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On the large error introduced in the estimate of the density of membrane pores from permeability measurements when diffusion in "unstirred layer" within the cells is disregarded.

After the development of the "black lipid membrane" techniques, studies of the permeability of labeled water and nonelectrolytes across these artificial membranes have yielded permeability constants comparable in magnitude to those obtained from tracer studies of living cell membranes. This general agreement has affirmed the belief that the living cell membranes are indeed closely similar to these bilayer phospholipid membranes. In this report, we draw attention to a hidden assumption behind such comparisons made: the assumption that labeled material passing through the cell membrane barriers instantly reaches diffusion equilibrium inside the cell. The permeability constants to labeled water (and nonelectrolytes) across lipid layers were obtained using setups in which the lipid membrane was sandwiched between aqueous compartments both of which were vigorously stirred. In studies of permeability of living cell membranes only the outside solution was stirred, the intracellular water remained stationary. Yet the calculations of permeability constants of the cell membrane were made with the tacit assumption, that once the labeled materials pass through the cell membrane, they were instantly mixed with the entire cell contents as if a stirrer operating at infinite speed had been present inside the cells. Ignoring this unstirred condition of the intracellular water, in fact, lumped all the real-life delay due to diffusion in the cytoplasm and added it to the resistance to diffusion of the membrane barrier. The result is an estimated membrane permeability to labeled water (and nonelectrolytes) many times slower than it actually is. The present report begins with a detailed analysis of a specific case: tritiated water diffusion from giant barnacle muscle fibers and two non-living models, one real, one imagined.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Membrane↗

Microvascular permeability after cardiopulmonary bypass. An experimental study.

UNLABELLED: Microvascular permeability is presumed to increase during cardiopulmonary bypass, but this has not been directly demonstrated. METHOD: Therefore, a controlled experimental study was performed in dogs in which the permeability ratio of the small intestinal microvasculature was determined. Shortly after 120 minutes of cardiopulmonary bypass (experimental group, n = 7) or a sham procedure (control group, n = 7), the superior mesenteric venous pressure was raised in a stepwise fashion until the intestinal lymph/plasma protein concentration stabilized at a minimum value. DATA: Minimal lymph/plasma concentration ratio (permeability ratio) of the total proteins and each of six protein fractions was greater in dogs that had been on bypass than in control dogs (p = 0.01 for total proteins and less than 0.05 for five of the fractions). The variability in this regard was large in both groups, and in some animals subjected to bypass the permeability ratio was more than twice the maximum value for the control group. The increase in permeability ratio was greater for large molecules. Inferences: Cardiopulmonary bypass results in a variable increase in microvascular permeability. This results primarily from an increase in the size of large pores in the microvascular barrier.

Animals↗

Studies on vascular permeability in hypertension: action of anthocyanosides.

The initial phase of renal hypertension induced by ligature of the abdominal aorta was accompanied by a transient increase in vascular permeability. This permeability increase has not the same intensity in all parts of the organism: it is greater in the skin and in the aorta wall than in the brain vessels. Treatment of rats with a flavonoid-type drug (anthocyanosides of Vaccinium myrtillus) for 12 days before the induction of hypertension kept the blood-brain barrier permeability normal and limited the increase in vascular permeability in the skin and the aorta wall. As previously demonstrated, the collagens of the blood vessel walls play an important role in the control of vascular permeability. Interaction of these collagens with the drug may be partly responsible for the protection against the permeability-increasing action of hypertension observed in the treated animals.

Animals↗

The effect of Ca2+ and acyl coenzyme A:lysophospholipid acyltransferase inhibitors on permeability properties of the liver mitochondrial inner membrane.

We have reported previously that a number of metabolites and toxins which cause Ca2+ release from mitochondria do so by increasing the permeability of the inner membrane. The metabolic basis of this permeability change is proposed to be perturbation of a phospholipid deacylation-reacylation cycle which results in an accumulation of free fatty acids and lysophospholipids (see Broekemeier, K. M., Schmid, P. C., Schmid, H. H. O., and Pfeiffer, D. R. (1985) J. Biol. Chem. 260, 105-113 and references therein). This hypothesis predicts that inhibitors of acyl-CoA:lysophospholipid acyltransferase would be among those agents which increase membrane permeability and that their effects on permeability could occur in the absence of pyridine nucleotide oxidation or of an accumulation of glutathione disulfide. The hypolipidemic drugs WY-14643 and clofibric acid inhibit the mitochondrial acyl-CoA:lysophospholipid acyltransferase and have the predicted effects on mitochondrial permeability properties. The development of increased permeability due to WY-14643 and clofibric acid requires accumulated Ca2+ specifically, is sensitive to inhibitors of phospholipase A2, and results in a pattern of solute release and swelling which is typical of other Ca2+-releasing agents. Neither agent promotes pyridine nucleotide nor sulfhydryl glutathione oxidation in the absence of Ca2+. In addition, the swelling response to hypolipidemic drugs is not significantly inhibited by dithiothreitol. In the presence of Ca2+, both agents promote an accumulation of free fatty acids. The composition of these lipid degradation products suggests that mitochondria treated with hypolipidemic drugs retain an active lysophospholipase whereas this enzyme is inactivated by Ca2+-releasing agents which alter mitochondrial sulfhydryl groups.

Acyltransferases↗

Permeability of the blood-retinal barrier to carboxyfluorescein in eyes with rhegmatogenous retinal detachment.

Outward and inward permeability of carboxyfluorescein across the blood-retinal barrier were measured fluorophotometrically in seven cynomolgus monkey eyes with experimental rhegmatogenous retinal detachment. Probenecid was used to inhibit outward transport of carboxyfluorescein. The outward permeability was 1.98 +/- 0.31 microliter/min in eyes with retinal detachment and 0.84 +/- 0.15 microliter/min in control eyes with vitrectomy alone (P less than 0.01). The inward permeability, determined separately following intravenous injection, was significantly lower than the outward permeability: 0.14 +/- 0.02 microliter/min for eyes with retinal detachment and 0.04 +/- 0.01 microliter/min for control eyes. Since the outward permeability minus the inward permeability in the presence of probenecid represents that fraction of tracer moving due to fluid flow, it may be concluded that outward flow of fluid across the blood-retinal barrier is a substantial contributor to carboxyfluorescein loss from the vitreous cavity following intravitreal injection.

Animals↗

Serum decreases permeability of some compounds from liposomes.

Although serum is generally regarded to increase the permeability of liposomes containing entrapped substances, we found that a low concentration of serum (10%) significantly reduced the permeability of liposomes to the spin label tempocholine chloride and the polar drug methotrexate, although it increased the permeability of the lipid-soluble drug actinomycin D. Liposomes containing sphingomyelin and cholesterol were considerably less permeable than liposomes containing phosphatidylcholine and cholesterol. Although a higher concentration of serum (88%) increased the permeability of liposomes containing either lipid, the amount of tempocholine which had leaked from sphingomyelin-containing liposomes in 88% serum after 50 h at 37 degrees C was only 25%, three times less than that from phosphatidylcholine-containing liposomes. Thus the effect of serum on liposome permeability depends on the compound entrapped as well as the type of lipid used.

Animals↗

Measurement of blood-retinal barrier permeability: a reproducibility study in normal eyes.

Fluorescein penetration into the posterior vitreous depends on plasma-free fluorescein concentration and blood-retinal barrier (BRB) permeability. The reproducibility of two methods of deriving BRB permeability was studied in 19 normal eyes of 14 subjects using vitreous fluorophotometry on two separate occasions. Plasma-free fluorescence was measured at intervals over 1 hr and posterior vitreous fluorescence was measured before (background scan), within 6 min (bolus) and at 60 min (measurement) after intravenous fluorescein (14 mg X kg-1). A computer algorithm subtracted background fluorescence from the measurement scan which was then corrected for signal spread by using a "spread" function derived from the bolus scan. BRB permeability coefficient and vitreous diffusion coefficients were derived by fitting a mathematical model to the plasma and corrected vitreous fluorescence data. A permeability index was also calculated by dividing the area under the vitreous fluorescence by the area under the plasma fluorescence curve. There were no significant differences in the results between right and left eyes. Mean +/- SD values on first and second occasions for all eyes were permeability coefficient: (1.91 +/- 0.94) and (2.08 +/- 0.95) X 10(-7) cm X s-1; diffusion coefficient: (1.33 +/- 0.68) and (1.19 +/- 0.54) X 10(-5) cm2 X s-1; and permeability index: (2.05 +/- 1.03) and (2.11 +/- 1.02) X 10(-7) cm X s-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Na to Cl permeability in newborn rabbit superficial and juxtamedullary proximal convoluted tubules.

It has been previously demonstrated that superficial (SF) versus juxtamedullary (JM) proximal convoluted tubules (PCT) of rabbit have different intrinsic transport characteristics: Na is less permeable than Cl in the latter portions of the SFPCT, while Cl is less permeable than Na throughout the JMPCT. These permeability differences have major influences on the mechanism of salt reabsorption across the proximal tubules. However, both populations of PCT have the same embryological origin. Studies therefore were designed to examine whether two distinct populations of PCT exist at birth or whether a second population of tubules develops with delivery as a result of some unidentified acute change in humoral factor affecting epithelial transport properties. Both morphological and electrophysiological studies were conducted on PCTs from rabbits within 36 h of birth. Both transmission and scanning electron microscopy studies clearly disclosed that SFPCT are less mature than the JMPCT. Also the SFPCT had a lower Na permeability than Cl (0.55 +/- 0.06) while the JMPCT had a higher Na permeability than Cl (1.37 +/- 0.11). Thus these studies demonstrate that intrinsic heterogeneity of PCT is present at birth. Since the SF Na to Cl permeability approximates that of free diffusion these studies suggest that epithelial discrimination in PCT is part of a maturation process.

Animals↗

The Malpighi lecture. Vascular permeability--the consequence of Malpighi's hypothesis.

The permeability of microvessels to water and small hydrophilic solutes varies considerably between different vascular beds and even between individual vessels in the same vascular bed. Provided the tissues are undamaged, the filtering properties of vessel walls to macromolecules are relatively constant and an analysis of the variations in hydraulic conductances and permeabilities to small ions between different endothelia suggests that the channels for hydrophilic molecules are similar in a wide range of vessels. In undamaged tissues, variations in permeability may be accounted for by variations in the frequency of channels per unit area of vessel wall. The permeability of vessel walls to water and small hydrophilic molecules and their ability to sieve macromolecules are consistent with the molecular filter being a lattice of fibrous molecules which covers and fills the channels through the endothelium. Direct and indirect evidence indicate that globular proteins are part of the molecular filter and it is suggested that they bind the fibrous molecules into a more uniform lattice (the fibre matrix) which greatly increases its selectivity. The question of how the contraction of adjacent endothelial cells might widen the intercellular clefts and thus regulate permeability is discussed. Preliminary experiments show that the permeability of frog capillaries can be varied by varying concentrations of Mg2+ (rather than Ca2+) in the presence of the ionophore A23187 and suggest an interesting line of investigation.

Animals↗

Microwaves and the cell membrane. II. Temperature, plasma, and oxygen mediate microwave-induced membrane permeability in the erythrocyte.

Microwaves (2450 MHz) are shown to increase 22Na permeability of rabbit erythrocytes for exposures only within the narrow temperature range of 17.7 to 19.5 degrees C (Tc) which coincides with a nonlinearity in the Arrhenius plot reflecting an apparent membrane phase transition. Significantly, this response is not observed for cholesterol-loaded erythrocyte membranes which exhibit a linear Arrhenius plot and no apparent phase transition at Tc. The permeability increase at Tc is a nonlinear function of absorbed power but is a linear function of the internal electric field strength of the sample and saturates at approximately 400 mW/g and 600 V/m, respectively. The permeability increase was found to be reversible and transient in that immediately following termination of exposure sodium influx is significantly reduced but returns to normal within 60 min. Extracellular factors exert a significant influence on the microwave effect. The presence of plasma markedly potentiates the increase in 22Na permeability at Tc. Oxygen also modulates the microwave effect with relative hypoxia (5 mm Hg) and hyperoxia (760 mm Hg) enhancing the permeability increase. In contrast, the presence of two antioxidants, ascorbic acid or mercaptoethanol, inhibits the effect. These findings raise important questions about the physical and chemical nature of microwave interactions with cell membranes and also shed light on earlier studies reporting either positive or negative effects on membrane permeability.

Animals↗

Permeability of four disposable protective-clothing materials to seven antineoplastic drugs.

The permeability of four types of protective-clothing material to seven injectable antineoplastic drugs was studied. The protective materials tested were Saranex-laminated Tyvek, polyethylene-coated Tyvek, nonporous Tyvek, and Kaycel. Circles 6 cm in diameter were cut from a single garment of each material and exposed to each drug. Permeation of cisplatin, etoposide, mitomycin, cyclophosphamide, carmustine, and thiotepa was assessed by the Salmonella mutagenicity test after four hours of exposure. Doxorubicin permeation was assessed qualitatively over an eight-hour exposure period using a coloration assay. Saranex-laminated Tyvek was not permeable under the test conditions. Polyethylene-coated Tyvek was slightly permeable to thiotepa and carmustine. Nonporous Tyvek was permeable to all seven drugs, and the Kaycel garment was permeable to all of the drugs except etoposide. In no instance did permeation exceed 3.3% of the applied drug dose. Saranex-laminated Tyvek was the most protective of the barrier garments, followed closely in effectiveness by the polyethylene-coated Tyvek. Clothing made from these two Tyvek composites would allow less air flow and, therefore, would be less comfortable to wear for extended periods. Garments made of nonporous Tyvek or Kaycel would be more comfortable, but their use should be accompanied by an awareness of their potential permeability to certain antineoplastic drugs.

Antineoplastic Agents↗

Comparative permeability of different glioma models to horseradish peroxidase.

Seven different experimental glioma models were studied to determine their capillary permeability to horseradish peroxidase. The models were the autochthonous ASV-viral model, intracerebral (ic) and (sc) injections of rat 9L and RG-2 tumor cell lines, the sc rat S-69C15 tumor cell line, and human glioblastoma cell lines in nude mice. The ASV-viral model showed an average HRP permeability of 63% of tumor volume, the ic RG-2 tumors were 100% permeable, and the ic 9L tumors were 41% permeable. The permeability of the marginal zone (brain-tumor interface) showed similar variation from group to group. In contrast, all of the sc tumors were 100% permeable regardless of the cell line used to create the tumors. Our results show the variability between these glioma models, and suggest that RG-2 ic gliomas and all sc gliomas should be optimal to assess the tumoricidal effect of drugs, because access to the tumor compartment from the vascular compartment is complete.

Animals↗

[Increased erythrocyte membrane permeability to sodium--a risk factor for hypertension].

Monitoring the sodium-lithium counter transport, the authors examined the membrane permeability of red cells for sodium in 70 females and 127 males aged 20 to 70 years. In subjects with normal arterial pressure, the membrane permeability of the red cells showed no age-specific changes or differences in males and females at the age of 20 to 31 years and those over 40 years. Arterial hypertension (AH) according to the WHO criteria was diagnosed in 31% of those studied. The individuals with an increased permeability of the red cell membrane for sodium, exhibited AH twice as frequently as the individuals with the normal or decreased permeability. The rate of AH was found to rise (from 46 to 68%) with age in the subjects with the elevated permeability of the red cell membrane whilst there was no such tendency in the individuals with normal and decreased permeability of the red cell membrane.

Adult↗

Permeability of ovine amnion and amniochorion to urea and water.

Permeability of ovine amnion and amniochorion to urea and water was measured in vitro at various gestational ages. Permeability of amniochorion was between 30 and 100% of that of amnion. Permeability of the membranes to water did not change with fetal age, whereas permeability of amnion to urea decreased from 1.548 +/- 0.541 (49) 10(-4) cm . sec-1 to 0.277 +/- 0.339 (102) 10(-4) cm . sec-1 as fetal age increased. There was a significant relationship between the permeability of the amnion to urea and the concentration of urea in the amniotic fluid. It was concluded that the decrease in membrane permeability to urea may be responsible in part for the increase in amniotic fluid urea concentration observed as gestation progresses.

Amnion↗

The evaluation of the effect of the venous tonic 263-E on capillary permeability in the rabbit after administration by intradermal and intravenous routes.

The effects of 2,5-dihydroxybenzene-1,4-disulphonic acid-bisdiethylamine salt (263-E) and trihydroxyethylrutoside (Vitamin P4) on capillary permeability have been compared in the rabbit. Both drugs were administered by intradermal and intravenous routes. Histamine and bradykinin were injected intradermally to increase permeability. 263-E was sown to be a potent inhibitor of permeability changes induced by histamine and bradykinin intravenous routes. Vitamin P4 maximally inhibited permeability at the lowest intravenous dose level but with increasing dose the inhibition became negligible. After intradernal administration, vitamin P4 did not inhibit permeability but potentiated the permeability response induced by histamine and bradykinin.

Animals↗

The potassium permeability of pancreatic islet cells: mechanisms of control and influence on insulin release.

The K+ permeability of pancreatic islet cells was evaluated by monitoring the efflux of 42K+ or 86Rb+ and measuring the uptake of 42K+ in rat islets. Glucose rapidly and reversibly reduces 42K+ (or 86Rb+) efflux without apparent effect on the rate of 42K+ uptake. This decrease in passive K+ permeability is not secondary to insulin release as it does not require extracellular Ca, and is most marked for changes in the glucose concentration between 3 and 6 mM. This property of glucose is shared by all metabolized sugars, but by none of the nonmetabolized sugars tested. Inhibition of glucose metabolism by islet cells prevents the sugar from decreasing their K+ permeability. Reduction of the redox couple NAD(P)H/NAD(P) appears to be an important link between glucose metabolism and the K+ system in the membrane. A pharmacological decrease of the potassium permeability in islet cells (by tetraethylammonium or 9 aminoacridine) potentiates (and to some extent even mimicks) the insulin-releasing property of glucose. A pharmacological increase (by valinomycin) of this permeability inhibits glucose-stimulated insulin release. The results clearly establish the importance of the decrease in the K+ permeability of B-cell membranes for the physiological response to the stimulation by glucose.

Aminacrine↗

Permeability of the intraocular blood vessels.

This paper reviews quantitative studies on the permeability of the ocular blood vessels. In the retina, where there are tight junctions between the endothelial cells of the microvessels, there is very low permeability even to sodium ions. Glucose is transported through the capillary wall by a carrier, net glucose extraction amounting to about 12 per cent. In the choroid and ciliary processes, where there are fenestrated capillaries, the permeability to plasma proteins is about five times that in the kidney (another tissue with fenestrated capillaries), and ten to thirty times that in heart and skeletal muscle, which has non-fenestrated capillaries without tight junctions. Capillary permeability to low molecular weight substances is high in the choroid; sodium permeability is probably about thirty times that in heart muscle and fifty times that in skeletal muscle. This high permeability results in a high glucose concentration in the tissue fluid enabling rapid glucose movement into the retina.

Animals↗

Increased permeability to sugar following muscle contraction. Inhibitors of protein synthesis prevent reversal of the increase in 3-methylglucose transport rate.

Contractile activity is followed by an increase in the permeability of muscle to sugar. The increase in sugar transport rate persists unchanged for hours before gradually returning to base-line (Holloszy, J. O., and Narahara, H. T. (1965) J. Biol. Chem. 240, 3493-3500). This prolonged delay suggested that the recovery process might involve protein synthesis. We have investigated this possibility. A large increase in permeability to [3-3H]methylglucose occurred in frog sartorius muscles stimulated to contract 60 times/min for 30 min. Cycloheximide or puromycin at concentrations that inhibited protein synthesis completely blocked reversal of the increase in permeability to 3-methylglucose. The recovery process was not irreversibly impaired, as permeability returned to base-line in muscles that were washed and incubated without protein synthesis inhibitors for a second 24-h period. Reversal of the increase in permeability to 3-methylglucose induced by a high concentration of epinephrine was also prevented by cycloheximide. In contrast, cycloheximide did not prevent reversal of the increases in sugar transport rate induced by either insulin or trypsin. These results provide evidence that exercise and catecholamines activate sugar transport by a process that requires protein synthesis for its reversal, while the increases in permeability induced by insulin and trypsin involve a different mechanism.

3-O-Methylglucose↗