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Preparation of alpha-alumina-supported mesoporous bentonite membranes for reverse osmosis desalination of aqueous solutions.

In this study, mesoporous bentonite clay membranes approximately 2 microm thick were prepared on porous alpha-alumina substrates by a sol-gel method. Nanosized clay particles were obtained from commercial Na-bentonite powders (Wyoming) by a process of sedimentation, washing, and freeze-drying. X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and nitrogen adsorption-desorption were employed for membrane characterization. It was found that the content of solids, concentration of polymer binder, and pH value of the clay colloidal suspension had critical influences on membrane formation during the dip-coating process. The membranes were tested for reverse osmosis separation of a 0.1 M NaCl solution. Both water permeability and Na(+) rejection rate of the supported membranes were comparable to those of the compacted thick membranes reported in the literature. However, due to the drastically reduced membrane thickness, water permeance and flux of the supported membranes were significantly higher than those of the compacted thick membranes. It was also observed that the calcination temperature played a critical role in determining structural stability in water and desalination performance of the clay membrane.

Journal Article↗

The effect of the unstirred layer on human red cell water permeability.

A study has been made of water entry into human red blood cells under an osmotic pressure gradient. The measurements were made using a rapid reaction stop flow apparatus, whose construction, calibration, and performance are described in detail. Red cell volume changes were determined from 90 degrees scattered light. The permeability coefficient for water entry under a relative isosmolar concentration of 1 to 1.5 was found to be 0.22 +/- 0.01 cm(4)/sec osmol, which agrees well with our previously published value. The experiments were also designed to measure the thickness of the unstirred layer around the6 red cells. This was found to be 5.5 +/- 0.8 micro under the present experimental conditions. It is concluded that our previously measured permeability coefficient for water entrance under a diffusion gradient does not require correction on account of the unstirred layer.

Biological Transport↗

Very high water permeability in vasopressin-induced endocytic vesicles from toad urinary bladder.

The regulation of transepithelial water permeability in toad urinary bladder is believed to involve a cycling of endocytic vesicles containing water transporters between an intracellular compartment and the cell luminal membrane. Endocytic vesicles arising from luminal membrane were labeled selectively in the intact toad bladder with the impermeant fluid-phase markers 6-carboxyfluorescein (6CF) or fluorescein-dextran. A microsomal preparation containing labeled endocytic vesicles was prepared by cell scraping, homogenization, and differential centrifugation. Osmotic water permeability was measured by a stopped-flow fluorescence technique in which microsomes containing 50 mM mannitol, 5 mM K phosphate, pH 8.5 were subject to a 60-mM inwardly directed gradient of sucrose; the time course of endosome volume, representing osmotic water transport, was inferred from the time course of fluorescence self-quenching. Endocytic vesicles were prepared from toad bladders with hypoosmotic lumen solution treated with (group A) or without (group B) serosal vasopressin at 23 degrees C, and bladders in which endocytosis was inhibited by treatment with vasopressin at 0-2 degrees C (group C), or with vasopressin plus sodium azide at 23 degrees C (group D). Stopped-flow results in all four groups showed a slow rate of 6CF fluorescence decrease (time constants 1.0-1.7 s for exponential fit) indicating a component of nonendocytic 6CF entrapment into sealed vesicles. However, in vesicles from group A only, there was a very rapid 6CF fluorescence decrease (time constant 9.6 +/- 0.2 ms, SEM, 18 separate preparations) with an osmotic water permeability coefficient (Pf) of greater than 0.1 cm/s (18 degrees C) and activation energy of 3.9 +/- 0.8 kcal/mol (16 kJ/mol). Pf was inhibited reversibly by greater than 60% by 1 mM HgCl2. The rapid fluorescence decrease was absent in vesicles in groups B, C, and D. These results demonstrate the presence of functional water transporters in vasopressin-induced endocytic vesicles from toad bladder, supporting the hypothesis that water channels are cycled to and from the luminal membrane and providing a functional marker for the vasopressin-sensitive water channel. The calculated Pf in the vasopressin-induced endocytic vesicles is the highest Pf reported for any biological or artificial membrane.

Animals↗

The capillary permeability of the rete mirabile of the eel, Anguilla vulgaris L.

Direct measurements of transcapillary exchange during steady-state conditions have been performed in the rete mirabile of the eel. The capillaries were found to be more than 30 times as permeable to THO (PTHO = 33.2 times 10(-5) cm sec-1), ethanol and antipyrine than to K+. The mutual relationship of the permeabilities for K+, Na+, urea and sucrose were similar to that between the corresponding free diffusion coefficients in water. The permeability characteristics did not change when the perfusate contained metabolic inhibitors. Additional of 1 mM/l of albumin lead to a significant transcapillary osmotic flow. NaCl, urea and sucrose, however, did not cause osmotic flow. Based on the quantitative values for permeability and volume flow the capillary pores should be about 1300 A and have a frequency of 1.3 times 10(6) per cm2 of the capillary surface, occupying an area of about 0.1% of the total area. The results obtained in the present investigation raise questions as to the validity of the Pappenheimer single-pore theory for transcapillary transport as well as to the fine structure of the capillary pores. As an alternative to Pappenheimer's theory a multi-pathway model is proposed in order to explain transcapillary transport.

Anguilla↗

Water flow across the walls of single muscle capillaries in the frog, Rana pipiens.

Individual capillaries of the transilluminated frog muscle cutaneous pectoris were perfused with suspensions of human red cells in frog Ringer solution containing 1 g/dl bovine serum albumin. The modified Landis technique (Michel, Mason, Curry & Tooke, 1974) was used to measure hydraulic conductivities of the capillary wall. Sucrose osmotic reflexion coefficients of the capillary wall were measured in four capillaries when the superfusate contained 100 mM-sucrose. All experiments were made at 22-24 degrees C. The hydraulic conductivity of arterial capillaries varied from 0.3 to 1.26 X 10(-7) cm/(s cmH2O) with a mean of 0.79 X 10(-7) cm/(s cmH2O) (six capillaries). The hydraulic conductivities of mid-capillaries varied from 0.43 to 1.86 X 10(-7) cm/(s cmH2O) with a mean value of 0.72 X 10(-7) cm/(s cmH2O) (six capillaries). The mean reflexion coefficient to sucrose was 0.12 +/- 0.05 (S.D.). The measured reflexion coefficients to sucrose conform to the hypothesis that 90% of the transcapillary water flow crosses the capillary wall via the principal hydrophilic pathway. The remaining 10% crosses via an exclusive water pathway. The distribution of water flow is similar to that previously described in frog mesenteric capillaries. The mean value of the hydraulic conductivity of frog muscle capillaries is about one-seventh the mean value of the hydraulic conductivity of frog mesenteric capillaries measured at the same temperature. The result conforms to the hypothesis that only a small fraction (mean 10%) of the area of junctional contact between adjacent endothelial cells is available for water and solute exchange in frog muscle capillaries. The hydraulic and diffusional conductances per unit length of open junction appear to be very similar when muscle capillaries are compared to mesenteric capillaries in the frog. Our results lead us to speculate that structures within the intercellular junctions determine the extent of open junction and may modulate the hydraulic conductivity of both the principal water pathway and the exclusive water pathway.

Animals↗

Convective fluid flow through the paracellular system of Necturus gall-bladder epithelium as revealed by dextran probes.

1. Bidirectional paracellular fluxes using radioactive dextrans as inert molecular probes have been measured across Necturus gall-bladder epithelium during conditions of normal fluid absorption. There is a net flux at all radii analysed (0.4-2.2 nm) in the direction of fluid absorption. 2. The net flux is substantial at all radii within the range. The data extraplate to 2 x 10(-6) cm s-1 at zero probe radius, which is very close to the rate of epithelial fluid absorption. 3. The unstirred layers at the epithelial faces during transport have been determined; their contribution to the net fluxes is negligible. 4. Two possible mechanisms for the net flow of probes are considered: (i) that the probes diffuse across the junctions and are then entrained in a local osmotic flow along the interspaces and subepithelium; (ii) that the probes are entrained in volume flow across the junctions and the emergent solution subsequently passes through the interspaces and subepithelium. Model calculations clearly rule out mechanism (i) in which the maximum net flow obtainable is less than 10% of that observed. In addition the presence of leak paths shunting the junctions is not compatible with the observed fluxes. With mechanism (ii) the net flows are correctly predicted with all the fluid flow being transjunctional. The fluid absorption is therefore entirely paracellular. 5. The slope of the net flow curve shows no apparent change in magnitude over the range of the probe radii, indicating that effectively only one population of convective channels is present with parallel walls separated by about 7.7 nm. This agrees with the width previously determined by electron microscopy. 6. If the fluid absorption is junctional then the cellular route offers little if any relative contribution. The hydraulic conductivity of the junctions is not high enough, or the osmotic permeability of the membranes low enough, to accommodate this by osmosis and therefore the junctional fluid absorption must be non-osmotic.

Absorption↗

Mobility of ions, sugar, and water in the cytoplasm of Xenopus oocytes expressing Na(+)-coupled sugar transporters (SGLT1).

A model was set up to study water transport in membrane proteins expressed in Xenopus oocytes. The model was tested experimentally using human and rabbit Na+-glucose cotransporters (SGLT1), and was used to explain controversies regarding unstirred layer effects. Cotransport of Na+, sugar and water was monitored by two-electrode voltage clamp and online measurements of oocyte volume. The specific resistance of the oocyte cytoplasm was found by means of microelectrodes to be 263 +/- 91 Omega cm (S.D., n = 52), or 2.5 times that of Kulori medium, in agreement with reported values of intracellular ion concentrations and diffusion constants. Osmotically induced volume and resistance changes were compatible with a model of the oocyte in which 37 +/- 17 % (S.D., n = 66) of the intracellular volume acts as a free solution while the remainder is inert, being occupied by organelles, etc. The model explains the results of several types of experiments: rapid changes in rates of water cotransport induced by changes in clamp voltage followed by osmotic equilibration in sugar-free conditions; volume changes induced by Na+ transport via the ionophore gramicidin; and uphill water transport. Ethanol (0.5 %) induced a marked swelling of the oocytes of about 16 pl x s(-1). If the specific inhibitor of SGLT1 phlorizin is added from stock solutions in ethanol, the effect of ethanol obfuscates the effects of the inhibitor. We conclude that the transport parameters derived for water cotransport by the SGLT1 can be attributed to the protein residing in the plasma membrane with no significant influences from unstirred layer effects.

Animals↗

Measurement of osmotic reflection coefficient for small molecules in cat hindlimbs.

Capillary osmotic reflection coefficients (sigma) for NaCl, urea, sucrose, and raffinose were measured in the isolated, perfused cat hindlimb using the osmotic transient technique. sigma were determined from the ratio of the maximum rate of transcapillary absorption [delta Jv(max)] to the increase in the osmotic pressure (25-35 mosmol/kg H2O) in the arterial inflow (delta pi a) produced by adding one of the molecules to an albumin-electrolyte perfusate containing isoproterenol (greater than 10(-7) M). delta Jv (max) was determined from organ weight and delta pi a from perfusate osmolalities. For each molecule, the delta Jv(max)/delta pi a ratio increased monotonically with perfusate flow rates (Q) to Q greater than 100 ml.min-1.100 g-1. This ratio was independent of the size of the delta pi a. Apparent sigma values were calculated by dividing these ratios by the capillary hydraulic capacity determined in other studies. At low Q, apparent sigma was comparable to the approximately 0.1 values found by others in skeletal muscle. At the highest Q, apparent sigma for these molecules were at least 0.5. These data are consistent with at least 50% of transcapillary water flow moving through a water-exclusive pathway.

Algorithms↗

Osmotic blood-brain barrier opening to IgM monoclonal antibody in the rat.

Pharmacokinetic parameters of iodinated monoclonal antibody (MAb) delivery to normal rat brain were examined. The mean cerebrovascular permeability-surface area (PA; permeability X capillary surface area) to immunoglobulin M (IgM) MAb (mol wt 1,000,000) 10 min after infusion was 0.40 X 10(-6) S-1. When osmotic blood-brain barrier (BBB) disruption is utilized, the PA increased to 8.36 X 10(-6) S-1. Neither intravenous nor intracarotid MAb administration significantly affected delivery to brain. However, osmotic BBB opening significantly (P less than 0.0005) increased MAb uptake independent of the route of administration. After BBB opening and intracarotid MAb the maximum concentration in brain at 1 h was 0.72% per gram of the total administered dose. For 6 h postdisruption, ipsilateral brain levels were 25- to 100-fold greater than in the contralateral hemisphere or nonbarrier-disrupted controls. MAb concentration in brain slightly decreased over 72 h (P less than 0.05). Antibody recovered from disrupted brain retained 90% of its immunological reactivity for at least 24 h. MAb delivery to ipsilateral brain after BBB disruption was linear over a dose of 0.5-5.0 micrograms IgM, whereas the percentage of the total administered dose remained unchanged. After osmotic treatment, barrier opening was maximal to MAb delivery for 1 min with delivery declining rapidly thereafter. The type of anesthesia used and the administration of a thyroid-blocking agent were found to affect brain MAb levels after BBB disruption.

Animals↗

Nonelectrolyte permeability of canine tracheal epithelium.

We examined the nonelectrolyte permeability characteristics of canine tracheal epithelium in vitro and confirmed that the transepithelial potential difference was 26.3 +/- 2.2 mV, lumen negative. Exposure of the epithelium to a sucrose osmotic load resulted in a streaming potential (SP); a linear relationship was noted between osmotic load and SP. The presence of an osmotic load did not change the short-circuit current and the SP disappeared after removal of the osmotic load. The SP developed with urea, thiourea, D-xylose, and l-glycine were similar to the SP developed for equimolar concentrations of sucrose. The urea and insulin spaces were similar magnitude. When the bulk phase was stirred at 600 rpm, the effective thickness of the unstirred water layer (UWL) external to the epithelial surface was 144 +/- 12 micron. These results support the suggestion that the Staverman reflection coefficients (sigma) of these probe molecules are similar, the estimates of sigma are valid despite the presence of an UWL, and the tracheobronchial epithelium has a pore size smaller than the hydrodynamic radius of urea.

Animals↗