THE ACCUMULATION OF ELECTROLYTES : XI. ACCUMULATION OF NITRATE BY VALONIA AND HALICYSTIS

The nitrate concentration in the sap of Valonia macrophysa, Kütz., is at least 2000 times that of the sea water, and in Halicystis Osterhoutii, Blinks and Blinks, at least 500 times that of the sea water.     

THE ACCUMULATION OF ELECTROLYTES : X. ACCUMULATION OF IODINE BY HALICYSTIS AND VALONIA

Analyses of the sap of Halicystis Osterhoutii and of Valonia macrophysa for iodide indicate accumulations of the order of 1000 to 10,000-fold in the first case, and 40 to 250-fold in the second case. The chemical potential of KI, NaI, HI, and CaI₂ is greater inside than outside.     

THE ACCUMULATION OF ELECTROLYTES : II. SUGGESTIONS AS TO THE NATURE OF ACCUMULATION IN VALONIA

It is suggested that K enters chiefly as KOH, whose thermodynamic potential (proportional to the ionic activity product (a _K) (a _OH)) is greater outside than within. As this difference is maintained by the production of acid in the cell K continues to enter, and reaches a greater concentration inside than outside. KOH combines with a weak organic acid which is exchanged for HCl entering from the sea water (or its anion is exchanged for Cl^-), so that KCl accumulates in the sap. Na enters more slowly and its internal concentration remains below that of K. The facts indicate that penetration is chiefly in molecular form. As the system is not in equilibrium the suggestion is not susceptible of thermodynamic proof but it is useful in predicting the behavior of K, Na, and NH₄.     

THE ACCUMULATION OF ELECTROLYTES : III. BEHAVIOR OF SODIUM,POTASSIUM, AND AMMONIUM IN VALONIA

When 0.001 M NH₄Cl is added to sea water containing Valonia macrophysa there seems to be a rapid penetration of undissociated NH₃ (or NH₄OH) which raises the pH value of the sap so that the thermodynamic potential of KOH becomes greater inside than outside and in consequence K leaves the cell: NaOH continues to go in because its thermodynamic potential is greater outside than inside. NH₄Cl accumulates, reaching a much higher concentration inside than outside. This might be explained on the ground that NH₃, after entering, combines with a weak organic acid produced in the cell whose anion is exchanged for the Cl^- of the sea water, or (more probably) the organic acid is exchanged for HCl.     

THE ACCUMULATION OF ELECTROLYTES : I. THE ENTRANCE OF AMMONIA INTO VALONIA MACROPHYSA

When 0.005 M NH₄Cl is added to sea water containing cells of Valonia macrophysa ammonia soon appears in the sap and may reach a concentration inside over 40 times as great as outside. It appears to enter as undissociated NH₃ (or NH₄OH) and tends to reach a pseudoequilibrium in which the activity of undissociated NH₃ (or NH₄OH) is the same inside and outside. When ammonia first enters, the pH value of the sap rapidly rises but it soon reaches a maximum and subsequently falls off. At the same time there is an increase of halide in the sap which, however, does not run a parallel course to the ammonia accumulation, but it comes to a new equilibrium value and remains constant. The increase in NH₃ in the sap is accompanied by a decrease in the concentration of K. As NH₃ enters the specific gravity of the sap decreases and the cells rise to the surface and continue to grow as floating organisms. The growth of the cells is increased.     

THE ACCUMULATION OF ELECTROLYTES : VIII. THE ACCUMULATION OF KCl IN MODELS

Models are described in which KCl enters until its chemical potential becomes much greater inside than outside. The energy needed to accomplish this comes from the chemical reactions occurring in the system and the continual supply of certain materials. An important factor is the maintenance of a lower pH value inside by means of CO₂. This may be analogous to what happens in some living cells. The concentration of K⁺ becomes higher inside, as happens in many living cells, but the concentration of Cl^- does not and in this respect the model differs from many living cells. As in Valonia, potassium tends to go out as KCl when the ionic activity product (K)(Cl) is greater inside but at the same time it tends to enter as KOH since the activity product (K)(OH) is greater outside. The net result is entrance of potassium presumably because the latter process is the more rapid.     

THE ACCUMULATION OF ELECTROLYTES : VII. ORGANIC ELECTROLYTES PART 2

Analyses have been made of the inorganic constituents of the juices expressed from the leaves of Rheum, Rumex, and Oxalis. It has been shown that in all cases there is a large excess of inorganic cations over anions in the sap, the average ratio of cations to anions being 3.8 (Part 1, p. 239). The ash analyses of plant tissues (chiefly leaves) reported in the literature have been examined critically, and it has been shown that the preponderance of inorganic cations over inorganic anions in the ash and in the sap is general. It has been concluded that the excess of inorganic cations is consistent with the view that cations pass into the protoplasm chiefly in the form of hydroxides, and are accumulated either in the form of organic salts (such as the oxalates) or in non-polar linkage. It has been concluded that practically all the potassium and sodium found in plant ash must have been present originally in the form of soluble ionogenic compounds, but that a considerable part of the calcium and magnesium may have been present originally in the form of insoluble salts or as components of non-polar compounds. The methods whereby the cations, particularly potassium, may have been accumulated have been discussed, and it has been concluded that as it does not seem very probable that they enter chiefly as nitrates or bicarbonates we may suppose that they go in to a large extent as hydrates: this is highly probable in the case which has been most carefully investigated (Valonia).     

THE ACCUMULATION OF ELECTROLYTES : VII. ORGANIC ELECTROLYTES PART 1

The inorganic constituents of the sap of Rheum (rhubarb), Rumex (field sorrel), and Oxalis (wood sorrel) show a great preponderance of cations over anions, as would be expected if the cations entered chiefly as hydrates (other possibilities will be discussed in Part 2).     

COMPARISON OF MITOTIC PHENOMENA AND EFFECTS INDUCED BY HYPERTONIC SOLUTIONS IN HELA CELLS

Interphase HeLa cells exposed to solutions that are 1.6 x isotonic manifest a series of morphological transformations, several of which grossly resemble those which occur when untreated cells enter prophase. These include chromosome condensation with preferential localization at the nuclear envelope and nucleolus, ruffling of the nuclear envelope, and polyribosome breakdown. The nucleolus loses its fibrous component and appears diffusely granular. At 2.8 x isotonicity the nuclear envelope is selectively dispersed although other membranes show morphological alterations also. The characteristic transitions of the lysosomes, Golgi complex, and microtubules seen in normal mitosis do not occur during hypertonic treatment. All the changes induced with hypertonic solutions are rapidly reversible, and the nucleus particularly goes through a recovery phase which bears some similarity to that of the telophase nucleus. The prophase-like condensation of the chromatin following exposure of the intact cell to hypertonic medium cannot be reproduced on an ultrastructural level in the isolated nucleus with any known variation in salt concentration, suggesting significant modifications of the nuclear contents during isolation. In addition to these morphological responses, hypertonic solutions also markedly and reversibly depress macromolecular synthesis. The polyribosome disaggregation that results from exposure to hypertonic solutions may be partially prevented by prior exposure to elevated Mg⁺⁺ concentrations; this same ion is also partially effective in preventing the polyribosome breakdown which normally occurs as cells enter mitosis.     

THE ACCUMULATION OF ELECTROLYTES : VI. THE EFFECT OF EXTERNAL pH

It would be natural to suppose that potassium enters Valonia as KCl since it appears in this form in the sap. We find, however, that on this basis we cannot predict the behavior of potassium in any respect. But we can readily do so if we assume that it penetrates chiefly as KOH. We may then say that under normal conditions potassium enters the cell because the ionic activity product (K) (OH) is greater outside than inside. This hypothesis.leads to the following predictions: 1. When the product (K) (OH) becomes greater inside (because the inside concentration of OH^- rises, or the outside concentration of K⁺ or of OH^- falls) potassium should leave the cell, though sodium continues to enter. Previous experiments, and those in this paper, indicate that this is the case. 2. Increasing the pH value of the sea water should increase the rate of entrance of potassium, and vice versa. This appears to be shown by the results described in the present paper. It appears that photosynthesis increases the rate of entrance of potassium by increasing the pH value just outside the protoplasm. In darkness there is little or no growth or absorption of electrolytes. The entrance of potassium by ionic exchange (K⁺ exchanged for H⁺ produced in the cell), the ions passing as such through the protoplasmic surface, does not seem to be important.     

THE ACCUMULATION OF ELECTROLYTES : IV. INTERNAL VERSUS EXTERNAL CONCENTRATIONS OF POTASSIUM

Lowering the potassium in the sea water from 0.011 M to 0.006 M caused an exit of potassium from cells of Valonia macrophysa. Sodium continued to penetrate and the ratio K ÷ Na fell off. The cells ceased to grow but there was no evidence of injury. Increasing the external potassium brought about an increase of the internal concentration of potassium, of halide, of total cations, and of the ratio K ÷ Na inside. These phenomena are to be expected on theoretical grounds.     

THE STRUCTURE OF THE COLLODION MEMBRANE AND ITS ELECTRICAL BEHAVIOR : IX. WATER UPTAKE AND SWELLING OF COLLODION MEMBRANES IN AQUEOUS SOLUTIONS OF ORGANIC ELECTROLYTES AND NON-ELECTROLYTES

1. Dried collodion membranes are known to swell in water and to the same limited extent also in solutions of strong inorganic electrolytes (Carr and Sollner). The present investigation shows that in solutions of organic electrolytes and non-electrolytes, the swelling of dried collodion membranes is not as uniform, but depends on the nature of the solute. 2. The solutions of typically "hydrophilic" substances, e.g., glycerine, glucose, and citric acid, swell collodion membranes only to the same extent as water and solutions of strong electrolytes. In solutions of typically carbophilic substances (e.g., butyric acid, valeric acid, isobutyl alcohol, valeramide, phenol, and m-nitrophenol) the swelling of the membranes is much stronger than in water, according to the concentration used. For the brand of collodion used the swelling in 0.5 M solution was in some cases as high as 26 per cent of the original volume, as compared to 6 to 7 per cent in water. Therefore, in these solutions the "water-wetted dried" collodion membrane is not rigid, inert, and non-swelling, but behaves as a swelling membrane. 3. The solutes which cause an increased swelling of the membranes are accumulated in the latter, the degree of accumulation being markedly parallel with the degree of their specific swelling action. 4. The anomalously high permeabilities of certain carbophilic organic solutes reported by Michaelis, Collander, and Höber find an explanation in the specific interaction of these substances with collodion. 5. The use of the collodion membrane as a model of the ideal porous membrane is restricted to those instances in which no specific interaction occurs between the solute and the collodion.