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Functional significance of histologic alterations induced by Escherichia coli pig-specific, mouse-negative, heat-stable enterotoxin (STb).

In contrast to cholera enterotoxin and other Escherichia coli enterotoxins, a pig-specific, heat-stable E. coli enterotoxin (STb) causes morphologic lesions (loss of villous epithelial cells and partial villous atrophy). These lesions reflect a loss of absorptive cells and thus suggest that STb causes impaired absorption as well as inducing net secretion. The present studies assess functional significance of morphologic changes induced by STb. Net fluid movement, mucosal surface area, sucrase activity and the electrical response induced by alanine were measured in swine jejunal loops exposed to E. coli culture filtrates with and without STb. Net fluid secretion (-11.1 +/- 1.1 ml) occurred in some STb loops (secretors) and net absorption (2.7 +/- 0.3 ml) in others (nonsecretors), but net absorption occurred in all control loops (4.9 +/- 0.2 ml). The mucosal surface area of STb loops was about 20% less than that of controls (P less than 0.01). Sucrase activity was also lower (about 15%) in STb loops than in control loops (P less than 0.01). The electrical response induced by alanine in mucosa from nonsecreting STb loops did not differ from that induced in mucosa from control loops. However, the response to alanine in mucosa from secreting STb loops was reduced about 70% from that in mucosa from nonsecreting STb loops or from control loops (P less than 0.05). It is concluded that reduced sucrase activity is a functional correlate to villous atrophy induced by STb, that STb impairs alanine absorption in some loops (secretors), and that the impaired alanine absorption is independent of the decreased surface area caused by STb. Because the impaired alanine absorption occurred independent of the decreases in surface area, it is suggested that the secretory response to STb is associated with an impairment of active absorption of alanine.

Animals↗

Serosal application of Ba(2+) induces oscillatory chloride secretion via activation of submucosal cholinergic neurones in guinea-pig distal colon.

The enteric nervous system regulates ion and fluid secretion in the mammalian intestine at both resting and stimulated conditions. To determine the type and activation mechanism of neurones involved, mucosa-submucosa sheets isolated from guinea-pig distal colon were studied in vitro in Ussing chambers. Serosal addition of 0.5-1 mM barium (Ba(2+)), a potassium (K(+)) channel inhibitor, caused oscillatory increases in short-circuit current (I(sc)). Mean values of the size and frequency of I(sc) were 369.1 microA cm(-2) and 2.3 min(-1). The oscillatory I(sc) induced by the low concentrations of Ba(2+) was blocked by either higher concentrations of Ba(2+) (2-5 mM) or other K(+) channel inhibitors, such as tetraethylammonium (TEA) (1 mM) and quinine (20 mM). The Ba(2+)-induced oscillatory I(sc) was also inhibited by tetrodotoxin (TTX) and atropine. In a nominally Ca(2+) free solution plus serosal addition of 0.1 mM ethylene glycol-bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), a Ca(2+) chelator, the oscillatory I(sc) slowed and diminished. Further, the Ba(2+)-induced oscillatory I(sc) was partially inhibited by apical addition of 100 microM 5'-nitro-2-(3-phenylpropylamino)benzoic-acid (NPPB), a Cl(-) channel inhibitor, and completely disappeared in a low Cl(-) solution (11 mM) on both sides. On the other hand, application of either cimetidine, a histamine H(2) receptor antagonist, or hexamethonium, a nicotinic antagonist, to the serosal side did not affect the Ba(2+)-induced oscillatory I(sc). In conclusion, the Ba(2+)-induced oscillatory I(sc) is the transepithelial Cl(-) current which is stimulated by activation of cholinergic neurones in submucosal plexus of guinea-pig distal colon.

Animals↗

Differential bile-pancreatic secretory effects of CCK-58 and CCK-8.

In this work, we 1) synthesized rat CCK-58, 2) determined the amounts and forms of rat CCK in whole blood after stimulation of its release by casein, 3) determined the potency of CCK-8 and CCK-58 peptides to displace labeled CCK-8 from CCK(A) and CCK(B) receptors transfected into Chinese hamster ovary (CHO) cells, and 4) examined the biological actions of CCK-8 and rat CCK-58 in an anesthetized rat model. CCK-58 was the only detected endocrine form of CCK in rat blood. Synthetic rat CCK-58 was less potent than CCK-8 for displacing the label from CCK(A) and CCK(B) receptors in transfected CHO cells. However, rat CCK-58 was more potent than CCK-8 for stimulation of pancreatic protein secretion in the anesthetized rat. In addition, CCK-58 but not CCK-8 stimulated fluid secretion in this anesthetized rat model. These data suggest that regions outside the COOH terminus of rat CCK-58 influence the expression of CCK biological activity. The presence of only CCK-58 in the circulation and the fact that its biological activity differs from CCK-8 suggests that CCK-58 deserves scrutiny in other physiological models of CCK activity.

Amino Acids↗

Expression of CFTR and a cAMP-stimulated chloride secretory current in cultured human fetal alveolar epithelial cells.

During development the fetal lung secretes fluid that is osmotically linked to chloride (Cl-) transport. One possible pathway for Cl- secretion across the fetal pulmonary epithelium is through the cystic fibrosis transmembrane conductance regulator (CFTR). CFTR is expressed in epithelia and functions as a Cl- channel regulated by cyclic adenosine monophosphate (cAMP)-dependent protein kinase and intracellular ATP. Previous studies have shown that CFTR mRNA is expressed throughout the human fetal pulmonary epithelium and CFTR protein can be immunoprecipitated from human fetal lung homogenates. In cultured fetal lung tissue explants, CFTR mRNA was localized to alveolar epithelial cells. To test the hypothesis that fetal alveolar epithelial cells express functional CFTR, we immunolocalized CFTR in human fetal lung and looked for evidence of Cl- secretion in cultured alveolar epithelial cell monolayers. Monoclonal anti-CFTR antibodies localized CFTR in cultured lung explants to the epithelial cells, predominantly at the apical surface. Bioelectric properties of cultured monolayers of midgestation fetal alveolar epithelial cells were measured in modified Ussing chambers. In unstimulated monolayers, transepithelial electrical potential difference (psi t) = -1.1 +/- 0.1 mV, transepithelial resistance (Rt) = 768 +/- 58 omega.cm2, and short-circuit current (Isc) = 1.9 +/- 0.2 microA/cm2 (mean +/- SE, n = 17). Addition of amiloride to the apical surface significantly decreased basal Isc. Apical diphenylamine-2-carboxylate (DPC), a Cl- channel inhibitor, caused no significant change in basal Isc. In the presence of apical amiloride, isoproterenol significantly increased Isc, a response that was inhibited by apical DPC and submucosal bumetanide. The cAMP agonists forskolin and IBMX also stimulated Isc.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗

Inhibition of gastric secretion in guinea pig by relatively low dose ionizing radiation.

We evaluated the effect of a single dose of ionizing radiation on gastric secretion in awake guinea pigs equipped with a permanent gastric cannula. Changes in gastric secretion were measured using a dye dilution technique. Infusion of histamine increased acid and fluid output and there was a positive correlation (r = 0.93) between the two. Total body irradiation with 400 cGy, like cimetidine, suppressed acid and fluid secretion under basal conditions and during histamine stimulation by 50-90%. Recovery from the radiation damage was only partial after one week. Irradiation inhibited the rise in gastric juice volume during histamine stimulation and also reduced the normal gain in body weight of the guinea pig. These results demonstrate that ionizing radiations have an immediate and long lasting effects on the gastric mucosal function of the guinea pig.

Animals↗

Endometrial secretion aspiration prior to embryo transfer does not reduce implantation rates.

Analysis of protein patterns in endometrial secretion fluid may offer a relatively non-invasive means of assessing endometrial receptivity during fertility treatment cycles. In order to study the impact of the removal of endometrial secretions on embryo implantation, a prospective matched controlled study was performed. In 66 women undergoing IVF, endometrial fluid was obtained transcervically by aspiration just prior to embryo transfer (study group). Biochemical and ongoing pregnancy rates were compared with 66 control patients matched for stimulation treatment protocol, age, number of collected oocytes and number of high quality embryos. The protein content and uterine fluid protein profile in each sample was determined. Respective biochemical and ongoing pregnancy rates per embryo transfer were 36 and 33% in patients who underwent aspiration of endometrial secretion, compared with 33 and 30% respectively in matched control patients (P = 0.84 and P = 0.85). The protein content in endometrial fluid was sufficient for protein pattern analysis. Uterine fluid aspiration prior to IVF embryo transfer is a safe method for obtaining sufficient material for uterine secretion electrophoresis, thus allowing analysis of protein patterns serving as receptivity markers during treatment cycles. This technique may offer a novel tool for assessing endometrial receptivity during treatment cycles without affecting implantation rates.

Adult↗

Electrophoretic characterization of proteins in oviduct fluid of cows during the estrous cycle.

This study was undertaken to examine the in vivo protein composition of the bovine oviduct during the estrous cycle. Oviduct fluid was collected daily from four oviduct-cannulated dairy cows for a total of four complete estrous cycles. Fluid secretion followed a definite cyclic pattern, with maximum secretion occurring at estrus in all cycles. Protein concentration fluctuated during the cycle and varied among animals. In general, protein concentration was lower at the time of estrus. Total protein in oviduct fluid, however, was higher around estrus, indicating increased transudation or secretion by the oviduct. One-dimensional SDS-PAGE separation revealed the protein pattern of oviduct fluid to be generally similar to that of blood serum and follicular fluid. Two proteins appeared to be oviduct-specific. The first, a protein of approximately 47 kDa, was evident in oviduct fluid throughout the estrous cycle. The second protein, evident as a broad diffuse staining band above albumin, appeared for only 3-4 days at or near ovulation. This protein had a molecular weight of 80-95 kDa and stained positive for carbohydrate with periodic acid-Schiff reagent. These studies indicate that the in vivo protein composition of oviduct fluid varies with the estrous cycle, and that around estrus, an oviduct-specific glycoprotein is present.

Animals↗

Sympathetic control of jejunal fluid and electrolyte transport. An experimental study in cats and rats.

The study was performed to test the hypothesis that the sympathetic nerves to the small intestine might be involved in the feed-back control of extra-cellular fluid volume. In a series of experiments in anaesthetized cats, we studied how efferent stimulation of the splanchnic nerves affected jejunal fluid transport rate. The mechanisms behind the observed responses were analysed and we also attempted to determine the physiological role of the sympathetic supply to the jejunal mucosa. Splanchnic nerve stimulation (SNS) led to an increase in net fluid absorption provided that glucose was present in the jejunal lumen. In segments exposed to cholera toxin, SNS had a very pronounced inhibitory effect on net fluid secretion. The absorptive response was significant at a stimulation rate of 2 Hz and the maximal response occurred at 4-8 Hz. The effect was blocked by phentolamine, an alpha-adrenergic antagonist, and it was mimicked by a close intraarterial infusion of noradrenaline, an alpha-adrenergic agonist. The absorptive response to SNS was still present after blockade of the intestinal vasoconstrictor nerves with hexamethonium, indicating that the sympathetic pathways to the epithelium and to vascular smooth muscle may employ different ganglionic transmitter mechanisms. The increase in net fluid absorption induced by SNS was due to the inhibition of a secretory process. The response was not secondary to changes in intestinal blood flow. The effect of noradrenaline infusion was abolished after blockade of the intestinal nerves with tetrodotoxin, a finding which indicates that the sympathetic nerves may act by modulation of the activity in intramural secretory nervous pathways. Unloading of carotid sinus baroreceptors led to an increase in jejunal fluid absorption, a response which was abolished by division of the sympathetic fibres to the segment. Unloading of cardiac mechanoreceptors enhanced jejunal fluid absorption or decreased cholera secretion, effects which were abolished by previous cervical vagotomy or local alpha-adrenoreceptor blockade of the jejunal segment. Afferent stimulation of the right cardiac nerve led to an inhibition of net fluid absorption if the nerve was stimulated with high-threshold parameters at frequencies below 1 Hz.

Adrenergic alpha-Agonists↗

Rat pancreas adenylate cyclase V. Its presence in isolated rat pancreatic acinar cells.

(1) In order to determine the cellular localization of the secretin- and pancreozymin-sensitive adenylate cyclase in rat pancreas, the occurence of this enzyme system has been investigated in isolated pancreatic cells. (2) Digestion of rat pancreatic lobules with collagenase yields a preparation of isolated cells which upon differential morphological analysis appears to consist for 97% of acinar cells and to contain for fewer centro-acinar and ductal cells than undissociated lobules. (3) Expressed per mg protein, the isolated cells contain the same amount of DNA, chymotrypsin and lactic dehydrogenase as the undissociated tissue. The stimulated adenylate cyclase activity is nearly entirely recovered in the isolated acinar cells, as is also the case for the low Km adenosine 3',5-cyclic monophosphate phosphodiesterase activity and the adenosine 3',5'-cyclic monophosphate (cyclic AMP) content. Marked losses are noted for the basal adenylate cyclase and the high Km cyclic AMP phosphodiesterase activities. (4) Washing the isolated acinar cells in Krebs-Ringer bicarbonate medium containing 10 mM 1-methyl-3-isobutylxanthine causes a cyclic AMP level 2.6 times that in cells washed in Krebs-Ringer bicarbonate alone. The cyclic AMP level is further increased by subsequently incubating the cells for 10 min in the presence of 3-10(-7) M pancreozymin-C-octapeptide or secretin to values 1.7 or 4.7 times the control level in cells incubated for 10 min with 1-methyl-3-isobutylxanthine alone. (5) It is suggested that the adenylate cyclase of the acinar cells may be involved, with another factor, in the stimulation of enzyme secretion, whereas a ductular cyclase would function in the regulation of the bicarbonate-dependent fluid secretion.

3',5'-Cyclic-AMP Phosphodiesterases↗

Immunoglobulin A secretion into pancreatic juice as a novel marker of local immune defense and exocrine pancreatic function.

The importance of secretory immunoglobulin A (IgA) of local immune defense in the gastrointestinal tract has gained increasing acceptance. Bacterial contamination is a major factor related to mortality in acute pancreatitis. However, very little is known about IgA in pancreatic juice. Pure pancreatic juice was collected from 40 patients undergoing pancreatoduodenectomy. The patients were divided into three groups according to the degree of preoperative pancreatic duct obstruction, as follows: normal, narrowed, and obstructed. IgA concentration, amylase activity, and daily volume of pancreatic juice were measured. Daily IgA secretion into pancreatic juice was constant during the early period after the operation. The concentration of IgA in the control group was 5 +/- 0.8 microg/ml, and IgA daily secretion was 1.2 +/- 0.2 mg/day. Pancreatic duct obstruction resulted in a marked decrease in both amylase and pancreatic juice secretion. The concentration of IgA, however, was markedly increased in the narrowed group (11.1 +/- 2.4 microg/ml) and the obstructed group (32.5 +/- 5.4 microg/ml). The concentration of amylase increased with the increase in pancreatic juice. Conversely, the concentration of IgA increased with the decrease in volume of pancreatic juice. Similarly, the increased in IgA concentrations positively correlated with the decrease in amylase activity. In conclusion, the mechanisms that modulate IgA secretion in the human pancreas are essentially different from those that modulate digestive enzyme and fluid secretion. IgA in pancreatic juice may play an important role in pathological conditions such as pancreatic duct obstruction. As such, the measurement of IgA in pancreatic juice may potentially be used as a new marker of local immune defense and exocrine pancreatic function.

Amylases↗

Calcium secretion from the feline pancreas. Influence of hormonal and cholinergic secretagogues and of serum calcium.

The influences of secretagogues and of elevated serum calcium concentrations on the calcium secretion from the cat pancreas have been studied in vivo. During a high and constant fluid secretion rate evoked by a background infusion of secretin, additional infusions of both cholecystokinin-pancreozymin and urecholine led to a dose-dependent increase in calcium secretion in pancreatic juice parallel to the rise of protein. The amount of calcium in pancreatic juice associated to 1 mg protein (18.3 nmol/mg protein) calculated from regression analysis was independent of dose or kind of stimulus used. The protein-independent pancreatic juice calcium fraction was 0.184 mM in normocalcemia. During an episode of hypercalcemia produced by an intravenous calcium infusion, the protein-independent calcium fraction was increased and correlated linearly to the serum calcium concentration. We conclude that pancreatic juice calcium consists of two major fractions, one being associated with the enzyme protein and stimulated by secretagogues, and the other being protein independent and directly dependent on the extracellular calcium concentration.

Animals↗

Ultrastructure of the secretion of prostasomes from benign and malignant epithelial cells in the prostate.

BACKGROUND: Prostate epithelial cells are producing, among other things, a fluid secretion containing small bodies, the prostasomes. The mechanism of synthesis of the prostasomes is not known in details, neither is it known whether the mode of prostasome production changes at a neoplastic transformation of the prostate cells. Due to the small size of the prostasomes, we have used electron microscopy for evaluating the production and distribution of prostasomes in benign and neoplastic cells of the prostate. METHODS: Benign and neoplastic areas in plastic embedded core biopsy specimens of prostate tissue were identified, and secreting cells were selected. The corresponding areas on the plastic blocks were further processed for examination in the electron microscope. RESULTS: The electron microscopical examination showed that the secretory machinery was similar in both types of tissue. Thus, in both benign and well-differentiated neoplastic cells studied, the formation of storage vesicles in the Golgi areas was similar, the content of the vesicles appeared similar, the structure and distribution of prostasomes were alike, and in both benign and malignant tissue, the secretion in the gland ducts showed the same appearance with many prostasomes. CONCLUSION: We conclude that cells in benign prostate tissue and cells in well-differentiated prostate carcinoma show great similarities in synthesis, storage, and release of prostasomes. However, this does not exclude the presence of other changes, for instance biochemical ones, in the prostasomes.

Adenocarcinoma↗

The effect of alpha-trinositol on cholera toxin-induced fluid accumulation in pig (Sus scrofa domesticus) jejunum in vivo.

The effect of alpha-trinositol (D-myo-inositol-1,2,6-trisphosphate) on cholera toxin-induced fluid accumulation (i.e., net fluid secretion) was studied in the pig jejunum in vivo. Cholera toxin caused a dose-dependent fluid accumulation in control experiments. Intravenous injection of alpha-trinositol produced a reduction of the response to cholera toxin with a significant maximal inhibition of 36%. However, in high concentrations of alpha-trinositol this inhibition was absent.

Animals↗

Effect of atropine on rat pancreatic secretory response to trypsin inhibitors and protein.

The effect of atropine on the exocrine pancreatic secretory response to intestinally infused trypsin inhibitor and protein in conscious rats was investigated. Bile and pancreatic juice were collected and continuously returned to the intestine throughout all experiments. Ovomucoid trypsin inhibitor (OMTI) was infused at 1, 3, 6, 12, and 30 mg/h id, simultaneously with intravenously infused atropine (100 micrograms X kg-1 X h-1) or 0.15 M NaCl. Casein was infused at 300 mg/h id with or without intravenous atropine. Atropine inhibited basal pancreatic protein and fluid secretion 65.7 and 24.7%, respectively. Atropine had no effect on incremental (above basal) pancreatic protein and fluid output during infusion of maximally effective doses of OMTI (12 and 30 mg/h) and increased the incremental responses to submaximal doses of OMTI and to casein. The results are consistent with the hypothesis that cholecystokinin mediates negative feedback regulation by luminal proteases and that cholinergic mechanisms are not directly involved in this regulatory mechanism.

Animals↗

Evaluation of human fetal urine as a source of amniotic fluid phospholipids.

The accumulation of surface active material in amniotic fluid during gestation is assumed to result from lung fluid secretion through the trachea. Some animal studies, however, have indicated that virtually all of the tracheal fluid is swallowed, whereas little if any enters the amniotic cavity. Following these observations fetal urine has been considered by some authors as an alternative source of amniotic fluid phospholipids. However, the phospholipid content of human fetal urine has not yet been determined. We have determined the L/S ratio and lamellar body concentration in human amniotic fluid and fetal urine obtained during 3 term cesarean deliveries. While both the L/S ratio and lamellar body particle concentration in the amniotic fluid samples were equivalent to values reported in term pregnancies, no measurable lecithin or sphingomyelin peaks were demonstrated in the urine samples and very few, if any, particles were counted. The lack of similarity between determinants of surface activity in human amniotic fluid and fetal urine does not support a major contribution of fetal urine to the phospholipid content of amniotic fluid.

Amniotic Fluid↗

A difference in stimulatory effects on pancreatic exocrine secretion between ursodeoxycholate and trypsin inhibitor in the rat.

We previously reported that intraduodenally infused ursodeoxycholate produced hypersecretion of pancreas in bicarbonate and fluid secretion in the rabbit (Digestive Diseases and Sciences, 28:942, 1983). Since trypsin inhibitor stimulates pancreatic secretion in the rat whose pancreatic exocrine secretion is regulated by a luminal feedback mechanism, in the present study we examined the stimulatory effect of ursodeoxycholate in comparison to Trasylol in unanesthetized rats with both the presence and the absence of returning bile-pancreatic juice. Under the condition in which bile-pancreatic juice were continuously returned to the intestine, the intraduodenally infused ursodeoxycholate produced significant increases in juice flow and bicarbonate and protein outputs, while Trasylol significantly increased protein output only. After an 8- to 10-hr period of bile-pancreatic juice diversion, Trasylol no longer affected pancreatic secretion, whereas ursodeoxycholate still stimulated the bicarbonate output significantly. Trypsin activities in the proximal half of the small intestine were not decreased by the infusion of UDCA. The mechanism of stimulatory effect of ursodeoxycholate on pancreatic secretion is independent of luminal feedback regulation and appears to differ from that of trypsin inhibitor.

Animals↗

The Salmonella enterica serotype typhimurium effector proteins SipA, SopA, SopB, SopD, and SopE2 act in concert to induce diarrhea in calves.

Salmonella enterica serotype Typhimurium requires a functional type III secretion system encoded by Salmonella pathogenicity island 1 (SPI1) to cause diarrhea. We investigated the role of genes encoding secreted target proteins of the SPI1-associated type III secretion system for enteropathogenicity in calves. Salmonella serotype Typhimurium strains having mutations in sptP, avrA, sspH1, or slrP induced fluid secretion in the bovine ligated ileal loop model at levels similar to that of the wild type. In contrast, mutations in sipA, sopA, sopB, sopD, or sopE2 significantly reduced fluid accumulation in bovine ligated ileal loops at 8 h postinfection. A strain carrying mutations in sipA, sopA, sopB, sopD, and sopE2 (sipA sopABDE2 mutant) caused the same level of fluid accumulation in bovine ligated ileal loops as a strain carrying a mutation in sipB, a SPI1 gene required for the translocation of effector proteins into host cells. A positive correlation was observed between the severity of histopathological lesions detected in the ileal mucosa and the levels of fluid accumulation induced by the different mutants. After oral infection of calves, the Salmonella serotype Typhimurium sipAsopABDE2 mutant caused only mild diarrhea and was more strongly attenuated than strains having only single mutations. These data demonstrate that SipA, SopA, SopB, SopD, and SopE2 are major virulence factors responsible for diarrhea during Salmonella serotype Typhimurium infection of calves.

Administration, Oral↗

Actions of some transmitters and their antagonists on salivary secretion in a tick.

Cholinomimetics (pilocarpine, carbachol, physostigmine, acetylcholine, acetyl-beta-methylcholine) and sympathomimetics (dopamine, epinephrine), when injected into the hemolymph, provoked salivary fluid secretion in the female ixodid tick Amblyomma hebraeum Koch. Atropine, but not tubocurarine or toxiferine, abolished pilocarpine-induced secretion without reducing the response to dopamine. Reserpine and guanethidine likewise selectively attenuated pilocarpine-induced secretion. Following extirpation of the synganglion, pilocarpine no longer provoked a secretory response whereas dopamine did. Thus, the salivary gland appears to be innervated directly by catecholaminergic rather than cholinergic secretory nerves. It is suggested that pilocarpine elicits salivation by interacting with muscarinic-type cholinergic receptors situated either on the cell bodies of the secretory nerves, or alternatively in the integrative or sensory pathway.

Acetylcholine↗