Mechanisms underlying ACh induced modulation of neurogenic and applied ATP constrictions in the submucosal arterioles of the guinea-pig small intestine.
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1. Role of the vascular endothelium in acetylcholine (ACh) induced modulation of neurogenic and applied ATP (adenosine 5'-triphosphate) constrictions of intestinal submucosal arterioles was investigated. 2. Arteriole constrictions, induced either by exogenous ATP or evoked by perivascular nerve stimulation, were attenuated in the presence of ACh. 100 nM ACh almost completely abolished neurogenic constrictions whereas up to 10 microM ACh reduced constrictions to exogenous ATP by only about 60%. 3. Treatment of the arterioles with 100 microM Nomega-nitro-L-arginine (NOLA) and 5 microM indomethacin, to block respectively nitric oxide (NO) and prostanoid release from the endothelium, had no effect on the ACh induced inhibition of neurogenic constrictions but significantly attenuated the inhibitory effects of ACh on constrictions to exogenous ATP. 4. Disruption of the vascular endothelium had no effect on the ACh induced inhibition of neurogenic constrictions but attenuated the inhibitory effects of ACh on applied ATP constrictions to the same extent as after treatment with NOLA and indomethacin. In comparison, endothelial disruption completely abolished the inhibitory effect of substance P (SP) on exogenously applied ATP constrictions. 5. 50 nM ACh significantly attenuated the amplitude of neurally evoked excitatory junction potentials (ejps) recorded from the vascular smooth muscle without altering the time constant of decay (taudecay) of the ejps. 6. It is concluded that ACh inhibits neurogenic constrictions by prejunctional modulation of transmitter release from the perivascular sympathetic nerves with no major role for endothelial paracrine factors. 7. Endothelial NO and/or prostanoids mediate some of the ACh induced inhibition of constrictions to exogenous ATP whereas the endothelium independent inhibitory effects of ACh are attributed to a direct action of ACh on the vascular smooth muscle. However, an indirect effect resulting from activation of vasodilator nerves cannot be ruled out. (+info)
Nutritional evaluation of poultry by-product meal as a protein source for ruminants: small intestinal amino acid flow and disappearance in steers.
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Six Angus steers (260+/-4 kg initial BW) fitted with ruminal, duodenal, and ileal cannulas were used in a 6 x 6 Latin square design to evaluate the effect of feeding poultry by-product meal (PBM) on small intestinal flow and disappearance of amino acids. The diets were provided at 2% of BW on a DM basis, formulated to contain 11.5% CP, and consisted of 49% corn silage, 36% cottonseed hulls, and 15% supplement on a DM basis. Supplements were formulated to contain 37% CP with sources of supplemental N being soybean meal (100% SBM) and 0, 25, 50, 75, and 100% PBM, with urea used to balance for N. Duodenal flow of all amino acids increased linearly (P < .07) as PBM increased in the diet and, except for His, increased (P < .09) for 100% PBM compared with 100% SBM. Similar results were observed for duodenal flow of nonbacterial amino acids, which linearly increased (P < .05) with PBM and were greater (P < .05) for 100% PBM than for 100% SBM. Soybean meal increased (P < .09) the duodenal flow of nonbacterial Lys compared with 0% PBM, and 0% PBM increased (P < .04) flow of Val, Ala, and Pro compared with 100% SBM. Duodenal bacterial essential, nonessential, and total amino acid flows were not affected (P > .80) by PBM; however, they were greater (P < .02) for 100% SBM than for 100% PBM. In addition, nonessential and total bacterial amino acid flows were increased (P < .06) for 100% SBM compared with 0% PBM. Small intestinal disappearance of Lys and Pro increased linearly (P < .09) as PBM increased, and 100% PBM increased (P < .07) disappearance of Arg and Ala compared with 100% SBM. Supplemental N source had no effect (P > .31) on apparent small intestinal disappearance of essential, nonessential, and total amino acids. These data suggest that when PBM, SBM, and urea were used as sources of supplemental N, the daily disappearance of amino acids from the small intestine of steer calves consuming a corn silage- and cottonseed hull-based diet was similar. (+info)
Receptor-mediated vascular and metabolic actions of endothelin-1 in canine small intestine.
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The effects of endothelin-1 (ET-1) infusion on blood flow (QG) and O2 uptake (VO2G) were examined in the small intestine of anesthetized dogs (n = 10). Arterial and venous flows of a gut segment were isolated, and the segment was perfused at constant pressure. Arterial and gut venous blood samples were taken, gut perfusion pressure and QG were measured, and O2 extraction ratio (OERG) and VO2G were calculated. ET-1 was infused (0.118 microgram. kg-1. min-1 ia) throughout the experiment. In group 1 (n = 5), ETA receptors were blocked using BQ-123 (0.143 mg. kg-1. min-1 ia) followed by blockade of ETB receptors with BQ-788 (0.145 mg. kg-1. min-1 ia). The order of ETA and ETB receptor blockade was reversed in group 2 (n = 5). In group 1, the decrease in QG observed with ET-1 infusion was partially reversed with BQ-123; no further change occurred after BQ-788 administration. In group 2, addition of BQ-788 to the infusate further decreased QG, whereas addition of BQ-123 returned QG to a value not different from that with ET-1 infusion alone. These data indicated that ET-1-induced vasoconstriction in the gut was mediated via ETA receptors and that this constriction was buffered by activation of ETB receptors. VO2G decreased in proportion to the decrease in QG with ET-1, decreased further with ET-1 plus ETB receptor blockade (group 2), and increased in proportion to the increases in QG with ETA receptor blockade (both groups). No changes in OERG occurred during ETA and ETB receptor antagonism in either group. This study is the first to demonstrate that a flow-limited decrease in gut VO2G occurred with infusion of ET-1 in gut vasculature. An intriguing and novel finding was that, during O2 limitation, OERG was only 50% of that normally associated with ischemia in this tissue. (+info)
The influence of nitric oxide (NO) on adenosine-induced metabolic effects was studied in the intestine. Blood flow supplied an in situ- isolated segment of small intestine in anesthetized cats via the superior mesenteric artery (SMA) and was controlled by a vascular circuit. The SMA and portal samples were taken for analysis of oxygen and lactate. Adenosine (0.4 mg. kg-1. min-1, intra-SMA) reduced oxygen consumption by 25.1 +/- 2.9 from 73.1 +/- 10.8 micromol. min-1. 100 g-1 and increased lactate production by 13.3 +/- 3.0 from 12.8 +/- 4.6 micromol. min-1. 100 g tissue-1 during constant-flow (CF, decreased shear stress) but not during constant-pressure (CP, increased shear stress) perfusion. Blockade of NO synthase using Nomega-nitro-L-arginine methyl ester did not affect the metabolic effects of adenosine during CF but eliminated the differences seen between CP and CF perfusion. A NO donor, 3-morpholinosydnonimine, attenuated the metabolic effects of adenosine during CF perfusion. The results suggested that shear-induced NO antagonized metabolic effects of adenosine but that the inhibition of vascular effects by NO was not shear dependent since it occurred in both CP and CF perfusion. (+info)
Gastrointestinal responses to a panel of lectins in rats maintained on total parenteral nutrition.
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Total parenteral nutrition (TPN) causes atrophy of gastrointestinal epithelia, so we asked whether lectins that stimulate epithelial proliferation can reverse this effect of TPN. Two lectins stimulate pancreatic proliferation by releasing CCK, so we asked whether lectins that stimulate gastrointestinal proliferation also release hormones that might mediate their effects. Six rats per group received continuous infusion of TPN and a once daily bolus dose of purified lectin (25 mg. rat-1. day-1) or vehicle alone (control group) for 4 days via an intragastric cannula. Proliferation rates were estimated by metaphase arrest, and hormones were measured by RIAs. Phytohemagglutinin (PHA) increased proliferation by 90% in the gastric fundus (P < 0.05), doubled proliferation in the small intestine (P < 0.001), and had a small effect in the midcolon (P < 0.05). Peanut agglutinin (PNA) had a minor trophic effect in the proximal small intestine (P < 0.05) and increased proliferation by 166% in the proximal colon (P < 0.001) and by 40% in the midcolon (P < 0.001). PNA elevated circulating gastrin and CCK by 97 (P < 0.05) and 81% (P < 0.01), respectively, and PHA elevated plasma enteroglucagon by 69% and CCK by 60% (both P < 0.05). Only wheat germ agglutinin increased the release of glucagon-like peptide-1 by 100% (P < 0.05). PHA and PNA consistently reverse the fall in gastrointestinal and pancreatic growth associated with TPN in rats. Both lectins stimulated the release of specific hormones that may have been responsible for the trophic effects. It is suggested that lectins could be used to prevent gastrointestinal atrophy during TPN. Their hormone-releasing effects might be involved. (+info)
Cloning and functional expression of an SGLT-1-like protein from the Xenopus laevis intestine.
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A cDNA encoding an Na+-glucose cotransporter type 1 (SGLT-1)-like protein was cloned from the Xenopus laevis intestine by the 5'- and 3'-rapid amplification of cDNA ends method. The deduced amino acid sequence was 673 residues long, with a predicted mass of 74.1 kDa and 52-53% identity to mammalian SGLT-1s. This gene was expressed in the small intestine and kidney, reflecting a tissue distribution similar to that of SGLT-1. The function of the protein was studied using the two-microelectrode voltage-clamp technique after injection of cRNA into Xenopus laevis oocytes. Perfusion with myo-inositol elicited about twofold larger inward currents than perfusion with D-glucose. The order of the substrate specificity was myo-inositol > D-glucose > D-galactose >/= alpha-methyl-D-glucoside. The current induced by myo-inositol increased with membrane hyperpolarization and depended on external myo-inositol and Na+: the apparent Michaelis-Menten constant was 0.25 +/- 0.07 (SD) mM with myo-inositol, whereas the apparent concentration for half-maximal activation was 12.5 +/- 1.0 mM and the Hill coefficient was 1.6 +/- 0.1 with Na+. In conclusion, the cloned protein shares features with both SGLT-1 and the Na+-myo-inositol cotransporter. (+info)
Quantitative analysis of peristalsis in the guinea-pig small intestine using spatio-temporal maps.
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1. Peristalsis was evoked in guinea-pig small intestine by slow fluid infusion and recorded onto video and digitized. Spatio-temporal maps of diameter and longitudinal movement were constructed and parameters of motion were calculated. 2. During the filling of the isolated segments of intestine, rhythmic local longitudinal movements were observed at several points along the preparation. These phasic longitudinal muscle contractions were associated with small but significant local increases in diameter and probably reflect a passive mechanical coupling by connective tissue in the gut wall. In addition, occasional synchronized longitudinal muscle contractions caused net shortening of the preparation and always preceded the onset of peristaltic emptying. 3. Peristaltic emptying was characterized by a contraction of the circular muscle which usually started at the oral end of the preparation, that propagated aborally, propelling the contents. However, in 19 % of trials, the first circular muscle contraction occurred in the aboral half of the preparation. 4. The propagation of peristalsis consisted of separate sequential circular muscle contractions several centimetres long, particularly in the oral half of the preparation, giving a 'step-like' appearance to the spatio-temporal map. The gut was transiently distended aboral to the propagating circular muscle contraction due to the propulsion of contents. 5. At each point in the preparation, the longitudinal muscle remained contracted during the propulsive part of the circular muscle contraction. Only when the circular muscle contraction became lumen occlusive did lengthening of the longitudinal muscle take place. 6. Spatio-temporal maps are a powerful tool to visualize and analyse the complexity of gastrointestinal motility patterns. (+info)
Dysregulation of beta-catenin is common in canine sporadic colorectal tumors.
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Human colorectal tumorigenesis is often initiated by APC (adenomatous polyposis coli) or beta-catenin (CTNNB1) mutations, which result in dysregulation of beta-catenin expression, followed by alterations in E-cadherin and/or p53. We examined 32 canine intestinal tumors for expression and intracellular distribution of beta-catenin, E-cadherin, and p53 using immunohistochemistry. beta-Catenin in normal mucosal epithelial cells was restricted to lateral cell membranes, but 13/13 (100%) colorectal adenomas had intense cytoplasmic and/or nuclear reactivity. Three of six (50%) colorectal carcinomas, 2/13 (15%) small intestinal carcinomas, and dysplastic cells in 1/2 focal hyperplastic lesions in the small intestine had a similar pattern of staining; remaining tumors had normal membranous beta-catenin reactivity. There was a correlation (P = 0.007) between abnormal beta-catenin and E-cadherin staining with 11/13 (85%) colorectal adenomas, 3/6 (50%) colorectal carcinomas, and 3/13 (23%) small intestinal carcinomas showing decreased membranous reactivity compared with normal mucosal epithelium. E-cadherin staining was reduced more often in adenomas than in carcinomas (P = 0.04). There were two patterns of nuclear p53 staining: > 60% of nuclei in 2/26 (8%) carcinomas (one colorectal, one small intestinal) were strongly labeled, whereas three colorectal adenomas and one small intestinal carcinoma had fainter staining in 10-20% of cells. Dysregulation of beta-catenin appears to be as important in canine colorectal tumorigenesis as it is in the human disease and could be due to analogous mutations. Malignant progression in canine intestinal tumors does not appear to be dependent on loss of E-cadherin or beta-catenin expression or strongly associated with overexpression of nuclear CMI antibody-reactivity p53. (+info)