Endogenous proteins in the ileal digesta of adult humans given casein-, enzyme-hydrolyzed casein- or crystalline amino-acid-based diets in an acute feeding study.

BACKGROUND/OBJECTIVES: To ascertain if the form of dietary nitrogen (free amino acids (AA), small peptides, or intact protein) affects the endogenous nitrogen containing substances lost from the upper digestive tract of humans. SUBJECTS/METHODS: Digesta were collected via a naso-ileal tube from the terminal ileum of 16 adult humans in a single parallel study following an acute feeding regimen. Subjects were given an iso-nitrogenous and isocaloric test meal containing 150 g of casein (CAS) (n=6), enzyme-hydrolyzed casein (HCAS) (n=5) or crystalline AA (n=5) dissolved in 550 ml of water, as the sole sources of nitrogen. RESULTS: The mean concentrations and flows of total nitrogen, protein nitrogen, and soluble protein nitrogen passing the terminal ileum were significantly higher (P <0.01) for the CAS and HCAS test-meal groups compared to the AA meal group. Dietary CAS and HCAS had a considerable influence on digesta mucin concentrations and flows compared to free AA (+41%). Only 3-4% of the total nitrogen remained unidentified. CONCLUSIONS: The form of dietary nitrogen (protein, small peptides or free AA) had an acute effect upon the secretion or reabsorption of endogenous proteins in the small intestine of healthy humans, as evident from significant differences in both the quantity and composition of the proteins found in digesta at the end of the ileum.

Comparison of three markers for the determination of bacterial protein in terminal ileal digesta in the growing pig.

The aim of the study was to compare three methods commonly used to determine the concentrations of bacterial protein in digesta collected from the terminal ileum of growing pigs that had been fed a casein-based diet. The amounts of bacterial protein in terminal ileal digesta were determined using three different markers: 2.6-diaminopimelic acid (DAPA) and the d-amino acids, d-aspartic acid (d-Asp) and d-glutamic acid (d-Glu). The effectiveness of each marker was compared against a control based on physical fractionation by centrifugation. The total bacterial protein concentrations derived from the markers d-Asp and d-Glu were significantly different (p = 0.05) to those calculated from DAPA and the control, but there was no difference between DAPA and the control. The percentage of bacterial nitrogen ranged from 40% to 52% dependent on the marker used. Bacterial protein expressed as a percentage of the total protein, ranged from 48% to 62%, a substantial proportion of which (12-28%) was derived from lysed bacterial cells. Statistical correlations between the estimation methods were low. Such poor correlation between the markers may be the result of random errors such as variance in the epimerization of the two d-amino acids during protein hydrolysis. DAPA was accepted as a reliable marker for determining microbial protein in ileal digesta.

Surfactant inhibition of cholesterol oxidase.

Little is known of the effect of surfactants upon the activity of cholesterol oxidase. This study demonstrates the interrelationship of surfactant, enzyme and substrate, and illustrates a possible source of inaccuracy within an enzymatic cholesterol assay. Using the rate at which cholesterol is converted to delta 4-cholestenone, the reaction was followed directly bby monitoring the increase in absorbance at 240 nm. Inhibition of cholesterol oxidase was demonstrated with three surfactants, hydroxypolyethoxydodecane, Tween 20 and Triton-X-405. A fourth, Triton-X-100, produced high enzyme activity, although low concentrations resulted in incomplete substrate dispersal and high concentrations caused high blank values. Hydroxypolyethoxyduodecane was studied more closely and the mechanism of inhibition is suggested as poor substrate dispersal at low surfactant concentration and a competitive inhibition at higher concentrations.