Enterokinase initiates digestion of protein by conversion of trypsinogen into trypsin. The interactions between enterokinase and trysin were investigated in 6 patients with intractable diarrhea of infancy and 34 children with celiac disease. The six infants between 2 and 3 months with intractable diarrhea of infancy had reduced mucosal enterokinase activity (9.5 +/- 4.8muM per gram of protein per minute) and reduced intraluminal trypsin activity (2.9 +/- 0.7muM per gram of protein per minute) as compared with healthy controls (109 +/- 34.2muM per gram of protein per minute and 14.3 +/- 5.8muM per gram of protein per minute) respectively. The activities of all enzymes returned toward normal following treatment with intravenous alimentation. The mucosal morphology of all pretreatment biopsies in all cases showed Grade III atrophy which improved. These findings suggest that enterokinase deficiency and reduced intraluminal trypsin activity in intractable diarrhea of infancy may be one of the contributing factors to protein malabsorption and consequent malnutrition. Thirty-four children with celiac disease were between the age of 9 months and 13 years. The 11 newly diagnosed patients with celiac disease demonstrated Grade III to IV atrophy of the mucosa. The 23 patients with treated celiac disease on a gluten-free diet showed a normal to Grade II atrophy. In both treated and untreated celiac disease the enterokinase activities and the intraluminal trypsin activity were within normal limits. The enterokinase activity in celiac disease is near normal in contrast to the marked reduction noted in intractable diarrhea of infancy even though the intestinal mucosa shows marked morphological alteration and the disaccharidase activities are greatly reduced in celiac disease. After a prolonged alimentary fast of up to 26 days on intravenous alimentation, two patients with intractable diarrhea of infancy showed improvement in the activities of enterokinase and trypsin. These findings demonstrate that enterokinase and trypsin activities in the gut were present and improved in the absence of oral feeding.