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To define a potential role for the angiotensin system in Crohn's colitis, the colonic mucosal levels of angiotensin I and II were measured in endoscopic biopsy samples from patients with active Crohn's colitis (n = 20), ulcerative colitis (n = 13), other forms of colitis (n = 3), and normal controls (n = 17). Colonic mucosal levels of angiotensin I and II were greater in patients with Crohn's colitis than in normal subjects (p less than 0.001 and p less than 0.001, respectively). Mucosal levels of angiotensin I and II were also higher in Crohn's colitis than in ulcerative colitis (p less than 0.001 and p less than 0.001, respectively), and levels of angiotensin II were higher in Crohn's than in other forms of colitis (p = 0.014). Mucosal levels of angiotensin I and II correlated well with the degree of macroscopic inflammation in Crohn's colitis (r = 0.86, p less than 0.001 and r = 0.68, p less than 0.001, respectively). Mucosal levels of angiotensin I correlated fairly well with the Crohn's Disease Activity Index (r = 0.46, p less than 0.05) while angiotensin II levels correlated poorly. These studies suggest that angiotensin I and II may have a role in the inflammation associated with Crohn's colitis.

Interleukin 1 or 3 added to the serosal side of chicken small intestine transiently increases short-circuit current. Replacement of bathing-medium Cl and HCO3 with gluconate and HEPES abolished the short-circuit current increase, consistent with these cytokines stimulating electrogenic anion secretion. Cytokine-stimulated short-circuit current changes were inhibited by preincubation with piroxicam (10(-5) M), an inhibitor of arachidonic acid cyclooxygenase, suggesting prostaglandin formation as an intermediate step for cytokine stimulation of short-circuit current. In intact mucosal strips, interleukin 1 and 3 stimulated prostaglandin E2 release and elevated tissue 3',5'-cyclic adenosine monophosphate concentration. When prostaglandin E2 release from epithelial and subepithelial fractions of the mucosa by interleukin 1 was determined, increases were found only from the subepithelium. The secretory actions of cytokines appear to be mediated by arachidonic acid metabolites most likely produced by cells of the lamina propria and submucosa and may play a role in inflammatory processes in which intestinal secretion is enhanced.

Previous studies in the ferret demonstrated that vagal nerve stimulation induced an atropine-resistant water secretion. Substance P and vasoactive intestinal polypeptide are possible mediators of this secretory response. The objectives of this study were to investigate the in vivo effects of substance P and vasoactive intestinal polypeptide on the jejunal musculature and epithelium. Substance P caused an increase in jejunal motility, water secretion, and transmural potential difference. Cholinergic blockade did not affect the substance P-induced contractions, but did reduce the increase in transmural potential difference, suggesting an inhibition of water secretion. Vasoactive intestinal polypeptide abolished motor activity; however, it induced an increase in transmural potential difference that was atropine and tetrodotoxin resistant. By immunohistochemical methods, immunoreactive vasoactive intestinal polypeptide and immunoreactive substance P were localized to both nerve cell bodies and nerve fibers in the ferret intestine. Determination of intestinal concentrations of vasoactive intestinal polypeptide and substance P in the ferret showed concentrations of these two neuropeptides that were similar to those in human intestine and demonstrated much higher concentrations of these substances in the muscular layer than in the epithelial layer. Our data demonstrate that in the ferret substance P excites and vasoactive intestinal polypeptide inhibits jejunal motor activity. However, both peptides increase water secretion. Our results suggest that in response to vagal stimulation, neuronally released substance P or vasoactive intestinal polypeptide may participate in the atropine-resistant water secretion.

The presence of receptors for insulinlike growth factor I and II in human colonic epithelium is demonstrated. Scatchard analysis of binding data obtained with 125I-insulinlike growth factor I showed an insulinlike growth factor I receptor with a dissociation constant of 8.6 nM and a binding capacity of 0.8 pmol/mg membrane protein. Distinct insulinlike growth factor II receptors labeled with 125I-insulinlike growth factor II were also found with a dissociation constant of 6.9 nM and a binding capacity of 4.7 pmol/mg membrane protein. Two sets of observations make it possible to discriminate between the two types of insulinlike growth factor receptors. (a) Unlabeled insulinlike growth factor I was 3 times more potent than insulinlike growth factor II in inhibiting [125I] insulinlike growth factor I binding. Conversely, unlabeled insulinlike growth factor II was 10 times more potent than insulinlike growth factor I in competing with 125I-insulinlike growth factor II for binding to membranes. Insulin and proinsulin did not compete with either of the tracers. (b) Affinity labeling of membranes followed by sodium dodecylsulfate polyacrylamide gel electrophoresis under reducing conditions, revealed a radio-ligand-receptor complex of molecular weight 130,000 and 250,000 using 125I-insulinlike growth factor I and 125I-insulinlike growth factor II, respectively. These observations indicate that adult human colonic epithelium is abundantly equipped with two sets of receptors that recognize preferentially either insulinlike growth factor I or insulinlike growth factor II.

Existing methods for the histochemical demonstration of gastrointestinal cells are somewhat limited. Chromogranin represents a family of proteins that coexist with catecholamines in the secretory vesicles of adrenal medulla cells. In the present study, immunocytochemistry was used to test whether chromogranin is a marker for gut endocrine cells. Serial sections of each area of human gut were immunostained for chromogranin and for the amine and each of the peptides known to be present in mucosal endocrine cells. Chromogranin was immunostained in large numbers of endocrine cells in all tissues examined. All identified endocrine cell types were found, in serial sections or by sequential silver impregnations, to be chromogranin immunoreactive. However, the possibility exists that some chromogranin-immunoreactive cells contain a yet to be discovered endocrine substance. Immunostaining of chromogranin thus appears to provide a means for demonstrating all gastrointestinal mucosal endocrine cells identifiable by the methods described in this study.

During fat absorption, active synthesis of cholesterol, phospholipids, and specific apolipoproteins are required for chylomicron formation and secretion. In the inherited disease abetalipoproteinema, chylomicrons cannot be made in response to fat feeding, and they as well as low and very low density lipoproteins are completely absent from plasma. The genetic defect in the disease is presumed to be an inability to synthesize apolipoprotein B, the apoprotein common to all the above lipoprotein classes, but such a defect has not been directly demonstrated. With peroral intestinal biopsies and immunofluorescence and intracellular localization of apolipoprotein B within jejunal epithelial cells of five normal subjects and have shown that its content increases markedly after fat feeding. In two patients with abetalipoproteinemia no apolipoprotein B was seen by immunofluorescence techniques in the jejunal mucosa in the fasting state or after a fatty meal. Intestinal synthesis of apolipoprotein B appears not to occur in abetalipoproteinemia.

It is known that after small bowel resection, mucosal hyperplasia and increased cell turnover occur in the remaining intestine, particularly the ileum. At the end of their life span, epithelial cells are extruded into the bowel lumen. Comparative estimates of this cell loss may be obtained by collecting the DNA from a perfused gut segment. Male Sprague-Dawley rats underwent resection of 50 cm of proximal or distal small intestine or sham operation. One and six months after surgery, 50 cm of the remaining proximal or distal remnant were perfused with saline in vivo and the perfusate DNA was assayed. The DNA recovered from the perfusate of the distal remnant was at least twice as much as that from sham control segments. This was associated with comparable increases in mucosal weight, DNA, and protein concentration per centimeter of distal remnant. No significant changes were found in perfusate DNA recovered from proximal remnants. This correlated with minimal, if any, changes in mucosal weight, DNA, or protein concentration per centimeter of these remnants. Increased desquamation of epithelial cells may reflect the hyperplasia of intestinal mucosa after bowel resection. Recovery of intraluminal DNA may prove useful as an index of intestinal adaptation.

Glucagonlike substances in extracts of intestinal mucosa were already described in 1948 by Sutherland and deDuve (1), who used a bioassay technique for the identification. After the development of the first glucagon radioimmunoassays, Unger and co-workers (2,3) confirmed that intestinal extracts contained peptides that "crossreacted" in the glucagon radioimmunoassay [hence gut "glucagonlike immunoreactivity" (GLI)]. In 1968, the same group discovered that the gut GLIs consisted of at least two peptides, GLI I and II (4), both of which differed immunochemically from pancreatic glucagon and, therefore, necessarily had different chemical structures (4,5). Developments during the last decade in the field of peptide chemistry, particularly improved purification and sequencing techniques, have greatly advanced our knowledge of gut peptides, including the enteroglucagons, and the chemical structure of several of the members of this heterogenous group of peptides is now known. Furthermore, progress in the field of nucleotide and gene technology has also spread to this area of research, and although many problems remain unresolved, the progress made has sufficiently important implications to justify a review of the most recent advances.

The regional and topographic distribution of substance P-containing nerve fibers in the human and feline intestinal wall was studied by immunocytochemistry and radioimmunoassay. The concentration of substance P was measured in the different layers of the duodenum, jejunum, ileum, and colon. In both humans and cat, substance P fibers were fairly numerous, and the substance P concentration was comparatively high in the smooth muscle layer, including the myenteric ganglia. In humans, but not in cat, substance P fibers were numerous, and the substance P concentration was also high in the mucosa. Substance P-containing nerve cell bodies were observed in the myenteric ganglia of both species. In the submucous ganglia, such nerve cell bodies were seen in the human intestine only, suggesting that they represent the origin of the numerous mucosal substance P fibers in this species. Previous studies have revealed a relative paucity of substance P fibers in the intestinal mucosa of several mammals, such as mouse, rat, and pig. The cat can now be added to those having few mucosal substance P fibers, whereas humans seem to be notably rich in such fibers, suggesting that substance P may play a role in the regulation of mucosal functions in the human intestine.

Extracts of human colon mucosa and muscle were fractionated by gel filtration and ion exchange chromatography and molecular forms of vasoactive intestinal peptide (VIP) estimated by radioimmunoassay. Total concentrations of immunoreactive VIP in mucosa were 237.1 +/- 53.9 and 119 +/- 26.0 pmoles per g in muscle. In muscle, over 90% of the immunoreactivity was accounted for by a single component indistinguishable from porcine VIP by gel filtration and ion exchange chromatography. In contrast, mucosa contained four components with VIP-like immunoreactivity. One of these had the chromatographic properties of porcine VIP; the others were less positively charged. Two of the new VIP-like components had gel filtration properties similar to those of VIP and are, therefore, probably of similar size to the porcine octacosapeptide, and the remaining component emerged later on Sephadex and is, therefore, probably a smaller peptide. Immunoreactive VIP in muscle extracts is believed to originate from nerve fibers, and this form of VIP is likely to be identical to the previously characterized form of VIP. However, immunoreactive VIP in mucosal extracts may originate either from endocrine cells or nerve fibers. The possibility arises, then, that there are different immunoreactive forms of VIP in nerves and endocrine cells.

Measurements of pH in the microclimate on gastric and duodenal mucosa were made during gastrointestinal endoscopy in 21 normal subjects and 9 duodenal ulcer patients. In controls luminal and juxtamucosal mean pH (+/- SEM) of 3.29 +/- 0.3 and 4.48 +/- 0.25 were recorded respectively in distal esophagus, 2.01 +/- 0.17 and 4.84 +/- 0.37 in gastric fundus, 1.82 +/- 0.12 and 5.5 +/- 0.15 in body, 3.52 +/- 0.34 and 5.42 +/- 0.29 in antrum, 6.89 +/- 0.21 and 7.16 +/- 0.13 in duodenal cap, and 6.84 +/- 0.19 and 7.03 +/- 0.19 in proximal duodenal loop. When the lumen of esophagus and duodenum were perfused with acid (pH 2) luminal and mucosal pH values were 2.18 +/- 0.11 and 4.08 +/- 0.41 in esophagus, 2.57 +/- 0.15 and 6.74 +/- 0.13 in duodenal bulb, and 2.44 +/- 0.14 and 6.39 +/- 0.2 in duodenal loop. Juxtamucosal pH in fundus, body, and duodenum remained near neutral when luminal pH was 1.5, but in distal esophagus and antrum it fell sharply at luminal pH values below 3. In duodenal ulcer patients juxtamucosal pH in the cap was significantly lower than that in normals at luminal pH values below 3. These studies support the hypothesis that a "mucus-bicarbonate" barrier inhibits mucosal acidification in humans and that duodenal mucosa in ulcer patients is less able to maintain a neutral zone adjacent to it in the face of luminal acid.

Immunoreactive secretin in hydrochloric acid extracts is relatively constant in the duodenum and proximal jejunum of pig and dog (3 microgram per g), but peaks in the distal duodenum of guinea pig (1 microgram per g), no secretin being detectable in the ileum of these species. Secretin is relatively constant throughout the small intestine of the rat and rabbit (0.4 to 0.1 microgram per g). Sephadex gel filtration patterns of all species and throughout the gastrointestinal mucosa revealed primarily a single peak with elution characteristics identical with that of purified or synthetic porcine secretin. The "big" secretin prominent in Boots secretin may be an alteration product perhaps attributable to chemicals used to stabilize the preparation. The "intermediate secretin" described by others has not been detected. It is concluded that in a variety of mammalian species most of the immunoreactive secretin extractable from the intestinal tract lies distal to the proximal duodenum and is not distinguishable from duodenal secretin in terms of molecular size.

In indirect immunofluorescence tests, antibodies against pure porcine cholecystokinin (CCK) have detected specific CCK cells in the duodenal and jejunal mucosa of the dog and man. The CCK cells were scattered in the epithelium of the crypts, although some were in the villi. No CCK cells were found in the stomach, pancreas, terminal ileum, or colon. Some pyloric G cells also showed some reactivity with CCK antiserum, but absorption of CCK antiserum with gastrin C terminal pentapeptide prevented the staining of pyloric cells and provided specific staining of intestinal CCK cells. Anti-human gastrin I serum stained some intestinal cells too. Most of such cells did not react when gastrin antiserum was absorbed with pure CCK (a treatment that did not prevent the staining of pyloric gastrin cells); they were interpreted as cross-reacting CCK cells rather than as intestinal gastrin cells.

The present report describes the morphologic findings in sequential colorectal biopsy specimens obtained from 2 patients who within 1.5-2.5 yr developed collagenous colitis. The initial biopsy specimens revealed an acute nonspecific inflammatory reaction in the colorectal mucosa. An intermediate stage, characterized by edema and slight fibrosis in the subepithelial region of the colorectal mucosa, preceded the final development of collagenous colitis. We suggest that the mechanism that leads to the formation of a thick, bandlike subepithelial collagenous deposit in the colorectal mucosa may be triggered off by some inflammatory or toxic stimulus.

The intramural distribution of the proenkephalin A-derived peptides Leu5-enkephalin, Met5-enkephalin, Met5-enkephalin-Arg6-Phe7, and Met5-enkephalin-Arg6-Gly7-Leu8 was studied throughout the human gastrointestinal tract. A parallel distribution was found of Leu5/Met5-enkephalin, measured with a Leu5-enkephalin antiserum that cross-reacts about 30% with Met5-enkephalin, and of Met5-enkephalin-Arg6-Phe7-immunoreactivity and Met5-enkephalin-Arg6-Gly7-Leu8-immunoreactivity. In each case, high tissue concentrations were present in the submucosa and muscularis corresponding to the pyloric sphincter. Taking all different regions together, a high correlation was revealed between tissue levels of Leu5/Met5-enkephalinlike peptides and Met5-enkephalin-Arg6-Gly7-Leu8-like peptides (r = 0.89), as well as between Met5-enkephalin-Arg6-Gly7-Leu8-like peptides and Met5-enkephalin-Arg6-Phe7-like peptides (r = 0.75). Met5-enkephalin-Arg6-Phe7 immunoreactivity was accounted for by a major peak (87% +/- 3% of total immunoreactivity) coeluting with the standard peptide in Sephadex G-50 chromatography and largely composed of the authentic heptapeptide, as shown by reverse-phase high-performance liquid chromatography. Leu5/Met5-enkephalin immunoreactivity was separated by high-performance liquid chromatography into peaks composed of Leu5-enkephalin and Met5-enkephalin. Allowing for Met5-enkephalin immunoreactivity in the assay used, the apparent Leu5/Met5-enkephalin molecular ratio was approximately 1:4. The high concentration of all peptides studied at the pyloric junction suggests a rich enkephalin-containing innervation at this level, in keeping with the proposed involvement of an enkephalinergic mechanism in the control of pyloric function.

Somatostatin and vasoactive intestinal peptide were measured by radioimmunoassay in acetic acid extracts of tissue samples from the digestive tracts of seven adult cats. The concentration of immunoreactive somatostatin in antral mucosa (2926 +/- 978 pmol/g wet weight, mean +/- SE) was far above the concentrations in the head of the pancreas (442 +/- 94), distal duodenum (377 +/- 91), and ileum (355 +/- 48). Large amounts of immunoreactive vasoactive intestinal peptide were recovered from both mucosal and muscular layers of gastrointestinal tract; the highest values were observed in the muscular layer of the cecum (579 +/- 82 pmol/g) and the mucosa of the right colon (564 +/- 244). The mucosal/muscular layer ratio of vasoactive intestinal peptide concentrations increased caudally from 0.20 in the esophagus to over 1 in the ileum and colon. Upon gel filtration on G-50 Sephadex, somatostatin in the antrum, duodenum, and pancreas were eluted as a predominant peak in the volume of the tetradecapeptide, but somatostatin from the ileum and cecum was associated with a major faster component. Vasoactive intestinal peptide in the muscular layer of gastrointestinal tract and in mucosa of antrum and duodenum consisted essentially of a single form, which coeluted with the purified porcine peptide; a slower component was detected in the mucosa of the ileum and cecum. These reults indicate molecular heterogeneity of both immunoreactive somatostatin amd vasoactive intestinal peptide in the digestive tract of cats, and suggest that the variable ratio of the different molecular forms of each peptide along the gastrointestinal tract may reflect regional specificity of biologic effects and metabolism.

Small intestinal absorptive function was investigated in genetically diabetic mouse model (C57 BL/KSJ dbm) in order to determine the long-term effects of genetic and uncontrolled diabetes mellitus on intestinal function. Initial rates of uptake of nonmetabolizable glucose analogs, beta-methyl-D-glucoside and 3-O-methyl-D-glucose were determined in diabetic mice and their littermate controls using everted sacs from proximal and distal halves of the intestine. In addition, intestinal weight, intestinal length, mucosal protein, and DNA were measured. There were no significant differences between controls and diabetics in rates of uptake by either proximal or distal segments. Kinetic characteristics of uptake, Km and Vmax, were similar in controls and diabetics. These results clearly demonstrate that intestinal transport of monosaccharides is not altered in genetic diabetes, and therefore are in contrast to augmented transport reported in the early phase of drug-induced diabetes but similar to the results observed in chronic drug-induced diabetes. However, diabetic mice exhibited stimulated intestinal growth similar to rats with chronic drug-induced diabetes.

Vasoactive intestinal peptide (VIP) levels were quantified by radioimmunoassay in three strata of hamster bowel wall, namely, epithelial cells isolated by a vibration technique, scrapings of the vibrated intestine, and in the remaining muscle. Expressed on a wet weight basis, highest levels of the peptide (as a percentage of total bowel wall content) were found in the muscle (72%) followed by high levels in scrapings of denuded villi (27%). Villus and crypt epithelial cells, isolated as one fraction, contained very low levels (less than 1%) of total VIP. To correct for differences in water content of the various strata, data were also calculated on the basis of tissue protein. When expressed in this manner, scrapings of denuded villi and the remaining muscle, both areas of dense autonomic innervation, were virtually equal in their VIP content (mean +/- SE, 9.5 +/- 4 and 11 +/- 2 ng per mg of protein, respectively). However, the VIP concentration of the cells was over two orders of magnitude less than the muscle or scrapings of denuded villi (0.02 +/- 0.007 ng per mg of protein). These results suggest that VIP is concentrated with or near the neural elements of the gut.

Insulin has been proposed as an important factor in the regulation of growth and differentiation of the small intestine. In the newborn miniature pig, we induced significant physiologic increases in serum insulin and the insulin/glucagon ratio without altering serum glucose, beta-hydroxybutyrate, glucagon, cortisol, T3, and T4 using glucose-based total parenteral nutrition (TPN) in one group (group G) compared with a combination of glucose and fat in another group (group G/F). Control animals were sham-operated and fed a pelleted diet (group OC). Duodenal villus surface area and mucosal height were significantly greater in group G/F compared with group G. No other differences between the TPN groups were found in small intestinal growth, mucosal protein, deoxyribonucleic acid and ribonucleic acid content, and disaccharidase activities. As anticipated, group OC demonstrated increased intestinal length, weight, and villous surface area compared with the TPN groups. Ileal sucrase and jejunal and ileal maltase activities were greater in the TPN groups compared with those in group OC. Physiologic changes in serum insulin and the insulin/glucagon ratio induced by the TPN fuel mix do not appear to have altered small intestinal growth, composition, and differentiation in the healthy small intestine.

The distribution of sigma and phencyclindine binding sites in the gastrointestinal tract of the guinea pig was studied by autoradiography after in vitro incubation of tissue slices with (+)3H-SKF 10,047 and 3H-1-1-[(2-thienyl)cyclohexyl] piperidine to locate sigma and phencyclidine receptors, respectively. A dense distribution of sigma binding sites was observed in the mucosa and in the submucosal plexus, particularly at the level of the fundus and duodenum. Muscular layers were only marginally labeled. No phencyclidine binding site could be demonstrated in any area. This selective distribution suggests a functional role of sigma receptors mainly in the control of endocrine or exocrine secretions, or both.

Luminal nutrients are a major effector of intestinal adaptation. Amino acids are trophic to the intestine, but their role in regulating amino acid transport is not well documented. The presence of several distinct amino acid transport systems raises the question of whether adaptation is class-specific. Studies were carried out in parenterally nourished rats receiving a 7-day jejunal infusion of a 3% solution of either aminoisobutyric acid, aspartic acid, glutamine, histidine, lysine, or valine. While all amino acids were trophic to the intestine, their effects on the in vitro uptake of 0.1, 1.0 and 10.0 mM aspartic acid, lysine, and valine (representative acid, basic, and neutral amino acids) were variable and nonspecific. Compared to controls receiving either total parenteral nutrition alone or total parenteral nutrition plus luminal saline, prior lysine and aspartic acid infusion significantly increased in vitro uptake of all three amino acids tested, whereas valine had little effect on transport. No effect on transport was seen with glutamine (actively metabolized by the intestine as is aspartic acid), aminoisobutyric acid (a nonmetabolizable amino acid congener), or histidine (the most trophic amino acid). In conclusion, while individual amino acids cause an adaptation of amino acid uptake, the effects are nonspecific and independent of their metabolic or trophic potential.

Class II histocompatibility antigens, known to be present on immunocompetent cells, were recently demonstrated on enterocytes. Because of their role in antigen presentation and immune response regulation, HLA-DR antigens were studied in patients with celiac disease. Cryostat sections of jejunal biopsy specimens were stained with several anti-DR monoclonal antibodies using an avidin-biotin peroxidase technique. Thirty patients with celiac disease either active (n = 5), in remission (n = 10), or in relapse (n = 15) were compared with 16 controls, 9 with a normal mucosa and 7 with a flat mucosa but without celiac disease. In celiac patients with active disease or in relapse, enterocytes were heavily stained on the surface epithelium and the crypts. This contrasted with the absence of crypt staining in the biopsy specimens of the other patients. Increase in DR expression was associated with an increase in the number of T8(+) lymphocytes in the crypts. Modulation of DR expression by enterocytes may be involved in the pathogenesis of celiac disease.

Pharmacologic characteristics of muscarinic receptors in the muscularis mucosae of the guinea pig esophagus were examined in vitro and compared with those of the longitudinal muscle of the guinea pig ileum. All cholinomimetics tested produced a sustained contraction of the muscularis mucosae, in a concentration-dependent manner, which was associated with a small biphasic change in membrane potential, initially a hyperpolarization followed by a depolarization. The contractile response was hardly modified by verapamil, but was depressed by calcium removal from the bathing medium. Both atropine and pirenzepine antagonized the contractile response in a competitive manner, with higher pA2 values than those in the ileum. These results indicate that the muscarinic receptors of the muscularis mucosae of the guinea pig esophagus mainly link with calcium ion channels which are independent of changes in membrane potential and that their subtype populations are probably pharmacologically distinct from those in the ileal longitudinal muscle.