Trypsin Inhibitor Content Research Articles

Measurement of blood and plasma coagulation time using free oscillating rheometry

An assay based on free oscillating rheometry to measure the activity of coagulation factors is described. The method can be used in blood and plasma and is particularly suitable for screening and monitoring coagulation disturbances in point-of-care testing (POCT) in environments where quick analysis with minimal preanalytical work is needed. In this study the endpoint as clotting onset time (COT) is determined by a deviation from initial viscoelastic properties of an oscillating sample. The model system entails the clotting of citrated blood or plasma clotting by repletion of Ca 2+ . COT was shown to give a dose-dependent response to added thrombin and to be resistant to high concentrations of corn trypsin inhibitor, indicating measurement of the tissue-factor-dependent pathway of coagulation activation. COT in recalcified blood and plasma covariated with prothrombin time (PT) according to Owren, and activated partial thromboplastin time (aPTT). The technique and instrument used proved to be quick and easy to handle, and suitable for POCT as well as for examinations in the laboratory.

Selectivity of IMAC columns in trypsin inhibitor purification.

The properties of an adsorbent and the parameters in an adsorption process affect the resolution of chromatographic purifications. This is reflected in the elution profile, which shows the relative affinity of different proteins for a specific adsorbent. In the work presented here, elution profiles for trypsin inhibitor were used to study the effects of the concentration of trypsin inhibitor, ionic strength of the protein solution, slope of the elution gradient, and the regeneration treatment of the chromatography column on the selectivity of the adsorbent Cellufine Chelate-Cu(II)(ida). Cytochrome c was used as a reference protein. Variations in the concentrations of trypsin inhibitor and in the ionic strength of the buffered solution did not have any effects on the elution profile. On the other hand, changes in the slope of the pH gradient used for elution caused shifting of the elution peaks toward lower values of the elution volume, resulting in the best strategy to modify the elution profile of the system. Finally, using a constant slope pH gradient of elution, the variation of the selectivity of the adsorbent for trypsin inhibitor when subjected to cleaning treatments with 0.5 N NaOH was studied. Appropriate cleaning practices used in industry were followed. The adsorbent showed only a slight tendency for resolution loss in the order of 2 x 10(-4) days(-1). The results presented here show a good stability of the adsorbent when compared to other biospecific adsorbents commonly used.

Effect of various domestic processing methods on antinutrients and in vitro protein and starch digestibility of two indigenous varieties of Indian tribal pulse, Mucuna pruriens Var. utilis.

The effect of various domestic processing methods on antinutrients and starch fractions and in vitro protein and starch digestibilities of white and black varieties of Mucuna pruriens var. utilis was studied. Cooking or autoclaving of both raw seeds and presoaked seeds in different solutions (water, tamarind extract, sodium bicarbonate, and citric acid) significantly (p < 0.05) reduced the content of total phenolics, phytic acid, trypsin inhibitor and chymotrypsin inhibitor activities, and L-dopa compared to soaking or dry heating techniques. The germination processes (24 and 48 h) were also effective in the reduction of various antinutrients, although this reduction appeared to be more pronounced in a prolonged period of germination (72 h). Water soaking followed by dehusking was found to be ineffective in the reduction of trypsin and chymotrypsin inhibitor activities in both varieties. All of the treatments were effective in significantly (p < 0.05) reducing the resistant starch content in the presently investigated samples. Cooking as well as autoclaving brought about a more significant (p < 0.05) improvement in the digestibility of protein and starch compared to germination and dry heat treatment. Moreover, among the different processing techniques, soaking in sodium bicarbonate solution followed by cooking (29.6-34.8%) or autoclaving (33.0-37.2%) seemed to be the best method for improving starch digestibility.

Plant density and nutrient availability constrain constitutive and wound-induced expression of trypsin inhibitors in Brassica napus.

We investigated the effects of plant density on plant size, leaf total soluble protein content, and constitutive and wound-induced levels of proteinaceous trypsin inhibitors in pot-grown Brassica napus seedlings in two greenhouse studies. We manipulated plant density by varying the number of intraspecific neighbors surrounding a target plant in the center of each pot. In general, constitutive and induced levels of trypsin inhibitors were significantly reduced by competition in a density-dependent manner, to the extent that induction was greatly reduced or abolished in target plants surrounded by six neighbors. To investigate whether the effects of plant density on inhibitor production were mediated by nutrient availability, we manipulated the concentration of a complete fertilizer applied to target plants surrounded by six neighbors in two greenhouse studies. In general, constitutive and wound-induced levels of inhibitors in plants surrounded by six neighbors were increased by nutrient addition in a dose-dependent manner, such that wound-induction was completely restored in competing plants under conditions of high nutrient availability. Leaf total soluble protein content, measured only in the second trial of each experiment, was not affected by any of the treatments. The effects of plant density, nutrient addition, and wounding on inhibitor levels in all experiments were independent of their effects on above-ground plant size at the time of wounding. Overall, our results suggest that decreasing nutrient availability mediates the density-dependent reductions in inhibitor levels in B. napus seedings.

Trypsinogen-1, -2 and tumour-associated trypsin-inhibitor in bile and biliary tract tissues from patients with biliary tract diseases and pancreatic carcinomas

The bile concentrations of trypsinogen-1, -2 and tumour-associated trypsininhibitor (TATI) were determined in 23 patients with benign biliary tract disease, two with biliary tract cancer, and in 15 with pancreatic cancer. We also examined the trypsinogen and TATI expression by immunohistochemistry in tissue specimens from biliary tract cancer and non-neoplastic extrahepatic biliary tract. High levels of trypsinogen-1, trypsinogen-2, and TATI occur in bile of most patients. In contrast to the trypsinogens, the levels of TATI were significantly higher in patients with malignant disease than in those with benign diseases (p=0.04). There was no significant correlation between trypsinogen-2 and amylase (r= 0.13, p=0.40), indicating that the occurrence of trypsinogen in bile is not a result of reflux of pancreatic fluid into the bile duct. Immunohistochemically, trypsinogen-2 was detected in five and TATI in 12 out of 15 non-neoplastic biliary tract specimens, and in four and seven out of 11 cholangiocarcinomas, respectively. High concentrations of trypsinogen-1, trypsinogen-2 and TATI occur in the bile of patients with non-neoplastic and malignant biliary tract disease and in patients with pancreatic cancer. At least part of the trypsinogen-2 and TATI found in bile appears to be derived from the biliary epithelium itself.

Urinary trypsin inhibitor concentration can predict the immunological insult of chemotherapy and complications after bone marrow transplantation.

Urinary trypsin inhibitor has attracted attention as an index of the systemic inflammatory response syndrome. In this study, the urine concentration of trypsin inhibitor was measured to compare the immunological insult of conventional chemotherapy and conditioning chemotherapy for bone marrow transplantation. We also investigated whether urinary trypsin inhibitor was a useful index of the complications and outcome of bone marrow transplantation. Urinary trypsin inhibitor concentration was determined before chemotherapy, on the day after finishing chemotherapy (day 0 of transplantation), and during recovery of the white cell count, in 17 patients (seven receiving conventional chemotherapy and 10 receiving conditioning for bone marrow transplantation). Urinary trypsin inhibitor concentrations were significantly higher after conditioning for bone marrow transplantation than after conventional chemotherapy (P < 0.001), indicating that conditioning was more invasive. After bone marrow transplantation, the incidence of severe complications and the mortality rate were higher in patients whose urinary trypsin inhibitor concentrations rose during recovery of the white cell count. Comparison of urinary trypsin inhibitor concentrations suggested that conditioning for bone marrow transplantation was more invasive than conventional chemotherapy. This study also suggested that the urine concentration of trypsin inhibitor could be useful for predicting the risk of complications and outcome of bone marrow transplantation.

Effects of dry matter content on trypsin inhibitors and urease activity in heat treated soya beans fed to weaned piglets

A nutrition study was conducted to evaluate the growth response of weaned piglets fed diets containing soya beans that had been processed into protein supplements at two different levels of dry matter (DM) and temperature. Four diets contained protein supplements prepared from whole full-fat soya beans equilibrated at 800 or 900 g kg −1 DM prior to being heated to 110 or 125°C. An additional diet contained a protein supplement prepared from raw whole full-fat soya beans at 900 g kg −1 DM; i.e. an unheated soya bean protein supplement. The experimental control diet was supplemented with a solvent extracted commercially processed soya bean meal (900 g kg −1 DM) containing 480 g crude protein kg −1. Soya beans at 900 g kg −1 DM prior to heat treatment at 110°C produced protein supplements, after heat treatment, that had higher residual levels of trypsin inhibitors and urease activity than measured in soya beans at 800 g kg −1 DM prior to the same heat treatment. The moisture content of soya beans prior to heat treatment affected the level of heat necessary to lower values for trypsin inhibitors and urease activity. Soya beans at 800 g kg −1 DM prior to heating at 110°C, produced a protein supplement with similar residual concentrations of trypsin inhibitors and urease activity to soya beans at 900 g kg −1 DM prior to heating at 125°C. This observation suggested that the soya beans with higher moisture content required lower heat energy to inactivate trypsin inhibitors and urease. The pen unit response of piglets fed the diet containing the soya bean protein supplement prepared from soybeans processed at 900 g kg −1 DM and heated to 110°C was not improved when compared to piglets fed the diet containing the unheated soya bean protein supplement. Soya beans at 800 or 900 g kg −1 DM prior to heating to 110°C, or soybeans at 900 g kg −1 DM heated to 125°C, produced protein supplements that were inadequately heat processed as indicated by the values for residual trypsin inhibitors and urease activity, and, the depressed pen unit response of piglets when compared to those fed the control diet. In contrast, piglets fed the protein supplement prepared from soya beans at 800 g kg −1 DM prior to heating to 125°C, displayed an average daily feed intake and feed-to-gain ratio that did not differ significantly from piglets fed the control diet. These data indicate that when whole full-fat soya beans were processed by the dry roasting method, their initial DM content of 800 or 900 g kg −1 affected the processing temperature necessary to denature or otherwise inactivate inherent trypsin inhibitors and urease activity.

Chemical Composition and Antinutrient Content of three Lupinus Species from Jalisco, Mexico

In this study, the proximal chemical analysis and contents of antinutritional factors (lectins, antitrypsin activity, cyanogenic glycosides, alkaloids, phytates and α -galactosides) of Lupinus exaltatus, L. reflexus, and L. mexicanus seeds were determined. The seeds of these lupins comprised 384.1, 388.0, 367.0 g/kg protein, which contained all the essential amino acids for human beings except glutamine and asparagine in their seed protein. OnlyL. exaltatus was free from lectins. Trypsin inhibitor concentrations ranged from 1.12 to 2.05 TIU/mg. Cyanogenic glycosides were present at low concentrations in the studied lupins. Total alkaloid content ranged from 14.0 to 44.0 g/kg and phytate content ranged from 11.1 to 1.856 g/kg. The main α -galactosides found in seeds were raffinose, stachyose, verbacose and ajucose. Stachyose was the predominant sugar in the studied species. Therefore, these wild lupins could be considered a good source of protein after a suitable reduction in the content of alkaloids.

Effects of extrusion cooking on some functional properties of soy-sweet potato mixtures--a response surface analysis.

Mixtures of defatted soy flour and sweet potatoes were extruded at 0:100, 25:75, 50:50, 75:25 and 100:0 ratios in an Insta-Pro 600 single screw extruder operated at 180, 200, 220 rpm. Oil (3-5%) was added into the mix before extrusion. A 5 x 32 fractional factorial central composite response surface design was adopted for investigating the variables: feed composition, % oil, and screw rotation speed (RPM). Percent oil and screw speed were randomly investigated at three levels, while feed composition was randomly investigated at five levels. Effect of extrusion cooking on functional properties: bulk density, expansion ratio, water absorption and solubility indices and trypsin inhibitor were assessed. Results showed that feed composition and screw speed had strong influences on the process. The effect of adding oil, into defatted soybean significantly (p < 0.05) affected only trypsin inhibitor content. High extrusion temperature (> 130 degrees C), and accompanying high shear, were the result of the non-pasting nature of the extrudate. Therefore the products may have limited uses.

Detectable concentrations of urinary trypsin inhibitor in cerebrospinal fluid.

Urinary trypsin inhibitor (UTI) is a physiological protease inhibitor produced in the liver and excreted into urine. Urinary trypsin inhibitor-like antigen has been demonstrated on glial cells in the brain. This study measured cerebrospinal UTI levels in various conditions. Seven subjects in each of the following groups were studied: patients undergoing spinal anesthesia for minor surgery, spinal anesthesia for cesarean section, removal of meningioma, or at postoperative day 3 after ruptured intracranial aneurysm clipping. Cerebrospinal fluid was collected from a spinal needle, a needle puncturing the sylvian fissure, or a drainage tube from the optical carotid cistern. Urine was collected from a urinary catheter. Cerebrospinal and urinary UTI concentrations were measured by radioimmunoassay, and the urinary UTI concentration was divided by urinary creatinine concentration to give the systemic UTI concentration. The cerebrospinal UTI concentration in the brain tumor and postoperative state groups was significantly higher than in the spinal anesthesia and cesarean section groups. The systemic UTI concentration in the cesarean section and postoperative state groups was significantly higher than in the spinal anesthesia and brain tumor groups. The present results demonstrate that UTI can be detected in the cerebrospinal fluid, and suggest that cerebrospinal UTI increases in patients with a brain tumor or inflammation and is not affected by systemic UTI.

The trypsin inhibitor content of 61 wild edible plant foods of Niger.

In the western Sahel and many other regions of sub-Saharan Africa, wild edible plants contribute significantly to human diets, not only during periods when cereal staples are scarce, but also when they are readily available. Although there have been published reports regarding the nutrient contents of these plant foods, little attention has been devoted to their content of antinutrients such as calcium chelators and inhibitors of the pancreas-derived proteases, trypsin and chymotrypsin, which are required for the efficient digestion and absorption of dietary proteins. In this study, aqueous extracts of 61 different leaves, seeds, fruits and flowers of edible plants gathered in the Republic of Niger were analyzed for their content of trypsin inhibitory substances using alpha-N-benzoyl-DL-arginine-p-nitroanilide as the substrate and bovine trypsin as the enzyme source. Twelve of these plant foods contained more antitrypsin activity than soybeans (1.34-8.18 vs. 1.32 microg trypsin inhibited/mg dry weight). Boiling for 3 min did not inactivate the antitrypsin activity in most of the plant extracts. These data confirm that more than half of the wild edible plant foods widely consumed by various populations who inhabit the western Sahel contain significant quantities of heat-stable trypsin inhibitor that could possibly compromise the bioavailability of proteins present in the diets of these populations.

Studies on the biological responses of rats to seed trypsin inhibitors using near-isogenic lines ofPisum sativum L (pea)

In order to determine the true antinutritional status of pea seed trypsin inhibitor (TI) proteins, pea lines are being produced that are near-isogenic except for the genetic locus, Tri, containing the TI structural genes. These lines are based on selection from the progeny of a cross between lines showing quantitative variation in TI content, as well as TI isoform and gene polymorphisms that serve as markers. Chemical analyses revealed that the composition of seeds from lines of each near-isogenic pair was extremely similar, except for a more than five-fold difference in TI content. Such lines provide material that is superior to the diverse lines previously used for nutritional assessment of pea TI. The specific biological effects of pea TI were studied by including the near-isogenic lines in standardised rat diets. The results indicated that TI content was correlated with a significant negative effect on protein digestibility and biological value. The difference in TI content of the pea seeds was reflected in the relative activity of pancreatic chymotrypsin whereas the activities of trypsin, lipase and amylase were less clearly affected. © 1999 Society of Chemical Industry

Studies on the biological responses of rats to seed trypsin inhibitors using near‐isogenic lines of Pisum sativum L (pea)

In order to determine the true antinutritional status of pea seed trypsin inhibitor (TI) proteins, pea lines are being produced that are near-isogenic except for the genetic locus, Tri, containing the TI structural genes. These lines are based on selection from the progeny of a cross between lines showing quantitative variation in TI content, as well as TI isoform and gene polymorphisms that serve as markers. Chemical analyses revealed that the composition of seeds from lines of each near-isogenic pair was extremely similar, except for a more than five-fold difference in TI content. Such lines provide material that is superior to the diverse lines previously used for nutritional assessment of pea TI. The specific biological effects of pea TI were studied by including the near-isogenic lines in standardised rat diets. The results indicated that TI content was correlated with a significant negative effect on protein digestibility and biological value. The difference in TI content of the pea seeds was reflected in the relative activity of pancreatic chymotrypsin whereas the activities of trypsin, lipase and amylase were less clearly affected. © 1999 Society of Chemical Industry

Changes in flax ( Linum usitatissmum) seed nitrogenous compounds during germination

The changes in the nitrogenous compounds of flaxseed during an 8-day germination period were studied. The dry matter content of the seeds was reduced by 35% at the end of the germination. During the germination period, a relatively small decrease was observed in total nitrogen content, but there was an increase in the content of non-protein nitrogen from 9 to 33.5% of the total amount. An increase in the total content of free amino acids was also observed. Among individual amino acids, glutamine showed a marked change during the germination period indicating that it is the favoured amide donor in the developing flax seedlings. An increase in the water-soluble protein and a decrease in the salt-soluble protein fractions was also observed. The content of polyamines, namely agmatine, spermidine and putrescine, which are important in controlling cellular metabolism and growth, was also increased during the germination period. After day 8 of germination, the contents of cyanogenic glycosides, linustatin and neolinustatin in the seeds were reduced by 40 and 70%, respectively. Trypsin inhibitor content was very low in flaxseed and only trace amounts were present after 8 days of germination.

Inhibition of plant-pathogenic fungi by a corn trypsin inhibitor overexpressed in Escherichia coli.

The cDNA of a 14-kDa trypsin inhibitor (TI) from corn was subcloned into an Escherichia coli overexpression vector. The overexpressed TI was purified based on its insolubility in urea and then refolded into the active form in vitro. This recombinant TI inhibited both conidium germination and hyphal growth of all nine plant pathogenic fungi studied, including Aspergillus flavus, Aspergillus parasiticus, and Fusarium moniliforme. The calculated 50% inhibitory concentration of TI for conidium germination ranged from 70 to more than 300 microgram/ml, and that for fungal growth ranged from 33 to 124 microgram/ml depending on the fungal species. It also inhibited A. flavus and F. moniliforme simultaneously when they were tested together. The results suggest that the corn 14-kDa TI may function in host resistance against a variety of fungal pathogens of crops.

Genotype � environment influence on cowpea (Vigna unguiculata (L) walp) antinutritional factors: 1 - trypsin inhibitors, tannins, phytic acid and haemagglutinin

The relative influence of genotype, environment and genotype×environment effects on four antinutritional factors (g kg−1) of importance in cowpea were studied using 15 local and improved cowpea genotypes grown in 12 environments, comprising three locations over four seasons per location. The locations Ago-Iwoye (6°58′N4°00′E), Mokwa (9°17′N5°04E) and Kano (12°00′N8°31′E) were representatives of the major agroecological zones where cowpeas are produced. Genotypes effects were strongest in controlling trypsin inhibitor content, while the environment was the major source of variation for tannins, haemagglutinin and phytic acid contents. Thus, the variability in the levels of these antinutritional factors in cowpea seeds depends largely on the environment where they are grown. This implies that a cowpea genotype grown and consumed safely in an environment can be poisonous when grown and consumed in another environment. Genotype×environment effects were significant for tannins, haemagglutinins and trypsin inhibitor contents. Correlation coefficients (pooled data) from the three locations indicated that trypsin inhibitor was positively correlated to phytic acid (r=0.59, 0.001<P<0.05) and haemagglutinins (r=0.64, 0.001<P<0.05) but negatively correlated to tannin contents ( r=−0.79, 0.001<P<0.05). © 1999 Society of Chemical Industry

Nutrient and antinutrient composition of Amaranthus muricatus.

The chemical and nutritional composition of the aerial parts of Amaranthus muricatus was assessed. This is a wild plant growing well in arid and poor soil, which are not suitable for cereals. The material under study was boiled, dried and ground and then evaluated for chemical, nutritional and antinutritional properties. Protein concentration of the amaranth flour was 15.74 g/100 g and total dietary fiber content was high (53.81 g/100 g), with 79% coming from insoluble dietary fiber. The calcium content was 1533 mg/100 g and the Ca/P ratio (5.94) was markedly higher than the value suggested in the 1989 Recommended Dietary Allowances. Fifty-nine percent of the fatty acids present were unsaturated with linoleic acid accounting for 40% of the total fatty acid content. Nitrate, hemagglutinin, trypsin inhibitor and saponin contents were within the non-critical values. Net protein utilization (NPU), true digestibility (tD) and biological value (BV) measurements indicated a nitrogen gain of 74% that of casein. The results of this study indicate a nutritive potential for the A. muricatus leaves, therefore, domestication of this plant is suggested along with assessment of its chemical and nutritional properties.

Elevated Serum Inhibin and Tumor-Associated Trypsin Inhibitor Concentrations in a Young Woman with Dysgerminoma of the Ovary

We describe a patient with dysgerminoma who had elevated serum inhibin, tumor-associated trypsin inhibitor (TATI), and CA 125 concentrations, which increased progressively during follow-up of the advancing disease. Inhibin levels correlated closely with disease behavior. In contrast to inhibin, serum TATI and CA 125 failed to reveal the presence of silent disease.

Serum Carcinoembryonic Antigen, Cancer Antigen 125, Cancer Antigen 15-3, Squamous Cell Carcinoma, and Tumor-associated Trypsin Inhibitor Concentrations during Healthy Pregnancy

In the management of cancer patients, tumor-associated antigens are measured in serum as noninvasive tests for relapse detection [reviewed in Ref. (1) ]. The tests have limited specificity because the serum concentrations of the antigens can be affected by pathophysiological conditions such as renal insufficiency, hepatic diseases, and inflammation (2) . The temporal changes of tumor markers during pregnancy are not well documented. Although some have reported values grouped by trimester of pregnancy (3) , longitudinal studies are rare. We have studied the tumor markers CEA (carcinoembryonic antigen), CA (cancer antigen) 125, CA 15-3, SCC (squamous cell carcinoma), and TATI (tumor-associated trypsin inhibitor), which are widely used in gynecological tumors. CEA is expressed in many malignancies and is useful for the monitoring of colon, lung, and breast cancers (4) . CA 125 and CA 15-3 belong to the family of carbohydrate antigens also named mucins (1) . CA 125, which was identified by a monoclonal antibody raised to an ovarian carcinoma cell line (OC125), is used for the follow-up of nonmucinous ovarian cancer (1) . It displays increased seric concentrations in cases of serous effusions (2) . CA 15-3, which was identified by two monoclonal antibodies 115D8 and DF3, is the most widely used tumor marker for breast carcinoma (4) . SCC (or Ta-4) is a tumor-associated antigen isolated from squamous cell carcinoma of uterine cervix (5) and is used for the follow-up of epidermoid tumors (cervical uterine carcinoma or epidermoid lung cancer) (6) . TATI was isolated from the urine of an ovarian cancer patient (7) . It is used for the follow-up of mucinous ovarian cancer (8) and other malignancies including cancers from the urogenital …

DOSE-DEPENDENT PANCREATOTROPHIC EFFECT OF CHOLECYSTOKININ-OCTAPEPTIDE IN THE RAT: THE INFLUENCE OF STARVATION

Cholecystokinin (CCK) and its analogues are known to exert trophic effects on the exocrine pancreas, whereas at high doses, they produce pancreatic injury. This study was carried out to study the effect of starvation on the dose-dependent pancreatotrophic effect of CCK-8 in rats. Normal or fasted rats were treated with CCK-8 doses ranging from 0.5 to 32 and 0.5 to 8 μg kg−1, respectively, twice daily for 5 days. Pancreatic size, protein, DNA, secretory enzyme and trypsin inhibitor (PSTI) contents as well as histology were examined. In normal rats, CCK-8 increased the pancreatic content of protein, amylase, serine proteases and PSTI with maximum values between doses of 2 and 16 μg kg−1. The dose of 32 μg kg−1, however, yielded less trophic responses. Given to fasted rats, CCK-8 increased the weight as well as protein and secretory enzyme contents of the pancreas with maximum values between doses of 1 and 4 μg kg−1. The first dose supramaximum for the trophic responses was as low as 8 μg kg−1. Histology revealed necroinflammatory damage (acinar cell vacuolization, focal cell necrosis) in the exocrine pancreas at supramaximum doses of CCK-8 in both groups. Cell necroses and vacuolization were less but present even at doses optimum for trophism and exhibited dependence on both the dose of CCK-8 and nutrition. In either the normal or fasted animals, the periinsular acini were relatively less affected by the toxic effects of CCK-8 than the teleinsular ones. The results indicate that starvation makes the exocrine pancreas more sensitive to necroinflammatory effects of CCK-8. The relative protection seen in periinsular acini suggests a modulatory influence of islet hormones on development of CCK-induced acinar cell injury.