1. The acceleration by bicarbonates of the swelling and hemolysis of erythrocytes in solutions of ammonium salts, first reported by Ørskov, is strikingly dependent upon carbonic anhydrase, being almost abolished by inhibitors of this enzyme such as KCN and sulfanilamide, and under suitable conditions being enhanced by its addition to the external solution. This behavior gives support to the theory of "catalyzed diffusion" as an explanation of the Ørskov effect.
2. The inhibitory effects of both sulfanilamide and KCN seem to be capable of complete reversal on washing the erythrocytes in isotonic salt solutions. The full effect of KCN appears almost instantly; that of sulfanilamide requires a period measured in seconds, or possibly even in minutes, to reach its maximum, the delay presumably being due to the slower penetration of the erythrocyte by this substance. Under favorable conditions the effect of concentrations of sulfanilamide of a few hundredths of a milligram per cent can be demonstrated. No similar effects have been obtained with sulfapyridine.
3. Bicarbonates also have a "catalytic" effect on the response of the internal pH of erythrocytes to changes in that of their surroundings. The resulting volume changes of the cell, which otherwise frequently require many minutes for their completion, may take place within a few seconds in the presence of low concentrations of bicarbonates. At a given pH value the effect of the latter substances is chiefly on the rate of the change and only to a minor extent on its magnitude. It may be further accelerated under appropriate conditions by the addition to the cell suspension of carbonic anhydrase, and can be almost abolished by KCN and by sulfanilamide.
4. Volume changes of erythrocytes associated with exchanges of Cl' for SO4'' ions are greatly accelerated by low concentrations of bicarbonates, this effect being likewise dependent upon carbonic anhydrase. There is some evidence that in this case the exchange takes place, at least in part, in two steps: Cl' for HCO3' and HCO3' for SO4''.
