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Biomedical subjects

A Borgström

Publications and source records attributed to A Borgström.

At least 55 records · Page 3Linked to original sources

Protease activation following reperfusion of porcine pancreatic allografts.

To study the degree of protease activation at reperfusion of a pancreatic allograft after cold storage for 24 hr, 18 porcine whole-organ pancreaticoduodenal allograft transplantations were performed. Twelve grafts were flushed with and stored in Perfadex. In six of these, a hyperosmotic salt solution was injected into the graft aorta at reperfusion. Six grafts were flushed and stored in UW solution. Eleven of twelve grafts in the Perfadex groups were functioning on the first postoperative day, compared with one of six in the UW solution group. There was a significantly more pronounced protease activation among grafts stored in UW solution than in the other groups, with a subsequent breakthrough of the local protease protection barrier made up of protease inhibitors. In surviving pigs (n = 14), biochemical signs of protease activation evolved in plasma, including formation of trypsin-protease inhibitor complexes, a decline in C3 and kininogen levels, and a decline in functionally active alpha 2-macroglobulin, functionally active antithrombin III, and plasma kallikrein inhibitory activity. These biochemical signs of pancreatitis correlated with a deteriorated graft function on the second postoperative day, indicating that graft tissue damage occurred due to protease activation.

Animals↗

Characterization of immunoreactive trypsin as a means of differentiating graft pancreatitis and allograft rejection after porcine pancreatic transplantation.

Graft pancreatitis and allograft rejection were both accompanied by increased serum levels of immunoreactive anionic trypsin (irAT) in a porcine pancreatic allograft transplantation model. Characterization of this immunoreactivity by gel filtration revealed different elution profiles in these conditions that can be helpful in the differentiation between them. During graft pancreatitis, a major part of the immunoreactivity was found within the high-molecular-weight fraction corresponding to the formation of complexes between trypsin and protease inhibitors. During allograft rejection, virtually all serum irAT increase could be attributed to the release of anionic trypsinogen without any evidence of activation. Since this transplantation model includes urinary diversion of the exocrine secretions, irAT and immunoreactive cationic trypsin (irCT) can also be measured in the urine. Characterization of this immunoreactivity showed that most of both irAT and irCT was found as active trypsin but a minor part was probably complexed with some protease inhibitor (possibly pancreatic secretory trypsin inhibitor [PSTI]).

Animals↗

Duodenal mucosal pH as a reperfusion indicator in pancreatic-duodenal transplantation in the pig.

We have studied differences in reperfusion between the pancreas and the duodenum after 6 hr of cold storage in a porcine whole-organ pancreaticoduodenal allograft transplantation model. Two different flush-out and storage solutions, Perfadex and EuroCollins, were compared. Graft duodenal mucosal pH (pHi) was measured as an indicator of duodenal mucosal reperfusion. Pancreatic reperfusion was estimated indirectly using a ratio between the release of immunoreactive cationic trypsin (irCT) to serum 10 and 60 min following reperfusion. Twelve pigs (Perfadex n = 6, EuroCollins n = 6) were transplanted and all showed preserved endocrine and exocrine function postoperatively. Our data support the concept that reperfusion of a pancreaticoduodenal graft can be estimated using duodenal pHi and that the ratio of irCT at 10 and 60 min gives an indirect estimate of pancreatic reperfusion. The results also show that grafts stored in EuroCollins reperfuse more slowly than grafts stored in Perfadex.

Animals↗

Biochemical characterization of reperfusion pancreatitis in porcine pancreatic allografts after six hours of cold storage.

Biochemical signs of pancreatitis in plasma and pancreatic exudates were determined in 22 pigs subjected to pancreatic allograft transplantation after the graft had been in cold storage for 6 hr. Two perfusion and preservation media were used. We found signs of protease activation in the pancreatic exudate during the first hour after reperfusion. The local protease protection barrier was, however, not broken and no plasma changes indicating pancreatitis were seen during this period. On the first and second postoperative days, mild biochemical signs of pancreatitis were seen in the plasma, including a decrease in kininogen and C3 concentration as well as in plasma kallikrein inhibitory activity and the appearance of trypsin-protease inhibitor complexes. No correlation was seen between these biochemical signs of pancreatitis and graft appearance or function, indicating that the reperfusion pancreatitis seen after 6 hr of cold storage is of minor significance. No significant differences were seen between the two preservation media used (Perfadex and EuroCollins solution).

Animals↗

Early indicators of allograft rejection in a porcine pancreatic transplantation model.

Urine cytology and blood lymphocyte blastogenesis were evaluated as indicators of allograft rejection in a porcine pancreatic transplantation model. The percentage of activated lymphocytes and/or blasts was significantly increased during the rejection phase. Positive cytology was present in all rejection episodes. An increased thymidine uptake of blood lymphocytes and a decreased uridine/thymidine uptake quotient were seen prior to the onset of rejection. The reported dissociation of anionic and cationic trypsin levels in serum and urine after transplantation was not seen after simple urinary diversion of the pancreatic juice. This supports the hypothesis that a decreased synthesis of cationic trypsinogen compared with anionic trypsinogen occurs after porcine pancreatic transplantation.

Animals↗

Pancreatic cationic elastase in porcine experimental pancreatitis.

A radioimmunoassay for porcine, cationic, pancreatic elastase (irPE) is described. Normal porcine serum contains only small amounts of irPE (less than 3 micrograms/l). IrPE in serum and peritoneal exudate from 6 pigs with experimental pancreatitis was found mainly in a molecular form corresponding to free pro-enzyme. The presence of alpha 1-, alpha 2-macroglobulin-bound elastase-like enzymatic activity in the peritoneal exudates from pigs with pancreatitis, however, indicates that the proteinase is to some extent released as the active enzyme. In some pigs with pancreatitis, the elastase-like activity against Succ(Ala) in the peritoneal exudates increased during the experiment, arguing for a progressive activation of pro-elastase. Free proteolytic activity was not observed in any of the peritoneal exudates. This low degree of activation of elastase and the fact that the elastase inhibiting capacity is substantially larger than the trypsin inhibiting capacity in serum and biological fluids, leads us to the conclusion that active elastase is not a factor of principal importance in the pathogenesis of proteinase inhibitor consumption and tissue damage in our experimental pancreatitis model.

Animals↗

Porcine pancreatic cationic pro-elastase. Studies on the activation, turnover and interaction with plasma proteinase inhibitors.

Porcine pancreatic cationic pro-elastase was partly purified from pancreatic juice. The pro-enzyme binds slowly to alpha-macroglobulin and alpha 1-proteinase inhibitor. After 24 h incubation with plasma at room temperature more than 50% of the pro-elastase was still recovered in the form of free pro-enzyme. The pro-enzyme was activated by trypsin at neutral pH and by cathepsin B at pH 3.8. In the pig the half-life of i.v. administered pro-enzyme was about 30 min. After injection into the pancreatic duct radioactively labelled pro-elastase appeared in plasma within 30 min, and in peritoneal fluid after about 1 h.

Animals↗

Immunoreactive trypsin(ogen) in the sera of children with recent-onset insulin-dependent diabetes and matched controls. The Swedish Childhood Diabetes Group.

To evaluate the exocrine pancreatic function at the time of diagnosis of insulin-dependent diabetes mellitus, we determined immunoreactive anodal and cathodal trypsin(ogen) levels in sera from almost all children (n = 375) 0-14 years of age in Sweden in whom diabetes developed during 1 year, and in sex-, age-, and geographically matched control subjects (n = 312). The median level of anodal trypsin(ogen) was 5 (quartile range, 3-7) micrograms/L in children with newly diagnosed diabetes, compared with a median level of 7 (quartile range, 4-8) micrograms/L in control subjects (p less than 0.0001). Similarly, the median level of cathodal trypsin(ogen) was 8 (quartile range, 4-10) micrograms/L in children with diabetes, compared with a median level of 11 (quartile range, 7-15) micrograms/L in control subjects (p less than 0.0001). The median of the individual ratios between cathodal and anodal trypsin(ogen) was 1.4 in the diabetic patients and 1.7 in the control children (p less than 0.001). In a multivariate test, however, only the decrease in cathodal trypsin(ogen) concentration was associated with diabetes. The levels of trypsin(ogen)s did not correlate with levels of islet cell antibodies, present in 81% of the diabetic children. Several mechanisms may explain our findings, for example, similar pathogenetic factors may affect both the endocrine and exocrine pancreas simultaneously, a failing local trophic stimulation by insulin on the exocrine cells may decrease the trypsinogen production, and there may be an increased elimination of trypsin(ogen) because of higher filtration through the kidneys in the hyperglycemic state.

Adolescent↗

Serum immunoreactive anodal trypsinogen and urinary amylase as biochemical markers for rejection of clinical whole-organ pancreas allografts having exocrine drainage into the urinary bladder.

Rejection of pancreas allografts is best measured today by co-monitoring the creatinine of a simultaneously transplanted kidney allograft from the same donor. This methodology discourages pancreas transplantation for patients who have previously received a kidney allograft and preuremic patients. Thus, an early, graft-specific marker of rejection is desirable. In this study we compared 2 putative biochemical markers for rejection of pancreas allografts, serum immunoreactive anodal trypsinogen and urinary amylase, with serum creatinine in 15 simultaneously transplanted type I diabetics. Serial values during hospitalizations were determined. Follow-up ranged from 18 to 134 postoperative days. Rejection was diagnosed clinically and considered real if the patient received a course of anti-rejection medication. Ten of these 15 patients experienced a total of 21 rejection episodes. For all episodes of rejection, serum trypsinogen rose from a baseline of 398.1 +/- 25 ng/ml to 1686.2 +/- 317.9 ng/ml (P less than 0.001) on the day of rejection. Urinary amylase fell from 88,310 +/- 7877 U/24 hr to 37,508 +/- 7142 U/24 hr (P less than 0.001). For 10 patients in whom rejection was diagnosed on the initial hospitalization so that serial prediagnosis sera and urines were available, anodal trypsinogen rose from a baseline of 756 +/- 263 ng/ml to 1936 +/- 582 ng/ml (P less than 0.001). Urinary amylase values for these same 10 patients did not change significantly (baseline = 55,788 +/- 18,404 U/24 hr, rejection = 47,133 +/- 14,737 U/24 hr, (P = 0.7). We conclude that serum anodal trypsinogen behaves as a graft-specific biochemical marker for rejection of vascularized pancreas allografts.

Amylases↗

Urinary insulin level as an indicator of graft function after porcine pancreatic transplantation.

Urinary levels of immunoreactive insulin (IRI) were measured in 18 pigs subjected to pancreatic allograft transplantation with exocrine drainage into the urinary tract. Fifteen pigs were given no immuosuppressive therapy while 3 pigs received cyclosporine and prednisolone. The onset of rejection was defined as an increase in the serum levels of anionic trypsin (irAT). Urinary levels of IRI were compared between normo- and hyperglycemic pigs representing slow and fast rejectors. It was possible to measure insulin in the urine from all these pigs with a pancreatic allograft, and the urinary IRI levels increased after an intravenous injection of secretion and cholecystokinin. We found that urinary IRI response to secretin and cholecystokinin declined during rejection. By contrast, baseline, unstimulated urinary IRI levels did not correlate with rejection. No advantage was seen in the determination of urinary IRI when compared to determination of urinary irAT. In pigs not treated with immunosuppressants (with irreversible rejection), stimulated urinary levels of IRI and irAT were highly useful as graft-function indicators, whereas in immunosuppressed pigs (with reversible rejection episodes) they seemed to complement each other.

Animals↗

Indomethacin and pancreatic blood flow. An experimental study in pigs.

Indomethacin has been reported to decrease pancreatic blood flow. The drug has been used as an analgesic in acute pancreatitis. As decreased blood flow to the pancreas may detrimentally affect the outcome of acute pancreatitis, we investigated the effects of indomethacin on blood flow in the normal porcine pancreas. Regional blood flows, with special reference to the pancreatic flow, were studied with radioactively labelled microspheres in ketamine-anesthetized pigs before and after intravenous administration of indomethacin 2 mg/kg during 10 min. A transient decrease of cardiac output was seen during the infusion. Basal pancreatic blood flow was significantly increased 10 and 30 min after administration of indomethacin. No significant changes were found in small-intestinal or renal blood flow. We conclude that indomethacin does not reduce blood flow in normal porcine pancreas.

Animals↗

Immunoreactive anionic and cationic trypsin in human serum.

A simple method for the purification of anionic and cationic trypsinogen and trypsin from human pancreatic juice applying affinity chromatography on aprotinin coupled Sepharose is described together with the N-terminal amino acid sequences for both trypsinogens. In addition, enzyme-linked immunoabsorbent assay (ELISA) methods for the determination of anionic and cationic trypsin-like immunoreactivity (irAT and irCT) are described. Normal serum levels are 21.3 +/- 7.4 micrograms/l and 27.8 +/- 9.0 microgram/l for irAT and irCT respectively and the accuracy of these assays is 6-10%. In our population, the normal ratio between irCT and irAT in serum is 1.36 +/- 0.42. In normal serum trypsin-like immunoreactivity consists solely of trypsinogen. In acute pancreatitis there is an increase over normal of both irAT and irCT with a proportionally greater increase in irAT than irCT. Similar changes are also found in uremic patients.

Acute Disease↗

Severe acute pancreatitis and normal serum amylase activity due to pancreatic isoamylase deficiency.

A 31-year-old man with a primary attack of severe acute alcohol-induced pancreatitis presenting with a low to normal amylase activity in serum is described. The diagnosis was confirmed surgically and, further, by studies of immunoreactive trypsin in serum, which was elevated. Analysis of pancreatic isoamylase in serum during the convalescence showed very low activity. The patient is thought to represent a case of pancreatic isoamylase deficiency.

Acute Disease↗

Pancreatic enzymes in serum and urine as indicators of pancreatic allograft rejection in the pig.

We have studied the reliability of serum and urinary immunoreactive anionic trypsin (irAT), immunoreactive cationic trypsin (irCT), and amylase activity as rejection indicators in a porcine whole-organ pancreaticoduodenal transplantation model with exocrine drainage to the urinary bladder. No immunosuppressive therapy was administered. Exocrine tissue integrity and function were studied by measuring these enzymes in serum and urine. Urine analyses were performed before and after an intravenous secretin-cholecystokinin stimulation. Of 16 transplanted pigs, 10 became diabetic during a 2-week observation period while six remained normoglycemic. Serum irAT was found to predict rejection while serum amylase and serum irCT did not. An increase in irAT was seen in rejecting pigs preceding the onset of hyperglycemia by a median of 2 days (range 1-9). Secretion of irAT into the urine remained high during the observation period in nondiabetic pigs while the output declined in diabetic pigs. This decline was seen after the increase in serum irAT. When urine was sampled after a secretin-CCK stimulation, these findings were clearly evident, but less unequivocal results were obtained without stimulation. IrAT measurements were superior to measurements of amylase, irCT, or bicarbonate. Thus rejection of a pancreatic allograft was first indicated by a temporary rise in serum immunoreactive anionic trypsin, probably due to the onset of tissue damage. Thereafter, stimulated urinary enzyme output levels gradually declined and finally, hyperglycemia developed.

Amylases↗

Early diagnosis and classification in acute pancreatitis. A comparison of clinical outcome with findings at computed tomography and Ranson's prognostic signs.

The clinical outcome in 52 consecutive episodes of suspected acute pancreatitis was compared with Ranson's prognostic signs and findings on noncontrast and contrast-enhanced computed tomography (CT) scans performed within 24 h of patient admission. The predictive value of CT scan for diagnosis was 95% for positive results and 53% for negative results. In providing an accurate prognosis of a single attack, scoring of extrapancreatic signs was as good as that of Ranson's prognostic signs. CT with contrast medium revealed pancreatic ischaemia in 3 cases of 4 with clinically severe disease. The risk of developing severe pancreatitis was 23% in first attacks and 6% in relapses. Early CT scan is recommended in most patients with suspected pancreatitis to confirm diagnosis and to predict the severity of an attack.

Acute Disease↗

Pathobiochemistry and early CT findings in acute pancreatitis.

Extrapancreatic findings at computed tomography (CT), performed within 24 h in 42 consecutive episodes of acute pancreatitis, were classified according to a scoring system (EP score) and were correlated to Ranson's prognostic signs, to duration of hospital stay, biochemical changes in plasma and pancreatic ischaemia found at CT with contrast enhancement. Increasing EP score was found to be related to increasing number of positive Ranson's signs, longer hospital stay and pancreatic ischaemia. Plasma levels of immunoreactive cationic trypsin and amylase were not proportional to EP score. alpha 1-protease inhibitor, antichymotrypsin but not immunoreactive pancreatic secretory trypsin inhibitor increased proportionally to EP score. No changes related to EP score were seen in alpha 2-macroglobulin levels. Serum levels of trypsin-alpha 1-protease inhibitor complex were maximal after 3 days and most pronounced in cases with high EP scores. Plasma levels of factor X, alpha 2-antiplasmin and C1-esterase inhibitor were found to be inversely proportional to EP score.

Acute Disease↗

Markers for pancreas-graft rejection in humans.

Rejection episodes were studied in 15 patients, in whom no kidney graft could serve as a marker for rejection, subjected to pancreas transplantation with pancreatoenterostomy and temporary exteriorization of the pancreatic juice (10 pancreas alone, 3 pancreas after kidney, and 2 combined pancreas and kidney in which the kidney was not functioning.) Twelve patients (80%) had a total of 18 rejection episodes. In the first 11 patients, 13 rejection episodes were diagnosed by a decline in amylase activity in the pancreatic juice, whereas in the next 4 patients, 5 rejection episodes were diagnosed by positive cytology in the pancreatic juice. Neopterin in pancreatic juice and immunoreactive anionic trypsin in serum showed promise as rejection markers, whereas serum neopterin, serum amylase, and serum immunoreactive cationic trypsin did not. Unspecific signs of rejections were an increase in white blood cell count, clinical symptoms such as fever, abdominal pain, and arthralgia. All acute rejection episodes were successfully reversed by antirejection treatment. However, late rejections diagnosed by impaired endocrine function were seen in 6 of the 15 (40%) patients, and the prognoses for these rejections were worse: 4 patients (27%) lost their grafts because of chronic rejections, and 2 patients still had impaired endocrine function.

Amylases↗