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Membrane potentials in the Donnan equilibrium. II.

Measurements were made of electromotive force in the Donnan equilibrium of systems containing dilute solutions of protein and acid. Removal of the membrane produced a decrease of no more than 2 to 4 mv. in electromotive force, while the membrane potentials, as estimated by the usual arbitrary assumption, were of the order of 12 to 34 mv. Ion ratios, as calculated from analyses for total chloride, were definitely greater than those calculated from the electromotive force of cells with salt bridges, as if there had been combination of some of the chloride ion with protein.

Chlorides↗

The freezing point depression of mammalian tissues in relation to the question of osmotic activity of cell fluid.

The freezing point depression of freshly excised frozen tissues, pulverized in a hydraulic press or in a mortar, is greater than that of plasma. Even at 0 degrees C. the freezing point depression of such homogenates increases significantly with time. Dilution data indicate that such freezing point data are valid. The presence of intact cells has been shown in smears of tissues pulverized in a mortar, but not in smears of those crushed in a hydraulic press. The osmolarity of various diluent solutions affects the calculated osmotic activity of tissue homogenates presumably because of delayed diffusion between the diluent and cell fluid. With a hypertonic NaCl diluent, spuriously low values of tissue osmotic activity are found from calculations assuming instantaneous mixing between homogenates and diluents. The limitations of data from cryoscopic experiments and from tissue-swelling experiments are discussed in relation to the basic question of whether or not cell fluid is isotonic to extracellular fluid.

Animals↗

The contributions of normal and anomalous osmosis to the osmotic effects arising across charged membranes with solutions of electrolytes.

The osmotic effect arising across a porous membrane separating the solution of an electrolyte from water (or a more dilute solution) is ordinarily due to both normal osmosis, as it occurs also with non-electrolytes, and to "anomalous" osmosis. It is shown that the normal osmotic component cannot be measured quantitatively by the conventional comparison with a non-electrolytic reference solute. Anomalous osmosis does not occur with electroneutral membranes. Accordingly, with membranes which can be charged and discharged reversibly (without changes in geometrical structure), such as many proteinized membranes, the osmotic effects caused by an electrolyte can be measured both when only normal osmosis arises (with the membrane in the electroneutral state) and when normal as well as anomalous osmosis occurs (with the membrane in a charged state). The difference between these two effects is the true anomalous osmosis. Data are presented on the osmotic effects across an oxyhemoglobin membrane in the uncharged state at pH 6.75 and in two charged states, positive at pH 4.0 and negative at pH 10.0, with solutions of a variety of electrolytes using a concentration ratio of 2:1 over a wide range of concentrations. The rates of the movement of liquid across the membrane against an inconsequentially small hydrostatic head are recorded instead of, as conventional, the physiologically less significant pressure rises after a standard time.

Electrolytes↗

Electrokinetic membrane processes in relation to properties of excitable tissues. I. Experiments on oscillatory transport phenomena in artificial membranes.

An artificial system is studied consisting of salt solutions of different concentrations separated by a porous, "charged" membrane, through which a constant electric current is passed. Experiments on such systems demonstrate rhythmic variations of the transmembrane potential and the membrane resistance, which are concomitant with an oscillatory streaming of water solution across the membrane. The repetitive oscillations can be of a damped or undamped type dependent on the "stimulating" current density. A qualitative discussion of the mechanism of the oscillations is given. It centers around the periodic resistance changes in the membrane, which result from a complicated interplay between the driving forces present. The importance of electro-osmotic effects is emphasized. A few comparisons relating to possible electrophysiological implications are presented. In the metastable state of this membrane oscillator, "make" and "break" responses can be triggered by electric as well as by mechanical (pressure) "stimuli."

Biological Transport↗

The potential of water in mammalian tissues.

Melting point depression was used as an index of the water potential of rat tissues and serum. Organs removed from anesthetized rats were immediately frozen in liquid nitrogen and ground with mortar and pestle. Aliquots of the resulting frozen powder were suspended in chilled liquid silicone. While the suspension was vigorously stirred and warmed at a constant rate, the temperature of the melting mixture was measured. The melting curves of rat muscle, liver, heart, and brain were not significantly different from those of rat serum. The melting curve depression of whole kidney was greater than that of serum; this was demonstrated to be due to hypertonicity of the renal medullary area alone. It was demonstrated that autolysis will rapidly increase the depression of the melting curve of tissue. It is concluded that within the limits of the method used the melting point depression, and hence the water potential, of intracellular and extracellular fluids is the same.

Animals↗