Ammonia absorption from the isolated reticulo-rumen of sheep

In fistulated sheep (50-60 kg live weight) the absorption of ammonia from the reticulo-rumen in vivo was studied applying the technique of the temporarily isolated and washed reticulo-rumen. It was found that, at ammonia concentrations between 3 and 18 mM, ammonia efflux and ammonia net absorption were linearly related to the ammonia concentration in the artificial rumen fluid, whereas influx of ammonia nitrogen from endogenous sources remained almost constant. When the concentration of unionized NH3 was changed at the ratio 1:10:76 by varying the pH from 5.8 to 6.8 and 7.7, ammonia net absorption did not reflect the concentration ratio of unionized NH3, indicating either flux of NH4+ ions or titration of NH4+ to NH3 at the absorptive surface. In the experiments with buffer solutions without ammonium salts and extended over 2 h, ammonia concentrations in the artificial rumen fluid increased due to endogenous nitrogen influx and reached levels far beyond the expected plateau concentration of about 2 mM. Labelling of the N pool in the isolated organ by 15N showed that ammonia efflux had almost ceased in these experiments. It is argued that as yet unidentified changes have taken place in the artificial rumen fluid during the experiment, but there is some reason to believe that volatile fatty acid (VFA) absorption was affecting ammonia absorption.     

Na(+) transport across rumen epithelium of hay-fed sheep is acutely stimulated by the peptide IGF-1 in vitro

An energy-rich diet leads to enhanced ruminal Na(+) absorption, which is associated with elevated plasma insulin-like growth factor 1 (IGF-1) levels and an increased number of IGF-1 receptors in rumen papillae. This study examined the in vitro effect of IGF-1 on Na(+) transport across the rumen epithelium of hay-fed sheep, in which the IGF-1 concentration in plasma is lower than in concentrate-fed animals. At concentrations ranging from 20 to 100 μg l(-1), serosal LR3-IGF-1, a recombinant analogue of IGF-1, rapidly (within 30 min) stimulated the mucosal-to-serosal Na(+) flux (J(ms)Na) and consequently the net Na(+) flux (J(net)Na). Compared with controls, J(net)Na increased by about 60% (P < 0.05) following the serosal application of LR3-IGF-1 (20 μg l(-1)). The IGF-1-induced increment of J(ms)Na and J(net)Na was inhibited by mucosal amiloride (1 mmol l(-1)). Neither IGF-1 nor amiloride altered tissue conductance or the short-circuit current of the isolated rumen epithelium. These data support the assumption that the stimulating effect of serosally applied IGF-1 on Na(+) transport across the rumen epithelium is mediated by Na(+)-H(+) exchange (NHE). A further study was performed with cultured rumen epithelial cells and a fluorescent probe (BCECF) to estimate the rate of pH(i) recovery after acid loading. The pH(i) of isolated rumen epithelial cells was 6.43 ± 0.15 after butyrate loading and recovered by 0.26 ± 0.02 pH units (15 min)(-1). Application of LR3-IGF-1 (20 μg l(-1)) significantly increased the rate of pH(i) recovery to 0.33 ± 0.02 pH units (15 min)(-1). Amiloride administration reduced the recovery rate in both control and IGF-1-stimulated cells. These results show, for the first time, that an acute effect of IGF-1 on Na(+) absorption across rumen epithelium results from increased NHE activity. Insulin-like growth factor 1 is thus important for the fast functional adaptation of ruminal Na(+) transport via NHE.     

Influence of osmolality,short chain fatty acids and deoxycholic acid on mucus secretion in the rat colon

Mucus secretion into the rat colon has been measured in situ using a single perfusion technique. Protein, sialic acid and hexose concentrations in the perfusion solution were found to give reliable estimates of mucus output if samples were homogenized prior to analysis. Mucus output as indicated by an increase in the concentration of mucus constituents was higher when the solution was hypotonic (270 mosm·kg^–1) or hypertonic (370 mosm·kg^–1) than when isotonic solutions (320 mosm·kg^–1) were used. The proportion of hexoses and sialic acid to protein was 23 and 14% at low, 23 and 11% at high osmolality, and 21 and 13% when isotonic solutions were used. Deoxycholic acid (DCA, 4 mmol·l^–1) increased the net secretion of mucus constituents 3 fold, whereas short chain fatty acids (SCFA) had no effect. Mucus composition during all treatments did not change significantly, even when stimulated with DCA.When mucus was released from the epithelial surface by previous perfusion with a DCA containing solution, net water and SCFA absorption rates and mucus output were significantly lowered for 2 to 3 h. However, no correlation between mucus secretion and SCFA absorption was found, indicating that a role for mucus as a diffusion barrier to SCFA is unlikely.Mucus output, which indicates the amount of mucus released from the epithelial surface, probably depends on the direction of net water movement, which follows the osmotic gradient between colon lumen and blood.     

Intracellular pH regulation in guinea-pig caecal and colonic enterocytes during and after loading with short-chain fatty acids and ammonia

Absorption of short-chain fatty acids (SCFA) and ammonia implies considerable fluxes of protons across the epithelium of the large intestine. Efficient regulation of intracellular pH (pH(i)) is therefore essential in these cells. The aim of the present study was to examine the effects of SCFA and of ammonia on pH(i), on pH(i) regulation and to characterize the mechanisms involved in pH(i) regulation in surface enterocytes of the guinea-pig caecal and colonic mucosa. Intact epithelia from the caecum and the distal colon were mounted in a microperfusion chamber. pH(i) was measured by fluorescence microscopy using 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). Addition of SCFA or ammonia to the serosal side changed the enterocyte pH(i) markedly, whereby ammonia caused larger changes in pH(i) than SCFA. In contrast, addition of SCFA to the mucosal solution had no effect on pH(i) and ammonia increased pH(i) only slightly. Basolaterally located pH(i) regulation mechanisms, Na(+)-H(+) exchange and Cl(-)-HCO(3)(-) exchange, are involved mainly in returning pH(i) to normal values. It is concluded that, due to apparently lower permeability of the apical membranes, the caecal and colonic epithelium is protected against pH(i) disturbances caused by the naturally high luminal SCFA and NH(3) concentrations. The major regulation mechanisms of pH(i) are located in the basolateral membrane of the enterocytes.     

Ileal HCO3 secretion: relationship to Na and Cl transport and effect of theophylline.

Interrelationships among Na, Cl, and HCO3 transport processes were examined in short-circuited rabbit ileal mucosa. As serosal (HCO3) was increased from 10 to 50 mM (pH from 7.1 to 7.8), net Na absorption decreased from 4.6 to 0.3 mueq/h-cm2, net Cl flux changed from absorption of 0.9 to secretion of 0.9 and a net HCO3 secretion of 3.0 developed. A similar change in net Cl flux was also observed when serosal Pco2 was altered at constant (HCO3). In Cl-free SO4-Ringer, serosal alkalinization produced net HCO3 secretion which was not significantly less than that observed in Cl-containing Ringer. Theophylline caused secretory changes in net Na and Cl fluxes at both 10 and 50 mM serosal (HCO3). Theophylline did not alter net HCO3 flux in Cl-Ringer but increased net HCO3 flux in SO4-Ringer. Total dc conductance was decreased by both serosal alkalinization and theophylline. Shortcircuit current was consistently increased by theophylline but not by serosal alkalinization. The results indicate that ileal ion transport is regulated in part by serosal pH and/or (HCO3) and that resulting changes in Cl and HCO3 transport are coupled one-for-one with changes in Na transport. Furthermore, HCO3 secretion does not require the presence of Cl in the bathing medium.     

Propionate absorption associated with bicarbonate secretion in vitro in the mouse cecum

Short-chain fatty acids (SCFAs) produced by the microbial fermentation of undigested polysaccharide are rapidly absorbed in the large intestine. One proposed mechanism for this SCFA absorption is SCFA/HCO(-)3 exchange. To provide factual evidence for the operation of SCFA/HCO(-)3 exchange, we mounted an isolated mouse cecum in the Ussing chamber and measured the rates of propionate absorption (J(prop(ms))), alkaline secretion (J(OH(sm))) and total CO2 (HCO(-)3+CO2) secretion (J(tCO2(sm))), and the short-circuit current (I(sc)) with the mucosal side bathed with a Cl- and HCO(-)3-free solution. In the presence of propionate only on the mucosal but not in the serosal solution, J(prop(ms)) was larger when the serosal side was bathed with a HCO(-)3/CO2-containing solution than with a HCO(-)3/CO2-free solution. The addition of propionate to the mucosal side caused an increase in J(OH(sm)) and J(tCO2(sm)), the magnitude of these increases both being much greater with the serosal side bathed with the HCO(-)3/CO2-containing solution than with the HCO(-)3/CO2-free solution. Acetazolamide, a carbonic anhydrase inhibitor, largely suppressed HCO(-)3-dependent components of J(prop(ms)), propionate-induced J(OH(sm)), and propionate-induced J(tCO2(sm)). Acetazolamide, however, did not affect I(sc). The HCO(-)3-dependent component of J(prop(ms)) was not inhibited by either lactate or alpha-cyano-4-hydroxycinnamate, a typical substrate and an inhibitor of the monocarboxylate transporter (MCT1), respectively. It is concluded that an electroneutral, carbonic anhydrase-dependent SCFA/HCO(-)3 exchange mechanism was involved in SCFA absorption. The apical membrane protein for this pathway is not MCT1 and remains to be determined.     

Involvement of carbonic anhydrase in ammonia flux across rumen mucosa in vitro.

Carbonic anhydrase activity in mucosa of the sheep rumen wall was completely inhibited by acetazolamide and ethoxyzolamide. This inhibition significantly reduced ammonia flux across mucosal discs in vitro but was ineffective if short-chain fatty acids were present in the bathing solution. The findings are discussed in relation to the mechanism of ruminal ammonia absorption.     

The interaction between cAMP-dependent and cAMP-independent mechanisms in mediating the somatostatin inhibition of insulin secretion in isolated rat pancreatic islets.

To characterize the intracellular mechanisms by which somatostatin modulates the insulin secretion, studies were performed with isolated rat pancreatic islets at 12 mmol l-1 glucose. Somatostatin (0.1-1000 nmol l-1) inhibited the glucose-induced insulin secretion concentration-dependently. Increasing intracellular cAMP concentration either with dibutyryl-cAMP (1 mmol l-1) or by the adenylate cyclase activator forskolin (20 mumol l-1) partly reversed the inhibition by somatostatin (100 nmol l-1). Neither somatostatin (100 nmol l-1) nor dibutyryl-cAMP (1 mmol l-1 were able to affect the low insulin secretion observed in the absence of extracellular Ca2+. To study cAMP-independent mechanisms of somatostatin, the experiments were performed with and without dibutyryl-cAMP (1 mmol l-1) present. Both somatostatin (100 nmol l-1) and the Ca(2+)-channel blocker verapamil (25 mumol l-1) inhibited the insulin secretion both with and without dibutyryl-cAMP present. An additional inhibition of the insulin secretion was observed when somatostatin was combined with verapamil in the absence, but not in the presence of dibutyryl-cAMP. We conclude that somatostatin inhibits the glucose-induced insulin secretion both by cAMP-dependent mechanism which requires extracellular Ca2+, and by cAMP-independent/verapamil-sensitive Ca(2+)-channel-dependent mechanism.     

Gastric fundic inhibition of sugar transport across the intestinal mucosa of guinea-pig

A low molecular weight peptide designated gastric fundic factor (GFF), extracted from porcine fundic mucosa and administered to the serosal surface of mucosal sheets from guinea-pig intestine, decreased the transport of luminal glucose across the sheets by up to 70%. The results show that gastric fundic inhibition of glucose absorption observed in different animal models in vivo can be reproduced in vitro, and suggest that the intestinal mucosa itself is the target for peptide hormone(s) released by the gastric fundic mucosa. Simultaneous transport of -aminoisobutyric acid, a nonmetabolisable amino acid, through the jejunal mucosa was unaffected as was paracellular permeation by inulin. However, amino acid transport was also reduced when GFF was administered to sheets of ileal mucosa. The intestinal mucosal sheet in vitro is a sensitive and convenient model with which to follow the purification of GFF to homogeneity.     

Monocarboxylate transporter 1 (MCT1) plays a direct role in short-chain fatty acids absorption in caprine rumen

Despite the importance of short-chain fatty acids (SCFA) in maintaining the ruminant physiology, the mechanism of SCFA absorption is still not fully studied. The goal of this study was to elucidate the possible involvement of monocarboxylate transporter 1 (MCT1) in the mechanism of SCFA transport in the caprine rumen, and to delineate the precise cellular localization and the level of MCT1 protein along the entire caprine gastrointestinal tract. RT-PCR revealed the presence of mRNA encoding for MCT1 in all regions of the caprine gastrointestinal tract. Quantitative Western blot analysis showed that the level of MCT1 protein was in the order of rumen ≥ reticulum > omasum > caecum > proximal colon > distal colon > abomasum > small intestine. Immunohistochemistry and immunofluorescence confocal analyses revealed widespread immunoreactive positivities for MCT1 in the caprine stomach and large intestine. Amongst the stratified squamous epithelial cells of the forestomach, MCT1 was predominantly expressed on the cell boundaries of the stratum basale and stratum spinosum. Double-immunofluorescence confocal laser-scanning microscopy confirmed the co-localization of MCT1 with its ancillary protein, CD147 in the caprine gastrointestinal tract. In vivo and in vitro functional studies, under the influence of the MCT1 inhibitors, p -chloromercuribenzoate (pCMB) and p -chloromercuribenzoic acid (pCMBA), demonstrated significant inhibitory effect on acetate and propionate transport in the rumen. This study provides evidence, for the first time in ruminants, that MCT1 has a direct role in the transepithelial transport and efflux of the SCFA across the stratum spinosum and stratum basale of the forestomach toward the blood side.     

Effects of N6,2'-O-dibutyryladenosine 3',5'-cyclic monophosphate, adenosine, and of oxotremorine and 3-isobutyl-1-methylxanthine on the electrically evoked [3H]acetylcholine secretion in the guinea-pig ileum myenteric plexus

The guinea-pig ileum longitudinal muscle-myenteric plexus preparation, pre-incubated with [3H]choline, was mounted in an organ bath and superfused with Tyrode's solution. [3H]Acetylcholine secretion was evoked by 150 electrical shocks at 0.5 Hz. N6,2'-O-Dibutyryladenosine 3',5'-cyclic monophosphate (dibutyryl cyclic AMP) enhanced the [3H]acetylcholine secretion in the presence of eserine and the adenosine receptor antagonist 8-phenyltheophylline (10 mumol l-1). Conversely, in the absence of 8-phenyltheophylline the [3H]acetylcholine secretion was reduced by dibutyryl cyclic AMP. In the absence and presence of 8-phenyltheophylline (apparent KD = 12 mumol l-1), adenosine reduced the [3H]acetylcholine secretion to 33% of control (IC50 = 8 mumol l-1) and to 48% of control (IC50 = 14 mumol l-1) respectively. Neither butyrate, dibutyryl cyclic GMP nor guanosine altered the [3H]acetylcholine secretion. Interaction experiments with 3-isobutyl-1-methylxanthine and oxotremorine were done in the absence of eserine, i.e. when oxotremorine is effective. Oxotremorine depressed the fractional secretion of [3H]acetylcholine with a 'maximal inhibition' of 13% of control (IC50 = 10 nmol l-1). In the presence of 3-isobutyl-1-methylxanthine (5 mmol l-1) oxotremorine depressed the secretion to 2% of control with an apparent IC50 value of 0.9 mumol l-1. 3-Isobutyl-I-methylxanthine (0.01-4 mmol l-1) enhanced the fractional secretion of [3H]acetylcholine with a 'maximal enhancement' value of 232% of control (EC50 = 0.19 mmol l-1). The presence of oxotremorine (30 nmol l-1) counteracted, and higher concentrations reversed, the enhancement caused by 3-isobutyl-1-methylxanthine.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of organic acid absorption on bicarbonate transport in rat colon

The absorption of organic anions and the influence of these anions on the movement of HCO ₃ ^– were studied in vivo in rat colon using a perfusion technique. The absorption of short chain fatty acids (SCFA's) such as acetate, propionate, and butyrate was much greater than that of succinate or lactate. With increasing initial concentration of SCFA up to 100 mmol · l^–1, SCFA absorption increased linearly in correspondence with HCO ₃ ^– appearance. FinalpCO₂ level of the perfusion solution with SCFA was the same as the plasma level. Among the SCFA's, no significant differences in absorption or their effects on HCO ₃ ^– appearance were observed. The presence of Na⁺ stimulated SCFA absorption, and the maximum value was obtained at more than 100 mmol · l^–1 of Na⁺.These results suggest that a specific system for HCO ₃ ^– secretion activated by SCFA exists in the colon, and that this system may control the intraluminal pH by the alkalization of intestinal contents.     

Segmental differences of short-chain fatty acid transport across guinea-pig large intestine.

Unidirectional fluxes of short-chain fatty acids (SCFAs) were measured under short-circuit current conditions across guinea-pig caecum, proximal and distal colon. Fluxes increased linearly with concentration. In the caecum with equal concentrations on both sides of the mucosa the serosal-to-mucosal (sm) fluxes were nearly twice the mucosal-to-serosal (ms) fluxes for all SCFAs; in the distal colon ms fluxes were always higher than sm fluxes. Thus, in the caecum the net effect was secretion while in the distal colon net absorption occurred. In caecum, ms fluxes decreased with chain length while sm fluxes were similar for the three fatty acids, acetic, propionic and butyric acid. In the distal colon both unidirectional fluxes increased with chain length. Fluxes across the mucosa of the proximal colon were intermediate to those in caecum and distal colon. A paracellular transport of short-chain fatty acids is not present. The results indicate that other processes are involved in transcellular transport of SCFA besides non-ionic diffusion.     

Water and ion transport by the urinary bladder of the teleost Pseudopleuronectes americanus.

Water and ion transport by the isolated teleost urinary bladder were studied. The transepithelial electrical PD across sac-type bladder preparations was unstable, i.e., initially mucosa positive but becoming more negative with time. Perfused bladders maintained a low mucosa positive PD which was stable. Both Na and Cl appeared to be actively transported from mucosal side (M) to serosal side (S). Voltage clamping the bladder at 0, -50, or +50 mV had almost no effect on active or passive Na or Cl flux in either direction. Na and Cl transport seemed electrically neutral. Fluid absorption (M to S) was directly correlated with absorption of osmotically active solutes. These solutes were almost all Na and Cl. The bladder acidified and secreted K+ into the mucosal fluid. Divalent ions were concentrated in the mucosal fluid as a result of fluid absorption. Although furosemide and ethacrynic acid inhibited ion and water transport by the bladder, ouabain was effective at a much lower concentration. Ouabain (10(-4) M) inhibited active Na transport when applied only to the mucosal or only to the serosal surface. Ouabain abolished the PD only from the serosal surface.     

Influence of ammonia on sodium absorption in rat proximal colon

The influence of ammonia on sodium and chloride fluxes in rat proximal colon was studied in Ussing chamber experiments. Under short-circuit conditions, the proximal colon absorbed sodium and secreted chloride. The presence of ammonia (30 mmol 1(-1) mucosal) diminished Na+ absorption, but had hardly any influence on Cl- fluxes. Blocking the apical Na+/H+ exchanger isoform NHE2 by amiloride or HOE642 diminished absorptive Na+ and Cl- fluxes. In contrast, the NHE3-specific antagonist S3226 was ineffective. Amiloride and HOE642 also inhibited the effect of ammonia on net sodium absorption. In bicarbonate-free buffer solution, ammonia failed to alter the absorptive fluxes of sodium and chloride, but increased the secretory fluxes of Na+ and Cl-. The latter effect was blocked by HOE642. These results suggest that basal NaCl absorption in rat proximal colon depends to a large extent on NHE2. Mucosal ammonium decreases Na+ absorption and this effect can be antagonized by blocking NHE2. This observation suggests that ammonium interacts with the apical Na+/H+ exchanger, thereby diminishing sodium absorption.     

Motivational effect of cholesterol measurement in general practice health checks.

A randomized trial was conducted in five general practices in and around Aylesbury, Buckinghamshire to assess the motivational effect of cholesterol measurement on compliance with advice to reduce dietary fat intake and to stop smoking. The advice was given by practice nurses during health checks for cardiovascular risk factors. A total of 578 patients were recruited to the study and randomized into two groups. Both groups were given the same advice and were followed up after a median of three months, but the intervention group was also given immediate feedback on their cholesterol concentration. Follow up was completed for 88.2% of subjects, and those who were not followed up were assumed not to have changed their behaviour. The mean fall in total cholesterol at follow up was 0.11 mmol l-1 (95% confidence interval 0.03 to 0.18) in the intervention group who were told their cholesterol result and 0.02 mmol l-1 (95% CI -0.06 to 0.10) in the control group who were not. The proportion of smokers who were not smoking at follow up was 10.7% and 10.1% in the two groups, respectively. Patients in the intervention group with an initial total cholesterol level of 6.50 mmol l-1 or greater showed a mean fall of 6.2% in cholesterol level whereas those with an initial cholesterol level of less than 5.20 mmol l-1 experienced a mean increase of 3.6%, but as differences of this magnitude were also seen in the control group they probably reflect regression to the mean rather than an effect of knowledge of cholesterol level.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro primate gastric mucosa: electrical characteristics

Ion transport by the resting, isolated, rhesus gastric mucosa was assessed under conditions of minimal diffusion limitation to oxygen by 1) the substitution of Na+ and Cl- of the bathing solutions with less permeant ions, 2) the drugs amiloride and ouabain, and 3) estimation of net fluxes of 22Na by methods designed to circumvent the problem of poorly matched tissues. The mucosae developed potential differences of 51.3 +/ 3.5 mV, serosal side positive and had conductances of 5.56 +/- 0.30 mS x cm-2. The permeabilities of the tissues to D-mannitol were between 7.80 x 10(-7) and 3.15 x 10(-7) cm x s-1. The relatively high conductance of this epithelium in the absence of significant edge damage and a low (32%) paracellular conductance stems mainly from a passive permeability to Cl-; active absorption of Na+ and active secretion of Cl- contribute equally to the short-circuit current. The mucosal entry step for Na+ is amiloride sensitive, whereas the serosal exit step can be inhibited by ouabain. The entry step for Cl- at the serosal membrane is possibly sodium dependent.     

The transamination of glutamate and aspartate during absorption in vitro by small intestine of chicken, guinea-pig and rat

1. Procedures are described for direct measurement of the extent and rate of transamination of glutamate and aspartate over periods of up to 90 min, during absorption in vitro by the small intestine of chicken, guinea-pig, and rat.2. During absorption of dicarboxylic amino acids by rat small intestinal segments circulated through the lumen in vitro, alanine contributed up to 85% of the amino acids appearing in the fluid secreted at the serosal surface. In guinea-pig and chicken intestine, the proportion of alanine in the secreted amino acids did not exceed 60%.3. For the different species studied, a relationship was found between the extent to which the dicarboxylic amino acids were transaminated to alanine and the total amount of GPT found in other studies to be present in the intestinal mucosa. In both rat and guinea-pig small intestine, the proportion of alanine in the total amino acids appearing at the serosal surface was similar in the jejunum and ileum. The rate of appearance of alanine in serosal fluid was greater in the ileum than in the jejunum of the rat.4. Reasons are given for supposing that for all the species studied there is a limit to the capacity of the small intestinal mucosa to subject free dicarboxylic amino acids to transamination. It is concluded, however, that it is unlikely that this capacity will be exceeded under in vivo conditions.     

Amino acid transport in the goldfish intestine

1. The serosal transfer of the following eight amino acids: threonine, alanine, serine, histidine, valine, methionine, phenylalanine and leucine, was measured using everted sacs of anterior intestine taken from goldfish acclimatized to 8 degrees C and incubated at 25 degrees C.2. All eight amino acids were actively transported and the serosal transfer correlated with the steady potential (P < 0.001) and with the amino acid-evoked potential (P < 0.05) measured on the same preparations.3. The goldfish rectum actively transported alanine and the steady potential was raised when alanine bathed the mucosa of the everted preparation.4. L-aspartic acid was partly transaminated to alanine by the goldfish anterior intestine; the rectum transaminated alanine to an unidentified amino acid which might have been serine, asparagine or glutamine or some mixture of these three.5. It is suggested that L-amino acids increase the ease by which sodium enters the mucosal cell but that it is the rate at which this sodium is transported across the basal membrane which determines the net serosal transfer of amino acids.     

Volumetric change of the lateral ventricles in the human brain following glucose loading

Lateral ventricular volumes were monitored and quantified using accurately registered magnetic resonance images (MRIs) in six healthy individuals 30 min before and up to 4 h after ingestion of a glucose drink. The volume of the lateral ventricles increased by an average (+/- S.E.M.) of 2.4 +/- 0.4% as blood glucose levels rose from 4.8 +/- 0.2 mmol l-1 to 8.4 +/- 0.4 mmol l-1. This was followed by a peak decrease of 5.99 +/- 3.3% below initial fasting volumes as blood glucose levels fell to 5.0 +/- 0.3 mmol l-1. We suggest that the secondary volume decrease demonstrates a homeostatic process of brain volume regulation for which the mechanism remains uncertain.