Intestinal Fluid Absorption Research Articles

Pathophysiology of Short Bowel Syndrome

The human small intestine is organized with a proximal-to-distal gradient of mucosal structure and nutrient processing capacity. However, certain nutrients undergo site-specific digestion and absorption, such as iron and folate in the duodenum/jejunum vs vitamin B12 and bile salts in the ileum. Intestinal resection can result in short bowel syndrome (SBS) due to reduction of total and/or site-specific nutrient processing areas. Depending on the segment(s) of intestine resected, malabsorption can be nutrient specific (eg, vitamin B12 or fat) or sweeping, with deficiencies in energy, protein, and various micronutrients. Jejunal resections are generally better tolerated than ileal resections because of greater postresection adaptive capacity than that of the jejunum. Following intestinal resection, energy scavenging and fluid absorption become particularly important in the colon owing to loss of digestive and absorptive surface area in the resection portion. Resection-induced alterations in enteroendocrine cell abundance can further disrupt intestinal function. For example, patients with end jejunostomy have depressed circulating peptide YY and glucagon-like peptide 2 concentrations, which likely contribute to the rapid intestinal transit and blunted intestinal adaptation observed in this population. SBS-associated pathophysiology often extends beyond the gastrointestinal tract, with hepatobiliary disease, metabolic bone disease, D-lactic acidosis, and kidney stone formation being chronic complications. Clinical management of SBS must be individualized to account for the specific nutrient processing deficit within the remnant bowel and to mitigate potential complications, both inside and outside the gastrointestinal tract.

The distinct roles of anion transporters Slc26a3 (DRA) and Slc26a6 (PAT-1) in fluid and electrolyte absorption in the murine small intestine

The mixing of gastric and pancreatic juice subjects the jejunum to unique ionic conditions with high luminal CO2 tension and HCO3 − concentration. We investigated the role of the small intestinal apical anion exchangers PAT-1 (Slc26a6) and DRA (Slc26a3) in basal and CO2/HCO3 −-stimulated jejunal fluid absorption. Single pass perfusion of jejunal segments was performed in anaesthetised wild type (WT) as well as in mice deficient in DRA, PAT-1, Na+/H+ exchanger 3 (NHE3) or NHE2, and in carbonic anhydrase II (CAII). Unbuffered saline (pH 7.4) perfusion of WT jejunum resulted in fluid absorption and acidification of the effluent. DRA-deficient jejunum absorbed less fluid than WT, and acidified the effluent more strongly, consistent with its action as a Cl−/HCO3 − exchanger. PAT-1-deficient jejunum also absorbed less fluid but resulted in less effluent acidification. Switching the luminal solution to a 5 % CO2/HCO3 − buffered solution (pH 7.4), resulted in a decrease in jejunal enterocyte pHi in all genotypes, an increase in luminal surface pH and a strong increase in fluid absorption in a PAT-1- and NHE3- but not DRA-, CAII, or NHE2-dependent fashion. Even in the absence of luminal Cl−, luminal CO2/HCO3 − augmented fluid absorption in WT, CAII, NHE2- or DRA-deficient, but not in PAT-1- or NHE3-deficient mice, indicating the likelihood that PAT-1 serves to import HCO3 − and NHE3 serves to import Na+ under these circumstances. The results suggest that PAT-1 plays an important role in jejunal Na+HCO3 – reabsorption, while DRA absorbs Cl− and exports HCO3 − in a partly CAII-dependent fashion. Both PAT-1 and DRA significantly contribute to intestinal fluid absorption and enterocyte acid/base balance but are activated by different ion gradients.Electronic supplementary materialThe online version of this article (doi:10.1007/s00424-013-1381-2) contains supplementary material, which is available to authorized users.

Fructose–Maltodextrin Ratio Governs Exogenous and Other CHO Oxidation and Performance

Fructose coingested with glucose in carbohydrate (CHO) drinks increases exogenous-CHO oxidation, gut comfort, and physical performance. This study aimed to determine the effect of different fructose-maltodextrin-glucose ratios on CHO oxidation and fluid absorption while controlling for osmolality and caloricity. In a crossover design, 12 male cyclists rode 2 h at 57% peak power then performed 10 sprints while ingesting artificially sweetened water or three equiosmotic 11.25% CHO-salt drinks at 200 mL·15 min, comprising weighed fructose and maltodextrin-glucose in ratios of 0.5:1 (0.5 ratio), 0.8:1 (0.8 ratio), and 1.25:1 (1.25 ratio). Fluid absorption was traced with D2O, whereas C-fructose and C-maltodextrin-glucose permitted fructose and glucose oxidation rate evaluation. The mean exogenous-fructose and exogenous-glucose oxidation rates were 0.27, 0.39, and 0.46 g·min and 0.65, 0.71, and 0.58 g·min in 0.5, 0.8, and 1.25 ratio drinks, representing mean oxidation efficiencies of 54%, 59%, and 55% and 65%, 85%, and 86% for fructose and glucose, respectively. With the 0.8 ratio drink, total exogenous-CHO oxidation rate was 18% (90% confidence interval, ±5%) and 5.2% (±4.6%) higher relative to 0.5 and 1.25 ratios, respectively, whereas respective differences in total exogenous-CHO oxidation efficiency were 17% (±5%) and 5.3% (±4.8%), associated with 8.6% and 7.8% (±4.2%) higher fructose oxidation efficiency. The effects of CHO ratio on water absorption were inconclusive. Mean sprint power with the 0.8 ratio drink was moderately higher than that with the 0.5 ratio (2.9%; 99% confidence interval, ±2.8%) and 1.25 ratio (3.1%; ±2.7%) drinks, with total- and endogenous-CHO oxidation rate, abdominal cramps, and drink sweetness qualifying as explanatory mechanisms. Enhanced high-intensity endurance performance with a 0.8 ratio fructose-maltodextrin-glucose drink is characterized by higher exogenous-CHO oxidation efficiency and reduced endogenous-CHO oxidation. The gut-hepatic or other physiological site responsible requires further research.

Effects of faba bean and faba bean hulls on expression of selected genes in the small intestine of piglets

In a small intestinal segment perfusion (SISP) study in pigs, effects were studied of intestinal perfusion of ground faba beans (Vicia faba), faba bean hulls, or saline on intestinal net fluid absorption in intestinal segments either challenged or not with an enterotoxigenic Escherichia coli (ETEC). After an 8-h perfusion, piglets were euthanized and small intestinal mucosa samples were collected for analysis of expression level of a selected set of genes (APOC3, TIMP1, AQP8, MMP1, MUC13, and PAP) using real-time quantitative PCR (qPCR). Perfusion with ground faba beans and faba bean hulls and challenge with ETEC affected (P < 0.05) expression of several of genes in the intestinal mucosa. Expression of APOC3, TIMP1, AQP8, MMP1, and PAP was correlated with net fluid absorption in the small intestine of pigs.

Covering the Cover

Covering the Cover

Optimization of a small intestinal segment perfusion model for heat-stable enterotoxin A induced secretion in pigs

Enterotoxigenic Escherichia coli (ETEC) are a major cause of infectious diarrhea both in human and pigs. After ingestion of contaminated food or water, ETEC bacteria colonize the small intestine where they produce heat-labile (LT) and/or heat-stable (ST) enterotoxins, which induce watery diarrhea. We investigated the possibility of eliciting STa-induced secretion in jejunal segments of anesthetized pigs using a small-intestinal segment perfusion (SISP) model. Five consecutive mid-jejunal segments of anaesthetized piglets were perfused for 6h with different concentrations of STa in a physiologic salt solution. Changes in intestinal net fluid absorption were measured. From the results we could conclude that the STa response was dose-dependent and that continuous perfusion with 50nM of STa or more was required to reduce net absorption. This concentration was sufficient to reduce net absorption compared to control segments in 12 out of 14 piglets. STa-induced responses however showed relative high variation between different jejunal segments of one pig, similar to the inter-segment variation seen in control animals where segments were perfused with physiologic salt solution. These results indicate that more optimization is required before this model could be used to test compounds that could interfere with the STa-induced fluid secretion.

Teduglutide, a novel glucagon-like peptide 2 analog, in the treatment of patients with short bowel syndrome.

Short bowel syndrome results from surgical resection, congenital defect or disease-associated loss of absorption. Parenteral support (PS) is lifesaving in patients with short bowel syndrome and intestinal failure who are unable to compensate for their malabsorption by metabolic or pharmacologic adaptation. Together, the symptoms of short bowel syndrome and the inconvenience and complications in relation to PS (e.g. catheter-related blood steam infections, central thrombosis and intestinal failure associated liver disease) may impair the quality of life of patients. The aim of treatment is to maximize intestinal absorption, minimize the inconvenience of diarrhea, and avoid, reduce or eliminate the need for PS to achieve the best possible quality of life for the patient. Conventional treatments include dietary manipulations, oral rehydration solutions, and antidiarrheal and antisecretory treatments. However, the evidence base for these interventions is limited and treatments that improve the structural and functional integrity of the remaining intestine are needed. Teduglutide, an analog of glucagon-like peptide 2, improves intestinal rehabilitation by promoting mucosal growth and possibly by restoring gastric emptying and secretion, thereby reducing intestinal losses and promoting intestinal absorption. In a 3-week, phase II balance study, teduglutide reduced diarrhea by around 700 g/day and fecal energy losses by around 0.8 MJ/day. In two randomized, placebo-controlled, 24-week, phase III studies, similar findings were obtained when evaluating the fluid composite effect, which is the sum of the beneficial effects of teduglutide - reduction in the need for PS, increase in urine production and reduction in oral fluid intake. The fluid composite effect reflects the increase in intestinal fluid absorption (and the concomitant reduction in diarrhea) and may be used in studies in which metabolic balance assessments are not performed. In studies of up to 24 weeks' duration, teduglutide appears to be safe and well tolerated. Treatment with teduglutide was associated with enhancement or restoration of the structural and functional integrity of the remaining intestine with significant intestinotrophic and proabsorptive effects, facilitating a reduction in diarrhea and an equivalent reduction in the need for PS in patients with short bowel syndrome and intestinal failure.

Intestinal fluid absorption in anadromous salmonids: importance of tight junctions and aquaporins

The anadromous salmonid life cycle includes both fresh water (FW) and seawater (SW) stages. The parr-smolt transformation (smoltification) pre-adapt the fish to SW while still in FW. The osmoregulatory organs change their mode of action from a role of preventing water inflow in FW, to absorb ions to replace water lost by osmosis in SW. During smoltification, the drinking rate increases, in the intestine the ion and fluid transport increases and is further elevated after SW entry. In SW, the intestine absorbs ions to create an inwardly directed water flow which is accomplished by increased Na+, K+-ATPase (NKA) activity in the basolateral membrane, driving ion absorption via ion channels and/or co-transporters. This review will aim at discussing the expression patterns of the ion transporting proteins involved in intestinal fluid absorption in the FW stage, during smoltification and after SW entry. Of equal importance for intestinal fluid absorption as the active absorption of ions is the permeability of the epithelium to ions and water. During the smoltification the increase in NKA activity and water uptake in SW is accompanied by decreased paracellular permeability suggesting a redirection of the fluid movement from a paracellular route in FW, to a transcellular route in SW. Increased transcellular fluid absorption could be achieved by incorporation of aquaporins (AQPs) into the enterocyte membranes and/or by a change in fatty acid profile of the enterocyte lipid bilayer. An increased incorporation of unsaturated fatty acids into the membrane phospholipids will increase water permeability by enhancing the fluidity of the membrane. A second aim of the present review is therefore to discuss the presence and regulation of expression of AQPs in the enterocyte membrane as well as to discuss the profile of fatty acids present in the membrane phospholipids during different stages of the salmonid lifecycle.

Antidiarrheal efficacy of a quinazolin CFTR inhibitor on human intestinal epithelial cell and in mouse model of cholera

Objective:This study aimed to evaluate the antidiarrheal efficacy and pharmacological properties of ethyl 2-(4-oxo-3-o-tolyl-3,4-dihydroquinazolin-2-ylthio)acetate (DQA) as an inhibitor of cystic fibrosis transmembrane conductance regulator protein (CFTR) both in vitro and in vivo.Materials and Methods:The effects of DQA on CFTR function and cell viability were investigated in Fisher rat thyroid (FRT) cells expressing human CFTR and human intestinal epithelial T84 cells by short-circuit current measurements and MTT assays, respectively. In vivo antidiarrheal efficacy of DQA was evaluated in a closed loop model of cholera in mice.Results:In permeabilized FRT cells, apical chloride current induced by CFTR agonists (10 μM forskolin, 100 μM CPT-cAMP, and 20 μM apigenin) was inhibited by DQA with IC50 ~ 20 μM and complete inhibition at 200 μM. The inhibitory effect was reversible and not associated with cytotoxicity to FRT cells (5–500 μM DQA for 24 h). Likewise, DQA effectively inhibited both forskolin and cholera toxin-induced transepithelial chloride secretion in T84 cells. In mice, intraluminal injection of 100 μM DQA reduced cholera toxin (1 μg/closed loop)-induced intestinal fluid secretion by 85% without affecting intestinal fluid absorption.Conclusions:DQA represents a new class of small molecule CFTR inhibitor with potential application in treatment of cholera.

Osmoregulation and epithelial water transport: lessons from the intestine of marine teleost fish

For teleost fish living in seawater, drinking the surrounding medium is necessary to avoid dehydration. This is a key component of their osmoregulatory strategy presenting the challenge of excreting excess salts while achieving a net retention of water. The intestine has an established role in osmoregulation, and its ability to effectively absorb fluid is crucial to compensating for water losses to the hyperosmotic environment. Despite this, the potential for the teleost intestine to serve as a comparative model for detailed, integrative experimental studies on epithelial water transport has so far gone largely untapped. The following review aims to present an assessment of the teleost intestine as a fluid-transporting epithelium. Beginning with a brief overview of marine teleost osmoregulation, emphasis shifts to the processing of ingested seawater by the gastrointestinal tract and the characteristics of intestinal ion and fluid transport. Particular attention is given to acid-base transfers by the intestine, specifically bicarbonate secretion, which creates the distinctly alkaline gut fluids responsible for the formation of solid calcium carbonate precipitates. The respective contributions of these unique features to intestinal fluid absorption, alongside other recognised ion transport processes, are then subsequently considered within the wider context of the classic physiological problem of epithelial water transport.

Intestinal anion exchange in marine teleosts is involved in osmoregulation and contributes to the oceanic inorganic carbon cycle

Marine teleost fish osmoregulation involves seawater ingestion and intestinal fluid absorption. Solute coupled fluid absorption by the marine teleost fish intestine has long been believed to be the product of Na(+) and Cl(-) absorption via the Na(+) :K(+) :2Cl(-) co-transporter (NKCC2). However, the past decade has revealed that intestinal anion exchange contributes significantly to Cl(-) absorption, in exchange for HCO(3) (-) secretion, and that this process is important for intestinal water absorption. In addition to contributing to solute coupled water absorption intestinal anion exchange results in luminal precipitation of CaCO(3) which acts to reduce luminal osmotic pressure and thus assist water absorption. Most recently, activity of apical H(+) -pumps, especially in distal segments of the intestine have been suggested to not only promote anion exchange, but also to reduce luminal osmotic pressure by preventing excess HCO(3)(-) concentrations from accumulating in intestinal fluids, thereby aiding water absorption. The present review summarizes and synthesizes the most recent advances in our view of marine teleosts osmoregulation, including our emerging understanding of epithelial transport of acid-base equivalents in the intestine, the consequences for whole organism acid-base balance and finally the impact of piscine CaCO(3) formation on the global oceanic carbon cycle.

Randomised placebo-controlled trial of teduglutide in reducing parenteral nutrition and/or intravenous fluid requirements in patients with short bowel syndrome

Background and aimsTeduglutide, a GLP-2 analogue, may restore intestinal structural and functional integrity by promoting repair and growth of the mucosa and reducing gastric emptying and secretion, thereby increasing fluid...

Intestinal Barrier Physiology

Abstract Enterotoxigenic Escherichia coli (ETEC) are a major cause of infectious diarrhea both in human and pigs. After ingestion of contaminated food or water, ETEC bacteria colonize the small intestine where they produce heat-labile (LT) and/or heat-stable (ST) enterotoxins, which induce watery diarrhea. We investigated the possibility of eliciting STa-induced secretion in jejunal segments of anesthetized pigs using a small-intestinal segment perfusion (SISP) model. Five consecutive mid-jejunal segments of anaesthetized piglets were perfused for 6 h with different concentrations of STa in a physiologic salt solution. Changes in intestinal net fluid absorption were measured. From the results we could conclude that the STa response was dose-dependent and that continuous perfusion with 50 nM of STa or more was required to reduce net absorption. This concentration was sufficient to reduce net absorption compared to control segments in 12 out of 14 piglets. STa-induced responses however showed relative high variation between different jejunal segments of one pig, similar to the inter-segment variation seen in control animals where segments were perfused with physiologic salt solution. These results indicate that more optimization is required before this model could be used to test compounds that could interfere with the STa-induced fluid secretion.

Intestinal Fluid and Glucose Transport in Wistar Rats following Chronic Consumption of Fresh or Oxidised Palm Oil Diet

Chronic ingestion of thermoxidized palm oil causes functional derangement of various tissues. This study was therefore carried out to determine the effect of chronic ingestion of thermoxidized and fresh palm oil diets on intestinal fluid and glucose absorption in rats using the everted sac technique. Thirty Wistar rats were divided into three groups of 10 rats per group. The first group was the control and was fed on normal rat chow while the second (FPO) and third groups (TPO) were fed diet containing either fresh or thermoxidized palm oil (15% wt/wt) for 14 weeks. Villus height and crypt depth were measured. The gut fluid uptake and gut glucose uptake were significantly (P < .001) lower in the TPO group than those in the FPO and control groups, respectively. The villus height in the TPO was significantly (P < .01) lower than that in FPO and control. The villus depth in TPO was significantly (P < .05) higher than that in FPO and control groups, respectively. These results suggest that ingestion of thermoxidized palm oil and not fresh palm oil may lead to distortion in villus morphology with a concomitant malabsorption of fluid and glucose in rats due to its harmful free radicals.

NANETS Consensus Guidelines for the Diagnosis of Neuroendocrine Tumor

Neuroendocrine tumors (NETs) are rare, slow-growing neoplasms characterized by their ability to store and secrete different peptides and neuroamines. 1 Some of these substances cause specific clinical syndromes, 2 whereas other may have elevated plasma or urine levels that are not associated with specific syndromes or symptom complexes Unfortunately, there is no Bideal neuroendocrine tumor marker,[ 3 but according to the presentation, the sensitivity and specificity of each marker vary, and it is generally possible to choose those of greatest value for each clinical syndrome. The biochemical markers are those hormones or amines secreted by the neuroendocrine cells from which these tumors are derived. Some of these are not specific to any tumor, but are produced and secreted by most NETs, whereas other biochemical markers are more specific to the type of tumor and where their quantification can lead to the suspicion or confirmation of the presence of such a tumor. The annual incidence of NETs has risen to 40 to 50 cases per million, perhaps because of better diagnosis and the availability of highly specific and sensitive ways to measure these tumors’ products, improved immunohistochemistry, and enhanced techniques for tumor detection. Thus, the perceived increase in incidence may not be a real change in the incidence of the disease. There are a number of impediments to the diagnosis of these tumors. They are rare, comprising less than 2% of gastrointestinal (GI) malignancies, and are therefore not high on the list of causes of specific symptom complexes. Symptoms themselves are often nonspecific and do not lend themselves readily to identifying the specific underlying tumor. In addition, the manifestations are protean and mimic a variety of disorders. Tumors may be found incidentally on laparoscopy for abdominal pain or during the surgical removal of an appendix or even during a computerized tomographic scan of the abdomen for unexplained symptoms. Lung carcinoids may present with hemoptysis or asthma-like symptoms, and midgut carcinoids may be confused with irritable bowel syndrome (IBS). The natural history of this disease is invariably attended by a long history of vague abdominal symptoms, a series of visits to a primary care practitioner, and referral to a gastroenterologist, often with a misdiagnosis of IBS. These symptoms persist with a median latency to correct diagnosis of 9.2 years by which time the tumor has metastasized, causing symptoms such as flushing and diarrhea and progressing on its slow but relentless course until the patient dies. Clearly, a greater index of suspicion and a carcinoid tumor profile screen are warranted for all patients presenting with Btraditional IBS symptoms.[ Midgut carcinoids are associated with mesenteric fibrosis, which can compress mesenteric vessels and cause bowel ischemia and malabsorption, which may be found in the absence of an abdominal mass. The diagnosis of metastases to the liver is generally more obvious but often still takes place only after a delay of many years. Even then, an incorrect diagnosis is not uncommon. Unless biopsy material is examined for the secretory peptides chromogranin, synaptophysin, or neuron-specific enolase (NSE), tumors may be labeled erroneously as adenocarcinoma, with a negative impact on physician’s attitudes regarding management and underestimation of prospects for survival. 4 The common symptomatic manifestations of patients with carcinoid tumors are illustrated in Tables 1 and 2. Flushing

Effect of pea and faba bean fractions on net fluid absorption in ETEC-infected small intestinal segments of weaned piglets

After weaning piglets frequently have diarrhoea associated with an enterotoxigenic Escherichia coli (ETEC) infection. Alternative plant protein sources such as peas, faba beans and lupins may contribute in preventing gastrointestinal problems. In the small intestinal segment perfusion model, the anti-diarrhoea effect of pea and faba bean fractions on net absorption of fluid was investigated. Segments with a cranial inflow tube and a caudal outflow tube were fitted in the small intestine of 5-week old anaesthetized piglets. The odd numbered segments were infused with ETEC and the even segments with PBS, whereupon the segments were perfused over 8 h. Pea meal, pea hulls, protein-enriched pea meal and starch-enriched pea meal were perfused in 4 piglets in 4 pairs of segments (an uninfected and an adjacent ETEC-infected) with saline as a control in another pair of segments. Faba bean meal, faba bean hulls, protein-enriched faba bean meal and starch-enriched faba bean meal were perfused in another 4 piglets. Net fluid absorption was calculated from the difference between the volumes of inflow and outflow divided by the surface area. ETEC induced loss of net intestinal fluid absorption was shown to be decreased in the segments perfused with pea hulls and faba bean hulls, while there was no effect of the other pea or faba bean products. Although hulls of pea and faba bean did not completely eliminate the decreased fluid absorption after ETEC challenge, these results indicate that hulls of pea and faba bean may promote net fluid absorption in piglets during post weaning diarrhoea.

Gene ablation for PEPT1 in mice abolishes the effects of dipeptides on small intestinal fluid absorption, short-circuit current, and intracellular pH

PEPT1 function in mouse intestine has not been assessed by means of electrophysiology and methods to assess its role in intracellular pH and fluid homeostasis. Therefore, the effects of the dipeptide glycilsarcosin (Gly-Sar) on jejunal fluid absorption and villous enterocyte intracellular pH (pH(i)) in vivo, as well as on enterocyte[(14)C]Gly-Sar uptake, short-circuit current (I(sc)) response, and enterocyte pH(i) in vitro were determined in wild-type and PEPT1-deficient mice and in mice lacking PEPT1. Immunohistochemistry for PEPT1 failed to detect any protein in enterocyte apical membranes in Slc15a1(-/-) animals. Saturable Gly-Sar uptake in Slc15a1(-/-) everted sac preparations was no longer detectable. Similarly, Gly-Sar-induced jejunal I(sc) response in vitro was abolished. The dipeptide-induced increase in fluid absorption in vivo was also abolished in animals lacking PEPT1. Since PEPT1 acts as an acid loader in enterocytes, enterocyte pH(i) was measured in vivo by two-photon microscopy in SNARF-4-loaded villous enterocytes of exteriorized jejuni in anesthetized mice, as well as in BCECF-loaded enterocytes of microdissected jejunal villi. Gly-Sar-induced pH(i) decrease was no longer observed in the absence of PEPT1. A reversal of the proton gradient across the luminal membrane did not significantly diminish Gly-Sar-induced I(sc) response, whereas a depolarization of the apical membrane potential by high K(+) or via Na(+)-K(+)-ATPase inhibition strongly diminished Gly-Sar-induced I(sc) responses. This study demonstrates for the first time that proton-coupled electrogenic dipeptide uptake in the native small intestine, mediated by PEPT1, relies on the negative apical membrane potential as the major driving force and contributes significantly to intestinal fluid transport.

A Plant-Derived Hydrolysable Tannin Inhibits CFTR Chloride Channel: A Potential Treatment of Diarrhea

The present study examined the effects and mechanisms of actions of penta-m-digalloyl-glucose (PDG), a hydrolysable tannin extracted from Chinese gallnut, on cystic fibrosis transmembrane conductance regulator protein (CFTR). Fisher rat thyroid cells stably expressing human CFTR (FRT cells) and human intestinal T84 cells were used as cell models to investigate the effects of PDG on chloride secretion using short-circuit current analysis. The mechanisms by which PDG affected chloride secretion were also examined. Finally, in vivo antidiarrheal efficacy and effects of PDG on intestinal fluid absorption were evaluated in mouse closed-loop models. In FRT cells, apical chloride current induced by forskolin, CPT-cAMP and apigenin were reversibly inhibited by PDG (IC50 approximately 10microM) without effects on intracellular cAMP content and cell viability. Similarly, in T84 cells, PDG effectively inhibited chloride secretion induced by forskolin and cholera toxin. However, it had no effect on calcium-induced chloride secretion. In mice, a single intraluminal injection of PDG (0.6 mg/kg) reduced cholera toxin-induced intestinal fluid secretion by 75% with no effect on intestinal fluid absorption. PDG represents a new class of CFTR inhibitors. Further development of this class of compounds may provide a new therapeutic intervention for diarrhea.

Increasing weaning age of piglets from 4 to 7 weeks reduces stress, increases post-weaning feed intake but does not improve intestinal functionality

This study tested the hypothesis that late weaning and the availability of creep feed during the suckling period compared with early weaning, improves feed intake, decreases stress and improves the integrity of the intestinal tract. In this study with 160 piglets of 16 litters, late weaning at 7 weeks of age was compared with early weaning at 4 weeks, with or without creep feeding during the suckling period, on post-weaning feed intake, plasma cortisol (as an indicator of stress) and plasma intestinal fatty acid binding protein (I-FABP; a marker for mild intestinal injury) concentrations, intestinal morphology, intestinal (macro)molecular permeability and intestinal fluid absorption as indicators of small intestinal integrity. Post-weaning feed intake was similar in piglets weaned at 4 weeks and offered creep feed or not, but higher (P < 0.001) in piglets weaned at 7 weeks with a higher (P < 0.05) intake for piglets offered creep feed compared with piglets from whom creep feed was witheld. Plasma cortisol response at the day of weaning was lower in piglets weaned at 7 weeks compared with piglets weaned at 4 weeks, and creep feed did not affect cortisol concentration. Plasma I-FABP concentration was not affected by the age of weaning and creep feeding. Intestinal (macro)molecular permeability was not affected by the age of weaning and creep feeding. Both in uninfected and enterotoxigenic Escherichia coli-infected small intestinal segments net fluid absorption was not affected by the age of weaning or creep feeding. Creep feeding, but not the age of weaning, resulted in higher villi and increased crypt depth. In conclusion, weaning at 7 weeks of age in combination with creep feeding improves post-weaning feed intake and reduces weaning stress but does not improve functional characteristics of the small intestinal mucosa.

Lysophosphatidic Acid Stimulates the Intestinal Brush Border Na +/H + Exchanger 3 and Fluid Absorption via LPA 5 and NHERF2

Diarrhea results from reduced net fluid and salt absorption caused by an imbalance in intestinal absorption and secretion. The bulk of sodium and water absorption in the intestine is mediated by Na(+)/H(+) exchanger 3 (NHE3), located in the luminal membrane of enterocytes. We investigated the effect of lysophosphatidic acid (LPA) on Na(+)/H(+) exchanger activity and Na(+)-dependent fluid absorption in the intestine. We analyzed the effects of LPA on fluid absorption in intestines of wild-type mice and mice deficient in Na(+)/H(+) exchanger regulatory factor 2 (NHERF2; Nherf2(-/-)) or LPA(2) (Lpa(2)(-/-)). Roles of LPA(5) and NHERF2 were determined by analysis of heterologous expression. Under basal conditions, LPA increased fluid absorption in an NHE3-dependent manner and restored the net fluid loss in a mouse model of acute diarrhea. Expression of the LPA receptor LPA(5) was necessary for LPA-induced stimulation of NHE3 activity in colonic epithelial cells. Stimulation of NHE3 by the LPA-LPA(5) signaling required coexpression of NHERF2, which interacted with LPA(5). LPA-mediated intestinal fluid absorption was impaired in Nherf2(-/-) mice, demonstrating the requirement for NHERF2 in LPA(5) activity. However, fluid absorption was unaltered in Lpa(2)(-/-) mice. LPA stimulated NHE3 and fluid absorption in part by increasing NHE3 protein abundance at the brush border membrane of intestinal epithelial cells. LPA is a potent stimulant of NHE3 and fluid absorption in the intestine, signaling through LPA(5). Regulation by LPA(5) depends on its interaction with NHERF2. LPA might be useful in the treatment of certain diarrheal diseases.