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Quantitation of major protein constituents of murine intestinal fluid
Authors:Steffen Bade  Sabine Koelling  Fabian Reuter  Andreas Frey
Affiliation:a Division of Mucosal Immunology, Research Center Borstel, 23845 Borstel, Germany
b Central Chemical Analytical Laboratory, Technical University, 21073 Hamburg, Germany
c Department of Pneumology, Research Center Borstel, 23845 Borstel, Germany
Abstract:The gastrointestinal tract is a hostile biological environment, yet not all ingested materials are destroyed. The minute differences that determine whether a substance persists or is digested, liberated, adsorbed, excreted, or taken up are still poorly understood. Most attempts to investigate the events occurring during an orogastrointestinal passage rely on simplified in vitro systems where an analyte is exposed to artificial intestinal fluids. To closely mimic the events in the gastrointestinal tract, the exact intestinal fluid composition and the in vivo concentration of its constituents must be known. The widely used lavage procedures, however, dilute the intestinal fluids to an extent that precludes recalculation to the original concentrations. Thus, we developed procedures with which undiluted murine intestinal fluid can be harvested; determined the in vivo concentrations of the digestive enzymes trypsin, chymotrypsin, and elastase and the adsorbents mucin and immunoglobulin A in small intestinal fluid of fasted and unfasted female Balb/c mice; and identified chymotrypsin and immunoglobulin A as valid endogenous dilution markers for the recalculation of aqueous lavages. With these technologies and information at hand, more reliable investigations on the fate of allergens, pathogens, food, and anthropogenic xenobiotics in the gastrointestinal tract will be possible.
Keywords:Gastrointestinal passage   Intestinal lavage   Immunoglobulin A (IgA)   Mucin   Trypsin   Chymotrypsin   Elastase   Perfluorohexane   In vivo concentration   Dilution marker   Digestion   Adsorption
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