Stimulating effect of pancreatic-like enzymes on the development of the gastrointestinal tract in piglets.

Use of nutritional components from the milk and eventually from the solid feed relates to the growth and development of gastrointestinal tract (GIT). We studied the effect of pancreatic-like enzymes [porcine pancreatic enzymes (Creon) or microbial-derived amylase, protease, and lipase] on GIT morphology and lipid absorption in suckling piglets. Both enzyme preparations, in low or high dose, were fed via a stomach tube twice a day for 7 d starting at 8 d of age and controls received vehicle, n = 6. The day after treatments ended, lipid absorption was tested after which pigs were euthanized and GIT was examined. Enzyme cocktails, irrespective of their origin, increased (P < 0.001) triglyceride level in blood. Enzyme preparation affected (P < 0.001) small intestinal mucosal thickness, villi length, and crypt depth and (P < 0.01) mitotic division of enterocytes. In addition, the external administration of pancreatic enzymes stimulated pancreatic growth as observed by increased (P < 0.05) mitotic division of pancreatic cells. The study revealed that pancreatic or pancreatic-like enzymes of microbial origin administrated in the early postperinatal period enhance GIT development and may be used to better prepare the GIT of piglets for milk use and weaning.

Growth is dependent on the exocrine pancreas function in young weaners but not in growing-finishing pigs.

A correlation between the exocrine pancreatic function and growth has been previously demonstrated in growing pigs but the data are inconsistent. This was investigated by studying the growth performance of pigs with exocrine pancreatic insufficiency (EPI) at different ages and maintained under similar conditions. Twelve 7 week old (10.5+/-1.3 kg) weaners, and twelve 16 week old (43+/-5 kg) growing-finishing pigs were used in the experiments, and 6 pigs from each group were operated and pancreatic duct-ligated. Starting at 3-5 weeks after the operation, when EPI had developed, weekly recordings of feed consumption and growth were done before, during and after feed supplementation with porcine pancreatin (Creon 10000). In weaner pigs, EPI caused growth arrest while it did not affect the growth of older pigs, as compared to respective un-operated groups of pigs. The daily feed consumption (DFC) was lower in the weaner EPI pigs while it was similar in the growing-finishing EPI-pigs, as compared to un-operated pigs. Feed supplementation with Creon improved the DFC and growth in both the EPI and un-operated pigs. In conclusion, the results showed the importance of the exocrine pancreatic function for growth in weaner pigs, while in older animals it played a minor role in growth. Feed supplementation with pancreatin increased the appetite and ensured an improved feed conversion.

Exocrine pancreatic secretion in pigs fed sow's milk and milk replacer, and its relationship to growth performance.

The objective of this study was to quantify and compare the effects of sow's milk and 2 milk replacer diets (containing clotting or non-clotting protein sources) on exocrine pancreatic secretion, plasma cholecystokinin, and immunoreactive cationic trypsin in pigs. In addition, the relationship between exocrine pancreatic secretion and growth in milk-fed pigs was studied. In a changeover experiment, 9 chronically catheterized pigs of 6.6 +/- 0.19 kg of BW were studied for 3 wk. Pigs were assigned to each of 3 diets. Exocrine pancreatic secretion was measured from the third to the seventh day on each diet. The protein content and trypsin activity of the pancreatic juice were measured. Blood samples were taken at 10 min before and after milk ingestion and were analyzed for cholecystokinin and immunoreactive cationic trypsin. Pancreatic protein and trypsin secretion did not differ between pigs fed sow's milk and those fed milk replacer, but the volume secreted was less for the pigs fed sow's milk (0.75 vs. 1.03 mL x kg(-1) x h(-1); P < 0.01). A postprandial response to milk intake was not observed. The 2 milk replacer diets did not affect exocrine pancreatic secretion differently. The average exocrine pancreatic secretion (volume, 0.94 mL x kg(-1) x h(-1); protein, 4.28 mg x kg(-1) x h(-1); trypsin, 1.65 U x kg(-1) x h(-1)) was intermediate between literature values for suckling and weaned pigs. Plasma cholecystokinin was elevated (approximately 18 pmol x L(-1)) and showed low correlations with the pancreatic secretion traits. Plasma immunoreactive cationic trypsin was not significantly related to any of the pancreatic secretion traits and should therefore not be used as an indicator for exocrine pancreatic function in milk-fed pigs. Exocrine pancreatic secretion varied substantially among individual pigs (protein, 0.22 to 13.98 mg x kg(-1) x h(-1)). Pancreatic protein and trypsin secretion showed a positive, nonlinear relationship with performance traits. It was concluded that neither specific sow's milk ingredients nor the protein source are responsible for a low pancreatic protein secretion in suckling pigs. Exocrine pancreatic secretion was positively correlated with ADG in pigs at an identical milk intake.

Effects of crude red kidney bean lectin (phytohemagglutinin) exposure on performance, health, feeding behavior, and gut maturation of pigs at weaning.

The aim of this study was to obtain information that could help to ease the weaning transition in commercial pig production. Before weaning, phytohemagglutinin (PHA) in the form of a crude preparation of red kidney bean lectin was fed by gavage to 24 crossbred [(Swedish Landrace x Yorkshire) x Hampshire] piglets, whereas 24 control piglets were fed alpha-lactalbumin by gavage, to study the effect on growth, occurrence of postweaning diarrhea, feeding behavior, and some anatomical and physiological traits of the gastrointestinal tract. Within the litter, piglets were randomly assigned to PHA treatment or control and remained in the same pen from the beginning (PHA exposure at 7 d before weaning) until the end of the experiment (14 d post-weaning). Weaning took place at the age of 31 to 34 d. Pigs treated with PHA grew faster (P = 0.013) during the first week postweaning and tended to have lower total diarrhea scores (P = 0.10) than did control pigs. On d 5 after weaning, piglets treated with PHA spent more time eating (P = 0.028) than control pigs. No immunostimulating effect of PHA, measured by plasma immunoglobulin G, could be detected. An increase in the intestinal barrier properties before weaning, as a response to PHA treatment, was demonstrated in intestinal absorption studies using Na-fluorescein and BSA as gavage-fed markers. Less uptake (measured as plasma concentrations) of the marker molecule Na-fluorescein occurred during a 24-h study period, and numerically lower levels of BSA were observed compared with studies in control pigs of the same age. A total of 12 pigs (6 control, 6 PHA-treated) were euthanized on the day of weaning for analyses of gastrointestinal properties. The PHA-treated pigs tended to have a longer total small intestinal length (P = 0.063) than that of the control pigs. The enzyme profile of the jejunal epithelium responded to PHA exposure with a decrease in lactase activity and an increase in maltase and sucrase activities, which is similar to changes normally observed after weaning. No differences were found in the size of the pancreas or in its contents of trypsin and amylase. In conclusion, exposing piglets to crude, red kidney bean lectin for 3 d during the week before weaning led to changes in performance and small intestinal functional properties that would be expected to contribute to a more successful weaning.

Three-day enteral exposure to a red kidney bean lectin preparation enhances the pancreatic response to CCK stimulation in suckling pigs.

BACKGROUND: A reason for the digestive problems that often occur around early weaning in piglets could be that the pancreas is not yet fully developed and the enzymes required for degradation of the solid food are not secreted in enough amounts. OBJECTIVES: The aim of the study was to investigate the possibility of inducing pancreas maturation with enhanced enzyme secretion. METHODS: 10-day-old suckling pigs were gavage fed with a red kidney bean lectin preparation for 3 days, and the pancreatic response to intravenous infusion of CCK-33 was measured in the anaesthetized animals fitted with pancreatic duct catheters. RESULTS: The pancreatic fluid secretion, protein output, and the trypsin and amylase outputs were significantly increased in response to CCK stimulation after the lectin treatment, as compared to those of the control littermates (p < or = 0.05). In addition, the plasma insulin basal levels and those observed during CCK-33 stimulation were lower in the lectin-treated piglets. CONCLUSION: The results suggested that the lectin treatment led to an increase in the capacity for pancreatic enzyme secretion in the suckling piglets. An enhanced pancreatic function might help to ameliorate the problems that may appear in modern pig production which are associated with weaning.

CCK-B receptor antagonist YF476 inhibits pancreatic enzyme secretion at a duodenal level in pigs.

BACKGROUND: To evaluate the mechanisms by which cholecystokinin (CCK) regulates the exocrine pancreas, the role and location of CCK receptors in the pig were investigated using the CCK-B receptor antagonist YF476 and different administration routes of CCK. METHODS: In 11 anaesthetized pigs, catheters were surgically implanted in the pancreatic duct for juice collection, and in the gastric arteries and jugular vein, so that infusions of CCK-33 could be directed to the duodenal/gastric, duodenal/pancreatic or general circulations, respectively. Experiments were performed under control conditions, and after pretreatment by gavage feeding with YF476, using either a single, low dose of 0.3 micromol kg, which would block the CCK-B receptors, or a 1000 times higher dose (300 micromol kg), which would also block the CCK-A receptors. RESULTS: The increase in the pancreatic output of protein and the enzymes trypsin and amylase observed after the infusion of CCK-33 at 13 pmol kg to the duodenum/stomach or duodenum/pancreas was inhibited by pretreatment with YF476 at both dosages. In contrast, the increase in protein and enzyme output after the infusion of a supraphysiological dose of CCK-33 (130 pmol kg) to the general circulation was not affected by pretreatment with low dosage YF476, whereas high dosage YF476 completely inhibited the stimulated secretion. CONCLUSIONS: These data indicate that CCK-33 given locally to the duodenum in doses raising CCK to physiological plasma levels stimulates the pancreatic enzyme secretion via duodenal CCK-B receptors. Supra-physiological doses of CCK-33 to the general circulation appeared to affect the pancreatic enzyme secretion via CCK-A receptors located elsewhere than in the pancreatic and duodenal tissue.

Exogenous leptin controls the development of the small intestine in neonatal piglets.

Leptin, a hormone produced and secreted by adipose tIssue, muscles and stomach, is involved in the regulation of adipose tIssue mass, food intake and body weight in neonatal animals. It is also produced in the mammary glands and secreted into the colostrum and milk. Since leptin receptors are widely distributed in the small intestine mucosa, the aim of the present study was to investigate the effect of exogenous leptin on the development of the small intestine in neonatal piglets. Male neonatal piglets were fed with sow's milk or artificial milk formula. Every 8 h the latter received either vehicle or leptin (2 or 10 microg/kg body weight). The animals were either killed after 6 days of treatment and the small intestine sampled for histology and brush border enzyme activities or were tested for marker molecule (Na-fluorescein and BSA) absorption in vivo. Feeding milk formula slowed the maturation of small intestinal mucosa compared with feeding sow's milk. However, after leptin treatment the length of the small intestine was increased, and intestinal villi length, but not crypt size, was reduced compared with controls. The mitotic index was increased and the percentage of vacuolated enterocytes was reduced in the entire small intestine. Enterocyte brush border protease and lactase activities were reduced in the jejunum. Na-fluorescein marker molecule absorption did not change but that of BSA was reduced 3.8-fold. In conclusion, exogenous leptin administered in physiological doses reversed the maturation of the small intestinal mucosa to the range found in sow-reared piglets.

CCK regulates pancreatic enzyme secretion via short duodenal-pancreatic reflexes in pigs.

BACKGROUND: Different routes of administration of CCK-33 and blockage of CCK-A and muscarinic (m3) receptors are used in this study to evaluate the mechanisms by which cholecystokinin can stimulate the exocrine pancreas. METHODS: The experiment was performed on eight anaesthetized pigs during control conditions and after administration of the CCK-A and m3 receptor antagonists, Tarazepide and 4-DAMP, respectively. Catheters were surgically implanted in the pancreatic duct for juice collection and in the gastric and right gastro-epipoic arteries and in the jugular vein, so that infusions of CCK-33 could be made exclusively to the duodenum/stomach, duodenum/pancreas or general circulation, respectively. RESULTS: Infusion of a low dose of CCK-33 (13 pmol kg(-1)) to the general circulation did not affect pancreatic protein or trypsin output. When the same dose was given directly to the duodenum/stomach or the duodenum/pancreas, pancreatic output increased during both control conditions and after Tarazepide and/or 4-DAMP treatment, though the increase in trypsin output was lower after Tarazepide and/or 4-DAMP blockade. A high dose of CCK-33 (130 pmol kg(-1)) given peripherally stimulated the pancreatic secretion, but this response was totally abolished in Tarazepide and 4-Damp treated animals. CONCLUSIONS: Pancreatic enzyme secretion due to CCK-33 stimulation depends on the presence of short duodenal-pancreatic peptidergic reflexes evoked mainly via low sensitive, probably CCK-B, receptors located in the duodenum/stomach. Pancreatic secretion evoked by peripheral CCK-33 in pharmacological doses was independent of m3 receptors blockade but depended on CCK-A receptors located elsewhere than in the duodenum/pancreas.

Enteral exposure to crude red kidney bean lectin induces maturation of the gut in suckling pigs.

The present investigation characterized the effect of red kidney bean lectin exposure on gut maturation and function in young piglets. Eleven suckling pigs were given by stomach tube a crude red kidney bean lectin preparation (containing about 25% lectin, 400 mg/kg BW) (lectin-treated pigs) at 10, 11, and 12 d of life, and an additional 16 pigs (control pigs) were given saline instead. On the next day, the intestinal absorptive capacity was determined in vivo, and on the 14th d of life the piglets were killed and organs and small intestine samples were collected for analyses and in vitro permeability experiments. The lectin-treated pigs showed an increase in stomach weights and mucosa thickness, whereas no weight effect was found for the small intestine, spleen, liver, or adrenals. Morphometric analyses of the small intestine in lectin-treated pigs showed a decrease in villus heights, an increase in crypt depths and crypt cell mitotic indices, and fewer vacuolated enterocytes per villus and reduced vacuole size. Lectin treatment also resulted in a decrease in the absorption of different-sized marker molecules after gavage feeding, a decrease in intestinal marker permeability, and a change in small intestinal disaccharidase activities, with increased maltase and sucrase activities. The size of the pancreatic acini was also greater in the lectin-treated pigs, but no increases in enzyme content or pancreatic weight could be determined. In addition, the blood plasma levels of cholecystokinin were higher in the lectin-treated than in the control pigs. The results indicate that exposure to crude red kidney bean lectin induces structural and functional maturation of the gut and pancreatic growth in young suckling piglets. This possibility of inducing gut maturation may lead to an improvement in the piglets' ability to adapt to weaning and to an increase in the growth and health of these animals.

Permeability of intestinal mucosa from urinary reservoirs in man and rat.

OBJECTIVE: To evaluate the barrier properties of intestinal mucosa chronically exposed to urine and to evaluate possible differences between ileal and colonic segments used in the reconstruction of the urinary tract. MATERIALS AND METHODS: Mucosal specimens from patients with continent reservoirs with an abdominal stoma, or orthotopic neobladders constructed from colonic segments, were obtained at revisional surgery. Control segments were obtained during right-sided hemicolectomy. In addition, ileal and colonic segments from enterocystoplasties in rats were assessed. The mucosa-to-serosa passage of marker molecules, i.e. (14)C-mannitol, (3)H-glucose, fluorescein isothiocyanate-dextran 4400 and ovalbumin, was measured using modified Ussing diffusion chambers. RESULTS: In man, there were no permeability differences between segments exposed to urine and control segments for any of the marker molecules. In rats, there was less passage of markers in ileal and colonic transplanted segments than in intestinal segments from sham-operated animals. CONCLUSIONS: Intestinal mucosa that has been in chronic contact with urine maintains its barrier function; in the rat model the permeability was even decreased. In addition, there were no detectable differences between ileal and colonic segments in this model.

Intestinal permeability in humans is increased after radiation therapy.

PURPOSE: Irradiation inflicts acute injuries to the intestinal mucosa with rapid apoptosis induction and subsequent reduction in epithelial surface area. It may therefore be assumed that the intestinal barrier function is affected. The aim of this study was to compare the mucosal permeability in irradiated rectum and nonirradiated sigmoid colon from patients subjected to radiation therapy before surgical treatment for rectal cancer. METHODS: Segments from sigmoid colon and rectum obtained from irradiated and nonirradiated patients were stripped from the serosa-muscle layer and mounted in Ussing diffusion chambers. The mucosa-to-serosa passage of the marker molecules 14C-mannitol, fluorescein isothiocyanate-dextran 4,400, and ovalbumin was followed for 120 minutes. RESULTS: The permeability to the markers was size-dependent and increased linearly across time in all specimens. The passage of all markers was increased in irradiated rectum compared with nonirradiated sigmoid colon, whereas in specimens from nonirradiated patients there were no differences between rectum and sigmoid colon. Histologic signs of crypt and mucosal atrophy were found in the irradiated rectal specimens. CONCLUSIONS: Early gastrointestinal complications after radiation therapy may be the result of mucosal atrophy in addition to mucosal damage, with a loss of barrier integrity.

The influence of potato fibre on exocrine pancreatic secretions and on plasma levels of insulin, secretin and cholecystokinin in growing pigs.

The effect of a potato fibre preparation on exocrine pancreatic secretions and on gastrointestinal hormone levels in plasma was studied in three 8 weeks old piglets that were surgically fitted with a jugular vein catheter for blood sampling, a pancreatic duct catheter and a T-shaped duodenal cannula for collection of pancreatic juice. The animals were fed for 2 weeks a control diet (experimental period 1), thereafter for 2 weeks the control diet supplemented with 2% potato fibre (experimental period 2) and for another 2 weeks the control diet again (experimental period 3). Additionally, intraduodenal (i.d.) infusions of the experimental diet, the control diet and potato fibre as well as i.v. infusions of a solution containing cholecystokinin (CCK) and secretin were administered. Potato fibre in the diet evoked in tendency an increase in the volume of secretion of pancreatic juice and a significant increase both in the mean values of the total protein content and total activities of lipase, trypsin and alpha-amylase when compared to the control diet. The i.d. infusion of the control diet, experimental diet and fibre infusate as well as the i.v. administration of the hormone infusate led to a spontaneous secretory response of the exocrine pancreas. Besides gastrointestinal hormones, such as CCK, other factors such as short chain fatty acids may be involved in the regulation of the exocrine pancreas.

Mucosal in vitro permeability in the intestinal tract of the pig, the rat, and man: species- and region-related differences.

BACKGROUND: The barrier properties of the gastrointestinal mucosa may be studied by measuring its permeability to different-sized marker molecules. Owing to difficulties in obtaining human tissue it is, however, often necessary to extrapolate findings from experimental animals to man. The aim of the present study was to compare regional intestinal mucosal permeability in man, the rat, and the pig, using the same marker molecules and in vitro technique. METHODS: Segments from jejunum, ileum, colon, and rectum were mounted in Ussing diffusion chambers, and the mucosa-to-serosa passage of 14C-mannitol, fluorescein isothiocyanate (FITC)-dextran 4,400, alpha-lactalbumin, ovalbumin, and FITC-dextran 70,000 was studied. RESULTS: Irrespective of species or intestinal region an inverse relationship between the molecular weight of the markers and the permeability was seen. The mannitol permeability was higher in the small intestine than in the colon in man, whereas the rat showed a higher permeability in the ileum than in the jejunum and colon. The FITC-dextran 4,400 permeability was higher in all intestinal regions in the rat than in man and the pig. The macromolecules showed low permeability with no regional differences. CONCLUSIONS: The results showed differences between intestinal regions and between species. Permeability data from the pig correlated fairly well with those of man, whereas the rat differed, making it difficult to extrapolate from the rat to man.

Increased colonic permeability in patients with ulcerative colitis: an in vitro study.

BACKGROUND: Colonic permeability was studied in vitro in patients subjected to colectomy because of ulcerative colitis and in control patients undergoing colonic resections for cancer. METHODS: The mucosal layer from fresh colonic segments was stripped and mounted in Ussing diffusion chambers containing modified Krebs buffer solution. The mucosa to serosa passage of the marker molecules 14C-mannitol and ovalbumin was measured for 120 min. RESULTS: Marker passage was significantly increased in colitis patients compared with control patients, irrespective of age, sex, duration of disease, and treatment. Marker passage was further increased in patients with acute colitis. The increased colonic permeability may be explained by inflammation and the resultant loss of mucosal integrity. The increased permeability to ovalbumin implies that permeability to luminal macromolecules, such as bacterial products and other antigenic substances, might be increased in colitis. CONCLUSIONS: The results suggest a derangement of the colonic barrier, as evidenced by an increased mucosal permeability in both chronic and acute colitis.

Effects of systemic and local immunization on alveolar epithelial permeability to protein in the rat.

This study aimed to characterize the effect on alveolar epithelial permeability to protein as a result of immunization when animals were reexposed to antigen. Antigen (bovine serum albumin [BSA]) and a related bystander protein (human serum albumin [HSA]) were intratracheally instilled into lungs of immunized rats and their passages across the alveolar epithelium were measured as serum levels 16 h after instillation. Nonimmunized control rats showed similar passages of both BSA and HSA. In rats intradermally immunized against BSA, BSA was undetectable in serum, whereas serum levels of HSA were markedly increased compared with those in control rats. In rats immunized with BSA intratracheally, serum levels of both BSA and HSA were unchanged compared with those in control rats. Serum titers of specific IgG antibodies (anti-BSA) were measured and were higher in intradermally immunized animals than in intrapulmonary immunized animals, whereas no anti-BSA antibodies were detected in nonimmunized control rats. Anti-BSA antibodies were detected only in lavage fluid from intradermally immunized rats. These findings suggest that presence of specific antibodies locally in the lungs may increase alveolar epithelial permeability to protein. This finding may have clinical implications, e.g., for sensitive asthmatics, since increased nonspecific permeability caused by local immune-related inflammation may result in further allergies.

Enhancing effects of monohexanoin and two other medium-chain glyceride vehicles on intestinal absorption of desmopressin (dDAVP).

The intestinal absorption enhancement of the nonapeptide [Mpa1,D-Arg8]vasopressin (dDAVP) by medium-chain glyceride vehicles was studied using an in vivo rat model. Rats were gavaged with dDAVP formulated with three different lipid vehicles: (1) monohexanoin, (2) mixed monoglycerides, diglycerides and triglycerides of hexanoic acid and (3) monoglycerides, diglycerides and triglycerides of octanoic and decanoic acids, and with saline as control. The marker absorption into blood and urine was followed for 24 hr. All lipid vehicles enhanced the oral bioavailability of dDAVP, but monohexanoin gave the highest increase, approximately 10 times that of control. In contrast to dDAVP, the stable and more lipophilic nonapeptide analog [Mpa1,D-Tyr(ethyl)2,Val4,D-Arg8]oxytocin did not show increased urine recovery when formulated with monohexanoin. A 2-fold increase in urine recovery of the inert low-molecular-weight marker [51Cr]EDTA was observed when formulated with monohexanoin. With use of the fluorescent marker Evans blue formulated with monohexanoin, an elevated accumulation of Evans blue in the mucus layer was observed after incubation in in situ loops. No mucosal damage after lipid vehicle gavage was observed by light microscopic evaluation. Medium-chain glycerides functioned well as oral absorption enhancers of the model peptide dDAVP, and monohexanoin showed the highest enhancement capacity. The mechanisms of this enhancement appear to be related to a protection against luminal dDAVP degradation, mucoadhesive properties of the vehicle and, possibly, an altered epithelial absorption pathway.

Pulmonary delivery of intratracheally instilled and aerosolized cyclosporine A to young and adult rats.

The delivery and pharmacokinetics of cyclosporine A (CyA) given locally to the airways or iv was evaluated in young and adult rats. After intratracheal (i.t.) instillation of saline suspended CyA to adult rats, the CyA plasma levels peaked at 30 min with a bioavailability of 78.1 +/- 6.9%. After the i.t. instillation of CyA with micelles forming surfactant, Cremophor EL, in adult and young rats, the plasma levels peaked at 5 min with a bioavailability of 77.5 +/- 7.2% and 66.3 +/- 4.5%, respectively. The bioavailability of aerosolized CyA was 80.1 +/- 4.1% in adults. Thus, CyA is absorbed by the lungs into the systemic circulation of the rat in high amounts, independent of age and type of delivery system. Long-term treatment with i.t. instillations did not affect body weight gain in young and adult rats, and no histopathological changes were found in the lungs. It is important to emphasize that CyA plasma clearance in young rats was lower and elimination half-life longer than in adults. The slow elimination of CyA in young rats indicated profound pharmacokinetic age differences for this species.

Electrophoretic separation of proteolytic enzymes in pancreatic juice collected with the pouch or catheter method.

CONCLUSION: The results of this study demonstrated that proteolytic enzymes in pancreatic juice from pigs prepared with the pouch method (PM) were nearly fully active or were fully active. When activation with enterokinase was carried out further inactivation and/or breakdown occurred for chymotrypsin C and cathodal trypsin. In addition, some inactivation and/or breakdown of proteolytic enzymes in pancreatic juice occurred during collection of pancreatic juice from PM pigs. METHODS: Samples of pancreatic juice were collected from growing pigs using either the PM or the catheter method (CM). An isolated pouch was prepared where the pancreatic duct enters the duodenum, and three pigs were fitted with a pancreatic pouch re-entrant cannula. Three different pigs had a catheter surgically inserted into the pancreatic duct. Pooled 8-h samples of pancreatic juice were analyzed before and after activation with enterokinase. Chymotrypsin, trypsin, and elastase activities were identified in pancreatic juice after separation by electrophoresis in 1% agarose gels at pH 8.6 using N-acetyl-DL-phenylalanine-beta-naphthyl ester (Ac-Phe-beta ne) as a substrate. RESULTS: This qualitative enzyme assay indicated that a considerable amount of chymotrypsin C, anodal trypsin, chymotrypsins A and B, elastase II, and cathodal trypsin were present in samples of nonactivated pancreatic juice from PM pigs. In contrast, the only active enzymes identified in pancreatic juice from CM pigs were very small amounts of chymotrypsin A and elastase II. The amounts of chymotrypsin C and cathodal trypsin were lower in activated than in nonactivated pancreatic juice from PM pigs. However, there were increases in the amounts of the other enzymes when pancreatic juice from PM pigs was activated. As expected, the activation of pancreatic juice from CM pigs resulted in the measurement of very high amounts of all the proteolytic enzymes. The amounts of anodal trypsin, chymotrypsins A and B, and elastase II were higher in activated pancreatic juice from CM pigs than from PM pigs.

Intestinal inflammation and barrier function in HLA-B27/beta 2-microglobulin transgenic rats.

BACKGROUND: Since intestinal inflammation is correlated with impaired barrier functions, transgenic HLA-B27/human beta 2-microglobulin rats that spontaneously develop intestinal inflammation were used to investigate whether onset of inflammation or impaired barrier function was the initial event. METHODS: During the age period of 9-14 weeks, transgenic and non-transgenic (control) rats were gavaged weekly with the marker molecules, 51Cr-ethylenediaminetetraacetic acid, 1-deamino-8-D-arginine vasopressin, and albumin, which were quantified in blood or urine. RESULTS: At 12 weeks of age the first signs of inflammation appeared with decreased body weight gain, decreased urine production, and onset of diarrhea. By 14-15 weeks of age all transgenic rats had developed intestinal inflammation, as confirmed by histology and increased myeloperoxidase content, whereas no inflammation was observed in controls. Intestinal passage of the markers did, however, not differ between transgenic and control rats over the studied period. CONCLUSIONS: The results suggest that intestinal inflammation precedes altered intestinal barrier function in this inflammation model.