bluuebunny/crossref_metadata_2025 huggingface dataset:
Electrical and mechanical interactions between the muscle layers of canine proximal colon
Authors: Sabourin P. J. , Kingma Y. J. , Bowes K. L.
Published: December 2017

Electrical and mechanical interactions between the two smooth muscle layers of canine colon have been studied using a dual sucrose gap apparatus. Muscle samples were dissected into an L-shape, with one leg cut in the circular direction and the other cut in the longitudinal direction. Longitudinal muscle was removed from the circular leg and circular muscle was removed from the longitudinal leg. The bend of the L contained both layers. The activity of the two layers was studied simultaneously under basal conditions, after stimulation by neostigmine and carbachol, and in the presence of tetrodotoxin. Interactions were more common after stimulation and were marked by modification of one layer's mechanical and electrical activity during increased activity in the other layer. Two patterns were commonly observed. First, during a burst of membrane potential oscillations and spike potentials in the longitudinal layer, slow waves in the circular layer developed spike potentials and some slow waves were also prolonged. Second, during a slow-wave cycle in the circular layer, the amplitude of membrane potential oscillations in the longitudinal layer was increased with an associated increase in the incidence of spike potentials. These interactions were associated with contractions of increased strength, which were similar in both layers. All interactions continued after nerve-conduction blockade by tetrodotoxin.
Glutathione disulfide as index of oxidant stress in rat liver during hypoxia
Authors: Jaeschke H.
Published: December 2017

Formation of glutathione disulfide (GSSG) was used as an index of reactive oxygen generation in the isolated perfused liver of male Fischer rats during normoxia and hypoxia. Low oxygen tension may affect GSSG formation, rereduction, and transport mechanisms. The effect of short-term hypoxia (15 min) on the biliary and sinusoidal transport of GSSG was tested with the glutathione S-conjugates of sulfobromophthalein and 1-chloro-2,4-dinitrobenzene. Hypoxia inhibited S-conjugate excretion through both pathways by 15-20%. tert-Butyl hydroperoxide (75 microM tBHP) or diquat (200 microM) in the perfusate increased hepatic GSSG release by 430 and 1,550%, respectively, and increased the tissue GSSG content by 47 and 124%, respectively, under normoxia. Hypoxia reduced the stimulated GSSG export by 38 (tBHP) and 83% (diquat) and also caused an additional increase of the tissue GSSG content by 112% during tBHP infusion but caused a reduction by 32% during diquat infusion. Inhibition of the biliary export of GSSG and S-conjugates is mainly compensated by the sinusoidal efflux. Therefore, it is concluded that hypoxia reduces GSSG formation predominantly through suppression of reactive oxygen formation with only marginal effects on the biliary and sinusoidal excretion mechanism. Thus hepatic GSSG formation is a sensitive indicator of oxidant stress during normoxia and hypoxia. Because single parameters may vary considerably, simultaneous monitoring of GSSG in bile, perfusate, and tissue is essential for qualitative and quantitative estimation of reactive oxygen formation.
Iron uptake from transferrin and lactoferrin by rat intestinal brush-border membrane vesicles
Authors: Kawakami H. , Dosako S. , Lonnerdal B.
Published: December 2017

Interaction of 59Fe-labeled rat transferrin, human lactoferrin, and bovine lactoferrin with rat small intestinal brush-border membrane vesicles was investigated with the use of a rapid filtration technique. Specific binding of 59Fe-labeled rat transferrin and bovine lactoferrin to brush-border membrane vesicles from suckling and adult rats was identified. In contrast, no binding of human lactoferrin occurred. The presence of transferrin receptors on the brush-border membrane of suckling rats was confirmed by immunoblotting, and the molecular mass of the receptor was 96 kDa under nonreducing conditions. Scatchard plot analysis indicated 2.4 x 10(14) binding sites/mg of membrane protein with an affinity constant (Ka) of 4.9 x 10(6) M-1 for rat milk transferrin and 2.2 x 10(14) bi
Development of gastric antral smooth muscle contractility in newborn rabbits
Authors: Zitterman J. , Ryan J. P.
Published: December 2017

Muscle strips from newborn and adult rabbits were stimulated with acetylcholine (ACh; receptor activation), potassium (membrane depolarization), and calcium (skinned muscle preparation). The study was designed to determine whether postnatal development is associated with agonist-specific increases in force development or with an overall increase in antral smooth muscle contractility. The results can be summarized as follows. 1) Antral smooth muscle from newborn animals developed less active force than tissues from adult animals when stimulated with ACh. 2) Antral smooth muscle from newborn animals developed less active force than tissues from adult animals when stimulated with potassium. 3) Skinned muscle preparations from newborn animals developed less active force than skinned muscle fibers from adult animals when stimulated with calcium. The data suggest that the overall contractility of antral smooth muscle from the newborn is less than that of the adult and that the difference can be attributed, at least in part, to factors past the level of the membrane. The study does not rule out the possibility that differences might also exist with respect to the availability or mobilization of calcium.
Medullary raphe: a new site for vagally mediated stimulation of gastric motility in cats
Authors: Hornby P. J. , Rossiter C. D. , White R. L. , Norman W. P. , Kuhn D. H. , Gillis R. A.
Published: December 2017

Thyrotropin-releasing hormone (TRH) is clearly implicated in the control of gastric function via interactions in the dorsal motor nucleus of the vagus (DMV) of the cat. The source of the TRH innervation of the DMV is important to determine because this region could be of importance in control of gastric function. TRH-immunoreactive (ir) neurons are located in the raphe obscurus (Ro), raphe pallidus (Rp), and raphe magnus (Rm). Retrograde tracer applied to the DMV resulted in the most numerous labeled neurons in the caudal Ro and Rp in the same region where TRH-ir neurons are located. To address the question whether DMV-projecting neurons in the raphe subnuclei play a role in control of gastric motility, the following experiments were performed in alpha-chloralose-anesthetized cats while recording pyloric motility and blood pressure. Microinjection of a cell body excitant L-glutamate (44-200 nl, 0.5 M) into the caudal Ro and Rp in 15 experiments produced significant increases in pyloric minute motility index (MMI) of 4.9 +/- 1.5 (from 1.6 +/- 0.7 preinjection to 6.5 +/- 1.8 postinjection, P less than 0.05). Mean blood pressure (MBP) decreased significantly in these animals by 12 +/- 7 mmHg (from 100 +/- 6 to 88 +/- 8 mmHg, P less than 0.05). Saline microinjection in the same sites in seven cases resulted in no significant change in pyloric MMI (-1.0 +/- 0.8) or MBP (-4 +/- 11 mmHg). In five of these experiments, a second microinjection of L-glutamate (132-240 nl) was performed into the caudal Ro and Rp after spinal cord transection. This resulted in a significant increase in pyloric MMI of 3.3 +/- 0.9 (from 1.0 +/- 0.5 preinjection to 4.3 +/- 1.1 postinjection, P less than 0.05) but no change in MBP (+1 +/- 1 mmHg). Bilateral vagotomy resulted in the abrupt cessation of the pyloric response to caudal Ro and Rp stimulation. Microinjection of L-glutamate into the rostral Rp and caudal Rm in nine experiments resulted in no significant changes in pyloric MMI (-0.4 +/- 0.8) or MBP (-10 +/- 11 mmHg). These data indicate that a population of neurons in the caudal raphe nuclei, which may contain TRH, project to the DMV. In addition, excitation of these neurons causes an increase in gastric motility that is not caused by inhibition of sympathetic outflow to the gut but rather by excitation of vagal neurons in the DMV.
Glycylsarcosine transport by epithelial cells isolated from chicken proximal cecum and rectum
Authors: Calonge M. L. , Ilundain A. , Bolufer J.
Published: December 2017

The transport of glycylsarcosine (Gly-Sar) has been investigated in cells isolated from the proximal cecum and rectum of the chick, and their characteristics have been compared with those of the jejunal dipeptide transport system. Results show that 1) cecal and rectal cells are capable of actively accumulating Gly-Sar. 2) Gly-Sar initial uptake and steady-state accumulation in cecal and rectal cells were decreased in Na(+)-free mannitol-substituted medium. However, in both cell types, as in jejunal cells, intracellular accumulations higher than one can be observed in the absence of Na+. 3) By lowering the medium pH from 7.4 to 6.0, the Gly-Sar accumulation was stimulated in both cecal and rectal cells. 4) Addition of 1 mM amiloride inhibited Gly-Sar uptake by the cecal and rectal cells. We conclude that proximal cecum and rectum have a dipeptide transport system with properties similar to those found in the jejunum.
Coupling of H(+)-K(+)-ATPase activity and glucose oxidation in gastric glands
Authors: Fryklund J. , Gedda K. , Scott D. , Sachs G. , Wallmark B.
Published: December 2017

The production of 14CO2 from uniformly labeled glucose was shown to account for the entire increase in histamine-stimulated O2 consumption in rabbit gastric glands when no other substrate was added to the medium. The increased production of CO2 was correlated to the increase in O2 consumption and the accumulation of [14C]-aminopyrine (AP) after stimulation with several secretagogues. Inhibitors of H(+)-K(+)-ATPase reduced the secretagogue-induced increase in CO2 production by greater than 90%, showing that the activity of this enzyme was responsible for the greater part of gastric gland metabolism under stimulated conditions. In contrast to AP accumulation, inhibition of CO2 production by omeprazole, an acid-activated inhibitor of the H(+)-K(+)-ATPase, was not reversed by washing. The reversal of AP accumulation after omeprazole treatment and washing was most likely due to a recruitment of residual pumps bordering a nonacidic space, which had not previously been inhibited by omeprazole. These residual pumps slowly generate a pH gradient and hence AP uptake. Adding NH4+ to gastric glands resulted in a concentration-dependent increase of CO2 production up to the maximal stimulated level but without formation of the pH gradient as measured by AP uptake and loss of the omeprazole inhibition of glucose oxidation. As NH4+ can act as a K+ surrogate for H(+)-K(+)-ATPase, and as NH3 is membrane permeant, full stimulation of CO2 production is evidence that the major mechanism of H(+)-K(+)-ATPase activation in situ is an increase in the KCl permeability of the pump membrane.
Chemical degeneration of intestinal nerves
Authors: Frantzides C. T. , Garancis J. C. , Doumas B. T. , Condon R. E.
Published: December 2017

In 15 dogs, cobalt chloride solutions were infused close intra-arterially to perfuse a short segment of the jejunum. In an additional four dogs, the jejunum was perfused with the aqueous vehicle (perfusion control). All animals were killed after 1 mo and tissue samples from cobalt-treated and from nonperfused intestine (tissue comparison control) were obtained for electron microscopic and immunohistochemical studies. Segments infused with 0.25 g/dl cobalt solution showed minimal changes; the most striking feature was an increase of vasoactive intestinal polypeptide (VIP)- and substance P-containing neurosecretory granules. Cobalt chloride at higher concentrations (0.75-1.5 g/dl) induced degeneration of ganglion cells and axons in both the myenteric and submucosal plexi. In contrast, the smooth muscle and the mucosal cells of the cobalt-perfused intestine showed no histological abnormalities. Immunohistochemical staining of tissues treated with 0.75-1.5 g/dl cobalt solutions revealed absence of substance P, Met-enkephalin, and VIP immunoreactivity in all section studied; control segments showed the presence of all three peptides. Cobalt chloride in concentrations of 0.75-1.5 g/dl causes degeneration of intestinal intramural nerves and provides an experimental model suitable for studying the role of these nerves in small intestinal function.
An in vitro model of ischemia/reperfusion-induced microvascular injury
Authors: Inauen W. , Granger D. N. , Meininger C. J. , Schelling M. E. , Granger H. J. , Kvietys P. R.
Published: December 2017

The major objective of this study was to develop an in vitro model of ischemia/reperfusion (I/R)-induced microvascular injury. Cultured venular endothelial cells were grown to confluency, labeled with 51Cr, and exposed to different durations of anoxia (0.5, 1, 2, 3, and 4 h). 51Cr release and cell detachment (indexes of cell injury) were determined at different times after reoxygenation (1, 2, 4, 6, 8, and 18 h). Because in vivo studies have implicated neutrophils in I/R injury, in some experiments human neutrophils were added to the endothelial cells upon reoxygenation. Periods of anoxia greater than or equal to 2 h resulted in 70-80% 51Cr release and 80-95% cell detachment upon reoxygenation. Under these conditions (near maximal injury), the addition of neutrophils produced negligible effects. Periods of anoxia less than or equal to 1 h resulted in 30-40% 51Cr release and 50-60% cell detachment. Under these conditions (moderate cell injury), addition of neutrophils enhanced endothelial cell injury. Using a 30-min period of anoxia, we also assessed the effects of superoxide dismutase (SOD; 300 U/ml) and allopurinol (20 microM) on anoxia/reoxygenation (A/R)-induced injury in the presence or absence of neutrophils. In the absence of neutrophils, SOD or allopurinol did not protect against A/R-induced injury. However, in the presence of neutrophils, both SOD and allopurinol attenuated the increases in 51Cr release. The results derived using this in vitro model of I/R injury are largely consistent with published in vivo studies. Thus this in vitro model may provide further insights regarding the mechanisms involved in I/R injury.
Bile acids do not mediate the hyperdynamic circulation in portal hypertensive rats
Authors: Genecin P. , Polio J. , Colombato L. A. , Ferraioli G. , Reuben A. , Groszmann R. J.
Published: December 2017

Portal hypertension is accompanied by hyperdynamic systemic and splanchnic circulation. Serum bile acids (BAs), which are elevated in portal hypertension and have vasodilatory properties, have been proposed as mediators of this hyperdynamic circulation. In this study, portal hypertensive rats [accomplished by partial portal vein ligation (PVL)] were gavaged with cholestyramine (PVL-CH) to decrease circulating BA levels. A control group of rats was gavaged with an inert suspension of Metamucil (PVL-ME). The following hyperdynamic parameters were found to be similar in PVL-CH and PVL-ME: mean arterial pressure (119 +/- 6 vs. 124 +/- 5 mmHg), portal pressure (13.2 +/- 0.6 vs. 14.5 +/- 0.5 mmHg), cardiac index (0.33 +/- 0.04 vs. 0.34 +/- 0.03 ml.min-1.g body wt-1), splanchnic blood flow (1.4 +/- 0.13 vs. 1.6 +/- 0.1 ml.min-1.g body wt-1), portosystemic shunting (82 +/- 8 vs. 92 +/- 3%), peripheral arteriolar resistances (344 +/- 74 vs. 387 +/- 29 mmHg.min.ml-1.g body wt), and splanchnic arteriolar resistances (75 +/- 14 vs. 72 +/- 6 mmHg.min.ml-1.g splanchnic wt; 1,471 +/- 150 vs. 1,325 +/- 120 mmHg.min.ml-1.g body wt). BA in PVL-ME (84 +/- 9 microM/l) were similar to those previously observed in untreated PVL and significantly greater than those measured in PVL-CH (25 +/- 4 microM/l; P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)
NaCl transport across equine proximal colon and the effect of endogenous prostanoids
Authors: Clarke L. L. , Argenzio R. A.
Published: December 2017

In contrast to in vivo findings, the equine proximal colon fails to demonstrate significant net absorption of Na+ and Cl- under in vitro conditions. The present study was undertaken to determine if endogenous prostanoids are responsible for this apparent lack of ion transport. Proximal colonic tissues from ponies were preincubated in either normal Ringer solution or in Ringer containing 1 microM indomethacin and studied in Ussing chambers containing these solutions. Untreated colonic mucosa demonstrated negligible Na(+)-Cl- absorption in the basal state. In contrast, indomethacin-treated colon significantly absorbed Na+ and Cl-, primarily as the result of an equivalent increase in the mucosal-to-serosal flux of these ions. Preincubation of proximal colon in 0.1 mM ibuprofen-treated Ringer yielded similar results. Treatment of indomethacin colon with 1 mM mucosal amiloride eliminated net Na(+)-Cl- absorption without affecting the short-circuit current (Isc). The Isc in control tissue was significantly greater than in indomethacin-treated tissue and was reduced by 0.1 mM serosal furosemide. Serosal addition of 0.1 microM prostaglandin E2 or 10 mM serosal plus mucosal theophylline to indomethacin-treated tissues abolished net Na(+)-Cl- absorption and increased the Isc to levels indistinguishable from control. In contrast, control tissues were essentially unaffected by these secretagogues. These findings indicated that Na(+)-Cl- absorption in equine proximal colon was electroneutral (possibly involving Na(+)-H+ exchange) and that the tissue was capable of electrogenic Cl- secretion. However, under the in vitro conditions, basal ion transport was dominated by endogenous prostanoids that abolished Na(+)-Cl- absorption and elicited near-maximal electrogenic Cl- secretion.
Effects of ileal infusions of nutrients on motor patterns of canine small intestine
Authors: Siegle M. L. , Schmid H. R. , Ehrlein H. J.
Published: December 2017

In the present study, effects of ileal infusions of nutrients on motor patterns of the proximal small intestine and on gastric emptying were investigated in dogs. An acaloric meal was administered orally, and equicaloric loads of amino acids, oleate, and glucose were infused into the ileum at different doses (0.3, 0.6, and 0.9 kJ/min). The computerized analysis of motor patterns was focused on the differentiation between stationary and propagated contractions recorded by closely spaced extraluminal strain gauges. All three nutrients exerted inhibitory effects on gastric emptying and on contraction force and frequency of the proximal small intestine. Additionally, the propulsive motor pattern induced by the acaloric meal was modulated by reducing the number of contraction waves and their length of spread. All the effects were dose dependent. Among the three nutrients, glucose significantly changed motility at lower doses compared with amino acids and oleate. We conclude that in dogs the ileal brake mechanism is induced by all three nutrients and that it influences not only contraction force and frequency but also the motor patterns of the proximal small intestine.
In vivo transport kinetics and distribution of taurocholate by rat ileum and jejunum
Authors: Lewis M. C. , Root C.
Published: December 2017

The transport kinetics and distribution of taurocholate (TC) from the ileum and jejunum were evaluated in anesthetized Sprague-Dawley rats. Uptake and transport kinetics were determined after simultaneous administration of equimolar solutions of [3H]taurocholic acid ([3H]TC) in the ileum and [24-14C]taurocholic acid ([14C]TC) in the jejunum of anesthetized rats. At TC concentrations between 0.3 and 15 mM, total ileal absorption exceeded jejunal absorption 15- to 25-fold. The apparent Km and Vmax for ileal absorption of TC were 5.6 mM and 65.5 nmol.min-1.cm-1, respectively. Tissue distribution studies following uptake from the ileum and jejunum were done with 75Se-labeled homocholic acid taurine (75Se-HCAT). In the ileum, 82% of the 75Se-HCAT removed from the lumen was found in the bile, 10.6% was found in the ileal wall, and 7.3% was found in the liver. In the jejunum, 24.1% was found in bile, 0.6% was found in the liver, and 75.3% remained in the jejunal wall. These data show that the ileum is much more efficient and better equipped than the jejunum to take up and transport TC at concentrations up to 15 mM.
Slow waves actively propagate at submucosal surface of circular layer in canine colon
Authors: Sanders K. M. , Stevens R. , Burke E. , Ward S. W.
Published: December 2017

Colonic slow waves originate from pacemaker cells along the submucosal surface of the circular layer in the dog proximal colon. These events propagate in a nonregenerative manner into the bulk of the circular layer. Conduction velocities consistent with an active mechanism for slow-wave propagation in the longitudinal and circumferential axes of the colon have been reported. Experiments were performed using intracellular recording techniques on canine colonic muscles to determine the regenerative pathway for slow-wave propagation. In a thin band of muscle adjacent to the submucosal border of the circular layer, slow-wave amplitude was independent of distance from a pacing source, and events propagated at a rate of approximately 17 mm/s in the long axis of the circular fibers and 6 mm/s in the transverse axis of the circular fibers. These findings suggest that slow waves propagate in a regenerative manner in this region. Slow waves decayed as they conducted through regions from which the pacemaker cells had been removed with space constants of a few millimeters. Thus the integrity of the thin pacemaker region along submucosal surface is critical for propagation of slow waves and the organization of motility into segmental contractions.
Role of platelet-activating factor in ischemia/reperfusion-induced leukocyte adherence
Authors: Kubes P. , Ibbotson G. , Russell J. , Wallace J. L. , Granger D. N.
Published: December 2017

The objective of this study was to determine whether platelet-activating factor (PAF) mediates the leukocyte-endothelial cell interactions elicited by ischemia/reperfusion. The rates of adherence and extravasation of leukocytes were monitored in cat mesenteric venules subjected to 60 min of ischemia (blood flow reduced to 20% of control) followed by 60 min of reperfusion. Leukocyte rolling velocity, red blood cell velocity, and vessel diameter were also measured. The experiments were performed in control (untreated) animals and in animals pretreated with one of two PAF receptor antagonists, i.e., BN 52021 or WEB 2086. The responses of venular blood flow, wall shear rate, and vessel diameter did not differ between the three groups. In the control group, 1 h of ischemia was associated with significant adherence and extravasation of leukocytes, with reperfusion greatly enhancing these responses. The rates of leukocyte adherence and extravasation during reperfusion were greatly attenuated by both PAF antagonists. Furthermore, the proportion of adherent leukocytes that ultimately extravasate during reperfusion was markedly reduced by WEB 2086. These results suggest that PAF plays an important role in mediating the adhesive interaction between circulating leukocytes and microvascular endothelium induced by ischemia/reperfusion and that the phospholipid promotes the leukocyte extravasation associated with ischemia/reperfusion.
Changes in tone of canine gallbladder measured by an electronic pneumatic barostat
Authors: Tanaka M. , Sarr M. G. , Miller L. J. , Malagelada J. R.
Published: December 2017

We developed an electronic pneumatic barostat to measure changes in gallbladder muscle tone in vivo independent of biliary flow dynamics. Dogs were prepared with a gallbladder cannula for insertion of barostat balloon and an inflatable balloon-occluder around the cystic duct. The barostat was validated to maintain a constant pressure of 2 +/- 0.4 (SE) mmHg within a plastic balloon of “infinite” compliance that filled the gallbladder lumen; changes in balloon volume reflected changes in wall tone because intraluminal pressure was kept constant. Gallbladder tone, as measured by the barostat balloon volume, varied cyclically with the duodenal migrating motor complex and decreased by 28 +/- 8% (P less than 0.05) at the time of phase III in the stomach and duodenum. Superimposed rapid changes in barostat volume occurred during phase III. The barostat response to motilin mimicked events during spontaneous phase III; in contrast, the response to cholecystokinin octapeptide (CCK-OP) was greater (volume decrease = 53 +/- 12%) and no phasic changes occurred. Two distinct patterns of gallbladder contraction were demonstrated: a basal tone increase with superimposed phasic contractions observed during phase III and after motilin, and a basal tone increase without phasic activity that occurs after CCK-OP.
Thromboxane plays a role in postprandial jejunal oxygen uptake and capillary exchange
Authors: Alemayehu A. , Chou C. C.
Published: December 2017

The effects of a thromboxane A2 (TxA2)-endoperoxide receptor antagonist, SQ 29548, on jejunal blood flow, oxygen uptake, and capillary filtration coefficient (Kfc) were determined in anesthetized dogs under resting conditions and during the presence of predigested food in the jejunal lumen in three series of experiments. In series 1, 2.0 micrograms intra-arterial administration of SQ 29548 was found to abolish completely the vasoconstrictor action of graded doses (0.05-2.0 micrograms) of intra-arterial injection of a TxA2-endoperoxide analogue, U44069. SQ 29548 (2.0 micrograms ia) per se did not significantly alter resting jejunal blood flow, oxygen uptake, capillary pressure, or Kfc. Before SQ 29548, placement of food plus bile into the jejunal lumen increased blood flow +42 +/- 9%, oxygen uptake +28 +/- 7%, and Kfc +24 +/- 6%. After SQ 29548, the food placement increased blood flow +37 +/- 8%, oxygen uptake +52 +/- 11%, and Kfc +63 +/- 20%. The food-induced increases in oxygen uptake and Kfc after SQ 29548 were significantly greater than those induced before the blocking of TxA2-endoperoxide receptors by SQ 29548. Our study indicates that endogenous thromboxane does not play a role in regulating jejunal blood flow, capillary filtration, and oxygen uptake under resting conditions. However, it plays a role in limiting the food-induced increases in jejunal oxygen uptake and capillary exchange capacity without influencing the food-induced hyperemia.
Effect of age on the secretory capacity of pig small intestine in vivo and in vitro
Authors: McEwan G. T. , Schousboe B. , Nielsen C. G. , Skadhauge E.
Published: December 2017

The effect of age on the secretory response of pig small intestine to in vivo challenge by cholera toxin (CT) was investigated. The small intestine of 14-day-old pigs was more sensitive to CT challenge than that of 14-wk-old animals. In the 14-day jejunum CT-induced fluid secretion was five times that observed in the 14-wk tissue. Similarly, the 14-day ileum produced a fourfold higher secretion than the 14-wk ileum, although the magnitude of ileal secretion was markedly lower than that observed in the jejunum at the same CT dose. This reduced response to CT with age was not due to a reduced secretory capacity of the tissue, since supramaximal doses of prostaglandin E2 and theophylline induced a similar response in tissue from both age groups in vitro. We conclude that these results are consistent with the hypothesis that an antisecretory factor, which naturally inhibits fluid losses in enterotoxigenic diarrhea, is produced in older animals.
Na restriction blunts expansion of plasma volume and ameliorates hyperdynamic circulation in portal hypertension
Authors: Genecin P. , Polio J. , Groszmann R. J.
Published: December 2017

Expansion of plasma volume may be necessary for the development of the hyperdynamic circulation in portal hypertension. In experiment 1, sham portal vein-constricted (sham PVL) rats were divided into normal diet (NL diet sham) and low-sodium diet (low-Na sham) groups. Data obtained from the NL diet sham group was used as control data in studying portal hypertensive rats, which were also divided into normal diet (NL diet PVL) and sodium-restricted (low-Na PVL) groups on the day of portal vein constriction. There were no hemodynamic differences at 3 wk between sham PVL rats on the two diets. In contrast, sodium restriction in PVL rats resulted in a significant amelioration of the hyperdynamic circulation. In experiment 2, PVL rats were fed a normal diet for 10 days, followed by allocation to NL diet PVL and low-Na PVL groups. The hemodynamic effects of sodium restriction in these animals (studied at 22 and 28 days after PVL) were similar to those observed in experiment 1. A time course for amelioration of the hyperdynamic circulation in PVL rats by sodium restriction is described. Sodium restriction prevents the expansion of the plasma volume in PVL rats. Interference with plasma volume expansion blunts the development of (and can ameliorate) the hyperdynamic syndrome. Plasma volume expansion seems necessary for the development of the hyperdynamic circulation in portal hypertension.
Submucosal plexus alone integrates motor activity and epithelial transport in rat jejunum
Authors: See N. A. , Greenwood B. , Bass P.
Published: December 2017

It has been well established in several mammalian species, including humans, that contractions of jejunal smooth muscle correlate temporally with increases in mucosal ion transport. Furthermore, this correlation is abolished through local application of neurotoxins, suggesting interaction of enteric neurons. The purpose of this study was to determine whether the myenteric plexus is involved in this correlation. In the rat jejunum in vivo, we simultaneously measured phasic changes in intraluminal pressure and transmural potential difference (PD) as indicators of smooth muscle motor activity and epithelial ion transport, respectively. We compared the temporal association of these parameters in control animals with animals in which either the extrinsic nerves only or the extrinsic nerves and the myenteric plexus of a 5-cm jejunal segment had been ablated 30 days previously. A one-to-one coupling between muscle contractions and transmural PD fluctuations was observed in all animals; ablation of the extrinsic and/or myenteric neurons did not eliminate this correlation. We conclude that, in the rat jejunum, the submucosal plexus alone can integrate the reflex that couples ion secretion to muscle contraction.
Hypoxia-induced vasodilation of the feline superior mesenteric artery is not adenosine mediated
Authors: Lockhart L. K. , Lautt W. W.
Published: December 2017

The role of adenosine in hypoxia-induced vasodilation was examined in the intestine of pentobarbital sodium-anesthetized cats. A hollow-fiber fetal oxygenator was used to selectively reduce the PO2 of the blood supplying the superior mesenteric artery, thereby inducing hypoxia in the intestines. Decreasing the PO2 from 109 to 38 Torr caused vascular resistance to decrease from 10.2 to 7.5 Torr.kg.min.ml-1, a decrease of 2.7 Torr.kg.min.ml-1 or 24%. During selective adenosine receptor blockade with 8-phenyltheophylline, the same decrease in PO2 (from 109 to 40 Torr) produced a similar decrease in resistance from 5.7 to 3.4 Torr.kg.min.ml-1 or a difference of 2.3 Torr.kg.min.ml-1 (-36%). Thus adenosine is not the mediator of hypoxia-induced vasodilation in the feline intestine because blockade of the vasodilating effects of exogenous and presumably endogenous adenosine did not affect the observed decrease in resistance.
Rabbit ileal brush-border membrane Cl-HCO3 exchanger is activated by an internal pH-sensitive modifier site
Authors: Mugharbil A. , Knickelbein R. G. , Aronson P. S. , Dobbins J. W.
Published: December 2017

The purpose of these studies was to look for evidence of a pH-sensitive modifier site on the Cl-HCO3 exchanger on the brush-border membrane in rabbit ileum utilizing membrane vesicles. When internal pH and HCO3 were increased, Cl uptake was stimulated in a sigmoidal fashion consistent with a modifier effect. Increasing internal HCO3 alone did not have a similar effect, and increasing pH alone in the absence of HCO3 resulted in very little uptake of Cl. These results suggested that OH was a poor substrate for the exchanger but that it "activated" the transport of HCO3. Further evidence for this hypothesis was provided by the observation that increasing internal pH also stimulated Cl-Cl exchange. Altering the membrane potential with K and valinomycin had no effect on Cl uptake at high internal pH, suggesting no change in the 1:1 coupling ratio for Cl-HCO3 exchange at high internal pH. These studies provide evidence that there is an internal pH-sensitive modifier site on the Cl-HCO3 exchanger.
Characterization of saturable binding sites for circulating pancreatic polypeptide in rat brain
Authors: Whitcomb D. C. , Taylor I. L. , Vigna S. R.
Published: December 2017

Pancreatic polypeptide (PP) inhibits pancreatic exocrine secretion by indirect mechanisms that may be centrally mediated. The central site of action of PP that results in inhibition of pancreatic secretion has not been identified. Using autoradiography to identify 125I-PP binding to frozen sections of rat brain, we have identified saturable, high-affinity PP receptors in high concentrations in the interpenduncular nucleus, area postrema (AP), nucleus tractus solitarius, and dorsal motor nucleus of the vagus. The PP receptor differs from neuropeptide Y and peptide YY receptors in its binding specificity and location. Because PP is not produced in the brain, and the blood-brain barrier (BBB) excludes circulating peptides from most areas in the brain, we employed an in vivo radioreceptor assay to determine whether circulating PP binds to areas such as the AP that has both an incomplete BBB and a high concentration of PP receptors. 125I-PP and 131I-bovine serum albumin were infused simultaneously into rats through a peripheral vein with or without excess unlabeled PP. After 10 min, rats were killed and the brains were removed and cut into eight regions based on the autoradiographic localization of PP receptors. There was a significant (P less than 0.02) increase in saturable radiolabeled PP accumulation in the region that included the AP, demonstrating that circulating PP can bind to this area of the brain in vivo. PP is released into the circulation after a meal via mechanisms that exhibit vagal and cholinergic dependence. We speculate that PP completes a feedback loop by binding to receptors in the AP and interacting with the adjacent vagal nuclei to inhibit vagal activity.
Gastrin-releasing peptide directly releases pepsinogen from guinea pig chief cells
Authors: Fiorucci S. , McArthur K. E.
Published: December 2017

Gastrin-releasing peptide (GRP) and bombesin can stimulate pepsinogen release by both gastrin-dependent and -independent mechanisms. Using isolated guinea pig gastric chief cells, we determined that GRP can act directly on the guinea pig chief cell to cause pepsinogen release. GRP and bombesin stimulated a 2.5- to 3-fold increase in pepsinogen release above basal release. Substance P also stimulated a small but significant increase in pepsinogen release. No gastrin immunoreactivity was detected in the supernatants of cells stimulated with up to 1 microM GRP or bombesin or 1 mM carbachol. GRP-stimulated pepsinogen release was completely inhibited by GRP/bombesin receptor agonists as well as substance P receptor antagonist but not by antagonists to receptors for gastrin, the octapeptide of cholecystokinin (CCK-8), secretin, vasoactive intestinal peptide (VIP), or muscarinic agents. Substance P-stimulated pepsinogen release was completely inhibited by substance P receptor antagonist but not by GRP/bombesin receptor antagonists. An additive effect on pepsinogen release was seen when GRP was combined with maximally effective concentrations of adenosine 3',5'-cyclic monophosphate (cAMP)-mediated agents (VIP, secretin, 8-BrcAMP) but not with calcium-mediated agents (carbachol, CCK-8, gastrin). These results indicate that GRP can directly stimulate pepsinogen release from guinea pig chief cells by a specific GRP receptor that mobilizes intracellular calcium.
Glycyl-L-sarcosine transport by ATP-depleted isolated enterocytes from chicks
Authors: Calonge M. L. , Ilundain A. , Bolufer J.
Published: December 2017

Normally energized and ATP-depleted isolated chicken enterocytes have been used to investigate the energetics of intestinal glycyl-L-sarcosine (Gly-Sar) transport, and the results were compared with those obtained for 3-O-methyl-D-glucose (3-OMG) active transport, which is known to be energized by the electrochemical Na+ gradient. The results show that even though Gly-Sar had no effect on Na+ efflux from labeled (22Na) enterocytes, 20 mM L-leucine and 20 mM 3-OMG increased the rate constant of Na+ efflux by 34 and 76%, respectively. In ATP-depleted cells 3-OMG was accumulated in response to experimentally imposed inward Na+ gradients; however, Gly-Sar was not accumulated. In the absence of Na+, an inward proton gradient slightly increased Gly-Sar uptake, although no transient accumulation was observed. An increase in membrane potential by replacement of Cl- by NO3- or by imposition of an outward K+ gradient in the presence of valinomycin stimulated both 3-OMG and Gly-Sar uptake, even in the absence of either H+ or Na+ gradient. When in addition to a K+ diffusion potential, a Na+ gradient was present, 3-OMG transient accumulation was further increased (approximately 2-fold over equilibrium value), and Gly-Sar uptake showed the same pattern as that observed when only membrane potential was present. However, in the presence of an outward K+ diffusion potential and an inward H+ gradient Gly-Sar was stimulated approximately threefold, clearly showing a significant overshoot, whereas 3-OMG was not transiently accumulated.
Na+ transport properties of isolated rat parotid acini
Authors: Dissing S. , Nauntofte B.
Published: December 2017

The effect of carbachol stimulation on the Na+ transport properties of rat parotid acini was characterized. Upon stimulation, the acinar Na+ concentration increased from 20 to 70 mM within 25 s, whereafter a backregulation toward the prestimulatory level was observed, mediated by the Na(+)-K+ pump. The transport mechanisms responsible for the net Na+ uptake observed between 10 and 20 s after carbachol stimulation in a Krebs-Ringer-bicarbonate medium (KRB) consisted of a dimethylamiloride-sensitive Na(+)-H+ exchange system (3.5 mmol.liter cell water-1.s-1 or approximately 75% of the total Na+ influx) and a bumetanide-sensitive cotransport system (of approximately 15%). The data are consistent with the residual influx being mediated by amiloride-sensitive Na+ channels. In unstimulated acini acidified by a NH4+ prepulse technique, the relationship between intracellular pH (pHi) and the rate of acinar Na(+)-H+ exchange was determined. At pHi 6.5 the rate of Na(+)-H+ exchange in a KRB medium amounted to 1.2 mmol.liter cell water-1.s-1 and ceased when pHi had recovered to 7.2. It was concluded that under physiological conditions carbachol stimulation activates Na(+)-H+ exchange, an effect that is responsible for the major part of the Na+ gain after stimulation. The role of cotransport in mediating net Cl- uptake against an electrochemical gradient was investigated. It was found that when the Na+ gradient is strongly reduced (in a 20 mM Na+ medium) a Cl(-)-HCO3- exchange system can mediate a net Cl- uptake at a rate similar to the rate observed in a medium containing high Na+. The results are compatible with both Cl(-)-HCO3- exchange and cotransport keeping an intracellular Cl- concentration above the electrochemical equilibrium under physiological conditions.
Control of lower esophageal sphincter pressure by two sites in dorsal motor nucleus of the vagus
Authors: Rossiter C. D. , Norman W. P. , Jain M. , Hornby P. J. , Benjamin S. , Gillis R. A.
Published: December 2017

Our purpose was to determine the central vagal sites for regulating changes in lower esophageal sphincter (LES) pressure in the cat. Injection of the retrograde tracer, horseradish peroxidase, into the LES resulted in labeling of cells in the dorsal motor nucleus of the vagus (DMV), with the largest number of cells appearing in two areas, one area rostral to obex (1.5-4.0 mm) and one area caudal to obex (-0.5 to -1.5 mm). In alpha-chloralose-anesthetized cats, L-glutamic acid was microinjected into these areas and LES pressure, intragastric pressure, and stomach motility were monitored. Microinjection of L-glutamic acid into the rostral area resulted in significant increases in LES pressure (18.6 +/- 4.9 mmHg; P less than 0.05), pyloric motility (baseline minute motility increased from 5.7 +/- 2.2 to 14.5 +/- 3.9 postinjection; P less than 0.05) and stomach pressure (baseline of 16.9 +/- 2.3 mmHg increased to 23.8 +/- 3.7 mmHg postinjection; P less than 0.05). Microinjection of L-glutamic acid into the caudal area resulted in significant decreases in LES pressure (-14.3 +/- 5.8 mmHg; P less than 0.05) and intragastric pressure (-7.5 +/- 2.2 mmHg; P less than 0.05) with no significant changes in pyloric motility. Ipsilateral vagotomy abolished both sets of responses. These data indicate that excitatory and inhibitory control of LES and intragastric pressure are mediated by vagal efferent neurons located in two distinct sites in the DMV.
Superoxide anion generation by in situ perfused rat liver: effect of in vivo endotoxin
Authors: Bautista A. P. , Spitzer J. J.
Published: December 2017

A simple method is described to monitor the superoxide dismutase (SOD)-inhibitable production of superoxide anion (O2-.) in the liver. The isolated rat liver was perfused in situ with ferricytochrome c, and the reduction of this substrate during perfusion was determined. Within 30 s after the introduction of the substrate, significant reduction of ferricytochrome c was observed and stabilized at 2-4 min. A marked reduction of the substrate was observed in the livers of rats that received Escherichia coli lipopolysaccharide (LPS, 1 mg/kg) in vivo 3 h before liver perfusion. Ferricytochrome c reduction was inhibited by SOD, but not significantly with allopurinol or deferoxamine mesylate in the livers of LPS-treated rats. Control livers exhibited only a small reduction of the substrate, and this was not significantly inhibited by SOD. After in vivo LPS administration, O2-. production peaked in the liver at 3 h (6.6 nmol/min) and returned toward normal at 6 h (1 nmol/min) after endotoxin. The amount of O2-. generated by the endotoxic livers was dose related. At 3 h post-LPS, neutrophil infiltration and necrotic areas were found in the histological sections of the liver with concomitant elevation of serum aminotransferases, indicating hepatic abnormalities during the early stage of endotoxemia. Phorbol myristate acetate in the perfusion system markedly enhanced O2-. generation in the endotoxic liver. These results show that the perfused rat liver can be used to measure O2-. generation following in vivo stimuli. The data also demonstrate that O2-. release after LPS treatment in vivo is a short and early event and may have an important role in hepatic injury in endotoxemic conditions.
Taurocholate induces pericanalicular localization of C6-NBD-ceramide in isolated hepatocyte couplets
Authors: Crawford J. M. , Vinter D. W. , Gollan J. L.
Published: December 2017

The mechanisms and pathways involved in hepatocellular transport of lipid destined for biliary excretion remain poorly understood. Using fluorescence microscopy of rat hepatocyte couplets in primary culture, we examined the effects of taurocholate (TC) on the intracellular distribution of 6-N-[7-nitrobenz-2-oxa-1,3-diazol- 4-yl]aminocaproyl-sphingosine (C6-NBD-ceramide), a lipid that accumulates in the Golgi apparatus. Microscopic findings were quantified with morphometric and digital image analysis and were correlated with the metabolism of C6-NBD-ceramide in isolated hepatocyte suspensions and the biliary excretion of fluorescent lipid in the isolated perfused liver. After plasma membrane uptake of C6-NBD-ceramide from albumin at 0 degrees C, the lipid was rapidly internalized at 37 degrees C but exhibited only a modest concentration of fluorescence in intracellular organelles. With 17 microM TC in the medium, there was enhanced localization of fluorescence to organelles and significant recruitment of fluorescent lipid to the pericanalicular region of the couples within 30 min. C6-NBD-ceramide was partially metabolized to C6-NBD-sphingomyelin and -glucosylceramide, indicative of transit through the Golgi apparatus. The generation of C6-NBD-sphingomyelin was significantly increased by TC. After a similar loading protocol in the perfused liver, there was little biliary excretion of fluorescent lipid at 37 degrees C under control conditions. However, infusion of TC markedly enhanced the biliary output of fluorescent lipid over the first 30 min, primarily as C6-NBD-sphingomyelin and -glucosylceramide. We conclude that TC modulates the distribution of C6-NBD-ceramide in hepatocytes by inducing translocation of lipid to a pericanalicular location, most likely the Golgi apparatus, before excretion of its metabolites in bile. Our findings support the concept that bile salt-induced biliary lipid excretion is facilitated by the interaction of bile salts with lipids at the level of intracellular organelles.
Hemodynamic characterization of conscious and ketamine-anesthetized bile duct-ligated rats
Authors: Sikuler E. , Buchs A. E. , Yaari A. , Keynan A.
Published: December 2017

The present study was conducted to characterize the hemodynamic alterations in common bile duct-ligated (CBDL) rats under ketamine anesthesia and in the awake restrained state. Hemodynamic studies using the radioactive microspheres technique were performed 17.6 +/- 0.6 (SE) days after bile duct ligation or sham operation. CBDL rats had lower mean arterial pressure, reduced systemic and renal resistance, and increased renal blood flow compared with sham-operated rats. This was found both in the conscious and anesthetized states. Anesthetized CBDL rats had higher portal pressure (13.2 +/- 0.5 vs. 9.2 +/- 0.4 mmHg; P less than 0.001) and lower splanchnic arteriolar resistance (15.4 +/- 1.3 vs 26.8 +/- 4.6 mmHg.ml-1.min.100 g body wt; P less than 0.05) than sham-operated rats. Portosystemic shunting was 52.3 +/- 11.7% in CBDL and negligible in sham-operated rats. The last three parameters could not be measured in conscious animals. Total peripheral resistance was lower in the conscious than in the anesthetized state, diverting a higher fraction of cardiac output at the expense of splanchnic organs and leading to a significant reduction of portal venous inflow in sham-operated but not in CBDL rats [3.36 +/- 0.47 vs. 5.38 +/- 0.65 (P less than 0.05) and 5.33 +/- 0.58 vs. 6.34 +/- 0.37 ml.min-1.100 g body wt-1 (P = NS), respectively]. These findings indicate that CBDL and normal rats respond differently to anesthesia and restraint. Because the restrained state is stressful and studies in anesthetized animals are technically simpler, provide additional information such as portal pressure and portosystemic shunting, and diminish animal suffering, we suggest that hemodynamic studies in rats, using the microsphere technique, should be preferably performed under ketamine anesthesia.