bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of plasma membrane FABP and mitochondrial isoform of aspartate aminotransferase from rat liver
Authors: Stump D. D. , Zhou S. L. , Berk P. D.
Published: December 2017

A relationship between plasma membrane fatty acid binding protein (FABPpm), a putative membrane transporter for long-chain fatty acids, and the mitochondrial isoform of aspartate aminotransferase (m-AspAT) has been reported. Accordingly, we have compared the chemical and immunological properties of rat liver m-AspAT with those of rat liver FABPpm isolated by two procedures: 1) detergent solubilization of the membranes followed by purification via fatty acid affinity chromatography (FABP-1) or 2) salt extraction of the membranes and subsequent purification by high-performance liquid chromatography (HPLC; FABP-2). Comparison of the three protein preparations revealed no differences with respect to NH2-terminal amino acid sequence, amino acid composition, peptides from tryptic digests, AspAT enzymatic activity, isoelectric point, mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), retention on five different HPLC columns, and immunoprecipitation and immunoblotting of SDS-PAGE separated proteins with polyclonal antisera. Examination of the proteins by nondenaturing PAGE showed a consistent second band in FABP-1 and FABP-2 not always present in m-AspAT. However, whenever present, this band was immunoreactive with antibodies to both m-AspAT and FABP-1. Hence, FABP-1 and FABP-2 are indistinguishable from one another. They are also at least closely related, if not identical, to m-AspAT.
Effects of the inflammatory mediator prostaglandin D2 on submucosal neurons and secretion in guinea pig colon
Authors: Frieling T. , Rupprecht C. , Kroese A. B. , Schemann M.
Published: December 2017

Conventional flux chamber and intracellular recording methods were used to investigate the mode of action of prostaglandin D2 (PGD2) on ion transport in muscle-stripped segments of guinea pig colon and on colonic submucosal ganglion cells. Application of PGD2 resulted in a dose-dependent increase in short-circuit current that was reduced by serosal addition of bumetanide, tetrodotoxin, atropine, or piroxicam, but not hexamethonium. Application of PGD2 to submucosal neurons evoked a depolarization of the membrane potential that was associated with an enhanced spike discharge. In AH/type 2 neurons, postspike afterhyperpolarizations were reduced in amplitude and duration. The depolarizing responses to PGD2 were not affected by tetrodotoxin, indicative of a direct effect of PGD2 on the impaled neurons. Whereas fast excitatory postsynaptic potentials (EPSPs) were not affected by PGD2, slow EPSPs were reduced by a presynaptic effect, indicating presynaptic suppression of noncholinergic neurotransmitter release. The study demonstrates that PGD2 acts as a neuromodulator to evoke nerve-mediated chloride secretion, predominantly through activation of cholinergic submucosal neurons. The results further indicate that PGD2 released from lamina propria immune cells during antigenic stimulation may influence mucosal function by altering electrical behavior of submucosal neurons.
Polymorphonuclear leukocyte infiltration into gastric mucosa after ischemia-reperfusion
Authors: Andrews F. J. , Malcontenti-Wilson C. , O'Brien P. E.
Published: December 2017

Polymorphonuclear leukocytes (PMN) have been implicated in the pathogenesis of ischemia-reperfusion injury. The objective of this study was to investigate PMN infiltration and distribution within the gastric mucosa of rats subjected to 30 min gastric ischemia followed by reperfusion. Sections of mucosa were stained for PMN using an immunoperoxidase technique, and injury was assessed by quantitative histology. In control animals, there were 4 +/- 2 PMN/mm2 in the superficial and 9 +/- 4 PMN/mm2 in the deep mucosa. This increased significantly to 67 +/- 9 PMN/mm2 (P < 0.05) and 160 +/- 53 PMN/mm2 (P < 0.01) respectively at 15 min of reperfusion. The percentage of these PMN which were extravasated was 83 +/- 4% in the superficial mucosa and 82 +/- 4% in the deep mucosa (P < 0.001 compared with control levels of 0% in superficial and 10% in deep mucosa). Significant PMN infiltration occurred before full expression of mucosal injury and treatment of rats with anti-PMN antisera blocked reperfusion injury (treated 10.7 +/- 1.4% mucosa damaged, controls 33.5 +/- 2.4%; P < 0.001). Treatment with allopurinol (100 mg/kg) significantly reduced the number of infiltrating PMN (superficial 7 +/- 1/mm2, deep 16 +/- 2/mm2; P < 0.01) and the percentage of extravasating PMN (superficial 40 +/- 10%, deep 30 +/- 15%; P < 0.01) while also significantly reducing tissue injury (21.9 +/- 1.9% mucosa damaged, P < 0.01 compared with controls). We conclude that the immunoperoxidase staining provides a simple means of identifying PMN in histological sections. Furthermore, our results support a role for PMN in gastric ischemia-reperfusion injury.
Effects of prostaglandins on gastrin release from canine antral mucosal cells in primary culture
Authors: Schepp W. , Chan C. B. , Giraud A. S. , Avedian D. , Chen M. C. , Chew P. , Walsh J. H. , Soll A. H.
Published: December 2017

Evidence in vivo indicates that endogenous and exogenous prostaglandins can alter gastrin secretion. We have used primary cultures containing canine antral G-cells to study the cellular actions of prostaglandins on gastrin secretion, comparing the effects of prostaglandin E2 (PGE2) and its synthetic analogue enprostil. Enprostil (10(-10)-10(-6) M) inhibited gastrin secretion in response to bombesin, carbachol, and forskolin, the latter a receptor-independent activator of adenylate cyclase. This inhibition by enprostil was reversed by treatment with pertussis toxin (200 ng/ml, 8 h). However, enprostil did not inhibit the postreceptor stimuli 8-bromoadenosine 3',5'-cyclic monophosphate (10(-3) M), calcium ionophore A-23187 (10(-7) M), or 4 beta-phorbol 12-myristate 13-acetate (10(-8) M). In contrast, whereas PGE2 inhibited forskolin-stimulated gastrin release, PGE2 did not inhibit the response to carbachol or bombesin in control cultures. However, in pertussis toxin-treated cultures, PGE2 inhibition was reversed and, in contrast, the responses to bombesin, carbachol, and possibly forskolin were augmented. Indomethacin at a dose of 10(-5) M did not alter basal or bombesin-stimulated gastrin secretion. However, the somatostatin antibody CURE-S6 enhanced the response to forskolin and enhanced inhibition by PGE2, suggesting that endogenous somatostatin produced an inhibitory tone in these cultures and excluding the possibility that PGE2 acted via release of endogenous somatostatin. Our data suggest that in cultured antral cells gastrin release is regulated by inhibitory and stimulatory prostaglandin mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)
Evaluation of human postprandial antral motor function using ultrasound
Authors: Ahluwalia N. K. , Thompson D. G. , Mamtora H. , Troncon L. , Hindle J. , Hollis S.
Published: December 2017

We used high-resolution ultrasound to characterize postprandial antral excursion characteristics in 15 healthy volunteers. Antral circumference was repeatedly measured in the sagittal plane anterior to the abdominal aorta, at 15-min intervals following ingestion of a standard meal. At each recording period, the maximal and minimal antral circumference was noted. In seven volunteers, antral circumference was studied continuously for 5 min at each 15-min recording period to assess the frequency and regularity of antral excursions. Antral circumference increased immediately after meal ingestion from preprandial values of 64 +/- 2.6 (means +/- SE) to 138 +/- 3.1 mm and then gradually decreased with meal emptying to 72 +/- 2.6 mm at 90 min. The continuous recordings of antral images revealed that antral circumference transiently decreased at regular intervals; the interval between successive excursions was 20.1 +/- 0.2 s. There was no change in the excursion interval with emptying. The excursion amplitude decreased from 33 +/- 3.9 mm immediately post-meal ingestion to 13 +/- 1.5 mm at 90 min (r2 = 0.62 +/- 0.07). The ratio between the excursion amplitude and circumference of the relaxed antrum changed only slightly with time (slope = 0.0009 +/- 0.00003). Comparison of repeat studies in 10 volunteers showed a low intra-individual variation for postprandial antral circumference and excursion amplitude.
Beneficial effects of local or systemic lidocaine in experimental colitis
Authors: McCafferty D. M. , Sharkey K. A. , Wallace J. L.
Published: December 2017

Neuropeptides liberated from enteric neurons have been suggested to contribute to the inflammatory process in colitis. Based in part on this evidence, lidocaine enemas have recently been suggested as a treatment for ulcerative colitis, but their effects have yet to be fully characterized. We have assessed the effects of lidocaine on the development and maintenance of colitis induced by trinitrobenzenesulfonic acid (TNBS) in rats. In the initial experiments, rats were given lidocaine [5-100 mg/kg intrarectally] 30 min before TNBS administration, and the severity of colitis was assessed 6-24 h later. In subsequent experiments, rats received lidocaine intrarectally or subcutaneously 30 min before TNBS administration and once daily for 7 days. The severity of colitis was assessed by blind macroscopic scoring, measurement of myeloperoxidase activity, and histological evaluation. Lidocaine dose dependently reduced the severity of colitis and the infiltration of granulocytes at 24 h and 7 days post-TNBS. At the latter time point, lidocaine was also found to improve the histological appearance of the tissue and significantly reduce mucosal mast cell hyperplasia. Beneficial effects were also evident when intrarectal lidocaine treatment was initiated after induction of colitis or when lidocaine was given subcutaneously. Granulocyte infiltration into the colon could also be attenuated by a substance P receptor antagonist. These results suggest that lidocaine can exert anti-inflammatory effects in colitis, perhaps by virtue of its effects on intrinsic and/or extrinsic nerves.
Regulation of mucin secretion in T84 adenocarcinoma cells by forskolin: relationship to Ca2+ and PKC
Authors: Forstner G. , Zhang Y. , McCool D. , Forstner J.
Published: December 2017

The relationship between the adenosine 3',5'-cyclic monophosphate-mediated protein kinase A (PKA)-dependent stimulatory pathway for mucin secretion and Ca(2+)-mediated and protein kinase C (PKC)-mediated secretion was studied in T84 cells, using the postreceptor secretagogues forskolin, A-23187, and phorbol 12-myristate 13-acetate (PMA), the protein kinase inhibitors staurosporine and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), high- and low-Ca2+ media, and the Ca2+ chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). Staurosporine (10(-5) M) inhibited both PMA and forskolin at their maximally effective concentrations, whereas H-7 (5 x 10(-5) M) inhibited only PMA. Stimulation of mucin secretion by forskolin (5 x 10(-5) M) was not significantly affected by the reduction of medium Ca2+ to 47 and 129 nM, equivalent to published values for intracellular Ca2+ concentration ([Ca2+]i). Stimulation by forskolin was reduced by preloading cells with BAPTA, but to a much smaller extent than Ca(2+)-dependent stimulation by A-23187. A-23187-mediated mucin secretion from BAPTA-loaded cells was augmented by high doses of forskolin. Similar concentrations of forskolin had no effect on A-23187-stimulated secretion in calcium-replete cells. Our results indicate that forskolin does not stimulate mucin secretion by increasing Ca2+ entry or releasing Ca2+ from intracellular stores. Forskolin can stimulate mucin secretion in a Ca(2+)-independent manner but is apparently inhibited by high levels of intracellular Ca2+ induced by Ca2+ ionophores in 1.0 mM Ca2+ media.(ABSTRACT TRUNCATED AT 250 WORDS)
Brush-border tyrosine phosphorylation stimulates ileal neutral NaCl absorption and brush-border Na(+)-H+ exchange
Authors: Donowitz M. , Montgomery J. L. , Walker M. S. , Cohen M. E.
Published: December 2017

The drug genistein, a tyrosine (Tyr) kinase inhibitor, was used to define a role for Tyr phosphorylation in regulation of basal and stimulated neutral NaCl absorption in rabbit ileum. Brush-border vesicles contain Tyr-phosphorylated peptides. Genistein freeze-thawed into the vesicles caused a concentration-dependent inhibition of at least three peptides with M(r) 111,000, 83,000, and 80,000. Studied with the Ussing chamber-voltage clamp technique, genistein added to the ileal mucosal surface inhibited neutral NaCl absorption. Direct addition of genistein to brush-border vesicles made from ileal villus cells inhibited brush-border Na(+)-H+ exchange but not D-glucose-stimulated Na+ uptake. These effects were not duplicated by genistin, a drug with similar structure to genistein but lacking Tyr kinase inhibiting properties. Serosal but not mucosal epidermal growth factor (EGF) stimulated NaCl absorption. Mucosal genistein but not genistin also altered second-messenger regulation of neutral NaCl absorption, inhibiting the effect of Ca2+ ionophore A-23187 and of serosal EGF but not affecting the transport changes caused by 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP). In contrast, the Cl secretory effects indicated by the increase in short-circuit current for all three agents, A-23187, EGF, and 8-BrcAMP, were inhibited by mucosal genistein. These results suggest that 1) a Tyr kinase is involved in basally stimulating ileal neutral NaCl absorption and brush-border Na(+)-H+ exchange; 2) EGF stimulates NaCl absorption by an effect exerted from the serosal surface, but the effect also involves a brush-border Tyr kinase; 3) brush-border Tyr kinase is involved in the ability of Ca2+ ionophore A-23187 to inhibit neutral NaCl absorption but is not involved in the transport effects of cAMP. This study suggests that Tyr kinase(s) acting over short time periods is involved in stimulation of neutral NaCl absorption and brush-border Na(+)-H+ exchange and also in Ca(2+)-induced inhibition of NaCl absorption. These studies represent the first example of a brush-border Tyr kinase being involved in short-term signal transduction in intestinal epithelial cells.
Secretin activates Cl- channels in bile duct epithelial cells through a cAMP-dependent mechanism
Authors: McGill J. M. , Basavappa S. , Gettys T. W. , Fitz J. G.
Published: December 2017

Using patch-clamp recording techniques, we assessed the effects of secretin on membrane ion channel activity in isolated rat bile duct epithelial cells. In the whole cell configuration, secretin activated an inward membrane current at -40 mV in 6 of 13 cells, and increased current density from 17 +/- 8 to 98 +/- 33 pA/pF. Secretin-stimulated currents reversed near the equilibrium potential for Cl- and exhibited a linear current-voltage relationship. In the cell-attached configuration, secretin activated low-conductance channels in 73% (11 of 15) of patches. Similar channels were activated by forskolin, suggesting that adenosine 3',5'-cyclic monophosphate (cAMP) is involved as a second messenger. At the resting membrane potential, channels carried inward membrane current and had a slope conductance of 10 +/- 1 pS. In excised patches, addition of purified catalytic subunit of cAMP-dependent protein kinase (protein kinase A) to the cytoplasmic surface activated channels in four of six attempts. With equal Cl- concentrations in bath and pipette, channels had a linear slope conductance of 13 +/- 2 pS and currents reversed near 0 mV. Partial substitution of pipette Cl- with gluconate caused a shift in reversal potential in the direction anticipated for a Cl(-)-selective channel (gluconate to Cl- permeability ratio of 0.21 +/- 0.05, n = 4). Thus in bile duct epithelial cells, exposure to secretin activates low-conductance, Cl(-)-selective channels, probably through a cAMP-dependent mechanism. This likely contributes to secretin-dependent choleresis.
Upregulation of Escherichia coli heat-stable enterotoxin receptor in regenerating rat liver
Authors: Laney D. W. , Bezerra J. A. , Kosiba J. L. , Degen S. J. , Cohen M. B.
Published: December 2017

Guanylate cyclase C (GC-C) is a transmembrane protein that serves as a receptor for the recently characterized endogenous ligand guanylin and for Escherichia coli heat-stable toxin (STa). Binding of either guanylin or STa to intestinal GC-C results in net chloride secretion. Although GC-C is expressed in the rat intestine throughout life, its expression in the rat liver has previously been shown to occur only during the perinatal period. As a step toward elucidating the role of this receptor in the liver, we tested the hypothesis that GC-C mRNA expression could be induced in the adult rat liver following 1) partial hepatectomy, a stimulus for hepatocyte proliferation; 2) intraperitoneal carbon tetrachloride injection, a model of hepatocyte regeneration in the presence of inflammatory changes; and 3) subcutaneous turpentine injection, which generates an acute phase response without hepatocyte proliferation. We demonstrated expression of GC-C mRNA in the regenerating rat liver following either partial hepatectomy or CCl4-induced hepatic necrosis. We have also shown that GC-C mRNA expression occurred in association with an acute phase reaction. Coordinate with the expression of GC-C mRNA, there was upregulation of radiolabeled STa binding to liver plasma membranes prepared from turpentine-treated rats. Maximal expression of GC-C occurred in preparations enriched for the canalicular domain. Although the function of GC-C in the liver is unknown, localization to the canalicular domain would be consistent with a role for GC-C in hepatic chloride secretion, especially in the perinatal liver and during hepatocyte regeneration.
Rat pancreatic lipase and two related proteins: enzymatic properties and mRNA expression during development
Authors: Payne R. M. , Sims H. F. , Jennens M. L. , Lowe M. E.
Published: December 2017

We report the cDNA sequences of rat colipase, rat pancreatic lipase (rPL), and a rat pancreatic lipase-related protein (rPLRP). Comparison to the human PLRP cDNA suggests that the isolated clone encodes rPLRP-2. Both cDNA and a third cDNA encoding rPLRP-1 are secreted from Sf9 cells infected with recombinant baculovirus. rPL and rPLRP-2 hydrolyze triolein, 8.0 and 4.4 mumol.min-1.microgram-1, respectively. They are inhibited by bile salts, and activity is restored by (pro)colipase. PLRP-1 has barely detectable activity against triolein, even with (pro)colipase present. The pattern of mRNA expression during development in the rat reveals that all mRNA are low in the fetal rat pancreas. Both PLRP mRNA rise just before birth to a maximum 12 h after birth. They fall to low levels in the adult. In contrast, the PL mRNA is low at birth and rises rapidly during the suckling-weanling transition. In conclusion, the rat has at least three genes encoding different lipases, and these related genes have separate regulatory controls.
Nitric oxide and prostaglandins interact to prevent hepatic damage during murine endotoxemia
Authors: Harbrecht B. G. , Stadler J. , Demetris A. J. , Simmons R. L. , Billiar T. R.
Published: December 2017

Nitric oxide (NO) and prostaglandins (PG) both possess the ability to induce vasodilatation and prevent the aggregation of platelets. The synthesis of these substances is increased following in vivo lipopolysaccharide (LPS) infusion, but their function during sepsis is incompletely understood. We studied the role of NO and PG in a murine model of chronic hepatic inflammation (Corynebacterium parvum injection), which is known to progress to sudden hepatic necrosis after LPS injection. NO synthesis, which is induced in hepatocytes by C. parvum treatment and in nonparenchymal cells by LPS treatment, was inhibited using NG-monomethyl-L-arginine (L-NMMA). High-dose aspirin (ASA) was used to block PG synthesis. Treatment with L-NMMA or ASA alone, in the absence of LPS, resulted in no increase in hepatic injury. C. parvum-treated mice that received both L-NMMA and ASA without LPS developed marked hepatic damage as reflected by increased hepatocellular enzyme release (aspartate aminotransferase and L-ornithine carbamoyl-transferase). Marked hepatic damage was seen after LPS administration, and ASA pretreatment alone had no effect on the LPS-induced hepatic injury, whereas L-NMMA markedly increased the hepatic damage. The combination of L-NMMA and ASA after LPS resulted in the greatest hepatocellular enzyme release, characterized histologically by intravascular thrombosis with diffuse infarction and necrosis. Simultaneous treatment with either PGI2 or L-arginine partially prevented this injury. These data demonstrate that NO and PG function synergistically to maintain hepatocellular integrity; thus increased synthesis of these mediators protects the liver from the pathophysiological effects of LPS in this model.
Mechanism underlying tumor necrosis factor-alpha suppression of norepinephrine release from rat myenteric plexus
Authors: Hurst S. M. , Collins S. M.
Published: December 2017

We studied the effect of tumor necrosis factor-alpha (TNF-alpha) on the release of [3H]norepinephrine ([3H]NE) from longitudinal muscle-myenteric plexus preparations of rat jejunum. TNF-alpha had no immediate effect on [3H]NE release. Preincubation of the tissue with TNF-alpha caused a suppression of [3H]NE release stimulated by KCl or electrical field stimulation. The action of TNF-alpha was time and concentration dependent (0.1-50 ng/ml) and was not due to endotoxin contamination. The effect of TNF-alpha was biphasic, occurring after 30 min and again after 120 min of preincubation. The early component was independent of protein synthesis but was inhibited by piroxicam or indomethacin, indicating the involvement of cyclooxygenase metabolites. The late component was dependent on protein synthesis, was blocked by an interleukin-1 receptor antagonist, and was inhibited by piroxicam or indomethacin. These results indicate that TNF-alpha suppresses NE release by two mechanisms, one of which is due to the synthesis and release of interleukin-1, each involving arachidonic acid metabolites.
Cholinergic influence on duodenal mucosal bicarbonate secretion in the anesthetized rat
Authors: Safsten B. , Jedstedt G. , Flemstrom G.
Published: December 2017

Bicarbonate secretion by duodenum distal to the Brunner's glands area was titrated in situ in rats anesthetized with thiobarbiturate. The unselective muscarinic antagonist atropine (0.4 mg/kg) inhibited secretion stimulated by bethanechol (15 micrograms.kg-1.h-1) but not that stimulated by carbachol (15 micrograms.kg-1.h-1). The nicotinic antagonist hexamethonium (10 mg.kg-1.h-1), however, abolished the latter response. The muscarinic M1-selective antagonists pirenzepine and telenzepine (0.025, 0.25 and 2.5 mg/kg) did not decrease but caused a dose-dependent rise in duodenal mucosal HCO3- secretion, an effect abolished by cervical vagotomy or infusion of the alpha-adrenoceptor antagonist phentolamine (0.1 mg.kg-1.h-1). Phentolamine alone caused a sustained increase in secretion. McN-A-343 (0.025, 0.25, and 2.5 mg/kg), an M1-selective agonist and ganglionic stimulator, increased the HCO3- secretion; this effect was not prevented by vagotomy but was attenuated by pirenzepine. Intracerebroventricular infusion of pirenzepine and telenzepine did not cause any changes in secretion. These findings suggest that peripheral muscarinic M1 and nicotinic receptors mediate cholinergic stimulation of duodenal mucosal HCO3- secretion. Pirenzepine and telenzepine may act stimulatory by antagonizing muscarinic M1-transmission in peripheral sympathetic ganglia, thus decreasing postsynaptic adrenergic inhibition.
Integrative immunophysiology in the intestinal mucosa
Authors: Perdue M. H. , McKay D. M.
Published: December 2017

Over the past ten years, it has become evident that intestinal epithelial functions such as ion secretion are a host defense response to the presence of antigens, microbes, and other noxious substances in the gut lumen. Such responses are mediated by the activation of immune cells in the mucosa causing release of chemical mediators that act directly or indirectly on the epithelium. Frequently, immune cell products stimulate enteric nerves resulting in amplification. Thus immune cells and nerves form interactive units that can recognize various stimuli both specifically and nonspecifically and initiate mechanisms to eliminate offending material. Here, we review the current state of knowledge regarding immune regulation of epithelial physiology with particular emphasis on the ability of immune cells and their products (biogenic amines, cytokines, arachidonic acid metabolites, oxidants) to alter electrolyte transport. The mast cell will be highlighted in this scheme as this cell has been, and continues to be, the focus of extensive research efforts. However, recently it has become apparent that cells such as lymphocytes, macrophages, and polymorphonuclear leukocytes also play important roles in immunophysiology. The effect of immune cell activation on epithelial functions other than transport, such as permeability, proliferation, and antigen presentation, will be described where appropriate. Finally, we will present evidence that the enterocyte can express an "activated" phenotype and thus participate directly in mucosal immune responses.
Mechanism of action of insulin on pancreatic exocrine secretion in perfused rat pancreas
Authors: Lee K. Y. , Lee Y. L. , Kim C. D. , Chang T. M. , Chey W. Y.
Published: December 2017

In conscious rats, we have previously shown that immunoneutralization of circulating insulin with a rabbit anti-insulin serum abolished the pancreatic exocrine secretion stimulated by a meal or a combination of exogenous secretin and cholecystokinin octapeptide (CCK-8). To investigate the mechanism of endogenous insulin action on the exocrine pancreas, isolated rat pancreata were perfused with intra-arterial infusion of Krebs-Henseleit solution (37 degrees C) at 1.2 ml/min, whereas both pancreatic juice and portal venous effluent were collected separately in 15-min samples. Simultaneous intra-arterial infusion of secretin and CCK-8 in doses of 0.75 and 4.2 pmol/h; respectively, significantly increased volume, bicarbonate, and protein output in 7 rat pancreata (P < 0.01). When a rabbit anti-insulin serum was administered intra-arterially (0.1-ml bolus followed by 0.1 ml for 10 min), pancreatic secretion of volume, bicarbonate, and protein output was profoundly suppressed (n = 7, P < 0.01), whereas a normal rabbit serum failed to influence pancreatic secretion. The decrease in pancreatic secretion by the antiserum coincided with a significant increase in somatostatin in portal venous effluent from 1.4 +/- 0.2 to 4.1 +/- 0.8 pM (n = 6, P < 0.05). The combined administration of a rabbit antisomatostatin serum (0.4 ml) and the anti-insulin serum partially reversed the effect of the anti-insulin serum alone. Thus the pancreatic secretion was significantly greater than that achieved by the anti-insulin serum alone (P < 0.05). These observations strongly suggest that the action of insulin on exocrine pancreas is mediated by its local or paracrine action.(ABSTRACT TRUNCATED AT 250 WORDS)
Intestinal myoelectric response in two different models of acute enteric inflammation
Authors: Sjogren R. W. , Colleton C. , Shea-Donohue T.
Published: December 2017

The inflammatory responses to intraluminal administration of the cytotoxic lectin ricin or the hapten 2,4,6-trinitrobenzenesulfonic acid in 25% ethanol were compared. Myoelectric activity was recorded from in vivo ileal loops in rabbits after administration of either agent. At the end of a 6-h study the loops were removed and prepared for histological evaluation and for ex vivo generation of leukotriene (LT) C4 and prostaglandin (PG) E2. Both agents induced significant increases in mucosal damage, polymorphonuclear leukocyte infiltration, and edema. These changes tended to be patchy in the hapten model and confluent in the ricin model. Spike burst activity and mucosal LTC4 and PGE2 generation were also enhanced significantly. Intraluminal administration of the specific LTD4 antagonist Wy-48252 1 h before either inflammagen, significantly inhibited the myoelectric response without affecting tissue morphology or eicosanoid synthesis. These data demonstrate that there is a stereotypic spike burst response to acute inflammation, which is mediated in part by LTD4.
Role of a lymphatic system in glucose absorption and the accompanying microvascular hyperemia
Authors: Steenbergen J. M. , Lash J. M. , Bohlen H. G.
Published: December 2017

In this study we evaluated the importance of a functional intestinal lymphatic system on changes in arteriolar and venular blood oxygen content, vasodilation, and elevation of venous blood osmolarity during glucose absorption. Glucose absorption was associated with a doubling of the arteriovenous oxygen difference [(A-V)O2], a 50 mosM increase in venous blood osmolarity, and 17% dilation of the intermediate-diameter arterioles. After the lymph vessels were mechanically blocked with mineral oil, glucose absorption again doubled the (A-V)O2, indicating that glucose was absorbed without a functional lymphatic system. Furthermore, venous blood osmolarity and arteriolar diameter increased similarly with and without a functional lymphatic system. This study indicates that even though the lymphatic system likely facilitates distribution of hypertonic material in the bowel wall during absorption, blockade of the lymphatics did not appreciably hinder vasodilation, glucose absorption, changes in intravascular oxygen content, or the elevation of tissue hyperosmolarity, as judged by the tonicity of the venular blood. Therefore, passage of materials absorbed or released in the mucosa to the submucosa through venular blood flow may be very important to the mechanism of absorptive hyperemia.
Tumor necrosis factor-alpha induces c-jun during the regenerative response to liver injury
Authors: Diehl A. M. , Yin M. , Fleckenstein J. , Yang S. Q. , Lin H. Z. , Brenner D. A. , Westwick J. , Bagby G. , Nelson S.
Published: December 2017

After liver injury, remaining hepatocytes proliferate to regenerate the liver. Although the precise mechanisms that initiate and localize regeneration are unknown, local induction of c-jun is a critical, early step in the response. Treatment of rats with antibodies to tumor necrosis factor-alpha (TNF-alpha), a mediator of liver injury, inhibits regenerative induction of jun nuclear kinase activity and nuclear c-jun expression and alters the DNA binding activity of the c-jun transcription factor, AP-1, in liver. Pretreatment with anti-TNF antibodies does not affect pulmonary or renal c-jun expression or AP-1 binding activity post-partial hepatectomy. In primary hepatocyte cultures, TNF-alpha directly promotes the proliferative actions of mitogens, supporting in vivo evidence that it sensitizes hepatocytes to mitogens. Thus local release of TNF may act in a paracrine fashion to initiate regeneration in the injured liver by promoting induction of critical growth-related genes, such as c-jun.
Secretagogue regulation of pancreatic acinar cell protein phosphorylation shown by large-scale 2D-PAGE
Authors: Wishart M. J. , Groblewski G. , Goke B. J. , Wagner A. C. , Williams J. A.
Published: December 2017

High-resolution large-scale two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) combined with computer-assisted image analysis was used to construct a database of secretagogue/second messenger-induced phosphoprotein modifications in intact rat pancreatic acinar cells. Isolated acini were labeled with 32Pi, exposed to hormones and other test agents, and subjected to large-scale 2D-PAGE and autoradiography. This procedure resolved 500 phosphoproteins in pancreatic acinar whole cell lysates, approximately 90% of which were localized in the soluble fraction of centrifuged samples. Soluble proteins were further characterized as to heat and acid stability. Cholecystokinin (CCK), carbachol, and bombesin altered the phosphorylation state of about 27 proteins with both increases and decreases observed. Subsets of proteins were phosphorylated in response to phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA), calcium ionophore A-23187, and adenosine 3',5'-cyclic monophosphate (cAMP) analogue 8-bromo-cAMP. One of these proteins was identified as the myristoylated, alanine-rich, C-kinase substrate (MARCKS) protein by immunoprecipitation. The time course and dose response of phosphorylation changes due to CCK showed considerable variation between proteins, although a temporal hierarchy of phosphorylation events was clearly exhibited. Particularly striking was the rapid dephosphorylation within 30 s of a 19-kDa soluble protein to a minimum of 20 +/- 1% of control. Increased phosphorylation of the MARCKS and other TPA-regulated proteins suggests that CCK, carbachol, bombesin, and the CCK partial agonist, JMV-180, all activate protein kinase C in intact acini.
Intestinal anaphylaxis: radiation-induced suppression
Authors: Harari Y. , Kester D. , Travis E. , Wallace J. , Castro G.
Published: December 2017

The gastrointestinal tract is highly sensitive to ionizing radiation. Some of the most radiosensitive cells in this system are mast cells and epithelium. This article describes experiments that test the hypothesis that irradiation suppresses mucosal immune responses in which mast cells and epithelium are involved. The hypothesis was tested by examining the impact of ionizing radiation on anaphylactically mediated Cl- secretion in jejunum of rats sensitized by Trichinella spiralis infection and challenged with antigen derived from the parasite. Antigen-induced Cl- secretion was measured electrophysiologically in vitro. Rats were immunized by inoculation with 3 x 10(3) T. spiralis larvae and, 30-50 days later, exposed to total abdominal irradiation from a cobalt-60 gamma source. Doses were 5, 7, and 9 Gy. At 1, 3, 5, 7, 14, and 21 days postirradiation (DPI), jejunal segments were assessed for immune responsiveness. (Duration of suppression to antigenic challenge was directly related to radiation dose). Recovery of response to antigenic challenge after irradiation with 5 Gy was complete by 5 DPI. At 7 Gy, responsiveness was totally suppressed from 1 to 5 DPI, was partially expressed from 5 to 14 DPI, and was completely restored by 21 DPI. A dose of 9 Gy completely suppressed immune responsiveness throughout the 21-day period. Full responsiveness of jejunum to exogenous Cl- secretagogues at 1-5 DPI indicates that the immunosuppressive effect of radiation was not due to a breakdown in the secretory process at the epithelium level.(ABSTRACT TRUNCATED AT 250 WORDS)
Glutamine-enriched enteral diet increases splanchnic blood flow in the rat
Authors: Houdijk A. P. , Van Leeuwen P. A. , Boermeester M. A. , Van Lambalgen T. , Teerlink T. , Flinkerbusch E. L. , Sauerwein H. P. , Wesdorp R. I.
Published: December 2017

The hemodynamic consequences of glutamine (Gln)-enriched nutrition have not been investigated. This study investigates the effects of a Gln-enriched enteral diet on organ blood flows and systemic hemodynamics. Male Fischer 344 rats (n = 24) were randomized to a group that received a 12.5% (wt/wt) Gln-enriched enteral diet or an isonitrogenous isocaloric control diet for 14 days. Blood flow measurements were performed at day 16 using 46Sc-labeled microspheres. In the Gln-enriched group, higher organ blood flows were measured in the stomach (51%), the pancreas (35%), small intestine (32%), and colon (55%), compared with controls. No differences were found in systemic hemodynamic parameters between the control and Gln-supplemented groups. A possible role for nitric oxide in this splanchnic vasodilation was investigated. Daily urinary nitrate excretion was measured during the study but showed no significant differences between the control and Gln-fed animals. No differences were found in plasma levels of the vasodilating hormone glucagon between the groups. These results show that a Gln-enriched enteral diet increased splanchnic blood flow, which was not mediated by pancreatic glucagon or increased nitric oxide production as determined by urinary nitrate excretion.
Caco-2 cells express type I interleukin-1 receptors: ligand binding enhances proliferation
Authors: Varilek G. W. , Neil G. A. , Bishop W. P.
Published: December 2017

We examined the effect of interleukin-1 (IL-1) on the rate of proliferation of the human colon carcinoma Caco-2 and characterized the human intestinal epithelial cell IL-1 receptor (IL-1R). IL-1 dose dependently increased tritiated thymidine uptake in confluent Caco-2 monolayers fed complete growth medium. An anti-IL-1 beta completely blocked the increase in tritiated thymidine uptake, whereas an IL-1 receptor antagonist human recombinant blocked it partially. In long-term culture, IL-1 increased DNA content over control, an effect similar to that of epidermal growth factor (EGF). Unlike EGF, IL-1 did not enhance tritiated thymidine uptake in Caco-2 monolayers grown in serum-free medium, implying that IL-1 needs a cofactor(s) to elicit its proliferative effect. Cross-linking 125I-IL-1 beta to Caco-2 membranes revealed a binding protein of approximately 80 kDa with binding saturated at approximately 2.5 x 10(9) M-1 consistent with that for the type I IL-1R. cDNA transcribed from Caco-2 mRNA and amplified by polymerase chain reaction, using complementary oligonucleotides, resulted in a reaction product matching the sequence of the type I IL-1R. Our results demonstrate that IL-1 enhances proliferation of Caco-2 cells. This effect requires the presence of an unidentified cofactor(s). Also, Caco-2 cells express the type I IL-1R.
Actions of cholecystokinin and norepinephrine on vagal inputs to ganglion cells in guinea pig gallbladder
Authors: Mawe G. M. , Gokin A. P. , Wells D. G.
Published: December 2017

Previous studies have demonstrated that all guinea pig gallbladder neurons receive nicotinic synaptic input and that cholecystokinin (CCK) and norepinephrine have presynaptic facilitory and inhibitory effects, respectively, on these fast synaptic events. The current study was undertaken to determine the sources of the cholinergic terminals that provide nicotinic input to gallbladder neurons. To stimulate potential extrinsic inputs to gallbladder neurons, a stimulating electrode was placed on the nerve bundles that pass along the cystic duct. Stimulation of these nerves elicited fast excitatory postsynaptic potentials (EPSPs) in gallbladder neurons that were sensitive to hexamethonium, facilitated by CCK, and inhibited by norepinephrine. After vagotomy, most neurons (14 of 18) did not exhibit any nicotinic input. However, some neurons (3 of 18) did exhibit fast EPSPs in response to fiber tract stimulation, but not cystic nerve stimulation, indicating that interganglionic communication does exist amongst gallbladder neurons. These results demonstrate that the vagus nerves provide the major nicotinic input to gallbladder neurons. Furthermore, these data suggest that vagal terminals within gallbladder are a site of neurohormonal modulation of gallbladder ganglionic output by CCK, norepinephrine, and possibly other compounds.
Duodenal pH governs interdigestive motility in humans
Authors: Woodtli W. , Owyang C.
Published: December 2017

In this study, we examined the potential influence of duodenal pH in regulating the occurrence of the interdigestive migrating myoelectric complex (IMMC). Fasting gastroduodenal motility, duodenal pH, and plasma motilin were studied in 15 healthy subjects. During phase I, duodenal pH remained stable at 7 +/- 0.2. Phase II was accompanied by a lowering of duodenal pH, which fluctuated between 2.0 and 7.5. During late phase II, the duodenal pH increased to 6.9 +/- 0.3 and remained in the alkaline range during phase III. In six of 46 episodes of the IMMC, the occurrence of gastric phase III was delayed. This was associated with a persistently low duodenal pH (< 4) during late phase II. Despite a normal cyclic increase of plasma motilin, no gastric phase III activity was observed until the duodenal pH exceeded 7.0. Further studies showed that lowering of duodenal pH by intraduodenal perfusion of HCl prevented the occurrence of gastric phase III. We concluded that regularity of IMMC is governed by duodenal pH. An alkaline pH is essential for the initiation of gastric phase III; lowering of duodenal pH prevents its occurrence despite normal cyclic increase of plasma motilin.
Gastrin and carbachol require cAMP to elicit aminopyrine accumulation in isolated pig and rat parietal cells
Authors: Li Z. Q. , Cabero J. L. , Mardh S.
Published: December 2017

The role of endogenous adenosine 3',5'-cyclic monophosphate (cAMP) in the mechanisms of action of gastrin and carbachol on aminopyrine accumulation in isolated pig and rat parietal cells was investigated. In pig cells, pentagastrin (100 nM) alone stimulated aminopyrine accumulation, an action significantly reduced by the protein kinase A inhibitor Rp-adenosine 3',5'-cyclic monophosphothioate (Rp-cAMP[S]; 100 microM). In rat cells, gastrin-17 (100 nM) was incapable of stimulating aminopyrine accumulation, but it potentiated the action of histamine (100 microM). Carbachol (10 microM) stimulated aminopyrine accumulation and potentiated the action of histamine, and its action was potentiated in a dose-dependent manner by Sp-adenosine 3',5'-cyclic monophosphothioate (Sp-cAMP[S]; a cAMP analogue) in both species. The effect of carbachol was dose dependently reduced by Rp-cAMP[S]. The basal cAMP in pig parietal cells was 3.5-fold higher than that in rat parietal cells. Histamine (100 microM) and 3-isobutyl-1-methylxanthine (IBMX; 100 microM) only slightly elevated the cAMP content (1.2- to 2.9-fold the basal level) in both pig and rat parietal cells. Their combination, however, increased the cAMP level by 8- to 38-fold, but it did not increase aminopyrine accumulation above that elicited by histamine alone. Gastrin did not alter the cAMP levels in parietal cells of either of the two species. Both gastrin and carbachol increased cytosolic free Ca2+ in enriched pig and rat parietal cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Inhibition of gastric mechanoreceptor discharge by cholecystokinin in the rat
Authors: Grundy D. , Bagaev V. , Hillsley K.
Published: December 2017

The aim of the present study was to investigate electrophysiologically the effect of systemic cholecystokinin (CCK) on the discharge of vagal gastric mechanoreceptors. Twenty-two single vagal afferent fibers were selected for the investigation of responses to intravenous CCK octapeptide (CCK-8) on the basis of a positive response to gastric distension. Resting discharge in these afferent fibers was 1.3 +/- 0.3 impulses.s-1 and increased to 9.2 +/- 0.9 impulses.s-1 during distension (P < 0.0001), CCK (20-100 pmol iv) caused a gastric relaxation of 2.1 +/- 0.2 cmH2O and inhibition of phasic motility. The discharge of 20/22 of vagal tension receptors closely followed the magnitude and time course of the fall in pressure. Mean discharge before and after CCK (50 pmol) was 7 +/- 0.9 and 3.9 +/- 0.8 impulses.s-1, respectively (P < 0.001, n = 22). Both the pressure response and the concomitant changes in afferent discharge were abolished by L-364,718 (1.2 mg/kg iv). Only two afferent units failed to show a decrease in firing following CCK (50 pmol), and at 500 pmol the discharge of these units was augmented. In conclusion, CCK (50 pmol) has predominantly indirect effects on gastric mechanoreceptors, which decrease their firing in association with gastric relaxation.
Enteropathogenic Escherichia coli adherence to intestinal epithelial monolayers diminishes barrier function
Authors: Spitz J. , Yuhan R. , Koutsouris A. , Blatt C. , Alverdy J. , Hecht G.
Published: December 2017

The mechanism by which enteropathogenic Escherichia coli (EPEC) causes diarrhea remains elusive. Several alterations within the host cell have been demonstrated to occur following EPEC attachment including increases in intracellular Ca2+ concentration and rearrangement and phosphorylation of several cytoskeletal proteins. The consequences of these intracellular perturbations on host cell function, however, have not been determined. The aim of this study was to examine the effect of EPEC adherence on intestinal epithelial barrier function. T84 cell monolayers were infected with either wild-type EPEC or a nonadherent isogenic derivative. Transepithelial electrical resistance, a measure of barrier function, decreased 33.5 +/- 6.4% after a 6-h incubation with the wild-type strain. Electron microscopy revealed ultrastructurally normal cells, and lactate dehydrogenase release assays failed to demonstrate cytotoxicity. Dual 22Na+ and [3H]mannitol flux studies localized the permeability defect to tight junctions. In addition, cumulative flux of the paracellular marker mannitol was four- to fivefold greater across monolayers infected with wild-type EPEC. Sequestration of intracellular calcium stores by dantrolene completely abrogated the resistance drop associated with EPEC attachment. These data demonstrate that adherence of EPEC to intestinal epithelial cell monolayers disrupts tight junction barrier function via a calcium-requiring event.
Polyamines alter intestinal glucose transport
Authors: Johnson L. R. , Brockway P. D. , Madsen K. , Hardin J. A. , Gall D. G.
Published: December 2017

Polyamines are required for the growth of all eukaryotic cells. Enterocytes respond to luminal nutrients with large increases in polyamine synthesis, even though they are mature, nonproliferating cells. The role of polyamines in these cells is unknown. The current experiments examined whether polyamines affected intestinal transport of glucose, since absorption is the primary activity of enterocytes and since polyamines are known to affect membrane function and stability. Glucose transport was examined in rabbit brush-border membrane vesicles (BBMV). BBMV from rabbits given 5% alpha-difluoromethylornithine (DFMO) in their drinking water 24 h before they were killed transported significantly less glucose than control vesicles [38% decrease in maximal transport rate (Jmax)]. Orogastric administration of spermine, spermidine, or putrescine to DFMO-treated animals 24 h before they were killed prevented the decrease. In rabbits receiving only orogastric spermine, glucose transport was significantly increased (64% increase in Jmax), whereas in vivo spermidine and putrescine decreased Jmax. This increase in Jmax caused by in vivo administration of spermine was not dependent on protein synthesis. Addition of polyamines whether in vivo or in vitro decreased Michaelis constant in vesicles from control and DFMO-treated animals. The change in glucose transport induced by DFMO or polyamines was not related to altered membrane lipid composition or fluidity.(ABSTRACT TRUNCATED AT 250 WORDS)