The effect of TGF alpha on intestinal solute transport.

The effect of transforming growth factor alpha (TGF alpha) and epidermal growth factor (EGF) on 3-O-methylglucose transport was examined in vitro under short-circuited conditions in stripped rabbit jejunum. Mucosal EGF, 60 ng/ml, stimulated a significant increase in net 3-O-methylglucose transport (Jnet 0.67 +/- 0.15 vs. 0.90 +/- 0.15 microEq/cm2/h; P less than 0.03; n = 6) due to an increased mucosal to serosal flux (Jms 1.2 +/- 0.2 vs. 1.5 +/- 0.2 microEq/cm2/h; P less than 0.03). In contrast, TGF alpha, when applied to both mucosal and serosal surfaces at concentrations of either 60 (n = 6) or 150 (n = 9) ng/ml had no effect on either mucosal to serosal (Jms) or net transport (Jnet) of 3-O-methylglucose. TGF alpha did induce a significant increase in the serosal to mucosal flux (Jsm 60 ng/ml 0.44 +/- 0.02 vs. 0.51 +/- 0.03, 150 ng/ml 0.55 +/- 0.03 vs. 0.64 +/- 0.05 microEq/cm2/h; P less than 0.05). When brush border surface area was examined after exposure to either 60 ng/ml TGF alpha or saline vehicle for 2 h in in vivo isolated jejunal loops no significant difference was found (control 53 +/- 1.9; n = 35 vs. TGF alpha 52 +/- 1.9 microns 2; n = 29). Bioactivity of transforming growth factor alpha was assessed by an gastric acid secretion bioassay and found to be intact. These data provide further evidence for separate and distinct functional roles for these peptides in some biological systems.     

Effects of solvent and solute drag on transmembrane diffusion

The present study compares and quantitates both solvent drag and solute drag forces in a system with both heteropore and homopore membranes. It is shown that tracer solute permeability can be increased if solution flow or driver solute flux is in the direction of tracer diffusion. Either force can decrease tracer permeability if the force can decrease tracer permeability if the force is opposite to the direction of tracer diffusion. The two forces can be additive or one force may reduce the effect of the other force. In the particular system quantitated, solute drag is shown to be some 300 times more effective than solvent drag on a mole-to-mole basis. The use of a number of solute pairs on other homopore and heteropore membranes confirms the finding that the two drag forces can be analyzed or manipulated in a variety of systems.     

Role of intestinal brush border membrane aminopeptidase N in dipeptide transport

The kinetics of uptake of radioactive label from [U-14C]Gly, L-[4,5-3H]Leu and the dipeptide [14C]Gly-L-[4,5-3H]Leu by the brush border membrane vesicles of porcine small intestine have been studied. The effect of aminopeptidase N inhibitors and leucine-binding protein on accumulation rates has also been tested. Comparison of the kinetic parameters for uptake and hydrolysis of Gly-L-Leu makes it possible to conclude that the dipeptide transfer includes two conjugated steps, viz., hydrolysis catalysed by aminopeptidase N and transport of the resultant free amino acids by a specific carrier.     

Role of -SH groups in rat sugar intestinal transport in vivo.

The effect of p-chloromercuribenzoic acid (pCMB), either alone or in the presence of 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), on the 1 mM galactose absorption by in vivo perfused rat intestine has been studied. At 0.25 mM concentration, pCMB inhibits galactose absorption in about 32% but it does not modify the absorption of this sugar when the transport is blocked by 0.5 mM phlorizin, or that of the non-transportable monosaccharide derivative 2-deoxy-D-glucose. This shows that only the active transport component of galactose absorption is inhibited. A 2 min preexposure period is required for the inhibition to appear. The inhibition was not reversed by washing with saline solution even when it contained 0.5 mM dithioerythritol, 10 mM cysteine or 5 and 10 mM EDTA. The simultaneous exposure to 0.25 pCMB and 0.25 mM DTNB inhibits the total galactose entry in about 50%, an effect higher than the one exerted by each reagent separately and close to the one obtained with 0.5 mM phlorizin. Our results, in vivo, confirm the importance of the thiol groups in the cotransport of Na+ and sugar. As DTNB is an SH-reagent of lesser liposolubility than pCMB, the existence of two populations of sulfhydryl groups related to sugar transport which differ in their location within the brushborder membrane and in accessibility from the intestinal lumen, is suggested.     

Role for IL-4 in macromolecular transport across human intestinal epithelium

Increased epithelial permeability is associated with intestinalinflammation, but there is little information on factors that regulatebarrier function in the absence of or before inflammation. We examinedif interleukin (IL)-4, or serum from atopic individuals, could alterthe barrier function of human colonic epithelial (T84) monolayers toantigenic-sized macromolecules. IL-4 and atopic serum significantlydecreased T84 monolayer resistance and increased transepithelialhorseradish peroxidase (HRP) transport. Bidirectional transport studiesdemonstrated that IL-4 selectively enhanced apical-to-basal movement ofHRP. HRP transport induced by IL-4 was inhibited by cold (4°C) andthe tyrosine kinase inhibitor genistein, but not the protein kinase Cinhibitor staurosporine. Electron microscopic analysis demonstratedthat both transcellular and paracellular pathways were affected.Anti-IL-4 antibodies abolished the increase in HRP transport inresponse to both IL-4 and serum. We speculate that enhanced productionof IL-4 in allergic conditions may be a predisposing factor toinflammation by allowing uptake of luminal antigens that gain access tothe mucosal immune system.     

The effect of cyclic deformation and solute binding on solute transport in cartilage

Diffusive transport must play an important role in transporting nutrients into cartilage due to its avascular nature. Recent theoretical studies generally support the idea that cyclic loading enhances large molecule transport through advection. However, to date, reactive transport, i.e. the effects of solute binding, has not yet been taken into consideration in cyclically deformed cartilage. In the present study, we develop a reactive transport model to describe the potential role of binding of solute within cyclically deformed cartilage. Our results show that binding does have a significant effect on transport, particularly for the low IGF-I concentrations typical of synovial fluid. A dynamic loading regime of high strain magnitudes (up to 10%) in combination with high frequencies (e.g. 1 Hz) was seen to produce the most dramatic results with enhanced total uptake ratio as high as 25% averaged over the first 5h of cyclic loading.     

Role of intestinal lymphatics in interstitial volume regulation and transmucosal water transport

Two of the principal functions of intestinal lymphatics are to assist in the maintenance of interstitial volume within relatively normal limits during alterations in capillary filtration (e.g., acute portal hypertension) and the removal of absorbed water and chylomicrons. The contribution of lymphatics to the prevention of interstitial overhydration or dehydration during alterations in transcapillary filtration is similar in the small intestine and colon. While the lymphatics of the small intestine contribute substantially to the removal of absorbed water (particularly at low and moderate absorption rates), the contribution of colonic lymphatics to the removal of the fluid absorbate is negligible. This difference is attributed to the relative caliber and location of lymphatics in the mucosal layer of the small and large intestines. In the small intestine, large lacteals lie in close proximity to transporting epithelium, while colonic lymph vessels are rather sparse and confined to the basal portion of the mucosa. In the small intestine, the lymphatics assume a more important role in removing absorbed water during lipid absorption than during glucose absorption.     

Energy transduction and solute transport in streptococci

Metabolic energy in lactic streptococci can be generated by substrate level phosphorylation and by efflux of end-products in symport with protons. During growth on lactose or glucose Streptococcus cremoris maintains a high proton motive force and phosphate potential. Both energy intermediates dissipate rapidly when the energy supply stops. In the initial phase of starvation the internal phosphoenolpyruvate (PEP) pool increases rapidly and this enables the organism for a prolonged period during starvation to accumulate the energy source via a PEP-dependent uptake system.     

Role of vasa vasorum in transendothelial solute transport in the coronary vessel wall: a study with cryostatic micro-CT

Using cryostatic microscopic computed tomography (micro-CT), we sought to determine the role of coronary vasa vasorum (VV) in transendothelial solute transport in arteries with normal and increased permeability due to high plasma cholesterol levels. In 6-mo-old pigs on a normal (n=23) and 2% high cholesterol (HC) diet (n=8), 2-cm segments of the proximal left anterior descending coronary arteries were removed in vivo after a selective injection of X-ray contrast solution. Harvesting of the specimens occurred at 0, 15, 25, 35, or 45 s after completion of the contrast injection. Specimens were snap frozen and scanned in our cryostatic micro-CT. The spatial distribution of contrast in the coronary artery wall was quantified using the CT images. Right coronary arteries were infused with Microfil to determine VV density (VV/mm2) and the cumulative lumen surface area (mm2/mm3). Transendothelial diffusion of contrast into the coronary vessel wall is a dynamic process starting at both the subintima and the adventitia. The subintimal opacification moves as a wave toward the adventitia, whereas the adventitial wave resolves. The coronary vessel wall in animals on a HC diet shows higher opacification than in normal coronary arteries without an increase of VV total luminal surface area. The loss of endothelial integrity in hypercholesterolemia significantly alters VV solute washin to, and washout from, the coronary artery wall.