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PubMed1998

Bowel permeability is improved in Crohn's disease after ileocolectomy.

PURPOSE: Numerous investigators have shown increased bowel permeability in patients with Crohn's disease. It is unclear whether this is a precondition affecting the entire intestine or a consequence of the inflammation and, therefore, only affecting the diseased bowel. The present study tested the hypothesis that resection of diseased bowel in patients with ileocolonic Crohn's disease would correct abnormalities in bowel permeability. METHODS: Ten patients (5 females; mean age, 33 +/- 2 years) with ileocolonic Crohn's disease who underwent elective ileocolic resections had bowel permeability measured preoperatively and postoperatively by the relative urinary clearance of orally consumed lactulose and rhamnose. RESULTS: Mean preoperative bowel permeability in patients with Crohn's disease was significantly elevated relative to healthy volunteers (0.172 +/- 0.04 vs. 0.046 +/- 0.01; P < 0.05, unpaired t-test). After ileocolectomy, bowel permeability decreased in patients with Crohn's disease and reached a normal range by postoperative day 30. CONCLUSIONS: Bowel permeability is increased in patients with ileocolic Crohn's disease because of the presence of diseased bowel and decreases to a normal range in these patients after resection of gross intestinal disease. This suggests that bowel permeability may be a quantitative and clinically effective method with which to assess the presence and severity of diseased bowel in patients with Crohn's disease.

Adult↗
PubMed1995

An in vitro model for endothelial permeability: assessment of monolayer integrity.

An essential component of any in vitro model for endothelial permeability is a confluent cell monolayer. The model reported here utilizes primary human umbilical vein endothelial cells (HUVEC) cultured on recently developed polyethylene terephthalate micropore membranes. Using a modification of the Wright-Giemsa stain, confluent HUVEC monolayers grown on micropore membranes were routinely assessed using light microscopy. Determination of confluence using this method was confirmed by scanning electron microscopy. Transendothelial electrical resistance of HUVEC monolayers averaged 27.9 +/- 11.4 omega.cm2, 10 to 21% higher than literature values. Studies characterizing the permeability of the endothelial cell monolayer to 3H-inulin demonstrated a linear relationship between the luminal concentration of 3H-inulin and its flux across HUVEC monolayers. The slope of the flux versus concentration plot, which represents endothelial clearance of 3H-inulin, was 2.01 +/- 0.076 x 10(-4) ml/min (r2 = .9957). The permeability coefficient for the HUVEC monolayer-micropore membrane barrier was 3.17 +/- 0.427 x 10(-6) cm/s with a calculated permeability coefficient of the HUVEC monolayer alone of 4.07 +/- 0.617 x 10(-6) cm/s. The HUVEC monolayer reduced the permeability of the micropore membrane alone to 3H-inulin (1.43 +/- 0.445 x 10(-5) cm/s) by 78%. Evans blue dye-labeled bovine serum albumin could not be detected on the abluminal side without disruption of the HUVEC monolayer. These results demonstrate a model for endothelial permeability that can be extensively assessed for monolayer integrity by direct visualization, transendothelial electrical resistance, and the permeability of indicator macromolecules.

Cell Membrane Permeability↗
PubMed1996

pH-induced proton permeability changes of plasma membrane vesicles.

In vivo studies with leaf cells of aquatic plant species such as Elodea nuttallii revealed the proton permeability and conductance of the plasma membrane to be strongly pH dependent. The question was posed if similar pH dependent permeability changes also occur in isolated plasma membrane vesicles. Here we report the use of acridine orange to quantify passive proton fluxes. Right-side out vesicles were exposed to pH jumps. From the decay of the applied DeltapH the proton fluxes and proton permeability coefficients (PH+) were calculated. As in the intact Elodea plasma membrane, the proton permeability of the vesicle membrane is pH sensitive, an effect of internal pH as well as external pH on PH+ was observed. Under near symmetric conditions, i.e., zero electrical potential and zero DeltapH, PH+ increased from 65 x 10(-8) at pH 8.5 to 10(-1) m/sec at pH 11 and the conductance from 13 x 10(-6) to 30 x 10(-4) S/m2. At a constant pHi of 8 and a pHo going from 8.5 to 11, PH+ increased more than tenfold from 2 to 26 x 10(-6) m/sec. The calculated values of PH+ were several orders of magnitude lower than those obtained from studies on intact leaves. Apparently, in plasma membrane purified vesicles the transport system responsible for the observed high proton permeability in vivo is either (partly) inactive or lost during the procedure of vesicle preparation. The residue proton permeability is in agreement with values found for liposome or planar lipid bilayer membranes, suggesting that it reflects an intrinsic permeability of the phospholipid bilayer to protons. Possible implications of these findings for transport studies on similar vesicle systems are discussed.

Acridine Orange↗
PubMed1998

The role of yeast VDAC genes on the permeability of the mitochondrial outer membrane.

In addition to the POR1 gene, which encodes the well-characterized voltage dependent anion-selective channel (YVDAC1) of the mitochondrial outer membrane, the yeast Saccharomyces cerevisiae contains a second gene (POR2) encoding a protein (YVDAC2) with 50% sequence identity to YVDAC1. Mitochondria isolated from yeast cells deleted for the POR1 gene (delta por1) had a profoundly reduced outer membrane permeability as measured by the ability of an intermembrane space dehydrogenase to oxidize exogenously added NADH. Mitochondria missing either YVDAC1 or both YVDAC1 and YVDAC2 showed a 2-fold increase in the rate of NADH oxidation when the outer membrane was deliberately damaged. Mitochondria from parental cells showed only a 10% increase indicating that the outer membrane is highly permeable to NADH. In the absence of YVDAC1, we calculate that the outer membrane permeability to NADH is reduced 20-fold. The low NADH permeability in the presence of YVDAC2 was not due to the low levels of YVDAC2 expression as mitochondria from cells expressing levels of YVDAC2 comparable to those of YVDAC1 in parental cells showed no substantial increase in NADH permeability, indicating a minimal role of YVDAC2 in this permeability. The residual permeability may be due to other pathways because cells missing both genes can still grow on nonfermentable carbon sources. However, YVDAC1 is clearly the major pathway for NADH flux through the outer membrane in these mitochondria.

Genes, Fungal↗
PubMed1999

Intestinal permeability, leaky gut, and intestinal disorders.

A major task of the intestine is to form a defensive barrier to prevent absorption of damaging substances from the external environment. This protective function of the intestinal mucosa is called permeability. Clinicians can use inert, nonmetabolized sugars such as mannitol, rhamnose, or lactulose to measure the permeability barrier or the degree of leakiness of the intestinal mucosa. Ample evidence indicates that permeability is increased in most patients with Crohn's disease and in 10% to 20% of their clinically healthy relatives. The abnormal leakiness of the mucosa in Crohn's patients and their relatives can be greatly amplified by aspirin preadministration. Permeability measurements in Crohn's patients reflect the activity, extent, and distribution of the disease and may allow us to predict the likelihood of recurrence after surgery or medically induced remission. Permeability is also increased in celiac disease and by trauma, burns, and nonsteroidal anti-inflammatory drugs. The major determinant of the rate of intestinal permeability is the opening or closure of the tight junctions between enterocytes in the paracellular space. As we broaden our understanding of the mechanisms and agents that control the degree of leakiness of the tight junctions, we will be increasingly able to use permeability measurements to study the etiology and pathogenesis of various disorders and to design or monitor therapies for their management.

Celiac Disease↗
PubMed1985

In-vitro permeability of porcine oral mucosa after epithelial separation, stripping and hydration.

The permeability of porcine skin, gingiva, floor of mouth and buccal mucosa was measured in perfusion chambers using isotopically-labelled water and horseradish peroxidase. Values obtained for the permeability of the epithelium of each of these regions, after separation from the connective tissue with EDTA, did not differ significantly from those obtained for the intact tissue; however, the connective tissue alone had a permeability 2-8 times greater than that of the whole tissue. Stripping the surface layers of the floor of mouth mucosa increased its permeability to that of connective tissue. These results indicate that the functional permeability barrier of the oral mucosa, like that of skin, is located in the epithelium and occupies the superficial layers. After exposure to an aqueous environment for up to 67 h, the permeability of skin and keratinized oral mucosa showed similar but slight increases whereas that of non-keratinized mucosa showed a more rapid rise. These differences may reflect the different composition of the intercellular permeability barrier in keratinized and non-keratinized oral tissues.

Animals↗
PubMed1976

Ionic permeability of epithelial tissues.

The overall permeability of epithelial tissues to solutes is generally determined by analyzing net or unidirectional transepithelial fluxes in response to transepithelial differences of concentration and/or electrical potential using relations that describe diffusional movements across a single membrane. If the solute is uncharged and diffusional movements are transcellular, the overall transepithelial permeability coefficient is determined by the permeabilities of the two limiting cell membranes combinded in series. However, if the solute is charged and the pathway for transepithelial movement involves diffusional flows across at least two membranes arranged in series (i.e. transcellular transport), the value of the overall transepithelial permeability coefficient determined using relations that describe ionic diffusion across a single membrane is not an accurate measure of the permeabilities of the two limiting membranes combined in series. Further, if ionic diffusion is transcellular, permeability coefficients determined from studies of transepithelial fluxes are not only quantitatively incorrect but can also result in grossly erroneous interpretations of changes in transepithelial permeabilities and faulty inferences regarding the route of transepithelial ionic diffusion.

Cell Membrane Permeability↗
PubMed1980

Blocking by 2,4,6-triaminopyrimidine of increased tight junction permeability induced by acetylcholine in the pancreas.

1. The permeability of the paracellular pathway in the isolated rabbit pancreas has been studied with the aid of 2,4,6-triaminopyrimidine. 2. Addition of 2,4,6-triaminopyrimidine (1--10 mM) to the bathing medium has no effect on the rate of fluid secretion or on protein, Na+, K+, Ca2+ and sucrose concentrations in the secreted fluid. 3. When 1 x 10(-5) M carbachol is also added to the 2,4,6-triaminopyrimidine-containing bathing medium, there is a marked reduction in the increase of the paracellular permeability for sucrose and Ca2+ found upon addition of carbachol alone. The enzyme secretion, induced by carbachol, is not affected. 4. The minimal concentration of 2,4,6-triaminopyrimidine in the bathing medium required to reach its maximal effect on the paracellular permeability is approx. 0.55 mM at pH 7.4. 5. The effect of 2,4,6-triaminopyrimidine on the paracellular permeability after carbachol stimulation is also present when 2,4,6-triaminopyrimidine is added 5 min after the addition of 1 x 10(-5) M carbachol. 6. 2,4,6-Triaminopyrimidine has no effect on the increases in enzyme secretion and sucrose permeability caused by 1 x 10(-8) pancreozymin C octapeptide. 7. 2,4,6-Triaminopyrimidine appears in the secreted fluid at a concentration of 50% of that in the bathing medium. Upon addition of 1 x 10(5) M carbachol this concentration increases up to 80%. 8. These results indicate that: (a) the increased paracellular permeability upon stimulation with carbachol is not caused by the enzyme secretion as such and (b) addition of 2,4,6-triaminopyrimidine prevents the carbachol-induced increase in permeability of a channel in the tight junction complex.

Acetylcholine↗
PubMed1980

A light scattering study on the ion permeabilities of dark-adapted bovine rod outer segment disk membranes.

The ion permeability properties of dark adapted bovine rod outer segment disk membranes were studied using light scattering to monitor osmotic responses of disks to various salts and ionophores. A preparation procedure is presented which provides very fresh rod outer segment material with mostly intact stacked disks, but with perforated plasma membrane. It is shown that in this preparation the disks (or rod sacs) are the only osmotically responding compartments and that these responses can be readily monitored by means of light-scattering techniques. The disk membrane is found under the conditions tested, to possess no measurable permeability to cations Na+, Ca2+, Mg2+ nor the the anions Cl-, Br-, NO3-, SO4(2-), H2PO4- and HPO4(2-). There is a considerable K+ permeability, which can be completely abolished by millimolar amounts of divalent cations. The proton permeability of the disk membrane is found to depend dramatically upon the preparation procedure and duration. The fresher the material used the lower is the proton permeability measured. In our freshest preparations, even after freeze-thawing in liquid nitrogen, the disks exhibit an H+ permeability which is so low that it cannot be measured with the techniques used in this study. Even in mitochondrial or chloroplast membmranes, in which proton gradients and therefore a low proton conductance play an essential role, such low proton permeabilities have not been found. This would suggest that proton gradients across the disk membrane could play an important role in the physiological function of the photoreceptor cell. In summary it can be said that the disk membrane, apparently more than any other natural membrane system studied so far, is capable of retaining ion gradients for extended periods of time.

Animals↗
PubMed1981

Control of passive permeability of Chinese hamster ovary cells by external and intracellular ATP.

External ATP causes passive permeability change in several transformed cells, but not in untransformed cells. We studied the effect of external ATP on the passive permeability of CHO-K1 cells, a transformed clone of Chinese hamster ovary cells. Treatment of the cells with external ATP alone did not produce a permeability change, and this was observed only when a mitochondrial inhibitor, such as rotenone or oligomycin, was present together with ATP. These inhibitors reduced the concentration of intracellular ATP and a permeability change by external ATP was observed when intracellular ATP was decreased more than 70%. This requirement for permeability change of CHO-K1 cells was quite unique, since passive permeability change of other transformed cells so far tested was induced by ATP alone. Treatment of CHO-K1 cells with cyclic AMP analogues increased their sensitivity to external ATP about 2-fold. The roles of external and intracellular ATP in controlling passive permeability are discussed.

Adenosine Triphosphate↗
PubMed1981

A food dye, erythrosine B, increases membrane permeability to calcium and other ions.

A widely used food additive erythrosine B, which has been implicated in minimal brain dysfunction in children was examined for its ability to increase membrane permeability to calcium ions. Planar phospholipid bilayer membranes become permeable to calcium, potassium and chloride ions and when erythrosine B is added to the aqueous phase at concentrations which were used by others to demonstrate effects on neuromuscular preparations. The observed increase in permeability to Ca2+ was of sufficient magnitude that equivalent effects on cells would seriously tax the systems which maintain low cytoplasmic Ca2+ levels. The permeability increase in the lipid bilayer membrane is time dependent and increases with erythrosine B concentration raised to a high power (4 to 7). This indicates that the permeability pathway is generated by the cooperative action of a number of erythrosine molecules. This permeability increases dramatically with increasing transmembrane voltage indicating that cells or organelles bearing potentials across their membranes should be particularly sensitive to the dye. We propose that the neurological effects of erythrosine stem from the increased Ca2+ permeability.

Calcium↗
PubMed1983

Modulation of passive permeability by external ATP and cytoskeleton-attacking agents in cultured mammalian cells.

External ATP causes a passive permeability change in several transformed cells, but not in untransformed cells. We previously demonstrated that in CHO-K1 cells, a transformed clone of Chinese hamster ovary cells, the external ATP-dependent permeability change was induced when the intracellular ATP concentration was reduced by a mitochondrial inhibitor (Kitagawa, T. and Akamatsu, Y. (1981) Biochim. Biophys. Acta 649, 76-82). A permeability change with similar characteristics was also observed when the CHO cells were treated with external ATP and a cytoskeleton-attacking agent such as vinblastine or cytochalasin B. Just like mitochondrial inhibitors, vinblastine could increase the sensitivity of transformed 3T3 cells to external ATP but showed no effect on passive permeability of normal 3T3 cells. However, in contrast with the effect of the mitochondrial inhibitors, the cytoskeleton drugs caused the permeability change with little reduction of intracellular ATP concentration, suggesting different actions of these two kinds of drug on the permeability change. The present results suggest an important role of cytoskeletal structures in controlling the external ATP-dependent permeability change in transformed cells. Possible effects of intracellular ATP on cytoskeletal structures are also discussed.

Adenosine Triphosphate↗
PubMed1985

The mode of action of polyacetylene and thiophene photosensitizers on liposome permeability to glucose.

The mode of action of the two photosensitizers 1-phenylhepta-1,3,5-triyne and alpha-terthienyl on membrane permeability was investigated using liposomes entrapped with glucose as a model membrane system. Upon exposure to UV-A light, alpha-terthienyl, and to a much lesser extent phenylheptatriyne, induced leakage of glucose via a photodynamic mechanism in liposomes which had a high degree of unsaturated fatty acid side chains. Enhanced permeability to glucose in these liposomes due to the action of alpha-terthienyl and phenylheptatriyne involved lipid peroxidation, but neither of the two assays used to monitor lipid peroxidation (malondialdehyde and peroxide formation) was directly correlated with the increase in liposome permeability. In liposomes with highly ordered lipid where the fatty acid side chains are saturated, alpha-terthienyl had no effect on glucose permeability. In contrast, phenylheptatriyne was very effective in increasing glucose permeability in these liposomes via a photodynamic mechanism. Addition of lysophosphatidylcholine, which perturbs the order of lipid packing, to these liposomes, completely inhibited the effect of phenylheptatriyne. Conversely, incorporation of cholesterol which increases lipid order, into egg PC liposomes, enhanced the action of phenylheptatriyne. These data suggest that under UV-A irradiation (a) alpha-terthienyl and phenylheptatriyne enhance permeability in liposomes with a high degree of unsaturation involving lipid peroxidation and (b) phenylheptatriyne enhances membrane permeability through some other mechanism when present in a bilayer with a highly ordered lipid environment.

Alkynes↗
PubMed1979

Modification of permeability of frog perineurium to [14C]-sucrose by stretch and hypertonicity.

An in vitro method has been developed to determine quantitatively the permeability of the perineurium to radiotracers at room temperature. The permeability to [14C]sucrose of the isolated perineurium of the sciatic nerve of the frog, Rana pipiens, was measured at rest length, when the perineurium was stretched and after the perineurium had been subjected to hypertonic treatment. Mean permeability at rest length was calculated to be 5.6 +/- 0.27 (S.E.M., n = 45) X 10(-7) cm/sec, and both stretch and hypertonic treatment increased the permeability. A 10% stretch increased permeability reversibly, whereas a 20% stretch or immersion of the perineurium in a hypertonic bath increased permeability irreversibly. Altered permeability under these conditions might be related to changes in the ultrastruct of tight junctions in the perineurium.

Animals↗
PubMed1991

Effects of injection mechanics, pH of infusate and 6-hydroxydopamine on cerebromicrovascular permeability in rats.

The effects of altering the rate, manner and vehicle used for intracerebral injection upon microvascular permeability were studied in Sprague-Dawley rats employing horseradish peroxidase histochemistry. The volume of vehicle delivered and the site of intracerebral injection were kept constant. In comparison to continuous infusion, vascular permeability was significantly greater following manual (intermittent) injections; however, no differences were found when the infusion rate was decreased 10-fold. Use of a buffered vehicle (Hanks' balanced salt solution) with pH adjusted to 7.4, in contrast to the more commonly used non-buffered vehicle (saline-ascorbate), resulted in significant reductions in permeability. The apparent influence of the agent 6-hydroxydopamine (6-OHDA) on changes in vascular permeability was found to vary depending on the type and pH of the vehicle used for injection. Significantly greater permeability resulted with saline-ascorbate (pH 3.1) as the vehicle when compared to Hanks' balanced salt solution (pH 7.4). Changes in vascular permeability can therefore be produced by varying mechanical and vehicular factors which, in the case of 6-OHDA, far outweigh previously reported permeability changes specifically attributed to this neurotoxin.

Animals↗
PubMed1995

Nitric oxide induces vascular permeability changes in the guinea pig conjunctiva.

The role of nitric oxide (NO) as an inflammatory mediator in the mechanism of increased microvascular permeability was examined in a guinea pig model of allergic conjunctivitis. Topical challenge with antigen, compound 48/80, histamine or platelet activating factor (PAF) resulted in a marked increase of the conjunctival vascular permeability. Vascular permeability was determined by measuring the albumin content in the lavage fluid of the challenged eyes after 30 min. Pretreatment with NG-nitro-L-arginine methyl ester (L-NAME) eyedrops caused a significant inhibition of the clinical score and the vascular permeability after challenge with either antigen, histamine or PAF. Aminoguanidine prophylaxis also resulted in a significant inhibition of both the clinical score and vascular permeability in response to all the used provocative agents except PAF. Our observations indicate that NO is an important factor in the induction of the vascular permeability provoked by histamine but seems to play no role in the mechanism by which PAF exerts increased vascular permeability in the conjunctiva.

Albumins↗
PubMed1993

Is small intestinal permeability really increased in relatives of patients with Crohn's disease?

BACKGROUND: Patients with Crohn's disease have increased intestinal permeability, which may precede the development of clinical disease and be involved in disease pathogenesis. Subsequent studies have suggested that, as a group, first-degree relatives of patients with Crohn's disease do not have significantly increased small intestinal permeability rates. The present study proposes that conventional data analysis, used in these studies, may be inappropriate and has overlooked an important observation. METHODS: Lactulose and mannitol permeabilities were defined in healthy controls and in patients with Crohn's disease and their first-degree relatives. RESULTS: Intestinal permeability in relatives was similar to that in the control group, but a subpopulation had abnormally high permeability rates in the absence of clinical evidence for disease. Raw data from another investigator confirmed this finding in an additional study; consequently, it is concluded that the original hypothesis is still viable. A small proportion of individuals, at high risk of developing Crohn's disease, have increased intestinal permeability. CONCLUSIONS: Increased intestinal permeability may precede clinical manifestations of Crohn's disease.

Crohn Disease↗
PubMed1995

CD45RO expression on circulating CD19+ B cells in Crohn's disease correlates with intestinal permeability.

BACKGROUND/AIMS: Increased intestinal permeability is observed in Crohn's disease and a subset of first-degree relatives. An alteration in isoform expression of the common leukocyte antigen (CD45) is also found in a significant fraction of patients. Because this alteration may be a measure of antigen exposure, the hypothesis of the study was that this alteration would be observed in both patients and relatives of patients with Crohn's disease and that this would correlate with increased intestinal permeability. METHODS: Lactulose and mannitol permeability were defined in healthy controls, patients with Crohn's disease, and their first-degree relatives. Simultaneously, peripheral blood was assayed using flow cytometry for CD45RO expression on CD19+ B cells. RESULTS: A subset of relatives had significantly increased permeability, as did the majority of patients with Crohn's disease. A small fraction of peripheral B cells from controls expressed the CD45 isoform (< 6%). This fraction was significantly increased for patients with Crohn's disease and their relatives. Relatives with no clinical evidence of Crohn's disease were only found to have increased CD45RO expression if they had increased permeability. CONCLUSIONS: Individuals at risk for developing Crohn's disease include a subset with increased intestinal permeability. These people have an associated phenotypic alteration of circulating B cells that is not observed in controls or relatives with normal intestinal permeability.

Adolescent↗