatic acinar cell line, AR4-2J, were characterized biochemically, structurally, and functionally. Binding of 125I-[Tyr11]somatostatin to AR4-2J cells was saturable, exhibiting a single class of high-affinity binding sites (Kd = 0.55 +/- 0.06 nM) with a maximal binding capacity of 258 +/- 20 fmol/10(6) cells. Somatostatin receptor structure was analyzed by covalently cross-linking 125I-[Tyr11]somatostatin to its plasma membrane receptors. Gel electrophoresis and autoradiography of cross-linked proteins revealed a peptide (Mr 80,000) containing the somatostatin receptor. Somatostatin inhibited vasoactive intestinal peptide (VIP)-stimulated adenosine 3',5'-cyclic monophosphate (cAMP) formation in a dose-dependent manner. The concentration of somatostatin that caused half-maximal inhibition of cAMP formation (IC50 = 0.4 nM) was close to the receptor affinity for somatostatin. Pertussis toxin pretreatment of AR4-2J cells prevented somatostatin inhibition of VIP-stimulated cAMP formation as well as somatostatin binding. We conclude that AR4-2J cells exhibit functional somatostatin receptors that retain both specificity and affinity of the pancreatic acinar cell somatostatin receptors and act via the pertussis toxin-sensitive guanine nucleotide-binding protein Ni to inhibit adenylate cyclase.
Caerulein causes translocation of protein kinase C in rat acini without increasing cytosolic free Ca2+
Authors: Bruzzone R. , Regazzi R. , Wollheim C. B.
Published: December 2017

We investigated the relationships between changes in cytosolic free Ca2+ ([Ca2+]i) and amylase secretion in dispersed rat pancreatic acini. Although 10 pM caerulein did not raise [Ca2+]i, higher concentrations (1 nM) of the peptide elicited a prompt, marked, but transient (2-3 min) elevation of [Ca2+]i. Both concentrations of caerulein caused an almost identical release of amylase over a 30-min period. To investigate the mechanism(s) underlying Ca2+-independent secretion, we measured the effect of the secretagogue on protein kinase C activity and found that both caerulein concentrations caused a significant translocation of protein kinase C from the cytosolic to the microsomal fraction. Because 1 nM caerulein induced a greater enzyme secretion than 10 pM caerulein during the first 2-5 min of stimulation, we explored further the role of [Ca2+]i transients during the first minutes of secretion. Addition of ionomycin in the presence of 10 pM caerulein resulted in a rise in [Ca2+]i and enhanced secretion as a result of caerulein in a near additive fashion during the first 2 min of stimulation. Second, we pretreated acini for 5 min with 1 microM 12-O-tetradecanoylphorbol-13-acetate. This maneuver inhibited both caerulein-induced inositol trisphosphate formation and [Ca2+]i elevation. These findings were paralleled by a similar inhibition of caerulein-stimulated amylase release only during the first 5 min of secretion. These results indicate that 1) caerulein can stimulate amylase secretion independently of a concomitant [Ca2+]i rise, possibly by activation of protein kinase C, and 2) an elevation of [Ca2+]i serves as a trigger to enhance amylase release only during the initial phase of secretion.
Transport of guanidine in rabbit intestinal brush-border membrane vesicles
Authors: Miyamoto Y. , Ganapathy V. , Leibach F. H.
Published: December 2017

The characteristics of guanidine uptake were studied in brush-border membrane vesicles isolated from the rabbit proximal intestine. Guanidine uptake was manyfold greater in the presence of an outward-directed H+ gradient (intracellular pH = 5.5; extracellular pH = 7.2) than in the absence of a H+ gradient (intracellular and extracellular pH = 7.2). The time course of guanidine uptake exhibited an overshoot phenomenon in the presence of the H+ gradient, indicating occurrence of uphill transport. This H+ gradient-stimulated guanidine uptake was not due to an inside-negative H+-diffusion potential because carbonyl cyanide 4-trifluoromethoxyphenylhydrazone, a protonophore, failed to have any effect on guanidine uptake. Moreover, the transient uphill transport of guanidine was observed even in voltage-clamped membrane vesicles. However, under the conditions that effectively dissipated the H+ gradient, there was no active transport of guanidine. This H+ gradient-dependent transport mechanism for guanidine is distinct from the Na+-H+ exchanger, because amiloride did not inhibit guanidine uptake even at a concentration as high as 100 microM. These data provide evidence for the presence of a guanidine-H+ antiport system in the rabbit intestinal brush-border membrane. The presence of a carrier for guanidine in these membranes is further substantiated by the trans-stimulation of the uptake of radiolabeled guanidine by unlabeled guanidine and by the inhibition of guanidine uptake by imipramine under equilibrium exchange conditions.(ABSTRACT TRUNCATED AT 250 WORDS)
Carbamoylcholine and gastrin induce inositol lipid turnover in canine gastric parietal cells
Authors: Chiba T. , Fisher S. K. , Park J. , Seguin E. B. , Agranoff B. W. , Yamada T.
Published: December 2017

The potential role of inositol phospholipid turnover in mediating acid secretion was examined in a preparation enriched for isolated canine gastric parietal cells. The stimulatory effects of carbamoylcholine (carbachol) and gastrin on parietal cell uptake of [14C]aminopyrine were linked to dose- and time-dependent selective reduction in cellular phosphatidylinositol content, although the specific fatty acid composition of the phosphoinositides was not altered. Analysis of [3H]inositol phosphates accumulated in cells prelabeled with [3H]inositol revealed an increase in labeled inositol trisphosphate by 5 min of incubation with either carbachol or gastrin. Furthermore, after preincubation of parietal cells in medium containing [32P]orthophosphate, the two secretagogues elicited a time-dependent decrease in 32P labeling of phosphatidylinositol 4,5-bisphosphate and concomitant increase in labeling of phosphatidic acid. These data demonstrate that the acid secretagogue actions of carbachol and gastrin are correlated with turnover of cellular inositol phospholipids in a preparation consisting predominantly of parietal cells.
Glucose enhancement of transcellular calcium transport in the intestine
Authors: Carroll K. M. , Wood R. J. , Chang E. B. , Rosenberg I. H.
Published: December 2017

Glucose stimulates calcium transport in vitro in rat duodenal tissue and isolated enterocytes. Under short-circuited conditions, glucose increased mucosal to serosal calcium flux (JCa(m----s)) without altering serosal to mucosal calcium flux (JCa(s----m)) in the duodenum, the primary site of active calcium absorption in the rat small intestine. The half-maximal dose (ED50) of the glucose stimulatory effect was less than 1 mM, and an increase in JCa(m----s) of 80% over control was seen at a glucose concentration of 50 mM. Glucose did not increase calcium flux in the ileum where active calcium absorption is minimal. Glucose stimulated net calcium uptake by 35% in isolated duodenal enterocytes. Glucose did not alter calcium efflux from preloaded enterocytes suspended in calcium-free buffer. Glucose enhancement of net calcium uptake in enterocytes was not caused by altered cell membrane integrity or functional viability. The nonmetabolizable glucose analogue alpha-methylglucoside did not stimulate calcium transport. Our findings suggest that glucose can stimulate intestinal calcium absorption, at least partially, by enhancing transcellular calcium transport and that cellular glucose metabolism is necessary for stimulation of this route of calcium transport.
Influence of purinoceptors' agonists and antagonists on opossum internal anal sphincter
Authors: Rattan S. , Shah R.
Published: December 2017

Studies were performed in alpha-chloralose-anesthetized and pancuronium-treated opossums. Resting internal anal sphincter pressures (IASP) were monitored using low-compliant continuously perfused catheters. P1 and P2 purinoceptor agonists, adenosine and ATP, respectively, administered close intra-arterially, caused dose-dependent decreases in the IASP. The inhibitory effect of these agonists on the IASP was tetrodotoxin resistant. Rectal balloon distension (RD) (which mimics the rectoanal inhibitory reflex) caused volume-dependent IAS relaxation. Electrical stimulation of the sacral nerve (SNS) also produced frequency-dependent decreases in IASP. The inhibitory response to adenosine (P1 purinoceptor agonist), ATP (P2 purinoceptor agonist), RD, and SNS on the internal anal sphincter (IAS) was examined before and after 8-phenyltheophylline (P1 purinoceptor antagonist) and alpha,beta-methylene ATP (P2 purinoceptor antagonist that irreversibly binds and desensitizes P2 purinoceptor). P1 and P2 purinoceptor antagonists produced selective antagonism of the inhibitory responses on the IAS of their respective agonists only. Furthermore, high doses of adenosine and ATP produced desensitization and block of their own actions only. The purinoceptors' antagonists, and the desensitization of purinoceptors by high doses of adenosine and ATP, failed to modify the fall in IASP in response to RD or SNS. From these studies we conclude that distinct inhibitory P1 and P2 purinoceptors are present on the IAS smooth muscle. However, these inhibitory purinoceptors may not be responsible for the rectoanal reflex-mediated IAS relaxation.
Basal and PGE2-stimulated duodenal bicarbonate secretion in the rat in vivo
Authors: Heylings J. R. , Feldman M.
Published: December 2017

We studied basal and prostaglandin E2 (PGE2)-stimulated duodenal HCO3- transport in the rat in vivo both in the presence and absence of a concentration gradient for HCO3- from blood to lumen. Basal HCO3- transport was not reduced when the luminal solution was changed from one containing 0 mM HCO3- to one containing 22 mM HCO3- either at pH 9.0 or 7.5. Thus basal duodenal HCO3- transport in rats is independent of a blood-to-lumen HCO3- concentration gradient, which indicates an energy-dependent process with little passive flux of HCO3-. Luminal or intravenous administration of PGE2 significantly (P less than 0.01) increased HCO3- secretion into a HCO3(-)-free luminal solution but had no effect on HCO3- secretion into luminal solutions containing 22 mM HCO3-, either at pH 9.0 or 7.5. Therefore prostaglandins may act by increasing passive flux of HCO3- rather than by stimulating energy-dependent duodenal HCO3- transport.
Motility effects of intestinal anaphylaxis in the rat
Authors: Scott R. B. , Diamant S. C. , Gall D. G.
Published: December 2017

Intestinal motility was examined in an animal model of intestinal anaphylaxis. Hooded-Lister rats were sensitized (S) by intraperitoneal injection of 10 micrograms egg albumin (Ag) and compared with sham-sensitized controls (C). Seven days later three bipolar jejunal electrodes and a jejunostomy tube for motility recording and Ag administration were implanted. On day 14 intestinal myoelectric and motor activity were measured in fasted animals before and after intraluminal administration of either 10 mg egg albumin in 0.5 ml saline, 10 mg bovine serum albumin (BSA) in 0.5 ml saline, or placebo (P) challenge with 0.5 ml saline. Specific immunoglobulin E serum titers were greater than or equal to 1:64 in S animals, whereas C animals showed no response. None of the C animals challenged with P or Ag and none of the S animals challenged with P or BSA defecated after challenge, but all the S animals challenged with Ag developed diarrhea (P less than 0.001). In S animals challenged with Ag, the fasting motility pattern was disrupted, the migrating motor complex was abolished (P = 0.002), and the frequency of aborally propagating clustered contractions was increased (P less than 0.01). In this animal model an immune-mediated reaction to food protein was associated with diarrhea and altered intestinal myoelectric and motor activity.