Search PubMed⌕ Search

Biomedical subjects

D Pansu

Publications and source records attributed to D Pansu.

At least 37 records · Page 2Linked to original sources

[Effect of C-terminal derivatives of sorbin on ileal ion transport stimulated by VIP in rats].

OBJECTIVES AND METHODS: Sorbin, a peptide isolated from porcine intestine and composed of 153 aminoacids, has been purified because its specific action is to increase water and ion absorption in the intestine and the gall bladder. We showed that synthetic peptides containing the amidated C-terminal part of sorbin had the same activity as the natural molecule in increasing duodenal absorption. In order to characterize the site of action of sorbin, the effect of two C-terminal derivatives were determined in ileal ligated loops in situ in anaesthetised rats, following VIP-induced water and electrolyte secretions. Their effect was compared to those of metenkephalinamide, NPY and somatostatin. Unidirectional fluxes were studied to analyze the mechanism of action of sorbin, by means of 22Na, administered into the intestinal loop, and 36Cl, injected into blood. RESULTS: Results show that C20-sorbin and C7-sorbin decreased the VIP-stimulated net flux of water (inhibition of 40 and 37%, respectively), Na (inhibition of 31 and 30%), C1 (inhibition of 80 and 63%) and HCO3 (inhibition of 15 and 25%). These effects are evidently greater than those produced by equimolar doses of NPY, somatostatin, and 32 times higher dose of metenkephalinamide. Sorbin acts as a potent anti-secretor, anti-VIP, in rat ileum.

Animals↗

Comparative duodenal, jejunal and ileal responses to luminal saline load.

Intestinal ionic exchanges were studied in rat duodenal, jejunal and ileal ligated loops in response to different luminal saline loads: NaCl concentration varied from 150-0 mM, solutions being made isoosmotic with mannitol. The contact delay was 60 min. An exponential relationship was found between water, Na and Cl movements and the initial saline concentration. Maximal absorption was obtained with 150 mM NaCl, and was significantly higher in the duodenum than in the jejunum and ileum. The NaCl concentration for which water, Na, and Cl movements were null was approximately 70 mM NaCl in the duodenum and jejunum, 41 mM for Na and 18 mM for Cl in the ileum. The water efflux induced by the 0-mM NaCl test solution was maximal in the duodenum (1.5 +/- 0.2 ml/h) and decreased in the jejunum (0.8 +/- 0.1 ml/h) and ileum (0.3 +/- 0.1 ml/h) as did sodium, chloride and non-chloride anion efflux. These data support the functional heterogeneity of the small intestine regulating the water and ion exchange in response to luminal saline load, the main difference being connected with the efflux capacity of the mucosa, decreasing from the duodenum to the jejunum and ileum.

Animals↗

Solubility and intestinal transit time limit calcium absorption in rats.

Six-week-old male rats were placed on two high calcium regimens: one with calcium carbonate and monobasic calcium phosphate, with calcium content increased via calcium carbonate; and another with calcium phosphate and calcium gluconate, with calcium gluconate the source of increased calcium. Animals fed the gluconate-containing diets absorbed 29% of the ingested calcium over the entire calcium intake range, whereas those fed the calcium carbonate diets absorbed 25% over an intake range of 225 to 450 mg Ca/d, but at calcium intakes above 450 mg Ca/d their absorption reached a plateau at approximately 109 mg/d. Active calcium transport decreased with increased calcium intake in both the calcium carbonate- and calcium gluconate-fed groups. Nonsaturable transport was unchanged as a result of increasing calcium intake and did not differ among the diet groups. Because the absorptive processes were unaffected by the calcium source, events in the lumen must have been responsible for the observed differences. Because phosphate is nearly 18 times more soluble than carbonate, very little calcium of calcium carbonate origin can have been solubilized in the presence of phosphate and this, we conclude, accounts for the limit on calcium absorption observed in diets high in calcium carbonate. Moreover, when intake is expressed as soluble calcium, absorption approaches 50%, the value expected when intestinal transit time (approximately 3 h) is multiplied by 16%/h, the experimental value of nonsaturable absorption.

Animals↗

Effect of sorbin on duodenal absorption of water and electrolytes in the rat.

Sorbin is a newly isolated intestinal peptide that has been purified because of its ability to induce water absorption. The effects that sorbin and some synthetic peptides corresponding to its C-terminal sequence have on duodenal absorption of water, chloride, and sodium were studied in comparison with the effects of vasoactive intestinal peptide (VIP), [D-Ala,Met]-enkephalinamide (DAMA), and angiotensin II. The technique of an in situ ligated duodenal loop in the rat was used for all peptides. Under the experimental conditions used, a low basal secretion of water, chloride, and sodium was obtained; VIP induced an increase of the secretion, whereas DAMA induced an absorption, both in the nanomolar dose range. Angiotensin II and sorbin induced an absorption in the picomolar dose range. The most effective doses of sorbin peptides but not of angiotensin induced the lowest final concentrations of Na+ and Cl- obtainable without inducing secondary water secretion. All synthetic peptides containing the C-terminal heptapeptide of sorbin were active in the picomolar dose range. Contrary to angiotensin, they had no effect on blood pressure.

Amino Acid Sequence↗

Comparison of VIP-induced electrolyte secretion at three levels in rat small intestine.

Duodenal, jejunal and ileal loops were prepared and an iso-osmotic test solution injected, containing 80 mM Na+, 5-mM K+, 1.2 mM Ca2+, 77 mM Cl-, 10 mM HCO3- and 136 mM mannitol. 14CPEG 4000 was used as a non-absorbable marker and 36Cl was added to measure the bidirectional fluxes. During the 60-min in vivo incubation time, the duodenum actively secreted bicarbonate, a virtually zero flux in the jejunum was observed, whereas the ileum absorbed water and chloride and secreted bicarbonate. The response to the perfused doses of 0.15 to 2.4 nmol.100 g-1.h-1 of VIP (vasoactive intestinal peptide) differed qualitatively and quantitatively in the 3 segments: VIP increased bicarbonate secretion and induced chloride secretion in the duodenum, induced chloride secretion in the jejunum without changing bicarbonate minimal influx, induced bicarbonate secretion and suppressed chloride absorption in the ileum. The minimal dose required was lower in the duodenum (0.3 nmol.100 g-1.h-1) than in the jejunum and ileum (1.2 nmol.100 g-1.h-1). The functional heterogeneity of the small intestine was clearly demonstrated after VIP stimulation.

Animals↗

Isolation and characterisation of porcine sorbin.

Sorbin has been isolated from extracts of porcine upper intestine, and the biological activity in absorbing water and electrolytes utilized to monitor the purification procedure. Pure sorbin was obtained in a yield of about 1 mg/Mg boiled intestine. The protein chain has 153 amino acid residues and the primary structure was determined by analyses of CNBr-cleaved fragments and four enzymatic digests. The protein has a free N-terminal Met and an amidated C-terminal Ala. No structural similarity was observed with other known proteins in data bases, but several segments have special properties and the C-terminal half is rich in Pro and Arg.

Absorption↗

Effect of D ala2 metenkephalinamide on feline jejunal and ileal water and electrolyte transport.

The aim of this investigation was to compare the effect of an opioid, D ala2 metenkephalinamide (DAMA), on net jejunal and ileal water and electrolyte fluxes using the gut perfusion technique in the anesthetized cat. Intestinal transport was measured during intravenous infusion of serial doses of 2, 6, and 18 micrograms.kg-1.h-1 of DAMA in 6 cats. Each cat was its own control during an intravenous infusion of 150 mmol/l NaCl preceding the first dose of peptide and following the last dose of DAMA. Both jejunal and ileal segments were isolated by inflated balloons and were studied at the same time. Fifteen ml of an iso-osmolar test solution with hypo-osmolar ion contents and complementary mannitol were administered in the upstream tube and collected 1 h later in the downstream tube. In the jejunum, water secretion was dose-dependently reversed to an absorption from a control value of +0.5 +/- 0.4 to -0.83 +/- 0.5 ml.h-1.10 cm-1; in the ileum, water absorption was increased from -0.5 +/- 0.3 to -1.5 +/- 0.2 ml.h-1.10 cm-1. The net absorption of all electrolytes, ie sodium, chloride, bicarbonate, potassium and calcium also increased during peptide administration. However, a qualitative difference in the ion transport was observed between the jejunum and the ileum.

Animals↗

Theophylline inhibits transcellular Ca transport in intestine and Ca binding by CaBP.

Theophylline, when added to the incubation medium of everted duodenal sacs prepared from rats on a low-calcium diet, was found to inhibit transcellular Ca transport in a concentration-dependent manner, with an inhibitor constant (Ki) of 10.8 mM theophylline. Neither the rate of cellular Ca entry, as evaluated with the aid of brush-border membrane vesicles, nor the rate of cellular Ca extrusion, assessed by measuring ATP-dependent Ca uptake of basolateral membrane vesicles, was significantly altered by the addition of theophylline to the uptake media. However, Ca-binding by calcium-binding protein (CaBP; calbindin D9k), Mr approximately 8,800) was depressed by theophylline in a concentration-dependent manner, with Ki = 3.2 mM theophylline. Theophylline had no effect on Ca binding by calmodulin and the theophylline-induced inhibition of transcellular calcium transport was independent of adenosine 3',5'-cyclic monophosphate levels. Theophylline also had no effect on paracellular Ca movement. Since the theophylline-induced inhibition of Ca-binding by CaBP paralleled the inhibition of transcellular Ca transport, it is concluded that CaBP functions in transcellular Ca transport via its ability to bind Ca.

Animals↗

Circannual and circa-hemiannual rhythms of basal and stimulated gastric secretion in conscious cats.

Conscious cats equipped with a gastric fistula and a denervated Heidenhain pouch were submitted to weekly measurements of the basal and pentagastrin-stimulated gastric secretion for 1 to 14 years. Rhythms of basal secretion were documented in 37 cats for the group studies, in 25 cats only for the individual studies which required at least whole year data. Twelve-month or 6-month rhythms were detected for each variable studied, i.e. volume, acid, pepsin, fucose and uronic acid outputs in the group studies, with peaks for volume, acid and pepsin in Winter, peaks for uronic acid in Spring and Fall indicating different rhythms for oxyntic, chief and mucous cells. Individual studies detected rhythms in 25% of the analyses, and demonstrated male and female and cat to cat differences. Spectral analysis in 3 cats confirmed the differences in the individual rhythms with prominent peaks differing from 365 days in 50% of the cases. Chronopharmacological responses to pentagastrin were documented for volume, acid and pepsin outputs in 5 male and 6 female cats. Group analysis detected a Winter acrophase for volume and acid secretion and a Summer acrophase for pepsin secretion. Analysis of the stimulated response data showed interindividual variation but a higher percentage of detection for rhythms, i.e. 38% for all variables and 50% for pepsin secretion. Different rhythms in acid and pepsin secretion documented in individual studies could provide the basis of a better understanding of the discrepancies reported in the literature concerning the seasonal incidence of peptic ulcer disease.

Animals↗

An analysis of intestinal calcium transport across the rat intestine.

Kinetic analysis of transmural calcium transport, as evaluated by in situ intestinal loops, has confirmed the existence of two transport processes, a saturable, transcellular one that is regulated by vitamin D and predominates in the proximal intestine and a nonsaturable process similar in intensity throughout the intestine. Transport data obtained from everted sac experiments are kinetically consistent with events in the in situ loop. Analysis of the three component steps making up the saturable process, i.e., entry across the brush-border membrane, intracellular diffusion, and extrusion across the basolateral membrane, indicates that intracellular diffusion is likely to be the limiting step. Active calcium transport varies directly and proportionately with the content of calcium-binding protein (CaBP), a specific molecular expression of the action of vitamin D. Since CaBP is a cytosolic protein, it may act to facilitate calcium diffusion, a proposition advanced by Kretsinger, Mann, and Simmons and supported here quantitatively. We calculate that the rate of intracellular calcium diffusion in the absence of CaBP is only approximately 1/70 of what is found in the vitamin D-replete cell. Similar considerations have led to the proposal that calcium moved by the nonsaturable process travels largely via the paracellular route. The kinetic parameters derived here, i.e., Vm = 22 mumol X h-1 X g (wt wt-1, Km = 3.9 mM, and a nonsaturable rate of 0.16/h, can be used to predict calcium absorption data as determined in previously published balance experiments.

Aging↗

Localization of vitamin D-dependent active Ca2+ transport in rat duodenum and relation to CaBP.

Vitamin D-replete (+D) and vitamin D-deficient (-D) rats received by intraperitoneal injection varying amounts of 1,25-dihydroxyvitamin D3, and 4 h (+D) or 9 h (-D) later everted duodenal sacs were prepared to evaluate active calcium transport, i.e., the amount of calcium found in the serosal fluid. At the same time, duodenal calcium-binding protein (CaBP) content was measured. Calcium transport was a close positive function of CaBP content. It was not detectable when CaBP content was zero and increased linearly without plateauing as CaBP content increased to 100 nmol calcium bound/g mucosa. Trifluoperazine (TFP) inhibited active calcium transport in a concentration-dependent manner. Experiments using vesicles prepared from brush-border or basolateral membranes indicated that TFP inhibited the calcium-extrusion process, with virtually no effect on calcium entry. It is concluded that vitamin D exerts its major regulation of active calcium transport in the rat duodenum via CaBP on transport steps beyond brush-border entry.

Animals↗

[Intestinal absorption of calcium and its regulation. Tissue, membrane and molecular events].

The intestinal absorption of calcium involves an active transport against an electrochemical gradient, a saturable and a nonsaturable transfer following the gradient. The active and the saturable components are transcellular, the nonsaturable component is partly paracellular. Calcium transfer through the intestinal cell includes three steps: 1) the "down-hill" crossing of the brush-border implies binding to specific sites, carrier-mediated transport using channels or carrier proteins specific to Ca and dependent upon composition phosphorylations alkaline phosphatases; the phospholipid composition of the brush-border also plays a role; 2) the intracellular transfer is characterized by an uptake by such organelles as mitochondria, lysosomes and Golgi vesicles and by a transfer on a specific calcium-binding protein; 3) the "up-hill" transfer across the baso-lateral membrane requires energy and energy is mediated by an ATP-activated Ca2+ pump and a Na+/Ca2+ antiport. 1.25 dihydroxycholecalciferol, steroid hormone synthetized from vitamin D3 is the major direct regulator of Ca absorption: in the vitamin D-deprived animal, it increases the selective permeability for Ca of the brush-border, induces the synthesis of proteins after genomic transcription, activates the Ca-ATPases, and acts as a trophic hormone. The other hormones principally act by modulation of the renal biosynthesis of 1.25 dihydroxycholecalciferol. The efficiency of Ca absorption depends on site, with duodenum greater than jejunum greater than caecum greater than ileum. Dietary constituents such as carbohydrates and amino acids increase Ca absorption whereas phytic acid and excess of phosphorus decrease it. They act by modifying Ca bioavailability and perhaps brush-border permeability. Adaptation to increased demand occurs during growth, pregnancy and lactation in normal states but disappears during vitamin D deficiency. In man, lowered efficiency with increasing age is often aggravated by a low calcium diet.

Animals↗

Calcium uptake by isolated rat intestinal cells.

Intestinal cells were isolated by a combination of mechanical and enzymatic means, and their calcium uptake was assayed by a rapid filtration procedure. Calcium uptake was a time- and concentration-dependent process that was markedly elevated at 25 and 37 degrees C, as compared to 0 degree C. Cells isolated from rat duodenum exhibited higher uptakes than cells from jejunum, which in turn took up more calcium than cells from the ileum. Duodenal cells from vitamin D-deficient animals took up less calcium than cells from vitamin D-replete cells. In vivo vitamin D repletion with 1,25-dihydroxyvitamin D3 raised calcium uptake by duodenal cells from treated animals toward that of cells from replete rats. Furthermore, calcium uptake by duodenal cells from vitamin D-deficient animals approximated that of ileal cells from replete rats. These findings with isolated cells parallel prior findings of tissue calcium transport and suggest that cellular calcium uptake may be related to the saturable component of intestinal calcium absorption. Isolated intestinal cells may therefore constitute one experimental model for the study of transcellular calcium transport.

Animals↗