Food-grade expression of nattokinase in Lactobacillus delbrueckii subsp. bulgaricus and its thrombolytic activity in vitro.

OBJECTIVES: To produce nattokinase in a food-grade expression system and evaluate its thrombolytic activity in vitro. RESULTS: No nattokinase activity from reconstituted strains was observed in simulated gastric juice, but the enzyme was stable in intestinal fluid, the relative activity of which was found to be 60% after 4 h. Due to the nattokinase being produced intracellularly by recombinant bacterial strains, the persistence of the bacteria in gastric juice ensured transmission of the nattokinase into intestinal juice. Because of subsequent disintegration of the bacteria, the highest nattokinase activity was observed after 3 h at approximately 32%, following its carriage within the recombinant strains to the intestinal fluid. CONCLUSIONS: This study demonstrated that nattokinase from recombinant strains exhibited good thrombolytic activity in vitro and may be used by the dairy fermentation industry for the development of novel thrombolytic functional foods.

Mimicking the passage of avian influenza viruses through the gastrointestinal tract of chickens.

In contrast to human influenza viruses that replicate in the respiratory tract and are airborne transmitted, avian viruses also replicate in gut epithelial cells and are transmitted via the fecal-oral route. On this route, the virus is exposed to destructive fluids of the digestive tract, which are acidic and contain the proteases pepsin (gizzard) or chymotrypsin and trypsin (intestine). Only the latter enzyme activates virus by cleaving hemagglutinin (HA) into HA1 and HA2 subunits. We mimicked the passage of viruses through the gastrointestinal tract by treating them with digestive fluids from chicken and determined titers and integrity of HA by western-blot. Gizzard fluid completely inactivated virions and degrades HA even at a high dilution, but only if the pH was kept acidic. If the fluid is diluted with neutral buffer (mimicking virus uptake with seawater) particles were more resistant. Virions containing an uncleaved HA were even activated suggesting that gastric juice contains a trypsin-like protease. Undiluted intestinal fluid inactivated particles and destroyed HA, but diluted fluid activated virions. A virus isolated from the duck´s intestine is more tolerant against intestinal fluid compared to fowl plague virus suggesting that the former is better adapted to grow in the intestine. We also demonstrate that influenza viruses replicate to high titers in a novel chicken epithelial gut cell line. While viruses with a monobasic HA cleavage site require addition of trypsin, these cells effectively process HA with a polybasic cleavage site, which could be blocked with an inhibitor of the cellular furin protease.

Enhancement of Dissolution Rate and Intestinal Stability of Clopidogrel Hydrogen Sulfate.

BACKGROUND AND OBJECTIVES: Clopidogrel is an antiplatelet and antithrombotic prodrug. It has poor oral bioavailability due to poor dissolution and possible premature degradation in the intestine. Accordingly, the objective of this study was to enhance clopidogrel dissolution rate and to reduce its premature degradation in rabbit intestine. METHODS: Solid dispersion (SD) systems of clopidogrel with gelucire 50/13 and/or cremophor RH40 were prepared using fusion technique. The SD systems were characterized with respect to drug dissolution. The characterization included thermal analysis and infrared investigations. The stability of clopidogrel in the fluid extracted from small intestinal and colonic mucosal surfaces was monitored both in absence and presence of cremophor or gelucire. RESULTS: SD formation enhanced drug dissolution with the enhancement increasing at higher concentrations of either cremophor or gelucire. The ternary SD of clopidogrel with cremophor and gelucire reflected synergism between them. This synergism was manifested by enhanced dissolution efficiency of drug to reach 85 % at pH 6.8 and 89 % at pH 7.4 compared to unprocessed drug which liberated 16.2 and 15.2 % at the same pH values, respectively. Enhanced dissolution from SD was mainly due to micellar solubilization for cremophor and was due to change in the crystalline nature of drug with a contribution to self-emulsification in case of gelucire. Clopidogrel showed premature degradation in the intestinal fluid. Cremophor RH 40 reduced this degradation but gelucire failed in this respect. CONCLUSION: The study introduced SD system for enhanced dissolution rate of clopidogrel with a potential of reduced premature degradation in the intestine.

Investigation of the reactions of acrylamide during in vitro multistep enzymatic digestion of thermally processed foods.

This study investigated the fate of acrylamide in thermally processed foods after ingestion. An in vitro multistep enzymatic digestion system simulating gastric, duodenal and colon phases was used to understand the fate of acrylamide in bakery and fried potato products. Acrylamide levels gradually decreased through gastric, duodenal and colon phases during in vitro digestion of biscuits. At the end of digestion, acrylamide reduction was between 49.2% and 73.4% in biscuits. Binary model systems composed of acrylamide and amino acids were used to understand the mechanism of acrylamide reduction. High-resolution mass spectrometry analyses confirmed Michael addition of amino acids to acrylamide during digestion. In contrast to bakery products, acrylamide levels increased significantly during gastric digestion of fried potatoes. The Schiff base formed between reducing sugars and asparagine disappeared rapidly, whereas the acrylamide level increased during the gastric phase. This suggests that intermediates like the Schiff base that accumulate in potatoes during frying are potential precursors of acrylamide under gastric conditions.

Impact of extraneous proteins on the gastrointestinal fate of sunflower seed (Helianthus annuus) oil bodies: a simulated gastrointestinal tract study.

In this study, we examined the physicochemical nature of sunflower seed oil bodies (in the absence and presence of added protein) exposed to gastrointestinal conditions in vitro: crude oil bodies (COB); washed oil bodies (WOB); whey protein isolate-enriched oil bodies (WOB-WPI); and, sodium caseinate enriched-oil bodies (WOB-SC). All oil body emulsions were passed through an in vitro digestion model that mimicked the stomach and duodenal environments, and their physicochemical properties were measured before, during, and after digestion. Oil bodies had a positive charge under gastric conditions because the pH was below the isoelectric point of the adsorbed protein layer, but they had a negative charge under duodenal conditions which was attributed to changes in interfacial composition resulting from adsorption of bile salts. Oil bodies were highly susceptible to flocculation and coalescence in both gastric and duodenal conditions. SDS-PAGE analysis indicated degradation of oleosin proteins (ca. 18-21 kDa) to a greater or lesser extent (dependent on the emulsion) during the gastric phase in all emulsions tested; there is evidence that some oleosin remained intact in the crude oil body preparation during this phase of the digestion process. Measurements of protein displacement from the surface of COBs during direct exposure to bile salts, without inclusion of a gastric phase, indicated the removal of intact oleosin from native oil bodies.

Silica nanoparticle stabilization of liquid crystalline lipid dispersions: impact on enzymatic digestion and drug solubilization.

The high internal surface area and drug solubilizing capacity of liquid crystal lipids makes them promising oral drug delivery systems. Pluronic F127 is typically used to disperse highly viscous cubic liquid crystal lipids into cubosomes; however, such copolymers alter the internal structure and provide little control over enzymatic digestion. This study aimed to use hydrophilic silica nanoparticles to stabilize glyceryl monooleate (GMO) cubosomes prepared by ultrasonication. We investigate the influence of silica nanoparticles size and concentration on the physical (colloidal) and chemical (enzymatic digestion) stability, as well as in vitro solubilization of cinnarizine as a poorly soluble model drug. Silica stabilized nanostructured liquid crystal dispersions (120 nm to150 nm in diameter and zeta potentials of-30 mV to -60 mV) were successfully prepared with excellent long-term stability (<10% size change after 30 days). Silica stabilized GMO cubosomes demonstrated reduced enzymatic digestion compared to pluronic F127 stabilized cubosomes. This reduced digestion was attributed to a combination of adsorbed silica nanoparticles acting as a physical barrier and excess dispersed silica adsorbing/scavenging the lipase enzyme. Under simulated intestinal digestion conditions, silica stabilized GMO cubosomes showed a greater solubilization capacity for cinnarizine, which precipitated in non-crystalline form, in comparison to pure drug suspensions or pluronic F127 stabilized GMO cubosomes. Silica nanoparticle stabilized GMO liquid crystal dispersions are a promising oral delivery vehicle.

Ex vivo digestion of carp muscle tissue--ACE inhibitory and antioxidant activities of the obtained hydrolysates.

In the digestive tract of humans, bioactive peptides, i.e. protein fragments impacting the physiological activity of the body, may be released during the digestion of food proteins, including those of fish. The aim of the study was to establish the method of human ex vivo digestion of carp muscle tissue and evaluate the angiotensin I-converting enzyme inhibitory and antioxidant activities of hydrolysates obtained after digestion. It was found that the hydrolysates of carp muscle tissue obtained with the three-stage method of simulated ex vivo digestion showed ACE inhibitory as well as antioxidative activities. It was demonstrated that the degree of hydrolysis depended on the duration of individual stages and the degree of comminution of the examined material. Although the applied gastric juices initiated the process of hydrolysis of carp muscle tissue, the duodenal juices caused a rapid increase in the amount of hydrolysed polypeptide bonds. The antihypertensive and antioxidative activities of the hydrolysates of carp muscle tissue increased together with progressive protein degradation. However, the high degree of protein hydrolysis does not favour an increase in the activity of free radical scavenging. The presented results are an example of the first preliminary screening of the potential health-promoting biological activity of carp muscle tissue in an ex vivo study.

Impact of solidification on the performance of lipid-based colloidal carriers: oil-based versus self-emulsifying systems.

The study aims to develop and optimise lipid-based colloidal carriers (LBCC) for enhancing solubilisation and reducing fed/fasted variation for the poorly water-soluble danazol (DAN). Oil-based and self-microemulsifying delivery systems (SMEDDS) were developed, and the effect of solidification was investigated. Liquid SMEDDS (L-SMEDDS, Capmul MCM:Tween 80:Transcutol HP 1:2:1, w/w) and emulsion (Capmul MCM:soya lecithin 100:0.6, w/w) were developed. Solid-state formulations were prepared via (i) physical adsorption of L-SMEDDS (P-SMEDDS) or (ii) spray drying of emulsion (silica-lipid hybrid, SLH) and L-SMEDDS (spray-dried SMEDDS, S-SMEDDS) using Aerosil 380 silica nanoparticles as the solid carrier. In vitro lipid digestion and drug solubilisation under simulated intestinal conditions in both fasted and fed states were investigated. Solubilisation of unformulated DAN under both fasted and fed conditions was low, and a large fed/fasted variation was observed, i.e. 6.6-fold difference. All LBCC formulations provided enhanced drug solubilisation and significantly reduced the fed/fasted variation. For self-emulsifying LBCC, the fasted state drug solubilisation was ranked as L-SMEDDS > PSMEDDS > S-SMEDDS, suggesting that solidification reduced the capability of SMEDDS in presenting DAN to the aqueous phase. However, in the case of oil-based LBCC, improved drug solubility was observed with the solid form SLH under both fasted and fed state in comparison to that of the equivalent liquid form. Overall, the SLH, which provided the highest drug solubilisation in the fasted state (i.e. 10-fold higher than the pure DAN) and the smallest fed/fasted variation, was considered an optimised solid LBCC to enhance the solubilisation of DAN and reduce the fed/fasted variation.

Enhancing vitamin E bioaccessibility: factors impacting solubilization and hydrolysis of α-tocopherol acetate encapsulated in emulsion-based delivery systems.

Oil-soluble vitamins are often encapsulated within emulsion-based delivery systems to facilitate their incorporation into aqueous-based products. We have examined the influence of carrier oil type and simulated small intestinal fluid (SSIF) composition on the bioaccessibility of emulsified vitamin E using a gastrointestinal model. Oil-in-water emulsions containing vitamin E acetate were prepared using bile salts as emulsifier, and either long chain triacylglycerols (glyceryl trioleate, LCT) or medium chain triacylglycerols (glyceryl trioctanoate, MCT) as carrier oils. The addition of calcium (CaCl2) to the SSIF increased the extent of lipid digestion in LCT-emulsions, but had little impact in MCT-emulsions. The bioaccessibility of vitamin E increased in the presence of calcium and phospholipids (DOPC) in LCT-emulsions, but decreased in MCT-emulsions. The highest bioaccessibility (≈66%) was achieved for LCT-emulsions when the SSIF contained both calcium and phospholipids. The conversion of α-tocopherol acetate to α-tocopherol after in vitro digestion was considerably higher for LCT-emulsions when calcium ions were present in the SSIF, but was not strongly affected by SSIF composition for MCT-emulsions. In general, this research provides important information about the factors influencing the bioaccessibility of emulsified vitamin E, which could be used to design more effective emulsion-based delivery systems for increasing the oral bioavailability of this important bioactive component.

In vitro gastric and intestinal digestion of a walnut oil body dispersion.

An oil body dispersion (11.3% fat) was prepared by wet disintegration of walnuts and was then subjected to a two-step model of in vitro digestion. In a gastric environment, proteolysis by pepsin led to the destabilization and coalescence of the oil bodies. Aggregation of the coalesced oil bodies was apparent under a confocal microscope, with aggregates up to 275 µm in size. Pepsin-resistant peptides and proteins remained at the surface of the oil bodies, and some were further resistant to intestinal proteases. Under intestinal conditions, the hydrolysis of walnut triglycerides led to the spontaneous formation of a new type of multiple emulsions, ranging from 2 to 45 µm in size and with protein material inside the inner water droplets. Transmission electron microscopy revealed the presence of a liquid-crystalline phase of bile salts and lipolytic products at the surface of the oil droplets and some bile salt crystals at the surface of the inner water droplets.

Assessment of the sequential simulated gastrointestinal tolerance of lactic acid bacteria from kefir grains by response surface methodology.

The tolerance of lactic acid bacteria (LAB) from kefir grains to gastrointestinal tract conditions was evaluated in vitro. The effects of pH values and bile salts on the viability of LAB were investigated. The results demonstrated that pH value showed a significant effect on the viability. The viable counts exhibited a reduction of 1.5 to 2 log cycles in 0.3% to 0.5% bile salts after 4 h. The viability of LAB exposure to sequential simulated gastric and intestinal juices was assessed by response surface model (RSM). RSM indicated that the gastric pH and gastric contact time significantly affected the viability (P < 0.05), while the effect of intestinal contact time was not significant. Moreover, RSM revealed the interactions of pH and gastric contact time, and of pH and intestinal contact time. The LAB cells, temporarily damaged by the low pH of gastric juice (pH < 2), could recover in the intestinal juice; and the longer the intestinal contact time, the higher the viability of LAB. RSM proved to be a useful and accurate method to predict the viability of LAB under certain laboratory conditions by the model validation. This study indicated that LAB from kefir grains exhibited excellent tolerance to sequential simulated gastrointestinal tract conditions, and that kefiran possessed a significant protective effect on LAB in hostile environments.

Characterization of insulin protection properties of complexation hydrogels in gastric and intestinal enzyme fluids.

The objective of this study was to elucidate the mechanisms contributing to oral bioavailability of insulin by poly(methacrylic acid grafted with poly(ethylene glycol)) (P(MAA-g-EG)) hydrogels using the gastric and intestinal fluids from rats. P(MAA-g-EG) hydrogels successfully protected the incorporated insulin from enzymatic degradation by forming interpolymer complexes in the gastric fluid. The hydrogels also showed the insulin protection ability by itself. In the intestinal fluid, P(MAA-g-EG) hydrogels significantly decreased the insulin degradation rate and calcium ion levels, while protein levels was not changed. Insulin protecting effects were dependent on the fraction of the carboxylic group in the polymer networks. Moreover, the insulin degradation inhibitory effect was significantly correlated with Ca2+ deprivation ability of P(MAA-g-EG) hydrogels in the intestinal fluid, implying that the Ca2+ deprivation ability plays an important role in the inhibition of the intestinal enzyme activities. Insulin-loaded P(MAA-g-EG) (ILPs) hydrogels showed a rapid and almost complete insulin release even in the presence of intestinal proteases. These results suggested that the insulin protection ability of the hydrogels contributed to improve oral insulin absorption and that P(MAA-g-EG) hydrogels can be an excellent carrier for protecting insulin during their transit through the GI tract.

Digestive stimulant action of spices: a myth or reality?

Spices have long been recognized for their digestive stimulant action. Several spices are also employed in medicinal preparations against digestive disorders in traditional and Indian systems of medicine. Earlier reports on the digestive stimulant action of spices are largely empirical; only in recent years, this beneficial attribute of spices has been authenticated in exhaustive animal studies. Animal studies have shown that many spices induce higher secretion of bile acids which play a vital role in fat digestion and absorption. When consumed through the diet also spices produce significant stimulation of the activities of pancreatic lipase, amylase and proteases. A few of them also have been shown to have beneficial effect on the terminal digestive enzymes of small intestinal mucosa. Concomitant with such a stimulation of either bile secretion or activity of digestive enzymes by these spices, leading to an accelerated digestion, a reduction in the food transit time in the gastrointestinal tract has also been shown. Thus, the digestive stimulant action of spices seems to be mediated through two possible modes: (i) by stimulating the liver to secrete bile rich in bile acids, components that are vital for fat digestion and absorption, and (ii) by a stimulation of enzyme activities that are responsible for digestion. This review highlights the available information on the influence of spices on the digestive secretions and enzymes.

[The effects of an oral calcium overload on the rat exocrine pancreas]. / Efectos de la sobrecarga oral de calcio sobre el páncreas exocrino de rata.

AIM: The possible induction of functional or morphologic changes in the exocrine pancreas of the rat by oral calcium overload was studied to determine the possible relationship to predisposition of acute pancreatitis over the acinar theory. MATERIALS AND METHODS: Oral chloride calcium (0.45 and 0.25 g/kg body weight/day) plus cholelecalciferol (300,000 UI/kg i.m.) were administered to male Wistar rats over 1 to 3 months. Half of each group, including a control were submitted to cholinergic stimulation with carbamylcholin. After anesthesia, blood and pancreatic tissue and duodenal fluid were extracted for enzymatic and ultrastructural studies. RESULTS: In the rats treated with high doses of calcium for 1 month greater tissue concentrations of amylase, lipase and trypsin were observed. Moreover, there was a greater trend to the presence of dilated ergastoplasm. In the rats treated with high doses over 3 months a lower enzyme concentration was observed in the animals not stimulated that in the control group. On stimulation with carbamylcholin, higher concentrations of enzymes were observed in tissue than in those not stimulated. This was accompanied by a lower number of exocytosis in this experimental group that in the control. CONCLUSIONS: A possible increase in the calcium concentration in the acinar cell may lead to dysfunction in the secretory mechanisms favoring the intracellular accumulation of digestive enzymes, predisposing intracellular activation, in the context of the acinar or lysosomal hypothesis of acute pancreatitis.

[Detection of telomerase activity in intestinal lavage solution from colorectal carcinoma patients].

In terms of colorectal carcinoma, the fecal occult blood test is widely used for mass survey, but has many complicated problems to be overcome. Telomerase activity has been reported in a wide range of malignancies. We have examined telomerase activity of intestinal lavage solution collected from 16 colorectal carcinoma patients and from 10 volunteers (control) by the method of telomeric repeat amplification protocol (TRAP) assay. Patients drunk polyethylene glycol-electrolyte lavage solution (PEG-ELS) before examination. Sample solutions were collected by colonoscope at the beginning of colonoscopy. The telomerase activity from colorectal carcinoma patients were positive 9/16 (56.3%) including 2 cases of early stage. In volunteers, were positive 1/10 (10.0%). This method has, therefore, possibility for a new useful method of diagnosis for colorectal carcinoma.

pH and concentrations of pepsin and trypsin in feeding tube aspirates as predictors of tube placement.

BACKGROUND: The purpose of this study was to determine the extent to which pepsin and trypsin concentrations in feeding tube aspirates, in addition to pH, contribute to predicting feeding tube position. METHODS: Aspirates from 742 feeding tubes (nasogastric, n = 343; nasointestinal, n = 399) were tested for pH and enzyme concentrations. Also tested were aspirates from two feeding tubes inadvertently positioned into the lung (one in the pleural space and one in the tracheobronchial tree) and 146 samples of tracheobronchial and pleural fluids collected by other methods. Enzyme assays were conducted in a research laboratory. Results of the pH and enzyme tests were compared with radiologic reports of tube location. RESULTS: Gastric fluid had a mean low pH (4.06), a high mean pepsin concentration (349.1 micrograms/mL), and a low mean trypsin concentration (19.3 micrograms/mL). In contrast, intestinal fluid had a mean high pH (7.40), a high mean trypsin concentration (143.0 micrograms/mL), and a low mean pepsin concentration (24.2 micrograms/mL). Respiratory samples also had a high mean pH (7.89) but contained little or no pepsin or trypsin. Using a logistic regression equation with all three variables to differentiate between respiratory and gastrointestinal placement, it was possible to correctly classify 100% of the respiratory cases and 93.4% of the gastrointestinal cases. Another equation used to differentiate between gastric and intestinal sites was able to classify correctly 91.2% of the gastric cases and 91.5% of the intestinal cases. CONCLUSIONS: The results clearly indicate that laboratory-determined enzyme concentrations in feeding tube aspirates are helpful in predicting tube location. Thus, it is desirable that inexpensive, simple bedside tests be developed so that they can be used in conjunction with pH measurements to help predict tube position.

Method of measurement of pancreatic elastase II activity and postnatal development of proteases in human duodenal juice and bovine and porcine pancreatic tissue.

A specific method for pancreatic elastase II activity analysis was developed. True elastase II activity could be discriminated from that of elastase I and chymotrypsin. The postnatal development of four pancreatic proteases in the duodenal juice of children and in the pancreatic homogenates of calves and piglets was measured. The study was carried out on patients without (14 children) and with (5 children) pancreatic insufficiency. Calves and piglets were either milk-fed or weaned until slaughter at different ages. Profiles of enzyme development were globally similar in milk-fed piglets and calves, while in children without pancreatic insufficiency, no significant change was observed between 4 and 168 months. In children with pancreatic insufficiency, enzyme activity was low. In animals, elastase II and chymotrypsin activities were maximal at birth, decreased with age, and probably were associated with the digestion of milk protein. In contrast, elastase I and trypsin activities increased markedly after weaning in connection with the intake of solid food.

Physiological control of cholecystokinin release and pancreatic enzyme secretion by intraduodenal bile acids.

BACKGROUND: The physiological relevance of duodenal bile acids in the control of cholecystokinin release and pancreatic enzyme secretion is still unknown. AIMS: To provide a near physiological situation by perfusing a bile acid mixture mimicking the individual endogenous bile acid composition of the person under investigation. For maximal reduction of endogenous bile output the CCK-A receptor antagonist loxiglumide was infused intravenously. SUBJECTS AND METHODS: Seven healthy volunteers were studied on four different days by a duodenal marker perfusion technique. The individual bile acid composition in duodenal juice and test meal stimulated bile acid output was assessed on day 1. Bile acids were perfused at an amount of 30 or 100% as determined on day 1 in combination with the test meal in the presence or absence of loxiglumide. Pancreatic enzymes, bilirubin, and bile acid output were determined in duodenal juice. Plasma cholecystokinin (CCK) and plasma pancreatic polypeptide (PP) were measured radioimmunologically. RESULTS: Bile acid perfusion did not significantly alter stimulated pancreatic enzyme, bilirubin or bile acid output or plasma CCK. Loxiglumide did not alter basal CCK release but increased test meal stimulated CCK output fourfold (p < 0.05). The addition of bile acids to the test meal at a dose resembling 30% of bile acid output as determined on day 1 prevented this increase. Plasma PP concentration remained unchanged by bile acids and were mostly undetectable during loxiglumide infusion. CONCLUSIONS: The CCK producing cell is under constant suppression by intraduodenal bile acids which cannot be further enhanced by a physiological bile acid mixture. However, removal of duodenal bile acids by inhibition of gall bladder contraction unmasks this suppression leading to a dramatic increase in plasma CCK levels. As little as one third of postprandially released bile acids completely reverse this effect. Bile acids are the most important luminal regulator of CCK release in humans.

[Measurement of fecal elastase 1 by immunoreactivity: a new indirect test of the pancreatic function]. / Mesure de I'élastase 1 fécale par immunoréactivité: une nouvelle approche indirecte de la fonction pancréatique.

AIM: The aim of this study was to evaluate the potential and precision of the fecal elastase type 1 rest in comparison to the secretin-pancreozymin-test in the diagnosis of exocrine pancreatic insufficiency. METHODS: We studied 254 stool samples from 102 individuals without malabsorption (n = 53) and patients with pancreatic maldigestion syndromes (n = 49). Pancreatic elastase was measured immunologically, using a new enzyme immunoassay according to the sandwich technique. RESULTS: Spot stool immunoreactive elastase activity in controls ranged from 136 to 4,400 micrograms/g. Ninety five percent of all values where within 175 to 1,500 micrograms/g. The lower limit of normal was defined as 150 micrograms/g. No significant decrease of immunoreactivity was found when stool samples were stored at room temperature over five days. The assay variability calculated from 10 consecutive assays of a single fecal sample gave coefficients of variation ranging from 3.3 to 6.3% for intraassay-variability and from 4.1 to 10.2% for interassay-variability. There was a good correlation between the output of elastase compared to lipase output with correlations coefficients of 0.821 in controls and 0.905 in patients with impaired pancreatic function. In stool samples of 49 patients with exocrine pancreatic insufficiency the concentration of fecal elastase was significantly lower (P < 0.001) compared to controls and patients with Crohn or coeliac disease. Elastase immunoreactivity showed higher sensitivity and specificity as compared to fecal chymotrypsin. Furthermore, in contrast to fecal chymotrypsin, the test results were unaffected by pancreatic enzyme replacement therapy. CONCLUSION: These results indicate that fecal immunoreactive elastase may be recommended as a new, non-invasive easy-to-perform tubeless pancreatic function test with a high sensitivity and specificity in comparison with healthy controls.

Growth, viscosity and beta-glucanase activity of intestinal fluid in broiler chickens fed on barley-based diets with or without exogenous beta-glucanase.

1. Three groups of birds were fed for up to 35 days on diets containing 500 g barley (cv. Condor)/kg diet, with or without exogenous beta-glucanase, either a commercial preparation or a recombinant endoglucanase. 2. Birds which received diets containing the exogenous enzymes grew faster for the first 3 weeks but after that there was no apparent difference in rate of growth. 3. beta-Glucanase activities in the crop and small intestine of birds given exogenous enzymes were generally higher than those of birds given only the basal diet. 4. Viscosity of intestinal fluid in birds given only the basal diet decreased with age but there was no corresponding increase in beta-glucanase activity. This discounts bacterial beta-glucanase as a contributory factor in the adaptation to beta-glucanase apparent in older birds.