bluuebunny/crossref_metadata_2025 huggingface dataset:
Intracisternal TRH analogue RX 77368 stimulates gastric histamine release in rats
Authors: Yanagisawa K. , Tache Y.
Published: December 2017

The influence of intracisternal injection of the stable thyrotropin-releasing hormone (TRH) analogue RX 77368 on histamine levels in gastric secretion, interstitial fluid of the fundic submucosa, and portal hepatic circulation was investigated in rats. Intracisternal injection of RX 77368 (10-300 ng) induced a dose-related increase in histamine and acid output measured in the gastric secretion of pylorus-ligated, conscious rats. Intracisternal RX 77368 (300 ng) induced within 20 min a significant twofold histamine increase in interstitial fluid sampled from dialysis fibers implanted into the fundic submucosa. Histamine levels in the hepatic portal circulation were also dose dependently increased by RX 77368 injected intracisternally (30-100 ng), whereas intravenous infusion of RX 77368 (300 ng/30 min) did not significantly modify portal histamine levels. Bilateral cervical vagotomy or atropine pretreatment prevented intracisternal RX 77368-induced rise in hepatic portal levels of histamine, whereas purified gastrin monoclonal antibody 9303, injected at a dose blocking gastrin-stimulated acid secretion, had no effect. These results indicate that RX 77368 acts in the brain to increase gastric histamine secretion through vagal-dependent, muscarinic, nongastrin-mediated mechanisms and suggest a possible role of medullary TRH in the vagal regulation of gastric histamine secretion.
Effect of cellular iron concentration on iron uptake by hepatocytes
Authors: Trinder D. , Batey R. G. , Morgan E. H. , Baker E.
Published: December 2017

The effect of intracellular iron content on transferrin and iron uptake by cultured hepatocytes isolated from fetal rat liver was examined with ferric ammonium citrate and the iron chelator desferrioxamine (DFO). Incubation of the cells with ferric ammonium citrate for 24 h significantly increased the cellular nonheme iron level, whereas the number of transferrin binding sites and the uptake of transferrin and iron were reduced. In contrast, when iron-treated cells were incubated with DFO for 24 h, the cellular nonheme iron level was not altered, but the number of transferrin binding sites was increased. Treatment of the cells with exogenous iron and/or DFO did not affect the uptake of transferrin and iron by the nonsaturable processes. These results indicated that, in cultured hepatocytes, transferrin receptor expression and the subsequent uptake of transferrin and iron are regulated by the size of an intracellular, chelatable iron pool, whereas the uptake of iron by the nonsaturable processes is dependent on the extracellular transferrin concentration.
Comparison of calcium and magnesium absorption: in vivo and in vitro studies
Authors: Hardwick L. L. , Jones M. R. , Buddington R. K. , Clemens R. A. , Lee D. B.
Published: December 2017

Transport characteristics of Ca and Mg were compared at three different levels of the absorptive process in male (120-170 g) Wistar rats. Balance studies in intact rats revealed that fractional Ca absorption decreased with increased Ca intake so that net Ca absorption remained constant. Fractional Mg absorption decreased modestly with increased Mg intake so that net Mg absorption increased proportionately with increased dietary Mg. Everted duodenal sacs demonstrated the presence of active Ca absorption with serosal-to-mucosal (S/M) ratio of 2.65 +/- 0.20 (n = 6), which was greater than unity (P less than 0.001). In contrast, the S/M for Mg did not exceed unity. Mucosal duodenal Ca uptake exhibited a large saturable (Michaelis constant of 4.80 +/- 0.34 mM, maximal velocity of 4.71 +/- 0.13 nmol.min-1.mg-1) and a small nonsaturable component (0.12 +/- 0.01 nmol.min-1.mg-1) in 5-wk-old rats (120 g). In 72-wk-old rats (600 g) the diffusional component of Ca uptake became predominant, and the slope increased significantly to 0.32 +/- 0.01 (P less than 0.05). Duodenal Mg uptake was completely concentration dependent and exhibited no age-related changes.
Modulation of PAF-induced leukocyte adherence and increased microvascular permeability
Authors: Kubes P. , Suzuki M. , Granger D. N.
Published: December 2017

The objective of this study was to assess whether superoxide and leukocyte adhesion glycoproteins (CD18) mediate the leukocyte adherence to mesenteric microvessels and increased intestinal microvascular permeability induced by platelet-activating factor (PAF). PAF was infused into the arterial supply of an isolated autoperfused segment of cat intestine. Thirty minutes into the infusion, the number of adherent leukocytes within mesenteric venules was measured. This was followed by intravenous administration of either human recombinant superoxide dismutase (hSOD), hydrogen peroxide-inactivated hSOD, or a monoclonal antibody against the leukocyte adhesion molecule CD18 (MoAb IB4), and 30 min later, adherence measurements were repeated. hSOD and MoAb IB4 produced a 30 and 66% decrease, respectively, in leukocyte adherence, whereas inactivated hSOD had no effect. Adherence of PAF-activated cat neutrophils to plastic was reduced only by MoAb IB4, suggesting that PAF-induced leukocyte adherence is mediated by both CD18 and superoxide and that endothelium is necessary for the superoxide-mediated adhesion. In a correlate study, hSOD and MoAb IB4 were shown to attenuate the PAF-induced increase in microvascular permeability by 40 and 70%, respectively. These data indicate that the increased microvascular permeability induced by PAF can be attenuated when leukocyte adherence to microvascular endothelium is reduced using molecules that either bind to CD18 adhesive glycoproteins or scavenge superoxide.
Chicken gastrin: a member of the gastrin/CCK family with novel structure-activity relationships
Authors: Dimaline R. , Lee C. M.
Published: December 2017

The biological activity of natural chicken gastrins isolated from the "antrum"-like region of the chicken gut have been studied on gastric secretion in chickens, turkeys, and rats, pancreatic secretion in turkeys and rats, and gallbladder contraction in chickens and guinea pigs. Natural chicken gastrin was shown to be approximately 85% sulfated on the tyrosine that occurs at position 7 from the COOH-terminus. In both avian and mammalian systems, chicken gastrins were found to be potent stimulants of acid secretion but were virtually inactive as stimulants of pancreatic secretion and gallbladder contraction. Peptides with the COOH-terminal tetrapeptide amide of CCK and a sulfated tyrosine at position 7 from the COOH-terminus are usually potent stimulants of pancreas and gallbladder. However, although chicken gastrin has a CCK-like structure, it has a gastrin-like spectrum of biological actions. A proline immediately adjacent to the sulfated tyrosine may produce a steric effect that lowers the activity of chicken gastrin on pancreas and gallbladder. Evidently, then, the factors that determine specificity of action of CCK and gastrin are different in birds and mammals.
Capsaicin-sensitive afferent fibers contribute to gastric mucosal blood flow response to electrical vagal stimulation
Authors: Thiefin G. , Raybould H. E. , Leung F. W. , Tache Y. , Guth P. H.
Published: December 2017

Electrical stimulation of the peripheral vagus produces a noncholinergic increase in gastric mucosal blood flow (GMBF) via unknown mechanisms. The purpose of this study was 1) to investigate whether a portion of the increase in GMBF during prolonged electrical vagal stimulation involves a mechanism separate from augmented acid secretion and 2) to determine whether antidromic activation of afferent fibers contributes to the vascular or secretory responses to electrical vagal stimulation. Electrical vagal stimulation (40 V, 6 Hz, 2 ms) applied for 30 min to the distal cut end of the subdiaphragmatic ventral vagus significantly increased gastric acid secretion and GMBF measured by hydrogen gas clearance. Atropine (0.15 mg/kg iv) or omeprazole (10 mumol/kg iv) completely abolished the secretory response to electrical vagal stimulation, while a significant increase in GMBF remained. Pretreatment with perineural application of the sensory neurotoxin capsaicin to both cervical vagi significantly reduced by 48% the increase in GMBF but not gastric acid secretion; atropine completely abolished the remaining vascular response in capsaicin-treated rats. These results suggest that prolonged electrical vagal stimulation induces a sustained increase in GMBF partially independent of augmented acid secretion and that the noncholinergic portion of the vascular response is mediated by capsaicin-sensitive vagal afferent fibers.
Regulation of canine jejunal transit
Authors: Soper N. J. , Geisler K. L. , Sarr M. G. , Kelly K. A. , Zinsmeister A. R.
Published: December 2017

The aim of this study was to determine what factors influence the direction of movement of canine jejunal chyme. In four dogs, pacing electrodes were implanted near each end of a 50-cm jejunal Vella loop, while recording electrodes and intraluminal pressure catheters were spaced along the loop. After recovery, the loop was perfused from either the proximal stoma (forward flow) or the distal stoma (reversed flow), and effluent was collected from the nonperfused stoma. The pacesetter potentials were paced electrically in a forward (aborad) or a reverse (orad) direction. During control conditions (forward flow-forward pacing), the mean transit time of liquids was 2.6 +/- 0.1 min (mean +/- SE) and the static volume of the loop was 8.8 +/- 0.3 ml. Reversing both direction of flow and direction of pacesetter potential propagation slowed transit (4.4 +/- 0.4 min; P less than 0.05) and increased loop volume (16.0 +/- 1.0 ml; P less than 0.01). Reversing flow with forward pacing resulted in even slower transit (7.5 +/- 1.2 min; P less than 0.05), maintained a large volume (33 +/- 10 ml), and increased basal pressure in the loop from -1.2 +/- 1.7 (control) to 2.3 +/- 1.6 mmHg (P less than 0.05). These observations suggest that the direction of propagation of jejunal pacesetter potentials influences the direction of movement of jejunal chyme but that other factors have a role as well.
Identification of opioid receptors on gastric muscle cells by selective receptor protection
Authors: Grider J. R. , Makhlouf G. M.
Published: December 2017

Opioid receptors on isolated gastric smooth muscle cells were characterized pharmacologically by a technique in which synthetic selective opioid agonists and antagonists were used to protect and thus enrich a specific receptor type while all other receptors were inactivated by N-ethylamaleimide. Treatment of the cells with the selective mu-receptor agonist DAGO or antagonist CTAP preserved only the response to DAGO; treatment with the selective delta-receptor agonist DPDPE or antagonist naltrindole preserved only the response to DPPE; and treatment with the selective kappa-receptor agonist U50,488H or antagonist nor-binaltorphimine preserved only the response to U50,488H. The results established the presence of distinct kappa-, delta-, and mu-opioid receptors capable of mediating contraction of isolated gastric muscle cells. The pattern of interaction of endogenous opioid peptides with protected receptors implied that dynorphin-(1-13) and Met-enkephalin were selective agonists for kappa- and delta-opioid receptors, respectively, and Leu-enkephalin a preferential agonist of mu-opioid receptors. The results were confirmed by a reverse approach in which opioid receptors were inactivated by site-directed irreversible antagonists. beta-Funaltrexamine, a mu-selective antagonist, abolished the response to mu-receptor agonists, whereas beta-chlornaltrexamine, a mu- and kappa-selective antagonist, abolished the response to mu-receptor agonists and partially inhibited the response to kappa-receptor agonists.
Interconversions of lipid aggregates in rat and model bile
Authors: Little T. E. , Lee S. P. , Madani H. , Kaler E. W. , Chinn K.
Published: December 2017

The dynamic interchange of cholesterol and the phase transition between nonmicellar and micellar aggregates in rat and model bile were characterized with gel-permeation chromatography, quasi-elastic light scattering, turbidity measurements, and by radiolabeling lipid aggregates in bile. Cholesterol partitioned into either the micellar or nonmicellar phases independent of the lipid aggregate structure. In model bile, increasing bile salt concentrations led to a decrease in the relative proportion of nonmicellar aggregates beginning at 5 mM taurocholate (TC), while the relative cholesterol content of the nonmicellar fraction increased from 1.0 to 2.7 +/- 2.0 (means +/- SD). In rats, creation of a biliary fistula resulted in a decrease of bile salts from 41 to 4 mM. Mixed micelles increased from 25 to 120 A in radius, while nonmicellar aggregates increased from 180 to 800 A in radius. Addition of TC to model bile (cholesterol:lecithin = 1:1) vesicles with total lipid concentrations less than 7 mM yielded a progressive shift of vesicles (450 A) to mixed micelles (30 A). For mixtures with higher total lipid concentrations, addition of TC promoted substantial vesicle aggregation and resulted in formation of a third phase containing lipid aggregates larger in size than the initial vesicles. These results suggest that rapid exchange of cholesterol occurs in bile and that significant remodeling of vesicles can occur. These alterations in vesicles include both enrichment in cholesterol content and formation of larger aggregates during increases in bile salt concentration.
Characterization of a gastrin-type receptor on rabbit gastric parietal cells using L365,260 and L364,718
Authors: Roche S. , Bali J. P. , Galleyrand J. C. , Magous R.
Published: December 2017

Previous studies have demonstrated that gastrin and the COOH-terminal octapeptide of cholecystokinin (CCK-8) stimulated in vitro acid secretion from isolated rabbit gastric parietal cells. Both peptides bind to receptor sites located on these cells and induce an increase in phosphoinositide turnover and an uptake of [14C]aminopyrine ([14C]AP) with the same efficacy and potency. In the present study, we used the 3-(benzoylamino)-benzodiazepine analogue L365,260 and the 3-(acylamino)-benzodiazepine analogue L364,718 to investigate what type of receptor (gastrin type or CCK-A type) is involved in the regulation of the H+ secretory activity of the rabbit parietal cell. Neither L365,260 nor L364,718 alone caused stimulation of [3H]inositol phosphates ([3H]InsP) production. Each analogue inhibited 125I-labeled gastrin or 125I-CCK-8 binding to parietal cells and gastrin- or CCK-8-induced [3H]InsP production and [14C]AP accumulation. In all cases, L365,260 was approximately 70-100 times more potent than L364,718 (IC50 approximately 2-4 nM for L365,260 and approximately 0.2-0.4 microM for L364,718). Nevertheless, each antagonist displayed the same potency to inhibit the effects of gastrin or CCK-8. These results demonstrate that gastrin and CCK-8 interact with the same "gastrin-type" receptor on parietal cells. Moreover, L365,260 behaves as a competitive antagonist of the action of gastrin on parietal cells. Gastrin induces a rise in the levels of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P4] within the first seconds after parietal cell stimulation. The fact that L365,260 (10 nM) totally suppressed the gastrin-induced formation of Ins(1,4,5)P3 and Ins(1,3,4,5)P4 suggests the involvement of these isomers in the mediation of acid secretion through gastrin receptor activation.
Canine ileocolonic sphincter: flow, transit, and motility before and after sphincterotomy
Authors: Neri M. , Phillips S. F. , Fich A. , Haddad A. C.
Published: December 2017

Our hypothesis was that the canine ileocolonic sphincter (ICS), per se, would have little regulatory effect on the transit of chyme from ileum to colon. We argued, from earlier observations, that the ileocolonic junction was influenced more by functional motor integration of the ileum, ICS, and proximal colon. In five dogs, the ileocolonic sphincter was ablated by extramucosal sphincterotomy; the operation significantly lowered tonic pressures at the ICS. Animals were then studied in the fasting state and postprandially. Ileal flow was estimated by marker dilution, and ileocolonic flow was estimated by total recovery of chyme from a colonic cannula. Transit times were measured after the bolus infusion of nonabsorbable markers. Ileocolonic sphincterotomy did not significantly alter flow rates or transit times of chyme across the ICS, although ileal motor patterns were changed after sphincterotomy. We concluded that the ICS probably has only a small effect on transit and flow at the ileocolonic junction. These findings argue for the importance of integrated motility of the ileum, ICS, and proximal colon in controlling the flow of chyme in this region.
PAF increases hepatic vascular resistance and glycogenolysis in vivo
Authors: Hines K. L. , Braillon A. , Fisher R. A.
Published: December 2017

Administration of platelet-activating factor (PAF) to portal venous circulation of anesthetized fed rats produced decreases in mean arterial pressure and increases in hepatic portal pressure and blood glucose concentration. These responses to PAF were dose dependent with ED50 values of 0.02-0.05 micrograms/kg and specific as lyso- and enantio-PAF did not reproduce effects of PAF. Specific PAF receptor antagonist SRI 63-675 (75 micrograms/kg) inhibited significantly these PAF (0.1 micrograms/kg)-induced responses in rats. Administration of prazosin (500 micrograms/kg) and propranolol (400 micrograms/kg) to rats abolished phenylephrine (50 micrograms/kg)-induced increases in mean arterial pressure, hepatic portal pressure, and blood glucose concentration but did not prevent PAF (1 microgram/kg)-induced alterations in these parameters. Glycogen phosphorylase alpha levels were increased significantly in livers of rats after administration of PAF (1 microgram/kg) or phenylephrine (50 micrograms/kg). Administration of prazosin and propranolol to rats inhibited phenylephrine- but not PAF-induced activation of hepatic glycogen phosphorylase. Hepatic adenosine 3',5'-cyclic monophosphate (cAMP) concentration was increased slightly by PAF, but these increases were eliminated by adrenergic blockade, suggesting that activation of hepatic glycogen phosphorylase by PAF is not dependent on increases in cAMP. Increases in hepatic portal pressure and blood glucose concentration in response to PAF (0.1 micrograms/kg) were not attenuated in adrenalectomized rats. Moreover, PAF (0.1 micrograms/kg) stimulated increases in hepatic portal pressure after administration of the ganglionic blocking agent chlorisondamine (2.5 mg/kg) to adrenalectomized rats. Administration of PAF (0.05 micrograms/kg) to rats instrumented with pulse Doppler flow probes produced decreases in hepatic arterial and portal vein blood flow and increases in hepatic arterial and portal vascular resistance. These observations provide direct evidence that PAF regulates hepatic hemodynamics and glycogenolysis in vivo. It is suggested that PAF plays an important role in regulating hepatic blood flow and supplying extrahepatic tissues with energy substrates by sympathetic-independent mechanism(s) after its release in acute pathophysiological situations.
Endotoxin-induced increases in regional glucose utilization by small intestine: a TNF-independent effect
Authors: Lang C. H. , Obih J. A. , Bagby G. J. , Bagwell J. N. , Spitzer J. J.
Published: December 2017

The purpose of the present study was to determine whether the lipopolysaccharide (LPS)-induced increase in glucose uptake was a generalized response along the length of the entire gastrointestinal (GI) tract and to assess the relative contributions of the mucosa and muscularis. The putative roles of tumor necrosis factor (TNF) and blood flow in the metabolic response of the intestine to LPS were also examined. In vivo glucose uptake (Rg) was determined for various segments of the GI tract under basal postabsorptive conditions and 3 h after intravenous injection of Escherichia coli LPS using 2-[14C]deoxyglucose. At this time, LPS-treated rats were euglycemic, and Rg was elevated in all sections of the GI tract (52-96%). In control animals, mucosal Rg accounted for 79% of the glucose uptake by the entire small intestine; LPS increased Rg in both the mucosa and muscularis and did not alter this relationship. The LPS-induced increase in intestinal Rg was not attenuated by pretreatment with TNF antibody. Cardiac output (CO) and intestinal blood flow, assessed using radiolabeled microspheres, were not different from control values 3 h after LPS. Blood flow to the muscularis was increased (120%) in all sections of the small intestine from LPS-treated rats. These results indicate that 3 h after a low dose of LPS, glucose uptake by the entire length of the GI tract was elevated, and the majority of increase was due to enhanced uptake by the mucosa, which was blood flow independent. Furthermore, the increased Rg was not dependent on elevations in plasma glucose, insulin, or TNF levels. However, an increased blood flow to the muscularis was associated with an elevated Rg in that region.
Effect of exercise on intestinal motility and transit in trained athletes
Authors: Soffer E. E. , Summers R. W. , Gisolfi C.
Published: December 2017

We measured the effect of exercise, at different intensities, on duodenojejunal motor activity in eight trained, ambulant cyclists, using a catheter with two strain gauge transducers connected to a solid-state data logger. Orocecal transit time was measured by H2 breath test. Subjects were studied for two consecutive days and fed twice a day with a similar, 860-kcal meal. Control recordings on the first day were made at rest and a naloxone infusion was given after the evening meal. On the second day, the athletes exercised after meals for 20 min at 80% of peak O2 uptake (VO2max) and naloxone was infused during the second session. On separate days, five subjects exercised for 3 h at 60% VO2max and for 10 min at 90% VO2max. Plasma concentrations of beta-endorphin were determined before and after exercise. The fed pattern was interrupted by an activity front during 2 of 16 sessions of exercise at 80% VO2max, in the presence of naloxone, and 5 of 8 sessions at 90% VO2max, with and without naloxone. It was not affected by exercise at 60% VO2max. Orocecal transit was not affected by intense exercise. The results show that exercise can affect intestinal postprandial motor activity. The effect is intensity related and may not be mediated by opioids.
Functional specificity of jejunal brush-border pteroylpolyglutamate hydrolase in pig
Authors: Chandler C. J. , Harrison D. A. , Buffington C. A. , Santiago N. A. , Halsted C. H.
Published: December 2017

To determine the functional specificity of intestinal brush-border pteroylpolyglutamate hydrolase (PPH), we compared the regional location of in vivo hydrolysis of pteroyltriglutamate (PteGlu3) with the location of activity and immunoreactivity of the enzyme in the pig. After in vivo incubations, PteGlu3 hydrolytic products were recovered from intestinal segments in the jejunum but not from the ileum. Brush-border PPH activity in fractionated mucosa was 10-fold greater in the jejunum than in the ileum, whereas the activity of intracellular PPH was increased in the distal ileum. Antibodies to purified brush-border PPH identified a major protein band at 120 kDa and a minor protein band at 195 kDa in solubilized jejunal brush border. Immunohistochemistry identified the enzyme only on the brush-border surface of the jejunum, whereas an immunoblot of solubilized brush-border membranes identified brush-border PPH in the jejunum but not in the ileum. The parallel of the regional location of in vivo hydrolysis of PteGlu3 with the location of brush-border PPH activity and immunoreactivity demonstrates the functional specificity of this enzyme in folate digestion.
Myosin phosphorylation and calcium in tonic and phasic contractions of colonic smooth muscle
Authors: Gerthoffer W. T. , Murphey K. A. , Mangini J. , Boman S. , Lattanzio F. A.
Published: December 2017

The time dependence of lightly loaded shortening velocity, myosin phosphorylation, and changes in myoplasmic Ca2+ concentration ([Ca2+]i) were measured during tonic and phasic contractions of circular smooth muscle from the proximal colon of the dog. Shortening velocity was measured by quick release to a 10% afterload. Myosin phosphorylation was measured by an immunoblot method, and changes in [Ca2+]i were estimated by measuring fluorescence intensity at 550 nm in muscle strips loaded with fluo-3. During tonic contractions induced by 60 mM K+, phosphorylation increased monotonically from 0.11 +/- 0.011 to 0.29 +/- 0.015 mol Pi/mol light chain at 10 min. In contrast, lightly loaded shortening velocity increased rapidly within 10 s to 0.042 +/- 0.003 lengths/s and decreased exponentially to 0.013 +/- 0.001 lengths/s at 15 min. During transient contractions induced by 100 microM acetylcholine, phosphorylation increased from 0.16 +/- 0.03 to 0.30 +/- 0.06 mol Pi/mol light chain at 19 s. In contrast, shortening velocity increased to 0.068 +/- 0.015 lengths/s within 2.4 s and decreased significantly to 0.027 +/- 0.009 lengths/s at 22 s. Fluo-3 fluorescence increased in parallel with force during both tonic and transient contractions. In a smooth muscle that is able to contract both tonically and phasically we observed transient increases in shortening velocity without concurrent phosphorylation or [Ca2+]i transients. Therefore, there are factors in addition to myosin phosphorylation or changes in [Ca2+]i that regulate cross-bridge cycling rates in both tonic and phasic contractions.
Regulation of intestinal cholecystokinin and somatostatin mRNA by bombesin in rats
Authors: Kanayama S. , Liddle R. A.
Published: December 2017

The neuropeptide bombesin has been shown to stimulate secretion of several gastrointestinal hormones, including cholecystokinin (CCK). We have previously demonstrated that stimulation of CCK release by feeding is associated with an increase in steady-state intestinal CCK mRNA levels. The purpose of the present study was to determine whether bombesin stimulates CCK release in rats and, if so, to determine whether bombesin regulates CCK mRNA levels in a manner similar to that of feeding. To establish a proper dose of bombesin for stimulating CCK release, rats received 1-h intravenous infusions of 0.25, 1, 4, or 16 micrograms.kg-1.h-1 bombesin. Basal plasma CCK levels averaged 1.8 +/- 0.4 pM and increased to peak levels of 2.9 +/- 0.6 pM within 15 min of infusion with 4 micrograms.kg-1.h-1 bombesin (the maximally effective dose). With the use of this dose, rats then received infusions of bombesin or saline lasting up to 24 h. At 1, 2, 4, and 24 h, animals were killed for collection of plasma for CCK measurements and of intestine for measurements of intestinal CCK and somatostatin mRNA levels. Bombesin treatment stimulated an increase in plasma CCK levels at 1 h, but levels declined to basal by 4 h, where they remained at 24 h. Despite increasing plasma CCK levels, bombesin infusion, unlike dietary stimulation, had no effect on duodenal CCK mRNA levels. Finally, to determine whether the decrease in plasma CCK levels after prolonged bombesin treatment was due to tachyphylaxis, rats treated with bombesin for 4 h were also fed soybean trypsin inhibitor (a known stimulus of CCK secretion).(ABSTRACT TRUNCATED AT 250 WORDS)
Stimulatory and inhibitory actions of carbachol on chloride secretory responses in human colonic cell line T84
Authors: Warhurst G. , Higgs N. B. , Tonge A. , Turnberg L. A.
Published: December 2017

The muscarinic agonist carbachol (CCh) was shown to elicit both stimulatory and inhibitory actions on Cl- secretion in T84 cells. These effects were observed in cells that had been prestimulated with adenosine 3',5'-cyclic monophosphate (cAMP) agonists. The addition of CCh to cells treated with submaximal concentrations (1-10 nM) of the receptor-mediated agonist prostaglandin (PG) E2 resulted in a biphasic effect on short-circuit current (Isc) involving a transient synergistic rise followed by a slower and sustained attenuation of the PGE2-activated Isc response. In contrast at higher PGE2 concentrations (greater than 10 nM) or with nonreceptor-mediated cAMP agonists (forskolin, dibutyryl cAMP) CCh elicited a prolonged synergistic response. Both effects of CCh could be reproduced by selective activation of the Ca2+ pathway. Increasing cytosolic Ca2+ with ionophore A23187 partially mimicked the "early" stimulation of secretion, though there was evidence that a combination of A23187 and the protein kinase C activator phorbol 12,13-dibutyrate (PDB) was required for full expression of the secretory response. In contrast, treatment with PDB alone closely mimicked the CCh-induced inhibition of PGE2-stimulated Isc. The antisecretory effects of CCh were associated with a marked attenuation of cAMP production in response to receptor-mediated agonists, including PGE2 and vasoactive intestinal peptide. This effect could also be closely mimicked by PDB. Pretreatment with pertussis toxin partially inhibited the ability of CCh to inhibit cAMP production. CCh potentiated the cAMP response to the nonreceptor agonist forskolin.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of Pluronic L81, a hydrophobic surfactant, on intestinal mucosal cholesterol homeostasis
Authors: Pool C. , Nutting D. F. , Simmonds W. J. , Tso P.
Published: December 2017

Addition of triglyceride and phospholipid to sodium taurocholate when chylomicron output was blocked by L81 did not increase lymphatic total cholesterol output or mucosal unesterified (UC) content more than with sodium taurocholate alone, but mucosal esterified cholesterol (CE) was increased slightly. In these animals with defective chylomicron formation, excess cholesterol accumulated in the intestinal mucosa mainly as CE. The mucosal cholesterol content of animals with normal chylomicron transport expanded during cholesterol and triglyceride absorption, and the expansion led to increased lymphatic secretion of CE. These animals accumulated significantly less CE in their mucosa than did rats treated with L81, but had about the same amount of mucosal UC. However, the overall uptake of cholesterol from the lumen, as determined by either radioactivity or mass of cholesterol in mucosa and lymph, was significantly less in the L81 rats. Also, more radioactive cholesterol remained in the lumen of the L81-treated rats. Finally, the data on specific activities of free and esterified cholesterol showed that the mucosal cholesterol derived from the lumen does not mix evenly with the free cholesterol pool in the enterocytes and is preferentially esterified for export in lymph as triglyceride-rich lipoprotein.
Electric properties of rat liver cell cultures on gas-permeable membranes
Authors: Wehner F. , Guth D.
Published: December 2017

In rat hepatocytes grown on gas-permeable membranes (Petzinger et al. In Vitro Cell. Dev. Biol. 24: 491-499, 1988), cellular and canalicular potentials as well as input resistances were measured using two-channel microelectrodes. In HCO3(-)-containing solutions, we found -30.9 +/- 0.4 (SE) (n = 141) and -13.9 +/- 1.4 mV (n = 22) for cell and canalicular membrane potentials, respectively. There was no dependence of these parameters on the age of the primary culture. Canalicular input resistance, however, increased from 13.3 +/- 2.0 M omega (n = 4) at day 1 after seeding to 36.1 +/- 5.0 M omega (n = 9) at day 2 and stabilized thereafter, while cell input resistance continuously decreased from 37.0 +/- 3.3 M omega at 1 h (n = 6) to 5.2 +/- 2.1 M omega (n = 27) at 3 days after preparation. In ion substitution experiments there were no changes in the transference numbers for K+, Na+, or Cl- that could account for this effect. Cable analysis, however, revealed that the decrease in input resistance reflects a time-dependent increase in electrical coupling between cells. We conclude that rat liver cells on gas-permeable membranes are highly suited for the quantitative analysis of cell-to-cell interaction. In addition, cells and canaliculi are readily accessible with two-channel microelectrodes, making this preparation a promising tool for electrophysiological analysis of hepatocellular transport mechanisms.
Comparison of fluid absorption by bovine and ovine descending colon in vitro
Authors: McKie A. T. , Goecke I. A. , Naftalin R. J.
Published: December 2017

Although bovine and ovine descending colon absorbed solute and sodium at approximately the same rate in vitro, water absorption by bovine colon was faster (15.0 +/- 2.0 microliters.h-1.cm-2, n = 8) than by ovine colon (8.6 +/- 1.3 microliters.h-1.cm-2, n = 9; P less than 0.01). Consequently, the observed osmolality of cattle absorbate was lower (364 +/- 13 mosmol/kg, n = 8) than with sheep (807 +/- 135 mosmol/kg, n = 9; P less than 0.01). Paracellular permselectivity was examined to elucidate this difference; the permeability of bovine descending colon to [3H]polyethylene glycol 400 and 4000 was higher than in sheep (P less than 0.001). A paracellular solvent drag was observed in bovine but not in ovine colon, and the electrical resistance of bovine colon was lower (16.0 +/- 1.4 omega.cm2) than ovine colon (28.0 +/- 2.4 omega.cm2; P less than 0.001). Pore radii of 2.5 nm for ovine and 5 nm for bovine colonic paracellular route were estimated from these data. It is concluded that an increased hydraulic conductance of the "active" route for solute absorption combined with raised solute reflux via the wider paracellular pathway may account for the failure of cattle to form a hypertonic absorbate and, consequently, hard feces.
Stimulation of proximal small intestinal mucosal growth by luminal polyamines
Authors: Wang J. Y. , McCormack S. A. , Viar M. J. , Johnson L. R.
Published: December 2017

The purpose of this study was to determine whether luminal polyamines stimulate intestinal mucosal growth in vivo. Rats received 2% alpha-difluoromethylornithine (DFMO) added to their drinking water throughout the experiment. The polyamines spermidine and spermine (3 mg each/100 g body wt) were given intragastrically in combined doses once at 9:30 A.M. and again at 5:30 P.M. Duodenal and jejunal mucosal ornithine decarboxylase (ODC) activity in the DFMO-treated rats was inhibited significantly for the duration of the study. DFMO also markedly decreased the rate of [3H]thymidine incorporation into DNA of duodenal and jejunal mucosa. The decrease in [3H]thymidine incorporation was significant 4 days and maximal 6 and 8 days after beginning treatment with DFMO. Decreased ODC activity and DNA synthesis were paralleled by decreases in total mucosal DNA, RNA, and protein content. Administration of the polyamines significantly reversed the effects of DFMO except the inhibition of ODC. In fact, there were no significant differences in mucosal growth parameters between the controls (without DFMO) and those treated with DFMO plus polyamines. Oral administration of spermidine and spermine at a dose of 4.5 mg each/100 g body wt for 6 days to rats not treated with DFMO increased the normal rate of mucosal growth in the duodenum and jejunum as well. Polyamine accumulation in IEC-6 cells was measured to determine whether it was altered by DFMO. IEC-6 cells took up [3H]putrescine and [3H]spermidine from their surrounding environment and the uptake was stimulated by serum. DFMO (5 mM) totally inhibited the increase in ODC activity but had no effect on the cellular uptake of polyamines in the presence of putrescine.(ABSTRACT TRUNCATED AT 250 WORDS)
Ornithine decarboxylase is involved in repair of small intestine after ischemia-reperfusion in rats
Authors: Fujimoto K. , Granger D. N. , Price V. H. , Tso P.
Published: December 2017

To assess whether ornithine decarboxylase (ODC) is involved in mucosal repair after ischemia-reperfusion (I/R), two approaches were used: 1) measurement of mucosal ODC activity at different intervals after I/R, and 2) inhibition of ODC activity with alpha-difluoromethylornithine (DFMO, an irreversible inhibitor). In the first series of experiments, rats were allowed to recover after superior mesenteric arterial occlusion (10 min) before harvesting the intestinal mucosa for measurement of ODC activity. ODC activity increased markedly 6 h after I/R, and greater than 72 h were required for enzyme activity to return to normal. DFMO treatment completely abolished the I/R-induced increase in ODC activity. In another series of experiments, rats with an intestinal lymph fistula were infused intraduodenally at 3 ml/h with vehicle or 2% DFMO in vehicle immediately after I/R. Lipid absorption was measured at 24 and 48 h after I/R. In the DFMO group, lymph radioactive lipid output at 24 h after I/R was significantly lower compared with time-matched sham-operated controls. Lymph lipid output in rats receiving the vehicle was restored to a normal level at 48 h after I/R. However, this restoration of normal lymphatic lipid transport at 48 h I/R was not observed in the DFMO group. These observations indicate that ODC activity plays an important role in the repair process that results in complete restoration of mucosal function 2 days after I/R.
In vivo measurement of rat gastric surface cell intracellular pH
Authors: Kaneko K. , Guth P. H. , Kaunitz J. D.
Published: December 2017

Intracellular pH (pHi) of the surface mucous cells of the rat stomach was measured in vivo using a fluorescent microscopic technique. Gastric surface mucous cells were loaded with fluorescein by superfusion with 5(6)-carboxy-fluorescein diacetate, and the ratio of the intensity of emitted fluorescence at 495 and 450 nm excitation was determined by image analysis of videotapes. An in vivo calibration curve was constructed using 10(-5) M nigericin to equilibrate pHi and extracellular pH, and intra- and extracellular potassium concentrations. The pHi of the gastric surface cells was 7.22 +/- 0.06, and pHi was stable over 45 min. A pulse of 20 mM NH4Cl in Krebs solution rapidly alkalinized the cells to pH 7.7, followed by a slow return of pHi to baseline pH 7.2. After withdrawal of NH4Cl, pHi dropped to 6.7 with gradual recovery to baseline. pHi increased when the mucosa was superfused, in the presence of nigericin, with 150 mM KCl solutions and decreased when superfused with 2.6 mM KCl solutions. Perfusing with pH 3 Krebs buffers transiently decreased pHi, whereas superfusion with pH 1.2 Krebs irreversibly decreased pHi. These findings confirm that the measured fluorescence was intracellular and that the cells responded to changes in luminal pH.
Variation of slow-wave frequency and locking during the migrating myoelectric complex in dogs
Authors: Caenepeel P. , Janssens W. , Accarino A. , Janssens J. , Vantrappen G. , Eyssen H.
Published: December 2017

Slow waves determine rhythm and polarity of spike bursts. We measured the variation of slow-wave frequency (swf) and locking (swl) in the canine jejunum during the various phases of the migrating myoelectric complex (MMC) and during induced phase III (erythromycin 125 micrograms/kg iv bolus or somatostatin 2.5 micrograms.kg-1.h-1 iv infusion), blocked phase III (atropine 20 micrograms/kg iv bolus), and so-called stationary phase III activity (cisapride 150 micrograms/kg iv bolus). The EMG of 4 dogs, implanted with 10 bipolar electrodes, was recorded on a polygraph. Our results indicate that swf and swl change during the MMC from a stepwise swf gradient with slow waves locked in plateaus during phase I to a continuous swf gradient without or with significantly reduced phase locking during phase III. The length of the first swf plateau decreases significantly from 42 +/- 12 cm post Treitz during phase I to 11 +/- 4 cm during spontaneous phase III. Atropine block of phase III activity prevents phase unlocking and development of a continuous swf gradient. Our hypothesis is that phase unlocking may be one of the induction mechanisms of spike-burst activity.
Dietary calcium phosphate inhibits cytotoxicity of fecal water
Authors: Lapre J. A. , De Vries H. T. , van der Meer R.
Published: December 2017

The effects of dietary steroid and CaHPO4 supplementation on the solubility of bile acids and cytotoxicity of fecal water were studied in rats. Dietary steroid supplementation increased the bile acid concentration of both feces and fecal water. CaHPO4 supplementation produced a slight increase in total fecal bile acid concentration but resulted in a drastic decrease in soluble bile acid concentration. Cytotoxicity of fecal water on control and steroid-supplemented diets decreased with CaHPO4 supplementation analogous to the decrease in soluble bile acid concentration. The concentrations of precipitated Ca and Pi were highly correlated (r greater than 0.90) with the concentration of precipitated bile acids and with inhibition of cytotoxicity. However, there was no significant correlation between the logarithms of soluble calcium and soluble bile acids, indicating that solubility of bile acids is not determined by soluble calcium. Concentrations of calcium and phosphate in fecal water indicated the formation of insoluble calcium phosphate in the intestine. Thus dietary CaHPO4 causes a decrease in soluble bile acid concentration, which is probably due to the formation of an insoluble bile acid calcium phosphate complex. Consequently, cytotoxicity of fecal water is inhibited, which might have implications for the protective effect of dietary calcium with regard to colonic cancer in humans.
CRF in the paraventricular nucleus mediates gastric and colonic motor response to restraint stress
Authors: Monnikes H. , Schmidt B. G. , Raybould H. E. , Tache Y.
Published: December 2017

The role of corticotropin-releasing factor (CRF) in the paraventricular nucleus of the hypothalamus (PVN) in the control of gastric emptying of a nonnutrient meal and colonic transit was investigated in conscious fasted rats chronically implanted with hypothalamic cannulas and catheters in both the stomach and proximal colon. CRF (0.06-0.6 nmol) microinfused unilaterally into the PVN resulted in a dose-dependent inhibition of gastric emptying (0-51%) and stimulation of colonic transit (0-93%). CRF (0.6 nmol)-induced delay in gastric emptying was inhibited by 50% by subdiaphragmatic vagotomy or atropine methyl nitrate (1 mg/kg ip), whereas the stimulation of colonic transit was completely abolished by atropine methyl nitrate and reduced by 19% by vagotomy. Microinfusion of CRF (0.6 nmol) into the lateral hypothalamus or central amygdala had no effect. Restraint exposure for 1 h delayed gastric emptying and stimulated colonic transit by 28 and 78%, respectively. Bilateral microinfusion of the CRF antagonist alpha-helical CRF-(9-41) (13 nmol) into the PVN before restraint abolished stress-induced alterations of gastric and colonic transit. The CRF antagonist did not alter basal gastric and colonic transit under basal conditions. These data indicate that the PVN is a specific responsive site for central CRF-induced alterations of gastric and colonic transit and suggest that endogenous CRF in the PVN plays a role in mediating restraint stress-related alterations of gastrointestinal transit.
NaCl-dependent expression of amiloride-blockable Na+ channel in Xenopus oocytes
Authors: Asher C. , Singer D. , Eren R. , Yeger O. , Dascal N. , Garty H.
Published: December 2017

RNA was isolated from chicken lower intestine (both colon and coprodeum) and injected into Xenopus oocytes. 22Na+ fluxes measured after 1-4 days demonstrated the induction of an amiloride-blockable pathway. The Na+ transporter expressed by the exogenous RNA had a high affinity to amiloride (inhibitory constant less than 0.1 microM), but was insensitive to ethylisopropyl amiloride, i.e., it is likely to be the apical Na+ channel. Functional channels were readily expressed in oocytes injected with RNA derived from chickens fed a low-NaCl diet. On the other hand, no channel activity was detected in oocytes injected with RNA isolated from chickens fed a high-NaCl diet. Thus the previously reported regulation of transport by the dietary NaCl intake involves modulations in the level of mRNA that codes either for the Na+ channel or a posttranscriptional regulator of the channel.
Bile secretion of sulfated glycolithocholic acid is required for its cholestatic action in rats
Authors: Kuipers F. , Hardonk M. J. , Vonk R. J. , van der Meer R.
Published: December 2017

To test our hypothesis that the cholestatic action of sulfated glycolithocholic acid (SGLC) in the rat is related to its interaction with calcium in the biliary tree [R. van der Meer, R. J. Vonk, and F. Kuipers. Am. J. Physiol. 254 (Gastrointest. Liver Physiol. 17): G644-G649, 1988], we have now compared its effects on bile formation in control Wistar rats and mutant Groningen Yellow (GY) Wistar rats. Intravenous injection of 0.6 mumol/100 g body wt of [14C]SGLC in unanesthetized rats with permanent biliary drainage did not induce cholestasis in either of the strains; however, its biliary secretion was strongly impaired in GY rats (12% dose at 1 h after injection vs. 95% dose in controls). Injection of 6.0 and 12.0 mumol/100 g body wt of [14C]SGLC caused an almost complete cessation of bile flow in control rats within 3 and 1 h, respectively. In contrast, administration of the same doses did not cause cholestasis in GY rats. Cholestasis in control rats was preceded by coprecipitation of [14C]SGLC and calcium in bile and incomplete biliary recovery of radioactivity. The hepatic content 15 min after injection of [14C]SGLC (6.0 mumol/100 g body wt) was similar in control and GY rats, 51 and 49% of the dose, respectively. Administration of glycolithocholic acid, the unsulfated parent compound of SGLC (6.0 mumol/100 g body wt), induced a rapid but reversible cessation of bile flow in both controls and GY rats; in this case no precipitation was observed in bile. This study shows that rapid bile secretion of SGLC is required for the induction of cholestasis.(ABSTRACT TRUNCATED AT 250 WORDS)
Yersinia enterocolitica enteritis affects rabbit intestinal longitudinal smooth muscle function
Authors: Scott R. B. , Tan D. T.
Published: December 2017

To determine whether Yersinia enterocolitica (YE) enteritis has an effect on the biomechanical properties of intestinal smooth muscle, New Zealand White rabbits (600-900 g) were divided into an infected group (n = 9) and sham-infected animals fed ad libitum (n = 9), or pair fed with the infected group (n = 9). Animals were inoculated with 10(10) organisms of YE in 10 ml NaHCO3 (infected group) or 10 ml NaHCO3 (sham-infected control and pair-fed groups) at time 0. Daily food intake, weight gain, and YE excretion were noted. Six days later animals were killed and longitudinal smooth muscle strips prepared from proximal (P), medial (M), and distal (D) segments of intestine in each treatment group. Isometric tension was recorded in tissue baths perfused with oxygenated Krebs solution and 10(-6) M tetrodotoxin. Basal and active (the response to 10(-5) M carbachol) length-tension curves were generated. Then, with the muscle strips stretched to their optimum length for tension development, the dose response to carbachol and to graded depolarization with KCl was determined. Infected animals had a significantly reduced food intake and weight gain compared with controls. The development of basal tension with stretch was not significantly different in infected compared with control or pair-fed tissues from the same site.(ABSTRACT TRUNCATED AT 250 WORDS)