Paracellular absorption in laboratory mice: Molecule size-dependent but low capacity.

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular processes. The capacity for paracellular absorption seems lower in nonfliers than in fliers, although that conclusion rests largely on a comparison of relatively larger nonflying mammals (>155g) and relatively smaller flying birds (<155g). We report on paracellular absorption in laboratory mice, the smallest nonflying mammal species studied to date. Using a standard pharmacokinetic technique, we measured the extent of absorption (fractional absorption=f) of inert carbohydrate probes: L-arabinose (M(r)=150.13Da) and cellobiose (342.3) that are absorbed exclusively by the paracellular route, and 3-O-methyl D-glucose (3OMD-glucose) (M(r)=194) absorbed both paracellularly and transcellularly. f was measured accurately in urine collection trials of 5-10h duration. Absorption of 3OMD-glucose by mice was essentially complete (f=0.95±0.07) and much higher than that for L-arabinose (f=0.21±0.02), indicating that in mice, like other nonflying mammals, >80% of glucose is absorbed by mediated process(es) rather than the passive, paracellular route. As in all other vertebrates, absorption of cellobiose (f=0.13±0.02) was even lower than that for L-arabinose, suggesting an equivalent molecular size cut-off for flying and nonflying animals and thus a comparable effective TJ aperture. An important ecological implication is that smaller water-soluble plant secondary metabolites that have been shown to be absorbed by the paracellular path in cell culture, such as phenolics and alkaloids, might be absorbed in substantial amounts by bats and small birds relative to nonflying mammals such as mice.

Intestinal function is impaired in patients with Chronic Obstructive Pulmonary Disease.

BACKGROUND & AIMS: Gastrointestinal symptoms are prevalent extrapulmonary systemic manifestations of Chronic Obstructive Pulmonary Disease (COPD), but have been rarely studied. We dissected the perturbations in intestinal function in human patients with COPD using comprehensive metabolic and physiological approaches. METHODS: In this observational study, small intestinal membrane integrity and active carrier-mediated glucose transport were quantified by sugar permeability test in 21 clinically stable patients with moderate to severe COPD (mean FEV1, 41.2 (3.2) % of predicted) and 16 healthy control subjects. Protein digestion and absorption was analyzed using stable tracer kinetic methods. Plasma acetate, propionate, and butyrate concentrations were measured as markers of intestinal microbial metabolism. RESULTS: Compared with healthy controls, non carrier-mediated permeability was higher (0.062 (95% CI [0.046, 0.078]) vs. 0.037 (95% CI [0.029, 0.045]), P = 0.009) and active glucose transport lower in COPD (31.4 (95% CI [23.4, 39.4])% vs. 48.0 (95% CI [37.8, 58.3])%, P = 0.010). Protein digestion and absorption was lower in COPD (0.647 (95% CI [0.588, 0.705]) vs. 0.823 (95% CI [0.737, 0.909]), P = 0007), and impairment greater in patients with dyspnea (P = 0.038), exacerbations in preceding year (P = 0.052), and reduced transcutaneous oxygen saturation (P = 0.051), and was associated with reduced physical activity score (P = 0.016) and lower quality of life (P = 0.0007). Plasma acetate concentration was reduced in COPD (41.54 (95% CI [35.17, 47.91]) vs. 80.44 (95% CI [54.59, 106.30]) µmol/L, P = 0.001) suggesting perturbed intestinal microbial metabolism. CONCLUSIONS: We conclude that intestinal dysfunction is present in COPD, worsens with increasing disease severity, and is associated with reduced quality of life.

A blood test for intestinal permeability and function: a new tool for the diagnosis of chronic intestinal disease in dogs.

We demonstrate that rhamnose, 3-O-methyl-D-glucose, D-xylose and lactulose may be quantified accurately in blood by HPLC and pulsed amperometric detection, thus enabling studies of intestinal permeability and function to be carried out using plasma samples. Prior to HPLC, the endogenous glucose was enzymatically modified to gluconic acid and the protein precipitated. The precision of the quantification of the sugars in plasma (CV: 2.2-5.7%; 8.7-10.6% at very low concentrations) compared well with the quantification in urine. The results for groups of 8 dogs with small intestinal bacterial overgrowth and 12 dogs with inflammatory bowel disease were shown to be significantly different from a group of 20 normal control dogs (P < 0.001), demonstrating the test's value as a diagnostic tool. The normal ranges in blood 2 h post oral administration were determined to be 0.05-0.17 for the lactulose/rhamnose ratio and 0.45-0.65 for the xylose/3-O-methylglucose ratio. This method may be employed advantageously when the collection of urine in intestinal permeability and function tests is difficult.

3-0 methylglucose uptake as a marker of nutrient absorption and bowel length in pediatric patients.

BACKGROUND: Inert carbohydrate probes are commonly used to assess intestinal permeability; we have previously shown that the actively transported moiety 3-0 methylglucose (3-0 MG) is a useful marker of intestinal surface area and nutrient absorption in animal models of short bowel syndrome (SBS). This study examines the correlation of 3-0 MG absorption with nutrient absorption, bowel length, and the tolerance of enteral feeds in pediatric patients. METHODS: Fifteen children (1 month to 15 years in age) were studied after intestinal surgery. All had a stoma, 2 were > 1 year of age, the remainder had surgical intervention as a neonate or within the first month of life. Eight had SBS (50% expected bowel length for age). Bowel length was measured intraoperatively. Nutrient absorption was quantified with a 48-hour bowel study, measuring fat, protein, and carbohydrate output directly. 3-0 MG absorption and intestinal permeability were quantified using a solution containing 30 mg/mL 3-0 MG, 20 mg/mL mannitol and 30 mg/mL lactulose (osmolarity 352, given at 1 mL/kg via feeding tube). Subsequent urine production was collected for 8 hours, and probe recovery measured using HPLC. RESULTS: 3-0 MG absorption was significantly correlated with nutrient absorption. The correlation with protein absorption was r2 = .59, fat r2 = .62 and carbohydrate r2 = .56. The correlation between 3-0 MG absorption and bowel length was r2 = .58. 3-0 MG absorption was significantly lower in SBS patients vs patients with normal bowel length (15.8 +/- 6.7 vs 30.5 +/- 10.2%). 3-0 MG absorption also correlated with the ability to tolerate enteral feeds (r2 = .38; p < .03 for all comparisons). CONCLUSIONS: 3-0 MG may be a useful marker of nutrient absorption and bowel length in pediatric patients with short bowel syndrome. The simplicity and reproducibility of the method make it an attractive option for following patient outcomes. Further studies are suggested to determine the utility of these markers in directing the clinical management of patients.

In vivo measurement of intestinal absorption using 3-0 methylglucose in short bowel syndrome.

PURPOSE: The management of patients with short bowel syndrome is complicated by the paucity of methods to assess in vivo the absorptive capacity of the remaining bowel. The purpose of this experiment was to assess the feasibility of using urinary recovery of 3-0 methylglucose (3-0 MG) as a quantitative measure of carbohydrate absorptive capacity, comparing it with in vivo absorption and in vitro glucose transport studies. METHODS: Male Sprague Dawley rats underwent either a 90% proximal small bowel resection or sham resection (n = 8 in each group). Animals were pair fed, weighed, and followed up for 14 days. A 3-day balance study was done, measuring feed intake and fecal output for percentages of fat and energy absorption. Animals were gavaged with 3-0 MG/Mannitol solution, and 4-hour urinary recovery of sugars was assessed using high-performance liquid chromatography (HPLC). On different days these studies were repeated with increasing amounts of added normal glucose (1 mol/L, 1.25 mol/L, and 1.5 mol/L) in the gavage solution given to compete for 3-0 MG transport, and thus increase the "sensitivity" of the test. Animals were then killed, and sections of intestine taken for in vitro assessment of glucose transport using radiolabeled 3-0 MG in Ussing chambers. RESULTS: Total energy, carbohydrate, and fat absorption all were reduced significantly in the resected animals, as was 3-0 MG urinary recovery (62.9 +/- 10.5%) in controls versus (35.8 +/- 17.5%) in resected animals (P <.05). 3-0 MG urinary recovery correlated well with dietary carbohydrate absorption (r = 0.74), and with Ussing chamber measures of glucose flux (r = 0.97). Adding exogenous glucose to the test solution to "compete" for 3-0 MG transport sites did not improve sensitivity. CONCLUSIONS: These results show that 3-0 MG is useful in measuring nutrient absorption capacity in rats after massive small bowel resection. Further studies to validate these methods in human patients with short bowel syndrome are suggested.

Introduction of enteral feeding in neonates on extracorporeal membrane oxygenation after evaluation of intestinal permeability changes.

BACKGROUND/PURPOSE: Neonates meeting criteria for extracorporeal membrane oxygenation (ECMO) often suffer from variable periods of hypoxia. During ECMO, starvation of the gut is common practice in many centres as splanchnic ischemia results in loss of intestinal integrity, which in turn predisposes for bacterial translocation and sepsis and eventually necrotizing enterocolitis (NEC) and multiorgan failure. However, minimal enteral feeding is thought to be of benefit in the critically ill. Data on intestinal integrity in newborns on ECMO and the effects of enteral nutrition are not available. This study prospectively evaluates the changes in small intestinal integrity in 16 neonatal ECMO patients. METHODS: With 2-day intervals, excretion percentages of lactulose/L-rhamnose (nonmediated diffusion), D-xylose (passive), and 3-O-methyl-D-glucose (active carrier-mediated transport) were measured by gas-liquid chromatography in a 4-hour urine sample. After obtaining baseline data in nine patients, enteral feeding was started in the next seven patients between the third and the ninth day of ECMO. RESULTS: Thirteen patients had increased lactulose/L-rhamnose ratios (>0.05) consistent with increased intestinal permeability. In three patients the lactulose/L-rhamnose ratios were within the normal range. D-xylose excretion percentages were normal (or slightly increased) in 11 patients consistent with normal (or increased) passive carrier-mediated transport. 3-O-methyl-D-glucose excretion percentages were decreased (<10%) in all but one patient, consistent with decreased active carrier-mediated transport. After introduction of enteral nutrition no significant changes of these parameters were seen. CONCLUSIONS: The authors conclude that intestinal integrity is compromised in neonates on ECMO and that introduction of enteral nutrition does not result in further deterioration. This conclusion does not support the practice of withholding enteral nutrition in critically ill newborns supported by ECMO.

Urinary excretion by dogs of intravenously administered simple sugars.

Seven simple sugars, commonly used in intestinal function tests, were simultaneously administered intravenously to dogs and their urinary excretion was measured to assess their potential for metabolic degradation. Lactose, lactulose and 3-O-methyl-D-glucose were excreted unchanged, but small proportions of palatinose and sucrose were not, and were assumed to have been metabolised. More than half of the administered D-xylose and a quarter of L-rhamnose was not excreted and was assumed to have been metabolised. These findings may be significant for the interpretation of differential sugar absorption tests and the D-xylose tolerance test.

Intestinal permeability and absorption in dogs with traumatic injury.

The objectives of this study were to assess the feasibility of using urinary recovery of sugars to evaluate intestinal permeability and absorption in dogs with traumatic injury and to determine if intestinal permeability and absorption are altered in dogs with traumatic injury. After a 6-hour fast, a sugar solution containing lactulose, rhamnose, 3-0-methyl-D-glucose, and xylose was administered via nasoesophageal tube. Urine was collected and quantitated over the 6-hour study period via closed collection urinary catheters. Urinary sugar recoveries were measured by high-pressure anion exchange liquid chromatography and pulsed amperometric detection. Urinary sugar recoveries in the trauma group at 24, 48, and 72 hours after trauma were compared to normal controls. In addition, severity of trauma was compared to urinary sugar recoveries. Twelve client-owned dogs with traumatic injury and 6 healthy controls were enrolled in the study. Lactulose recovery and the lactulose:rhamnose recovery ratio were significantly higher in the trauma group at 48 hours but were no longer different from controls by 72 hours. Xylose recovery was significantly higher in the trauma group when compared to controls at 72 hours, whereas 3-O-methyl-D-glucose recovery was significantly lower in the trauma group at 24 hours. The xylose: 3-O-methyl-D-glucose ratio was higher in the trauma group at all time points. Significant correlation was found between severity of trauma and xylose and 3-O-methyl-D-glucose recoveries 24 hours after injury. Results of this study support the hypothesis that intestinal permeability and absorption are altered in dogs with traumatic injury.

Kinetics and postmucosal effects on urinary recovery of 5 intravenously administered sugars in healthy cats.

The objective of this study was to describe the kinetics of urinary recovery and to evaluate the effects of postmucosal factors on urinary recovery of 5 intravenously administered saccharides. Ten cats received an isotonic sugar solution containing lactulose, rhamnose, xylose, methylglucose, and sucrose intravenously. These sugars were selected because of their prior use for intestinal permeability and mucosal function testing in humans and dogs. Urethral catheterization with a closed collection system was used for collection of cumulative urine samples prior to and 2, 4, 6, 8, 10, 12, and 24 h after administration of the sugar solution. High-pressure anion exchange liquid chromatography with pulsed amperometric detection was used to measure the concentrations of each sugar in the urine and calculate urinary recovery. Twenty-four hour cumulative urinary recovery for each sugar from the cats, was lower than expected compared to dogs and humans. All 5 sugars had the highest percentage of urinary recovery during the first 2 h after administration. Mean sugar elimination rate constants and half-lives ranged from 0.268/h for methylglucose to 0.415/h for lactulose and 1.67 h for lactulose to 2.59 h for methylglucose, respectively. Metabolism and incomplete urine collection are possible reasons for lower cumulative urinary recoveries of these 5 sugars in cats compared with dogs. Although these 5 sugars are not ideal marker molecules, they may still be useful for intestinal permeability and mucosal function testing in cats.

A pilot study examining the relationship among Crohn disease activity, glucagon-like peptide-2 signalling and intestinal function in pediatric patients.

UNLABELLED: BACKGROUND/ OBJECTIVES: The relationship between the enteroendocrine hormone glucagon-like peptide 2 (GLP-2) and intestinal inflammation is unclear. GLP-2 promotes mucosal growth, decreases permeability and reduces inflammation in the intestine; physiological stimulation of GLP-2 release is triggered by nutrient contact. The authors hypothesized that ileal Crohn disease (CD) affects GLP-2 release. METHODS: With ethics board approval, pediatric patients hospitalized with CD were studied; controls were recruited from local schools. Inclusion criteria were endoscopy-confirmed CD (primarily of the small intestine) with a disease activity index >150. Fasting and postprandial GLP-2 levels and quantitative urinary recovery of orally administered 3-O-methyl-glucose (active transport) and lactulose/mannitol (passive) were quantified during the acute and remission phases. RESULTS: Seven patients (mean [± SD] age 15.3 ± 1.3 years) and 10 controls (10.3 ± 1.6 years) were studied. In patients with active disease, fasting levels of GLP-2 remained stable but postprandial levels were reduced. Patients with active disease exhibited reduced glucose absorption and increased lactulose/mannitol recovery; all normalized with disease remission. The change in the lactulose/mannitol ratio was due to both reduced lactulose and increased mannitol absorption. CONCLUSIONS: These findings suggest that pediatric patients with acute ileal CD have decreased postprandial GLP-2 release, reduced glucose absorption and increased intestinal permeability. Healing of CD resulted in normalization of postprandial GLP-2 release and mucosal functioning (nutrient absorption and permeability), the latter due to an increase in mucosal surface area. These findings have implications for the use of GLP-2 and feeding strategies as a therapy in CD patients; further studies of the effects of inflammation and the GLP-2 axis are recommended.

Increased lactulose/rhamnose ratio during fluid load is caused by increased urinary lactulose excretion.

Noninvasive assessment of intestinal permeability in vivo is based on the measurement of urinary excretion of orally administered sugar probes. It is expressed as a ratio, usually lactulose/rhamnose or 3-O-methyl-D-glucose (3-OMG)/rhamnose. In both endotoxemic and control rats that were receiving fluid, we observed an increase in the recovery of lactulose and 3-OMG but not rhamnose in both groups, suggesting an enhancement of intestinal permeability. In the measurement of intestinal permeability, all pre- and postmucosal factors are considered equal for all sugars. We hypothesized that postmucosal factors and not changes in intestinal permeability caused the increased urinary lactulose and 3-OMG recoveries observed during fluid loading. Therefore, the effects of fluid loading on urinary excretion of the sugar probes were studied in healthy rats receiving the sugars intravenously. After intravenous injection, fluid loading increased urinary lactulose recovery threefold but not that of 3-OMG and rhamnose. In conclusion, fluid loading increases the lactulose/rhamnose ratio independent of changes in intestinal permeability. The 3-OMG/rhamnose ratio is not influenced by fluid loading.