Small intestinal epithelial permeability to water-soluble nutrients higher in passerine birds than in rodents.

In the small intestine transcellular and paracellular pathways are implicated in water-soluble nutrient absorption. In small birds the paracellular pathway is quantitatively important while transcellular pathway is much more important in terrestrial mammals. However, there is not a clear understanding of the mechanistic underpinnings of the differences among taxa. This study was aimed to test the hypothesis that paracellular permeability in perfused intestinal segments is higher in passerine birds than rodents. We performed in situ intestinal perfusions on individuals of three species of passerine birds (Passer domesticus, Taeniopygia guttata and Furnarius rufus) and two species of rodents (Mus musculus and Meriones ungiculatus). Using radio-labelled molecules, we measured the uptake of two nutrients absorbed by paracellular and transcellular pathways (L-proline and 3-O-methyl-D-glucose) and one carbohydrate that has no mediated transport (L-arabinose). Birds exhibited ~2 to ~3 times higher L-arabinose clearance per cm2 epithelium than rodents. Moreover, paracellular absorption accounted for proportionally more of 3-O-methyl-D-glucose and L-proline absorption in birds than in rodents. These differences could be explained by differences in intestinal permeability and not by other factors such as increased retention time or higher intestinal nominal surface area. Furthermore, analysis of our results and all other existing data on birds, bats and rodents shows that insectivorous species (one bird, two bats and a rodent) had only 30% of the clearance of L-arabinose of non-insectivorous species. This result may be explained by weaker natural selection for high paracellular permeability in animal- than in plant-consumers. Animal-consumers absorb less sugar and more amino acids, whose smaller molecular size allow them to traverse the paracellular pathway more extensively and faster than glucose.

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

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

The capacity for paracellular absorption in the insectivorous bat Tadarida brasiliensis.

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular processes. The capacity for paracellular absorption seems greater in fliers than in nonfliers, although that conclusion rests mainly on a comparison of flying birds and nonflying mammals because only two frugivorous bat species have been studied. Furthermore, the bats studied so far were relatively large (>85 g, compared with most bat species which are <20 g) and were not insectivores (like about 70 % of bat species). We studied the small (11 g) insectivorous bat Tadarida brasiliensis and tested the prediction that the capacity for paracellular absorption would be as high as in the other bat and avian species studied so far, well above that in terrestrial, nonflying mammals. Using standard pharmacokinetic technique, we measured the extent of absorption (fractional absorption = f) of inert carbohydrate probes: L-arabinose (MM = 150.13) absorbed exclusively by paracellular route and 3OMD-glucose (MM = 194) absorbed both paracellularly and transcellularly. As predicted, the capacity of paracellular absorption in this insectivorous bat was high (L-arabinose f = 1.03 ± 0.14) as in other frugivorous bats and small birds. Absorption of 3OMD-glucose was also complete (f = 1.09 ± 0.17), but >80 % was accounted for by paracellular absorption. We conclude that passive paracellular absorption of molecules of the size of amino acids and glucose is extensive in this bat and, generally in bats, significantly higher than that in nonflying mammals, although the exact extent can be somewhat lower or higher depending on molecule size, polarity and charge.

Identification of a system required for the functional surface localization of sugar binding proteins with class III signal peptides in Sulfolobus solfataricus.

The hyperthermophilic archaeon Sulfolobus solfataricus contains an unusual large number of sugar binding proteins that are synthesized as precursors with a class III signal peptide. Such signal peptides are commonly used to direct archaeal flagellin subunits or bacterial (pseudo)pilins into extracellular macromolecular surface appendages. Likewise, S. solfataricus binding proteins have been suggested to assemble in higher ordered surface structures as well, tentatively termed the bindosome. Here we show that S. solfataricus contains a specific system that is needed for the functional surface localization of sugar binding proteins. This system, encoded by the bas (bindosome assembly system) operon, is composed of five proteins: basABC, three homologues of so-called bacterial (pseudo)pilins; BasE, a cytoplasmic ATPase; and BasF, an integral membrane protein. Deletion of either the three (pseudo)pilin genes or the basEF genes resulted in a severe defect of the cells to grow on substrates which are transported by sugar binding proteins containing class III signal peptides, while growth on glucose and maltose was restored when the corresponding genes were reintroduced in these cells. Concomitantly, DeltabasABC and DeltabasEF cells were severely impaired in glucose uptake even though the sugar binding proteins were normally secreted across the cytoplasmic membrane. These data underline the hypothesis that the bas operon is involved in the functional localization of sugar binding proteins at the cell surface of S. solfataricus. In contrast to surface structure assembly systems of Gram-negative bacteria, the bas operon seems to resemble an ancestral simplified form of these machineries.

Aglycones and sugar moieties alter anthocyanin absorption and metabolism after berry consumption in weanling pigs.

To investigate the absorption and metabolism of anthocyanins (ACNs) with different aglycones and sugar moieties, weanling pigs (11.4 +/- 3.8 kg) were fed, in a single meal, a freeze-dried powder of chokeberry, black currant, or elderberry at a single dose of 229, 140, or 228 mumol total ACN/kg body weight (BW), respectively. These berries provided ACNs with differences in aglycone as well as some unique differences in the sugar moieties. The relative proportions of the different metabolites depended upon concentrations, quantities consumed, and types of glycoside of ACNs in the berry. Delphinidin ACNs were not metabolized to any measurable extent. Cyanidin ACNs were metabolized via methylation and glucuronidation as well as by formation of both derivatives on the same ACN molecule. ACNs with either a di- or trisaccharide attached to them were excreted in the urine primarily as the intact form. Over 80% of the ACN compounds containing rutinose or sambubiose, which were excreted in the urine from black currant, elderberry, or Marion blackberry, were excreted as the intact molecule. The limited metabolism of these ACNs that did occur was via methylation. ACN monoglycosides other than the glucoside were metabolized via methylation and/or glucuronide formation. The monoglucuronide that formed represented a small proportion of the metabolites relative to the methylated or the mixed methylated and glucuronide forms of ACNs. The data clearly demonstrate that the aglycone and the sugar moieties can alter the apparent absorption and metabolism of ACNs.

Digestion and absorption of ferulic acid sugar esters in rat gastrointestinal tract.

We estimated the absorption site and absorptivity of ferulic acid (FA) and its sugar esters, namely 5-O-feruloyl-l-arabinofuranose (FAA) and feruloyl-arabinoxylan (FAXn), in rats on the basis of their recovery in intestinal content and feces by comparing the values with those of a nonabsorbable marker, poly R-478. The results indicated that free FA was absorbed almost completely before reaching cecum. About 40% of dietary FAA was absorbed in rat foregut and 57% disappeared in the cecum. In contrast, about 67% of the FA moiety in FAXn was released and then disappeared predominantly in the hindgut. These results suggested that the existing form of FA in diets affects its absorptivity, its absorption site, and its ensuing fate in the gastrointestinal tract. Those ingested FAs esterified with saccharides; especially, polysaccharides have to transit the hindgut where FA might be released and then absorbed and/or degraded by microflora in lumen. Such microbial degradation may be an important factor affecting the bioavailability of dietary FA.

Intestinal passive absorption of water-soluble compounds by sparrows: effect of molecular size and luminal nutrients.

We tested predictions that: (1) absorption of water-soluble probes decreases with increasing molecular size, consistent with movement through effective pores in epithelia, and (2) absorption of probes is enhanced when measured in the presence of luminal nutrients, as predicted for paracellular solvent drag. Probes (L-arabinose, L-rhamnose, perseitol, lactulose; MW 150.1-342.3 Da) were gavaged in nonanesthetized House sparrows ( Passer domesticus), or injected into the pectoralis, and serially measured in plasma. Bioavailability was calculated as F=AUC by gavage/AUC by injection, where AUC is the area under the curve of plasma probe concentration vs. time. Consistent with predictions, F declined with probe size by 75% from the smallest to the largest probe, and absorption of probes increased by 40% in the presence of luminal glucose or food compared to a mannitol control. Absorption of water-soluble probes by sparrows is much higher than in humans, which is much higher than in rats. These differences seem mainly attributable to differences in paracellular solvent flux and less to differences in effective paracellular pore size.

Passive absorption of hydrophilic carbohydrate probes by the house sparrow Passer domesticus.

To evaluate the permeability of the intestine of the house sparrow Passer domesticus to hydrophilic compounds, we applied a pharmacokinetic technique to measure in vivo absorption of two carbohydrate probes, l-arabinose and d-mannitol. Probes were fed or injected, and blood and excreta were subsequently collected and analyzed by gas chromatography/mass spectrometry. Following injection, plasma probe concentration decreased in a log-linear fashion, implying single-compartment, first-order kinetics. Following oral administration, plasma probe concentrations increased, reached a maximum at 10 min and then decreased in log-linear fashion. Mannitol and arabinose absorption were calculated from the areas under the post-absorption plasma curve and the respective distribution spaces and elimination constants. The amounts absorbed increased linearly with the concentration administered (range 1-1000 mmol x l(-1)), implying a passive process. The mouth-to-cloaca retention time of digesta, measured using the non-absorbable compound potassium ferrocyanide, was independent of probe concentration. On average, 69% of the oral dose of probe was absorbed and this was independent of the concentration of probe administered. This paper supports an earlier report of substantial passive glucose absorption in house sparrows and offers a method to study the extent of hydrophilic solute absorption, which has importance for future research in areas as diverse as biomedical, ecological and evolutionary physiology.

The influence of anesthetic choice, PaCO2, and other factors on osmotic blood-brain barrier disruption in rats with brain tumor xenografts.

UNLABELLED: Increasing the delivery of therapeutic drugs to the brain improves outcome for patients with brain tumors. Osmotic opening of the blood-brain barrier (BBB) can markedly increase drug delivery, but achieving consistent, good quality BBB disruption (BBBD) is essential. We evaluated four experiments compared with our standard isoflurane/O2 protocol to improve the quality and consistency of BBBD and drug delivery to brain tumor and normal brain in a rat model. Success of BBBD was assessed qualitatively with the large molecular weight marker Evans blue albumin and quantitatively by measuring delivery of the low molecular weight marker [3H]-methotrexate. With isoflurane/O2 anesthesia, the effects of two BBBD drugs of different osmolalities were evaluated at two different infusion rates and infusion durations. Arabinose was superior to saline (P = 0.006) in obtaining consistent Evans blue staining in 16 of 24 animals, and it significantly increased [3H]-methotrexate delivery compared with saline in the tumor (0.388 +/- 0.03 vs 0.135 +/-0.04; P = 0.0001), brain around the tumor (0.269 +/- 0.03 vs 0.035 +/- 0.03; P = 0.0001), brain distant to the tumor (0.445 +/- 0.05 vs 0.034 +/- 0.07; P = 0.001), and opposite hemisphere (0.024 +/- 0.00 vs 0.016 +/- 0.00; P = 0.0452). Forty seconds was better than 30 s (P = 0.0372) for drug delivery to the tumor. Under isoflurane/O2 anesthesia (n = 30), maintaining hypocarbia was better than hypercarbia (P = 0.025) for attaining good BBBD. A propofol/ N2O regimen was compared with the isoflurane/O2 regimen, altering blood pressure, heart rate, and PaCO2 as covariates (n = 48). Propofol/N2O was superior to isoflurane/O2 by both qualitative and quantitative measures (P < 0.0001). Neurotoxicity and neuropathology with the propofol/N2O regimen was evaluated, and none was found. These data support the use of propofol/N2O along with maintaining hypocarbia to optimize BBBD in animals with tumors. IMPLICATIONS: Propofol/N2O anesthesia may be better than isoflurane/O2 for optimizing osmotic blood-brain barrier disruption for delivery of chemotherapeutic drugs to brain tumor and normal brain.

Pentose utilization and transport by the ruminal bacterium Prevotella ruminicola.

Plant cell wall polysaccharides are primarily composed of hexose or hexose derivatives, but a significant fraction is hemicellulose which contains pentose sugars. Prevotella ruminicola B14, a predominant ruminal bacterium, simultaneously metabolized pentoses and glucose or maltose, but the organism preferentially fermented pentoses over cellobiose and preferred xylose to sucrose. Xylose and arabinose transport at either low (2 microM) or high (1 mM) substrate concentrations were observed only in the presence of sodium and if oxygen was excluded during the harvest and assay procedures. An artificial electrical potential (delta psi) or chemical gradient of sodium (delta pNa) drove transport in anaerobically prepared membrane vesicles. Because (i) transport was electrogenic, (ii) a delta pNa drove uptake, and (iii) the number of sodium binding sites was approximately 1, it appeared that P. ruminicola possessed pentose/sodium support mechanisms for the transport of arabinose and xylose at low substrate concentrations. Pentose uptake exhibited a low affinity for xylose or arabinose (> 300 microM), and transport of xylose exhibited bi-phasic kinetics which suggested that a second sodium-dependent xylose transport system was present. Little study has been made on solute transport by Prevotella (Bacteroides) species and this work represents the first use of isolated membrane vesicles from these organisms.

Permeability characteristics of deoxygenated sickle cells.

This study investigated the effect of acute deoxygenation on membrane permeability characteristics of sickle cells. Measured fluxes of Na+ and K+ in ouabain-inhibited cells, of chloride and sulfate exchange in 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS)-inhibited and untreated cells, and of erythritol, mannitol, and arabinose in cytochalasin B-inhibited cells indicated that a deoxygenation-induced permeability change occurred in sickle cells only for cations and chloride. Monovalent cation permeabilities increased five-fold, and chloride influx into DIDS treated cells was enhanced nearly threefold on sickle cell deoxygenation. In contrast, no detectable increase in permeability to the other solutes was found. To gain perspective on these findings, similar measurements were performed in normal cells treated with diamide, an agent shown by others to induce a coupled increase in membrane permeability and phospholipid translocation, reminiscent of deoxygenation-induced changes in sickle cells. Although the increase in cation permeability was no greater than that in sickled cells, treatment with 2 mmol/L diamide also produced a twofold increase in the first order rate constants for sulfate exchange and mannitol efflux, indicating a relatively nonselective permeability increase that permitted flux of larger solutes than in the case of deoxygenated sickle cells. These results suggest that the deoxygenation of sickle cells induces a permeability increase that is relatively insensitive to charge, but is restrictive with respect to solute size.

Reversible osmotic opening of the blood-brain barrier in mice.

Unilateral reversible osmotic opening of the blood-brain barrier can be produced in mice. Infusion of 1.8 molal arabinose in water at a rate of 0.64 ml/min for 30 seconds into the internal carotid artery consistently results in ipsilateral brain staining by intravascular Evans blue dye. Osmotic opening is concentration-dependent (threshold, 1.6 molal arabinose) and reversible within 4 hours. No long-term neurologic deficit occurs. These results suggest that reversible osmotic blood-brain barrier opening can be applied to disease models in mice.