STUDIES ON BLOOD COAGULATION : I. THE R?LE OF PROTHROMBIN AND OF PLATELETS IN THE FORMATION OF THROMBIN

1. A method is described for the preparation and titration of prothrombin and thrombin. 2. Confirming the views of Morawitz, Howell (1916–17, 1925), and Bordet, thrombin cannot be regarded as an artificial by-product of coagulation (Wooldridge, Nolf (both quoted from Morawitz)). Calcium, a platelet factor, and a plasma factor (prothrombin) interact to form thrombin, and this then acts upon fibrinogen to form fibrin. The amount and rate of thrombin formation in the first reaction are independent of the presence or absence of fibrinogen. After a variable latent period, thrombin suddenly appears in large quantities, coincident with or immediately preceding the deposition of fibrin if fibrinogen is present. 3. The amount of thrombin formed in a mixture of prothrombin, Ca and platelets is independent of the platelet or Ca concentration, and depends primarily upon the amount of prothrombin used. The platelets (or cephalin) enormously accelerate the transformation of prothrombin to thrombin, and this acceleration seems to be their physiological rôle in the coagulation process. 4. Contrary to previous reports, platelets have not been demonstrated to contain significant quantities of prothrombin. 5. The available data do not allow any definite decision as to whether the platelet factor actually combines with prothrombin to form thrombin, or merely catalyzes the transformation. The very slow formation of thrombin in the complete absence of platelets may be due to dissolved traces of platelet material released during the physical manipulation of the plasma (centrifuging, Berkefeld filtration). 6. There was no evidence for a species-specific activity of platelets in the transformation of prothrombin to thrombin.     

NOTE ON THE EXCITATION AND INHIBITION OF LUMINESCENCE IN BER?

1. The ions of Ca and K condition general luminescence, and are therefore necessary to the conduction of the impulse. 2. In van''t Hoff''s solution from which Mg is omitted, Berœ shows hyperirritability with respect to luminescence. This is the result of the action of Ca and K ions unantagonized by Mg. 3. The luminescent material spread on filter paper does not show luminescence in sea water, NaCl, MgCl₂, or saccharose solutions isotonic with sea water. In solutions of CaCl₂, SrCl₂, BaCl₂, KCl, and K₂SO₄ the indicator paper glows with a bright luminescence. 4. In dark adapted Berœ, luminescence is inhibited by a certain quantity of light. This quantity has an average value of 57,285 meter-candle-minutes, which is twelve times the value given by Mnemiopsis.     

WAVE LENGTH OF LIGHT AND PHOTIC INHIBITION OF STEREOTROPISM IN TENEBRIO LARV?

A definite intensity of white light is required (about 136 m.c.) to produce negative phototropic orientation of creeping Tenebrio larvæ away from contact with a vertical glass surface. This gives a measure of stereotropism in terms of phototropism, or reciprocally. The effectiveness of light for the suppression of stereotropism varies with wave length. It is therefore simple to obtain a measure of the relation between wave length and stimulating efficiency in this case of phototropic orientation. By determinations of the minimal energy required to inhibit stereotropism with different regions of the spectrum, it is found that the maximum effectiveness is sharply localized in the neighborhood of 535µµ. The curve connecting stimulating efficiency with wave length, while giving a picture of the effective absorption by the photosensory receptors, probably does not permit accurate characterization of the essential photosensitive material.     

THE R?LE OF PHOSPHATE IN BIOLOGICAL OXIDATIONS

1. The effect of phosphate on the oxidation of glyceric aldehyde by methylene blue, 1-naphthol 2-sulfonate indophenol, and phenol-indophenol has been studied. 2. At pH 4.77 in a phthalate-buffered medium phosphate does not catalyze the reaction. 3. At pH 7.9 in solutions buffered with borate, carbonate, or phenylalanine marked catalysis by phosphate is observed. The effect is most pronounced in borate. 4. Phosphate catalysis, within the limits studied, is strictly a linear function of the phosphate concentration. 5. The high concentration of HPO₄ ⁼ and the low concentration of PO₄ ^≡ relative to that of the substrate virtually demand the conclusion that the PO₄ ^≡ ion is the active catalytic species.     

THE R?LE OF THE ACTIVITY COEFFICIENT OF THE HYDROGEN ION IN THE HYDROLYSIS OF GELATIN

1. The hydrolysis of gelatin at a constant hydrogen ion concentration follows the course of a monomolecular reaction for about one-third of the reaction. 2. If the hydrogen ion concentration is not kept constant the amount of hydrolysis in certain ranges of acidity is proportional to the square root of the time (Schütz''s rule). 3. The velocity of hydrolysis in strongly acid solution (pH less than 2.0) is directly proportional to the hydrogen ion concentration as determined by the hydrogen electrode i.e., the \"activity;\" it is not proportional to the hydrogen ion concentration as determined by the conductivity ratio. 4. The addition of neutral salts increases the velocity of hydrolysis and the hydrogen ion concentration (as determined by the hydrogen electrode) to approximately the same extent. 5. The velocity in strongly alkaline solutions (pH greater than 10) is directly proportional to the hydroxyl ion concentration. 6. Between pH 2.0 and pH 10.0 the rate of hydrolysis is approximately constant and very much greater than would be calculated from the hydrogen and hydroxyl ion concentration. This may be roughly accounted for by the assumption that the uncombined gelatin hydrolyzes much more rapidly than the gelatin salt.     

THE R?LE OF ORGANIC SUBSTRATES IN PHOTOSYNTHESIS OF PURPLE BACTERIA

A representative of the photosynthetic non-sulfur purple bacteria (Athiorhodaceae) capable of using simple alcohols has been isolated in pure culture. By means of quantitative analysis of cultures at different stages of development it has been shown that this organism converts isopropanol quantitatively into acetone, simultaneously reducing CO₂ in the light. The data can be represented by the equation 2 CH₃CHOHCH₃ + CO₂ → 2 CH₃COCH₃ + (CH₂O) + H₂O. Manometric experiments with suspensions of resting cells have fully corroborated the results obtained with growing cultures. The experiments have conclusively proved that an organic substrate may fulfill exclusively the function of hydrogen donor for the photochemical CO₂-reduction in purple bacteria photosynthesis.     

MICRURGICAL STUDIES IN CELL PHYSIOLOGY : V. THE ANTAGONISM OF CATIONS IN THEIR ACTIONS ON THE PROTOPLASM OF AM?BA DUBIA.

I. The Plasmalemma. 1. On the plasmalemma of amebæ CaCl₂ antagonizes the toxic action of LiCl better than it does NaCl, and still better than it does KCl. MgCl₂ antagonizes the toxic action of NaCl better than it does LiCl and still better than it does KCl. 2. CaCl₂ antagonizes the toxic action of LiCl and of KCl better than does MgCl₂: MgCl₂ antagonizes NaCl better than does CaCl₂. II. The Internal Protoplasm. 3. The antagonizing efficiency of CaCl₂ and of MgCl₂ are highest against the toxic action of KCl on the internal protoplasm, less against that of NaCl, and least against that of LiCl. 4. CaCl₂ antagonizes the toxic action of LiCl better than does MgCl₂: MgCl₂ antagonizes the toxic action of NaCl and of KCl better than does CaCl₂. 5. LiCl antagonizes the toxic action of MgCl₂ on the internal protoplasm more effectively than do NaCl or KCl, which have an equal antagonizing effect on the MgCl₂ action. III. The Nature of Antagonism. 6. When the concentration of an antagonizing salt is increased to a toxic value, it acts synergistically with a toxic salt. 7. No case was found in which a potentially antagonistic salt abolishes the toxic action of a salt unless it is present at the site (surface or interior) of toxic action. 8. Antagonistic actions of the salts used in these experiments are of differing effectiveness on the internal protoplasm and on the surface membrane.