The rate of hydrochloric acid production by isolated, bullfrog gastric mucosae depends critically on the supply of chloride ion to the serosal surface. Secretion of acid is negligible if chloride is completely replaced by glucuronate and gluconate ion. The experimental evidence indicates that the rate of acid secretion may be regarded as a reaction velocity, depending on chloride concentration in a manner closely resembling Michaelis-Menten kinetics. Bromide and iodide ions substitute, in varying degree, for chloride as substrate. A familiar inhibitor of gastric acid production, thiocyanate ion, appears to act by competition with chloride in a reaction leading to the formation of acid. This reaction is included in a hypothetical reaction cycle, generalized from the redox model for gastric acid production. Under certain conditions, the model predicts a dependence of secretion rate on chloride supply of the Michaelis-Menten type, as was observed.

Direct measurements have been made of the net volume flow through cellulose membranes, due to a difference in concentration of solute across the membrane. The aqueous solutions used included solutes ranging in size from deuterated water to bovine serum albumin. For the semipermeable membrane (impermeable to the solute) the volume flow produced by the osmotic gradient is equal to the flow produced by the hydrostatic pressure RT ΔC, as given by the van't Hoff relationship. In the case in which the membrane is permeable to the solute, the net volume flow is reduced, as predicted by the theory of Staverman, based on the thermodynamics of the steady state. A means of establishing the amount of this reduction is given, depending on the size of the solute molecule and the effective pore radius of the membrane. With the help of these results, a hypothetical biological membrane moving water by osmotic and hydrostatic pressure gradients is discussed.

The total active transport of chloride ions across the gastric mucosa can be considered as the sum of two fractions; an acidic one which is equivalent to the acid secreted, and an electromotive one which accounts for the electric energy generated by the gastric mucosa. In the present studies, the relationship between this electromotive chloride transport and acid secretion has been investigated, using specific inhibitors. The rate of electromotive chloride transport was found to be essentially unaffected by changes in the rate of acid secretion, and also by inhibition of acid secretion by thiocyanate. On the other hand, diamox, in combination with histamine, was shown to depress or abolish the gastric electromotive force and to inhibit partially the total chloride transport, while acid was secreted at an almost normal rate. This kind of inhibition is undefined as to its mechanism but seems to be more specific for the gastric chloride transport than any other inhibitor known. It is concluded that acid secretion and electromotive chloride transport involve two different mechanisms, and are not absolutely essential for each other. The present results do not support the view that carbonic anhydrase is essential for acid secretion. They rather suggest an important function of this enzyme in the mechanism of active chloride transport.

The unidirectional fluxes of Cl - and Na + across the frog gastric mucosa in vitro were investigated with radioactive isotopes, and related to the secretory and electrical properties of the normal, and metabolically inhibited, mucosa. The flux of Cl - from nutrient to secretory surface of the mucosa was observed to rise sharply with increasing acid secretion, while the corresponding flux of Na + did not change appreciably. Lowering [NaCl] in the secretory solution caused a proportional drop in the fluxes from secretory to nutrient surface, of both Cl - and Na + . Under the same conditions, the flux of Cl - from nutrient to secretory surface fell by nearly the same amount as did the flux of Cl - in the opposite direction, while the flux of Na + from nutrient to secretory surface remained essentially unchanged. Electrical and hydrodynamic causes for this observation could be excluded. Metabolic inhibitors, including cyanide, azide, DNP, and anaerobiosis depressed Cl - flux in both directions distinctly below the corresponding values observed with the normal, non-secreting mucosa. At the same time, a decrease in electrical potential difference and conductance was observed under inhibition. The flux of Na + was little changed by metabolic inhibition. The relationship between fluxes and conductance of Cl - during metabolic inhibition differs markedly from that observed under normal conditions, and is consistent with the view that during metabolic inhibition most of the Cl - moving across the mucosa does so as a free ion. From the above data it is concluded that Cl - is normally transported across the mucosa in combination with a carrier, the supply of which is impaired under metabolic inhibition. According to the behavior of the Na + flux, the passive permeability of the mucosa appeared to be little affected by the metabolic inhibition applied, but seemed to rise considerably after death of the mucosa, probably due to structural damage.

To elucidate the role of protein synthesis in DNA formation, E. coli R2 infected with phage T2 was studed as a model, employing chloramphenicol to inhibit protein synthesis. The following results were obtained. 1. Chloramphenicol inhibited protein synthesis but not synthesis of nucleic acids in uninfected bacteria. 2. Studies of the effect of chloramphenicol on phage maturation indicated a delay of 2 minutes between time of addition and cessation of phage growth. 3. The increase of DNA in phage-infected bacteria was completely suppressed by the addition of chloramphenicol within 2 minutes following infection. Addition at later times showed progressively less inhibitory action depending upon the time interval, and addition after the 10th or 12th minute showed no appreciable effect on DNA synthesis despite the cessation of intracellular phage formation and protein synthesis. 4. When chloramphenicol was added to infected cells the increase of resistance to UV stopped within 2 minutes, whether or not DNA synthesis continued. Thus evolution of resistance paralleled the rate of DNA synthesis achieved, but not the amount of DNA accumulated. 5. We conclude that in infected bacteria, protein synthesis is necessary to initiate DNA synthesis but is not essential for its continuation. The resistance to UV that characterizes infected cells near the midpoint of the latent period is not due to accumulation of DNA, but depends on some chloramphenicol-sensitive process (probably protein synthesis) completed at about the time the rate of DNA synthesis becomes maximal.