Search PubMed⌕ Search

Biomedical subjects

J Gutknecht

Publications and source records attributed to J Gutknecht.

52 records · Page 3Linked to original sources
PubMed1978

Electrically silent anion transport through lipid bilayer membranes containing a long-chain secondary amine.

The permeability properties of planar lipid bilayers made from egg lecithin, n-decane and a long-chain secondary amine (n-lauryl [trialkylmethyl]amine) are described. Membranes containing the secondary amine show halide selectivity and high conductance at pH less than 6, as estimated by measurements of zero-current potentials generated by NaBr activity gradients. In the absence of halide ions, the membranes show H+ selectivity, although the total membrane conductance is relatively low. In 0.1 M NaBr both the membrane conductance (Gm) and the Br- self-exchange flux (JBr) are proportional to H+ concentration over the pH range of 7 to 4, and both JBr and Gm saturate at pH less than 4. However, JBr is always more than 100 times the flux predicted from Gm and the transference number for Br-. Thus, greater than 99% of the observed (tracer) flux is electrically silent and is not a Br2 or HBrO flux because the reducing agent, S2O3=, has no effect on JBr. At pH 7, JBr is proportional to Br- concentration over the range of 1-340 mM, with no sign of saturation kinetics. Both urea and sulfate tracer permeabilities are low and are unaffected by pH. The results can be explained by a model in which the secondary amine behaves as a monovalent, titratable carrier which exists in three chemical forms (C, CH+, and CHBr). Br- crosses the membrane primarily as the neurtal complex (CHBr). The positively charged carrier (CH+) crosses the membrane slowly compared to CHBr, but CH+ is the principal charge carrier in the membrane. At neurtal pH greater than 99% of the amine is in the nonfunctional form (C), which can be converted to CH+ or CHBr by increasing the H+ or Br- concentrations. The permeability properties of these lipid bilayers resemble in many respects the permeability properties of red cell membranes.

Amines↗
PubMed1977

Diffusion of carbon dioxide through lipid bilayer membranes: effects of carbonic anhydrase, bicarbonate, and unstirred layers.

Diffusion of (14)C-labeled CO(2) was measured through lipid bilayer membranes composed of egg lecithin and cholesterol (1:1 mol ratio) dissolved in n-decane. The results indicate that CO(2), but not HCO(3-), crosses the membrane and that different steps in the transport process are rate limiting under different conditions. In one series of experiments we studied one-way fluxes between identical solutions at constant pCO(2) but differing [HCO(3-)] and pH. In the absence of carbonic anhydrase (CA) the diffusion of CO(2) through the aqueous unstirred layers is rate limiting because the uncatalyzed hydration-dehydration of CO(2) is too slow to permit the high [HCO(3-)] to facilitate tracer diffusion through the unstirred layers. Addition of CA (ca. 1 mg/ml) to both bathing solutions causes a 10-100-fold stimulation of the CO(2) flux, which is proportional to [HCO(3-)] over the pH range 7-8. In the presence of CA the hydration- dehydration reaction is so fast that CO(2) transport across the entire system is rate limited by diffusion of HCO(3-) through unstirred layers. However, in the presence of CA when the ratio [HCO(3-) + CO(3=)]:[CO(2)] more than 1,000 (pH 9-10) the CO(2) flux reaches a maximum value. Under these conditions the diffusion of CO(2) through the membrane becomes rate limiting, which allows us to estimate a permeability coefficient of the membrane to CO(2) of 0.35 cm s(-1). In a second series of experiments we studied the effects of CA and buffer concentration on the net flux of CO(2). CA stimulates the net CO(2) flux in well buffered, but no in unbuffered, solutions. The buffer provides a proton source on the upstream side of the membrane and proton sink on the downstream side, thus allowing HCO(3-) to facilitate the net transport of CO(2) through the unstirred layers.

Bicarbonates↗
PubMed1976

Ion transport studies and determination of the cell wall elastic modulus in the marine alga Halicystis parvula.

Using cultured cells of the marine alga, Halicystis parvula, we measured the concentrations of 11 inorganic ions in the vacuolar sap and the electrical potential difference (PD) between the vacuole and the external solution. In normal cells under steady-state conditions a comparison of the electrochemical equilibrium (Nernst) potential for each ion with the PD of -82 mV (inside negative) indicates that Na+ and K+ are actively transported out of the vacuole whereas all anions are pumped into the cell. Although the [K+] in the vacuole is only 9 mM, the cytoplasmic [K+] is about 420 mM, which suggests that the outwardly directed pump is at the tonoplast. Using large Halicystis cells we perfused the vacuole with an artificial seawater and conducted a short-circuit analysis of ion transport. The short-circuit current (SCC) of 299 peq - cm-2-s-1 is not significantly different from the net influx of Cl-. There is a small, but statistically significant net efflux of K+ (less than 1 pmol-cm-2.-1), while the influx and efflux of Na+ are not significantly different. Therefore, the SCC is a good measure of the activity of the Cl- pump. Finally, we measured the volumetric elastic modulus (epsilon) of the cell wall by measuring the change in cell volume when the internal hydrostatic pressure was altered. The value of epsilon at applied pressures between 0 and 0.4 atm is about 0.6 atm, which is at least 100-fold lower than the values of epsilon for all other algae which have been studied.

Biological Transport, Active↗
PubMed1975

Osmotic regulation in the marine alga, Codium decorticatum. I. Regulation of turgor pressure by control of ionic composition.

Codium decorticatum regulates its internal ionic composition and osmotic pressure in response to changes in external salinity. Over a salinity range of 23 to 37% (675 to 1120 mosmol/kg) Codium maintains a constant turgor pressure of 95 mosmol/kg (2.3 atm), observed as a constant difference between internal and external osmotic pressures. The changes in internal osmotic pressure are due to changes in intracellular inorganic ions. At 30 0/00 salinity the major intracellular ions are present in the following concentrations (mmol/kg cell H20): K+, 295; Na+, 255; Cl-, 450. At different salinities intracellular ion concentrations remain in constant proportion to the external ion concentrations, and thus the equilibrium potentials are approximately constant. The potential difference between the vacuole and seawater (-76 mV), whici is predominantly a K+ diffusion potential, is also constant with changing salinity. Comparison of the equilibrium potentials with the vacuole potential suggests that Cl- is actively absorbed and Na+ actively extruded, whereas K+ may be passively distributed between the vacuole and seawater. Turgor pressure does not change with environmental hydrostatic pressure, and increasing the external osmotic pressure with raffinose elicits a response similar to that obtained by increasing the salinity. These two results suggest that the stimulus for turgor regulation is a change in turgor pressure rather than a change in internal hydrostatic pressure or ion concentrations.

Chlorophyta↗
PubMed1973

Diffusion of weak acids across lipid bilayer membranes: effects of chemical reactions in the unstirred layers.

Chemical reactions in the aqueous unstirred layers of solution adjacent to a membrane can have dramatic effects on the diffusion of solutes across that membrane. This is demonstrated by the diffusion of labeled salicylate and salicylic acid across a phospholipid bilayer membrane. Two types of chemical reactions are considered. The first is an isotopic exchange reaction between the ionic and nonionic forms of a weak acid, HA + *A(-) [unknown] H*A + A(-). This reaction provides a way of estimating the true membrane permeability to a highly permeant weak acid and also a way of estimating the thickness of the unstirred layers. The second chemical reaction, the dissociation of a weak acid, HA [unknown] H(+) + A(-), can be used to show how the presence or absence of buffers in the unstirred layers controls the net transport of permeant weak acids across a membrane. In principle, the addition of appropriate "antacid" buffers to salicylates can increase their rate of absorption from the stomach.

Carbon Radioisotopes↗
PubMed1972

The permeability of thin lipid membranes to bromide and bromine.

Thin lipid (optically black) membranes were made from sheep red cell lipids dissolved in n-decane. The flux of Br across these membranes was measured by the use of tracer (82)Br. The unidirectional flux of Br (in 50-100 mM NaBr) was 1-3 x 10(-12) mole/cm(2)sec. This flux is more than 1000 times the flux predicted from the membrane electrical resistance (>10(8) ohm-cm(2)) and the transference number for Br(-) (0.2-0.3), which was estimated from measurements of the zero current potential difference. The Br flux was not affected by changes in the potential difference imposed across the membrane (+/-60 mv) or by the ionic strength of the bathing solutions. However, the addition of a reducing agent, sodium thiosulfate (10(-3)M), to the NaBr solution bathing the membrane caused a 90% reduction in the Br flux. The inhibiting effect of S(2)O(3) (=) suggests that the Br flux is due chiefly to traces of Br(2) in NaBr solutions. As expected, the addition of Br(2) to the NaBr solutions greatly stimulated the Br flux. However, at constant Br(2) concentration, the Br flux was also stimulated by increasing the Br(-) concentration, in spite of the fact that the membrane was virtually impermeable to Br(-). Finally, the Br flux appeared to saturate at high Br(2) concentrations, and the saturation value was roughly proportional to the Br(-) concentration. These results can be explained by a model which assumes that Br crosses the membrane only as Br(2) but that rapid equilibration of Br between Br(2) and Br(-) occurs in the unstirred layers of aqueous solution bathing the two sides of the membrane. A consequence of the model is that Br(-) "facilitates" the diffusion of Br across the unstirred layers.

Animals↗
PubMed1970

Ionic peremability of thin lipid membranes. Effects of n-alkyl alcohols, polyvalent cations, and a secondary amine.

Ultrathin (black) lipid membranes were made from sheep red cell lipids dissolved in n-decane. The presence of aliphatic alcohols in the aqueous solutions bathing these membranes produced reversible changes in the ionic permeability, but not the osomotic permeability. Heptanol (8 mM), for example, caused the membrane resistance (R(m)) to decrease from >10(8) to about 10(5) ohm-cm(2) and caused a marked increase in the permeability to cations, especially potassium. In terms of ionic transference numbers, deduced from measurements of the membrane potential at zero current, T(cat)/T(Cl) increased from about 6 to 21 and T(K)/T(Na) increased from about 3 to 21. The addition of long-chain (C(8)ndash;C(10)) alcohols to the lipid solutions from which membranes were made produced similar effects on the ionic permeability. A plot of log R(m) vs. log alcohol concentration was linear over the range of maximum change in R(m), and the slope was -3 to -5 for C(2) through C(7) alcohols, suggesting that a complex of several alcohol molecules is responsible for the increase in ionic permeability. Membrane permselectivity changed from cationic to anionic when thorium or ferric iron (10(-4)M) was present in the aqueous phase or when a secondary amine (Amberlite LA-2) was added to the lipid solutions from which membranes were made. When membranes containing the secondary amine were exposed to heptanol, R(m) became very low (10(3)-10(4) ohm-cm(2)) and the membranes became perfectly anion-selective, developing chloride diffusion potentials up to 150 mv.

Alcohols↗
PubMed1968

Salt transport in Valonia: inhibition of potassium uptake by small hydrostatic pressures.

Internally perfused cells of the marine alga Valonia actively transport potassium from external seawater into the cell vacuole. This active uptake of potassium is reduced by restoring the normal turgor pressure of 1 atmosphere by means of a mercury manometer attached to the internal perfusion system. This inhibition of salt uptake by a small hydrostatic pressure suggests that Valonia and other walled cells may regulate their turgor pressures by adjusting their rates of salt uptake.

Biological Transport, Active↗
PubMed1967

Membranes of Valonia ventricosa: apparent absence of water-filled pores.

Osmotic and diffusional permeabilities to water have been measured in internally perfused cells of Valonia ventricosa. The osmotic and diffusional permeability coefficients for the protoplast are identical, 2.4 x 10(-4) centimeter per second. Thus, both osmotic and diffusional flows can occur by the same mechanism, that is, by diffusion; and there is no need to postulate the existence of water-filled pores in the membranes of this cell. Supporting evidence for this conclusion is the absence of solvent-solute interactions, that is, "solvent drag," for water, urea, and methanol crossing the protoplast of Valonia.

Cell Wall↗
PubMed1967

Ion fluxes and short-circuit current in internally perfused cells of Valonia ventricosa.

Ion transport in the giant celled marine alga, Valonia ventricosa, was studied during internal perfusion and short-circuiting of the vacuole potential. The perfusing and bathing solutions were similar to natural Valonia sap and contained the following concentrations of major ions: Na 51, K 618, and Cl 652 mM. The average short-circuit current (I(sc)) was 97 pEq/cm(2) sec (inward positive current), and the average open-circuit potential difference (PD) was 74 mv (vacuole positive to external solution). Perfused and short-circuited cells showed a small net influx of Na (2.0 pEq/cm(2) sec) and large net influxes of K (80 pEq/cm(2) sec) and Cl (50 pEq/cm(2) sec). Unidirectional K influx was proportional to I(sc), but more than one-half of the I(sc) remained unaccounted for. Both the I(sc) and PD were partly light-dependent, declining rapidly during the first 1-2 min of darkness. Ouabain (5 x 10(-4)M) had little effect on the influx of Na or K and had no effect on I(inf) or PD. Fluid was absorbed at a rate of about 93 pliter/cm(2) sec. Reversing the direction of fluid movement by adding mannitol to the outside solution had little effect on ion movements. The ionic and electrical properties of normal and perfused cells of Valonia are compared.

Chlorides↗