2-(Anilinomethyl)imidazolines as alpha(1)-adrenoceptor agonists: the identification of alpha(1A) subtype selective 2'-carboxylic acid esters and amides.

2-(Anilinomethyl)imidazolines with 2'-esters or 2'-amides are potent agonists of the cloned human alpha(1)-adrenoceptors in vitro. The size and shape of the ortho substituent can have significant effects on the potency, efficacy, and subtype selectivity of these 2-(anilinomethyl)imidazolines. alpha(1A)-subtype selective agonists have been identified.

Endogenous prostanoids control ion transport across neonatal porcine ileum in vitro.

In contrast to the net absorption of Na and Cl ions observed in vivo, porcine small intestine had a net secretion of these ions in vitro. These discrepancies between in vivo and in vitro results have led to difficulties in interpretation of studies investigating mechanisms of intestinal secretion and diarrhea in this species. To examine the influence of endogenous prostanoids on ion transport in neonatal porcine ileum in vitro, tissues were prepared and studied in indomethacin. Net absorption of Na, reversal of net Cl secretion to net absorption, and decreased short circuit current were observed. Conversely, addition of prostaglandins to indomethacin-treated tissues reversed these effects and reestablished conditions similar to those observed in control tissues. Control tissue was essentially refractory to the effects of exogenous prostaglandins. Results indicate that under in vitro conditions, ion transport in neonatal porcine ileum is tightly regulated by endogenous prostanoids that abolish the neutral NaCl absorptive mechanism and elicit electrogenic Cl secretion. However, concentrations of these prostanoids may have been artificially high as a result of tissue preparation for in vitro study.

Villous atrophy, crypt hyperplasia, cellular infiltration, and impaired glucose-Na absorption in enteric cryptosporidiosis of pigs.

Intestinal morphology and fluid and electrolyte transport were examined in a neonatal porcine model of cryptosporidiosis. Sections of jejunum, ileum, and colon were obtained for morphometric analysis on days 3, 6, 9, and 12 postinfection, and in vivo perfusion studies of jejunum and ileum were conducted on days 3 and 4 postinfection. The most severe morphologic lesion was seen in the ileum on day 3, and consisted of villous atrophy, crypt hyperplasia, and cellular infiltration. Villous surface area was reduced from 2.1 +/- 0.4 x 10(5) microns2 in control ileum to 0.8 +/- 0.1 x 10(5) microns2 in infected ileum, a result associated with enterocytes that were fewer in number and reduced in cross-sectional area. Conversely, the number of inflammatory cells in the lamina propria of the villus increased from 456 +/- 116 in control to 1014 +/- 187 in infected villus without a significant change in the volume of the lamina propria. At the height of infection, there was an approximate 1:2 ratio of both organisms and inflammatory cells to villous enterocytes. In contrast, organisms were not observed in the crypts, and the concentration of inflammatory cells in crypt lamina propria was unaltered. Disappearance of organisms and polymorphonuclear cells from the ileum was associated with restoration of normal structure and was complete by day 12. Although organisms were seen in the colon, the general architecture was not severely affected. On days 3 and 4 postinfection, there was a complete impairment of the glucose-stimulated Na and water absorption in both jejunum and ileum of infected pigs; however, absorption of electrolytes and water from a basic Ringer's solution, in the absence of glucose, was not significantly affected. These results are consistent with a malabsorptive diarrheal disease associated with the morphological damage and are very similar to those seen in enteric viral disease in pigs, except that the upper intestine is more severely affected in the latter.

Morphologic and functional effects of bile salt on the porcine colon during injury and repair.

Deoxycholate-induced colonic injury and repair were studied both functionally and morphologically utilizing in vivo loop preparations of the porcine colon. The mucosa was exposed to (a) varying doses (1.5 to 21 mM) of deoxycholate for 30 minutes, (b) 15 mM deoxycholate for varying times and (c) 15 mM deoxycholate for 30 minutes with varying times of recovery. Colonic permeability was assessed by mannitol clearance from blood to lumen and transmural potential difference. After colonic perfusion, tissue samples were collected for light and electron microscopy. Both the degree of mucosal permeability and the amount of superficial epithelial damage increased with increasing concentrations of bile salt culminating in cell necrosis and epithelial sloughing. Denuded colonic surfaces became reepithelialized by migrating, flattened cells in as little as 15 minutes of recovery. Relatively normal appearing columnar epithelium was restituted within 2 hours. Mannitol clearance returned to control values after 30 minutes of recovery, whereas it took potential difference 2 hours of recovery to return to normal. The results of these experiments suggest that (a) the permeability changes measured are most likely due to the lytic action of bile salts which leads to cell degeneration and sloughing of the superficial epithelium, (b) epithelial restitution after superficial damage is remarkably rapid, (c) the formation of a flattened epithelium of immature cells is adequate for restoration of the barrier to macromolecules but ion transport or resistance is slower to recover and (d) repair is due to an active ameboid movement of viable cells out of the crypts onto the surface of the colon.

Effect of prostaglandin inhibitors on bile salt-induced mucosal damage of porcine colon.

The effect of endogenous prostaglandin inhibition on bile salt-induced colonic injury and secretion was studied microscopically and by measurements of [14C]mannitol clearance and transmural potential difference in vivo. Bile salt-induced mucosal damage and permeability increased sequentially with concentration, and these degenerative changes were accelerated with the cyclooxygenase inhibitor indomethacin. Mepacrine, a phospholipase inhibitor, gave similar results, whereas nordihydroguaiaretic acid, a lipoxygenase inhibitor, was ineffective. The effect of indomethacin was abolished by prostaglandin E2 replacement; however, exogenous prostaglandin or prior bile salt exposure failed to result in additional protection. Concentrations of bile salts below the threshold for damage elicited net secretion in the presence or absence of indomethacin, and indomethacin was also without effect on the bile salt-induced secretion at damaging concentrations. Restitution of a completely denuded surface was unaffected by indomethacin, and occurred within 30 min of recovery. The present evidence suggests that endogenous prostaglandins render the mucosa more resistant to acute injury by events independent of the repair process. In addition, the bile salt-induced secretion, which can be dissociated from increased mucosal permeability and microscopic damage, is unlikely to be the result of increased mucosal synthesis of prostaglandins.

Restitution of barrier and transport function of porcine colon after acute mucosal injury.

Acute injury of the porcine colonic epithelium was induced in vivo with the bile salt, deoxycholate. A concentration of 15 mM for 30 min completely destroyed the surface epithelium and induced a marked increase in mucosal permeability to mannitol. The crypt epithelium however was not significantly affected. Within 8 min of recovery, the colonic surface was reepithelialized with flattened, migrating cells, and within 40 min, mucosal permeability to mannitol was normalized. In vitro studies showed that in these early stages of recovery, NaCl transport, short-circuit current, and resistance were markedly impaired, whereas the theophylline-induced secretory response remained intact. Recovery of absorptive function paralleled the transition from flattened to columnar surface epithelium and was complete within 2 h. Results suggest that 1) active migratory events play an important role in rapid restitution of an epithelial barrier, 2) active absorption of ions is much slower to recover, and 3) active secretory events are intact and probably originate in the crypt epithelium.

Intestinal oxalate-degrading bacteria reduce oxalate absorption and toxicity in guinea pigs.

Previous studies have provided evidence that an anaerobic bacterium, which degrades dietary oxalate to CO2 and formate, is present in colonic contents of a number of herbivorous species, laboratory rodents and humans. The present study examines the possibility that these bacteria degrade significant amounts of oxalate and can influence colonic oxalate absorption. Guinea pigs adapted to a diet containing 2% sodium oxalate or fed a normal diet were challenged with 67, 135, 170 or 200 mg of sodium oxalate containing 0.5 microCi of [14C]oxalate, which was injected into the cecum. Adapted animals excreted approximately 2% of the 14C in the urine, regardless of the dose, whereas unadapted animals excreted significantly higher amounts in the urine at the two lower doses and died at the two higher doses. Conversely, antibiotic treatment of adapted guinea pigs reduced the ability of their cecal flora to degrade oxalate, and a correspondingly greater percentage of an injected oxalate load was excreted in the urine. Oxalate degradation rates in cecal fluid were depressed by the secondary bile salt deoxycholate, and in vitro studies with pure isolates of guinea pig and human strains of oxalate degraders confirmed that these bacteria were highly sensitive to low concentrations of deoxycholate. Results indicate that these bacteria may be important in preventing excess absorption of oxalate and raise the possibility that the hyperoxaluria associated with bile salt malabsorption of ileal disease in part may be due to suppression of these bacteria by the bile salts.

Effects of slaframine on ruminant digestive function: liquid turnover rate and fermentation patterns in sheep and cattle.

Two trials were initiated to determine if slaframine (SF) can be used to alter fluid digesta flow and fermentation patterns in the rumen. In trial 1, a preliminary experiment, four Dorset X Barbados Black-belly ruminal-cannulated wethers (avg weight 41.6 8.7 kg) given ad libitum access to a pelleted concentrate/hay diet were injected intramuscularly with 0, 12, 24 or 48 micrograms SF/kg body weight (BW) in a 4 X 4 Latin-square design. Ruminal fluid dilution rate was determined using a single intraruminal infusion of polyethylene glycol (7 g), followed by seven hourly ruminal fluid samples. The administration of 48 micrograms SF/kg BW increased (P less than .10) ruminal volume and outflow by 27 and 25%, respectively, compared with controls. In trial 2, two Hereford and two Angus ruminal cannulated steers (avg weight 568 +/- 93 kg) were injected with 0, 6, 12 or 24 micrograms SF/kg BW at 8-h intervals over a 24-h period in a 4 X 4 Latin-square design. Steers were fed a concentrate diet at twice maintenance in 24 equal portions daily. Ruminal fluid dilution was measured using a single intraruminal infusion of cobalt-ethylenediamine tetraacetic acid (20 g) administered 9 h after the initial SF injection. Ruminal fluid was collected each hour during 8 to 24 h after the initial SF injection and analyzed for pH, osmolality and volatile fatty acids (VFA).(ABSTRACT TRUNCATED AT 250 WORDS)