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PubMed1996

Differential effect of growth factors on intestinal wall components.

Both the mucosal and muscle layers respond to surgical manipulation of the small intestine. Various growth factors influence the mucosal response but less is known about changes in the muscle layers. Our aim was to evaluate the effect of epidermal growth factor (EGF) and octreotide (OCT) on changes in the intestinal wall induced by serosal patching. Twenty-four New Zealand white rabbits (3-4 kg) were studied. Group I (n=6) were unoperated controls. Group II (n=6) underwent creation of a 2 X 5-cm serosal patch in the ileum. Group III (n=6) and Group IV (n=6) received EGF (1.5 micrometer/kg/hr SQ) or OCT (1000 microgram/d SQ) after serosal patching. Wall composition, collagen content, and mucosal proliferation and enzyme activity of normal adjacent ileum were studied after 7 days. Serosal patching resulted in thickening of the mucosa and muscle layers (circular muscle: 189 +/- 54 vs 86 +/- 33 microns longitudinal muscle: 105 +/- 17 vs 55 +/- 12 microns P < 0.05) with increased proliferation and collagen content. EGF augmented the mucosal thickening (705 +/- 66 vs 573 +/- 94 microns, P < 0.05) and stimulated mucosal proliferation but inhibited the changes in muscle and collagen content. OCT inhibited the mucosal thickening and proliferation, but had less affect on the muscle. Disaccharidase activity was similar in all of the patched groups. We conclude the following. (1) Growth factors have differential effects on the intestinal wall components. (2) EGF stimulates the mucosa but has inhibitory effects on induced muscle changes. (3) Octreotide inhibits mucosal proliferation with less effect on muscle. (4) These observations have important implications for the therapeutic applications of these agents.

Animals↗
PubMed2002

Spatially selective laser coagulation of the gastric wall: a new methodology.

BACKGROUND: The incidence of lymph node metastasis is low in early gastric cancer. Early cancer without nodal metastasis may be theoretically eradicated by local treatment alone. However, except for gastrectomy, there exists no reliable treatment for submucosal cancer. We present a new laser system which irradiates the gastric wall from the serosal side combined with synchronous serosal cooling that can produce deep thermal coagulation without transmural damage. METHODS: A laser handpiece with a built-in surface cooling system has been devised, employing a diode laser of 980 nm wavelength. Animal experiments were conducted to ascertain whether this laser system could coagulate the mucosa and submucosa with acceptable injury to the muscular layer and serosa. The gastric wall was irradiated from the serosal side with synchronous serosal cooling at 6.0-10.0 W power output and 50-400 s exposure time. The degree and depth of damage were histologically assessed after 7 days. RESULTS: Ninety-one points were irradiated in 8 dogs. Mucosal coagulation was observed during the treatment while the serosa showed no serious injury. No gastric perforation occurred up to 7 days later. Histologically, the damage did not penetrate the gastric wall except for cases where 10.0 W and 400 s were used. Cauterization of the mucosa and submucosa with acceptable muscle layer damage was achieved in selected settings. CONCLUSIONS: This laser system enables thermal coagulation of the gastric mucosa and submucosa with acceptable muscle layer damage. This makes it a promising, novel, minimally invasive treatment for submucosal cancer.

Animals↗
PubMed1979

Effect of kallikrein-kinin system on ion transport across rat small intestine.

The flow of glucose and valine from the mucosal to the serosal side of rat intestine was stimulated significantly by kallikrein-kinin system. To elucidate the mechanism of this function the effects of kallidin on the transmural potential difference (PDt) between the serodal and the mucosal sides and the transepithelial resistance (Rt) of rat jejunum segment were investigated by electrophysiological procedures. The glucose-evoked PDt was not significantly altered by addition of kallidin to the mucosal fluid. Meanwhile, PDt which evoked by glucose was significantly shifted under the presence of 10-1-10(2) ng kallidin in the serosal fluid. The valine-evoked PDt was also changed by addition of 10 ng kallidin into the serosal fluid. However, the addition of glucose to the mucosal and kallidin to the serosal fluid did not affect on the Rt of the jejunum segment. These results suggest that kinin stimulated Na+ transport at the baso-lateral membrane of the intestinal absorptive cells, and the transports of other substances were enhanced consequently.

Animals↗
PubMedDate not supplied

Intraoperative radiation therapy for gastric cancer.

From among 115 patients who were treated by intraoperative radiation therapy (IORT), 94 patients were classified according to the histologic findings. The control 127 patients who were treated by operation alone during the same period were also histologically classified, and survivals were compared between the two groups. No difference in the survivals of patients with stage I was observed for the two groups. On the other hand, the survivals of patients with stages II through IV who were treated by IORT increased by nearly 10% to 20% at 5 years. A comparative study was also performed on the survivals between patients treated by IORT and those treated by operation alone according to the presence or absence of the serosal invasion and the grade of the lymph node metastasis. The number of patients treated by IORT in whom the serosal invasion and the lymph node metastasis were histologically examined was 57. The 171 control patients who underwent operation alone were examined histologically in the same fashion, and survivals for the two groups were compared. IORT did not afford any benefit if the lymph node metastasis was limited within n1 group or serosal invasion was not found. On the other hand, the 5-year survival rates for patients who were treated by IORT increased by nearly 10% when the serosal invasion was observed and by nearly 18% when n2 and n3 lymph node metastases were found.

Combined Modality Therapy↗
PubMed1993

Proposal of a risk score for recurrence in patients with curatively resected gastric cancer.

In this study, 168 patients who underwent curative resection for gastric cancer with prognostic serosal invasion [ps(+)] and 150 without prognostic serosal invasion [ps(-)] were analyzed separately to determine the prognostic importance of clinicopathological factors, and identify which patients were at high risk of recurrence. A multivariate analysis of survival time using Cox's proportional hazard model revealed the important prognostic factors to be: Lymph node involvement, the classification of gross appearance, macroscopic serosal invasion, and interstitial connective tissue in the ps(+) group; and lymph node involvement, macroscopic serosal invasion, and venous invasion in the ps(-) group. We proposed a risk score of recurrence based on the results of a further multivariate analysis called Hayashi's Quantification Analysis II, in which recurrence was chosen as an objective variable and the above prognostic factors were chosen as explanatory variables. Eighty-four percent of the patients with a score of 0 or higher in the ps(+) group and 83% of those with a score of +6 or higher in the ps(-) group showed recurrence. Thus, we believe that this score is useful for identifying those patients at high risk of recurrence, who should receive intensive chemotherapy even after curative resection.

Female↗
PubMed1981

Movement of thallium (I) ions in vitro.

1. Movement of thallium (I) ions across the mucosal epithelium of descending rat colon, stripped of the muscularis externa in vitro was determined under voltage clamp conditions. 2. The unidirectional fluxes of thallium (I) ions from mucosal to serosal side (M leads to S) and in the opposite direction (S leads to M) were measured at 0 mV. A net flux of thallium (I) ions was observed from serosal to mucosal side (0.216 nmoles . cm-2 . h-1) in other words, the mucosal epithelium is apparently asymmetric for the movement of thallium (I) ions. 3. The investigations were carried out on the range of -30mV up to +30 mV from mucosal to serosal side. Thallium (I) ions move from mucosal to serosal side (M leads to S) by diffusion only (Pi = 0.119 cm.h-1), whereas into the opposite direction (S leads to M) thallium (I) ions in addition to a voltage-dependent component are transported against an electrical gradient.

Animals↗
PubMed1995

Regulation of murine acid secretion by CO2.

To determine whether endogenous metabolic sources alone provide sufficient CO2 for acid secretion in mammals, basal and stimulated acid secretion and metabolic CO2 production were measured concurrently in mouse stomachs, in vitro, without exogenous CO2, and after addition of 5% CO2 serosally. Basal acid secretion was varied by changing luminal pH from 3.2 to 4.0. In the absence of an exogenous supply of CO2 acid secretion was stable under basal conditions and increased during cholinergic stimulation with carbachol. Serosal CO2 supply increased basal and stimulated acid secretion. The increase in basal acid secretion depended on the initial level of acid secretion. At pH 4.0, exogenous CO2 increased acid output (mean +/- SD) by 13% from 112 +/- 11 nmol/min to 126 +/- 8 nmol/min (P < 0.03), whereas at pH 3.6 the increase was 40% (63 +/- 14 to 88 +/- 20 nmol/min, P < 0.04) and 157% at pH 3.2 (21 +/- 13 to 54 +/- 14 nmol/min, P < 0.002). Following cholinergic stimulation a maximal acid output of 321 +/- 38 nmol/min was attained without serosal CO2, whilst addition of 5% CO2 to the serosal solution increased maximal acid secretion by 49% to 479 +/- 96 nmol/min (P < 0.005). Metabolic activity, measured as total gastric CO2 production, was greater as acid secretion rates increased [239 +/- 20 nmol/min at 21 +/- 13 nmol/min (luminal pH 3.2) versus 406 +/- 28 nmol/min at 321 +/- 17 nmol/min (after cholinergic stimulation)]. The data support the concept that basal and sub-maximal acid secretion can be maintained by CO2 available from metabolic sources, but full expression of the acid secretory apparatus requires exogenous CO2.

Animals↗
PubMed1984

Prognostic significance of intraperitoneal free cancer cells in gastric cancer patients.

We performed intraoperative peritoneal cytology in 171 gastric cancer patients undergoing curative surgery. Intraperitoneal free cancer cells were demonstrated in almost all patients in whom the area of serosal cancer invasion exceeded 15-20 cm2. In patients with both serosal cancer invasion and free cancer cells the 5-year survival rat was 13% as compared with 85% for patients who had neither, and 40% for patients who had serosal invasion but no free peritoneal cancer cells. Peritoneal metastasis was the most frequently observed recurrence pattern. Therefore, in gastric cancer patients with marked serosal invasion, intraoperative IP administration of cytocidal anticancer drugs should be considered.

Humans↗
PubMed1982

Asymmetric release of cyclic AMP from guinea-pig and rabbit gallbladder.

The release of cyclic adenosine 3':5'-monophosphate (cAMP) from guinea-pig and rabbit gallbladder was investigated in vitro. Serosal addition of prostaglandin E1 (PGE1) to luminally perfused guinea-pig gallbladders caused a concentration-dependent efflux of cAMP to the mucosal side, the threshold concentration of PGE1 being 10(-7) M. The efflux of cAMP to the serosal side was 7-fold lower. A mucosal sidedness of cAMP release was also observed in stripped preparations of rabbit gallbladder mucosa mounted between two half chambers. No cAMP was found in the solutions bathing the serosal layers isolated from rabbit gallbladders. Fluid secretion was observed at 10(-7) M PGE1, an effect mimicked by serosal, but not mucosal application of cAMP (3.3 x 10(-3) M). This is taken to indicate that the basolateral membrane is more easily permeated by cAMP than the apical membrane, since cAMP is believed to exerts its physiological effects from inside the cell. It is concluded that preferential release of cAMP to the mucosal side is not due to a higher permeability of the brush border membrane but rather represents an as yet undefined transport process which may be of importance for the regulation of excessive intracellular cAMP levels.

Alprostadil↗
PubMed1986

Absorption and secretion of potassium by rabbit descending colon.

Measurements of K fluxes under a variety of conditions have provided an internally consistent set of data that demonstrate active absorption and active secretion of K by rabbit descending colon in vitro. The properties of K diffusion across the paracellular pathway are those of a free solution shunt. With Na and Cl present on both sides of short-circuited tissues the two opposing active K transport systems balance each other, so that there is no net K transport. Net K absorption results when the transcellular secretory K flux is inhibited by 1. serosal addition of ouabain, 2. serosal addition of furosemide, or 3. omission of either Na or Cl from the serosal solution. Hence basolateral K uptake appears to be mediated by a furosemide-sensitive Na-Cl-K cotransport system in addition to the Na-K exchange pump. Luminal addition of mersalyl or orthovanadate inhibits active K absorption. The adenosine analogue 5'-N-ethylcarboxamide adenosine and the beta-adrenergic agent isoproterenol, added to the serosal solution, cause net K secretion which is inhibitable by furosemide. The secretory K fluxes, both under stimulated and nonstimulated conditions, are abolished by an opposing electrical gradient, suggesting conductive K exit across the apical cell membrane, whereas K absorption appears to be an electroneutral process.

Absorption↗
PubMed1987

Evidence for rheogenic sodium bicarbonate cotransport in the basolateral membrane of oxyntic cells of frog gastric fundus.

Ionic conductance properties of the basolateral cell membrane of oxyntic cells were studied in frog gastric fundus in vitro. After mounting the fundus in a modified Ussing chamber the serosal connective tissue was dissected off and individual oxyntic cells were punctured from the serosal surface with microelectrodes. Under resting conditions the membrane potential averaged -56.9, SD +/- 9.5 mV (n = 63), cytoplasm negative. Lowering or raising serosal HCO-3 concentration from 17.8 to 6 or 36 mmol/l respectively at constant PCO2 depolarized or hyperpolarized the cell membrane by +16.7 or -18.2 mV respectively. Sudden removal of serosal Na+ also depolarized the cell membrane (anomalous Nernst response). Since both the HCO-3 dependent and the Na+ dependent potential changes were strongly depressed by the disulfonic stilbene SITS and since the potential response to HCO-3 was virtually abolished in Na+-free solution we conclude that a rheogenic Na+ (HCO-3)n-cotransport system (n greater than 1) is present in the basolateral cell membrane of oxyntic cells. Its possible role in base transfer during HCl-secretion or HCO-3 secretion remains to be elucidated.

Animals↗
PubMed1988

Activation of chloride conductance induced by potassium in tracheal epithelium.

The effects of partial replacement of bathing solution sodium by potassium on potential difference, conductance and ion transport of canine tracheal epithelium were studied in Ussing chambers. Whereas up to 100 mmol/l mucosal K+ had no effect, raised serosal [K+] induced a concentration dependent decrease in transepithelial potential difference and increase in conductance. When serosal K+ was 100 mmol/l, the potential difference fell 30 mV to 1.1 +/- 1.0 mV and conductance rose 4.5 mS/cm2 to 6.6 +/- 0.9 mS/cm2. Seventy-five percent of the K+ induced conductance required Cl- (120 mmol/l) in the luminal bathing solution. Unidirectional Cl- fluxes were increased by raised serosal K+ but 14C-mannitol permeability was unchanged. The increased unidirectional Cl- flux induced by high K+ exposure was inhibited by luminal exposure to diphenylamine-2-carboxylic acid (DPC) or other chloride channel blockers, but was not inhibited by loop diuretics. These results suggest that in the presence of 100 mmol/l serosal K+ the transcellular chloride conductance of tracheal epithelium was increased. Increased chloride conductance of the apical cell membrane contributed to the raised transcellular permeability, but the route across the basolateral cell membrane was not identified.

Action Potentials↗
PubMed1979

A mechanistic explanation of the effect of potassium on goldfish intestinal transport.

Partial replacement of sodium by potassium or rubidium in the solution used to perfuse isolated intestinal segments of goldfish causes an increase in transmural electrical resistance. Serosal replacements have a stronger effect than mucosal replacements. A 70% inhibition of the glucose-evoked transmural electrical current is brought about by serosal replacement of 40 mM sodium by potassium. Transmural mucosal to serosal flux of 3-O-methyl-D-glucose is also strongly inhibited by serosal potassium. These inhibitory effects of potassium do not occur when the intestinal mucosa is stripped free from the intestinal muscular layers. It is concluded that potassium-induced muscular contractures cause a decrease in transport area by pressing the mucosal folds closer against each other. Certain effects of high potassium concentrations that have been reported in mammalian intestinal preparations may involve a similar mechanism.

Animals↗
PubMed1977

On the mechanism of the inhibitory effect of ethanol on intestinal glucose and water absorption.

Experiments were performed to investigate whether the inhibitory effect of ethanol on intestinal glucose transport is related to its action on the brush border or on the ATPase-dependent sodium pump of the basolateral membrane of the enterocyte. We compared the effect of ethanol on glucose and water transport when it was added either to the mucosal or to the serosal solution of an in vitro preparation of hamster jejunum. The purpose of the addition of ethanol to the serosal solution was to mimic a situation similar to that produced when ouabain is placed on the serosal side to inhibit the ATPase-dependent sodium pump at the basolateral membrane. The presence of 450 mM ethanol (2.07%) in the mucosal solution depressed glucose and water transport by 40 and 63%, respectively, but the presence of the same concentration of ethanol on the serosal side had no effect on glucose and water absorption. These findings seem to indicate that the depressing effect of ethanol on intestinal glucose and water transport cannot be ascribed to the inhibition of the Na+, K+-sensitive ATPase-dependent sodium pump located at the basolateral membrane.

Adenosine Triphosphatases↗
PubMed1981

Uptake of free and liposome-entrapped insulin by rat intestinal sacs in vitro.

Entrapment of insulin within distearoylphosphatidylcholine and cholesterol liposomes can protect the hormone from degradation in an in vitro gut-sac system and can increase the quantity of immunoreactive insulin that reaches the serosal fluid 26-fold. 95% of the insulin that reaches the serosal fluid is present within relatively intact liposomes. However, the presence of hydrolytic agents, probably enzymes, within the serosal fluid of everted gut sacs makes it necessary to separate the liposomes from the serosal fluid, by centrifugation, prior to release of the entrapped insulin with Triton X-100. This procedure prevents hydrolysis of the released insulin which can then be measured by radioimmune assay.

Animals↗
PubMed1977

Electrical properties and active solute transport in rat small intestine. I. Potential profile changes associated with sugar and amino acid transports.

Addition of D-glucose to the mucosal fluid resulted in a significant depolarization of the mucosal membrane potential (V-m) in rat duodenum, jejunum, and ileum accompanied by an increase in the transepithelial potential difference (PDt). On the other hand, L-glucose did not induce PDt and Vm changes. Glycine applied from the mucosal side also induced Vm-depolarization and PDt-increment in the ileum. Phlorizin added to the mucosal fluid or ouabain added to the serosal fluid inhibited the sugar-dependent changes in PDt and Vm. According to the analysis with an equivalent circuit model for the epithelium, it was concluded that an actively transported solute induced not only a depolarization of the mucosal (brush border) membrane but also a hyperpolarization of the serosal (baso-lateral) membrane of an epithelial cell, so that the origin of solute-induced PDt changes should be attributed to changes in emf's at both membranes. The hyperpolarization of the serosal membrane in the presence of an actively transported solute was attributed to a mechanism of serosal electrogenic sodium pump stimulated by the increase in the extrusion rate of Na+ co-transported into the cell with sugar or amino acid.

Animals↗
PubMed1978

Intracellular gradients of ion activities in the epithelial cells of the Necturus gallbladder recorded with ion-selective microelectrodes.

In Necturus gallbladder epithelial cells the intracellular electrical potential, as recorded with microelectrodes, varied from -28 mV in the mucosal end to about -50 mV in the serosal end of the transporting cell. The Na+ activity varied concurrently from about 39 mM to between 8 and 19 mM. Thus, within the cell both the recorded electrical and chemical gradients caused Na+ to move towards the serosal end. Serosal addition of ouabain (5 X 10(-4) M) caused the intracellular Na+ activity to attain electrochemical equilibrium within 30 min. However, the intracellular electrical potential gradient was only slowly affected. In cells from animals stored at 5 degrees C, the Cl- activity varied from about 55 mM in the mucosal end to 28 mM in the serosal end, and the K+ activity from 50 mM to between 95 and 131 mM. Both ions were close to electrochemical equilibrium within the cytoplasm but were too concentrated to be in equilibrium with the mucosal solution. Bubbling CO2 through the mucosal solution caused the intracellular gradients to vanish. When Na+ in the bathing solutions was exchanged for K+, the intracellular electrical potential became roughly constant at about -5 mV. The Cl- activity became constant in 65 mM, and the K+ activity became constant at 109 mM, both close to equilibrium with the mucosal solution. The Na+ activity was reduced to about 1 mM. The ratio of cytoplasmic resistivities between cells bathed in K+-rich saline to cells bathed in Na+-rich saline was measured by means of triple-barreled electrodes and compared to the same ratio as assessed from the activity measurements. The two values were equal only if one assumes the mobility of Na+ inside the cell to be less than 1/10 of the mobility of K+ or Cl-. The same conclusion was reached by comparing the intracellular Na+ flux calculated from the gradient of electrochemical potential to that flux assess from the net solute absorption. Animals kept at 15 degrees C had lower intracellular Na+ activities, higher Cl- and K+ activities, and higher rates of absorption than animals stored at 5 degrees C. Finally, the degree to which the intracellularly recorded electrical and chemical potentials could reflect an electrode artefact is discussed.

Animals↗
PubMed1984

Chloride secretion by canine tracheal epithelium: IV. Basolateral membrane K permeability parallels secretion rate.

We evaluated the K conductance properties of the basolateral membranes of the surface cells of canine tracheal epithelium using microelectrode techniques. Studies were conducted under basal conditions (indomethacin, 10(-6) M, mucosal solution) and after stimulation of electrogenic Cl secretion with epinephrine (10(-6) M, serosal solution). Elevated serosal solution [K] depolarized the electrical potential differences across the apical (psi a) and basolateral (psi b) membranes in both the presence and absence of epinephrine. Serosal barium (0.5 mM) also depolarized psi a and psi b and selectively increased basolateral membrane resistance threefold. We also used K-selective microelectrodes to determine cell K activity (acK) and the driving force for K transport across the limiting membranes under basal and stimulated conditions. Stimulation of Cl secretion was not associated with significant changes in psi b or acK so that the driving force for K exit from cell to serosal solution (ca. 20 mV) was not altered. There was close agreement between the basolateral membrane electromotive force (Eb) determined from prior studies (M.J. Welsh, P.L. Smith and R.A. Frizzell, J. Membrane Biol. 71:209-218, 1983) and the chemical potential difference for K across this barrier (EbK) in the presence and absence of epinephrine. These findings support the notion that the basolateral membrane is characterized by a high conductance to K under both secreting and nonsecreting conditions and indicate that the decrease in basolateral membrane resistance that accompanies stimulation of Cl secretion results from an increase in its K conductance. This obviates changes in acK that would otherwise accompany increased Na/K pump activity and, by hyperpolarizing psi a, establishes the electrical driving force for Cl secretion across the apical membrane.

Animals↗