A Simple, Robust, and Convenient HPLC Assay for Urinary Lactulose and Mannitol in the Dual Sugar Absorption Test.

BACKGROUND: Heterogeneous laborious analytical methodologies for the determination of urinary lactulose and mannitol limit their utility in intestinal permeability testing. METHODS: We developed an assay using a Shimadzu HPLC system, an Aminex HPX87C column, and refractive index detection. The test was calibrated using a series of dilutions from standard stock solutions of lactulose and mannitol 'spiked' into urine samples. The utility to quantify urinary excretion during the dual sugar absorption test over 6 h was also determined. RESULTS: Lactulose and mannitol were eluted isocratically at 5.7 and 10.1 min, respectively, with water as a mobile phase at a flow rate of 0.3 mL min-1, 858 psi, 60 °C. The calibration curves for both sugars were linear up to 500 µg mL-1 with a limit of detection in standard solutions at 4 µg mL-1 and in 'spiked' urine samples at 15 µg mL-1. The intra-assay and inter-assay CVs were between 2.0-5.1% and 2.0-5.1% for lactulose and 2.5-4.4% and 2.8-3.9% for mannitol. The urinary profiles of the 6 h absorption of lactulose and mannitol showed similar peak-retention times to standard solutions and were well-resolved at 5.9 and 10.4 min, respectively. CONCLUSIONS: The assay was easy to automate, using commonly available equipment and convenient requiring no prior laborious sample derivatization. The simplicity, reproducibility, and robustness of this assay facilitates its use in routine clinical settings for the quantification of intestinal permeability.

Spatiotemporal Mapping of the Contracting Gravid Uterus of the Rabbit Shows Contrary Changes With Increasing Gestation and Dosage With Oxytocin.

Spontaneous and oxytocin induced contractile activity was quantified in the bicornuate uteri of pregnant rabbits maintained in situ, using data from two- and uni- dimensional video spatiotemporal maps (VSTM) of linear and area strain rate and compared statistically. Spontaneous contractions occurred over a range of frequencies between 0.1 and 10 cpm, in gravid animals at 18-21 and at 28 days of gestation, and propagated both radially and longitudinally over the uterine wall overlying each fetus. Patches of contractions were randomly distributed over the entire surface of the cornua and were pleomorphic in shape. No spatial coordination was evident between longitudinal and circular muscle layers nor temporal coordination that could indicate the activity of a localized pacemaker. The density and duration of contractions decreased, and their frequency increased with the length of gestation in the non-laboring uterus. Increasing intravenous doses of oxytocin had no effect on the mean frequencies, or the mean durations of contractions in rabbits of 18-21 days gestation, but caused frequencies to decrease and durations to increase in rabbits of 28 days gestation, from greater spatial and temporal clustering of individual contractions. This was accompanied by an increase in the distance of propagation, the mean size of the patches of contraction, the area of the largest patch of contraction and the overall density of patches. Together these results suggest that progressive smooth muscle hypertrophy and displacement with increasing gestation is accompanied by a decrease in smooth muscle connectivity causing an increase in wall compliance and that oxytocin restores connectivity and decreases compliance, promoting volumetric expulsion rather than direct propulsion of the fetus by peristalsis. The latter effects were reversed by the ß2 adrenergic receptor agonist salbutamol thus reducing area of contraction, and the duration and distance of propagation.

Predicting the viscosity of digesta from the physical characteristics of particle suspensions using existing rheological models.

The measurement of the viscosity of digesta is complicated by settling and compositional changes that accompany digestion. The current work determined whether the apparent and relative viscosities (ηa and ηr) of digesta could be accurately determined from the actual and maximum solid volume fractions (ϕ and ϕmax, respectively) using the Maron-Pierce equation. The rheological properties of digesta from the small intestine of six pigs were determined at a shear rate of 1 s-1 at 37°C. A series of suspensions of plant fibre in a Newtonian liquid (70% aqueous fructose) were made at viscosities similar to pig digesta by adjusting ϕ The relationships between the apparent and relative viscosities (ηa and ηr) and the plant fibre properties; aspect ratio (AR) and ϕ and ϕmax were then determined for digesta and the suspensions. The ARs for the digesta and plant fibre particles were determined using image analysis of scanning electron micrographs and ηa from rheometric flow curves at 37°C, ϕ from image analysis and gas pycnometry, and ϕmax from AR and suspension viscosity. The ηr of pig digesta and the test suspensions calculated using the Maron-Pierce equation were, with the exception of two outliers, in proportion with ηa determined using a rheometer, indicating that ηr could be successfully predicted from the Maron-Pierce equation.

Quantifying Patterns of Smooth Muscle Motility in the Gut and Other Organs With New Techniques of Video Spatiotemporal Mapping.

The uses and limitations of the various techniques of video spatiotemporal mapping based on change in diameter (D-type ST maps), change in longitudinal strain rate (L-type ST maps), change in area strain rate (A-type ST maps), and change in luminous intensity of reflected light (I-maps) are described, along with their use in quantifying motility of the wall of hollow structures of smooth muscle such as the gut. Hence ST-methods for determining the size, speed of propagation and frequency of contraction in the wall of gut compartments of differing geometric configurations are discussed. We also discuss the shortcomings and problems that are inherent in the various methods and the use of techniques to avoid or minimize them. This discussion includes, the inability of D-type ST maps to indicate the site of a contraction that does not reduce the diameter of a gut segment, the manipulation of axis [the line of interest (LOI)] of L-maps to determine the true axis of propagation of a contraction, problems with anterior curvature of gut segments and the use of adjunct image analysis techniques that enhance particular features of the maps.

Spatiotemporal Mapping Techniques Show Clozapine Impairs Neurogenic and Myogenic Patterns of Activity in the Colon of the Rabbit in a Dose-Dependent Manner.

Background: Clozapine, an antipsychotic used in treatment-resistant schizophrenia, has adverse gastrointestinal effects with significant associated morbidity and mortality. However, its effects on defined patterns of colonic contractile activity have not been assessed. Method: We used novel radial and longitudinal spatiotemporal mapping techniques, combined with and monitoring of ambient lumen pressure, in ex vivo preparations of triply and of singly haustrated portions of rabbit colon. We identified the contractile patterns of mass peristalses, fast phasic, and ripple contractions and directly qualified the effects of clozapine, at concentrations of 10 µmol/L, 20 µmol/L, and 30 µmol/L, and of norclozapine, the main metabolite of clozapine, on contractile patterns. The effects of carbachol, serotonin and naloxone on clozapine-exposed preparations were also determined. Tetradotoxin was used to distinguish neurogenic from myogenic contractions. Results: At 10 µmol/L, clozapine temporarily abolished the longitudinal contractile components of mass peristalsis, which on return were significantly reduced in number and amplitude, as was maximal mass peristaltic pressure. These effects were reversed by carbachol (1 µmol/L) and to some extent by serotonin (15 µmol/L). At 10 µmol/L, myogenic ripple contractions were not affected. At 20 µmol/L, clozapine had a similar but more marked effect on mass peristalses with both longitudinal and radial components and corresponding maximal pressure greatly reduced. At 30 µmol/L, clozapine suppressed the radial and longitudinal components of mass peristalses for over 30 min, as well as ripple contractions. Similar dose-related effects were observed on addition of clozapine to the mid colon. At 20 µmol/L, norclozapine had opposite effects to those of clozapine, causing an increase in the frequency of mass peristalsis with slight increases in basal tone. These slightly augmented contractions were abolished on addition of clozapine. Concentrations of norclozapine below 20 µmol/L had no discernible effects. Conclusion: Clozapine, but not norclozapine, has potent effects on the motility of the rabbit colon, inhibiting neurogenic contractions at lower concentrations and myogenic contractions at higher concentrations. This is the likely mechanism for the serious and life-threatening gastrointestinal complications seen in human clozapine-users. These effects appear to be mediated by cholinergic and serotonergic mechanisms. Spatiotemporal mapping is useful in directly assessing the effects of pharmaceuticals on particular patterns of gastrointestinal motility.

The physico-chemical properties of chia seed polysaccharide and its microgel dispersion rheology.

The polysaccharide gel layer surrounding hydrated chia seeds was extracted using water and isolated by ethanol precipitation. The freeze-dried sample consisted of ∼95% non-starch polysaccharides (35% w/w neutral soluble fraction and 65% w/w negatively charged insoluble fraction). The soluble polysaccharide fraction has molar mass, root-mean square radius and intrinsic viscosity of ∼5×10(5)g/mol, 39nm and 719mL/g, respectively. The whole polysaccharide (included soluble and insoluble fractions) when dispersed in water showed presence of irregular shape, fibrous microgel particles with an average size (D4,3) of ∼700µm. Rheological measurements indicated a 'weak' viscoelastic gel and strong shear dependent properties even at low concentration (0.05% w/w). The viscosity of the dispersion was fairly resistant to variations in temperatures (20-80°C), pH (4-12), ionic strengths (0.01-0.5M NaCl) and cation types (MgCl2, CaCl2, NaCl and KCl). The swollen microgel particles dispersed in soluble polysaccharide continuous phase provided complex and potentially useful rheological properties in food systems.

Ex vivo motility in the base of the rabbit caecum and its associated structures: an electrophysiological and spatiotemporal analysis

We examined the coordination between contractile events at different sites in the basal portion of the rabbit caecum and its associated structures that were identified by electrophysiological recordings with simultaneous one-dimensional, and a novel two-dimensional, spatiotemporal mapping technique. The findings of this work provide evidence that the caecum and proximal colon/ampulla coli act reflexly to augment colonic outflow when the caecum is distended and mass peristalsis instituted, the action of the latter overriding the inherent rhythm and direction of haustral propagation in the adjacent portion of the proximal colon but not in the terminal ileum. Further, the findings suggest that the action of the sacculus rotundus may result from its distension with chyme by ileal peristalsis and that the subsequent propagation of contraction along the basal wall of the caecum towards the colon may be augmented by this local distensión (AU)

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Does viscosity or structure govern the rate at which starch granules are digested?

The rates of in vitro digestion of incompletely or fully gelatinised potato and corn starch were measured at 37 °C over 20 min in a rheometer fitted with cup and vane geometry at shear rates of 0.1, 1 and 10 s(-1). Shear rate did not influence the rate of starch digestion provided it was close to physiological levels. However, rates of digestion were significantly reduced when shear rates were below the physiological range (0.1 s(-1)) or when gelatinisation was incomplete. At physiological shear rates the relationship between starch digestion and viscosity was sigmoid in form and following a short initial slow phase a rapid decline in viscosity occurred as starch was digested and the structural integrity of the granules was lost. Conversely, when shear rate was reduced below physiological levels or gelatinisation was incomplete, digestion was hindered, granule integrity was maintained and the relationship between starch and viscosity became linear.

Ex vivo motility in the base of the rabbit caecum and its associated structures: an electrophysiological and spatiotemporal analysis.

We examined the coordination between contractile events at different sites in the basal portion of the rabbit caecum and its associated structures that were identified by electrophysiological recordings with simultaneous one-dimensional, and a novel two-dimensional, spatiotemporal mapping technique. The findings of this work provide evidence that the caecum and proximal colon/ampulla coli act reflexly to augment colonic outflow when the caecum is distended and mass peristalsis instituted, the action of the latter overriding the inherent rhythm and direction of haustral propagation in the adjacent portion of the proximal colon but not in the terminal ileum. Further, the findings suggest that the action of the sacculus rotundus may result from its distension with chyme by ileal peristalsis and that the subsequent propagation of contraction along the basal wall of the caecum towards the colon may be augmented by this local distension.

The Characteristics and Function of Bacterial Lipopolysaccharides and Their Endotoxic Potential in Humans.

Cross-talk between enteral microbiota and human host is essential for the development and maintenance of the human gastrointestinal and systemic immune systems. The presence of lipopolysaccharides (LPS) lysed from the cell membrane of Gram-negative bacteria in the gut lumen is thought to promote the development of a balanced gut immune response whilst the entry of the same LPS into systemic circulation may lead to a deleterious pro-inflammatory systemic immune response. Recent data suggest that chronically low levels of circulating LPS may be associated with the development of metabolic diseases such as insulin resistance, type 2 diabetes, atherosclerosis and cardiovascular disease. This review focuses on the cross-talk between enteral commensal bacteria and the human immune system via LPS. We explain the structural characterisation of the LPS molecule and its function in the bacteria. We then examine how LPS is recognised by various elements of the human immune system and the signalling pathways that are activated by the structure of the LPS molecule and the effect of various concentrations. Further, we discuss the sequelae of this signalling in the gut-associated and systemic immune systems i.e. the neutralisation of LPS and the development of tolerance to LPS.

Assessment of the Effect of Intestinal Permeability Probes (Lactulose And Mannitol) and Other Liquids on Digesta Residence Times in Various Segments of the Gut Determined by Wireless Motility Capsule: A Randomised Controlled Trial.

BACKGROUND: Whilst the use of the mannitol/lactulose test for intestinal permeability has been long established it is not known whether the doses of these sugars modify transit time Similarly it is not known whether substances such as aspirin that are known to increase intestinal permeability to lactulose and mannitol and those such as ascorbic acid which are stated to be beneficial to gastrointestinal health also influence intestinal transit time. METHODS: Gastric and intestinal transit times were determined with a SmartPill following consumption of either a lactulose mannitol solution, a solution containing 600 mg aspirin, a solution containing 500 mg of ascorbic acid or an extract of blackcurrant, and compared by doubly repeated measures ANOVA with those following consumption of the same volume of a control in a cross-over study in six healthy female volunteers. The dominant frequencies of cyclic variations in gastric pressure recorded by the Smartpill were determined by fast Fourier transforms. RESULTS: The gastric transit times of lactulose mannitol solutions, of aspirin solutions and of blackcurrant juice did not differ from those of the control. The gastric transit times of the ascorbic acid solutions were significantly shorter than those of the other solutions. There were no significant differences between the various solutions either in the total small intestinal or colonic transit times. The intraluminal pHs during the initial quartiles of the small intestinal transit times were lower than those in the succeeding quartiles. This pattern did not vary with the solution that was consumed. The power of the frequencies of cyclic variation in intragastric pressure recorded by the Smartpill declined exponentially with increase in frequency and did not peak at the reported physiological frequencies of gastric contractile activity. CONCLUSIONS: Whilst the segmental residence times were broadly similar to those using other methods, the high degree of variation between subjects generally precluded the identification of all but gross variation between treatments. The lack of any differences between treatments in either total small or large intestinal transit times indicates that the solutions administered in the lactulose mannitol test of permeability had no consistent influence on the temporal pattern of absorption. The negatively exponential profile and lack of any peaks in the frequency spectra of cyclic variation in gastric intraluminal pressure that were consistent with reported physiological frequencies of contractile activity profile suggests that the principal source of this variation is stochastic likely resulting from the effects of external events occasioned by normal daily activities on intra-abdominal pressure. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry ACTRN12615000596505.

The effect of fibre and gelatinised starch type on amylolysis and apparent viscosity during in vitro digestion at a physiological shear rate.

An in vitro system was used to determine if the addition of insoluble or soluble fibre to aqueous suspensions of gelatinised starch affected the rate at which the starch was digested. Pre-gelatinised potato or corn starch suspensions were digested with porcine pancreatic amylase in the presence of either finely milled insoluble fibres from various sources or with guar gum. In vitro digestion was conducted at 37°C in a rheometer at a low and constant shear rate of 10s(-1) and the quantity of glucose released measured. The rates of starch digestion and suspension viscosity declined asymptotically and were unaffected by the addition of wheat fibre, but were considerably reduced by the addition of wood and AllBran(®) fibre and to a much greater extent (60%) by the addition of guar. The latter effect may be due to inhibition of amylase activity by non starch polysaccharide sequences.

Characterisation of the contractile dynamics of the resting ex vivo urinary bladder of the pig.

OBJECTIVES: To characterise the area and movements of ongoing spontaneous localised contractions in the resting porcine urinary bladder and relate these to ambient intravesical pressure (Pves ), to further our understanding of their genesis and role in accommodating incoming urine. MATERIALS AND METHODS: We used image analysis to quantify the areas and movements of discrete propagating patches of contraction (PPCs) on the anterior, anterolateral and posterior surfaces of the urinary bladders of six pigs maintained ex vivo with small incremental increases in volume. We then correlated the magnitude of Pves and cyclic changes in Pves with parameters derived from spatiotemporal maps. RESULTS: Contractile movements in the resting bladder consisted only of PPCs that covered around a fifth of the surface of the bladder, commenced at various sites, and were of ≈6 s in duration. They propagated at around 6 mm/s, mainly across the anterior and lateral surface of the bladder by various, sometimes circular, routes in a quasi-stable rhythm, and did not traverse the trigone. The frequencies of these rhythms were low (3.15 cycles/min) and broadly similar to those of cyclic changes in Pves (3.55 cycles/min). Each PPC was associated with a region of stretching (positive strain rate) and these events occurred in a background of more constant strain. The amplitudes of cycles in Pves and the areas undergoing PPCs increased after a sudden increase in Pves but the frequency of cycles of Pves and of origin of PPCs did not change. Peaks in Pves cycles occurred when PPCs were traversing the upper half of the bladder, which was more compliant. The velocity of propagation of PPCs was similar to that of transverse propagation of action potentials in bladder myocytes and significantly greater than that reported in interstitial cells. The size of PPCs, their frequency and their rate of propagation were not affected by intra-arterial dosage with tetrodotoxin or lidocaine. CONCLUSIONS: The origin and duration of PPCs influence both Pves and cyclic variation in Pves . Hence, propagating rather than stationary areas of contraction may contribute to overall tone and to variation in Pves . Spatiotemporal mapping of PPCs may contribute to our understanding of the generation of tone and the basis of clinical entities such as overactive bladder, painful bladder syndrome and detrusor overactivity.

Ascorbic Acid may Exacerbate Aspirin-Induced Increase in Intestinal Permeability.

Ascorbic acid in combination with aspirin has been used to prevent aspirin-induced oxidative GI damage. We aimed to determine whether ascorbic acid reduces or prevents aspirin-induced changes in intestinal permeability over a 6-hr period using saccharidic probes mannitol and lactulose. The effects of administration of 600 mg aspirin alone, 500 mg ascorbic acid alone and simultaneous dosage of both agents were compared in a cross-over study in 28 healthy female volunteers. These effects were also compared with that of a placebo. The ability of ascorbic acid to mitigate the effects of aspirin when administered either half an hour before or after dosage with aspirin was also assessed in 19 healthy female volunteers. The excretion of lactulose over the 6-hr period was augmented after consumption of either aspirin or ascorbic acid compared with that after consumption of placebo. Dosage with ascorbic acid alone augmented the excretion of lactulose more than did aspirin alone. Simultaneous dosage with both agents augmented the excretion of lactulose in an additive manner. The timing of dosage with ascorbic acid in relation to that with aspirin had no significant effect on the excretion of the two sugars. These findings indicate that ascorbic acid does not prevent aspirin-induced increase in gut permeability rather that both agents augment it to a similar extent. The additive effect on simultaneous dosage with both agents in augmenting the absorption of lactulose suggests that each influences paracellular permeability by different pathways.

Aspirin-induced increase in intestinal paracellular permeability does not affect the levels of LPS in venous blood of healthy women.

The presence of subclinical levels of LPS from Gram-negative bacteria, also referred to as endotoxin, in the circulation may induce a pro-inflammatory immune response that leads to the development of obesity and insulin resistance. Recent data indicate that high-fat meals may elevate circulating levels of LPS. However, it is currently unclear how the LPS transits from the gut lumen to the general circulation. We determined whether aspirin-induced damage of the small intestinal mucosa, evidenced by an increase in the paracellular permeability, allows greater transit of LPS into the systemic circulation. The 3-h cumulative excretion of lactulose was significantly increased after the consumption of aspirin solution relative to that after the consumption of an equal volume of water in 15 healthy women (median after aspirin 0.09% of dose vs. median after water 0.03% of dose; P = 0.004). Dosage with aspirin also significantly increased the lactulose : mannitol ratio (median after aspirin 0.014 vs. median after water 0.005; P = 0.017). However, serum LPS levels after the consumption of the aspirin solution were not significantly different from those after consumption of the control (plain water). Further, there was no correlation between body fat content and circulating levels of LPS.

The Limulus Amebocyte Lysate assay may be unsuitable for detecting endotoxin in blood of healthy female subjects.

We examined the factors that may influence the outcome of the Limulus Amebocyte Lysate (LAL) assay, when it is used for quantifying Gram-negative bacterial endotoxin, also referred to as lipopolysaccharide (LPS), in samples of human blood. We found that the method recommended by the manufacturers, based on the reaction time, was inaccurate with any type of serum samples due to the slowing of the initial phase of reaction, likely by serum proteins. We describe an alternative method that is more accurate for use with heated serum samples. Further, we found that components of fresh serum irreversibly sequester endotoxin but that this action may be largely prevented by dilution and heating, but only if this occurs prior to the addition of endotoxin. The tests also indicated that a number of types of proprietary plastic vacutainers appeared to contain significant amounts of endotoxin. However, even when appropriate blood collection containers and calculation methods were used, the levels of endotoxin in serum samples detected by LAL assay were unlikely to reflect the total quantities of endotoxin in that sample and more likely to reflect the capacity of a given serum sample to sequester endotoxin.

Standardising the lactulose mannitol test of gut permeability to minimise error and promote comparability.

BACKGROUND: Lactulose mannitol ratio tests are clinically useful for assessing disorders characterised by changes in gut permeability and for assessing mixing in the intestinal lumen. Variations between currently used test protocols preclude meaningful comparisons between studies. We determined the optimal sampling period and related this to intestinal residence. METHODS: Half-hourly lactulose and mannitol urinary excretions were determined over 6 hours in 40 healthy female volunteers after administration of either 600 mg aspirin or placebo, in randomised order at weekly intervals. Gastric and small intestinal transit times were assessed by the SmartPill in 6 subjects from the same population. Half-hourly percentage recoveries of lactulose and mannitol were grouped on a basis of compartment transit time. The rate of increase or decrease of each sugar within each group was explored by simple linear regression to assess the optimal period of sampling. KEY RESULTS: The between subject standard errors for each half-hourly lactulose and mannitol excretion were lowest, the correlation of the quantity of each sugar excreted with time was optimal and the difference between the two sugars in this temporal relationship maximal during the period from 2½-4 h after ingestion. Half-hourly lactulose excretions were generally increased after dosage with aspirin whilst those of mannitol were unchanged as was the temporal pattern and period of lowest between subject standard error for both sugars. CONCLUSION: The results indicate that between subject variation in the percentage excretion of the two sugars would be minimised and the differences in the temporal patterns of excretion would be maximised if the period of collection of urine used in clinical tests of small intestinal permeability were restricted to 2½-4 h post dosage. This period corresponds to a period when the column of digesta column containing the probes is passing from the small to the large intestine.

Determination of villous rigidity in the distal ileum of the possum (Trichosurus vulpecula).

We investigated the passive mechanical properties of villi in ex vivo preparations of sections of the wall of the distal ileum from the brushtail possum (Trichosurus vulpecula) by using a flow cell to impose physiological and supra-physiological levels of shear stress on the tips of villi. We directly determined the stress applied from the magnitude of the local velocities in the stress inducing flow and additionally mapped the patterns of flow around isolated villi by tracking the trajectories of introduced 3 µm microbeads with bright field micro particle image velocimetry (mPIV). Ileal villi were relatively rigid along their entire length (mean 550 µm), and exhibited no noticeable bending even at flow rates that exceeded calculated normal physiological shear stress (>0.5 mPa). However, movement of villus tips indicated that the whole rigid structure of a villus could pivot about the base, likely from laxity at the point of union of the villous shaft with the underlying mucosa. Flow moved upward toward the tip on the upper portions of isolated villi on the surface facing the flow and downward toward the base on the downstream surface. The fluid in sites at distances greater than 150 µm below the villous tips was virtually stagnant indicating that significant convective mixing in the lower intervillous spaces was unlikely. Together the findings indicate that mixing and absorption is likely to be confined to the tips of villi under conditions where the villi and intestinal wall are immobile and is unlikely to be greatly augmented by passive bending of the shafts of villi.

Characterisation of mixing in the proximal duodenum of the rat during longitudinal contractions and comparison with a fluid mechanical model based on spatiotemporal motility data.

The understanding of mixing and mass transfers of nutrients and drugs in the small intestine is of prime importance in creating formulations that manipulate absorption and digestibility. We characterised mixing using a dye tracer methodology during spontaneous longitudinal contractions, i.e. pendular activity, in 10 cm segments of living proximal duodenum of the rat maintained ex-vivo. The residence time distribution (RTD) of the tracer was equivalent to that generated by a small number (8) of continuous stirred tank reactors in series. Fluid mechanical modelling, that was based on real sequences of longitudinal contractions, predicted that dispersion should occur mainly in the periphery of the lumen. Comparison with the experimental RTD showed that centriluminal dispersion was accurately simulated whilst peripheral dispersion was underestimated. The results therefore highlighted the potential importance of micro-phenomena such as microfolding of the intestinal mucosa in peripheral mixing. We conclude that macro-scale modeling of intestinal flow is useful in simulating centriluminal mixing, whereas multi-scales strategies must be developed to accurately model mixing and mass transfers at the periphery of the lumen.

The partitioning of water in aggregates of undigested and digested dietary particles.

The hydration of fibre particles derived from wheat and wood was quantified, before and after in vitro digestion, and compared with fibre particles from the colonic digesta of pigs and from human faeces. Total water and the extra- and intra-particulate water components were determined using a combination of centrifugation, drying, gas pycnometry and image analysis. The water of saturation (WS) of wood particles and AllBran® measured after in vitro digestion was up to double that of wheat fibres after in vitro digestion, and increased with particle size and loss of soluble material, but was not associated with the chemical composition of the fibres. Fibre that had undergone in vitro gastric digestion and that had been recovered from the colon or faeces, sequestered about 3% of the Ws into intra-particulate spaces, the remainder occupying extra-particulate spaces. The authors speculate that large quantities of fibre must be eaten to sequester toxins that locate into the intra-particulate space.