THE KINETICS OF EXOSMOSIS OF WATER FROM LIVING CELLS

1. The rate of exosmosis of water was studied in unfertilized Arbacia eggs, in order to bring out possible differences between the kinetics of exosmosis and endosmosis. 2. Exosmosis, like endosmosis, is found to follow the equation See PDF for Equation, in which a is the total volume of water that will leave the cell before osmotic equilibrium is attained, x is the volume that has already left the cell at time t, and k is the velocity constant. 3. The velocity constants of the two processes are equal, provided the salt concentration of the medium is the same. 4. The temperature characteristic of exosmosis, as of endomosis, is high. 5. It is concluded that the kinetics of exosmosis and endosmosis of water in these cells are identical, the only difference in the processes being in the direction of the driving force of osmotic pressure

THE EFFECT OF HYDROGEN ION CONCENTRATION ON SWELLING OF CELLS

1. The effect of HCl, NaOH, CO2, and NH3 on the volume of unfertilized Arbacia eggs was tested over a wide range of pH values. 2. No swelling occurred, except in HCl solutions, and there not until after injury or death had occurred. 3. Whereas the volume of erythrocytes and of proteins such as gelatin is known to be dependent on the pH of the solution, such a relation does not exist in the case of living and uninjured cells, at least of the type tested

THE KINETICS OF OSMOTIC SWELLING IN LIVING CELLS

The rate of swelling of unfertilized sea urchin eggs in hypotonic sea water was investigated. Analysis of curves leads to the following conclusions. 1. The rate of swelling follows the equation, See PDF for Equation where Veq., V0, and Vt stand for volume at equilibrium, at first instant, and at time t, respectively, the other symbols having their usual significance. This equation is found to hold over a wide range of temperatures and osmotic pressures. This relation is the one expected in a diffusion process. 2. The rate of swelling is found to have a high temperature coefficient (Q10 = 2 to 3, or µ = 13,000 to 19,000). This deviation from the usual effect of temperature on diffusion processes is thought to be associated with changes in cell permeability to water. The possible influence of changes in viscosity is discussed. 3. The lower the osmotic pressure of the solution, the longer it takes for swelling of the cell. Thus at 15° in 80 per cent sea water, the velocity constant has a value of 0.072, in 20 per cent sea water, of 0.006

THE EFFECT OF VALENCE OF IONS ON CELLULAR PERMEABILITY TO WATER

1. Permeability to water in unfertilized eggs of the sea urchin, Arbacia punctulata, was studied by measuring the rate of swelling in hypotonic dextrose solution. 2. Permeability is greatly affected by addition of electrolytes in low concentration. 3. A decrease in permeability to water was found with increasing valence of the cation, using a series of cobaltammine chlorides in which the valence of the cation ranged from 1 to 6. 4. Conversely, an increase in permeability to water was found with increasing valence of the anion, using two series of potassium salts in which the valence of the anion ranged from 1 to 4, and 1 to 3, respectively. 5. It is concluded that the effect of electrolytes on permeability to water depends chiefly on the sign and the number of charges on the ion, in the sense that positive ions decrease permeability to water, while negative ions increase permeability to water; and the effectiveness of the ion is greater the higher its valence. 6. Antagonism has been demonstrated between cations and anions in their effect on permeability, and the method employed permits quantitative study of such antagonism

THE EFFECT OF SALT CONCENTRATION OF THE MEDIUM ON THE RATE OF OSMOSIS OF WATER THROUGH THE MEMBRANE OF LIVING CELLS

1. Using the unfertilized egg of the sea urchin, Arbacia, as osmometer, it was found that the rate with which water enters or leaves the cell depends on the osmotic pressure of the medium: the velocity constant of the diffusion process is higher when the cell is in concentrated sea water, and lower when the sea water medium is diluted with distilled water. Differences of more than tenfold in the value of the velocity constant were obtained in this way. When velocity constants are plotted against concentration of medium, a sigmoid curve is obtained. 2. These results are believed to indicate that cells are more permeable to water when the osmotic pressure of the medium is high than when it is low. This relation would be accounted for if water should diffuse through pores in a partially hydrated gel, constituting the cell membrane. In a medium of high osmotic pressure, the gel is conceived to give up water, to shrink, and therefore to allow widening of its pores with more ready diffusion of water through them. Conversely, in solutions of lower osmotic pressure, the gel would take up water and its pores become narrow

THE EFFECT OF CERTAIN ELECTROLYTES AND NON-ELECTROLYTES ON PERMEABILITY OF LIVING CELLS TO WATER

1. Permeability to water in unfertilized eggs of the sea urchin, Arbacia punctulata, is found to be greater in hypotonic solutions of dextrose, saccharose and glycocoll than in sea water of the same osmotic pressure. 2. The addition to dextrose solution of small amounts of CaCl2 or MgCl2 restores the permeability approximately to the value obtained in sea water. 3. This effect of CaCl2 and MgCl2 is antagonized by the further addition of NaCl or KCl. 4. It is concluded that the NaCl and KCl tend to increase the permeability of the cell to water, CaCl2 and MgCl2 to decrease it. 5. The method here employed can be used for quantitative study of salt antagonism

THE MECHANISM OF VITAL STAINING WITH BASIC DYES

1. Solutions containing NH4OH and NaOH, and CO2 and HCl may be used to produce various combinations of extracellular and intracellular reactions in starfish eggs, Gonionemus, and Nitella cells. 2. Staining by basic dyes is, with a constant intracellular reaction, favored by increased extracellular alkalinity. With a constant extracellular reaction, staining is hindered by increased intracellular alkalinity. 3. These facts are in opposition to the view that staining of cells by basic dyes is chiefly governed by a combination of the dyes with cell proteins. It is more in harmony with the view that the combination is with a substance or substances of acid nature

THE OSMOTIC PROPERTIES OF LIVING CELLS (EGGS OF ARBACIA PUNCTULATA)

We have attempted to answer the question: How nearly ideal, as an osmometer, is the unfertilized Arbacia egg? The following conclusion have been reached: 1. Volumes can be measured accurately over a wide range of pressures since the cell is in general spherical and does not suffer deformation from its own weight or other factors. 2. The product of volume and pressure is approximately constant, if allowance be made for osmotically inactive cell contents. It is computed that from 7 to 14 per cent of cell volume is occupied by osmotically inactive material. 3. Evidence is presented that no appreciable escape of cell contents occurs while the cell is in hypotonic sea water; that, therefore, the semipermeability of the membrane is approximately perfect, so long as injury to the cell is avoided. 4. In comparison with osmotic pressure the influence of other forces, such as elasticity or surface tension, on cell volume must in these experiments be slight

ON THE MECHANISM OF OPSONIN AND BACTERIOTROPIN ACTION : VI. AGGLUTINATION AND TROPIN ACTION BY PRECIPITIN SERA. CHARACTERIZATION OF THE SENSITIZED SURFACE

As a further test of the theory of tropin action proposed in the preceding paper artificial surfaces have been prepared, and have been found to be phagocytized according to prediction from the theory. Protein was adsorbed on collodion particles according to the technique of F. S. Jones. These particles were then agglutinated and prepared for phagocytosis by the corresponding protein precipitin sera. The precipitating, agglutinating, surface and tropin effects for each serum or serum globulin fraction have been found to be in satisfactory quantitative correspondence. All of these effects were serologically specific; all remained almost unaffected by inactivation of the immune sera for 30 minutes at 56°C. or by washing of the particles after sensitization. The surfaces of particles maximally sensitized by homologous rabbit immune serum or one of its globulin fractions have shown certain characteristic properties, i.e., they were cohesive, had wetting properties characteristic for protein, and were isoelectric at pH. 5.5 to 5.8. The same set of properties were found for immune precipitate in the zone of maximal precipitation. The same properties have also been found for maximally sensitized acid-fast bacteria, and for maximally sensitized sheep erythrocytes. These results indicate, we believe, that precipitation, agglutination, the surface changes and increased phagocytosis are all consequences of one underlying phenomenon. This phenomenon is the specific chemcal combination with, and deposit on the surface of the antigen of antibody protein. The several serological reactions then follow as consequences of the properties of the sensitized surface and of the special environing conditions. The antibody is contained in the globulin fractions of immune serum, and appears to be a globulin with physico-chemical differences from normal serum globulin

ON THE MECHANISM OF OPSONIN AND BACTERIOTROPIN ACTION : II. CORRELATION BETWEEN CHANGES IN BACTERIAL SURFACE PROPERTIES AND IN PHAGOCYTOSIS CAUSED BY NORMAL AND IMMUNE SERA.

The work reported in this and in previous papers (1, 7) demonstrates the following relations for acid-fast bacteria and rabbit polymorphonudear leucocytes: 1. The combination of a substance or substances present in fresh immune rabbit serum, heated or unheated, or in fresh unheated normal rabbit serum, with a substance or substances in the bacterial surface causes an increase in cohesiveness, decrease in surface potential difference and characteristic alteration in wetting properties of the bacteria, and prepares the bacteria for phagocytosis. 2. (a) The effective substance or substances in the serum may become so altered as the result of heating or aging that combination with the bacterial surface, while causing changes in bacterial surface properties indistinguishable by the present physical-chemical tests from these just mentioned, may not lead to phagocytosis, or may lead to phagocytosis with a prezone not paralleled by a prezone in the changes in surface properties. (b) Sensitization of bacteria with human sera causes changes in surface properties similar to those caused by rabbit sera, but does not lead to phagocytosis by rabbit leucocytes. The spreading requirements of rabbit polymorphonuclear leucocytes are evidently highly selective