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Biomedical subjects

G Appelt

Publications and source records attributed to G Appelt.

13 recordsLinked to original sources

Pancreatic duct mucosa following bile salt injury in cats. Morphology, barrier function to pancreatic exocrine proteins and vulnerability by activated pancreatic juice.

We studied pancreatic duct mucosal morphology and barrier function to both activated and nonactivated pancreatic exocrine proteins following bile salt injury in cats. Prograde pancreatic duct perfusion with 15 mM glycodeoxycholate caused epithelial disruption with focal epithelial loss in the majority of animals. In these cats, flow of preanalyzed nonactivated rat pancreatic juice along the duct resulted in a selective loss of zymogens from the duct lumen as determined by two-dimensional isoelectric focusing-sodium dodecyl sulfate gel electrophoresis and reversed-phase high-performance liquid chromatography. Proteins lost had a low isoelectric point (< or = 5.7) and a molecular weight as large as 47,000 Da and included chymotrypsinogen 1, trypsinogens 1 and 2, and procarboxypeptidases B. Proteins with a high isoelectric point (> 5.7), a high molecular weight (> 47,000 Da), or both, were completely recovered from the duct lumen. Flow of activated rat pancreatic juice along a pancreatic duct with bile salt-induced epithelial disruptions caused additional morphologic alterations including an increase in epithelial destruction and occasional necrosis of the duct interstitial and vascular tissue. In some cats, in which the integrity of the ductal epithelium remained preserved following exposure to the bile salt, neither loss of rat pancreatic exocrine zymogens from the duct lumen nor degradation of the duct mucosa by activated rat pancreatic juice was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Improved detection of human pancreatic proteins separated by two-dimensional isoelectric focusing/sodium dodecyl sulphate gel electrophoresis using a combined staining procedure.

In order to assess secretory pancreatic proteins in a two-dimensional isoelectric focusing/sodium dodecyl sulphate electrophoresis gel, a highly sensitive double-staining method with Coomassie Brilliant Blue followed by silver stain was used. This combined procedure afforded more distinct spots and additional bands, particularly glycoproteins, than either silver or Coomassie Blue staining alone. As measurements of dye volumes by densitometry have shown, double staining of two-dimensional separated pancreatic proteins is up to twenty times more sensitive than the usual Coomassie Brilliant Blue staining.

Electrophoresis, Gel, Two-Dimensional

[Isoelectric focusing of pure human pancreatic juice. 2. Identification of enzymes and proenzymes and characterization of glycoproteins].

After isoelectric focusing of proteins of human pure pancreatic juice following digestive enzymes and zymogens were identified by substrate specific detection methods: in the anodic range trypsinogen 1, and acidic form of proelastase and procarboxypeptidase A; in the pH--range of 5.0-6.0 a trypsinogen--like protein and the secretory trypsin inhibitor; in the pH--range of 6.0-7.0 one form of each of procarboxypeptidase B, trypsinogen 2, lipase, chymotrypsinogen and two forms of alpha-amylase and in the cathodic range an activated form of procarboxypeptidase B, one form of prophospholipase A2 and an additional form of proelastase. In order to characterize glycoproteins the IEF separated pancreatic proteins were transferred onto nitrocellulose membranes. Using the periodic acid/Schiff's reagent (PAS) procedure five PAS-positive proteins (pl s 4.5; 6.0; 6.6; 6.9 and 7.3) were detected. Three Con A-binding proteins (pl s 6.0; 6.6 and 6.9) were identified by incubating the blot in Con A followed by peroxidase and o-phenylendiamin as a chromogenic peroxidase substrat.

Enzyme Precursors

[Isoelectric focusing of pure human pancreatic juice. 1. Examination of patients with and without pancreatic diseases].

Thin layer urea isoelectric focusing is a powerful tool for a rapid and reproducible high resolution separation of cholecystokinin stimulated human pancreatic proteins. In pancreatic juice of patients without pancreatic disease, 11 main protein bands were separated and distinguished by Coomassie Blue-R 250 staining. A number of faint protein bands divided among the whole pH range demonstrate a wide microheterogeneity of exocrine pancreatic proteins. Secretory protein patterns of patients with chronic pancreatitis are shown to be different from those of patients without pancreatic disease. Significant alterations were found at the acidic protein band region of pI 4.4 to 4.7. In two cases of chronic pancreatitis the IEF pattern was very similar to that obtained by a short time enteropeptidase incubation of pure pancreatic juice from patients without pancreatic disease.

Cholecystokinin

Unexpected elution behaviour of peptides with various reversed-phase columns.

Gastrin and cholecystokinin peptides were separated by reversed-phase chromatography on conventional bristle-type and polymer-coated stationary phases. The retention of the sulphated and non-sulphated isomeric forms of both peptides is governed by the structure of the peptide, the net charge and additional polar interactions with the stationary phases. Polymer-coated phases are optimum for separations according to chain length, whereas polar interactions are required for the separation of sulphated and non-sulphated peptides of identical chain length.

Cholecystokinin

[Duodenal bile acid output in gastroenterologic diseases].

The biliary secretion of bile acids was measured with the modified secretin-pancreozymin test in 50 patients with various gastroenterological diseases and in a control group. In the control group the highest bile acid output and the lowest G/T quotient was estimated. In comparison to the control group, the bile acid output in the patients with gastroenterological disorders was reduced significantly to a varying degree. Except in postcholecystectomy patients, it was proved that the G/T quotient was significantly increased compared to that in the control group. The estimation of bile acid output and the G/T quotient presented here shows an easier extension of the validity of the secretin-pancreozymin test which does not indice any additional burden to the patients.

Adult

Analysis of human exocrine pancreatic proteins by means of pore gradient polyacrylamide gel electrophoresis.

Pancreatic juice collected by endoscopic retrograde cannulation of the pancreatic duct after administration of secretin and pancreozymin was analyzed by pore gradient polyacrylamide gel electrophoresis. Two electrophoretic systems, an alkaline (pH 8.9) and an acidic (pH 5.0), were elaborated to obtain optimal separation of exocrine proteins. 13 bands under alkaline conditions and 17 bands by running in the acidic system were separated and distinguished by Coomassie-Brilliant Blue G 250 staining. Fourteen proteins were identified using enzymatic and immunological methods: one form of each of lipase, chymotrypsinogen, prophospholipase A2, procarboxypeptidase A and trypsin inhibitor, two forms of each of procarboxypeptidase B and proelastase, three forms of each of trypsinogen and amylase. An additional band was identified as albumin.

Adult

[Pancreatico-biliary secretion in enteral hyperoxaluria].

The increased oxalic acid absorption is a well documented finding in gastroenterological diseases. The malabsorption of bile acids and fat is important in the pathogenesis of the hyperoxaluria. The enteric absorption of 14C-labelled oxalic acid was measured in 49 patients with different diseases. The dihydroxy-trihydroxy-ratio of bile acids is significantly decreased in patients with hyperoxaluria over 20% of the ingested dosage. We didn't find any correlation between the excretion of oxalic acid and the intraduodenal pancreatic lipase activity.

Adult

[Simultaneous registration of pancreatic secretion profiles in endoscopic secretion sampling in patients with and without pancreatic diseases].

A method for the continuous detection of secretory profiles of human pure pancreatic juice is described. After retrograde cannulation of the D. pancreaticus the exocrine pancreas was stimulated by an intravenous infusion of secretin and a bolus injection of pancreozymin additionally. The absorbance of pancreatic juice at 280 nm was measured continuously using an Uvicord-UV-Photometer which was connected directly with a Teflon catheter localised in the main pancreatic duct. The secretory profiles obtained from patients without pancreatic diseases (n = 12) indicated a first maximum of absorbance after secretin infusion and a second one in response to pancreozymin bolus injection. Alterations of secretory profiles in patients with chronic pancreatitis (n = 10) were evident after pancreozymin administration principally.

Adult

[Secretin-pancreozymin test with volume correction in the functional diagnosis of pancreatobiliary secretion].

A modified secretin-pancreozymin test was performed for the quantitative determination of both the pancreatic exocrine function and the biliary secretion of total bile acids followed by exogenous (secretin and pancreozymin) and endogenous (L-phenylalanine) stimulation of the pancreatic and biliary secretion. The outputs of lipase, alpha-amylase, trypsin, bicarbonate and total bile acids were estimated and the ratio of glycine to taurine conjugated bile acids was measured by thin layer chromatography. Polyethylene glycol 4,000 was used as a nonabsorbable marker for the correction of aspirated volumes. 40 patients were studied: 10 control subjects; 10 control subjects with previous cholecystectomy; 10 patients with chronic pancreatitis and 10 cholecystectomized patients with chronic pancreatitis. In cholecystectomized probands, not only the total bile acid output but also the output of bicarbonate, trypsin and lipase were diminished compared to the control subjects with a intact gallbladder. In noncholecystectomized and cholecystectomized patients with chronic pancreatitis both, the pancreatic secretion and the biliary bile acid secretion were significantly decreased. The ratio of glycine/taurine conjugated bile acids was found to be significantly higher in these patients compared to the controls.

Adult

Partition of purified human thyroxine-binding globulin in aqueous two-phase systems in response to reactive dyes.

Thyroxine-binding globulin was isolated from pooled human serum by a two-step method involving affinity chromatography on thyroxine-Sepharose and preparative disc-electrophoresis. The final product was found to be homogeneous by polyacrylamide gel electrophoresis and has a molecular weight of 59,000. Isoelectric focusing resolved the protein into seven bands with isoelectric points ranging from 3.9 to 4.3. The isolated protein showed affinity to a number of different dyes as recognized by affinity phase partitioning. The interaction of the protein with the dye Cibacron Blue F3G-A was found to be strongly competitive with the natural ligand thyroxine.

Albumins

Metastases from testicular carcinoma. Study of 78 autopsied cases.

The necropsy records of 78 patients with histologically proved germ cell tumors of the testis, who died as a direct result of their malignant disease, were reviewed to determine the usual modes of spread, distribution of metastasis, the histologic characteristics of the metastatic foci as compared with the morphology of the primary tumor and the specific cause of death. The sites of metastases in order of decreasing frequency for all cases were lung, retroperitoneal lymph nodes, liver, mediastinal lymph nodes, brain, kidney, gastrointestinal tract, bones, adrenals, peritoneum and spleen. The absence of metastases solely in the anterior mediastinum without involvement of other mediastinal nodes (middle/posterior) strongly supports the premise for a primary extragonadal origin whenever the anterior mediastinum alone is involved with malignant disease having the histologic appearance of a primary germ cell tumor. The histologic features of the metastatic lesions were usually similar in nature to those of the primary tumor except for seminoma in which the metastatic lesions proved to be of a different histologic pattern in almost one third of the patients dying from the disease. It should be axiomatic that whenever a patient with seminoma fails to respond appropriately to radiotherapy that his treatment be immediately discontinued and that appropriate biopsies be obtained to substantiate the histologic pattern present.

Adolescent