bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential expression of the neurotensin gene in the developing rat and human gastrointestinal tract
Authors: Evers B. M. , Rajaraman S. , Chung D. H. , Townsend C. M. , Wang X. , Graves K. , Thompson J. C.
Published: December 2017

Neurotensin (NT) is an important hormone regulating gut motility, secretion, and growth. The purpose of this study was to determine the developmental expression of the NT/neuromedin N gene (NT/N) in the gut and pancreas of rats and humans. We found that NT/N expression, initially low in the fetal rat jejunum and ileum, is increased by postnatal day 3. This increase is independent of contact with luminal nutrients as demonstrated by elevated NT/N expression in rat jejunoileal grafts implanted in nude mice. NT/N expression reaches maximal levels in the small bowel by postnatal day 14. After postnatal day 28, NT/N mRNA levels remain constant in the ileum but decrease in the jejunum. Transient NT/N expression is found in the colon of fetal and postnatal rats. Similar to the rat, NT/N expression is low in the human fetal ileum but increases in the adult. In the human colon, NT/N is transiently expressed in the fetus at midgestation but disappears by birth and, similarly, is not apparent in the adult. We conclude the following. 1) The NT/N gene demonstrates a complex pattern of tissue-specific expression; the jejunum and ileum show a similar pattern of expression until the end of the fourth postnatal week, when NT/N levels decrease in the jejunum to assume the distinctive adult topographical distribution with NT/N increasing along the jejunoileal axis. 2) NT/N is transiently expressed in the colon of rats and humans during a developmental stage characterized by morphological and functional similarities to the small bowel; therefore, NT/N may provide a useful endocrine marker to further define the complex differentiation pathway leading to small bowel and colonic phenotypes.
Role of the enteric nervous system in the control of migrating spike complexes in the feline small intestine
Authors: De Vos W. C.
Published: December 2017

The effects of agonists and antagonists of nicotinic, muscarinic (M1 and M2), and adrenergic receptors on migrating spike complexes (MSC) in ileum of fasting cats are reported. Hexamethonium decreased MSC frequency and blocked propagation. Atropine at low concentrations increased MSC frequency and increased velocity of propagation; atropine at high concentration blocked propagation. Pirenzepine (Pz; M1 antagonist) increased MSC frequency and propagation velocity. McNeil A-343 (M1 agonist), by a Pz-sensitive phentolamine-insensitive mechanism, and 4-diethylamine-methylpiperidine (4-DAMP; M2 antagonist) blocked propagation of an ongoing MSC but had no significant effect on frequency or velocity. Bethanechol (M2-receptor agonist) increased phasic spiking by a 4-DAMP-sensitive mechanism and blocked MSC propagation by a Pz-sensitive mechanism. Phenylephrine (alpha 1-adrenoceptor agonist) or oxymetazoline (alpha 2-adrenoceptor agonist) blocked MSC propagation but had no effect on MSC frequency or velocity. Phentolamine (nonselective alpha 1-adrenoceptor antagonist), prazosin (alpha 1-adrenoceptor antagonist), or yohimbine (alpha 2-adrenoceptor antagonist) alone had no effect on MSC activity. The conclusion is that the enteric nervous system controls and regulates the MSC by the following proposed mechanisms. 1) M1-muscarinic receptors, located either on postganglionic inhibitory neurons or presynaptically at a nicotinic synapse and/or neuromuscular junction, are involved in the tonic inhibitory control of MSC initiation and propagation. 2) Nicotinic and M2 muscarinic receptors, located on excitatory postganglionic motoneurons and smooth muscle cells, respectively, are important in the initiation and/or propagation of MSC. 3) alpha 1-Adrenoceptors on the smooth muscle cells and alpha 2-adrenoceptors located presynaptically at the nicotinic ganglionic synapses are not tonically active but inhibit MSC activity (4). Smooth muscle beta-adrenoceptors do not play a significant role in neural control of MSC activity.
Proximal duodenal enterocyte transport: evidence for Na(+)-H+ and Cl(-)-HCO3- exchange and NaHCO3 cotransport
Authors: Isenberg J. I. , Ljungstrom M. , Safsten B. , Flemstrom G.
Published: December 2017

The duodenum, in contrast to the jejunum, actively secretes HCO3- at a high rate, a process that protects the mucosa from acid/peptic injury. Our purpose was to define the mechanisms involved in HCO3- transport by studying the acid-base transport processes in isolated duodenal enterocytes. Individual rat duodenocytes, isolated by a combination of Ca2+ chelation and collagenase, attached to a collagen matrix were loaded with the pH-sensitive fluoroprobe 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM), and intracellular pH was monitored by microfluorospectrophotometry. To identify Na(+)-H+ transport, cells in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid 1) were pulsed with NH4Cl (40 mM) in the absence and presence of amiloride and 2) were removed of Na+. To examine Cl(-)-HCO3- exchange, Cl- was removed from Ringer-HCO3- superfusate in the presence and absence of dihydro-4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (H2DIDS). The NaHCO3 cotransporter was studied by addition and subtraction of Na+ to amiloride-treated and Cl(-)-depleted enterocytes perfused with Na(+)- and Cl(-)-free Ringer-HCO3- buffer with and without H2DIDS. Mammalian duodenocytes contain at least three acid-base transporters: an amiloride-sensitive Na(+)-H+ exchanger that extrudes acid, a DIDS-sensitive Cl(-)-HCO3- exchanger that extrudes base, and a NaHCO3 cotransporter, also DIDS sensitive, that functions as a base loader. These acid-base transporters likely play a key role in duodenal mucosal HCO3- secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
Modulation of uptake of heme by rat small intestinal mucosa in iron deficiency
Authors: Roberts S. K. , Henderson R. W. , Young G. P.
Published: December 2017

Although body iron status modulates whole body retention of heme derived iron, it is not known with certainty whether modulation occurs by regulation of mucosal uptake of heme. In vivo uptake from perfused intestine of heme labeled with 14C in the porphyrin ring was studied in groups of rats of differing iron status ranging from fully replete to markedly iron deficient. Heme extraction from infusate and mucosal heme uptake were significantly different between test groups (P < 0.005 and 0.001, respectively). Marked iron deficiency induced a 4.8-fold rise in heme extraction relative to iron-replete animals; rats with latent iron deficiency showed a smaller but still significant rise. Heme extraction correlated negatively with indicators of iron status: hemoglobin (r = -0.76, P < 0.001) and serum iron (r = -0.56, P < 0.05). The specific binding of [14C]heme to purified brush borders from iron-replete and iron-deficient rats was 1.4-fold higher in deficient rats when expressed per milligram of protein (P = 0.046) and 3.3-fold higher when expressed relative to alkaline phosphatase activity (P = 0.014). Thus mucosal uptake of heme in iron deficiency is increased because of an increase in its binding to the brush border.
Inhibitory effect of CO on internal anal sphincter: heme oxygenase inhibitor inhibits NANC relaxation
Authors: Rattan S. , Chakder S.
Published: December 2017

We examined the effect and role of CO in opossum internal anal sphincter (IAS) relaxation in response to nonadrenergic noncholinergic (NANC) nerve stimulation. Effects of NANC nerve stimulation on the IAS tension and second messengers (cAMP and cGMP) were examined before and after the selective heme oxygenase (HO) inhibitor zinc protoporphyrin IX (Zn PP-IX). The HO activity of the IAS smooth muscle was determined before and after NANC nerve stimulation. CO caused a concentration-dependent and tetrodotoxin-resistant fall in the resting tension of the IAS. The direct action of CO was confirmed by its relaxant action on the isolated smooth muscle cells. Furthermore, CO caused an increase in the tissue cGMP levels comparable to that observed with nerve stimulation. Zn PP-IX caused suppression of IAS relaxation caused by NANC nerve stimulation and vasoactive intestinal polypeptide (VIP) but not by peptide histidine-isoleucine and suppression of the increase in cGMP in response to NANC nerve stimulation. Zn PP-IX had no significant effect on the IAS responses to CO, nitric oxide (NO), and the beta-adrenoceptor agonist isoproterenol. The IAS responses to CO were not modified by the NO synthase inhibitor NG-nitro-L-arginine. Significant HO activity was detected in the IAS, which increased further in response to NANC nerve stimulation and VIP. The direct relaxant actions of CO and the suppression of NANC-mediated relaxation of the IAS by the HO inhibitor suggest the involvement of CO in the neurally mediated IAS relaxation.
A novel in vitro technique to study extrinsic neural control of rabbit colonic muscle and epithelial function
Authors: Goldhill J. M. , Percy W. H.
Published: December 2017

A novel in vitro technique capable of simultaneously measuring distal colonic epithelial potential difference and muscle contraction is described. Under basal conditions, oscillations in both muscle tone and potential difference were observed. Pelvic nerve stimulation was shown to evoke strong "duration" contractile responses in both the longitudinal and circular muscle layers. Additionally, tonic changes in potential difference extending beyond the train of stimuli were observed, suggesting for the first time that colonic ion transport may be influenced by the pelvic nerves. However, it was unclear whether these were direct effects or indirect actions resulting from muscle contractions causing mechanical stimulation of nerves of the submucosal plexus. Lumbar colonic nerve stimulation inhibited spontaneous contractile activity and reduced basal tone in both muscle layers. However, there was no consistent effect of sympathetic nerve stimulation on transepithelial potential difference. Each of the muscle and epithelial effects of sympathetic nerve stimulation was mimicked by exogenous norepinephrine. Based on these data, it is concluded that colonic function is strongly influenced by the extrinsic innervation. Furthermore, relatively long-term modulation of epithelial function can be achieved by short bursts of pelvic nerve activity.
Bidirectional membrane transport of intact glutathione in Hep G2 cells
Authors: Sze G. , Kaplowitz N. , Ookhtens M. , Lu S. C.
Published: December 2017

Rat hepatocytes exhibit bidirectional carrier-mediated transport of reduced glutathione (GSH) across the plasma membrane. Transport of GSH has not been well characterized in human-derived cells. We examined Hep G2 cells as a possible human liver model for GSH homeostasis. Hep G2 cell GSH averaged 25.9 +/- 1.4 nmol/10(6) cells. When Hep G2 cells were incubated in buffer, no GSH appeared in the medium over 2 h. However, after pretreatment with acivicin to inhibit gamma-glutamyl transpeptidase activity, GSH efflux was unmasked and measured 30 +/- 4 pmol x 10(6) cells-1 x min-1, which is comparable to rat hepatocytes. GSH efflux was inhibited by sulfobromophthalein GSH adduct (BSP-GSH) and cystathionine, agents that inhibit sinusoidal efflux in the rat, and was stimulated by adenosine 3',5'-cyclic monophosphate-dependent agents. GSH uptake was measured after cells were pretreated with acivicin and buthionine sulfoximine to prevent breakdown of GSH and resynthesis of GSH from precursors, respectively. In the presence of 4 microCi/ml of [35S]GSH and 10 mM unlabeled GSH, GSH uptake was linear up to 45 min and did not require Na+ or Cl-. GSH uptake exhibited saturability with a maximal velocity of 4.15 +/- 0.23 nmol.mg-1 x 30 min-1, a Michaelis constant of 2.36 +/- 0.26 mM, and two interactive transport sites. BSP-GSH cis-inhibited GSH uptake in a dose-dependent manner with an inhibitory constant of 0.46 +/- 0.05 mM. Inhibition by BSP-GSH (1 mM) of GSH uptake was through a single inhibitor site and was overcome at > 10 mM GSH, which is consistent with competitive inhibition. Similar to the rat, 10 mM extracellular GSH trans-stimulated GSH efflux. These findings may be important in gaining better insights into GSH homeostasis in human liver cells.
Pancreatic polypeptide in dorsal vagal complex stimulates gastric acid secretion and motility in rats
Authors: McTigue D. M. , Edwards N. K. , Rogers R. C.
Published: December 2017

High concentrations of receptors for pancreatic polypeptide (PP), a pancreatic hormone, were recently discovered in the dorsomedial region of the dorsal vagal complex (DVC). We hypothesized that gastric acid secretion and motility, digestive functions strongly influenced by vagovagal reflexes organized within the DVC, would be affected by PP applied directly to this vagal sensorimotor integration area. After urethan-anesthetized rats were prepared for antral motility recording or titrometric analysis of gastric acid output, phosphate-buffered saline or various doses of PP in phosphate-buffered saline were micropressure injected into the medial DVC. Injections of PP into the DVC produced significant, long-lasting, and dose-dependent increases in gastric acid secretion and antral motility. These gastric responses were blocked by bilateral cervical vagotomy and by atropine, suggesting that intramedullary PP stimulates vagal cholinergic pathways, resulting in enhanced gastric functions. Because PP is not synthesized within the central nervous system, these results point to a new mechanism whereby the digestive tract may modulate its own autonomic control: direct humoral action on vagovagal reflex circuits within the brain stem.
Galanin receptor in plasma membrane of canine small intestinal circular muscle
Authors: Chen C. K. , McDonald T. J. , Daniel E. E.
Published: December 2017

High-affinity binding sites for galanin were identified and characterized in plasma membrane of circular muscle from canine small intestine using 125I-radioiodinated synthetic porcine galanin. Scatchard analysis indicated a high-affinity binding site on plasma membrane with a dissociation constant (Kd) of 0.58 nM and a binding capacity of 389 fmol/mg. Unlabeled galanin or NH2-terminal galanin fragments competitively inhibited the binding of 125I-galanin in a concentration-dependent manner, whereas the COOH-terminal fragment was inactive. Computer analysis of competitive binding data suggested a two-site model with a high-affinity (inhibitor constant, Ki = 0.01 nM) and a low-affinity (Ki = 2.8 nM) binding site. Guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) enhanced the dissociation of bound 125I-galanin. Cholera toxin (CTX) and GTP gamma S abolished the activity of the high-affinity binding site, leaving the low-affinity binding site. We conclude that galanin may act as an neurotransmitter to inhibit canine small intestinal smooth muscle contraction by interaction with a CTX-sensitive G protein-coupled specific receptor on muscle membrane. This receptor showed different G protein coupling from a synaptosomal receptor previously described in the same tissue preparations
Endothelin-1 stimulates bile acid secretion and vesicular transport in the isolated perfused rat liver
Authors: Tanaka A. , Katagiri K. , Hoshino M. , Hayakawa T. , Tsukada K. , Takeuchi T.
Published: December 2017

The effects of endothelin (ET) on portal pressure and bile secretion were examined using isolated perfused rat liver and rat hepatocyte preparations. ET-1 raised portal pressure dose dependently; administration at a high dose (10(-9) mol) induced a > 200% increase along with reduced bile flow and decreased secretion of bile acid and phospholipids. However, a low dose (10(-10) mol) of ET-1 brought about a < 100% portal pressure rise, enhanced both bile flow and excretion of bile acid and phospholipids, and significantly increased transfer of preadministered horseradish peroxidase (HRP) into bile. In addition, values for Ca2+ concentrations, examined by indo 1 fluorescence, were elevated in isolated hepatocytes after administration of ET-1. Papaverine suppressed the low-dose ET-1 stimulation effects on both portal pressure and bile secretion. Moreover, it also reduced the HRP excretion and suppressed intracellular Ca2+ release. This study demonstrated that ET-1 stimulates vesicular transport, probably via promotion of intracellular Ca2+ release, and, as a result, increases bile acid-dependent bile flow.
Incomplete processing of progastrin expressed by human colon cancer cells: role of noncarboxyamidated gastrins
Authors: Singh P. , Xu Z. , Dai B. , Rajaraman S. , Rubin N. , Dhruva B.
Published: December 2017

Gastrin is mitogenic for several colon cancers. To assess a possible autocrine role of gastrin in colon cancers, we examined human colon cancer cell lines for expression of gastrin mRNA and various forms of gastrin. Gastrin mRNA was not detected in the majority of colon cancer cell lines by Northern hybridization but was detected in all human colon cancer lines by the sensitive method of reverse transcriptase-polymerase chain reaction (PCR). Gastrin mRNA was quantitated by the competitive PCR method. The majority of cell lines expressed very low levels of gastrin mRNA (< 1-5 copies/cell); only one cell line expressed > 20 copies/cell. The mature carboxyamidated form of gastrin was not detected in any of the cell lines by radioimmunoassay or immunocytochemistry. Results suggested that either gastrin mRNA expressed by colon cancer cells was altered (mutated) or posttranslational processing of progastrin was incomplete. Gastrin cDNA from all the colon cancer cell lines had an identical sequence to the published sequence of human gastrin cDNA. Specific antibodies against precursor forms of gastrin were used, and significant concentrations of nonamidated (glycine-extended) and prepro forms of gastrin were measured in tumor extracts of representative colon cancer cell lines. The presence of precursor forms of gastrin suggested a lack of one or more of the processing enzymes and/or cofactors. Significant concentrations of the processing enzyme (peptidylglycine alpha-amidating monooxygenase) were detected in colon cancer cells by immunocytochemistry. Therefore, lack of other cofactors or enzymes may be contributing to incomplete processing of precursor forms of gastrin, which merits further investigation. Since low levels of gastrin mRNA were expressed by the majority of human colon cancer cell lines and progastrin was incompletely processed, it seems unlikely that gastrin can function as a viable autocrine growth factor for colon cancer cells. High concentrations of glycine-extended gastrin-17 (GG) (> 10(-6) M) were mitogenic for a gastrin-responsive human colon cancer (DLD-1) cell line in vitro. It remains to be seen if GG or other precursor forms of gastrin are similarly mitogenic in vivo, which may then lend credibility to a possible autocrine role of gastrinlike peptides in colon cancers.
Rapid upregulation of snake intestine in response to feeding: a new model of intestinal adaptation
Authors: Secor S. M. , Stein E. D. , Diamond J.
Published: December 2017

Mammalian guts exhibit numerous adaptive responses to feeding. However, response magnitudes are often inconveniently modest for experimental analysis, because mammals feed often and their intestines are rarely empty. We anticipated larger responses in sit-and-wait foraging snakes, because they consume huge meals at long intervals. Hence, we studied metabolic rates, brush-border nutrient transport, and intestinal morphometrics in the rattlesnake, Crotalus cerastes, as a function of time since feeding. O2 consumption by the whole snake, a reflection of the cost of digestion and of rebuilding the starved gut, peaked after 2 days at eight times fasting values. Activities of brush-border glucose, leucine, and proline transporters peaked after 1-3 days at 5-22 times fasting values. Ratios of amino acid to glucose uptake rates peaked at 104, reflecting snakes' extreme adaptation to carnivory (a high-protein low-carbohydrate diet). Intestinal mass increased more than twofold within 1 day, primarily because of mucosal growth. After defecation, the intestine atrophied, brush-border transporters were downregulated, and O2 consumption returned to basal. These rapid and large responses reduce costs of gut maintenance during long bouts of quiescence between meals. Hence sit-and-wait foraging snakes may furnish advantageous model species for studying gut regulation and adaptation.
Inhibiting gastric H(+)-K(+)-ATPase activity by omeprazole promotes degeneration and production of parietal cells
Authors: Karam S. M. , Forte J. G.
Published: December 2017

Parietal cell morphology was studied after chronic inhibition of gastric H(+)-K(+)-adenosinetriphosphatase (ATPase) by omeprazole. Gastric mucosa from rabbits treated with omeprazole every 12 h (1 mg/kg sc) for 5 days were compared with sham-injected animals using immunohistochemistry and electron microscopy. Three immunocytochemical markers, including antibodies against the alpha- and beta-subunits of the H(+)-K(+)-ATPase, showed that some parietal cells in omeprazole-treated rabbits displayed light areas and granularity in their cytoplasm. These abnormalities were also apparent in semithin sections. Electron microscopy was used to categorize and quantitate the specific structural abnormalities in parietal cells. Cells were classified as normal, altered, or degenerated. For control tissues, altered and degenerated parietal cells were few; they collectively represented 6% of total parietal cells and were located mainly deep in the gland base. For omeprazole-treated tissues, altered and degenerated parietal cells occurred throughout the gland and averaged 62% of total parietal cells. In addition, macrophages invaded the mucosa presumably to eliminate degenerated cells. Although there was an increase in parietal cell degeneration, enhanced parietal cell generation was suggested by increases in mitosis among proliferative cells and, more specifically, in the number of preparietal cells. After 3 days of recovery from the omeprazole regimen, parietal cells and the gastric mucosa appeared to recover the normal morphology. In conclusion, blocking H(+)-K(+)-ATPase by omeprazole enhances degeneration and macrophage-mediated elimination of parietal cells and also causes an increase in preparietal cell production. Thus omeprazole temporarily changes the dynamic features of parietal cells in the rabbit to make them die early and grow fast.
Stimulation of rat parietal cell function by histamine and GLP-1-(7-36) amide is mediated by Gs alpha
Authors: Schmidtler J. , Dehne K. , Offermanns S. , Rosenthal W. , Classen M. , Schepp W.
Published: December 2017

It was the aim of the present study to determine in rat parietal cells whether Gs alpha, the stimulatory subunit of adenylate cyclase, mediates adenosine 3',5'-cyclic monophosphate (cAMP)-dependent parietal cell function in response to histamine and glucagon-like peptide 1 (GLP-1)-(7-36) amide. Cytoplasmic membrane from enriched (83 +/- 5%) rat parietal cells were incubated for 30 min with 30 microCi/ml [32P]NAD+ and 40 micrograms/ml preactivated cholera toxin (CT), a pharmacological tool for activation of Gs alpha. Subsequent sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography revealed [32P]ADP ribosylation of Gs alpha represented by three proteins with molecular masses ranging from 42 to 45 kDa. In intact parietal cells, CT (10(-12)-10(-8) M) caused marked stimulation of [14C]aminopyrine accumulation and cAMP production confirming the functional importance of Gs alpha in regulation of H+ production. Identical membrane preparations were preincubated (2 h, 4 degrees C) in parallel with and without RM/1, a rabbit polyclonal anti-Gs alpha-antibody. Subsequently, adenylate cyclase was stimulated by histamine, GLP-1-(7-36) amide, CT, or forskolin. At a 1:10 dilution, the antiserum completely abolished adenylate cyclase activity in response to maximal concentrations of histamine, GLP-1-(7-36) amide, and CT while reducing forskolin stimulation by only 22.0 +/- 4.9%. At 1:50, RM/1 reduced responses to histamine, GLP-1-(7-36) amide and CT by 20-30% but failed to inhibit forskolin-stimulated enzyme activity. At 1:100, the antiserum was ineffective versus all stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)
Mast cell activation augments gastric mucosal injury through a leukotriene-dependent mechanism
Authors: Rioux K. P. , Wallace J. L.
Published: December 2017

Several lines of evidence suggest a role for mast cells as modulators of gastric mucosal integrity, but the effect of antigenic mast cell activation on mucosal resistance to injury has not previously been examined. In this study, rats were sensitized to the nematode Nippostrongylus brasiliensis and were studied 35-42 days later. With use of an ex vivo gastric chamber preparation, the stomach was exposed for 10 min to 20% ethanol. In some rats, antigen was administered intra-arterially 10 min before application of ethanol. Sensitized rats exhibited similar levels of ethanol-induced gastric injury as control rats, despite having significantly greater numbers of mucosal mast cells. However, antigen administration, which did not in itself produce mucosal injury, significantly augmented (approximately 3-fold) the extent of injury in sensitized but not control rats. Prior treatment with dexamethasone depleted mucosal mast cells in control and sensitized rats. Moreover, this treatment abolished the increase in mucosal injury observed in sensitized rats treated with antigen and topical ethanol. Pretreatment with a leukotriene D4-receptor antagonist, but not a platelet-activating factor-receptor antagonist or a cyclooxygenase inhibitor, abolished the increased susceptibility of sensitized rats to gastric damage induced by antigen and topical ethanol. These results suggest that mucosal mast cell number per se does not influence mucosal susceptibility to injury; however, activation of mast cells markedly increases the susceptibility to injury through a peptidoleukotriene-dependent mechanism.
Control of canine gastric emptying of fat by lipolytic products
Authors: Meyer J. H. , Elashoff J. D. , Domeck M. , Levy A. , Jehn D. , Hlinka M. , Lake R. , Graham L. S. , Gu Y. G.
Published: December 2017

Dietary fat is ingested in three forms: 1) in solid food, 2) as aqueous emulsions, and 3) as unemulsified, liquid oil. On the basis of a scant previous literature, we postulated that liquid fat (emulsions or oils) would empty from the stomach at speeds that varied with the amounts ingested but that this dynamic would be modulated by feedback inhibition from lipolytic products. To test these ideas, we used a gamma camera to track gastric emptying of 123I-labeled fat in dogs with chronic pancreatic fistulas by which lipase was excluded from or replenished in the duodenum in varied amounts after dogs were fed 15-, 30-, and 60-g loads of liquid fat given with solid foods or as emulsions. We also tracked concurrent gastric emptying of 113mIn, which marked the solid food phase or the water phase of emulsions. In some studies, we used a potent and specific inhibitor (orlistat) of pancreatic and gastric lipases to assess how lipolytic products modulated emptying of liquid fat. In the absence of pancreatic enzymes, both oils and emulsions emptied initially at high speeds that varied with fat loads, but emptying slowed 20 min after ingestion of emulsions and 60 min after ingestion of unemulsified oil. Studies with orlistat indicated that these changes in rates resulted from liberation of gastric lipolytic products. Emptying of oil emulsions was not altered by duodenal replenishment with pancreatic enzymes, but emptying of unemulsified oil was inhibited in a dose-related fashion, such that maximal inhibition was achieved when pancreatic enzymes were replenished at > or = 40% of normal amounts. Studies with orlistat confirmed that this dose-dependent slowing was due specifically to lipase. Emptying of solid food was much more sensitive to replenishment with enzymes, so that a 10% replenishment maximally inhibited solid emptying.
K+ transport in isolated guinea pig colonocytes: evidence for Na(+)-independent ouabain-sensitive K+ pump
Authors: Del Castillo J. R. , Sulbaran-Carrasco M. C. , Burguillos L.
Published: December 2017

K+ transport mechanisms in epithelial cells isolated from guinea pig distal colon have been studied using 86Rb as a tracer. A transport pathway has been identified that is proposed to be identical to the mechanism mediating transepithelial K+ absorption. Guinea pig colonocytes take up K+ through at least three separate mechanisms: 1) a Na(+)-dependent, ouabain-sensitive influx that is consistent with the Na(+)-K+ pump, 2) a Na(+)-dependent bumetanide-sensitive influx consistent with the Na(+)-K(+)-2Cl- cotransporter, and 3) a Na(+)-independent ouabain-sensitive influx, consistent with an apical colonic K+ pump. These transport mechanisms are sensitive to metabolic inhibition by rotenone and to vanadate, a blocker of type P adenosinetriphosphatase (ATPases). SCH-28080, an inhibitor of gastric K(+)-H(+)-ATPase, was without effect. Measurements of net K+ fluxes revealed that isolated colonocytes concentrated K+ by two processes: 1) a Na(+)-dependent ouabain-sensitive mechanism, which is compatible with the Na(+)-K+ pump and 2) a Na(+)-independent ouabain-sensitive mechanism consistent with the proposed absorptive K+ pump. These concentrative mechanisms were also inhibited by rotenone and vanadate, but not by SCH-28080. The Na(+)-independent ouabain-sensitive K+ pump was present in the distal colon, but absent in the proximal colon and the small intestine of guinea pig. It is proposed that this Na(+)-independent ouabain-sensitive K+ pump mediates K+ absorption and is related to the luminal K(+)-ATPase.
Prolonged administration of IGF peptides enhances growth of gastrointestinal tissues in normal rats
Authors: Steeb C. B. , Trahair J. F. , Tomas F. M. , Read L. C.
Published: December 2017

To investigate the effect of insulin-like growth factor (IGF) peptide infusion on the gastrointestinal tract, female rats (115 g, 6/group) were treated for 14 days with IGF-I or long R (LR3IGF-I; 0, 44, 111, or 278 micrograms/day) delivered by osmotic minipumps. Both peptides induced a dose-dependent increase in gastrointestinal tissue weight. Total gut weight, small intestinal weight, and small intestinal length increased by 43, 47, and 13%, respectively, after treatment with 278 micrograms/day of LR3IGF-I. Crypt depth and villus height increased after peptide treatment with an associated increased crypt cell population (+33%), cells per villus column (+34%), and villus cell density (+20%). Proportional increments in proliferating cell nuclear antigen labeling and an unaltered crypt growth fraction indicated that the balance between the proliferative and maturation compartment of the crypt was maintained. Fecal nitrogen excretion was significantly reduced in rats treated with LR3IGF-I, suggesting an increased absorptive capacity of the duodenum. The enhanced potency of LR3IGF-I supports previous findings that the gut is especially responsive to analogues with reduced binding affinity to IGF-binding proteins.
CGRP as a transmitter in the sensory pathway mediating peristaltic reflex
Authors: Grider J. R.
Published: December 2017

The presence of calcitonin gene-related peptide (CGRP) in enteric and dorsal root ganglion neurons suggests that it might be involved as a transmitter in the sensory pathway that mediates the peristaltic reflex. This notion was examined in a three-compartment flat-sheet colonic preparation in which stimuli (muscle stretch or mucosal stimulation) were applied to the central compartment, while ascending contraction and descending relaxation were measured in the peripheral compartments. Both muscle stretch and mucosal stroking applied to the central compartment elicited CGRP release into the central, but not peripheral, compartments. The magnitude of release was proportional to the intensity of stimulation. Addition of the CGRP antagonists, human CGRP-(8-37) and [Tyr0]CGRP-(28-37), to the central compartment inhibited descending and ascending responses elicited by muscle stretch or mucosal stimulation. Addition of the sensory neurotoxin, capsaicin (0.01 to 1 microM), to the central compartment caused concentration-dependent release of CGRP; the resultant depletion of sensory transmitter resulted in a concentration-dependent decrease in ascending and descending responses elicited by muscle stretch and mucosal stimulation. Extrinsic denervation decreased basal CGRP release and abolished CGRP release and peristaltic responses normally elicited by muscle stretch but not CGRP release or peristaltic responses elicited by mucosal stimulation. The results indicate that extrinsic sensory pathways, which mediate the peristaltic response to muscle stretch, and intrinsic sensory pathways, which mediate the peristaltic response to mucosal stimulation, utilize CGRP as a sensory transmitter.
Sympathetic input to ganglia of the guinea pig sphincter of Oddi
Authors: Wells D. G. , Mawe G. M.
Published: December 2017

Intracellular recording and immunohistochemical staining techniques were used to establish whether sphincter of Oddi (SO) ganglia are a target of sympathetic input to this region. Norepinephrine (0.01-10.0 microM) decreased the amplitude of the nicotinic fast excitatory postsynaptic potential (EPSP) evoked by stimulation of interganglionic fiber tracts, with a half-maximal inhibitory concentration (EC50) of 300 nM. Norepinephrine did not alter the responsiveness of the neurons to acetylcholine. The alpha 2-adrenoreceptor agonist UK-14304 mimicked the norepinephrine-induced effect with a EC50 of 2.5 nM, whereas alpha 1- and beta-adrenoreceptor agonists had no effect on the EPSP. The alpha 2-adrenoreceptor antagonist idazoxan (1.0 microM) inhibited the UK-14304 response, with a dissociation constant of 1.0 nM. Release of endogenous catecholamines, by the addition of tyramine (100 microM) to the bath, caused an idazoxan-sensitive decrease in the amplitude of the fast EPSP. In the minority of SO neurons that exhibited inhibitory postsynaptic potentials (IPSPs), norepinephrine caused a hyperpolarization of the membrane potential. The IPSP and the norepinephrine-induced hyperpolarization were inhibited by alpha 2-adrenoreceptor antagonists. Desipramine (1.0 microM), an uptake inhibitor, reversibly increased the amplitude of the IPSP. Immunoreactivities for tyrosine hydroxylase and dopamine beta-hydroxylase were coexistent in nerve fibers and nonexistent in cell bodies in the ganglionated plexus of the SO. The results of this study indicate that norepinephrine acts pre- and postsynaptically as an inhibitory neurotransmitter in SO ganglia.
Lectins are sensitive tools for defining the differentiation programs of mouse gut epithelial cell lineages
Authors: Falk P. , Roth K. A. , Gordon J. I.
Published: December 2017

We have used histochemical methods to survey the cellular patterns of binding of a panel of 45 lectins with well-defined carbohydrate specificities to sections prepared from various regions of the gastric-to-colonic axis of fetal, neonatal, and adult FVB/N mouse gut. The results suggest that lectins can be used as remarkably sensitive tools to describe the differentiation programs of gastric and intestinal epithelial cell lineages as a function of their position along the cephalocaudal axis of the gut and as a function of developmental stage. Studies of intestinal isografts and transgenic mice that express Simian virus-40 T antigen in enterocytes suggest that many of these cell lineage-specific and spatial patterns of glycoconjugate production can be established and maintained in the absence of exposure to luminal contents and in the presence of specific proliferative abnormalities. This lectin panel should be useful for operationally defining subpopulations of the principal gut epithelial cell lineages in normal strains of mice, for describing variations in gut epithelial cell differentiation programs in mutant and transgenic mice, and for recovering specific epithelial cell lineages or subpopulations.
Stimulation of esophageal mechanoreceptors is dependent on rate and duration of distension
Authors: Nguyen P. , Castell D. O.
Published: December 2017

We randomly compared rapid (170 ml/s) and slow (0.7 ml/s) rates of balloon distension in 19 volunteers (9 male, 10 female) using a latex balloon (length 3 cm) positioned 5 cm proximal to the lower esophageal sphincter. The balloon was inflated/deflated (1-s dwell; 5- to 10-s interval) with increasing 2-ml volumes during rapid inflation studies and progressively distended during slow inflation studies while subjects blindly indicated feeling, rated as follows: 0 (no sensation); 1 (aware of balloon, but no discomfort); 2 (pain or discomfort). In a second set of studies, the balloon was left inflated for 60 s at a volume 2 ml below that producing sensation 1, and time until a change in sensation was recorded. This was repeated at a volume 2 ml below that producing sensation 2. Mean volume to sensation 1 with slow inflation was 14.6 +/- 1.2 ml (SE) and with rapid inflation was 8.5 +/- 0.8 ml (P < 0.001). Mean volume to sensation 2 with slow inflation was 23.7 +/- 1.0 ml and with rapid inflation was 17.4 +/- 1.0 ml (P < 0.001). With sustained (60 s) inflation, most subjects noted increasing sensation, independent of initial inflation rate. We conclude that distal esophageal sensory perception (both awareness and discomfort) is dependent on rate of stimulation of mechanoreceptors. A sustained distension stimulus appears to recruit additional receptors and increase the level of sensory awareness.
Biliary excretion of plasma non-transferrin-bound iron in rats: pathogenetic importance in iron-overload disorders
Authors: Brissot P. , Zanninelli G. , Guyader D. , Zeind J. , Gollan J.
Published: December 2017

Plasma non-transferrin-bound iron (NTB-iron) is a potentially toxic form of iron, which is efficiently taken up by the normal, as well as the chronically iron-overloaded liver. In fact, NTB-iron may represent the major source of iron gaining access to hepatocytes in the iron-loaded state. We postulated that efficient biliary excretion of this form of iron could protect against iron-related hepatocellular injury. To characterize the biliary excretion of NTB-iron in intact normal and iron-loaded rats, the plasma disappearance and biliary excretion kinetics of plasma 55Fe-labeled NTB-iron were determined. In normal rats, prompt biliary excretion of plasma NTB-iron was evident, with peak radioactivity approximately 10 min after 55Fe injection (4.1% mean recovery at 3 h). In contrast, biliary iron excretion in iron-loaded rats was minimal (0.1%). In normal rats, a marked increase in biliary excretion of plasma NTB-iron was observed after intravenous desferrioxamine (mean recovery 20.9%) and the new oral iron chelator 1,2-dimethyl-3-hydroxypyrid-4-one administered intravenously (mean recovery 16.1%) or orally (11.4%). In iron-loaded rats, the cumulative recoveries of 55Fe in bile achieved by chelators were lower than in controls (7.6, 3.9, and 3.7%, respectively). Collectively, these findings demonstrate that 1) the normal liver rapidly excretes significant amounts of plasma NTB-iron in bile; 2) the iron-loaded liver exhibits a marked decrease in the capacity to excrete plasma NTB-iron into bile; and 3) chelating agents greatly enhance the biliary excretion of plasma NTB-iron, although the response in terms of cumulative recoveries is less pronounced in the iron-loaded state.(ABSTRACT TRUNCATED AT 250 WORDS)
ATP-MgCl2 administration normalizes macrophage cAMP and beta-adrenergic receptors after hemorrhage and resuscitation
Authors: Wang P. , Tait S. M. , Ba Z. F. , Chaudry I. H.
Published: December 2017

Although ATP-MgCl2 attenuates the release of inflammatory cytokines and restores the defective macrophage (M phi) antigen presentation function after hemorrhage and resuscitation, it is not known whether administration of this agent after hemorrhage affects M phi adenosine 3',5'-cyclic monophosphate (cAMP) levels and beta-adrenergic receptors. To determine this, rats underwent a midline laparotomy (i.e., induction of trauma) and were then bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of maximum bleedout volume was returned in the form of Ringer lactate (RL). Animals were resuscitated with four times the volume of shed blood with RL, during and after which ATP-MgCl2 (50 mumol/kg) or saline was administered over 95 min. At 1.5 h postresuscitation (i.e., 10 min after completion of ATP-MgCl2 infusion), peritoneal M phi and Kupffer cells were isolated, and cAMP levels were measured by radioimmunoassay. beta-Receptor binding characteristics were also determined in isolated Kupffer cells. The results indicate that cAMP levels increased significantly in both peritoneal M phi and Kupffer cells after hemorrhage and resuscitation. Maximum binding capacity (Bmax) of beta-receptors increased in Kupffer cells, suggesting that the elevated cAMP may be due to the increased beta-receptor Bmax under such conditions. ATP-MgCl2 treatment, however, markedly decreased beta-receptor Bmax in Kupffer cells and cAMP in both peritoneal M phi and Kupffer cells, and the values were similar to shams. Thus normalization of M phi cAMP levels and beta-receptor binding capacity by ATP-MgCl2 may contribute to the immunoenhancing effects of this agent observed after trauma-hemorrhage and fluid resuscitation.
Effect of age on induction of hepatic phosphoenolpyruvate carboxykinase by fasting
Authors: Van Remmen H. , Ward W. F.
Published: December 2017

This study examines the effect of age on the induction of the rate-limiting enzyme in gluconeogenesis, phosphoenolpyruvate carboxykinase (PEPCK), in response to fasting and refeeding in male Fischer 344 rats aged 3-18 mo. The rats were fasted for 30 h to increase the activity of PEPCK and subsequently were refed for 24 h to lower activity toward basal levels. PEPCK activity increased 2.2-fold in the 3-mo-old rats and 2.3-fold in the 18-mo-old rats during the 30-h fast. Therefore PEPCK induction during the 30-h fast was not altered with age. Similarly, refeeding resulted in a significant decrease in PEPCK activity at all ages. After the 24-h refeeding period, the rats were fasted a second time, and the time course of induction from the basal refed level was measured. In the young rats (6 mo), the activity of PEPCK increased rapidly from 18.12 +/- 1.61 to 42.66 +/- 5.94 U/g protein (P < 0.01) within 8 h of fasting. However, in the 18-mo-old rats, the initiation of the induction of PEPCK activity was delayed, and, after 12 h, PEPCK activity had increased from 17.34 +/- 1.34 to only 32.50 +/- 3.21 U/g protein (P < 0.01). Furthermore, the rate of induction appears to be decreased in the older animals. The activity after 24 h of fasting was equivalent in all four age groups (ranging from 44.72 +/- 5.38 at 3 mo to 40.18 +/- 5.42 U/g protein at 18 mo).(ABSTRACT TRUNCATED AT 250 WORDS)
Mechanism of cholinergic stimulation of glucose oxidation in isolated gastric glands
Authors: Hernandez I. , Chacin J.
Published: December 2017

The mechanisms of cholinergic activation of carbohydrate metabolism were investigated in isolated rabbit gastric glands. Carbachol stimulated the rate of glucose oxidation in a dose-dependent fashion with a half-maximal effect occurring at approximately 9 microM. Atropine and omeprazole, but not cimetidine, completely blocked the stimulation induced by carbachol. Direct activation of the H(+)-K(+)-adenosinetriphosphatase by NH+4 caused a significant stimulation of glucose oxidation that was totally abolished by oligomycin and by the mitochondrial uncouplers dinitrophenol and carbonyl cyanide p-trifluoromethoxyphenylhydrazone. These latter agents did not abolish the stimulating effect of carbachol on glucose oxidation. Ionomycin increased the rate of glucose oxidation in a dose-dependent manner, and this effect was not blocked by oligomycin. The metabolic effect of ionomycin was reduced but not abolished by omeprazole. 1,2-Bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester eliminated the carbachol-induced stimulation of glucose oxidation and partially inhibited the effect of NH+4. The mitochondrial enzymes pyruvate dehydrogenase and oxoglutarate dehydrogenase were activated by physiological concentrations of calcium in the isolated mitochondria. This effect was blocked by incubation with ruthenium red.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of a Rab3A effector domain-related peptide, CCK, and EGF in permeabilized pancreatic acini
Authors: Zeuzem S. , Stryjek-Kaminska D. , Caspary W. F. , Stein J. , Piiper A.
Published: December 2017

We report here that a synthetic peptide of the effector domain of the small-molecular-weight GTP-binding protein Rab3A (EDRab3AL) is a potent stimulator of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] production and amylase secretion in digitonin-permeabilized pancreatic acini. Moreover, the Rab3A effector domain peptide caused phosphatidylinositol 4,5-bisphosphate breakdown, indicating that the observed increase in Ins(1,4,5)P3 is due to stimulation of a phosphoinositide-specific phospholipase C (PLC). The dose-response curve for EDRab3AL-induced amylase release was biphasic, showing a maximum at 0.3 nM EDRab3AL and a decline at higher peptide concentrations. By contrast, the dose-response curve for EDRab3AL-induced Ins(1,4,5)P3 production was monophasic, showing stimulation with increasing EDRab3AL concentrations. A peptide of the effector domain of Rab1A, EDRab1AL, had no effect, indicating that the response to EDRab3AL is specific. Cholecystokinin octapeptide (CCK-8) and EDRab3AL had additive effects on the acinar Ins(1,4,5)P3 level. Epidermal growth factor (EGF), which has recently been shown to inhibit CCK-8-induced Ins(1,4,5)P3 production in pancreatic acinar cells, also decreased EDRab3AL-induced Ins(1,4,5)P3 production. These results suggest that EDRab3AL and CCK-8 act on the same EGF-inhibitable PLC by independent mechanisms. CCK-8 increased and EGF decreased amylase release in response to submaximal EDRab3AL concentrations. By contrast, at supramaximal EDRab3AL concentrations EGF increased and CCK-8 decreased EDRab3AL-stimulated amylase release. EDRab3AL had no effect in intact acini, indicating that the site of action of EDRab3AL is intracellular. We conclude that EDRab3AL regulates phosphoinositide-specific PLC activity and thereby amylase secretion in an analogous fashion to CCK-8, but from within the cell.(ABSTRACT TRUNCATED AT 250 WORDS)
Different modulation of hepatocellular Na+/H+ exchange activity by insulin and EGF
Authors: Haimovici J. , Beck J. S. , Molla-Hosseini C. , Vallerand D. , Haddad P.
Published: December 2017

Indirect evidence suggests that insulin, like epidermal growth factor (EGF), stimulates liver cell Na+/H+ exchange. We directly studied the effect of insulin on intracellular pH (pHi) and the Na+/H+ exchanger in isolated rat hepatocytes with the fluorescent probe, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). The effects of insulin were compared with those of EGF. Studies were carried out in the absence of HCO3- and in the presence of acetazolamide to isolate the Na+/H+ exchanger from other pH regulatory mechanisms. Insulin (9 nM) caused a reversible acidification of baseline pHi, whereas no significant effect was observed with EGF (30 nM). pHi was acidified by two different methods (NH4Cl pulse and external Na+ removal) to assess liver cell Na+/H+ exchange activity. In the NH4Cl pulse experiments, insulin had no significant effect on the Na+/H+ exchanger compared with the control (0.141 +/- 0.009 pH units/min, n = 14, and 0.122 +/- 0.023 pH units/min, n = 16, respectively). However, in the same conditions, EGF nearly doubled the rate of the Na+/H+ exchange activity (0.193 +/- 0.015 pH units/min, P < 0.05, n = 6). In the Na+ removal experiments, EGF again significantly increased the pHi recovery rate (0.542 +/- 0.032 pH units/min, n = 3) compared with the control (0.227 +/- 0.028 pH units/min, n = 5) and insulin (0.245 +/- 0.053 pH units/min, n = 5). Compared with control conditions, a subchronic administration of insulin (9 nM) in vitro had no significant effect on the Na+/H+ exchanger, nor did it affect baseline pHi.(ABSTRACT TRUNCATED AT 250 WORDS)