bluuebunny/crossref_metadata_2025 huggingface dataset:
Compensatory mechanisms in response to an elevated hepatic oxygen consumption in chronically ethanol-fed rats
Authors: Bredfeldt J. E. , Riley E. M. , Groszmann R. J.
Published: December 2017

During conditions that elevate hepatic oxygen consumption (VLO2), oxygen delivery, or oxygen extraction may also increase, acting as compensatory mechanisms. VLO2 was quantitated by an in vivo method in chronically ethanol-fed rats to establish whether VLO2 was increased and what compensatory mechanisms might ensue. VLO2 was increased 45% (0.32 +/- 0.02 ml O2 X min-1 X 100 g body wt-1; P less than 0.001) in rats chronically fed ethanol for 8 wk, while VLO2 (0.28 +/- 0.04; P = NS) was not increased in chronically ethanol-fed rats withdrawn from ethanol 20 h before study compared with control rats (0.22 +/- 0.01). Oxygen delivery was increased 31% (P less than 0.05) in ethanol-fed rats and adequately compensated for the increased VLO2. Hepatic artery blood flow did not increase in ethanol-fed rats, indicating a lack of hepatic artery vasodilation in response to the elevated VLO2. As a result, a greater percentage of oxygen delivery was supplied via portal venous blood flow that has a reduced oxygen content. These observations might suggest that ethanol-fed rats may have a decreased reserve for maintaining an adequate oxygen supply to the liver and may be at an increased liability for developing hepatic hypoxia if oxygen delivery and/or extraction were compromised.
Characteristics of sudden potential drop in bullfrog gastric mucosa
Authors: Matthews J. B. , Rangachari P. K. , Rowe P. H. , Lange R. , Marrone G. , Silen W.
Published: December 2017

An unusual sudden potential drop (SPD) has been described by Kidder in bullfrog gastric mucosa exposed to anoxia and a serosal pH less than or equal to 7.1. We found that anoxia was not a prerequisite, since under fully oxygenated conditions the SPD occurred reliably in metiamide-treated tissues when the pH of the nutrient solution (pHN) was below 7.1. The SPD was observed also in metiamide-treated tissues exposed to 20 mM luminal or nutrient acetylsalicylic acid (ASA) with pHN = 7.3, an effect that was abolished by increased nutrient HCO3- concentration. The SPD occurred when NO3- but not isethionate or acetate replaced Cl- in the bathing media. A marked increase in potential difference in response to changing luminal Cl- concentration was observed after the SPD, uptake of Cl- from the luminal solution into the tissue increased, but transmural fluxes of Cl- decreased bidirectionally. Permeability to H+ was unaltered in the post-SPD state. An SPD never occurred in antrum under conditions causing SPD in fundus, suggesting that oxyntic cells are prerequisite. We conclude that, under conditions which cause tissue or cellular acidosis and cessation of H+ secretion, fundic mucosae respond with an anion-selective increase in apical permeability manifest as the SPD.
Mechanisms maintaining a low-pH microclimate in the intestine
Authors: Shiau Y. F. , Fernandez P. , Jackson M. J. , McMonagle S.
Published: December 2017

The presence of an unequilibrated region adjacent to the mucosal surface of the small intestine has long been suggested. Controversy remains as to the characteristics of this region. This study was designed to further define these characteristics. Our studies demonstrated by two independent methods, pH electrode measurement and dye indicator, that a low-pH compartment does exist at the surface of the intestine. The thickness of this microclimate was estimated to be 700 microns, comparable with that described for the unstirred water layer. Maintenance of the low-pH compartment is due to the presence of a mucus coating rather than hydrogen ion secretion. We postulate that the mucus functions as an ampholyte and restricts hydrogen ion movement in its matrix. The acidic microclimate, unstirred water layer, and the mucus coating might in fact represent a common phenomenon.
Taurocholate transport by basolateral plasma membrane vesicles isolated from developing rat liver
Authors: Suchy F. J. , Courchene S. M. , Blitzer B. L.
Published: December 2017

Taurocholate transport was characterized in basolateral plasma membrane vesicles prepared from the livers of 14-day-old Sprague-Dawley rats using a self-generating Percoll gradient method. Liver plasma membrane protein yield, intravesicular volume, and enrichments of various marker enzymes were similar to those obtained for vesicles from adult rat liver. The basolateral marker enzyme Na+-K+-ATPase was enriched 26-fold in the suckling rat basolateral membrane fraction while the bile canalicular marker enzymes alkaline phosphatase and Mg2+-ATPase were enriched only 3- and 5-fold, respectively. The activities of marker enzymes for endoplasmic reticulum, mitochondria, or lysosomes were not enriched compared with homogenate. In the presence of an inwardly directed 100 mM Na+ gradient, vesicle accumulation of taurocholate transiently reached a concentration 1.5- to 2-fold higher than that at equilibrium ("overshoot") in suckling and adult membrane vesicles, but the initial rate of taurocholate entry and peak intravesicular accumulation were markedly decreased in suckling compared with adult membrane vesicles. In the presence of an inwardly directed 100 mM K+ gradient, the rate of uptake was slower, and no overshoot occurred in either suckling or adult rat vesicles. The decreased rate of Na+-coupled taurocholate uptake by membrane vesicles from suckling rat liver could not be explained on the basis of more rapid dissipation of the transmembrane Na+ gradient. Kinetic studies demonstrated saturable, Na+-dependent taurocholate uptake for both suckling and adult vesicles. However, the Vmax for taurocholate uptake in suckling rat vesicles was less than half of the adult rate (2.46 +/- 0.13 vs. 5.25 +/- 0.22 nmol X mg prot-1 X min-1, respectively, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)
Preferential binding of vasoactive intestinal peptide to hepatic nonparenchymal cells
Authors: Gardner D. F. , Kilberg M. S. , Wolfe M. M. , McGuigan J. E. , Misbin R. I.
Published: December 2017

The binding of 125I-vasoactive intestinal peptide (125I-VIP) and 125I-insulin has been examined in highly enriched populations of rat hepatocytes and hepatic nonparenchymal cells. 125I-VIP bound to high-affinity sites (Ka = 1.7 X 10(9) M-1) in nonparenchymal cells. Specific binding in these cells was nearly fivefold greater than in hepatocytes (15.0 +/- 0.6% vs. 3.6 +/- 0.7% radioactivity bound per 4 X 10(5) cells). In contrast, 125I-insulin binding was similar in both cell populations (18.2 +/- 2.4% per 4 X 10(5) cells in hepatocytes vs. 17.1 +/- 1.0% in nonparenchymal cells). Glucagon and insulin had no effect on 125I-VIP binding in nonparenchymal cells. Secretin inhibited 125I-VIP binding but was only about 1% as potent as unlabeled VIP. VIP had no apparent effect on cAMP levels in either cell population, whereas glucagon increased cAMP levels in both cell types. Our findings suggest that VIP binds preferentially to hepatic nonparenchymal cells and that these cells are primarily responsible for the clearance of VIP from the portal circulation.
Transport of glycyl-L-proline by mouse intestinal brush-border membrane vesicles
Authors: Rajendran V. M. , Berteloot A. , Ramaswamy K.
Published: December 2017

The characteristics of [14C]glycyl-L-proline transport have been studied using brush-border membrane vesicles from mouse small intestine in order to investigate the transport of nonhydrolyzable peptide across the brush-border membrane. Uptake curves for the peptide did not exhibit overshoot phenomena and were similar under Na+ or K+ gradient conditions (extravesicular greater than intravesicular). However, L-proline was transported by Na+ gradient-dependent system. Analysis of the incubation medium and the intravesicular contents showed that there was negligible hydrolysis of the peptide. Transport of glycyl-L-proline was saturable, conforming to Michaelis-Menten kinetics with a Km of 30.8 +/- 1.9 mM and a Vmax of 5.96 +/- 0.17 nmol.mg prot-1.0.4 min-1. Uptake of glycyl-L-proline was not significantly inhibited by free amino acids nor by most of the peptides containing D amino acids but was strongly inhibited (up to 64%) by various di- and tripeptides of L amino acids. These results clearly show that glycyl-L-proline was transported by a Na+-independent, carrier-mediated process. Our results suggest that the nonhydrolyzable peptides are transported mostly by carrier-mediated processes in contrast to hydrolyzable peptides.
Studies on role of calmodulin in Ca2+ regulation of rabbit ileal Na and Cl transport
Authors: Donowitz M. , Wicks J. , Madara J. L. , Sharp G. W.
Published: December 2017

To study the role of calmodulin in regulation of rabbit ileal active electrolyte transport by Ca2+, the effects of the Ca2+-calmodulin antagonist naphthalenesulfonamides W12 and W13 and the weak Ca2+-calmodulin antagonist promethazine, a phenothiazine, were studied on basal ileal Na and Cl transport and on secretion stimulated by Ca2+ and by cAMP. The naphthalenesulfonamides and promethazine all stimulated basal ileal Na and Cl absorption. The stimulation of Na absorption was dependent on Cl in the bathing solutions, and the stimulation of Cl absorption was Na dependent. This suggests that the transport process stimulated was the neutral, linked NaCl absorptive process. W13, which is a better Ca2+-calmodulin antagonist but has similar hydrophobic properties to W12, stimulated active ileal absorption at a lower concentration than W12, suggesting that the naphthalenesulfonamide-induced stimulation of active ileal absorption was not due to the hydrophobic properties of these drugs but could be due to their effects on the calcium-binding protein calmodulin. W12, W13, and promethazine did not alter paracellular transport, not affecting dilution potentials or structural features of the paracellular pathway. W12 and W13 did not decrease the changes in active ileal Na and Cl transport caused by increasing ileal cAMP content or intracellular Ca2+. This suggests that calmodulin is not directly involved in the active electrolyte secretion caused by increased intestinal Ca2+ or cAMP.
Rat gastric mucosal microcirculation in vivo
Authors: Holm-Rutili L. , Obrink K. J.
Published: December 2017

The superficial gastric mucosal microcirculation was observed microscopically by transillumination in the anesthetized rat. The vessels surrounding the gastric crypts were monitored on a television screen through a microscope and the pictures stored on a videotape for off-line analysis of red cell velocity (VRBC) and vessel diameter. From these measurements microvascular volume flows were calculated. VRBC reached steady values after 1-4 h (mean 2 h) and showed a regular pulsatile flow (4-7 cycles/min) in most experiments. Acid output was measured at regular intervals; 50% of the rats showed no spontaneous acid output, but the others secreted up to 100 mu eq/h. The microvessels in the superficial mucosa were classified into three orders according to their branching hierarchy and relative dimensions, and their distribution per unit mass was estimated. VRBC and volume flow were shown to decrease in the successive orders of the microvessels. Calculation of organ blood flow from microvascular flow data and vessel distribution gave values (21 ml.min-1.100 g tissue-1) that agree with earlier reported values. A higher flow velocity was detected in rats with spontaneous acid output than in those without, but there was a poor correlation between the magnitude of the acid output and VRBC. Pentagastrin (96 micrograms.kg-1.h-1) induced a significant increase in both blood flow and acid secretion. Results from this study indicate that this experimental model is potentially useful for studies of the correlation between acid secretion and mucosal blood flow.
Rapid and repeated blood sampling in the conscious laboratory rat: a new technique
Authors: Winsett O. E. , Townsend C. M. , Thompson J. C.
Published: December 2017

A method for obtaining repeated blood samples from conscious rats by translumbar vena cava puncture is presented. Studies performed at laparotomy enabled us to select a 5/8-in., 25-gauge needle as the ideal instrument for aspirating blood. The awake rat is held prone by an assistant and the needle inserted at the level of the first lumbar vertebra in the coronal plane at an angle of 45 degrees from the vertical. We performed the procedure 350 times in 30 rats (100–300 g) at weekly intervals for 8 wk. Each time, we have obtained 2 ml of blood. The time required for obtaining 30 samples is 15–20 min. There have been no complications and only three deaths (mortality rate, 0.9% per puncture); in all three instances the rats jerked free, lacerating the vena cava. The procedure has been performed in guinea pigs and hamsters, with equal ease, without morbidity or mortality. This new technique permits rapid, atraumatic, repeated samplings from awake animals with no morbidity and minimal mortality.
Receptor for calcitonin gene-related peptide: binding to exocrine pancreas mediates biological actions
Authors: Seifert H. , Sawchenko P. , Chesnut J. , Rivier J. , Vale W. , Pandol S. J.
Published: December 2017

In the present study we demonstrate by immunohistochemical techniques that calcitonin gene-related peptide (CGRP) is present in nerve terminals in the islets of Langerhans. Furthermore, binding studies with 125I-CGRP indicate that dispersed acini from guinea pig pancreas contain a single class of high-affinity binding sites for CGRP with an apparent dissociation constant of 18 nM. Vasoactive intestinal peptide (VIP), rat growth hormone-releasing factor (rGRF), cholecystokinin octapeptide (CCK-OP), and bombesin do not interact with these receptors. Interaction of CGRP with these receptors leads to release of amylase from the acinar cells. Amylase release is half maximal at 0.3 nM CGRP and maximal at 3 nM CGRP. Maximal amylase release with CGRP is one-third of that observed with VIP. CGRP-induced amylase release is dependent on theophylline in the incubation medium. CGRP potentiates the amylase release stimulated by bombesin and CCK-OP but has no effect on amylase release stimulated by VIP, rGRF, and natural glucagon. CGRP stimulates a 25% increase in basal cellular cAMP. These results indicate that guinea pig pancreatic acinar cells contain a novel receptor for CGRP and that interaction of CGRP with this receptor leads to digestive enzyme secretion through a cAMP-mediated pathway. The presence of CGRP in the islets of Langerhans suggests a pathway for CGRP to reach the exocrine pancreas through an insuloacinar portal system.
Duodenal acidification releases cholecystokinin
Authors: Chen Y. F. , Chey W. Y. , Chang T. M. , Lee K. Y.
Published: December 2017

We studied the effect of hydrochloric acid (HCl) in the proximal small intestine on release of cholecystokinin (CCK) and secretin and on exocrine pancreatic secretion in conscious dogs with gastric cannulas and modified Herrera pancreatic cannulas. Intraduodenal administration of HCl in a concentration of 50 or 100 mM at rates of 0.05, 0.1, 0.2, and 0.4 mmol/min significantly increased plasma concentration of CCK in a dose-dependent manner, whereas plasma gastrin levels decreased. The increased plasma CCK level paralleled a significant increase in pancreatic trypsin output. Plasma secretin concentration and pancreatic bicarbonate output also increased in response to the acid, and the increase was dependent on the acid loads delivered in the duodenum. Thus, in dogs, HCl in the duodenum releases both CCK and secretin to stimulate pancreatic secretion of bicarbonate as well as enzymes.
Effect of systemic acid-base disorders on colonic intracellular pH and ion transport
Authors: Wagner J. D. , Kurtin P. , Charney A. N.
Published: December 2017

We have previously reported that changes in colonic net Na and Cl absorption correlate with arterial CO2 partial pressure (PCO2) and that changes in colonic net Cl absorption and HCO3 secretion correlate with the plasma HCO3 concentration during the systemic acid-base disorders. To determine whether changes in intracellular pH (pHi) and HCO3 concentration [( HCO3]i) mediate these effects, we measured pHi and calculated [HCO3] in the distal colonic mucosa of anesthetized, mechanically ventilated Sprague-Dawley rats using 5,5-[14C]dimethyloxazolidine-2,4-dione and [3H]inulin. Rats were studied during normocapnia, acute respiratory acidosis and alkalosis, and uncompensated and pH-compensated acute metabolic acidosis and alkalosis. When animals in all groups were considered, there were strong correlations between mucosal pHi and both arterial PCO2 (r = -0.76) and pH (r = 0.82) and between mucosal [HCO3]i and both arterial PCO2 (r = 0.98) and HCO3 concentration (r = 0.77). When we considered the rates of colonic electrolyte transport that characterized these acid-base disorders [A. N. Charney and L. P. Haskell. Am. J. Physiol. 246 (Gastrointest. Liver Physiol. 9): G159-G165, 1984], we found strong correlations between mucosal pHi and net Na absorption (r = -0.86) and between mucosal [HCO3]i and both net Cl absorption (r = 0.98) and net HCO3 secretion (r = 0.83). These findings suggest that the systemic acid-base disorders cause changes in colonic mucosal pHi and [HCO3]i as a consequence of altered arterial PCO2 and HCO3 concentration. In addition, the effects of these disorders on colonic electrolyte transport may be mediated by changes in mucosal pHi and [HCO3]i.
Characterization of calcium deprivation-induced cholestasis in the perfused rat liver
Authors: Reichen J. , Berr F. , Le M. , Warren G. H.
Published: December 2017

To elucidate the role of calcium in regulation of canalicular bile flow, we studied biliary sucrose permeability and the transport kinetics of taurocholate in the in situ perfused rat liver. Calcium deprivation did not adversely affect viability or ultrastructural appearances of the liver. Removal of calcium led to initial choleresis followed by cholestasis dependent on external ionized calcium concentration. Biliary recovery of [14C]sucrose relative to that of tritiated water was determined by a biliary multiple-indicator dilution technique. Analysis in terms of irreversible thermodynamics suggested that biliary permeability to sucrose increased due to a change in the sieving coefficient from 0.135 to 0.435. Biliary recovery of taurocholate was significantly (P less than 0.001) reduced in low-calcium medium (from 79.6 +/- 6.5 to 17.6 +/- 11.8%). This was not due to a defect in hepatocellular uptake of taurocholate as determined in the perfused rat liver by a multiple-indicator dilution technique and in isolated hepatocytes. We conclude that calcium deprivation-induced cholestasis is characterized by an increased biliary permeability, a defect in cellular translocation and/or canalicular secretion of bile salts, and a defect in bile salt-independent bile flow.
Phosphate transport in intestinal brush-border membrane vesicles: effect of pH and dietary phosphate
Authors: Quamme G. A.
Published: December 2017

Phosphate transport was investigated in brush-border membrane vesicles prepared from the duodenal segment of the chicken small intestine. Phosphate transport was sodium dependent and demonstrated the typical overshoot phenomenon with an external sodium gradient. There was no appreciable difference in phosphate uptake rates at pH 6.5 or 7.5 values, and the apparent kinetic parameters suggested that both the monovalent form, H2PO4-, and divalent form, HPO4(2-), were transported at the appropriate pH values. The apparent Km values were 0.10 and 0.07 mM, respectively. Determination of the sodium stoichiometry, assessed by an external sodium concentration or by the static-head method, suggested an interaction of one or more sodium ions with phosphate at the acid pH value and two or more sodium ions with phosphate at the alkaline pH value. The sodium affinity for sodium-dependent phosphate transport was in the order of 10–25 mM and was similar at both pH values. Phosphate accumulation was not affected by an externally applied potassium diffusion potential, suggesting an electroneutral process at both pH values. The cumulative evidence would suggest Na+-H2PO4- transport at pH 6.5 and 2Na+-HPO4(2-) at pH 7.5 value, and the effect of hydrogen ions on sodium-dependent phosphate transport is dependent on the external sodium concentration. Brush-border membranes isolated from chickens maintained for 5–7 days on a low-phosphate diet demonstrated a greater sodium-dependent phosphate uptake (Vmax) than animals on normal phosphate intakes with no change in the apparent Km. Sodium stoichiometry and the rheogenic nature of the process appear to be similar to those present in normal birds.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of dietary cellulose on site of lipid absorption
Authors: Gallaher D. , Schneeman B. O.
Published: December 2017

The effect of dietary cellulose on the localization within the small intestine of isotopically labeled triglyceride (TG) and cholesterol (CH) from a test meal was investigated. Feeding a 20% cellulose meal resulted in greater quantities of 14C-TG present in both the contents and mucosa of the distal intestine compared with a fiber-free control meal. In contrast, cellulose had no effect on the localization of CH within either the intestinal contents or the mucosa. Accumulation of TG within the intestine was not due to differences in stomach emptying, as the emptying rate was similar for both TG and CH. Within the bulk phase TG must be hydrolyzed by pancreatic lipase before it is available for cellular uptake at the microvillus membrane, whereas CH requires no hydrolysis. The greater amount of TG, but not of CH, within the intestine suggests that cellulose can interfere with lipase activity in vivo. Consequently, cellulose can delay TG hydrolysis and increase the amount of lipid absorbed in the ileum.
Uptake and release of gamma-aminobutyric acid in guinea pig gallbladder
Authors: Saito N. , Taniyama K. , Tanaka C.
Published: December 2017

The presence of gamma-aminobutyric acid (GABA)-ergic neuron in guinea pig gallbladder was investigated by measuring GABA contents and glutamate decarboxylase (GAD) activity and by demonstrating the uptake and release of [3H]GABA. GABA and GAD are both present in the gallbladder, and a positive correlation in regional distribution was observed among GABA, GAD, and the number of ganglion cells. The uptake of [3H]GABA by the gallbladder showed two saturable components; a high-affinity component (Km = 23.3 microM, Vmax = 7.63 nmol X g-1 X 10 min-1) and a low-affinity component (Km = 515 microM, Vmax = 57.1 nmol X g-1 X 10 min-1). These high-affinity and low-affinity transport systems corresponded to those obtained in the presence of beta-alanine and L-2,4-diaminobutyric acid, respectively, thereby suggesting the presence of neuronal and nonneuronal GABA transport systems in this tissue. Electrical transmural stimulation produced an increase in [3H]-GABA release from the isolated gallbladder preloaded with [3H]GABA, in the presence of beta-alanine. The stimulation-evoked release of [3H]GABA was prevented by calcium-free medium containing 1 mM EGTA and tetrodotoxin, thereby indicating that the released GABA originates from the nerve terminals. These results provide evidence for the presence of GABA-ergic neurons in the guinea pig gallbladder.
Effect of GRP on beta-adrenergic-stimulated gastrin and somatostatin release in the isolated rat stomach
Authors: Short G. M. , Reel G. M. , Doyle J. W. , Wolfe M. M.
Published: December 2017

The present studies were directed toward examining the effects of gastrin-releasing peptide (GRP) on acid secretion and on beta-adrenergic-stimulated gastrin and somatostatin release using the isolated vascularly perfused rat stomach. Including pentagastrin in perfusion buffer increased acid output from 2.2 +/- 0.4 mueq H+/h during control perfusion to 18.8 +/- 1.8 mueq H+/h (P less than 0.01). No significant changes in acid secretion were detected when either GRP or specific antibodies to GRP were included in perfusate in the absence or presence of pentagastrin. Inclusion of 10(-9) M isoproterenol in the perfusate did not change acid output with respect to control; however, gastrin and somatostatin release into the portal venous effluent was significantly enhanced. Peak gastrin and somatostatin concentrations observed at 15 min were 753 +/- 43% (P less than 0.001) and 345 +/- 43% (P less than 0.01), respectively, of basal levels. When antibodies to GRP were included in perfusate containing isoproterenol, gastrin and somatostatin release into the portal venous effluent was significantly inhibited. The results of these studies indicate that GRP does not affect basal or pentagastrin-stimulated gastric acid secretion in the isolated perfused rat stomach. However, under the conditions of these experiments, beta-adrenergic stimulation of gastrin and somatostatin release appears to be mediated, at least in part, through GRP.
Effect of lactation on cholesterol synthesis in rats
Authors: Feingold K. R. , Moser A. H.
Published: December 2017

Lactation induces a variety of morphological and functional changes in the gastrointestinal tract. In the present study we employed tritiated water as the substrate to demonstrate that in the intact rat lactation results in a twofold increase in cholesterol synthesis in the small intestine. Feeding a high-cholesterol diet did not markedly inhibit small intestinal cholesterol synthesis in either control or lactating animals, and the difference in cholesterol synthesis between the two groups persisted. In the large intestine, cholesterol synthesis is increased threefold in the lactating animals, and feeding a high-cholesterol diet did not affect synthesis in either the control or lactating animals. In the liver, lactation stimulated cholesterol synthesis, and quantitatively this increase in hepatic cholesterol synthesis is much greater than the increase observed in the intestines. Feeding the rats a high-cholesterol diet markedly inhibited hepatic cholesterol synthesis in both control and lactating animals, a finding that demonstrates that the feedback inhibition of cholesterol synthesis in the liver is not impaired by lactation. In the lactating animals, the quantity of labeled cholesterol in 1 ml of serum is 2.4 times greater than observed in controls. Feeding the rats a high-cholesterol diet markedly decreased the quantity of labeled cholesterol in the serum in both groups and obliterated the difference between control and lactating animals. This suggests that the increased hepatic cholesterol synthesis in the lactating animals is responsible for the differences in labeled cholesterol in the serum. Cholesterol feeding also reduced the quantity of labeled cholesterol localized to the mammary glands in lactating animals.(ABSTRACT TRUNCATED AT 250 WORDS)
Sodium and chloride transport across rabbit ileal brush border. II. Evidence for Cl-HCO3 exchange and mechanism of coupling
Authors: Knickelbein R. , Aronson P. S. , Schron C. M. , Seifter J. , Dobbins J. W.
Published: December 2017

An inside-alkaline pH gradient (pH 7.7 inside, 5.5 outside) stimulated Cl uptake in brush-border vesicles from rabbit ileum. The addition of HCO3 without changing the pH gradient further stimulated Cl uptake to a level fourfold greater than equilibrated Cl uptake. Although a K diffusion potential stimulated Cl uptake, this was insensitive to inhibition by 4,4-diisothiocyanostilbene-2,2'-disulfonate (DIDS), whereas pH and HCO3 gradient-stimulated Cl uptake was inhibited by DIDS. pH and HCO3 gradient-stimulated Cl uptake was found to be a saturable function of the Cl concentration with a Km of 3.5 mM and a Vmax of 49 nmol X mg prot-1 X min-1. To distinguish between coupling of Na and Cl transport by cotransport or dual exchange (Na-H and Cl-HCO3 exchange), we determined uptake with high (134 mM Tris-HEPES-MES) internal buffer and low (1.34 mM Tris-HEPES-MES) internal buffer concentrations. Inwardly directed gradients of 50 mM Na, 50 mM K, or 50 mM Na and K did not stimulate Cl uptake, and 50 mM Cl, 50 mM K, or 50 mM KCl did not stimulate Na uptake, with high internal buffer, excluding cotransport. In contrast, 50 mM Na stimulated Cl uptake (inhibited by 1 mM DIDS) and 50 mM Cl stimulated Na uptake (inhibited by 1 mM amiloride) in low buffer media. To determine a role for carbonic anhydrase, Na-stimulated Cl uptake was determined in low buffer media, equilibrated with either 100% N2 or 95% N2-5% CO2. Na stimulated Cl uptake 80% (compared with trimethylammonium control) with CO2 but only 30% with N2 (P less than 0.05). Acetazolamide partially inhibited (P less than 0.025) the stimulation of Cl uptake with CO2 but not with N2. Carbonic anhydrase activity was measured in homogenate and brush-border membrane and was enriched 7.9 +/- 0.4-fold, whereas sucrase was enriched 14.0 +/- 1.1-fold. We conclude that coupled Na and Cl transport occurs by dual exchange (Na-H and Cl-HCO3) and carbonic anhydrase, apparently located on the brush-border membrane, facilitates dual exchange by providing HCO3.
Possible mechanisms for the initiation and maintenance of postprandial intestinal hyperemia
Authors: Gallavan R. H. , Chou C. C.
Published: December 2017

Postprandial intestinal hyperemia is a locally mediated vascular response to the presence of nutrients in the lumen. In this review we discuss the role of various constituents of chyme in the development of the hyperemia and possible mechanisms of action. The luminal contents that produce the hyperemia are digested products of food; undigested food or pancreatic enzymes have no effect. Micellar fatty acids are the most potent vasodilators, whereas amino acids at physiological concentrations have little effect on intestinal blood flow. However, by-products of protein digestion are as potent as those of carbohydrates in increasing the blood flow. Bile increases ileal but does not alter jejunal blood flow. In addition, bile enhances the glucose-induced hyperemia and renders fatty and amino acids vasoactive. The mechanisms by which bile exerts its effect on the vasoactivity of these nutrients are poorly understood. The intestinal hyperemic response to the presence of nutrients in the lumen is mediated by a variety of regulatory pathways that vary with the nutrient. Factors involved include tissue metabolic rate, metabolites, nutrient absorption, tissue osmolality, tissue oxygen tension, intestinal peptides such as neurotensin and vasoactive intestinal polypeptide, and paracrine substances such as prostaglandins and histamine. It is likely that the hyperemia results from the complex interplay of all these factors on the intestinal vascular smooth muscle. Extrinsic and intrinsic nerves play a minor role in nutrient-induced hyperemia.
Methionine metabolism after portacaval shunt in the rat
Authors: Benjamin L. E. , Steele R. D.
Published: December 2017

The effect of portacaval shunt (PCS) on methionine metabolism in the rat was investigated. Male Sprague-Dawley rats were subjected to PCS and maintained on an 18% casein diet. Growth curves of operated rats were similar to controls. PCS rats excreted more urinary 35SO4 and less [35S]taurine than controls after intraperitoneal injection of 0.3 mmol/100 g [35S]methionine or [35S]cysteine. Total urinary taurine excretion was similar in PCS and control rats after a methionine or cysteine load; however, under basal conditions PCS rats had higher urinary taurine levels than controls, indicating that PCS may cause the taurine pool to be expanded. Hepatic methionine, S-adenosylmethionine, and cysteine pools were significantly decreased in PCS rats, while S-adenosylhomocysteine levels were unchanged. Relative rates of transsulfuration in PCS and control rats were studied by following the decrease in the 3H-to-35S ratio in liver protein after injection of [methyl-3H]methionine and [35S]methionine, and no difference in flux of 35S from [35S]methionine to [35S]cysteine was found. Similarly, total hepatic activities of methionine adenosyltransferase, cystathionine synthase, and cystathionine gamma-lyase were unchanged in PCS rats. These results indicate that altered methionine metabolism in PCS rats is not explained by changes in conversion of methionine to cysteine via the transsulfuration pathway.
Gastric mucosal barrier: stabilization of hydrophobic lining to the stomach by mucus
Authors: Hills B. A.
Published: December 2017

The hydrophobic nature [Am. J. Physiol. 244 (Gastrointest. Liver Physiol. 7): G561-G568, 1983] of the mucosal lining of canine oxyntic tissue has been characterized by rinsing with saline or a phospholipid solvent (hexane) and by measuring the maximum contact angles as droplets of a hydrophilic liquid (saline) and a hydrophobic liquid (olive oil) advance over that surface. The results are consistent with the luminal lining as an adsorbed monolayer of surfactant whose outer shell of packed fatty acid chains resemble polyethylene. Phospholipid (dipalmitoyl lecithin) monolayers deposited in vitro and polyethylene were both found to be readily wetted by stomach mucus, reducing advancing contact angles (theta) by 74 and 83%, respectively, while eliminating altogether the minimum contact angle for receding saline. Taking theta values for “dry” tissue as the most conservative, the reduction in surface energy imparted by mucus was estimated as 30.3 ergs X cm-2, which dispels the objection that, on thermodynamic grounds, surfactant must form a bilayer rather than a monolayer. Thus, as a powerful wetting agent, mucus would play an important role in stabilizing and replenishing the absorbed surfactant monolayer proposed as the physical basis for the gastric mucosal barrier.
Sodium dependence of luminal alkalinization by rabbit ileal mucosa
Authors: Smith P. L. , Cascairo M. A. , Sullivan S. K.
Published: December 2017

Stripped rabbit ileal mucosa was studied in vitro in Ussing chambers under short-circuit conditions using the pH-stat technique to determine basal rates of luminal alkalinization; the contribution of the shunt pathway to the alkalinization process; the effects of Na, Cl, or HCO3 removal from the bathing solutions on luminal alkalinization; and the effects of epinephrine, ouabain, 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS), acetazolamide, prostaglandin E1 (PGE1), A23187, sugars, or amino acids on the alkalinization process. Results from these studies reveal that, under basal conditions, the rate of luminal alkalinization accounts for 81% of the basal short-circuit current (Isc), although there was no correlation between the rate of alkalinization and Isc. The contribution of the shunt to the alkalinization process accounts for less than 10% of the mucosal-to-serosal HCO3 flux. Removal of Cl from the bathing solutions increased the rate of luminal alkalinization and decreased Isc. Sodium removal from the bathing solutions reduced both Isc and the rate of luminal alkalinization. Addition of DIDS to the luminal or serosal bathing solution reduced luminal alkalinization less than 30%. Acetazolamide, PGE1, and A23187 were all without effect on luminal alkalinization. Addition of 3-O-methyl-D-glucose or L-alanine to the luminal bathing solution did not alter luminal alkalinization but increased Isc, D-Glucose added to the luminal bathing solution reduced luminal alkalinization. This effect appears to result from metabolic acid production since 1) it is not seen with L-alanine or 3-O-methyl-D-glucose; 2) in the absence of HCO3 in the bathing solutions, D-glucose increased luminal acidification; and 3) luminal addition of fructose also reduced the rate of luminal alkalinization. Addition of epinephrine to the serosal bathing solution stimulates a Na-dependent serosal alkalinization process. These results suggest that luminal alkalinization results from Na-dependent, transcellular HCO3 transport and that a Na-dependent, HCO3 absorptive process is stimulated by adrenergic agents.
Migrating action potential complex: unmasked by 6-hydroxydopamine
Authors: Mathias J. R. , Clench M. H. , Davis R. H. , Sninsky C. A. , Pineiro-Carrero V. M.
Published: December 2017

We have previously described the myoelectric characteristics of a single moving ring contraction, the migrating action potential complex (MAPC), in rabbit ileal loops exposed to certain bacteria or their enterotoxins. The MAPC is thought to act as a defense mechanism of the host, clearing unwanted substances from the lumen. In the present study, 6-hydroxydopamine, a substance that selectively destroys adrenergic varicosities containing the neurotransmitter norepinephrine, unmasked the MAPC from the activity front of the migrating motor complex in an unanesthetized rat model. The animals developed diarrhea and lost weight. The study suggests that the MAPC may also be a physiological complex and under the modulation of the enteric nervous system. The MAPC may not be seen under normal control conditions because the complex migrates with the activity front and is under inhibitory control. Destroying the inhibitory mechanisms unmasked the MAPC from the activity front of the migrating motor complex and allowed neural transmission of the ring contraction.
Ileal mucosal growth during intraluminal infusion of ethylamine or putrescine
Authors: Seidel E. R. , Haddox M. K. , Johnson L. R.
Published: December 2017

Either ethylamine or the diamine putrescine was infused at the rate of 1 mumol/h for 66 h into the ileal lumen of rats. Total mucosal RNA, DNA, and protein content was greater in amine-treated rats than in rats receiving 0.9% NaCl. Growth was greatest in the mucosa surrounding the tip of the infusion catheter but was also observed 9 cm proximal and distal to the catheter tip. Infusion of these amines induced the activity of the enzymes ornithine and S-adenosylmethionine decarboxylase. Ornithine decarboxylase activity was increased 2- and 6-fold and S-adenosylmethionine decarboxylase activity 10- and 5-fold by putrescine and ethylamine, respectively. Induction of the polyamine biosynthetic enzymes was not accompanied by increases in the tissue content of polyamines. Putrescine, spermidine, and spermine content of the ileal mucosa surrounding the catheter tip was the same in 0.9% NaCl-, ethylamine-, and putrescine-treated animals. Finally, ethylamine was without effect on serum gastrin concentration in these experiments. The results suggest that amines regulate mucosal growth and may do so by modulating the activity of the enzymes involved in the synthesis of the polyamines.
Iron absorption in normal and iron-deficient beagle dogs: mucosal iron kinetics
Authors: Nathanson M. H. , Muir A. , McLaren G. D.
Published: December 2017

Absorption of dietary iron requires uptake of iron by the brush border of the intestinal epithelial cells, intracellular transport, and transfer to the systemic circulation. In iron-deficiency anemia, iron absorption is greatly increased, but the individual steps responsible for this increase have not been identified. We have developed a method to evaluate the rate constants for each of these steps, and we report here our results in beagle dogs a) under normal conditions and b) after phlebotomy to produce iron-deficiency anemia. Simultaneous administration of oral 59Fe3+-citrate and intravenous 55Fe-transferrin was used to investigate the kinetics of mucosal iron transport. Plasma levels of both isotopes and the whole-body excretion pattern of 59Fe were monitored sequentially, and the fractional mucosal transport rates were estimated by nonlinear least-squares fit of a physiologically based mathematical model to these data. Under normal conditions the fractional rate of mucosal iron uptake from the intestinal lumen was rate limiting, being less than 1% of the fractional rate of either iron incorporation into the mucosal storage pool or transfer of iron from the mucosa to the plasma. After induction of iron-deficiency anemia, the fractional mucosal iron uptake rate increased sixfold (P less than 0.005), while the rate of incorporation into the mucosal storage pool decreased ninefold (P less than 0.02); in contrast, the fractional rate of iron transfer to the plasma did not change. These results indicate that the enhanced iron absorption in iron-deficiency anemia is attributable to an increase in mucosal iron available for transfer to the plasma, leading in turn to a net increase in iron absorption, despite a normal fractional transfer rate.
Hypersensitive prostaglandin and thromboxane response to hormones in rabbit colitis
Authors: Zipser R. D. , Patterson J. B. , Kao H. W. , Hauser C. J. , Locke R.
Published: December 2017

Inflammation of the colon is associated with increased production of prostaglandins (PG) and thromboxanes (Tx), and these eicosanoids may contribute to the inflammatory, secretory, and motility dysfunctions in colitis. To evaluate the potential role of peptide hormones in the enhanced eicosanoid release, colitis was established in rabbits by a delayed-type hypersensitivity reaction to dinitrochlorobenzene and by an immune-complex-mediated reaction. PG and Tx were identified in the venous effluent of isolated perfused colons by radiochromatography after [14C]arachidonic acid prelabeling, as well as by bioassay, and then quantitated by immunoassay. The two colitis models were morphologically similar. Basal release of PGE2, PGI2, and TxA2 was two- to threefold greater from colitis tissue than from control tissue. Bradykinin (BK) and angiotensin II (ANG II) increased release of 14C-labeled eicosanoids, whereas several gastrointestinal hormones had no effect. In control colons, BK and ANG II increased PGE2 and PGI2 release (by about 2-fold) but did not alter TxA2. In contrast, BK and ANG II markedly exaggerated the release of eicosanoids in colitis. BK increased TxA2 release with 10-ng bolus injections in colitis, but there was no response with up to 10-micrograms bolus injections in control colons. The BK-induced Tx release in colitis was associated with an increase in vascular resistance (measured as perfusion pressure). Infusion of the selective Tx inhibitors dazoxiben and OKY-046 reduced TxB2 release by 96% and blunted the transient rise in perfusion pressure (from 17 +/- 5 to 5 +/- 2 mmHg). Since BK and possibly ANG II are increased at sites of inflammation, the hypersensitive eicosanoid response to these peptides may augment the eicosanoid-mediated manifestations of colitis.
Rat lingual lipase: effect of proteases, bile, and pH on enzyme stability
Authors: Roberts I. M.
Published: December 2017

In addition to initiating fat digestion in the stomach, lingual lipase may play a significant digestive role in the upper small intestine. By in vitro incubation techniques, the stability of rat lingual lipase at various physiological pH values, as well as the effects of pure proteases, rat gastric juice, bile, pancreatic juice, and mixed duodenal contents, on enzyme activity was explored. There were no changes in base-line activity of porcine pepsin, bovine carboxypeptidase-treated lipase, or heat-denatured proteases compared with controls after incubation at pH 2-6 at 37 degrees C for up to 1 h. In contrast, porcine trypsin-treated lipase demonstrated a significant loss from base-line activity to 59 +/- 12% (mean +/- SE) at pH 4, 34 +/- 11% at pH 6, and 41 +/- 4% at pH 8, and bovine chymotrypsin caused a loss in lipase activity to 11 +/- 7% at pH 8. Rat gastric juice containing 5,000 U pepsin reduced lipase activity to 17 +/- 5% of initial activity at pH 2 and to 45 +/- 3% at pH 4. Rat bile alone diminished activity only 35%, but rat pancreatic juice or mixed duodenal juice reduced lingual lipase activity to 1-12% of initial activity after 60 min at pH 6. Lingual lipase is particularly important in fat digestion in the stomach; however, its role in quantitative fat digestion under small intestinal conditions may be limited.
Regulation of fasting canine duodenal bile acid delivery by sphincter of Oddi and gallbladder
Authors: Scott R. B. , Eidt P. B. , Shaffer E. A.
Published: December 2017

Bile delivery into the duodenum during fasting is rhythmic and coordinated with the migrating myoelectric complex (MMC). To determine the role of the sphincter of Oddi (SO) and gallbladder (GB) in regulating duodenal bile acid delivery, three dogs were surgically prepared with a duodenal cannula and eight bipolar electrodes implanted from the duodenum to the terminal ileum. Fasting intestinal myoelectric activity was recorded during continuous intravenous infusion of [14C]taurocholic acid (TCA). Duodenal delivery of TCA was quantitated under four conditions: indirectly with the SO intact monitoring output with duodenal marker perfusion and directly with the SO cannulated and draining at three levels to vary outflow resistance (5 cm above, level with, or 20 cm below the SO). GB filling or emptying represented the algebraic difference between hepatic secretion (equal to the intravenous infusion rate) and rate of TCA delivery into the duodenum. With the sphincter intact the rate of bile acid delivery averaged one-half the intravenous infusion rate at 0-40% of the MMC cycle period, rose to exceed the intravenous infusion rate at 60-70% of the cycle period, and then fell to lower levels before the next MMC began. The pattern of delivery was identical with the SO cannulated, but the rate and percent infused TCA delivered into the duodenum depended significantly (P less than 0.01) on outflow resistance. They were least with the reservoir elevated 5 cm, intermediate when level, and greatest when 20 cm lower. Thus, duodenal bile acid delivery and GB filling or emptying during fasting were cyclically coordinated with the MMC. Peak rates of duodenal bile acid delivery and active GB emptying occur at 70% of the duodenal cycle period. Elimination of the SO does not eliminate the pulsatile pattern of fasting duodenal bile acid delivery, but outflow resistance determines the quantities delivered into the duodenum or stored in the GB. Thus, during the interdigestive period the net effect of SO function does not differ from a constant output resistance favoring the partition of hepatic biliary secretion into the GB and away from the duodenum, while the pulsatile pattern of duodenal bile acid delivery associated with the MMC results from cyclic active contraction of the GB.
Effect of glucose or fructose feeding on cholesterol synthesis in diabetic animals
Authors: Feingold K. R. , Moser A. H.
Published: December 2017

Previous studies have demonstrated that cholesterol synthesis is increased twofold in the small intestines of rats with streptozotocin-induced diabetes. The purpose of the present study was to determine the effect of adding glucose or fructose to standard rat chow on cholesterol synthesis in control and diabetic rats. In control rats a 25% glucose or fructose diet fed for 21 days markedly inhibited hepatic cholesterol synthesis in the liver. In contrast, in diabetic animals only fructose inhibited hepatic cholesterol synthesis. In both control and diabetic animals the addition of these simple sugars to the diet did not markedly alter extrahepatic cholesterol synthesis. The enhancement of small intestinal cholesterol synthesis observed in diabetic animals was present regardless of the dietary manipulations. Further studies demonstrated that the addition of smaller concentrations of fructose (10%) to standard rat chow decreased hepatic cholesterol synthesis in both control and diabetic rats. Similarly the addition of fructose to the diet of control and diabetics for a period as short as 2 days was also sufficient to inhibit hepatic cholesterol synthesis. In both control and diabetic animals, fructose feeding decreased hepatic 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity but did not alter the percentage of HMG-CoA reductase in the active form. Finally, the intestinal hypertrophy and stimulation of intestinal cholesterogenesis that are characteristic of streptozotocin-induced diabetes occurred when either glucose or fructose was the sole caloric source.