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.

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.

Differential trafficking of saccharidic probes following aspirin in clinical tests of intestinal permeability in young healthy women.

The effects of inflammatory changes on the absorption of different-sized probes and their permeability ratios are poorly understood. The aim of the present study was to determine the effects of a pharmacological agent on the permeability of the gut mucosa to saccharidic probes of larger and smaller molecular weight. Permeability was assessed by half-hourly urinary excretion of a combined dose of d-mannitol, l-rhamnose and lactulose following consumption of a single 600 mg dose of aspirin and compared with a placebo in a cross-over study in 20 healthy female volunteers. The temporal patterns of excretion of all probes were bimodal, being best fitted by polynomial functions. The relatively small early peak was evident for at least 4 h for smaller sugars, but was less evident with lactulose, being overshadowed by a larger second peak. These conclusions were further supported by separate analyses of the segments of the temporal plots between 2.5 and 4 h and between 4.5 and 6 h. The forms of these curves did not change significantly following dosing with aspirin. A greater proportion of the total dose of mannitol than rhamnose was excreted over the collection period. Following the consumption of aspirin, the cumulative rate of excretion of the smaller sugars (i.e. mannitol and rhamnose) was significantly reduced whereas that of lactulose was increased over the 6 h collection period. Aspirin has opposite effects on the absorption of larger and smaller probes, influencing the outcome of the test. These results have important consequences for the design and comparison of clinical tests of permeability.

The bioelectrical basis and validity of gastrointestinal extracellular slow wave recordings.

Gastrointestinal extracellular recordings have been a core technique in motility research for a century. However, the bioelectrical basis of extracellular data has recently been challenged by claims that these techniques preferentially assay movement artifacts, cannot reproduce the underlying slow wave kinetics, and misrepresent the true slow wave frequency. These claims motivated this joint experimental-theoretical study, which aimed to define the sources and validity of extracellular potentials. In vivo extracellular recordings and video capture were performed in the porcine jejunum, before and after intra-arterial nifedipine administration. Gastric extracellular recordings were recorded simultaneously using conventional serosal contact and suction electrodes, and biphasic and monophasic extracellular potentials were simulated in a biophysical model. Contractions were abolished by nifedipine, but extracellular slow waves persisted, with unchanged amplitude, downstroke rate, velocity, and downstroke width (P>0.10 for all), at reduced frequency (24% lower; P=0.03). Simultaneous suction and conventional serosal extracellular recordings were identical in phase (frequency and activation-recovery interval), but varied in morphology (monophasic vs. biphasic; downstroke rate and amplitude: P<0.0001). Simulations demonstrated the field contribution of current flow to extracellular potential and quantified the effects of localised depolarisation due to suction pressure on extracellular potential morphology. In sum, these results demonstrate that gastrointestinal extracellular slow wave recordings cannot be explained by motion artifacts, and are of a bioelectrical origin that is highly consistent with the underlying biophysics of slow wave propagation. Motion suppression is shown to be unnecessary as a routine control in in vivo extracellular studies, supporting the validity of the extant gastrointestinal extracellular literature.

Manipulating digestion with foods designed to change the physical characteristics of digesta.

We explore how foods can be designed to modulate digestion and to promote health by changing the physical properties of digesta. The physical characteristics of digesta are discussed along with their impact on the physiology of digestion with special reference to sites where these characteristics are likely to influence digestive efficiency. Evidence is reviewed regarding the effects of supplementation with viscoactive agents on the flow and mixing of digesta in particular segments of the human gut that, by changing the rheology and liquid permeability of digesta in that segment, influence specific aspects of digestion and absorption.

Quantification of the effects of the volume and viscosity of gastric contents on antral and fundic activity in the rat stomach maintained ex vivo.

AIMS: The aim of this study was to examine the effect of varying the rheological properties of perfusate on the volume and muscular activity of the various compartments of the rat stomach. METHODS: Image analysis was used to quantify the activity of the ex vivo stomach preparations when perfused according to a ramp profile. RESULTS: The area of the fundus increased to a greater extent than that of the body when watery or viscous material was perfused. However, initial distension of the corpus was greater and occurred more rapidly when viscous material was perfused. Only the fundus expanded when perfusion followed the administration of verapamil. The frequency of antrocorporal contractions decreased significantly and the amplitude of antrocorporal contractions increased significantly with increase in gastric volume. The velocity of antrocorporal contractions did not vary with gastric volume but varied regionally in some preparations being faster distally than proximally. Neither the frequency, amplitude or velocity of antrocorporal contractions differed when pseudoplastic rather than watery fluid was perfused. However, the characteristics of antrocorporal contractions changed significantly when the stomach was perfused with material with rheological characteristics that induce different patterns of wall tension to those normally encountered. Hence, the mean frequency and speed of propagation of antrocorporal contractions increased and their direction of propagation became inconstant.

An exploratory analysis of the suitability of diets fed to a flightless insectivore, the North Island brown kiwi (Apteryx mantelli), in New Zealand.

The ingredients and the macro- and micro-nutrient contents of diets that are fed to captive kiwi at seven New Zealand holding facilities were subject to analysis. The nutrient compositions were compared across facilities and with an estimate of the nutrient composition of the diets of wild kiwi based on the intake of various dietary ingredients reported in the literature. A total of 20 ingredients were used, the number and proportion of these ingredients varied greatly between facilities. Six of the diets were based primarily on meat and three comprised a mixture of meat and cat biscuits. Just one included a proprietary insectivore mix. Nutrient content varied greatly between the seven diets, organic matter ranging from 91.8 to 95.6%, crude protein from 41.9 to 62.9%, and crude fat from 9.0 to 28.7% of dry matter content. Large variations were found in the total content and profile of amino acids and of fatty acids of the diets. The nutrient content of all seven diets differed from that of the natural diet of wild kiwi. Hence, the presence of nutrient deficiencies in the current formulations cannot be excluded.

The roles of filtration and expression in the processing of digesta with high solid phase content.

The processes of filtration and expression of suspensions of biological solids are reviewed in the context of vertebrate digestion. We show that the digesta of the brushtail possum and sheep form a contiguous matrix of particles from which a fluid phase can be expressed by the application of pressure. These findings are examined in respect of their possible contribution to phase separation and the sorting of particles within the gastrointestinal tracts of vertebrates. The morphological adaptations and motility of various digestive compartments are related to the processing of digesta with high solid phase content, in particular the formation and dispersion of the particle matrices. A brief description is given of techniques for evaluating the properties of digesta with high solid phase content.

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.

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.

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.

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.

Anthocyanin absorption and antioxidant status in pigs.

The effect of a simultaneous intake of food or flavonoids on anthocyanins absorption and antioxidant status in pigs was investigated. Twelve male pigs at 27.1 +/- 0.7 kg BW fitted with jugular venous cannulae were maintained in individual metabolic crates. The animals were each given one of three dietary treatments in random order: blackcurrant powder (BC) to give a dose of 100 mg total ACNs/kg BW mixed either with water and sugar (Diet A), cereal (Weet-Bix), milk, and sugar (Diet B), or cereal, milk, sugar, and an additional flavonol (rutin, approximately 100 mg/kg BW) (Diet C). The four major anthocyanins of BC, delphinidin-3-glucoside, delphinidin-3-rutinoside, cyanidin-3-glucoside, and cyanidin-3-rutinoside, were identified and quantified by HPLC-PDA in all three diets. In the pig plasma, four peaks with a reversed pattern to those of anthocyanins in the BC extract were detected. The total amount of anthocyanins absorbed was not significantly different between the three diets, but the rate of absorption and subsequent decline was slower following administration of diet B and C than diet A. All three diets increased antioxidant capacity when measured by the FRAP assay but not when measured by the ORAC and non-protein ORAC assay. However, the increase was delayed and did not appear until 4 h after ingestion, at a time when plasma anthocyanin levels had returned to baseline. The present study demonstrates that the simultaneous intake of food or other flavonoids delays the absorption profile for anthocyanins. Our results also suggest that the increase in antioxidant capacity is not due to dietary anthocyanins but may be due to metabolites that result from anthocyanin consumption.

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.

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 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.

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.

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.

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.