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alpha-Amylase of human pure pancreatic juice: effects of pancreatic disease and the occurrence of variant forms in pancreatic juice from healthy volunteers.

Pure pancreatic juice (PPJ) from healthy human volunteers and from patients with pancreatic or liver disease was subjected to isoelectric focussing (IEF) and assayed for alpha-amylase activity. In PPJ from most normals, a single predominant form of amylase was found, comprising congruent to 83% of the total activity recovered, and having pIapp congruent to pH 6.8. In PPJ from six normals, variant principal forms of amylase were found at pH congruent to 6.4 or pH congruent to 7.3, in addition to the peak at pH 6.8. IEF patterns of PPJ from individuals with pancreatic or liver disease were indisquishable from patterns obtained with PPJ from the control group of healthy volunteers.

Amylases

Trypsin and lactoferrin levels in pure pancreatic juice in patients with pancreatic disease.

Levels of immunoreactive trypsin were measured in pure pancreatic juice obtained endoscopically from 44 patients with suspected pancreatic disease. Patients with pancreatic cancer all had low trypsin concentrations (median 3.6 micrograms/ml, range 0.6--12.0), but those with chronic pancreatitis had very variable levels (median 14.2 micrograms/ml, range 3.2--76.8), showing a considerable overlap with patients without pancreatic disease (median 37.1 micrograms/ml, range 10.4--66.0). When levels of lactoferrin in pancreatic juice were measured, all patients with chronic pancreatitis were found to have much higher levels (all greater than 900 ng/ml) than control subjects or patients with pancreatic cancer (all less than 400 ng/ml). The combined measurement of trypsin and lactoferrin in pure pancreatic juice appeared to be more promising than any other currently available test for the separation of patients with pancreatic cancer from those with chronic pancreatitis.

Chronic Disease

Radioimmunoassay of lactoferrin in pancreatic juice as a test for pancreatic diseases.

Lactoferrin, a protein present in pancreatic juice and other exocrine secretions, was measured by radioimmunoassay in pure pancreatic juice obtained by endoscopic cannulation of the pancreatic duct. Lactoferrin concentrations were high in pancreatic juice from patients with chronic pancreatitis, but they were considerably lower in juice from control subjects and patients with carcinoma of the pancreas. The measurement of lactoferrin concentrations in pure pancreatic juice may be useful in the diagnosis of pancreatic diseases.

Chronic Disease

The phospholipase activity of sheep pancreatic juice.

Sheep pancreatic juice was found to contain at least two enzymes which hydrolysed biliary lecithin. One enzyme was heat and acid labile and hydrolysed the fatty acid from position 1 (phospholipase A1); the other was heat and acid stable hydrolysing the fatty acid at position 2 (phospholipase A2). Lysophospholipase activity was also present. The phospholipases were active at pH values greater than 4.2, and would therefore function in the acid conditions (pH 3-6) of the sheep small intestine. The activity of the pancreatic phospholipases, and A2 in particular, was dramatically stimulated by the presence of the secretions of Brunner's glands which could be important in accelerating the hydrolysis of biliary lecithin in the lumen of the intestine. Phospholipase A1 was sensitive to acid in the range pH 2.5-3.5 and could therefore be partially inactivated by abomasal digesta; but phospholipase A2 was resistent to acid treatment.

Animals

Non-parallel secretion of pancreatic enzymes. Effect of parenteral amino acid administration and intestinal reinfusion of bilio-pancreatic juice.

Chronic drainage of pancreatic juice in rats results in a non-parallel secretion of digestive enzymes. The concentration of trypsinogen, chymotrypsinogen, procarboxypeptidase B and lipase increases continuously whereas that of amylase drops. An amino acid mixture given intravenously does not affect this dissociation. Reinfusion of bilio-pancreatic juice into the duodenum on the contrary influences the relative enzyme release: the increase in concentration of the three proteolytic enzymes and of lipase is less pronounced over the 72-hour study period; amylase concentration remains unaffected. In the absence of an intravenous amino acid feeding the pancreatic protein output is more than halved and a negative nitrogen balance is observed. Reinfusion of bilio-pancreatic juice does not alter the total protein output. This report confirms the persistance of a dissociation in the rate of enzyme release in rats with chronic pancreatic fistula despite sufficient protein administration and intestinal reintroduction of bilio-pancreatic juice.

Amino Acids

Trypsinogen variants in pancreatic juice of healthy volunteers, chronic alcoholics, and patients with pancreatitis and cancer of the pancreas.

Polyacrylamide gel electrophoresis of pure pancreatic juice from 14 healthy normal subjects, 11 chronic alcoholics without detectable pancreatic disease, 15 patients with pancreatitis, and two with cancer of the pancreas consistently demonstrated the presence of two variants of trypsinogen with different electrophoretic mobilities. In healthy normal subjects the proportion of cationic to anionic trypsinogen was invariably greater than 1 and averaged about 2. In chronic alcoholics, patients with pancreatitis or cancer of the pancreas, this ratio, with a single exception, was below one and averaged about 0.45. The extraordinary consistency of these findings suggests that the quantitative relationship between cationic and anionic trypsinogen in human pancreatic juice may be a very sensitive indicator of incipient or existing pancreatic pathology. The most acceptable explanation for the reversal of the normal zymogen ratio in pancreatic disease is a selective increase in the synthesis of the anionic variant relative to that of the cationic species. Total trypsinogen concentrations differed widely from one another in the three patient groups, but the ratio of cationic to anionic trypsinogen exhibited little change and remained below 1. Our results also demonstrate for the first time a specific effect of chronic alcohol abuse on the secretory profile of a pancreatic enzyme in human subjects. A newly discovered minor, trypsinogen-like component of human pancreatic juice was found to be significantly increased in pancreatic juice of chronic alcoholics, decreased in pancreatic secretions of patients with pancreatitis, and barely detectable in those of two patients with cancer of the pancreas.

Adult

Pure pancreatic juice studies in normal subjects and patients with chronic pancreatitis.

Pure pancreatic juice was obtained from within the pancreatic duct in 54 patients after endoscopic cannulation of the papilla of Vater. In all 20 normal subjects there was a brisk response to intravenous injections of GIH secretin in small dosage (1 and 4 CU). Peak bicarbonate concentrations occurred after a 4 CU stimulus, whereas volumes, and bicarbonate and protein outputs were greatest after 70 CU. Total protein and amylase concentrations were highest in the first specimens collected from each patient, and fell rapidly after stimulation. Plateau levels for all indices were achieved 10-20 minutes after starting infusions of secretin and pancreozymin. When normal patients and those with chronic pancreatitis were compared, there was considerable overlap in all indices (volume, bicarbonate and total protein concentrations) after bolus injections of secretin. Most patients with chronic pancreatitis achieved a peak bicarbonate concentration in excess of 100 mmol/l. The median concentrations were not significantly different from normal after any dose of secretin when pooled 10 minute samples were analysed. However there were significant differences in peak bicarbonate concentrations (after 1 and 4 CU, but not after 70 CU), when one minute samples were compared. There were also statistically significant differences in the median 10 minute responses for volume after 1 and 70 CU, for bicarbonate output after 1, 4, and 70 CU, and for protein output after 70 CU. The results of juice studies in patients believed to have early chronic pancreatitis did not differ significantly from those in normal subjects or those with chronic pancreatitis. Endoscopic duct cannulation cannot guarantee complete recovery of pancreatic secretions, and measurements of volume and output may be inaccurate. When standard biochemical indices are used, the diagnostic role of pure juice studies is limited; further research may reveal more specific disease markers.

Bicarbonates

Analysis of pure pancreatic juice in patients with chronic alcoholism.

To elucidate early biochemical changes of pancreatic juice and their reversibility in chronic alcoholics, pure pancreatic juice was collected from 23 chronic alcoholics by endoscopic retrograde catheterization of the papilla. Samples were collected at 1 minute intervals for 20 minutes after intravenous injection of secretin (Eisai, 1 U/kg) and for 10 minutes after CCK-PZ injection (Boots, 1 U/kg). Volume, bicarbonate concentration, protein concentration and three hydrolases were determined. Following results were obtained. (1) Five patients showed hypersecretory state. Four of the five patients showed hyperconcentration of protein. (2) Seventeen patients showed hyposecretory state. Lipase secretion was most frequently affected (94%). Maximal bicarbonate concentration was the next to be affected (82%). Amylase and chymotrypsinogen secretion were less frequently affected (65%). Flow rate was least frequently affected (24%). (3) It was suggested that exocrine dysfunction in chronic alcoholics is reversible in an early stage and that sequence of events with advancement of the stage is hypersecretion, hyperconcentration of protein, normalization of water secretion with a decrease in lipase secretion and maximal bicarbonate concentration, a decrease in amylase and chymotrypsinogen output, and finally a decrease in flow rate.

Adult

[Effect of intra-duodenal administration of HCl and NaHCO3 on the secretion and content of sodium, potassium and calcium in the pancreatic juice and bile of sheep].

The experiments were carried out on 3 sheep with a Harrison type cannula inserted into the funds of the gall-bladder enabling to obtain bile and with a Herrera type cannula fixed in the duodenum enabling to sample pancreatic juice. Administration of 3 mEq HCl in 50 ml of solution to the duodenum increased the volume of secreted bile from 6,04 ml to 8,11 ml/15 min with no essential changes in the volume of pancreatic juice. Administration of 15 mEq NaHCO3 in 50 ml of solution decreased the volume of secreted bile from 6.73 ml to 4,15 ml/15 min and decreased pancreatic juice from 5,46 ml to 4.72 ml/15 min. Intravenous administration of secret increased the volume of both pancreatic juice and bile. Administration of HCl and NaHCO3 solution to the duodenum did not cause changes in the concentration and secretion of sodium, potassium and calcium in pancreatic juice and bile. The results point to the predominance of the secretory function of bile over pancreatic juice in sheep after stimulation of the duodenum with the investigated substances.

Animals

Bicarbonate and cyclic AMP content of pure human pancreatic juice in response to graded doses of synthetic secretin.

In seven healthy volunteers pure pancreatic juice was obtained by endoscopic cannulation of the papilla of Vater. Synthetic secretin was intravenously infused in doses doubled every 20 min. The volume of pancreatic juice was proportional to the log of the secretin dose. A significant rise (P less than 0.05) in pancreatic juice flow was elicited at a dose as low as 8.05 ng per kg per hr. Maximum flow rate approximating 250 mul per 5 min per kg of body weight was attained during infusion of 129 ng per kg per hr. At the same dose maximal bicarbonate outputs averaging 383 muEq per hr per kg of body weight were obtained, whereas bicarbonate ion concentration approached peak values (135 +/- 9 muEq per ml) at 32.2 ng per kg per hr. Bicarbonate concentrations showed a tendency to fall at supramaximal doses. The effect of increasing secretin doses on bicarbonate and cyclic AMP concentrations was remarkably similar (rs = 0.635, P less than 0.001) suggesting the participation of cyclic AMP in human pancreatic bicarbonate secretion.

Adult

Biochemical studies of pancreatic juice collected by duodenal aspiration and endoscopic cannulation of the main pancreatic duct.

Biochemical studies of pancreatic juice collected by duodenal aspiration and endoscopic cannulation of the main pancreatic duct were performed and compared in the same fasted normal subjects in seven cases. The comparison of the results shows that under the same stimulation (0.5 CU/kg/hr of GIH secretin and 75 ng/kg/hr of cerulein) the endoscopic cannulation of the main pancreatic duct provides the best values for the maximal secretory capacity. On the other hand, this technique is no better than our current method of duodenal aspiration for the exploration of the enzyme secretion.

Adult

Inhibition by somatostatin of secretin-stimulated pancreatic secretion in man: a study with pure pancreatic juice.

The action of somatostatin on compostition and flow rate of pure pancreatic juice obtained by endoscopic cannulation of the main pancreatic duct was evaluated in 5 healthy volunteers. Synthetic secretin (0.06 CU/kg-h) was intravenously infused throughout the 80-min study. Bicarbonate concentrations in pancreatic juice achieved constant levels (117 +/- 3 muEq/ml) after 10 min, whereas a steady state of juice flow (7.3 +/- 1.4 ml/5 min) was attained after 15 min of secretin infusion. In the third 20-min period, cyclic somatostatic (5 mug/kg-h i.v.) was given, leading to a decrease in pancreatic flow rate by 47% after 10 min, and by 67% after 15 min of somatostatin administration. Alrady 5 min after the infusion of somatostatin had been discontinued, pancreatic flow rate gradually recovered; presomatostatin levels, however, were not reached within 20 min. Cyclic AMP varied roughly in accordance with bicarbonate concentrations, whereas the chloride concentrations were reciprocally related. Bicarbonate, sodium, potassium, protein, and cyclic GMP concentrations did not change substantially due to somatostatin.

Adult

alpha-Amylase from human pancreatic juice as an electrophoretically pure isozyme.

Human pancreatic juice collected antiseptically and strictly free from other body fluids and juices, was examined for the isozyme composition of alpha-amylase. Unlike those of the extract of and exudate from pancreatic tissue, the composition of alpha-amylase isozyme in the secreted juice was simple and consisted of only two isozymes, a major one and a minor one, and the reason for difference in the isozyme composition is discussed. The major alpha-amylase isozyme was obtained in a crystalline state with a molecular weight of 5.4-5.5 X 10(4).

Amylases

The disintegration of surgical sutures on exposure to pancreatic juice.

The loop-breaking strength of various suture materials was tested over a period of 14 days during which time the sutures were incubated in vitro in saline or canine serum, bile, activated or nonactivated pancreatic juice. Under the conditions of the study, silk and nylon maintained their strength in each environment. Polyglycolic acid maintained its strength in saline, bile or serum, but gradually lost much of its strength when exposed to pancreatic juice. Catgut, both plain and chromic, disintegrated almost completely within 24-48 hours respectively when exposed to enterokinase activated pancreatic juice. Inhibition of trypsin by aprotinin (Trasylol) resulted in preservation of catgut strength but inhibition by soybean inhibitor did not. The latter findings suggest that proteolytic enzymes, other than trypsin, may be responsible for the disintegration.

Animals

Cyclic nucleotides, calcium, bicarbonate, and protein secretion in canine pancreatic juice in response to cholecystokinin-pancreozymin.

The secretion of cyclic AMP, cyclic GMP, protein, calcium, and bicarbonate in the pancreatic juice of three nonanesthetized dogs with chronic gastric and duodenal Thomas cannulae has been studied. Intravenous infusions of increasing doses of cholecystokinin-pancreozymin (CCK) (1.5, 3, 6, 12, 24 Crick Harper-Raper (CHR) U kg-1 h-1) were administered together with a continuous submaximal dose of secretin (1 clinical unit (CU) kg-1 h-1). Doubling CCK doses every 45 min induced a parallel increase in the output of both cyclic nucleotides. Cyclic AMP output peaked at between 15 and 30 min for 3 and 6 U kg-1 h-1 of CCK and later for 12 and 24 U kg-1 h-1 of CCK whereas cyclic GMP output increased more constantly. Calcium output followed a pattern similar to that of cyclic GMP secretion. Flow rate and protein output attained their peaks at between 30 and 45 min. A strong linear correlation was found between the quantities of cyclic AMP, cyclic GMP, and the quantities of protein secreted in response to each CCK dose. This study demonstrates the presence of cyclic GMP in the canine pancreatic juice and the dose-dependent stimulation of the secretion of cyclic GMP and cyclic AMP by CCK in the presence of secretin.

Animals

Lysosomal enzymes in pure pancreatic juice from normal healthy volunteers and chronic alcoholics.

Investigation of pure human pancreatic juice obtained by direct cannulation of the main pancreatic duct of 11 healthy volunteer subjects and 10 chronic alcoholics without detectable pancreatic disease revealed the presence of numerous acid hydrolases in this secretion. The pH optimal and substrate specificities of these enzymes suggest that they are of lysosomal origin. Stimulation of the pancreas by injection of cholecystokinin-pancreozymin (CCK-PZ) (1 Ivy dog unit/kg) resulted in a striking increase in activity of some of these hydrolases (N-acetyl-beta-D-glucosaminidase, arylsulfatase, etc.) similar to that observed for trypsin, amylase, and other pancreatic digestive enzymes. In a second group of hydrolases (beta-D-glucuronidase, leucine naphthylamidase, etc.) the effect of this hormone was greatly reduced or absent, particularly in normal individuals. In chronic alcoholics enzyme activity in response to CCK-PZ injection was greater than in normal subjects. Although this increase achieved statistical significance (P less than 0.05) in the case of beta-D-glucuronidase only, it was observed for all lysosomal hydrolases tested and suggests either increased synthesis or a more facile release of these enzymes from the pancreas of chronic alcoholics than of normal individuals.

Acetylglucosaminidase

Characterization and purification of a kallikrein from human pancreatic juice and immunological comparison with other kallikreins.

A prekallikrein has been demonstrated in human pancreatic juice and the active enzyme has been purified from this material. The purification procedure included filtration on Sephadex G-100, chromatography on DEAE-cellulose and affinity chromatography on trypsin-inhibitor Sepharose. The purified kallikrein appeared to be homogeneous by polyacrylamide gel electrophoresis at pH 8.3 and by immunoelectrophoresis. Human pancreatic kallikrein is immunologically different from human plasma kallikrein and from pancreatic kallikreins of other species (hog, cat, rat and dog). Human pancreatic kallikrein has common antigenic determinants with human urinary and submandibular kallikreins but probably not with parotid kallikrein.

Animals

[Effects of diets rich in triglycerides and phospholipids on lipase and phospholipase A activities in rat pancreatic juice and pancreas].

Lipase and phospholipase A activities were increased in the pancreas and pancreatic juice of rats ingesting during two months 20% phospholipids- or triglycerides-rich diets. The activity of lipase is higher with the triglycerides-rich diet and the phospholipase A activity is higher with the phospholipids-rich diet. After seven days of the experiment, these two enzymes activities were less increased with diet containing 40% lipids.

Animals