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PubMed1990

Failure of 1,25-(OH)2-vitamin D3 to affect calcium flux across sheep rumen wall mucosa in vitro.

Isolated pieces of mucosa from the sheep rumen wall mounted between half-chambers were used in conventional incubation studies (1 h, 39 degrees C) with buffer solutions resembling rumen fluid (= mucosal fluid) and extracellular fluid (= serosal fluid), respectively. The mucosal-to-serosal (ms) flux of calcium was studied by use of 45CaCl2 with and without 1,25-(OH)2-vitamin D3 in the serosal incubation fluid. Different concentrations of the vitamin had no effect on Ca ms-flux.

Animals↗
PubMed1985

Transport of sodium fluorescein between the mucosal and muscular layers in man, possibly indicating functional serially-coupled exchange vessels.

In sixteen patients subjected to intestinal surgery, the transport of sodium fluorescein (Na-F) was measured between the mucosal and serosal-muscular layers. Experiments were carried out in intestinal anastomosis by fluorescein flowmetry (FF). The blood-flow index of the mucosal layer was about twice the serosal-muscularis, per unit tissue volume. The temporal changes in fluorescence pattern from the two layers showed that more than 90% of Na-F, eliminated from the mucosal layer, was transported to the serosal-muscular layers. In six patients, in whom Na-F was instilled intraluminarily, fluorescence was seen on the outside of the intestine after 3 min. However, when the circulation was stopped before Na-F was instilled, no fluorescence was seen, this suggests that the transport was not only due to diffusion but also to convection. The results suggest that functional serially-coupled exchange vessels may exist within the intestinal wall, and that Na-F is transported both by convection and diffusion.

Biological Transport↗
PubMed1974

The influence of anaesthetics on the increase in the water permeability of the toad bladder induced by vasopressin.

1 Single lobes of the bladder of Bufo marinus were isolated and filled with, and suspended in, oxygenated Ringer solution. The fluid in contact with the outside (serosa) of the lobes had a total osmolarity of 225 m-osmol/litre, and that bathing the inside (mucosa) of 45 m-osmol/litre.2 Osmotic water flow from mucosa to serosa was measured by weighing the lobes every 30 minutes. It was negligible unless vasopressin was added to the serosal bath. Standard concentrations of either 1.25 or 6.25 mu/ml were used to render the bladder lobes permeable to water.3 The presence in the serosal medium of pentobarbitone or thiopentone in concentrations ranging from 0.25 to 2.5 mM, or of chloralose in concentrations ranging from 0.65 to 6.5 mM, diminished the increase in water permeability induced by vasopressin.4 The three anaesthetics exerted similar inhibitory effects on the action of vasopressin from the serosal and from the mucosal surface of the bladder.5 In the presence of a constant high concentration of anaesthetic, increasing the concentration of vasopressin over three orders of magnitude led to stepwise increases of osmotic water flow out of the lobes, although at every dose level the effect of vasopressin was depressed by the anaesthetic. However, it was not completely abolished even if the concentration of vasopressin was close to threshold.6 The increase in water permeability of the bladder induced by 3',5'-adenosine monophosphate (cyclic AMP) was also depressed by the three anaesthetics.7 Possible explanations of the findings are discussed.

Adenosine Monophosphate↗
PubMed1999

Gastric carcinoma confined to the muscularis propria: how can we detect, evaluate, and cure intermediate-stage carcinoma of the stomach?

OBJECTIVE: The most important surgical strategy for advanced gastric cancer is its detection at the curative stage. The aim of this study was to characterize the curable intermediate-stage gastric carcinomas. METHODS: Of 1120 consecutive patients who underwent gastric resection for primary gastric cancer from 1979 through 1996, 94 patients were histologically diagnosed as having cancer confined to the muscularis propria (mp cancer), analyzed clinicopathologically, and compared with patients with early and serosal cancers. RESULTS: The operative incidence of mp cancer was around 8% among cases of gastrectomy, and the ratio of mp cancer to advanced cancer began to increase in 1991. Mp cancer was at a statistically intermediate stage, between early and serosal cancers in terms of symptoms, surgical curability (96%), size and histology of the tumor, and the rate of lymph node metastasis (46%). Preoperative assessments of tumor depth were unclear using radiology and endoscopy; however, 35% of 31 cases studied were diagnosed precisely by endoscopic ultrasonography (EUS). Accuracy of lymph node metastasis diagnosis was the same (65%) by preoperative EUS and by surgeon; however, sensitivity of the surgeon's assessment was higher (69% vs 38%) and specificity of EUS was higher (83% vs 39%). The 5-yr survival rate was 85%, which was significantly better than that of serosal cancer and similar to that of early cancer. Patient outcome was not affected by lymph node metastasis or macroscopic type of tumor. CONCLUSIONS: Mp cancer should be considered an intermediate-stage cancer. Surgery with level 2 lymph node dissection should provide a cure rate similar to that for early cancer.

Aged↗
PubMed1993

Atrial natriuretic factor (ANF) does not affect ion transport in human intestine but does in porcine intestine.

The aim of this study was to test whether atrial natriuretic factor (ANF) exerts any effect on human intestinal ion transport, and the porcine intestine was used as a positive control of ANF's effects. Tissues from human proximal (n = 6) and distal (n = 6) colons, and from distal ileum (n = 6) were mounted in Ussing chambers, and short circuit current (Isc) was measured subsequent to serosal application of ANF (10(-6) M), 8-Br-cyclic guanosine monophosphate (8-Br-cGMP) (10(-4) M), and theophylline (10(-2) M). ANF did not affect Isc whereas 8-Br-cGMP increased Isc by 28 (8-53), 16 (3-36), and 16 (5-41) microA cm-2 in the distal colon (DC), proximal colon (PC) and distal ileum (DI), respectively. Likewise, transepithelial potential difference (PD) became more negative by 5.0 (0.6-8.9), 2.5 (0.4-4.0) and 0.9 (0.3-2.3) mV in DC, PC, and DI, respectively, subsequent to addition of 8-Br-cGMP. Isc and PD were further increased by theophylline. Additional radio-isotope flux studies in human colon revealed that ANF did not affect electroneutral sodium and chloride transport either. For comparison, ANF (10(-6) M) was administered to large intestinal tissues from young pigs in which ANF induced a significant increase in Isc which was comparable to the 8-Br-cGMP response in humans. The porcine Isc response was partly inhibited by chloride-free solution on the serosal side, by serosal application of bumetanide (10(-4) M) and BaCl2 (10(-3) M), and mucosal application of the chloride-channel blocker diphenylamine-2-carboxylate (DPC) (10(-3) M). Mucosal amiloride (10(-5) M) pre-treatment reduced baseline Isc but did not affect the porcine intestinal Isc response to ANF. In vitro radio-autography demonstrated specific binding sites for ANF in porcine distal colon, whereas no apparent labelling was observed in human distal colon. These findings suggest that the lack of effect of ANF on sodium and chloride transport in human distal ileum and colon is probably due to lack of ANF receptors. In the porcine intestine, however, the Isc response induced by ANF seems to involve stimulation of electrogenic chloride secretion, whereas electrogenic sodium absorption seems unaffected.

Adolescent↗
PubMed1995

Prostaglandin E2 release from dermis regulates sodium permeability of frog skin epithelium.

In the present study we have compared the effects of increased intracellular Ca2+ in whole frog skin and isolated epithelium (Rana temporaria). Cellular Ca2+ was increased by the use of the endoplasmic reticulum Ca2+ ATPase inhibitor, thapsigargin. Serosal addition of thapsigargin to the whole frog skin increased the Na+ transport by increasing the apical Na+ permeability. This could be blocked by the addition of indomethacin or by removal of Ca2+ from the serosal solution. The increase in Na+ transport was accompanied by an increased prostaglandin E2 release. This indicated that the response in Na+ transport was due to a Ca2+ dependent activation of the prostaglandin E2 synthesis. Addition of thapsigargin to isolated epithelia inhibited the Na+ transport and had no effect on the prostaglandin E2 release, though the prostaglandin E2 release from the isolated epithelia could be increased by the addition of arachidonic acid. Addition of prostaglandin E2 increased the cAMP contents of the isolated epithelia significantly, whereas thapsigargin had no significant effect on the cAMP level. Our results demonstrate that serosal addition of thapsigargin causes a release of prostaglandin E2 from the dermis below the transporting epithelium. The prostaglandin E2 diffuses to the epithelium where it activates the Na+ transport by increasing cellular cAMP. The epithelium itself does not contribute significantly to the prostaglandin E2 synthesis. Furthermore an increase in intracellular Ca2+ in the epithelial cells without a concomitant increase in prostaglandin E2 release leads to an inhibition of the active Na(+)-transport.

Amiloride↗
PubMed1989

Induction of peritoneal adhesions with small intestinal ischaemia and distention in the foal.

Twenty-two foals were divided into groups of intestinal distension and intestinal ischaemia as methods to induce peritoneal adhesions. In the first group, the lumen of a segment of distal small intestine was occluded without extramural vascular compromise and distended with lactated Ringer's solution to a constant pressure of 25 cm H2O for 2 h within the abdomen. The ischaemic group underwent 70 mins total vascular occlusion of identical segments of bowel. Serosal biopsies were obtained before and after each experimental procedure and following 60 mins of reperfusion. Similar biopsies were harvested from a control group of foals with no bowel occlusions. The foals were destroyed 10 days after surgery and tissues collected for histological and ultrastructural evaluation. Experimental and control mesothelial surfaces were denuded histologically immediately after experimental occlusions. Serosal oedema and cellular infiltration were observed following reperfusion of the ischaemic segments but were present immediately after 2 h of distension. All foals had developed bowel-to-bowel and bowel-to-mesentery adhesions of the experimental segments. Control foals under 30 days old exhibited mesenteric contraction and thickening of the isolated segment whereas those older than 30 days had little or no mesenteric thickening or contraction. Histologically, in the experimental segments, fibrous tissue had formed on the outer boundary of the original serosa, and new mesothelial-like cells were present on the surface of fibrous tissue in some areas. Some serosal fibrosis was also seen in most of the control segments.

Animals↗
PubMed1994

Studies on the transport in vitro of lysine, histidine, arginine and ammonia across the mucosa of the equine colon.

Discs of stripped mucosa from the proximal ventral colon were prepared immediately after slaughter of 8 Shetland cross-breed ponies. The mucosae were fixed in incubation chambers and used in incubation experiments to study the transmucosal fluxes of the amino acids lysine, histidine and arginine (150 min) and of ammonia (90 min). When the amino acid concentrations in the mucosal solution were in the physiological range (2.8-3.0 mmol/l) no transport to the serosal side of the tissue was found. When the concentrations were raised 10-fold, less than 2% of the mucosal amino acid pool was recovered in the serosal solution. Ammonia was transported across the mucosa at significant rates although only 63% of the ammonia that disappeared from the mucosal solution was found in the serosal solution. The findings show that the equine proximal colon is virtually impermeable to luminal free amino acids whereas ammonia is transported at rates equal to, or higher than those observed with rumen mucosa from sheep.

Amino Acids, Essential↗
PubMed1988

Involvement of prostaglandins in histamine-induced fluid and electrolyte secretion by rat colon.

Histamine increased the transmural potential difference across rat colon in-vivo and induced a net secretion of fluid. Both effects were inhibited by indomethacin. Histamine increased the potential difference and short-circuit current, and reduced tissue resistance in colonic sheets in-vitro. This response was reduced in the absence of chloride in the bathing medium or in the presence of serosal frusemide, suggesting that histamine stimulated electrogenic chloride secretion by the colon. The rise in short-circuit current induced by histamine was calcium-dependent since it was reduced in the absence of serosal calcium or in the presence of serosal verapamil. Indomethacin, a cyclo-oxygenase inhibitor, and mepacrine, a phospholipase inhibitor, both caused a dose-dependent inhibition of the electrical response of colonic sheets to histamine, without affecting the rise in short-circuit current induced by prostaglandin E2. The stimulation of chloride secretion induced by histamine in rat colon therefore appears to be mediated by an increased production of prostaglandins.

Animals↗
PubMed1989

Effect of auranofin on ion transport by rat small intestine.

Auranofin, applied either mucosally or serosally, increased the potential difference and short-circuit current generated by stripped sheets of rat mid-intestine in a concentration-dependent manner. In-vivo auranofin induced a net fluid secretion, suggesting that the rise in electrical activity represented a stimulation of anion secretion. Removal of chloride or addition of frusemide inhibited the auranofin-induced increase in short-circuit current, indicating that chloride was the anion involved and in the case of serosal auranofin this was confirmed by direct measurement of ion fluxes. The effects of both mucosal and serosal auranofin were calcium-dependent. The gold component of the auranofin molecule is probably responsible for its secretory actions as these were mimicked by chlorauric acid. The ability of auranofin to stimulate intestinal secretion may contribute to its diarrhoeagenic action.

Animals↗
PubMed1991

Reconsideration of inhibitory effect of metformin on intestinal glucose absorption.

New evidence that metformin increases intestinal glucose metabolism has necessitated a re-examination of the effect of metformin on intestinal glucose absorption. Normal 18 h fasted mice received an intragastric bolus of metformin 2 h before preparation of everted gut sacs from the proximal, middle and distal regions of the jejunum and ileum. Net mucosal glucose transfer from the intestinal lumen into the tissue was reduced by 15 and 28% after 50 and 250 mg kg-1 metformin, respectively (ANOVA, P less than 0.05). Net glucose transfer into the serosal fluid was reduced by 12 and 70% after 50 and 250 mg kg-1 metformin respectively (ANOVA, P less than 0.05 and P less than 0.01). The inhibitory effect of metformin on both the mucosal and serosal glucose transfer mechanisms was greatest in the middle portion of the small intestine. The results suggest that metformin decreases intestinal glucose absorption in a dose-dependent manner by effects on mucosal and serosal glucose transfer.

Administration, Oral↗
PubMed1995

The influence of uptake from the gastrointestinal tract and first-pass effect on oral bioavailability of (Z)-alkyloxyimino penicillins.

We have investigated the contribution of uptake from the gastrointestinal tract and first-pass effect to the poor oral bioavailability of a series of (Z)-alkyloxyimino penicillins in mice. Investigative studies in gut sacs and perfused small intestine demonstrated that these penicillins were able to pass across the mucosal epithelium although to a lesser extent than amoxycillin and cyclacillin, both of which exhibit excellent oral bioavailability in man and animals. In the jejunal gut sacs the mucosal to serosal flux for BRL 44154 was approximately half that of amoxycillin and four times less than that of cyclacillin, and for all, uptake was pH dependent. The serosal to mucosal fluxes were however similar for these compounds and significantly lower than mucosal to serosal fluxes, suggesting involvement of carrier mechanisms in uptake from the mucosal surface. The order of results for the alkyloxyimino penicillins paralleled that observed for oral bioavailability in the mouse. For the alkyloxyimino penicillins, between 5.5 and 9.9% was taken up from the perfused intestine, values which were significantly less than those for amoxycillin (13.2%) and cyclacillin (33.3%). However, uptake was concentration-dependent for BRL 44154 as it was for amoxycillin, thus confirming the possible use of carrier mechanisms in absorption. These observations suggest that the poor peripheral blood concentrations of the alkyloxyimino penicillins achieved after oral dosing were not a consequence of the inability of the compounds to cross the mucosal epithelium. The biliary clearance of the alkyloxyimino penicillins was, however, considerably greater than for amoxycillin and cyclacillin, a finding which may well have been a contributory factor to the comparatively low peripheral concentrations of BRL 44154 and its analogues achieved after oral administration.

Administration, Oral↗
PubMed1997

Intestinal absorption of stable cyclic glycylphenylalanine: comparison with the linear form.

The absorption, especially the stability and transportability, of the cyclic peptide cyclic glycylphenylalanine (cyclo(Gly-Phe)) and the linear peptides glycylphenylalanine, glycyl-D-phenylalanine and phenylalanylglycine have been studied in rat small intestine. Linear peptides were degraded on the mucosal side and only glycyl-D-phenylalanine appeared on the serosal side. However, cyclo(Gly-Phe) was stable on the mucosal side and appeared on the serosal side. Furthermore, the absorption clearance of cyclo(Gly-Phe) was higher than that of glycyl-D-phenylalanine. In the presence of the peptidase inhibitor bestatin, the degradation of linear peptides was reduced and linear peptides appeared on the serosal side, but only phenylalanylglycine, which is transported by the oligopeptide transporter, was absorbed faster than cyclo(Gly-Phe). The absorption clearance of cyclo(Gly-Phe) was reduced as its concentration was increased from 125 microM to 500 microM. Furthermore, the absorption clearance of cyclo(Gly-Phe) at 125 microM was reduced at 4 degrees C or in the presence of glycylsarcosine and cephalexin, which are transported by the oligopeptide transporter. These results indicated that cyclo(Gly-Phe) was stable enough to be absorbed and was transported in part by the oligopeptide transporter rather than completely by passive diffusion.

Animals↗
PubMed1983

Use of high performance liquid chromatography to study absorption and metabolism of purines by rat jejunum in vitro.

A simple and specific analytical method incorporating high performance (anion-exchange) liquid chromatography has been used to study the absorption and metabolism of some purine derivatives in an in vitro preparation of adult rat jejunum. When present in the lumen the purine bases guanine (4 X 10(-5)M), hypoxanthine or xanthine (10(-4)-3 X 10(-4)M) do not appear in the serosal secretions but the uric acid (UA) content of these secretions is increased. With UA in the intestinal lumen (10(-4)-3 X 10(-4)M) the serosal UA is increased, in some cases to a higher concentration than that in the lumen. With adenine in the lumen (10(-4)-10(-3)M) there is an increased appearance of UA and adenine also appears in the serosal secretions, but the concentration of adenine never exceeds that in the lumen. It is concluded that purines absorbed from the lumen are significantly metabolized to UA during translocation across rat jejunum. Negligible amounts of metabolites are found in the luminal solutions except for guanine which appears to be degraded in the lumen.

Absorption↗
PubMed1991

Cumulative dose-response curves for bethanechol-induced electrogenic secretion in rat jejunum in vitro: is tachyphylaxis a significant factor?

Electrogenic secretion was monitored as the short-circuit current (Isc) by an automatic voltage clamp across the isolated rat jejunum incubated in vitro. Responses to consecutive additions of bethanechol (1 mM) to the serosal surface showed tachyphylaxis. Addition of prostaglandin E2, however, after a single dose of 1 mM-bethanechol, induced an undiminished Isc response indicating that the tachyphylaxis was homologous. Dose-response curves for serosal applications of bethanechol obtained by serial-cumulative addition did not show tachyphylaxis when compared with those from a non-cumulative technique. The method of automatically recording the intestinal Isc with serial-cumulative addition of a serosal secretagogue gives dose-response curves free from tachyphylaxis.

Animals↗
PubMed1995

Epidermal growth factor stimulates the restitution of rat gastric mucosa in vitro.

Epidermal growth factor (EGF) is mitogenic to gastric mucosal cells, protects against gastric mucosal injury induced by ulcerogenic agents, and stimulates cell migration in many systems. Restitution, the rapid re-establishment of mucosal integrity following damage, involves epithelial cell migration and can be monitored by measuring the transmucosal potential difference (PD) of tissue mounted in an Ussing chamber. The role of EGF in restitution is poorly characterized. To assess the effect of exogenous EGF on restitution, samples of rat gastric mucosa were mounted in an Ussing chamber with a solution of 10 nM EGF bathing either the mucosal or serosal surface. Matched control tissues from the same animals were untreated. Mucosal damage was induced by exposure of tissues to hyperosmolar sodium chloride. Restitution following injury was monitored by measuring the recovery of transmucosal PD, which reflects the degree of mucosal integrity. Undamaged mucosa maintained a PD of approximately -30 mV for several hours (mucosa negative with respect to serosa). Mucosal exposure to 1.2 M NaCl for 2 min reduced the PD to near 0 mV. Thereafter, the PD returned in approximately 60 min to plateau at a value representing maximal recovery. Tissues mucosally treated with 10 nM EGF recovered following damage to a maximum of 60.2 +/- 5.1% of pre-exposure PD, while matched controls recovered to only 46.2 +/- 5.8%, a significant difference. Tissues serosally treated with EGF recovered to 61.0 +/- 3.2% and matched controls recovered to 51.1 +/- 2.9%, also a significant difference. These results suggest that both serosally and mucosally applied EGF stimulates the restitution of rat gastric mucosa in vitro.

Animals↗
PubMed1966

Amino acid transport in the goldfish intestine.

1. The serosal transfer of the following eight amino acids: threonine, alanine, serine, histidine, valine, methionine, phenylalanine and leucine, was measured using everted sacs of anterior intestine taken from goldfish acclimatized to 8 degrees C and incubated at 25 degrees C.2. All eight amino acids were actively transported and the serosal transfer correlated with the steady potential (P < 0.001) and with the amino acid-evoked potential (P < 0.05) measured on the same preparations.3. The goldfish rectum actively transported alanine and the steady potential was raised when alanine bathed the mucosa of the everted preparation.4. L-aspartic acid was partly transaminated to alanine by the goldfish anterior intestine; the rectum transaminated alanine to an unidentified amino acid which might have been serine, asparagine or glutamine or some mixture of these three.5. It is suggested that L-amino acids increase the ease by which sodium enters the mucosal cell but that it is the rate at which this sodium is transported across the basal membrane which determines the net serosal transfer of amino acids.

Alanine↗
PubMed1967

The in vitro transfer of bovine immune lactoglobulin across the intestine of new-born pigs.

1. Everted sacs of pig intestine, used soon after birth, maintained transmural potentials and transferred water and glucose to the serosal surface.2. Immune globulin, fed as bovine colostrum to the new-born pig, appeared in the serosal fluid of everted sacs during incubation in bicarbonate saline. The particular segment showing maximum transferring ability varied between limits and appeared to depend on the amount or concentration of colostrum fed to the pig. Sacs from unfed pigs incubated in bovine colostrum also transferred colostral IgG to the serosal fluid. This transfer was dependent on the concentration of colostral IgG in the incubation medium and became more pronounced in the middle third of the small intestine.3. Human serum albumin inhibited the transfer of colostral IgG and about twenty molecules of albumin were transferred for every molecule of colostral IgG, when both were presented together in equal concentration on the basis of weight, to the middle segment of the small intestine.4. Some of the immune globulin collected in vitro after feeding bovine colostrum was found in a degraded form, but the amounts present could not be estimated. There was no apparent degradation of immune globulin in the purely in vitro experiments.5. The in vitro transfer of bovine colostral IgG showed selectivity between molecules of albumin and colostral IgG, the nature of which warrants further study.

Animals↗