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Biomedical subjects

I M Modlin

Publications and source records attributed to I M Modlin.

At least 109 records · Page 6Linked to original sources

Unique permeability barrier of the apical surface of parietal and chief cells in isolated perfused gastric glands.

Although many factors can influence intracellular pH (pHi), some of the most important are those that involve the movement of acids and bases across the cell membrane. We will discuss recent results concerning barriers to the movement of H+, NH3 and CO2 across the apical cell membranes of gastric gland cells. Cell membranes are generally highly permeable to small, lipophilic molecules such as NH3 and CO2. In fact, only two examples are known of membranes relatively impermeable to NH3 and none membranes permeable to CO2. We recently developed a technique for perfusing the lumen of a single hand-dissected gastric gland on the stage of a microscope, while monitoring pHi with a fluorescent dye. We observed the expected pHi changes when we exposed the basolateral (i.e. blood-side) membrane to a pH 6.4 solution (a large, rapid pHi decrease), to a pH 7.4 solution containing approximately 0.3 mmoll-1 NH3 (a large and rapid pHi increase) or to a pH7.4 solution equilibrated with 1% CO2 (a rapid pHi decrease of -0.08). However, pHi was not significantly affected by perfusing the lumen with a pH 1.4 solution, with a pH 7.4 solution containing as much as 2.7 mmoll-1 NH3 or with a pH 6.1 solution equilibrated with 100% CO2. These data indicate that a barrier at or near the apical membrane has a uniquely low permeability to H+, NH3 and CO2.

Ammonia↗

Recent advances in the localization and surgical management of duodenal gastrinomas.

Duodenal gastrinomas are now more frequently recognized as the source of hypergastrinemia in patients with Zollinger-Ellison syndrome. The cell lineage of duodenal gastrinomas may differ from that of pancreatic gastrinomas, which accounts for variations in their clinical behavior. Attempts to localize the submucosal tumors are difficult and are limited by their small size. Intraoperative endoscopic transillumination, selective intra-arterial secretin injection, and duodenotomy with mucosal eversion are currently the most sensitive and reliable methods of localization. Endoscopic ultrasonography and somatostatin scintigraphy further enhance the accuracy of preoperative localization of these tumors. Current information based on cure rates and survival data mandates a primary surgical approach in patients with either the sporadic or the multiple endocrine neoplasia type 1-associated form of the disease. Thus, wide local resection of duodenal gastrinomas with removal of all tumor-bearing lymphatic tissue and acid inhibitory pharmacotherapy (proton pump inhibition) may yield 5-year survival rates of 80% to 90%. Similarly, in patients with pancreatic and duodenal gastrinomas as a manifestation of multiple endocrine neoplasia type 1, the additional enucleation of pancreatic lesions with or without distal pancreatectomy has resulted in cure rates of 67% to 100%.

Diagnostic Imaging↗

Gastrin and colon cancer. Clarifying the controversy.

Controversy exists about the relationship between hypergastrinemia and colon cancer. Conflicting lines of evidence may be interpreted to support a variety of views regarding the link between the two. Although some experimental and clinical data suggest a strong correlation, other studies refute this. It is likely that the actual situation lies between these two viewpoints; the complex nature of the relationship between carcinogenesis and putative gut hormones makes a definitive answer unlikely. Nevertheless, a critical reading of the recent literature suggests that hypergastrinemia does not play a direct role in colorectal carcinogenesis. Certain subgroups of patients with elevated serum gastrin levels and tumors that possess gastrin receptors may have accelerated tumor growth. Further study of this issue is warranted to elucidate the role of the gastrointestinal hormonal milieu in colorectal neoplasia.

Colonic Neoplasms↗

Inhibition of acid secretion in gastric parietal cells by the Ca2+/calmodulin-dependent protein kinase II inhibitor KN-93.

A novel Ca2+/calmodulin-dependent protein kinase II (CaM Kinase II) inhibitor, KN-93 potently inhibits gastric acid secretion from parietal cells. As previously reported (1), treatment of parietal cells with a selective inhibitor of CaM kinase II, KN-62 resulted in the inhibition of cholinergic-stimulated rabbit parietal cell secretion, whereas it failed to inhibit the histamine and forskolin response. In contrast effects of carbachol, histamine and forskolin were significantly inhibited by KN-93 with an IC50 of 0.15, 0.3 and 1 microM, respectively; these effects occurred without any changes in intracellular cyclic AMP and Ca2+ levels. In the present study we investigated the mechanism by which KN-93 acts upon the acid-secreting machinery of gastric parietal cells. Neither redistribution of the proton pump activity nor the morphological transformation were affected by KN-93. The drug only weakly inhibited the H+, K(+)-ATPase activity but strongly dissipated the proton gradient formed in the gastric membrane vesicles and reduced the volume of luminal space. Thus KN-93 acts at pH gradient formation whereas KN-62 acts only at CaM Kinase II.

1-Methyl-3-isobutylxanthine↗

Identification of a small GTP-binding protein, Rab25, expressed in the gastrointestinal mucosa, kidney, and lung.

Small GTP-binding proteins have been implicated in the regulation of many dynamic cellular processes. The carboxyl termini of parietal cell small GTP-binding proteins were cloned using a 3'-rapid amplification of cDNA ends (RACE) technique with a degenerate oligonucleotide primer based on the WDTAGQE consensus GTP-binding sequence. Six out of 53 clones demonstrated a novel Rab sequence, now designated Rab25. The complete sequence was obtained using 3'-RACE and revealed a deduced amino acid sequence having 63% identity with Rab11. The deduced amino acid sequence demonstrated a carboxyl-terminal CCQNI and also a novel GTP-binding site sequence of WDTAGLE. Nevertheless, recombinant Rab25 was able to bind GTP on blot. A major 1.2 kilobase Rab25 message was detected throughout the gastrointestinal mucosa and in lung and kidney tissue. No message was detected in brain, heart, liver, or skeletal muscle. In gastric tissue, Rab25 was absent in the bowel wall; among mucosal cells, it was highly enriched in parietal cells compared to chief cells. Rab25 mRNA was also detected in several colon carcinoma lines including LIM1215 and HT-29. The results indicate that Rab25 represents a novel member of the Rab family with an epithelial distribution.

Amino Acid Sequence↗

Cyclic AMP-mediated release of peptide YY (PYY) from the isolated perfused rabbit distal colon.

We have investigated the role of the colonic nervous system in regulating the release of peptide YY (PYY) from the isolated perfused rabbit distal colon. In addition, we have studied the role of cAMP- and Ca(2+)-dependent mechanisms in mediating the release of PYY. We investigated three agents which stimulate increases in intracellular cAMP (vasoactive intestinal polypeptide (VIP), cholera toxin, and forskolin) and three agents which raise intracellular Ca2+ concentrations (substance P, carbachol, and the calcium ionophore A23187). The three cAMP-dependent agents, VIP (10(-7) M and 3 x 10(-7) M), cholera toxin (100 micrograms), and forskolin (10(-6) M and 10(-5) M) significantly stimulated release of PYY (P < 0.05). Tetrodotoxin (3 x 10(-6) M) did not alter forskolin (10(-5) M) stimulated release of PYY. Substance P (10(-9) M and 10(-8) M), carbachol (10(-5) M), and A23187 (10(-6) M) failed to stimulate the release of PYY. These results suggest that the colonic neurotransmitter VIP participates in the modulation of PYY release from rabbit distal colons. Further, these studies suggest that release of PYY is mediated by cAMP-dependent pathways.

Acetylcholine↗

Transforming growth factor-alpha (TGF alpha) inhibition of parietal cell secretion: structural requirements for activity.

Parietal cells of the gastric fundus produce transforming growth factor-alpha (TGF alpha) which functions as a potent inhibitor of acid secretion. We have previously demonstrated that TGF alpha can inhibit aminopyrine uptake in isolated rabbit parietal cells. In this study, we have evaluated the components of TGF alpha structure which determine its ability to inhibit parietal cell function. Both human and rat TGF alpha inhibited histamine stimulation by increasing the EC50 for agonist stimulation. Three fragments containing the third loop domain of TGF alpha (rat TGF alpha 34-43, human TGF alpha 34-43 and human TGF alpha 34-50) all inhibited histamine stimulation with IC50 values 20, 33 and 4-fold higher, respectively, than that of the native molecule. Rat TGF alpha inhibited carbachol stimulation throughout an agonist dose response. Human TGF alpha was only effective in inhibiting carbachol if incubations were performed in the presence of air rather than 100% O2. In air incubation, all three of the TGF alpha fragments inhibited carbachol stimulation but, in contrast to the effects on histamine, the peptides all were virtually equipotent with the native molecule. The human sequence fragments, like the native human TGF alpha, elicited no inhibition when incubations were performed in the presence of 100% O2. The results suggest that there are pharmacological differences in the response of isolated parietal cells to TGF alpha-mediated inhibition of histamine and carbachol. In addition, in contrast with previous investigations on the mitogenic action of TGF alpha, third loop fragments of TGF alpha retain the capacity to inhibit aminopyrine accumulation.

Amino Acid Sequence↗

Counterregulation of a prokinetic calcium-dependent mechanism by cAMP-dependent agents in isolated segments of terminal ileum.

The ill-understood complex of the irritable bowel syndrome comprises a group of intestinal motility disorders characterized by increased intraluminal pressures and decreased transit times. Elucidation of mechanisms which modulate gut motility may lead to the development of rational therapy for this prevalent problem. The purpose of this study was firstly to evaluate the interaction of cAMP-dependent agents (vasoactive intestinal polypeptide (VIP), norepinephrine (NE), and forskolin (FK)) on carbachol (Ca2+)-initiated motility and secondly to determine if a neural component of motility modulation existed by testing if the effect of cAMP-dependent agents was reversed by tetrodotoxin-induced neural blockade. Motility was measured in isolated segments of terminal ileum harvested from rabbits using perfusion manometry and quantitated by integration, expressed as mm Hg/min. Carbachol caused a concentration-dependent increase in measured motor activity (half-effective dose = 10(-7) M). VIP, NE, and FK each caused a concentration-dependent inhibition of carbachol-stimulated phasic contractions. TTX 10(-6) M failed to block the inhibitory actions of NE. In conclusion, these results suggest that cAMP-dependent mechanisms may inhibit gut motility induced by a cholinergic (Ca2+)-mediated agonist and that this process is mediated by a nonneural mechanism.

Animals↗

Biology and management of the midgut carcinoid.

Midgut carcinoid tumors derive from gut entoderm. These tumors may cause a complex of symptoms comprising the carcinoid syndrome by secreting a wide variety of bioactive agents in addition to serotonin. Such symptoms generally follow metastases to the liver but may also occur in primary ovarian or retroperitoneal tumors. After localization and biochemical characterization, the bioactivity of these tumors should be blocked by octreotide, sometimes in combination with other pharmacologic antagonists, so that primary resection may be performed safely. If curative resection is impossible, then a cytoreductive management scheme should be employed that includes surgical debulking and hepatic arterial embolization, followed by palliation with octreotide.

Algorithms↗

Influence of chloroquine on diet-induced pancreatitis.

Recent investigations have suggested that digestive zymogens may become activated within the acinar cell during acute pancreatitis. While the molecular events responsible for intracellular zymogen activation remain unknown, several potential enzymatic pathways require an acidic pH to optimally proceed. We therefore proposed that manipulation of subcellular pH might alter the course of experimental pancreatitis. Chloroquine, a weak base that raises the pH of acidic subcellular compartments, was administered to young female mice in which pancreatitis was induced by a choline-deficient, ethionine-supplemented (CDE) diet. Control animals were maintained on regular laboratory chow. Examination of isolated pancreatic acini using acridine orange cytofluorescence demonstrated expansion of acidic subcellular compartments in animals fed the CDE diet. These compartments were effectively neutralized in animals receiving chloroquine. Animals receiving continuous infusions of high-dose chloroquine demonstrated a significant (p < 0.05) decrease in free pancreatic tryptic activity as well as improved survival. These changes were also associated with decreased trypsinogen content in animals treated with high-dose chloroquine, suggesting an additional potential effect of chloroquine on zymogen synthesis and accumulation. One explanation of these findings is that a low-pH compartment may be important in the pathogenesis of diet-induced pancreatitis.

Acute Disease↗

Bile salt inhibition of motility in the isolated perfused rabbit terminal ileum.

The effects of bile on small bowel motility were studied in isolated, perfused rabbit terminal ileum. It was proposed that bile delivery into the distal ileum would inhibit ileal motor activity, by peptide YY (PYY) release and therefore the effect of luminal bile on motor activity was examined and PYY release measured. Luminal bile and taurocheodeoxycholic acid (10 mmol) inhibited ileal motor activity. Arterial infusion of venous effluents from a bile inhibited ileum suppressed motor activity in a second isolated ileum. This shows the presence of a humoral inhibitor of ileal motor activity. Luminal bile increased venous PYY concentrations (42.5 (8.5) to 502 (46.2) pmol/l; p < 0.01) and increased bile salt values (1.7 (0.36) to 88.6 (5.6) 10 mumol/l/l; p < 0.005). Arterial infusion of taurocheodeoxycholic acid at concentrations found in the venous effluent (100 mumol/l/l) suppressed motility (p < 0.001) but infusion of PYY at concentrations in the venous effluent (500.0 pmol/l) failed to inhibit motility. Furthermore, PYY antagonist, PYX 1, failed to reverse the bile induced inhibition of motility. Luminal bile salts inhibit terminal ileal motility and this is independent of PYY release. By slowing motility, bile salts may participate in their own absorption by the 'ileal pump' and in the 'ileal brake' mechanism.

Animals↗

Deoxycholate is an important releaser of peptide YY and enteroglucagon from the human colon.

Peptide YY (PYY) and enteroglucagon are hormonal peptides found in endocrine cells of the distal intestinal mucosa. Although it is known that plasma concentrations of both peptides increase in response to feeding, the mechanism by which ingested food causes release of colonic hormones is not understood. The release of PYY and enteroglucagon was measured in response to intraluminal stimuli in 176 patients having investigative colonoscopy. Introduction of air, saline (isotonic and hypertonic), glucose (isotonic and hypertonic), oleic acid (without bile salts), and casein hydrolysate all failed to release PYY but glucose caused a small but significant increase in enteroglucagon concentrations. In contrast with the lack of effect of nutrients, infusion of deoxycholic acid produced a rapid and marked dose responsive increase in plasma PYY concentrations when introduced into the sigmoid colon. PYY release was statistically significant at doses between 3.3 mM to 30 mM; for example 10 mM deoxycholate caused a sixfold increase in plasma PYY concentrations. Infusion of 10 mM deoxycholate into the transverse colon or caecum produced an increase of PYY that was similar to the responses in the sigmoid colon. There was also a significant release of enteroglucagon in response to infusion of this bile salt into the sigmoid colon at doses between 3.3 mM and 30 mM. The enteroglucagon response to 10 mM deoxycholate was similar in all three colonic regions. When oleic acid was added to deoxycholate as an emulsion, the release of PYY and enteroglucagon was similar to that seen with the bile salt alone. These findings suggest that bile salts may play an important part in the control of colonic endocrine function and may explain the increased circulating concentrations of colonic regulatory peptides that are seen in malabsorption states and after small bowel resection in humans.

Aged↗

Tyrosine kinase activities in the modulation of stimulated parietal cell acid secretion.

The effects of protein tyrosine kinase (PTK) activities on the modulation of secretion were investigated in isolated rabbit parietal cells. Two classes of inhibitors, genistein (an inhibitor of both soluble and membrane-associated PTK activities) and an erbstatin analogue (an inhibitor of membrane-associated PTK activities), were tested against both secretagogue stimulation as well as transforming growth factor-alpha (TGF-alpha) inhibition. Pretreatment of rabbit parietal cells with 10(-7) M rat TGF-alpha resulted in inhibition of both histamine- and carbachol-stimulated [14C]-aminopyrine (AP) accumulation. TGF-alpha inhibition was totally reversed by simultaneous pretreatment of cells with 50 microM genistein or an erbstatin analogue, indicating that a receptor-associated PTK activity is likely involved in the inhibition of parietal cell secretion. Furthermore, genistein, but not the erbstatin analogue, potentiated histamine-stimulated AP accumulation with a change in EC50 from 1.9 to 0.5 microM. Similarly, genistein, but not the erbstatin analogue, potentiated the response to forskolin with a change in EC50 from 1.5 to 0.1 microM. Genistein had no effect on stimulation of AP uptake by either dibutyryladenosine 3',5'-cyclic monophosphate or carbachol. In addition, genistein failed to increase histamine or forskolin stimulation beyond the maximal level and had no significant effect on either cellular adenosine 3',5'-cyclic monophosphate production or intracellular Ca2+ concentration. These results suggest that a PTK-protein phosphotyrosine phosphatase system may be involved in the potentiation of the histamine signal by a mechanism independent of adenylate cyclase activation.

Animals↗