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1. In vitro preparations of rat jejunum were used to study the metabolic dependence, the structural specificity, and the pH sensitivity of the folic acid absorption process. 2. The presence of 2:4 dinitrophenol, and the absence of oxygen, in the muscosal bathing medium both led to a decrease in the tissue accumulation and serosal transfer of folic acid by everted sacs. 3. 10-formylfolic acid, present in the mucosal medium at a molar ratio of 10:1 with labelled folic acid, appears to compete for tissue accumulation sites but has no significant effect on serosal transfer. 4. The efflux of folic acid from pre-loaded jejunal sacs is stimulated by the presence in the mucosal medium of the folic acid and related compounds. Pteroyl-L-glutamic acid, 10-formylfolic acid and methotrexate elicit a significant increase in the efflux rate; pteroyl-D-glutamic acid was significantly less effective and pteroic acid had no effect. 5. The uptake of folic acid by isolated jejunal cells prepared by enzymic disaggregation of the mucosa, was found to be influenced by the pH of the bathing medium, uptake being enhanced at a pH of between 5 and 6. 6. It is concluded that the effect of metabolic inhibitors and acid pH conditions on the uptake of folic acid in vitro is consistent with a passive absorption mechanism, influenced in intact preparations and in vivo by the jejunal pH microclimate. However the occurrence of competitive inhibition and stimulated efflux may indicate the existence of a structurally specific accumulation process at some site, or sites, within the mucosa.

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