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Permeability and integrity properties of lecithin-sphingomyelin liposomes.

The properties of multibilayered liposomes formed from mixtures of sphingomyelin and phosphatidylcholine in varying mole ratio (all containing one mole dicetylphosphate per 10 moles of phospholipids) have been studied. The principal findings are: (1) Over the range 0 to 1 mole fraction sphingomyelin the liposomes exhibit multibilayer structure as visualized by electron microscopy using negative staining. (2) The two phospholipids differ in their interaction with dicetylphosphate in a bilayer structure. In mixtures of the two the effect of sphingomyelin is dominant. (3) The ability of sphingomyelin to form osmotically active liposomes depends on its fatty acid's composition. (4) Liposomes of all mole fractions of sphingomyelin are osmotically active if the C24: 1 fatty acid content of sphingomyelin exceeds 10% of the total acyl residues. The degree of osmotic activity, however, depends upon the molar ratio between the two phospholipids. The highest initial rate of water permeability was found for lecithin liposomes. The maximal change of volume by osmotic gradients was obtained for liposomes composed of 1:1 lecithin to sphingomyelin (mole ratio). (5) Permeability to glucose increased with increasing lecithin mole fraction. (6) Liposomes composed of 1:1 lecithin to sphingomyelin have the largest aqueous volume per mole of phospholipid as measured by glucose trapping. (7) The osmotic fragility of liposomes made of sphingomyelin is higher than for those made of lecithin but the highest osmotic fragility was obtained for liposomes containing lecithin and sphingomyelin in 1:1 molar ratio. (8) When the temperature is abruptly lowered to about 2 degrees C, lipsomes formed from phosphatidylcholine release about 20% of trapped glucose during a transient increase in permeability. Liposomes containing 0.5 mole fraction sphingomyelin release about 30% of the trapped glucose under these conditions. Liposomes composed of sphingomyelin alone do not exhibit this phenomenon.

Animals↗

Transepithelial osmolality differences, hydraulic conductivities, and volume absorption in the proximal tubule.

The weight of current evidence indicates that the proximal tubule has a high transepithelial osmotic water permeability in the range of 3500-6000 microns/sec, which is attributable in large part to the high water permeabilities of the cell membranes. Water movement through these membranes may occur through specialized, proteinaceous channels that can be blocked by sulfhydryl reagents. The water channels probably exclude even the smallest solutes and allow only single file movement of the water molecules as do the water channels previously described in the red blood cell and vasopressin-responsive epithelia. If a significant fraction of the water flow also occurs through the junctional complexes, it seems likely that these junctions could be a site for solute solvent coupling which would contribute to solute absorption by solvent drag and which would be responsible for non-unity reflection coefficients for some solutes such as Na+ and Cl-. This possibility is still a matter of vigorous debate. Since the transepithelial water permeability is high, only a very small osmolality difference (1-10 mOsM) is required to drive normally observed rates of volume absorption both in vivo and in vitro. The osmolality difference is produced at least in part by dilution of the luminal fluid and is possibly augmented by the development of interstitial hyperosmolality because of the rapid transport of preferentially absorbed solutes. In the future it is likely that the most important work in this field will relate to the factors that alter transepithelial water permeability and the solute and water permeabilities of the junctional complexes. Investigation in this area is essential in understanding how changes in capillary and interstitial hydrostatic and colloid osmotic pressure may affect volume absorption.

Absorption↗

Kinetics of water transport in sickle cells.

This paper reports the results of stopped-flow studies on differences in the kinetics of osmotic water transport of sickle and normal erythrocytes. The kinetics of inward osmotic water permeability are similar in sickle and normal red blood cells. In contrast, the kinetics of outward water flux are significantly (approx. 38%) decreased in sickle cells. Deoxygenation does not modify the water influx kinetics in either type of cells, but accelerates considerably the rate of water efflux in sickle cells. No significant variation of water transport kinetics was observed in density-separated cell fractions of either type. The results suggest that membrane-associated hemoglobin may decrease the outward water permeability and that in deoxygenated sickle cells the fraction of hemoglobin S near the lipid bilayer is lower than in oxygenated conditions.

Anemia, Sickle Cell↗

Hydrostatic pressures developed by osmotically swelling vesicles bound to planar membranes.

When phospholipid vesicles bound to a planar membrane are osmotically swollen, they develop a hydrostatic pressure (delta P) and fuse with the membrane. We have calculated the steady-state delta P, from the equations of irreversible thermodynamics governing water and solute flows, for two general methods of osmotic swelling. In the first method, vesicles are swollen by adding a solute to the vesicle-containing compartment to make it hyperosmotic. delta P is determined by the vesicle membrane's permeabilities to solute and water. If the vesicle membrane is devoid of open channels, then delta P is zero. When the vesicle membrane contains open channels, then delta P peaks at a channel density unique to the solute permeability properties of both the channel and the membrane. The solute enters the vesicle through the channels but leaks out through the region of vesicle-planar membrane contact. delta P is largest for channels having high permeabilities to the solute and for solutes with low membrane permeabilities in the contact region. The model predicts the following order of solutes producing pressures of decreasing magnitude: KCl greater than urea greater than formamide greater than or equal to ethylene glycol. Differences between osmoticants quantitatively depend on the solute permeability of the channel and the density of channels in the vesicle membrane. The order of effectiveness is the same as that experimentally observed for solutes promoting fusion. Therefore, delta P drives fusion. When channels with small permeabilities are used, coupling between solute and water flows within the channel has a significant effect on delta P. In the second method, an impermeant solute bathing the vesicles is isosmotically replaced by a solute which permeates the channels in the vesicle membrane. delta P resulting from this method is much less sensitive to the permeabilities of the channel and membrane to the solute. delta P approaches the theoretical limit set by the concentration of the impermeant solute.

Hydrostatic Pressure↗

Permeability of hyaluronic acid solutions. The effects of matrix concentration, calcium, and pH.

The permeability of hyaluronic acid solutions was measured in a chamber closed at each end of semi-permeable membranes. Permeability was strongly concentration dependent below 1.0 gm% hyaluronic acid. Small changes in the ionic environment of 1.0 gm% hyaluronic acid solutions also altered permeability. The permeability of such solutions decreased by a factor of 4.7 in the presence of as little as 5 mM calcium ion and showed optimum permeability at pH 7.0. The pH optimum was not noted in hyaluronic acid preparations known to contain only small amounts of protein.

Calcium↗

Studies of the drug permeability and mechanical properties of free films prepared by cellulose acetate pseudolatex coating system.

Free films produced with cellulose acetate (CA) pseudolatex were prepared by the casting method. The effects of plasticizer concentration, drying temperature, and drying time on drug permeability and mechanical properties of free films were investigated by three-factor spherical second-order composite experimental design. The results were analyzed by the multivariable regression method. The experimental results indicated that plasticizer concentration, drying temperature, and drying time had complex effects on free film permeability and mechanical behavior. These results probably arise from the film-forming ability of CA pseudolatex particles at various conditions and the evaporation of plasticizer during the film-forming process.

Algorithms↗

Endothelin-1 inhibits AVP-stimulated osmotic water permeability in rat inner medullary collecting duct.

Endothelin causes diuresis despite an accompanying decrease in glomerular filtration rate and renal plasma flow. Binding sites for endothelin are located not only in glomeruli but also in the inner medulla, possibly in inner medullary collecting ducts (IMCD). To determine whether endothelin has a direct tubular effect, effects of endothelin on water and urea transport were investigated using isolated microperfusion of rat IMCD segments in vitro. Endothelin, at 10(-10) and 10(-8) M, reversibly inhibited 10(-11) M arginine vasopressin (AVP)-stimulated osmotic water permeability (Pf) by 18 and 24%, respectively. Endothelin (10(-8) M) also inhibited Pf by 23% in the presence of a much higher dose of AVP (10(-9) M), whereas endothelin had no effect on Pf in the absence of AVP. On the other hand, 10(-8) M endothelin did not inhibit Pf stimulated by 10(-3) M dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP). Endothelin had no inhibitory effect on AVP-stimulated urea permeability. These data suggest that endothelin can cause diuresis by inhibiting AVP-stimulated Pf in IMCD and that the site of action is previous to cAMP generation.

Animals↗

Gravitational field enhances permeability of biological membranes to sucrose: an experimental refutation of sucrose-space hypothesis.

Isotonic conditions for the integrity of subcellular organelles are shown to be remarkably influenced by the concentration of sucrose present during their isolation by centrifugation. Using the technique of enzyme osmometry, we show that the content of sucrose in synaptosomes reflects nearly total equilibration across the membrane during centrifugation, due to altered permeability of membranes. Presence of sucrose in the matrix space of mitochondria, as demonstrated by enzyme osmometry of matrix enzymes, indicates that the sucrose-space hypothesis is invalid.

Animals↗

Low aquaporin content and low osmotic water permeability of the plasma and vacuolar membranes of a CAM plant Graptopetalum paraguayense: comparison with radish.

Aquaporin facilitates the osmotic water transport across biomembranes and is involved in the transcellular and intracellular water flow in plants. We immunochemically quantified the aquaporin level in leaf plasma membranes (PM) and tonoplast of Graptopetalum paraguayense, a Crassulacean acid metabolism (CAM) plant. The aquaporin content in the Graptopetalum tonoplast was approximately 1% of that of radish. The content was calculated to be about 3 microg mg(-1) of tonoplast protein. The level of PM aquaporin in Graptopetalum was determined to be less than 20% of that of radish, in which an aquaporin was a major protein of the PM. The PM aquaporin was detected in the mesophyll tissue of Graptopetalum leaf by tissue print immunoblotting. The osmotic water permeability of PM and tonoplast vesicles prepared from both plants was determined with a stopped-flow spectrophotometer. The water permeability of PM was lower than that of the tonoplast in both plants. The Graptopetalum PM vesicles hardly showed water permeability, although the tonoplast showed a relatively high permeability. The water permeability changed depending on the assay temperature and was also partially inhibited by a sulfhydryl reagent. Furthermore, measurement of the rate of swelling and shrinking in different mannitol concentrations revealed that the protoplasts of Graptopetalum showed low water permeability. These results suggest that the low content of aquaporins in PM and tonoplast is one of the causes of the low water permeability of GRAPTOPETALUM: The relationship between the water-storage function of succulent leaves of CAM plants and the low aquaporin level is also discussed.

Amino Acid Sequence↗

[Effect of vasopressin on water permeability of epithelial cells in the kidney collecting duct during postnatal ontogenesis in rats].

Desmopressin caused a statistically significant increase in the water permeability of the outer medullary collecting ducts (OMCD) in 22-days old rats. Concentration of specific V2 receptors increased during postnatal period. Comparison of the V2 receptors concentration, mRNA contents, and changes of water permeability in response to desmopressin suggests that parts of transduction mechanism is situated deeper than the receptors, determines the physiological mechanism at the end of weaning period.

Animals↗

Stopped-flow study of anesthetic effect on water-transport kinetics through phospholipid membranes. Interfacial versus lipid core ligands.

We have compared ligand effects between polar and apolar anesthetic molecules upon water transport across phospholipid membranes by kinetic analysis of the osmotic swelling rate, using a stopped-flow technique. Chloroform and 1-hexanol were used as interfacial ligands, and carbon tetrachloride and n-hexane were used as their counterparts, representing lipid core action. Because anesthetics transform the solid-gel membrane into a liquid-crystalline state, and because phospholipid membranes display an anomaly in permeability at the phase transition, dimyristoylphosphatidylcholine vesicles were studied at temperatures above the main phase transition to avoid this anomaly. All these molecules increased the osmotic swelling rate. However, a significant difference was observed in the activation energy, delta Ep, between polar and apolar molecules; delta Ep was almost unaltered by the addition of polar molecules (chloroform and 1-hexanol), whereas it was decreased by apolar molecules (carbon tetrachloride and n-hexane). The obtained results were analyzed in terms of the dissolution-diffusion mechanism for water permeation across the lipid membrane. It is suggested that polar molecules affect water permeability by altering the partition of water between the membrane interior and water phase, and apolar molecules affect it by altering both the partition and the diffusion of water within the membrane interior.

Anesthetics↗

Significance of plasmalemma aquaporins for water-transport in Arabidopsis thaliana.

The plant plasma membrane intrinsic protein, PIP1b, facilitates water transport. These features were characterized in Xenopus oocytes and it has asked whether aquaporins are relevant for water transport in plants. In order to elucidate this uncertainty Arabidopsis thaliana was transformed with an anti-sense construct targeted to the PIP1b gene. Molecular analysis revealed that the anti-sense lines have reduced steady-state levels of PIP1b and the highly homologous PIP1a mRNA. The cell membrane water permeability was analyzed by swelling of protoplasts, which had been transferred into hypotonic conditions. The results indicate that the reduced expression of the specific aquaporins decreases the cellular osmotic water permeability coefficient approximately three times. The morphology and development of the anti-sense lines resembles that of control plants, with the exception of the root system, which is five times as abundant as that of control plants. Xylem pressure measurement suggests that the increase of root mass compensates the reduced cellular water permeability in order to ensure a sufficient water supply to the plant. The results obtained by this study, therefore, clearly demonstrate that aquaporins are important for plant water transport.

Animals↗

Effects of clofibrate on water permeability and short-cicuit current of toad urinary bladder.

The effects of the oral antidiuretic drug clofibrate on water permeability of the urinary bladder of the toad, Bufo marinus, were studied. Clofibrate did not alter control water flow along an osmotic gradient. Water flow increases due to vasopressin were reversibly inhibited by 1 to 10 mM clofibrate from the serosal side. Clofibrate in the serosal bathing medium inhibited water flow increases due to cyclic AMP but not due to theophylline. Serosal clofibrate concentrations less than 1 mM or clofibrate in the mucosal solution (1 mM) was ineffective in inhibiting the osmotic water permeability response to vasopressin. Clofibrate in the serosal bathing medium (1 mM) inhibited base-line short-circuit current but had no effect on vasopressin-stimulated short-circuit current.

Animals↗

The effect of chain length and lipid phase transitions on the selective permeability properties of liposomes.

This paper describes experiments showing the importance of the fatty acid chain length on the barrier properties of liposomal bilayers, prepared from saturated lecithins, under conditions of lateral phase separation. 1. Above the gel to liquid crystalline phase transition temperature, liposomes prepared from saturated lecithins with 14 or more carbon atoms per acyl chain exist as stable bilayers, which are practically impermeable to ions. 2. At temperatures well above the transition temperature dilauroyl phosphatidylcholine liposomes exhibited osmotic shrinkage, which was dependent on the ionic size of the solute used to bring about the osmotic gradient, indicating that the permeation through these less stable bilayers takes place mainly via individual diffusion of the permeating ions. 3. An enhanced release of trapped potassium from liposomes was demonstrated in the vicinity of the transition temperature. The extent of the increase, however, depended strongly on the length of the paraffin chain. 4. From measurements of the shrinkage behaviour of liposomes in the vicinity of the transition temperature it is concluded that the increased permeability decreases with increasing diameter of the permeating ion. This finding implies that the increased permeability at the transition temperature cannot be ascribed to "macroscopic" rupture of the liposomal membrane. The maximum permeability in the vicinity of the Tc is discussed in terms of probability and size distribution of statistical pore formation at the boundaries of liquid and solid domains.

Cations, Monovalent↗

Regulation of water permeability in toad urinary bladder at two barriers.

The effects of prostaglandin synthesis inhibition by naproxen were studied in toad bladder. Luminal membrane water permeability was evaluated both by the frequency of intramembranous particle aggregates in granular cell luminal membrane and by direct assessment of the rate of change of cell volume during perfusion of an anisosmotic solution. Total tissue water permeability was assessed by transbladder osmotic water flow. Inhibition of prostaglandin synthesis caused luminal membrane water permeability to increase much more than expected from tissue permeability measurements. The addition of a very low dose of antidiuretic hormone (ADH) (0.125 mU/ml) during prostaglandin synthesis inhibition increased luminal membrane water permeability to the same level as maximal stimulation with ADH, while tissue water permeability failed to increase proportionately. The results imply the presence of a regulatable barrier to water movement across toad bladder that is distal to the luminal membrane and subject to control by either prostaglandins or ADH.

Animals↗

Osmotic flow caused by chondroitin sulfate proteoglycan across well-defined nuclepore membranes.

Osmotic flows generated by solutions of Swarm rat chondrosarcoma proteoglycan subunit have been analysed using Nuclepore membranes of well-defined straight-through cylindrical pores of known radius rp. Membranes with rp in the range of 27-500 nm were studied. For semipermeable membranes, which are impermeable to the proteoglycan, the flows were consistently related to r2p and not to r4p (Poiseuille's Law) which demonstrates that the flow is a diffusion-controlled process as described previously (R.P.W. Williams and W.D. Comper, J. Phys. Chem. 91 (1987) 3443). We have also identified a characteristic distance, approx. 50% of the average interparticle spacing, out from the pore surface of the membrane over which the proteoglycan has to move to generate flow. The proteoglycan generates similar osmotic permeability coefficients with membranes up to 125 nm in pore diameter which is significantly larger than the average intermolecular distance. These results have been interpreted in terms of the membrane pore recognising dynamic transient aggregates in the proteoglycan solution.

Animals↗

[Testing the osmotic-diffusion properties for the membranous dressing Bioprocess].

In order to determine of the osmotic-diffusive properties of membranous dressing Bioprocess, which is a microfibrous network of homogeneous cellulose produced in biosynthesis by Acetobacter, the hydraulic permeability (Lp), reflection (sigma) and diffusive permeability (omega) coefficients were measured. The values of these coefficients showed that this membrane possess a low selectivity. This amount it easy permeable for solvent (water) as well as for solutions (aqueous solutions of glucose, sucrose, ethanol, NaCl or KCl). Thus, it attend a demands make for polymeric materials used in therapy of scald and ulceration.

Acetobacter↗

Osmotic water permeabilities of cultured, well-differentiated normal and cystic fibrosis airway epithelia.

Current hypotheses describing the function of normal airway surface liquid (ASL) in lung defense are divergent. One theory predicts that normal airways regulate ASL volume by modulating the flow of isosmotic fluid across the epithelium, whereas an alternative theory predicts that ASL is normally hyposmotic. These hypotheses predict different values for the osmotic water permeability (P(f)) of airway epithelia. We measured P(f) of cultures of normal and cystic fibrosis (CF) airway epithelia that, like the native tissue, contain columnar cells facing the lumen and basal cells that face a basement membrane. Xz laser scanning confocal microscopy recorded changes in epithelial height and transepithelial volume flow in response to anisosmotic challenges. With luminal hyperosmotic challenges, transepithelial and apical membrane P(f) are relatively high for both normal and CF airway epithelia, consistent with an isosmotic ASL. Simultaneous measurements of epithelial cell volume and transepithelial water flow revealed that airway columnar epithelial cells behave as osmometers whose volume is controlled by luminal osmolality. Basal cell volume did not change in these experiments. When the serosal side of the epithelium was challenged with hyperosmotic solutions, the basal cells shrank, whereas the lumen-facing columnar cells did not. We conclude that (a) normal and CF airway epithelia have relatively high water permeabilities, consistent with the isosmotic ASL theory, and the capacity to restore water on airway surfaces lost by evaporation, and (b) the columnar cell basolateral membrane and tight junctions limit transepithelial water flow in this tissue.

Bronchi↗