bluuebunny/crossref_metadata_2025 huggingface dataset:
Mucosal ornithine decarboxylase, polyamines, and hyperplasia in infected intestine
Authors: Wang J. Y. , Johnson L. R. , Tsai Y. H. , Castro G. A.
Published: December 2017

Ornithine decarboxylase (ODC), through the regulation of polyamine biosynthesis, influences normal mucosal growth and cell proliferation. The purpose of our study was to determine whether mucosal ODC activity and polyamines play a role in the dramatic increase in mucosal mass and crypt elongation associated with parasite-induced inflammation in the small intestine. Rats were inoculated orally with L1 larvae of the parasitic nematode Trichinella spiralis and killed at different times during the enteric phase of infection. Duodenal and jejunal mucosal ODC activities increased significantly from 2 to 14 days, peaking 7 days postinfection (PI). By 21 days PI, enzyme activity had returned to near normal values. In the ileal mucosa, ODC activity was increased only at 7 days PI. Increases in ODC activity were paralleled by increases in mucosal putrescine, spermidine, and spermine content. Infection with T. spiralis induced DNA synthesis and evoked a significant rise in DNA, RNA, and protein content in the mucosa. Increases in nucleic acid and protein levels were most prominent in the proximal half of the small intestine where the majority of worms reside. Treatment of rats with alpha-difluoromethylornithine (DFMO) prevented the infection-induced elevations in mucosal ODC activity, polyamine levels, DNA synthesis, and DNA, RNA, and protein content without influencing the development of inflammation or the parasite's life cycle. These results suggest that mucosal hyperplasia caused by infection may be regulated, in part, by the growth-promoting effects of ODC, presumably through the stimulation of crypt cell proliferation. Thus ODC may be an important determinant of the intestinal response to infection.
Comparison of primary and secondary esophageal peristalsis in humans: effect of atropine
Authors: Paterson W. G. , Hynna-Liepert T. T. , Selucky M.
Published: December 2017

To determine whether physiological differences exist between primary (swallow-induced) and secondary (distension-induced) peristalsis in humans, 10 healthy male volunteers underwent esophageal manometry on 2 consecutive days using a perfused intraluminal catheter system that incorporated a latex balloon. Initially the catheter was positioned so that the balloon was centered 16 cm above the lower esophageal sphincter (LES), and intraluminal pressures were recorded 21, 11, 6, and 1 cm above the LES. After a series of wet swallows, dry swallows, and balloon distensions, the catheter was repositioned so that the balloon was 6 cm above the LES and pressures were recorded 1 and 11 cm above the LES. A series of balloon distensions were repeated in this position, and the subject was then given either atropine (10 micrograms/kg iv) or placebo in a double-blind randomized fashion (on consecutive days). The protocol was then repeated in reverse order. Distension-induced responses aboral to the balloon with the balloon located 16 cm above the LES were 1) of lower amplitude, 2) more often nonperistaltic, and 3) less atropine sensitive than swallow-induced contractions at comparable sites. With the balloon located distally (6 cm above LES) contractions induced at the 11-cm site (i.e., orad to the balloon) were much more atropine sensitive than contractions induced at the same site when the balloon was located proximally (i.e., 16 cm above LES). These data suggest that, contrary to previous reports, secondary peristalsis differs significantly from primary peristalsis. Furthermore, atropine differentially effects these two types of peristalsis, suggesting that the neural pathways involved are dissimilar.
Localization of Y1 receptors for NPY and PYY on vascular smooth muscle cells in rat pancreas
Authors: Sheikh S. P. , Roach E. , Fuhlendorff J. , Williams J. A.
Published: December 2017

To localize binding sites for peptide YY (PYY) in the pancreas we utilized a slide-mount autoradiographic technique on frozen sections of rat pancreas incubated with 125I-Tyr36-PPY. Saturable autoradiographic labeling was located over pancreatic blood vessels, whereas islets, acinar cells, ducts, and neural elements did not appear to be specifically labeled. Isolated vascular fragments were prepared by collagenase digestion of rat pancreas. Binding experiments with 125I-Tyr36-PYY showed saturable binding to the fraction enriched in blood vessels but not to acini. Inhibition of 125I-Tyr36-PYY binding by nonradioactive neuropeptide Y (NPY) and PYY were similar, with half-maximal inhibition at 31.2 +/- 5 pM (n = 6); the potency of pancreatic polypeptide (PP) was 10,000 times lower. The binding site was classified as belonging to a Y1 type of NPY and/or PYY receptors, since [Leu31,Pro34]NPY, a specific Y1-receptor agonist, inhibited binding similar to NPY. To further localize the bound [125I-Tyr36]PYY within the blood vessels, light- and electron-microscopic autoradiographs were prepared and quantitated. Autoradiographic grains were located predominantly over vascular smooth muscle cells, although saturable localization was also seen over endothelial cells. It is concluded that in the pancreas Y1 receptors are predominantly located on vascular smooth muscle cells.
Ontogeny of epidermal growth factor receptor tyrosine kinase in rat liver
Authors: De B. K. , Brown T. L. , Suchy F. J.
Published: December 2017

The binding of epidermal growth factor (EGF) to its receptor and the activity of the receptor intrinsic protein-tyrosine kinase were studied during the ontogeny of rat liver. The number of EGF receptors during pre- and postnatal development was first compared in crude liver plasma membranes using 1) specific binding of 125I-labeled EGF and 2) immunoblot analysis using any antireceptor polyclonal rabbit antibody. Both methods detected the expression of the EGF receptor in fetal rat liver on day 17 of gestation, but in an amount markedly less than the adult. Within 24 h, there was a more than twofold increase in EGF binding to plasma membranes as well as a marked increase in receptor immunoreactivity. However, after birth, there was a precipitous drop in receptor number to less than 20% of the adult level by the end of the first postnatal day (P less than 0.001). Next, the presence of EGF-stimulated tyrosine kinase activity (autophosphorylation) was determined during the same stages of development. Electrophoresis of membranes phosphorylated in the presence or absence of EGF followed by autoradiography demonstrated autokinase activity stimulated by EGF in day 18 and 19 fetal liver plasma membranes, but not in membranes on day 17 of gestation. Similar to the pattern observed with EGF binding, there was a decrease in autokinase activity in early neonatal plasma membranes followed by an increase to near adult levels by 7 days postnatally. Quantitation of the amount of 32P radioactivity associated with the EGF receptor bands in each age group, correlated with the degree of autophosphorylation assessed by autoradiography.(ABSTRACT TRUNCATED AT 250 WORDS)
Intestinal lactase expression and epithelial cell transit in hormone-treated suckling rats
Authors: Yeh K. Y. , Yeh M. , Holt P. R.
Published: December 2017

Cortisone- and/or thyroxine (T4)-induced changes in jejunal lactase activity and epithelial cell migration were studied to determine the relationship of these two events. In suckling rats given a single dose of cortisone on day 6, jejunal lactase activity increased by 37% and cell turnover rate by 95% 3 days later, whereas T4 alone induced no changes. After cortisone plus T4, jejunal lactase activity decreased by 23% while cell turnover rate increased by 176%. Among all animal groups, the patterns of lactase expression along the crypt-villus (C-V) axis were similar, being low at the C-V junction, increasing to a high plateau at the mid- or third quarter villus level, and decreasing slightly at the villus tip. The calculated epithelial cell age at half maximum lactase expression in cortisone-treated and cortisone plus T4-treated rats was 30 and 53% younger than in control rats. Maximum lactase activity in villus cells was approximately 45% higher in cortisone-treated than in control or cortisone plus T4-treated rats. Parallel measurements of sucrase and lactase activities along C-V axis showed elevated lactase activity at higher villus positions than lead sucrase activity, suggesting that cortisone action occurs in villus cells. Thus jejunal lactase expression may be modulated by 1) adjusting villus cell age required for maximum expression, 2) altering the level of lactase activity in villus cells, and 3) changing the turnover rate of lactase containing epithelial cells.
Role of blood flow and alkaline secretion in acid-induced deep duodenal villous injury in rats
Authors: Leung F. W.
Published: December 2017

The effect of 16,16-dimethylprostaglandin E2 (dmPGE2) and corticotropin-releasing factor (CRF) on duodenal blood flow, alkaline secretion, and acid-induced deep duodenal villous injury was studied. The duodena of anesthetized rats were prepared for simultaneous measurement of alkaline secretion by back titration, and blood flow by hydrogen gas clearance; or for perfusion with 0.1 N HCl and histological examination of villous injury. The results revealed that the dmPGE2-induced increase in basal alkaline secretion (due solely to an increase in the volume of secretion) appears to be a better predictor of protection against exogenous acid-induced deep duodenal villous injury than rise in duodenal blood flow, since CRF induces a similar rise in duodenal blood flow but does not enhance alkaline secretion or reduce acid-induced villous damage. The absence of a greater loss of H+ during acid perfusion of the duodenum in the dmPGE2-treated rats, however, suggests that the mechanism of the dmPGE2 protection against acid-induced deep duodenal villous injury cannot be explained entirely by its ability to increase basal duodenal alkaline secretion.
Expression of cobalamin transport proteins and cobalamin transcytosis by colon adenocarcinoma cells
Authors: Ramanujam K. S. , Seetharam S. , Ramasamy M. , Seetharam B.
Published: December 2017

Human colon adenocarcinoma (Caco-2) cells express both intrinsic factor-cobalamin receptor and transcobalamin II (TC II). The expression of these activities began to rise by day 6 and reached peak levels between 10 and 15 days in culture. The postconfluent Caco-2 cell membranes bound approximately 30-35 fmol of intrinsic factor (IF) [57Co]Cbl/mg protein. The size of the mature receptor expressed in the apical brush border had a relative molecular mass of 230 kDa. The intracellular form of TC II had a Mr of 43, 5 higher than the secreted form of TC II. TC II was secreted unidirectionally via the basolateral direction when Caco-2 cells were grown on culture inserts. When grown on culture inserts, the Caco-2 cells were polarized (electrical resistance greater than 200 omega/cm2) and transcytosed [57Co]Cbl bound to IF from apical-to-basal but not from basal-to-apical direction. Under these conditions, [57Co]Cbl complexed to haptocorrin was not transported. These cells also transcytosed free [57Co]Cbl, although less efficiently. The [57Co]Cbl transcytosed using either IF[57Co]Cbl or free [57Co]Cbl as ligands was bound exclusively to TC II. Intracellular [57Co]Cbl decreased during transcytosis with a slow (t1/2 = 4 h) transfer of [57Co]Cbl from IF to TC II. These results show that the transport of Cbl in Caco-2 cells is very similar to the human enterocyte system.
Is the small intestinal epithelium truly "tight" to inulin permeation?
Authors: Ma T. Y. , Hollander D. , Erickson R. A. , Truong H. , Krugliak P.
Published: December 2017

In this study, we evaluated the "leakiness" of intestinal epithelium through examination of small intestinal absorption of inulin in vivo by perfusing rat jejunum with 10 microM inulin. In physiological conditions, we found significant absorption of inulin at a rate of 44.6 nmol.100 cm-1.h-1 or absorption of 14.7%.100 cm-1.h-1 of the amount perfused. Increasing water flux by changing the luminal osmolarity resulted in linear (y = 31.1 + 2.4x, r = 0.97) increase in absorption of inulin, indicating a significant convective component of inulin absorption. There was large permeation of inulin at net water secretion and at zero net water fluxes (31.1 nmol.100 cm-1.h-1), indicating significant absorption of inulin by diffusive movement as well. The small intestinal tissue retention of inulin occurred rapidly within the first 15 min of perfusion, and the total tissue retention remained unchanged thereafter at approximately 10.8 nmol/100 cm. 16,16-Dimethylprostaglandin E2 decreased water flux, whereas cyclooxygenase inhibitors, indomethacin and acetylsalicylate, increased water flux. Inulin absorption closely paralleled changes in water flux induced by these agents. Taurocholate also caused parallel decrease in water and inulin absorption. Varying the resistance of unstirred water layer with changing luminal flow rate, the addition of mucolytic agent acetylcysteine, or alterations of luminal pH did not affect water or inulin absorption. We conclude that inulin permeates the small intestinal epithelium in significant amounts under normal physiological conditions, presumably through the paracellular pathways utilizing aqueous channels.
Actions of peptide YY and neuropeptide Y on chief cells from guinea pig stomach
Authors: Raufman J. P. , Singh L.
Published: December 2017

Peptide YY (PYY), found in intestinal endocrine cells, and neuropeptide Y (NPY), a structural analogue of PYY found in neurons, inhibit gastric, pancreatic, and intestinal fluid and electrolyte secretion. We examined the effects of these peptides on dispersed chief cells from guinea pig stomach. PYY and NPY, but not pancreatic polypeptide, starting at nanomolar concentrations, caused a 40–50% inhibition of secretin-, vasoactive intestinal polypeptide-, prostaglandin E2-, and forskolin-induced increases in chief cell adenosine 3',5'-cyclic monophosphate (cAMP) content and pepsinogen secretion. These inhibitory peptides did not alter pepsinogen secretion caused by cholecystokinin, carbamylcholine, A23187, 8-bromo-cAMP, or a phorbol ester. The inhibitory effects of PYY on chief cell cAMP production occurred within 30 s, were independent of phosphodiesterase activity, and did not affect the actions of cholera toxin. However, the inhibitory effects of PYY were abolished when chief cells were preincubated with pertussis toxin, an agent that uncouples inhibitory guanine nucleotide binding (G) proteins from their receptors. In gastric chief cells, PYY and NPY attenuate the stimulatory effects of secretagogues whose actions are mediated by changes in cellular levels of cAMP. PYY-induced attenuation of chief cell adenylate cyclase activity appears to involve activation of inhibitory G proteins.
Bicarbonate-induced activation of taurocholate transport across the basal plasma membrane of human term trophoblast
Authors: el-Mir M. Y. , Eleno N. , Serrano M. A. , Bravo P. , Marin J. J.
Published: December 2017

The efflux of [14C]taurocholate from previously loaded vesicles, obtained from basal plasma membrane of human trophoblast, was studied. Apparent Km (620 microM) and Vmax (1.79 nmol.min-1.mg protein-1) values were similar to those found in influx experiments (Marin et al., Gastroenterology 99: 1431-1438, 1990). Transmembrane gradients of both bicarbonate (100 mM) and unlabeled taurocholate (0.5 mM) accelerated [14C]taurocholate efflux. The bicarbonate-induced effect was not abolished by carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and K(+)-valinomycin voltage clamp. Neither was it mimicked by 5,5'-dimethyloxazolidine 2,4-dione (DMO) or by other organic (taurine, glycine, lactate, or acetate) or inorganic (Cl-, SCN-, HPO24-, or SO24-) anions, and it was not sensitive to carbonic anhydrase inhibitors. No effect of bicarbonate was observed either in the absence of gradient or in the presence of a cis-directed gradient. Bicarbonate-induced transstimulation was related to an increase in the value for the apparent Vmax (+30%). Study of the stoichiometry suggests that the most probable coupling ratio is one, bicarbonate: taurocholate. In summary, these results provide evidence for the existence of a bicarbonate-driven anion exchange in the basal plasma membrane of the human term placental trophoblast.
Effect of muscle length on isometric stress and myosin light chain phosphorylation in gallbladder smooth muscle
Authors: Washabau R. J. , Wang M. B. , Dorst C. L. , Ryan J. P.
Published: December 2017

These experiments were designed to characterize the effect of muscle length on isometric stress, sensitivity to stimulation, and phosphorylation of the 20,000-Da myosin light chains in guinea pig gallbladder smooth muscle. Basal, active, and total isometric stress were determined in acetylcholine- or K(+)-treated (10(-4) M ACh, 80 mM KCl) muscle strips at 0.6-1.3 times the optimal muscle length (Lo) for isometric stress development. The effect of muscle length on the sensitivity to ACh and K+ was determined in cumulative dose-response experiments (10(-8) to 10(-4) M ACh, 10-80 mM KCl) at 0.7, 1.0, and 1.3 Lo. The effect of muscle length on myosin light chain phosphorylation was determined in ACh- or K(+)-treated (10(-4) M ACh, 80 mM KCl) muscle strips at 0.7, 1.0, and 1.3 Lo. In gallbladder smooth muscle, 1) active isometric stresses at 0.7 and 1.3 Lo were less than active isometric stress at 1.0 Lo; 2) the sensitivity of developed stress was similar at 1.0 and 1.3 Lo but decreased at 0.7 Lo; 3) the decline in isometric stress and sensitivity at 0.7 Lo was associated with reduced levels of phosphorylated myosin light chain; and 4) the decline in isometric stress at 1.3 Lo was not associated with reduced amounts of phosphorylated myosin light chain. These results suggest that the decline in active stress and sensitivity at short muscle lengths (L less than Lo) in gallbladder smooth muscle is due, at least in part, to decreases in the activation of the myofilaments. The decline in active isometric stress at long muscle lengths (L greater than Lo) is not due to changes in myofilament activation.
Extent and modulation of junctional communication between pancreatic acinar cells in vivo
Authors: Chanson M. , Orci L. , Meda P.
Published: December 2017

To assess whether junctional communication may be of physiological relevance in the control of exocrine pancreas secretion, we have studied acinar cell coupling by microinjecting Lucifer Yellow CH in the intact pancreas of anesthetized rats. Reconstructions from serial sections showed that, under control conditions, pancreatic cells are extensively coupled within each acinus but do not communicate with centroacinar cells, duct cells, and cells of neighboring acini. Intravenous infusion of acetylcholine and caerulein, or electrical stimulation of the vagus nerve, increased pancreatic secretion (P less than 0.02-0.001). Under these stimulatory conditions, the extent of acinar cell communication was decreased (P less than 0.001) by 40%. The acetylcholine-induced uncoupling was prevented by treating rats with atropine. Thus, in the intact pancreas, acinar cells intercommunicate extensively within each acinus under resting conditions and reduce their coupling during stimulation. These data support the view that modulation of cell coupling is a physiologically relevant mechanism for the regulation of exocrine pancreas secretion in vivo.
Protective actions of a leukotriene B4 antagonist in splanchnic ischemia and reperfusion in rats
Authors: Karasawa A. , Guo J. P. , Ma X. L. , Tsao P. S. , Lefer A. M.
Published: December 2017

Pentobarbital-anesthetized rats were subjected to occlusion of both the celiac and superior mesenteric arteries for 90 min followed by reperfusion for 2 h. All seven rats given only the vehicle died within 2 h of reperfusion, whereas rats treated with LY-255283 (3 or 10 mg/kg iv), a leukotriene B4 (LTB4) receptor antagonist given 10 min before reperfusion, exhibited significantly higher survival rates of 57% (4 out of 7) and 75% (6 out of 8), respectively, 2 h after reperfusion. Rats given 1 mg/kg of LY-255283 showed no significant improvement in survival. Splanchnic artery occlusion (SAO)-shock rats treated with LY-255283 (3 or 10 mg/kg) exhibited significantly attenuated accumulation of plasma free amino-nitrogenous compounds and of a myocardial depressant factor. Treatment with LY-255283 (10 mg/kg) markedly (P less than 0.01) ameliorated the deficits of endothelium-dependent relaxation of isolated superior mesenteric artery (SMA) rings in untreated SAO-shock rats. LY-255283 at 10 mg/kg significantly attenuated the increased myeloperoxidase activity in the intestinal tissue of SAO-shock rats. Moreover, LY-189444, a closely related compound having no LTB4 antagonist activity, did not protect rats in SAO shock, whereas a lipoxygenase inhibitor confirmed protection in SAO shock. These results suggest that LTB4 plays a pivotal role in endothelial dysfunction occurring in SAO-shock rats by chemoattraction and activation of neutrophils on the surface of vascular endothelial cells. Moreover, LY-255283 but not LY-189444 inhibited the adherence of rat neutrophils to isolated SMA endothelium.(ABSTRACT TRUNCATED AT 250 WORDS)
Increased glucose uptake by intestinal mucosa and muscularis in hypermetabolic sepsis
Authors: Lang C. H. , Obih J. C. , Bagby G. J. , Bagwell J. N. , Spitzer J. J.
Published: December 2017

The purpose of the present study was to determine the following: 1) whether the sepsis-induced increase in glucose uptake was a generalized response along the entire length of the gastrointestinal tract; 2) the relative contribution of the mucosa and muscularis to the enhanced uptake; and 3) whether reducing intestinal blood flow would attenuate the elevated rate of glucose uptake. Hypermetabolic sepsis increased in vivo glucose uptake in all sections of the gastrointestinal tract (57-93%) except the stomach. The rates of glucose uptake per gram of tissue by the mucosa and muscularis were not different. However, because the mucosa accounted for the majority of the whole intestine mass, this layer was responsible for 76-78% of the glucose uptake by the entire small intestine. Intestinal blood flow, determined with the use of radiolabeled microspheres, increased by 127% in sepsis. In both groups, approximately 70% of the total intestinal blood flow was distributed to the mucosa. Somatostatin was infused to produce splanchnic vasoconstriction and decreased the sepsis-induced increment in intestinal flow to the mucosa and muscularis (38 and 54%), whereas the enhanced rate of glucose uptake was not altered. Somatostatin also produced a severe insulinopenia. These results indicate that hypermetabolic sepsis increases glucose uptake to a similar extent along the length of the small and large intestine and that the majority of this increase is due to an enhanced uptake by the mucosa.(ABSTRACT TRUNCATED AT 250 WORDS)
Myogenic oscillatory mechanism for opossum esophageal smooth muscle contractions
Authors: Helm J. F. , Bro S. L. , Dodds W. J. , Sarna S. K. , Hoffmann R. G. , Arndorfer R. C.
Published: December 2017

We evaluated the control of phasic contractions in opossum esophageal circular smooth muscle by determining the contractile response in vitro to agents that cause membrane depolarization and excitation by different mechanisms. Transverse muscle strips taken from different sites along the length of the smooth muscle esophagus were exposed to progressively increasing concentrations of tetraethylammonium (1-30 mM), K+ (4.6-30 mM), or bethanechol (10(-6) to 10(-2) M). In normally inactive esophageal circular smooth muscle, tetraethylammonium and high K+ concentration elicited phasic contractions that were not blocked by atropine and tetrodotoxin. Bethanechol, an M2 muscarinic receptor agonist that acts selectively on smooth muscle, elicited phasic contractions that were not blocked by tetrodotoxin. We conclude that a latent myogenic oscillatory mechanism for control of phasic contractions exists in esophageal circular smooth muscle and that it may be activated by nonspecific excitation of the smooth muscle membrane. We suggest that this myogenic oscillatory mechanism is likely excited and modulated by nerves.
Inhibitory effect of ileal oleate on postprandial motility of the upper gut
Authors: Dreznik Z. , Brocksmith D. , Meininger T. A. , Soper N. J.
Published: December 2017

To determine the effect of ileal oleate on postprandial gastrointestinal motility, duodenal and paired perfusion-aspiration ileal catheters and bipolar duodenal and jejunal electrodes were surgically implanted in five dogs. The ileum was perfused with either saline or an isotonic oleic acid emulsion at 2 ml/min. A 205-kcal mixed meal containing 120 ml liquid nutrient labeled with 111In-diethylenetriamine pentaacetic acid (DTPA) and solid food labeled with 99mTc was then administered orally. Gastric emptying was assessed by a gamma camera, myoelectric activity was continuously monitored, and duodenal-ileal transit of phenol red was determined over the ensuing 240 min. Ileal oleate reduced duodenal spikeburst frequency by 50% (P less than 0.05) and delayed gastric emptying of liquids and solids. Four hours after ingesting the meal, 62% of solids and 34% of liquids were retained in the stomach during oleic acid perfusion compared with 25 and 4%, respectively, when saline was perfused (P less than 0.05). Duodenal-ileal transit was markedly slowed by ileal perfusion with the oleic acid emulsion (P less than 0.001). Ileal oleate therefore exerted a profound inhibitory effect on proximal gut motility in the early period after ingestion of a mixed-nutrient meal in dogs.
Effects of epidermal growth factor on diamine oxidase expression and cell growth in Caco-2 cells
Authors: Daniele B. , Quaroni A.
Published: December 2017

To investigate the role of diamine oxidase (DAO) in the intestinal mucosa, we compared its expression with cell proliferation and differentiation in the human colon carcinoma cell line Caco-2. DAO synthesis was evaluated in subconfluent and confluent cultures and in the presence of epidermal growth factor (EGF), a polypeptide hormone known to have specific trophic effects on the small intestinal mucosa. EGF stimulated DNA synthesis, significantly increased cellular DAO activity and the amount of enzyme secreted into the culture medium, but decreased expression of dipeptidyl peptidase IV, a marker of cell differentiation in confluent Caco-2 cells. Immunoprecipitation of DAO from cells labeled metabolically with [35S]methionine failed to demonstrate an increased enzyme synthesis in EGF-treated cells, suggesting that this hormone acted primarily at a posttranslational level by reducing DAO degradation before intracellular storage or secretion. A possible relationship between changes in cellular DAO activity and cell proliferation was also investigated by using aminoguanidine, a specific and potent DAO inhibitor. Although DAO activity was markedly suppressed, aminoguanidine had no significant effects on the rate of DNA synthesis. These results demonstrated that in Caco-2 cells EGF stimulated DNA synthesis and DAO expression; however, cell proliferation and differentiation were not correlated with the levels of cellular DAO, suggesting that this enzyme does not play a major role in the regulation of intestinal epithelial cell turnover.
Inhibition of activity-induced parotid growth by submandibular gland ablation or autonomic denervation
Authors: Schneyer C. A. , Humphreys-Beher M. G. , Hall H. D.
Published: December 2017

When solid chow was reintroduced for 2 days to rats previously maintained on an all-liquid diet, [3H]thymidine incorporation into rat parotid gland was 2.4-4.4 times that of chow-fed or liquid-fed animals. When the submandibular-sublingual glands were removed before the change in diet from liquid to solid food, values for [3H]thymidine incorporation of parotid gland were not statistically different from those of chow-fed rats: the increase was completely prevented. The increase in gland size that accompanied the change in dietary consistency was not prevented by prior submandibular gland ablation; [3H]uridine incorporation was also not different from that of chow controls. The removal of both the sympathetic and parasympathetic nerves to the parotid gland before the dietary manipulation also suppressed [3H]thymidine incorporation into parotid, and values did not differ from chow controls. Moreover, the return to normal gland size (chow level) also was prevented by autonomic denervation. The data show an important role of the submandibular gland in regulation of the thymidine increase associated with the enhanced parotid activity that accompanies the change from liquid to solid diet. However, the submandibular gland does not have an important role in regulation of the increase in gland size that also accompanies the dietary change, whereas the autonomic nerves have an important role in regulation of hypertrophy and hyperplasia of parotid gland.
Liver denervation does not alter the circadian rhythm of bile acid synthesis in rats
Authors: Gilberstadt M. L. , Bellinger L. L. , Lindblad S. , Duane W. C.
Published: December 2017

In both rats and humans there is a distinct circadian rhythm of bile acid synthesis that is independent of feedback regulation. To determine whether the circadian rhythm is directly mediated via hepatic nerves, bile acid synthesis was studied in selectively liver-denervated male Sprague-Dawley rats in a bile fistula model. Complete denervation was confirmed by histofluorescent staining for neural elements in frozen sections of livers. There was no significant difference in mean bile acid synthesis, amplitude of the circadian rhythm, or time of peak synthesis between the denervated rats and nondenervated controls. In one denervated rat studied four times at weekly intervals, there was no shift in acrophase, indicating that the rhythm had not become free running. We conclude that signals arriving via hepatic nerves neither directly cause nor entrain the circadian rhythm of bile acid synthesis in rats.
An ontogenically regulated 48-kDa protein is a component of the Na(+)-bile acid cotransporter of rat liver
Authors: Ananthanarayanan M. , Bucuvalas J. C. , Shneider B. L. , Sippel C. J. , Suchy F. J.
Published: December 2017

Recent evidence suggests that the Na(+)-coupled carrier mechanism for bile acids on the hepatocyte basolateral plasma membrane is a polypeptide in the molecular weight range of 48,000-50,000. In this study we used a strategy for the identification and isolation of this transport protein based on the observation that Na(+)-dependent transport activity is abruptly expressed in fetal rat liver just before birth [Suchy et al. Am. J. Physiol. 251 (Gastrointest. Liver Physiol. 14): G665-G673, 1986]. Analysis of basolateral plasma membranes by SDS-PAGE revealed that a protein of apparent molecular weight 48,000 was absent from fetal rat liver on day 19 of gestation, barely detectable on day 20, and thereafter increased progressively with postnatal development. Monospecific, polyclonal antibodies raised against the 48-kDa protein but not preimmune antibodies significantly inhibited the initial rate of Na(+)-dependent taurocholate uptake by isolated rat hepatocytes. In contrast, Na(+)-independent taurocholate transport and uptake of another anion, 35SO4(2-), were not affected by antibody treatment. When an extract containing the total complement of basolateral proteins was incorporated into asolectin liposomes, Na+ gradient-dependent uptake of taurocholate was observed, including a 2- to 2.5-fold accumulation of substrate above its equilibrium concentration (overshoot). However, if the membrane extract was first selectively depleted of the 48-kDa protein by immunoprecipitation with the anti-48-kDa antibody before reconstitution, Na(+)-dependent stimulation of taurocholate transport was completely abolished. These studies indicate that an ontogenically regulated 48-kDa protein is a component of the basolateral Na(+)-dependent transport system for bile acids.
Neural 5-hydroxytryptamine receptors regulate chloride secretion in guinea pig distal colon
Authors: Cooke H. J. , Wang Y. Z. , Frieling T. , Wood J. D.
Published: December 2017

The effects of 5-hydroxytryptamine (5-HT) on epithelial short-circuit current (Isc) were determined and related to the 5-HT effects on electrical and synaptic behavior of neurons in the submucosal plexus of the guinea pig colon. 5-HT evoked a biphasic increase in Isc that was reduced by bumetanide, Cl(-)-free solutions, atropine, and mecamylamine and abolished by tetrodotoxin. The 5-HT response was mimicked by 2-methyl-5-HT, but not by 5-hydroxyindalpine, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane, and 5-methoxytryptamine (5-MeOT). ICS 205-930 suppressed the 5-HT response. Electrical field stimulation of submucosal neurons evoked an increase in Isc indicative of Cl- secretion that was reduced by 5-MeOT and enhanced by 2-methyl-5-HT. Application of 5-HT to submucosal neurons by micropressure ejection resulted in membrane depolarization, augmented excitability, and repetitive spike discharge. The depolarization was biphasic, consisting of rapidly and slowly activating components. The rapidly activating component was suppressed by ICS 205-930. Fast excitatory postsynaptic potentials evoked by electrical stimulation of interganglionic connectives were suppressed by 5-HT and 5-MeOT. These results suggest that 5-HT activates 5-HT3 receptors, which mediate fast excitatory responses in submucosal neurons, leading to release of acetylcholine at nicotinic and muscarinic synapses and stimulation of Cl- secretion. Presynaptic inhibition suppresses acetylcholine release and results in attenuation of neurally evoked Cl- secretion.
Hepatocellular 22Na+ uptake: effect of oleate
Authors: Sorrentino D. , Van Ness K. , Moukabary K. , Berk P. D.
Published: December 2017

Whether cellular oleate uptake is Na+ coupled remains controversial. Our present studies document that hepatocellular [3H]oleate uptake is unaltered by isomotic substitution of Na+ with K+, Li+, or sucrose in Hanks' HEPES buffer. In parallel studies 22Na+ uptake was significantly (P less than 0.02) increased by concentrations of alanine that yielded [3H]-alanine uptakes greater than 24% of basal 22Na+ uptake when both were expressed as nanomoles per minute per 10(6) hepatocytes. Although [3H]glutamine uptake exceeded this threshold, maximal specific [3H]taurocholate uptake did not. Consistent with the observations with alanine, addition of glutamine, but not taurocholate, to the incubations resulted in a significant increase in 22Na+ uptake. The ionophore monensin increased uptake of 22Na+ under all conditions. Both in the absence and in the presence of HCO3- (4 and 25 mM), specific [3H]oleate uptake was sufficient to elicit a readily detectable effect on 22Na+ uptake, if Na+ and oleate were cotransported. However, addition of 1 mM oleate did not affect 22Na+ uptake in the absence of HCO3- (5.4 +/- 0.6 vs. control 5.4 +/- 0.4 nmol.min-1.10(6) cells-1) as well as in the presence of 4mM HCO3- (10.4 +/- 1.8 vs. control 9.9 +/- 1.1) and 25 mM HCO3- (10 +/- 1.3 vs. control 10.8 +/- 0.5). These data indicate that the predominant component of hepatocellular oleate uptake is not directly associated with Na+ influx. They do not exclude, however, a more complex or indirect link.
Phosphoinositide metabolism in intestinal smooth muscle: preferential production of Ins(1,4,5)P3 in circular muscle cells
Authors: Murthy K. S. , Makhlouf G. M.
Published: December 2017

The pattern of inositol phospholipid metabolism in response to stimulation by contractile agonists was examined in muscle cells isolated separately from circular and longitudinal muscle layers of guinea pig intestine. Addition of cholecystokinin octapeptide (CCK-8) to circular muscle cells caused a prompt decrease in phosphatidylinositol 4,5-bisphosphate (PtdInsP2) (50 +/- 6%) and PtdInsP (38 +/- 9%), which reached a nadir in 15 s. Addition of CCK-8 to longitudinal muscle cells caused a decrease in PtdInsP only (42 +/- 6%); PtdInsP2 levels, which were three times lower in this cell type, did not change throughout the period of stimulation. Hydrolysis of PtdInsP2 in circular muscle cells was accompanied by a concentration-dependent increase in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]; the maximal increase was 10 to 16 times greater in circular muscle cells as measured by radioreceptor assay and by ion-exchange chromatography. The small amounts of InsP3 produced in longitudinal muscle cells did not result from more rapid degradation, since the rates of disappearance of exogenous Ins(1,4,5)P3 in both cell types were similar. Within 5 s after addition of CCK-8, InsP3 comprised 73 +/- 5% of total inositol phosphates produced in circular muscle cells and 2.0 +/- 0.2% in longitudinal muscle cells. These divergent patterns indicate that inositol phospholipid metabolism is channeled toward generation of InsP3 in circular muscle cells only; the metabolic pattern in these cells parallels the selective presence of high-affinity InsP3 receptors and the pattern of intracellular Ca2+ mobilization.
Myosin light chain phosphorylation and contraction of guinea pig gallbladder smooth muscle
Authors: Washabau R. J. , Wang M. B. , Ryan J. P.
Published: December 2017

These experiments were designed to determine 1) whether acetylcholine (ACh) stimulation is accompanied by changes in myosin light chain phosphorylation in gallbladder smooth muscle and 2) whether dephosphorylated noncycling cross bridges (latch bridges) exist in gallbladder smooth muscle. Isometric stress, isotonic shortening velocity, and myosin light chain phosphorylation were determined under conditions of contraction and relaxation in ACh-stimulated guinea pig gallbladder smooth muscle. Unstimulated muscle contained 6.8 +/- 2.0% phosphorylated myosin light chain. ACh stimulation (5 x 10(-5) or 10(-4) M) was associated with a rapid increase in myosin light chain phosphorylation to a value that was maintained throughout the tonic contraction. In contrast, isotonic shortening velocity was maximal at 30 s of stimulation and then declined over time to a steady-state level that was 25-30% of the peak velocity. Upon agonist washout (relaxation), dephosphorylation of the myosin light chain occurred at about the same rate as the decline in shortening velocity and preceded the decline in isometric stress. These data suggest that ACh stimulation is accompanied by changes in myosin light chain phosphorylation but that dephosphorylation of cross bridges is not necessary for the slowing of cross-bridge cycling rates in gallbladder smooth muscle.
Transport of 5-methyltetrahydrofolate in basolateral membrane vesicles of rat liver
Authors: Horne D. W. , Reed K. A. , Said H. M.
Published: December 2017

We examined 5-methyltetrahydrofolate transport across the basolateral membrane (BLM) of rat liver using isolated membrane vesicles. Uptake of 5-methyltetrahydrofolate by BLM vesicles (BLMV) was found by osmolarity and initial uptake studies to be mostly the result of transport of the substrate into an active, intravesicular space with some binding (approximately 25%) to membrane surfaces. Uptake was similar in the presence of an inwardly directed Na+ or K+ gradient, indicating that transport is not dependent on Na+. Uptake of 5-methyltetrahydrofolate was linear for the first 30 s and was more rapid when an initial pH gradient was imposed across the vesicular membrane [extravesicular pH (pHo) = 5.0, intravesicular pH (pHi) = 7.5]. Under pH gradient conditions, uptake at 3-5 min was about twofold higher than at equilibrium (60 min). The initial rate of uptake at pHo = pHi = 5.0 was more rapid than at pHo = pHi = 7.5, but in neither case was uptake above equilibrium observed. Uptake under pH gradient conditions and at pH 5.0 (no pH gradient) was saturable (apparent Michaelis constants of 0.58 and 0.36 microM, respectively; maximum uptake rates of 1.25 and 0.79 pmol.10 s-1.mg protein-1, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)
Ontogeny of membrane and soluble amino-oligopeptidases in rat intestine
Authors: Reisenauer A. M. , Lee E. A. , Castillo R. O.
Published: December 2017

Intestinal amino-oligopeptidase (AOP) plays an essential role in protein digestion. To characterize its postnatal development, we measured AOP activity in intestinal membrane and cytosolic fractions in suckling and weaned rats, compared the subunit structures of the membrane and soluble enzymes, and assessed the biochemical relationship of these peptidases. At weaning, jejunal membrane AOP activity doubled while soluble AOP activity in the ileum fell abruptly. The maturational increase in the molecular mass of ileal membrane AOP was due to alterations in the N-linked glycosylation of this protein. Ileal membrane and soluble AOP exhibited similar substrate affinities, pH optima, inhibition characteristics, and antigenic epitopes. However, soluble AOP was 25-35 kDa smaller than the membrane enzyme. Peak incorporation of [35S]methionine into ileal brush-border AOP preceded maximal radioactivity in soluble AOP, suggesting that the membrane peptidase is a precursor of the soluble enzyme. We conclude that membrane and soluble AOP are closely related proteins with distinct developmental profiles and that the soluble peptidase may be derived from endocytosis of the membrane enzyme.
Absorption and lymphatic transport of peroxidized lipids by rat small intestine in vivo: role of mucosal GSH
Authors: Aw T. Y. , Williams M. W. , Gray L.
Published: December 2017

The absorption and lymphatic transport of peroxidized MaxEPA fish oil was studied using the lymph fistula rat to determine the role of mucosal glutathione (GSH) in intestinal metabolism of luminal lipid hydroperoxides. Decreasing intestinal GSH concentrations with buthionine sulfoximine (BSO, 1.15 +/- 0.20 nmol/g), diethyl maleate (DEM, 0.93 +/- 0.26 nmol/g), phorone (1.46 +/- 0.14 nmol/g), or 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, 1.54 +/- 0.18 nmol/g) compared with control (2.60 +/- 0.38 nmol/g) resulted in higher luminal recovery of the infused lipid hydroperoxide (% of infused dose): BSO (87.8 +/- 4.8%), DEM (86.1 +/- 1.3%), phorone (78.1 +/- 2.1%), and BCNU (71.7 +/- 4.8%) compared with control (52.8 +/- 4.3%). These results suggest that decreased elimination of luminal peroxidized lipids is associated with decreased tissue GSH. Treatment of rats with BSO, DEM, phorone, or BCNU resulted in dramatic increases in appearance of peroxidized lipids in lymph over 6-h lipid infusion (54.7 +/- 3.7, 57.7 +/- 4.6, 46.4 +/- 2.7, and 42.1 +/- 3.9 nmol, respectively) compared with control (20.5 +/- 3.4 nmol). The results are consistent with decreased intracellular metabolism of absorbed hydroperoxides and enhanced transport into lymph under GSH-deficient conditions. The current findings suggest that the function of the mucosal GSH peroxidase/oxidized glutathione (GSSG) reductase system may play an important role in intestinal handling of luminal lipid hydroperoxides. A compromised function of this detoxication mechanism in GSH-deficient states can significantly alter the metabolic fate of dietary peroxidized lipids.
EndothelinB receptor on guinea pig small intestinal smooth muscle cells
Authors: Yoshinaga M. , Chijiiwa Y. , Misawa T. , Harada N. , Nawata H.
Published: December 2017

We investigated the binding characteristics of the endothelin (ET) receptor and the mechanism by which ET induces contraction of longitudinal smooth muscle cells of the guinea pig small intestine by using vasoactive intestinal contractor (VIC), a mouse variant of ET-2. A functional receptor for VIC was found to exist on longitudinal smooth muscle cells. These cells showed a similar binding of and contractile response to ET-1, ET-2, and ET-3. Inhibitors of both intracellular and extracellular Ca2+ movement attenuated the VIC-induced contraction of longitudinal smooth muscle cells. These results suggest that smooth muscle cells of the guinea pig small intestine express the ETB receptor that primarily mediates the contractile effect on smooth muscle cells. In addition, ET-induced contraction depends on intracellular as well as extracellular Ca2+.
Differential intestinal deconjugation of taurine and glycine bile acid N-acyl amidates in rats
Authors: Zhang R. , Barnes S. , Diasio R. B.
Published: December 2017

Mechanisms responsible for the difference in the relative amounts of taurine- and glycine-conjugated bile acid N-acyl amidates (Tau/Gly ratio) are not fully understood. In the present study, the stability of taurine- and glycine-conjugated bile acid N-acyl amidates during intestinal transit and absorption was examined to investigate the contribution of intestinal deconjugation to the Tau/Gly ratio in rat bile. Radiolabeled chenodeoxycholic acid (CDC) and its N-acyl amidates with glycine (CDC-Gly) or taurine (CDC-Tau) were introduced into the lumen of the upper small intestine in the biliary fistula rats, and radioactive metabolites in bile, blood, urine, and tissues were identified and quantitated by high-performance liquid chromatography. Results indicated that 1) extensive deconjugation of CDC-Gly occurs during intestinal absorption; 2) CDC-Tau is recovered in bile largely intact; and 3) newly synthesized CDC-Tau and CDC-Gly are formed in a ratio of less than 2:1 after administration of [14C]-CDC. In summary, the present study demonstrates that resistance of taurine-conjugated bile acid N-acyl amidates to hydrolysis in the intestine, rather than a difference in synthesis of taurine- and glycine-conjugated N-acyl amidates in liver, may account for the high Tau/Gly ratio in rat bile.
Transforming growth factor-beta (TGF-beta 1) inhibits pancreatic acinar cell growth
Authors: Logsdon C. D. , Keyes L. , Beauchamp R. D.
Published: December 2017

Effects of transforming growth factor (TGF)-beta 1 on mouse pancreatic acinar cell growth and rapid intracellular responses to cholecystokinin (CCK) were examined in vitro. TGF-beta 1 inhibited [3H]thymidine incorporation stimulated by either the CCK analogue caerulein, epidermal growth factor, or insulin. TGF-beta 1 inhibition of growth stimulated by a maximal dose of caerulein (1 nM) was dose dependent with one-half maximal effects occurring at approximately 5 pM and maximal inhibition seen with 30 pM. In contrast to its effects on CCK-stimulated [3H]thymidine incorporation, TGF-beta 1 had no effect on CCK-stimulated increases in amylase release or intracellular Ca2+ concentration. To determine whether TGF-beta 1 might be an autocrine growth regulator, pancreatic mRNA was probed for the presence of TGF-beta 1 transcripts. TGF-beta 1 mRNA was not detected in whole pancreas but was detectable with increasing abundance over time in primary cultures of pancreatic acinar cells. The appearance of the TGF-beta 1 mRNA corresponded to the period of rapid cellular proliferation in vitro. These results suggest that TGF-beta 1 may be an autocrine growth inhibitor in the pancreas and that the inhibitory effects of TGF-beta 1 on pancreatic acinar cell growth occur at sites distal to those involved in stimulus-secretion coupling.