THE STRUCTURE OF THE COLLODION MEMBRANE AND ITS ELECTRICAL BEHAVIOR : X. AN EXPERIMENTAL TEST OF SOME ASPECTS OF THE TEORELL AND MEYER-SIEVERS THEORIES OF ELECTRICAL MEMBRANE BEHAVIOR

1. The Teorell, Meyer-Sievers theory characterizes the electrochemical behavior of membranes by their selectivity constant "A_p" which is derived conventionally from concentration potential measurements at various concentration levels. The selectivity constant may, however, be derived also from entirely independent, different experimental data, namely base exchange studies. The constants arrived at in this second way are designated as "A_b." The selectivity constants derived by these two methods must be in reasonable, at least semiquantitative agreement if the basic assumptions of the theory are correct. 2. The selectivity constants A_p and A_b were determined for eleven different sets of membranes of different electrochemical activity and of different (8.2 to 80 volume per cent) water content. 3. The potentiometric selectivity constants A_p are in most cases several orders of magnitude greater than the corresponding A_b values. With membranes of great porosity and high electrochemical activity the A_b values approach at least in order of magnitude the A_p values. 4. It is concluded that the unexpectedly large discrepancy between the A_p and A_b values is due to some inherent weakness of the Teorell, Meyer-Sievers theory, most likely to its neglect of any structural factors.     

THE ORIENTATION OF ANIMALS BY OPPOSED BEAMS OF LIGHT

When orientation is attained under the influence of beams of parallel light opposed at 180° the deflection θ from a path at right angles to the beams is given by tan See PDF for Equation, where I ₁ and I ₂ are the photic intensities and H is the average angle between the photoreceptive surfaces. This expression is independent of the units in which I is measured, and holds whether the primary photosensory effect is proportional to I or to log I. When photokinetic side-to-side motions of the head occur, H decreases with increasing total acting light intensity, but increases if higher total light intensity restricts the amplitude of random movements; in each case, H is very nearly proportional to log I ₁ I ₂. For beams of light at 90°, See PDF for Equation. The application of these equations to some particular instances is discussed, and it is shown why certain simpler empirical formulæ previously found by others yield fair concordance with the experimental data. The result is thus in complete accord with the tropism theory, since the equations are based simply on the assumption that when orientation is attained photic excitation is the same on the two sides.     

THE KINETICS OF PENETRATION : I. EQUATIONS FOR THE ENTRANCE OF ELECTROLYTES

When the only solute present is a weak acid, HA, which penetrates as molecules only into a living cell according to a curve of the first order and eventually reaches a true equilibrium we may regard the rate of increase of molecules inside as See PDF for Equation where P_M is the permeability of the protoplasm to molecules, M_o, denotes the external and M_i the internal concentration of molecules, A_i denotes the internal concentration of the anion A^- and See PDF for Equation (It is assumed that the activity coefficients equal 1.) Putting P_MF_M = V_M, the apparent velocity constant of the process, we have See PDF for Equation where e denotes the concentration at equilibrium. Then See PDF for Equation where t is time. The corresponding equation when ions alone enter is See PDF for Equation. where K is the dissociation constant of HA, P_A is the permeability of the protoplasm to the ion pair H⁺ + A^-, and A_ie denotes the internal concentration of A_i at equilibrium. Putting P_AKF_M = V_A, the apparent velocity constant of the process, we have See PDF for Equation and See PDF for Equation When both ions and molecules of HA enter together we have See PDF for Equation where S_i = M_i + A_i and S_ie is the value of S_i at equilibrium. Then See PDF for Equation V_M, V_A, and V_MA depend on F_M and hence on the internal pH value but are independent of the external pH value except as it affects the internal pH value. When the ion pair Na⁺ + A^- penetrates and Na_i = BA_i, we have See PDF for Equation and See PDF for Equation where P _NaA is the permeability of the protoplasm to the ion pair Na⁺ + A^-, Na_o and Na_i are the external and internal concentrations of Na⁺, See PDF for Equation, and V _Na is the apparent velocity constant of the process. Equations are also given for the penetration of: (1) molecules of HA and the ion pair Na⁺ + A^-, (2) the ion pairs H⁺ + A^- and Na⁺ + A^-, (3) molecules of HA and the ion pairs Na⁺ + A^- and H⁺ + A^-. (4) The penetration of molecules of HA together with those of a weak base ZOH. (5) Exchange of ions of the same sign. When a weak electrolyte HA is the only solute present we cannot decide whether molecules alone or molecules and ions enter by comparing the velocity constants at different pH values, since in both cases they will behave alike, remaining constant if F_M is constant and falling off with increase of external pH value if F_M falls off. But if a salt (e.g., NaA) is the only substance penetrating the velocity constant will increase with increase of external pH value: if molecules of HA and the ions of a salt NaA. penetrate together the velocity constant may increase or decrease while the internal pH value rises. The initial rate See PDF for Equation (i.e., the rate when M_i = 0 and A_i = 0) falls off with increase of external pH value if HA alone is present and penetrates as molecules or as ions (or in both forms). But if a salt (e.g., NaA) penetrates the initial rate may in some cases decrease and then increase as the external pH value increases. At equilibrium the value of M_i equals that of M_o (no matter whether molecules alone penetrate, or ions alone, or both together). If the total external concentration (S_o = M_o + A_o) be kept constant a decrease in the external pH value will increase the value of M_o and make a corresponding increase in the rate of entrance and in the value at equilibrium no matter whether molecules alone penetrate, or ions alone, or both together. What is here said of weak acids holds with suitable modifications for weak bases and for amphoteric electrolytes and may also be applied to strong electrolytes.     

THE KINETICS OF STARVATION

1. The relative weight loss of pigeons on starving rations may be represented by a simple modification of the law of autocatalysis. 2. An equation giving the actual weight of the animals during the 53 day period of the experiment is derived by means of the hypothesis that the process of inanition is governed by the simultaneous velocity relations between two independent, homogeneous systems, such that M_T = (A ₀ → 0) + (B ₀ → B_C) in which A is taken proportional to the amount of carbohydrate-fat reserves, and B proportional to the amount of reacting body protoplasm. 3. The course of starvation is governed by the rate of destruction of body protein, but it is modified by the amount and by the rate of destruction of reserve materials. 4. The processes of breaking down tissues are analogous to those by which tissues are synthesized. 5. The close agreement between observed and calculated values suggests that the original assumptions set forth in deriving the foregoing equations were valid. 6. Attention is called to the fact that the time of onset of beriberi in pigeons is coincident with the half period of starvation.     

THEORY AND MEASUREMENT OF VISUAL MECHANISMS : X. MODIFICATIONS OF THE FLICKER RESPONSE CONTOUR,AND THE SIGNIFICANCE OF THE AVIAN PECTEN

1. When there is projected on the retina (man, monocularly) the shadow of a grid which forms a visual field in several distinct pieces (not including the fovea in the present tests), the ordinary properties of the flicker recognition contour (F vs. log I) as a function of the light-time cycle fraction (t_L) can be markedly disturbed. In the present experiments flicker was produced by the rotation of a cylinder with opaque vertical stripes. In the absence of such a grid shadow the "cone" segments of the contours form a set in which F_max. and the abscissa of inflection are opposite but rectilinear functions of t_L, while the third parameter of the probability integral (σ''_{log I}) remains constant. This is the case also with diverse other animals tested. In the data with the grid, however, analysis shows that even for low values of t_L (up to 0.50) there occurs an enhancement of the production of elements of neural effect, so that F_max. rises rather than falls as ordinarily with increase of t_L, although σ''_{log I} stays constant and hence the total number of acting units is presumed not to change. This constitutes valid evidence for neural integration of effects due to the illumination of separated retinal patches. Beginning at t_L = 0.75, and at 0.90, the slope of the "cone" curve is sharply increased, and the maximum F is far above its position in the absence of the grid. The decrease of σ''_{log I} (the slope constant) signifies, in terms of other information, an increase in the number of acting cone units. The abscissa of inflection is also much lowered, relatively, whereas without the grid it increases as t_L is made larger. These effects correspond subjectively to the fact that at the end-point flicker is most pronounced, on the "cone" curve, along the edges of the grid shadow where contrast is particularly evident with the longer light-times. The "rod" portion of the F - log I contour is not specifically affected by the presence of the grid shadow. Its form is obtainable at t_L = 0.90 free from the influence of summating "cone" contributions, because then almost no overlapping occurs. Analysis shows that when overlapping does occur a certain number of rod units are inhibited by concurrent cone excitation, and that the mean contribution of elements of neural action from each of the non-inhibited units is also reduced to an extent depending on the degree of overlap. The isolated "rod" curve at t_L = 0.90 is quite accurately in the form of a probability integral. The data thus give a new experimental proof of the occurrence of two distinct but interlocking populations of visual effects, and experimentally justify the analytical procedures which have been used to separate them. 2. The changing form of the F - log I contour as a function of t_L, produced in man when the illuminated field is divided into parts by a shadow pattern, is normally found with the bird Taeniopygia castenotis (Gould), the zebra finch. The retina has elements of one general structural type (cones), and the F - log I contour is a simplex symmetrical probability integral. The eye of this bird has a large, complex, and darkly pigmented pecten, which casts a foliated shadow on the retina. The change in form of the F - log I curve occurs with t_L above 0,50, and at t_L = 0.90 is quite extreme. It is more pronounced than the one that is secured in the human data with the particular grid we have used, but there is no doubt that it could be mimicked completely by the use of other grids. The increase of flicker acuity due to the pecten shadow is considerable, when the dark spaces are brief relative to the light. The evidence thus confirms the suggestion (Menner) drawn from comparative natural history that the visual significance of the avian pecten might be to increase the sensory effect of small moving images. It is theoretically important that (as in the human experiment) this may be brought about by an actual decrease of effective retinal area illuminated. It is also significant theoretically that despite the presence of shadows of pecten or of grid, and of the sensory influences thus introduced, the probability integral formulation remains effective.     

THE OXIDATION-REDUCTION POTENTIAL OF THE LUCIFERIN-OXYLUCIFERIN SYSTEM

The oxidation-reduction potential of the Cypridina luciferin-oxyluciferin system determined by a method of "bracketing" lies somewhere between that of anthraquinone 2-6-di Na sulfonate (E_o ^'' at pH of 7.7 = –.22) which reduces luciferin, and quinhydrone (E_o ^'' at pH of 7.7 = +.24), which oxidizes luciferin. Systems having an E_o ^'' value between –.22 and +.24 volt neither reduce oxyluciferin nor oxidize luciferin. If the luciferin-oxyluciferin system were truly reversible considerable reduction and oxidation should occur between –.22 and +.24. The system appears to be an irreversible one, with both "apparent oxidation" and "apparent reduction potentials" in Conant''s sense. Hydrosulfites, sulfides, CrCl₂, TiCl₃, and nascent hydrogen reduce oxyluciferin readily in absence of oxygen but without luminescence. Luminescence only appears in water solution if luciferin is oxidized by dissolved oxygen in presence of luciferase. Rapid oxidation of luciferin by oxygen without luciferase or oxidation by K₃Fe(CN)₆ in presence of luciferase but without oxygen never gives luminescence.     

STUDIES ON THE METABOLISM OF AUTOTROPHIC BACTERIA : II. THE NATURE OF THE CHEMOSYNTHETIC REACTION

In a study of chemosynthesis (the fixation of CO₂ by autotrophic bacteria in the dark) in Thiobacillus thiooxidans, the data obtained support the following conclusions: 1. CO₂ can be fixed by "resting cells" of Thiobacillus thiooxidans; the fixation is not "growth bound." 2. The physiological condition of the cell is of considerable importance in determining CO₂ fixation. 3. CO₂ fixation can occur in the absence of oxidizable sulfur in "young" cells. The extent of this fixation appears to be dependent upon the pCO₂. 4. CO₂ fixation can also occur under anaerobic conditions and the presence of sulfur does not influence such fixation. 5. However, in the CO₂ fixation by cells in the absence of sulfur, only a limited amount of CO₂ can be fixed. This amount is approximately 40 µl. CO₂ per 100 micrograms bacterial nitrogen. After a culture has utilized this amount of CO₂ it no longer has the ability to fix CO₂ but releases it during its respiration. 6. Relatively short periods of sulfur oxidation can restore the ability of cells to fix CO₂ under conditions where sulfur oxidation is prevented. 7. It is possible to oxidize sulfur in the absence of CO₂ and to store the energy thus formed within the cell. It is then possible to use this energy at a later time for the fixation of CO₂ in the entire absence of sulfur oxidation. 8. Cultures of Thiobacillus thiooxidans respiring on sulfur utilize CO₂ in a reaction which proceeds to a zero concentration of CO₂ in the atmosphere. 9. CO₂ may act as an oxidizing agent for sulfur. 10. Hydrogen is not utilized by the organism. 11. It is possible to selectively inhibit sulfur oxidation and CO₂ fixation.     

THE INACTIVATION OF BACTERIOPHAGE BY MERCURY BICHLORIDE; THE REACTIVATION OF BICHLORIDE-INACTIVATED PHAGE

1. The inactivation of antistaphylococcus bacteriophage suspended in infusion broth at pH 7.6 and 22°C. by HgCl₂ proceeds according to the equation dP/dt = k [HgCl₂] [P_o – P_i] over the range studied. 2. This inactivation can be reversed by precipitation of Hg⁺⁺ with H₂S. In the present experiments the inactivation was carried out until only some 5 per cent of the initial phage remained active. After reactivation the [P] had increased to 100 per cent of the initial [P].     

THE EFFECT OF CARBON DIOXIDE TENSION ON THE METABOLISM OF CEREBRAL CORTEX AND MEDULLA OBLONGATA

Manometric measurements were made of oxygen uptake (Q _OO2) and aerobic lactic acid output (Q_G) by slices of cerebral cortex and medulla oblongata of the cat in the presence of mixtures of 1, 5, and 20 volumes per cent of carbon dioxide in oxygen. The concentrations of NaHCO₃ and NaCl in the medium were varied to maintain constant pH and sodium ion concentrations. The calcium ion concentration was 0.0002 M. At pH 7.5 under these conditions, an increase in carbon dioxide from 1 per cent to 5 per cent doubled the Q_G of both tissues but did not alter Q _OO2; an increase from 5 per cent to 20 per cent carbon dioxide had no further effect on Q_G in either tissue or Q _OO2 of cortex, but did depress the Q _OO2 of medulla. At pH 8.1, an increase in carbon dioxide from 1 per cent to 5 per cent raised the Q _OO2 and Q_G of cortex by about 60 per cent. Measurements at low oxygen tension carried out previously in phosphate medium were repeated in bicarbonate medium to obtain data for the combined output of lactic acid and carbon dioxide (Q_A). When the oxygen in the gas phase was decreased from 95 to 3 volumes per cent, the lactic acid output as measured colorimetrically increased by 114 mg./gm. in cortex and by 8 mg./gm. in medulla; Q_A increased from 12.3 to 13.5 in cortex and decreased from 5.1 to 3.8 in medulla.     

RELATION OF OXYGEN TENSION AND TEMPERATURE TO THE TIME OF REDUCTION OF CYTOCHROME

The time for the appearance of the cytochrome C absorption band after shaking a suspension of bakers'' yeast with various O₂-N₂ mixtures was determined at each of six temperatures. At each temperature a linear relation between this interval—called the reduction time—and O₂ tension was found. It was shown: 1. That under our experimental conditions, absorption bands of cytochrome were seen when the O₂ tension of the suspension was reduced to, or below, a certain pressure which was found to be specific for each temperature (this pressure is provisionally considered to be identical with or very near to the "critical O₂ tension" usually found in Q _OO2-O₂-tension relationships); 2. That the x-axis intercept obtained from the reduction time - O₂-tension plot gives the value of the "critical" O₂ pressure at each temperature; 3. That the O₂ tension within the suspension is reduced by the respiratory activity of the yeast cells. An equation describing these observations is given and is used in calculating rates of O₂ consumption from measurements of reduction time of cytochrome. The average difference between the calculated values and the manometric measurements of Q _OO2 was found to be 6.6 per cent. A rapid optical method of measuring rates of O₂ consumption based on the findings of these experiments is proposed for use with cytochrome-containing microorganisms.     

THE RHEOLOGY OF THE BLOOD. III

The authors have confirmed the fact that blood serum and plasma behave rheologically like a true viscous liquid. It is true for whole blood only to a first approximation, but with this reservation they have studied the available data and extended the equation of Bingham and Durham to cover protein solutions of various concentrations and at various temperatures as well as mixtures of proteins and corpuscles present in whole blood. If Φ is the fluidity of whole blood, Φ₁ is the fluidity of water and ΔΦ = Φ – Φ₁, then ΔΦ = β₁ b ₁ + β₂ b ₂ + β₃ b ₃ + ··· where β₁, β₂, β₃, etc., are constants for the fluidity lowering of the salts, albumin, globulin, fibrinogen, and the corpuscles, etc., present in the whole blood. The conclusions from the data referred to are intended to buttress this simple equation (6).     

THE EFFECT OF CYSTINE AND GLYCOCOLL ON THE OXIDATION OF SODIUM LACTATE BY HYDROGEN PEROXIDE

The rate of production of CO₂ from sodium lactate when treated with H₂O₂ may be increased by the addition of a compound containing a sulfhydryl group, i.e. cystine. A small part of this increase in rate of CO₂ is due to the action of the amino group as shown by the action of glycocoll. The results tend to show that the mode of action of H₂O₂ is one of dehydrogenation and that the action of the cystine is comparable to the Atmungskörper of Meyerhof.     

COMPARATIVE STUDIES ON RESPIRATION : XXIV. THE EFFECTS OF CHLOROFORM ON THE RESPIRATION OF DEAD AND OF LIVING TISSUE.

1. Chloroform in low concentration (0.25 per cent) causes an increase in the rate of production of CO₂ in Ulva; this is followed by a decrease. In higher concentration (0.5 per cent) only a decrease is observed. 2. Assuming that the normal oxidation depends on the action of peroxide and peroxidase, experiments were made by placing Ulva in 1.0 per cent H₂O₂ and in Fe₂(SO₄)₃ (which acts like a peroxidase). The former diminishes the rate, the latter increases and subsequently decreases it. 3. When Ulva is killed in such a manner as to destroy the oxidizing enzymes, no CO₂ is produced unless H₂O₂ and Fe₂(SO₄)₃ are present. If to this mixture chloroform is added, the effect depends on the concentration of the iron. If the concentration is low there is an increase in the production of CO₂ followed by a decrease. If the concentration is high the rate appears to decrease from the start.     

THE EFFECT OF LIGHT ON THE PERMEABILITY OF PARAMECIUM

1. The permeability of Paramecium to NH₄OH is greater when the cells are exposed to light than it is when they are in darkness. 2. This change can be demonstrated in cells exposed to monochromatic red light, though it is small. It becomes greater as the wave lengths shorten, and is greatest in the near ultra-violet. 3. The permeability increases as the duration of exposure to light is prolonged. 4. These experiments demonstrate the necessity of controlling the illumination when using Paramecium in physiological tests.     

SOME CHEMICAL ASPECTS OF THE POTASSIUM EFFECT

The ability of Nitella to distinguish electrically between Na⁺ and K⁺ (potassium effect) appears to depend on several organic substances (or groups of substances). Of these R_MK and R_SK determine the mobility and partition coefficient (S) respectively of K⁺ while R_MNa and R_SNa do the same for Na⁺. These substances can vary independently and this variation is susceptible to experimental control.     

METABOLISM AS RELATED TO CHROMOSOME STRUCTURE AND THE DURATION OF LIFE

This paper presents the rates of CO₂ production for four groups of Drosophila which differ in their chromosome constitutions. The four groups have metabolic rates which correlate with the balance of their chromosomes, the balanced chromosome groups of flies producing less CO₂ than the unbalanced chromosome groups. It is concluded therefore that genic balance plays a prominent part in metabolic control. The carbon dioxide rates are related to the duration of life within these groups. The results show that qualitatively the larger the production of CO₂ per day the shorter the time which the flies are capable of living. The agreement is not exact quantitatively. Rubner''s theory postulating a limit for the energy an organism is capable of metabolizing does not hold for the six classes of flies. The data show that the theory can be at most not more than a partial truth. Cell size is found to show no direct correlation with the metabolic rates of the different fly cohorts.     

THE IONIC ACTIVITY OF GELATIN

2.5 and 1.25 per cent gelatin have been titrated potentiometrically in the absence of salts and in the presence of two concentrations (0.0750 and 0.0375µ) of NaCl, MgCl₂, K₂SO₄, and MgSO₄. The data have been used to calculate values of ± S = v^z – (v – 1)^z, where v^z = v ² – (v ² – v) r_x/18. The maximum and minimum values of S with NaCl were used to calculate the mean distance (r_x) between like charges in gelatin. This is found to be 18 Å.u. or over (between acid or basic groups) which agrees with the probable value and the titration index dispersion. Thus the data with NaCl are shown to be normal and to obey the equation found to hold for simple weak electrolytes; namely, pK'' – pK = Sa See PDF for Equation where S is related to the valence and distance by the above equations. Using the NaCl data as a standard the deviations (ΔS) produced by the other salts are calculated and are found to agree quantitatively with the deviations calculated from equations derived for the simple weak electrolytes. This shows that in gelatin, as in the simple electrolytes, the deviations are related to the "apparent valences" (values which are a function of the true valence and the distance between the groups). The maximum "apparent valences" of gelatin are 2.4 for acid groups (in alkaline solution) and 1.8 for basic groups (in acid solution). These values correspond to the hypothetical condition of zero distance between the groups. They have no physical significance but have a practical utility first as mentioned above, and second in that they may be used in the unmodified Debye-Hückel equation to give the maximum effect of gelatin on the ionic strength. The true effect is probably even lower than these values would indicate. The data indicate that gelatin is a weak polyvalent ampholyte having distant groups and that the molecule has an arborescent structure with interstices permeated by molecules of the solvent and other solutes. The size and shape probably vary with the pH.     

THE KINETICS OF PENETRATION : V. THE KINETICS OF A MODEL AS RELATED TO THE STEADY STATE

An organic potassium salt, KG, passes from an aqueous phase, A, through a non-aqueous layer, B, into a watery solution, C. In C it reacts with CO₂ to form KHCO₃. The ionic activity product (K) (G) in C is thus kept at such a low level that KG continues to diffuse into C after the concentration of potassium becomes greater in C than in A. Hence potassium accumulates in C, the osmotic pressure rises, and water goes in. A steady state is eventually reached in which potassium and water enter C in a constant ratio. The rate of entrance of potassium (with no water penetrating into C) may fall off in a manner approximately exponential. But water enters and may produce an exponential decrease in concentration. This suggests that the kinetics may be treated like that of two consecutive monomolecular reactions. Calculations made on this basis agree very well with the observed values. The rate of penetration appears to be proportional to the concentration gradient of KG in the non-aqueous layer and in consequence depends upon the partition coefficients which determine this gradient. Exchange of ions (passing as such through the non-aqueous layer) does not seem to play an important rôle in the entrance of potassium. The kinetics of the model may be similar to that of living cells.