trypsinogen and Pancreatitis

Acute pancreatitis (AP) is a complex disease and the leading cause of gastrointestinal disease-related hospital admissions. Few therapeutic options exist for AP prevention. Blood proteins with causal evidence may represent promising drug targets, but few have been causally linked with AP. Our objective was to identify blood proteins linked with AP by combining genome-wide association meta-analysis and proteome-wide Mendelian randomization (MR) studies.. We performed a genome-wide association meta-analysis totalling 10,630 patients with AP and 844,679 controls and a series of inverse-variance weighted MR analyses using cis-acting variants on 4719 blood proteins from the deCODE study (N = 35,559) and 4979 blood proteins from the Fenland study (N = 10,708).. This large genome-wide association study meta-analysis for AP identified new variants linked with AP as well as several blood proteins that may be causally associated with AP. This study provides new information on the genetic architecture of this disease and identified pathways related to AP, which may be further explored as possible therapeutic targets for AP.

Topics: Acute Disease; Blood Proteins; Genome-Wide Association Study; Humans; Mendelian Randomization Analysis; Nuclear Proteins; Pancreatitis; Polymorphism, Single Nucleotide; Proteome; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Not all patients with severe hypertriglyceridemia develop acute pancreatitis. We surveyed recent literature on inter-individual genetic variation in susceptibility to pancreatitis.. Genetic determinants of pancreatitis include: rare Mendelian disorders caused by highly penetrant pathogenic variants in genes involved in trypsinogen activation; uncommon susceptibility variants in genes involved in trypsinogen activation, protein misfolding as well as calcium metabolism and cystic fibrosis, that have variable penetrance and show a range of odds ratios for pancreatitis; and common polymorphisms in many of the same genes that have only a small effect on risk. The role of these genetic variants in modulating pancreatitis risk in hypertriglyceridemia is unclear. However, among genetic determinants of plasma triglycerides, those predisposing to more severe hypertriglyceridemia associated with chylomicronemia appear to have higher pancreatitis risk.. Currently, among patients with severe hypertriglyceridemia, the most consistent predictor of pancreatitis risk is the triglyceride level. Furthermore, pancreatitis risk appears to be modulated by a higher genetic burden of factors associated with greater magnitude of triglyceride elevation. The role of common and rare genetic determinants of pancreatitis itself in this metabolic context is unclear.

Topics: Acute Disease; Humans; Hypertriglyceridemia; Pancreatitis; Triglycerides; Trypsinogen

Trypsinogen (PRSS1, PRSS2) copy number gains and regulatory variants have both been proposed to elevate pancreatitis risk through a gene dosage effect (i.e., by increasing the expression of wild-type protein). However, to date, their impact on pancreatitis risk has not been thoroughly evaluated whilst the underlying pathogenic mechanisms remain to be explicitly investigated in mouse models. Genetic studies of the rare trypsinogen duplication and triplication copy number variants (CNVs), and the common rs10273639C variant, were collated from PubMed and/or ClinVar. Mouse studies that analyzed the influence of a transgenically expressed wild-type human PRSS1 or PRSS2 gene on the development of pancreatitis were identified from PubMed. The genetic effects of the different risk genotypes, in terms of odds ratios, were calculated wherever appropriate. The genetic effects of the rare trypsinogen duplication and triplication CNVs were also evaluated by reference to their associated disease subtypes. We demonstrate a positive correlation between increased trypsinogen gene dosage and pancreatitis risk in the context of the rare duplication and triplication CNVs, and between the level of trypsinogen expression and disease risk in the context of the heterozygous and homozygous rs10273639C-tagged genotypes. We retrospectively identify three mouse transgenic studies that are informative in relation to the pathogenic mechanism underlying the trypsinogen gene dosage effect in pancreatitis. Trypsinogen gene dosage correlates with pancreatitis risk across genetic and transgenic studies, highlighting the fundamental role of dysregulated expression of wild-type trypsinogen in the etiology of pancreatitis. Specifically downregulating trypsinogen expression in the pancreas may serve as a potential therapeutic and/or prevention strategy for pancreatitis.

Topics: Animals; Animals, Genetically Modified; Gene Dosage; Humans; Mice; Mutation; Pancreatitis; Retrospective Studies; Trypsin; Trypsinogen

Recent studies have highlighted the importance of autophagy and particularly non-canonical autophagy in the development and progression of acute pancreatitis (a frequent disease with considerable morbidity and significant mortality). An important early event in the development of acute pancreatitis is the intrapancreatic activation of trypsinogen, (i.e., formation of trypsin) leading to the autodigestion of the organ. Another prominent phenomenon associated with the initiation of this disease is vacuolisation and specifically the formation of giant endocytic vacuoles in pancreatic acinar cells. These organelles develop in acinar cells exposed to several inducers of acute pancreatitis (including taurolithocholic acid and high concentrations of secretagogues cholecystokinin and acetylcholine). Notably, early trypsinogen activation occurs in the endocytic vacuoles. These trypsinogen-activating organelles undergo activation, long-distance trafficking, and non-canonical autophagy. In this review, we will discuss the role of autophagy in acute pancreatitis and particularly focus on the recently discovered LAP-like non-canonical autophagy (LNCA) of endocytic vacuoles.

Topics: Acute Disease; Autophagy; Humans; Pancreatitis; Trypsinogen; Vacuoles

The focus of the review is on roles of autophagy and pancreatic secretory trypsin inhibitor (PSTI), an endogenous trypsin inhibitor, in trypsinogen activation in acute pancreatitis. Acute pancreatitis is a disease in which tissues in and around the pancreas are autodigested by pancreatic digestive enzymes. This reaction is triggered by the intrapancreatic activation of trypsinogen. Autophagy causes trypsinogen and cathepsin B, a trypsinogen activator, to colocalize within the autolysosomes. Consequently, if the resultant trypsin activity exceeds the inhibitory activity of PSTI, the pancreatic digestive enzymes are activated, and they cause autodigestion of the acinar cells. Thus, autophagy and PSTI play important roles in the development and suppression of acute pancreatitis, respectively.

Topics: Acinar Cells; Animals; Autophagy; Cathepsin B; Disease Models, Animal; Endoplasmic Reticulum Stress; Enzyme Activation; Glycoproteins; Humans; Lysosomes; Mice; Mice, Knockout; Molecular Chaperones; Pancreatitis; Prostatic Secretory Proteins; Protein Folding; Proteolysis; Secretory Vesicles; Transcription Factor CHOP; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Genetic association studies regarding relationship between PRSS1-PRSS2 rs10273639/CLDN2 rs7057398/MORC4 rs12688220 polymorphisms and pancreatitis yielded conflicting results. We performed this meta-analysis to explore associations between these polymorphisms and pancreatitis in a larger pooled population.. A systematic search of the literature was conducted for eligible studies. We used Review Manager to conduct statistical analyses.. Fifteen studies were included in this meta-analysis. The results of pooled analyses showed that CLDN2 rs7057398, MORC4 rs12688220 and PRSS1-PRSS2 rs10273639 polymorphisms were all significantly associated with susceptibility to acute pancreatitis in Caucasians. Moreover, MORC4 rs12688220 and PRSS1-PRSS2 rs10273639 polymorphisms were also significantly associated with susceptibility to chronic pancreatitis in Asians.. Our findings suggested that rs7057398, rs12688220 and rs10273639 polymorphisms could be used to identify individuals at an elevated susceptibility to acute pancreatitis in Caucasians. Moreover, rs12688220 and rs10273639 polymorphisms could be used to identify individuals at an elevated susceptibility chronic pancreatitis in Asians.

Topics: Asian People; Claudins; Genetic Association Studies; Genetic Predisposition to Disease; Humans; Nuclear Proteins; Pancreatitis; Pancreatitis, Chronic; Polymorphism, Genetic; Trypsin; Trypsinogen; White People

Premature activation of digestive enzymes in the pancreas has been linked to development of pancreatitis for more than a century. Recent development of novel models to study the role of pathologic enzyme activation has led to advances in our understanding of the mechanisms of pancreatic injury. Colocalization of zymogen and lysosomal fraction occurs early after pancreatitis-causing stimulus. Cathepsin B activates trypsinogen in these colocalized organelles. Active trypsin increases permeability of these organelles resulting in leakage of cathepsin B into the cytosol leading to acinar cell death. Although trypsin-mediated cell death leads to pancreatic injury in early stages of pancreatitis, multiple parallel mechanisms, including activation of inflammatory cascades, endoplasmic reticulum stress, autophagy, and mitochondrial dysfunction in the acinar cells are now recognized to be important in driving the profound systemic inflammatory response and extensive pancreatic injury seen in acute pancreatitis. Chymotrypsin, another acinar protease, has recently been shown be play critical role in clearance of pathologically activated trypsin protecting against pancreatic injury. Mutations in trypsin and other genes thought to be associated with pathologic enzyme activation (such as serine protease inhibitor 1) have been found in familial forms of pancreatitis. Sustained intra-acinar activation of nuclear factor κB pathway seems to be key pathogenic mechanism in chronic pancreatitis. Better understanding of these mechanisms will hopefully allow us to improve treatment strategies in acute and chronic pancreatitis.

Topics: Acinar Cells; Animals; Cell Death; Enzyme Activation; Genetic Predisposition to Disease; Humans; Inflammation Mediators; Mutation; Pancreas, Exocrine; Pancreatitis; Phenotype; Signal Transduction; Trypsin; Trypsinogen

The incidence of acute pancreatitis continues to increase worldwide, and it is one of the most common gastrointestinal causes for hospital admission in the USA. In the past decade, substantial advancements have been made in our understanding of the pathophysiological mechanisms of acute pancreatitis. Studies have elucidated mechanisms of calcium-mediated acinar cell injury and death and the importance of store-operated calcium entry channels and mitochondrial permeability transition pores. The cytoprotective role of the unfolded protein response and autophagy in preventing sustained endoplasmic reticulum stress, apoptosis and necrosis has also been characterized, as has the central role of unsaturated fatty acids in causing pancreatic organ failure. Characterization of these pathways has led to the identification of potential molecular targets for future therapeutic trials. At the patient level, two classification systems have been developed to classify the severity of acute pancreatitis into prognostically meaningful groups, and several landmark clinical trials have informed management strategies in areas of nutritional support and interventions for infected pancreatic necrosis that have resulted in important changes to acute pancreatitis management paradigms. In this Review, we provide a summary of recent advances in acute pancreatitis with a special emphasis on pathophysiological mechanisms and clinical management of the disorder.

Topics: Acute Disease; Animals; Calcium Signaling; Disease Management; Disease Models, Animal; Endoplasmic Reticulum Stress; Humans; Mutation; Nutritional Support; Pancreatitis; Severity of Illness Index; Terminology as Topic; Trypsinogen

The treatment of people with acute abdominal pain differs if they have acute pancreatitis. It is important to know the diagnostic accuracy of serum amylase, serum lipase, urinary trypsinogen-2, and urinary amylase for the diagnosis of acute pancreatitis, so that an informed decision can be made as to whether the person with abdominal pain has acute pancreatitis. There is currently no Cochrane review of the diagnostic test accuracy of serum amylase, serum lipase, urinary trypsinogen-2, and urinary amylase for the diagnosis of acute pancreatitis.. To compare the diagnostic accuracy of serum amylase, serum lipase, urinary trypsinogen-2, and urinary amylase, either alone or in combination, in the diagnosis of acute pancreatitis in people with acute onset of a persistent, severe epigastric pain or diffuse abdominal pain.. We searched MEDLINE, Embase, Science Citation Index Expanded, National Institute for Health Research (NIHR HTA and DARE), and other databases until March 2017. We searched the references of the included studies to identify additional studies. We did not restrict studies based on language or publication status, or whether data were collected prospectively or retrospectively. We also performed a 'related search' and 'citing reference' search in MEDLINE and Embase.. We included all studies that evaluated the diagnostic test accuracy of serum amylase, serum lipase, urinary trypsinogen-2, and urinary amylase for the diagnosis of acute pancreatitis. We excluded case-control studies because these studies are prone to bias. We accepted any of the following reference standards: biopsy, consensus conference definition, radiological features of acute pancreatitis, diagnosis of acute pancreatitis during laparotomy or autopsy, and organ failure. At least two review authors independently searched and screened the references located by the search to identify relevant studies.. Two review authors independently extracted data from the included studies. The thresholds used for the diagnosis of acute pancreatitis varied in the trials, resulting in sparse data for each index test. Because of sparse data, we used -2 log likelihood values to determine which model to use for meta-analysis. We calculated and reported the sensitivity, specificity, post-test probability of a positive and negative index test along with 95% confidence interval (CI) for each cutoff, but have reported only the results of the recommended cutoff of three times normal for serum amylase and serum lipase, and the manufacturer-recommended cutoff of 50 mg/mL for urinary trypsinogen-2 in the abstract.. Ten studies including 5056 participants met the inclusion criteria for this review and assessed the diagnostic accuracy of the index tests in people presenting to the emergency department with acute abdominal pain. The risk of bias was unclear or high for all of the included studies. The study that contributed approximately two-thirds of the participants included in this review was excluded from the results of the analysis presented below due to major concerns about the participants included in the study. We have presented only the results where at least two studies were included in the analysis.Serum amylase, serum lipase, and urinary trypsinogen-2 at the standard threshold levels of more than three times normal for serum amylase and serum lipase, and a threshold of 50 ng/mL for urinary trypsinogen-2 appear to have similar sensitivities (0.72 (95% CI 0.59 to 0.82); 0.79 (95% CI 0.54 to 0.92); and 0.72 (95% CI 0.56 to 0.84), respectively) and specificities (0.93 (95% CI 0.66 to 0.99); 0.89 (95% CI 0.46 to 0.99); and 0.90 (95% CI 0.85 to 0.93), respectively). At the median prevalence of 22.6% of acute pancreatitis in the studies, out of 100 people with positive test, serum amylase (more than three times normal), serum lipase (more than three times normal), and urinary trypsinogen (more than 50 ng/mL), 74 (95% CI 33 to 94); 68 (95% CI 21 to 94); and 67 (95% CI 57 to 76) people have acute pancreatitis, respectively; out of 100 people with negative test, serum amylase (more than three times normal), serum lipase (more than three times normal), and urinary trypsinogen (more than 50 ng/mL), 8 (95% CI 5 to 12); 7 (95% CI 3 to 15); and 8 (95% CI 5 to 13) people have acute pancreatitis, respectively. We were not able to compare these tests formally because of sparse data.. As about a quarter of people with acute pancreatitis fail to be diagnosed as having acute pancreatitis with the evaluated tests, one should have a low threshold to admit the patient and treat them for acute pancreatitis if the symptoms are suggestive of acute pancreatitis, even if these tests are normal. About 1 in 10 patients without acute pancreatitis may be wrongly diagnosed as having acute pancreatitis with these tests, therefore it is important to consider other conditions that require urgent surgical intervention, such as perforated viscus, even if these tests are abnormal.The diagnostic performance of these tests decreases even further with the progression of time, and one should have an even lower threshold to perform additional investigations if the symptoms are suggestive of acute pancreatitis.

Topics: Acute Disease; Amylases; Biomarkers; Diagnostic Errors; Humans; Lipase; Pancreatitis; Trypsin; Trypsinogen

Type 1 diabetes (T1D) is considered a pancreatic beta cell-specific disease that results in absolute insulin deficiency. Nevertheless, clinical studies from 1940 onwards showed that patients with T1D had an abnormal exocrine pancreas due to the presence of subclinical exocrine insufficiency and acinar atrophy. Exocrine abnormalities are an important, and mostly neglected, characteristic associated with T1D. It is however still unclear whether the exocrine dysfunction in T1D is a primary damage caused by the same pathogenic event that led to beta cell destruction or secondary to beta cell loss. In this review, we collect evidence supporting the hypothesis that T1D is a combined endocrine-exocrine disease in which the loss of functional beta cell mass is most clinically apparent.

Topics: Animals; Diabetes Mellitus, Type 1; Humans; Insulin-Secreting Cells; Pancreas, Exocrine; Pancreatic Elastase; Pancreatitis; Trypsinogen

Chronic pancreatitis is a progressive inflammatory disease in which pancreatic secretory parenchyma is destroyed and replaced by fibrous tissue, eventually leading to malnutrition and diabetes. Alcohol is the leading cause in Western countries, but genetic factors are also implicated. Since the identification of mutations in the cationic trypsinogen (PRSS1) gene as a cause of hereditary pancreatitis in 1996, we have seen great progress in our understanding of the genetics of pancreatitis. It has been established that mutations in the genes related to the activation and inactivation of trypsin(ogen) such as PRSS1, serine protease inhibitor Kazal type 1 (SPINK1) and chymotrypsin C (CTRC) genes are associated with pancreatitis. In 2013, carboxypeptidase A1 (CPA1) was identified as a novel pancreatitis susceptibility gene. Endoplasmic reticulum stress in pancreatic acinar cells resulting from the mis-folding of mutated pancreatic enzymes has been shown to act as a novel mechanism underlying the susceptibility to pancreatitis. In Japan, the nationwide survey revealed 171 patients (96 males and 75 females) with hereditary pancreatitis in 59 families based on the European Registry of Hereditary Pancreatitis and Familial Pancreatic Cancer criteria. Because about 30% of families with hereditary pancreatitis do not carry mutations in any of the known pancreatitis susceptibility genes, other yet unidentified genes might be involved. Next generation sequencers can perform billions of sequencing reactions with a read length of 150-250 nucleotides. Comprehensive analysis using next generation sequencers will be a promising strategy to identify novel pancreatitis-associated genes and further clarify the pathogenesis of pancreatitis.

Topics: Carboxypeptidases A; Carrier Proteins; Chymotrypsin; Endoplasmic Reticulum Stress; Exome; Female; Genetic Predisposition to Disease; Genotype; High-Throughput Nucleotide Sequencing; Humans; Japan; Male; Mutation; Pancreatitis; Surveys and Questionnaires; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

In this article, we review important advances in our understanding of the mechanisms of pancreatitis.. The relative contributions of intrapancreatic trypsinogen activation and nuclear factor kappa B (NFκB) activation, the two major early independent cellular events in pancreatitis, have been investigated using novel genetic models. Trypsinogen activation has traditionally held the spotlight for many decades as the central pathogenic event of pancreatitis. However, recent experimental evidence points to the role of trypsin activation in early acinar cell damage but not in the inflammatory response of acute pancreatitis, which was shown to be induced by NFκB activation. Further, chronic pancreatitis developed independently of trypsinogen activation in the caerulein model. Sustained NFκB activation, but not persistent intra-acinar expression of active trypsin, was shown to result in chronic pancreatitis. Calcineurin-NFAT (nuclear factor of activated T-cells) signaling was shown to mediate downstream effects of pathologic rise in intracellular calcium. Interleukin-6 was identified as a key cytokine mediating pancreatitis-associated lung injury.. Recent advances challenge the long-believed trypsin-centered understanding of pancreatitis. It is becoming increasingly clear that activation of intense inflammatory signaling mechanisms in acinar cells is crucial to the pathogenesis of pancreatitis, which may explain the strong systemic inflammatory response in pancreatitis.

Topics: Acute Disease; Genetic Predisposition to Disease; Humans; Mutation; NF-kappa B; Pancreatitis; Pancreatitis, Chronic; Signal Transduction; Systemic Inflammatory Response Syndrome; Trypsin; Trypsinogen

Currently, serum amylase and lipase are the most popular laboratory markers for early diagnosis of acute pancreatitis with reasonable sensitivity and specificity. Urinary trypsinogen-2 (UT-2) has been increasingly used but its clinical value for the diagnosis of acute pancreatitis and post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis has not yet been systematically assessed.. A comprehensive search was carried out using PubMed (MEDLINE), Embase, and Web of Science for clinical trials, which studied the usefulness of UT-2 as a diagnostic marker for acute pancreatitis. Sensitivity, specificity and the diagnostic odds ratios (DORs) with 95% confidence interval (CI) were calculated for each study and were compared with serum amylase and lipase. Summary receiver-operating curves were conducted and the area under the curve (AUC) was evaluated.. A total of 18 studies were included. The pooled sensitivity and specificity of UT-2 for the diagnosis of acute pancreatitis were 80% and 92%, respectively (AUC=0.96, DOR=65.63, 95% CI: 31.65-139.09). The diagnostic value of UT-2 was comparable to serum amylase but was weaker than serum lipase. The pooled sensitivity and specificity for the diagnosis of post-ERCP pancreatitis were 86% and 94%, respectively (AUC=0.92, DOR=77.68, 95% CI: 24.99-241.48).. UT-2 as a rapid test could be potentially used for the diagnosis of post-ERCP pancreatitis and to an extent, acute pancreatitis. Further studies are warranted to confirm these results.

Topics: Amylases; Biomarkers; Cholangiopancreatography, Endoscopic Retrograde; Humans; Lipase; Pancreatitis; Sensitivity and Specificity; Trypsin; Trypsinogen

Urinary trypsinogen-2 has been implicated as a promising biomarker for the early diagnosis of acute pancreatitis (AP). The meta-analysis was used to establish the overall accuracy of urinary trypsinogen-2 test for diagnosing AP.. Based on comprehensive searches of the PubMed and Embase databases, we identified and abstracted outcome data from all articles evaluating the diagnostic value of urinary trypsinogen-2. A summary estimate for sensitivity, specificity, 95% confidence region and 95% prediction region was calculated using the bivariate random-effects approach.. The meta-analysis included 13 studies (2342 patients, the proportion of severe AP from 13.21% to 30.00%). Overall, the pooled sensitivity was 82.3% (95%CI 79.3%-85.1%) and specificity was 93.5% (95%CI 92.2%-94.6%). The diagnostic odds ratios (DOR) was 85.23 (95%CI 40.14-180.99). The area under the summary ROC curve (AUC) was 0.9673.. The urinary trypsinogen-2 test is a reliable and rapid method for the early diagnosis of AP.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Area Under Curve; Biomarkers; Confidence Intervals; Databases, Factual; Early Diagnosis; Female; Humans; Male; Middle Aged; Odds Ratio; Pancreatitis; Publication Bias; Reagent Kits, Diagnostic; Reproducibility of Results; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen; Young Adult

In this article, recent advances in the pathogenesis of acute pancreatitis have been reviewed.. Pathologic intra-acinar trypsinogen activation had been hypothesized to be the central mechanism of pancreatitis for over a century. This hypothesis could be explored for the first time with the development of a novel mouse model lacking pathologic intra-acinar trypsinogen activation. It became clear that intra-acinar trypsinogen activation contributes to early acinar injury, but local and systemic inflammation progress independently during pancreatitis. Early intra-acinar nuclear factor kappa B (NFκB) activation, which occurs parallel to but independent of trypsinogen activation, may be crucial in pancreatitis. Although the mechanism of NFκB and trypsinogen activation is not entirely clear, further insights have been made into key pathogenic cellular events such as calcium signaling, mitochondrial dysfunction, endoplasmic reticulum (ER) stress, autophagy and impaired trafficking, and lysosomal and secretory responses. Cellular intrinsic damage-sensing mechanisms that lead to activation of the inflammatory response aimed at repair, but lead to disease when overwhelmed, are beginning to be understood.. New findings necessitate a paradigm shift in our understanding of acute pancreatitis. Intra-acinar trypsinogen activation leads to early pancreatic injury, but the inflammatory response of acute pancreatitis develops independently, driven by early activation of inflammatory pathways.

Topics: Acinar Cells; Animals; Autophagy; Calcium; Humans; Inflammasomes; Mitochondria; NF-kappa B; Pancreatitis; Trypsinogen

Acute pancreatitis (AP) is an important cause of morbidity and mortality worldwide and the annual incidence appears to be increasing. It presents as a mild self-limiting illness in 80% of patients. However, one-fifth of these develop a severe complicated life-threatening disease requiring intensive and prolonged therapeutic intervention. Alcohol and gallstone disease remain the commonest causes of AP but metabolic abnormalities, obesity and genetic susceptibility are thought be increasingly important aetiological factors. The prompt diagnosis of AP and stratification of disease severity is essential in directing rapid delivery of appropriate therapeutic measures. In this review, the range of diagnostic and prognostic assays, severity scoring systems and radiological investigations used in current clinical practice are described, highlighting their strengths and weaknesses. Increased understanding of the complex pathophysiology of AP has generated an array of new potential diagnostic assays and these are discussed. The multidisciplinary approach to management of severe pancreatitis is outlined, including areas of controversy and novel treatments.

Topics: Acute Disease; Alcohol Drinking; Amylases; Autolysis; Gallstones; Genetic Predisposition to Disease; Humans; Hypercalcemia; Hyperlipidemias; Lipase; Obesity; Pancreatitis; Prognosis; Trypsin; Trypsinogen

IgG4-related systemic disease (ISD) is a recently recognized syndrome affecting multiple organs. Autoimmune pancreatitis (AIP) is the pancreatic manifestation of ISD and mimics pancreatic cancer. Current data show frequent association with serum IgG4 elevation and other serologic abnormalities. Here we explore the diagnostic and possible prognostic utility and pathogenetic implications of serologic abnormalities in ISD.. Serum IgG4 elevations (>140 mg/dl) are seen in 70-80% of AIP patients and also in 5% of normal population and 10% of pancreatic cancer making it an unsuitable single marker for diagnosis. However, when combined with other features of AIP, it can be of great diagnostic value though its utility in monitoring of therapy or as a marker or predictor of relapse is limited. Several other antibodies have been identified in AIP against pancreas-specific antigens like trypsinogens I and II, pancreatic secretory trypsin inhibitor (PSTI) and plasminogen binding protein (PBP) and other nonpancreas-specific antigens. Anti-PBP antibodies appear to have potential diagnostic utility but require further validation.. No single serologic marker is diagnostic of ISD. Serum IgG4 elevation has convincing diagnostic utility when combined with other disease features although its value in disease monitoring may be limited.

Topics: Antibody Specificity; Autoantibodies; Autoantigens; Autoimmune Diseases; Biomarkers; Carrier Proteins; Humans; Immunoglobulin G; Pancreas; Pancreatitis; Plasminogen; Trypsinogen

Topics: Acute Disease; Alleles; Carrier Proteins; Cytokines; Humans; Immune System Diseases; Inflammation Mediators; Lipopolysaccharide Receptors; Macrophage Migration-Inhibitory Factors; Mutation; Obesity; Pancreatitis; Severity of Illness Index; Toll-Like Receptors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Acute pancreatitis is an inflammatory disease of the pancreas which, in its most severe form, is associated with multi-organ failure and death. Recently, signalling molecules and pathways which are responsible for the initiation and progression of this disease have been under intense scrutiny. One important signalling molecule, nuclear factor kappaB (NF-kappaB), has been shown to play a critical role in the development of acute pancreatitis. NF-kappaB is a nuclear transcription factor responsible for regulating the transcription of a wide variety of genes involved in immunity and inflammation. Many of these genes have been implicated as central players in the development and progression of acute pancreatitis. This review discusses recent advances in the investigation of pancreatic and extrapancreatic (lungs, liver, monocytes and macrophages, and endothelial cells) NF-kappaB activation as it relates to acute pancreatitis.

Topics: Acute Disease; Arginine; Cell Communication; Cholecystokinin; Endothelial Cells; Humans; Ligation; Liver; Lung; Lymphocyte Activation; Macrophage Activation; Macrophages; Monocytes; NF-kappa B; Pancreatitis; Taurocholic Acid; Transcription Factor RelA; Trypsinogen

There was remarkable progress in the understanding of the role genetic risk factors in chronic pancreatitis. These factors seem to be much more important than thought in the past. The rare autosomal-dominant mutations N29I and R122H of PRSS1 (cationic trypsinogen) as well as the variant N34S of SPINK1 (pancreatic secretory trypsin inhibitor) are associated to a disease onset in childhood or youth. Compared to chronic alcoholic pancreatitis the progression is slow so that for a long time only signs of acute-recurrent pancreatitis are found. Only at later time points (more than 10-15 years) there is evidence for chronic pancreatitis in the majority of patients. Acute recurrent pancreatitis may therefore be regarded as a transition state until definite signs of chronic pancreatitis are detectable.

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Pancreatitis, Chronic; Recurrence; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Pancreatitis is usually inflammation of the pancreas without infection. Our understanding of pancreatitis has been built on autopsy studies, surgical biopsies and surrogate markers of inflammation and fibroses, including abdominal imaging techniques and pancreatic functional studies. However, the discovery that a number of different environmental factors and various genetic abnormalities are seen in patients with similar appearing pancreatitis phenotypes teaches us that end-stage pathology is not the disorder. Understanding complex associations and interactions requires that the components and their interactions be organized, stratified and functionally defined. Systems biology, in the broad sense, provides the approach and tools to define the complex mechanisms driving pathology. As the mathematics behind these pathways and mechanisms are defined and calibrated, the potential pathology of patients with early signs of disease can be predicted, and a number of patient-specific targets for intervention can be defined.

Topics: Alcoholism; Biomarkers; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Testing; Humans; Meta-Analysis as Topic; Models, Biological; Pancreas; Pancreatitis; Systems Biology; Trypsinogen

Trypsinogens and PSTI/TATI/SPINK1 are expressed, usually together, at high levels by the pancreas but also by many other normal and malignant tissues. The present review describes studies on the expression and putative functions of trypsinogens and PSTI/TATI/SPINK1 in the human body. The clinical aspects are discussed, including the correlations between expression of trypsinogens and PSTI/TATI/SPINK1 in tissues, serum, and urine of patients with pancreatitis or cancer and clinicopathological characteristics, i.e., the roles of trypsinogens and PSTI/TATI/SPINK1 in spontaneous and hereditary pancreatitis, tumor progression, and prognosis.

Topics: Biomarkers, Tumor; Humans; Neoplasms; Pancreas; Pancreatic Neoplasms; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Serum amylase remains the most commonly used biochemical marker for the diagnosis of acute pancreatitis, but its sensitivity can be reduced by late presentation, hypertriglyceridaemia, and chronic alcoholism. Urinary trypsinogen-2 is convenient, of comparable diagnostic accuracy, and provides greater (99%) negative predictive value. Early prediction of the severity of acute pancreatitis can be made by well validated scoring systems at 48 hours, but the novel serum markers procalcitonin and interleukin 6 allow earlier prediction (12 to 24 hours after admission). Serum alanine transaminase >150 IU/l and jaundice suggest a gallstone aetiology, requiring endoscopic retrograde cholangiopancreatography. For obscure aetiologies, serum calcium and triglycerides should be measured. Genetic polymorphisms may play an important role in "idiopathic" acute recurrent pancreatitis.

Topics: Acute Disease; Alanine Transaminase; Amylases; Biomarkers; Calcitonin; Calcitonin Gene-Related Peptide; Humans; Interleukin-6; Isoenzymes; Lipase; Pancreatitis; Protein Precursors; Sensitivity and Specificity; Time Factors; Trypsinogen

Acute pancreatitis is generally believed to be a disease in which the pancreas is injured by digestive enzymes that it normally produces. Most of the potentially harmful digestive enzymes produced by pancreatic acinar cells are synthesized and secreted as inactive zymogens which are normally activated only upon entry into the duodenum but, during the early stages of acute pancreatitis, those zymogens become prematurely activated within the pancreas and, presumably, that activation occurs within pancreatic acinar cells. The mechanisms responsible for intracellular activation of digestive enzyme zymogens have not been elucidated with certainty but, according to one widely recognized theory (the "co-localization hypothesis"), digestive enzyme zymogens are activated by lysosomal hydrolases when the two types of enzymes become co-localized within the same intracellular compartment. This review focuses on the evidence supporting the validity of the co-localization hypothesis as an explanation for digestive enzyme activation during the early stages of pancreatitis. The findings, summarized in this review, support the conclusion that co-localization of lysosomal hydrolases with digestive enzyme zymogens plays a critical role in permitting the intracellular activation of digestive enzymes that leads to acinar cell injury and pancreatitis.

Topics: Acute Disease; Amylases; Animals; Cathepsin B; Enzyme Activation; Enzyme Precursors; Humans; Hydrolases; Lysosomes; Pancreas; Pancreatitis; Protein Transport; Trypsinogen

There are multiple PRSS1 mutations described in hereditary pancreatitis but only a minority of these are clinically relevant. The two most frequent point mutations are in exon 2 (N29I) and exon3 (R122H), found in diverse racial populations. Both mutations result in early onset pancreatitis but the mechanism underlying this phenotype is unclear. The frequency of these mutations in such diverse populations suggests they have spontaneously occurred many times. The origin of the major mutations may be explained by gene conversions, accounting for multiple founders. The implications are discussed in terms of mechanism of action of the mutations and clinical presentation.

Topics: Gene Conversion; Genetic Predisposition to Disease; Genetic Variation; Humans; Pancreatitis; Point Mutation; Trypsin; Trypsinogen

A wide range of mutations and polymorphisms in genes that relate to pancreatic function seem to be involved in the development of pancreatitis. Some of these genetic alterations lead to disease phenotypes with unequivocal mendelian inheritance patterns, whereas others seem to act as modifier genes in conjunction with environ-mental or, as yet unidentified, genetic cofactors. This article reviews germline changes in the genes for trypsin, pancreatic secretory trypsin inhibitor, the cystic fibrosis conductance regulator, lipid metabolism proteins, inflammatory mediators for cytokines, and cathepsin B.

Topics: Carrier Proteins; Cathepsin B; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Predisposition to Disease; Germ-Line Mutation; Humans; Pancreatitis; Point Mutation; Polymorphism, Genetic; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The past decade has witnessed remarkable progress in the genetics of chronic pancreatitis. Despite these accomplishments, the understanding of the molecular mechanisms through which PRSS1 and SPINK1 mutations cause chronic pancreatitis has remained sketchy. Pancreatitis-associated gene mutations are believed to result in uncontrolled trypsin activity in the pancreas. Experimental identification of the disease-relevant functional alterations caused by PRSS1 or SPINK1 mutations proved to be challenging, however, because results of biochemical analyses lent themselves to different interpretations. This article focuses on PRSS1 mutations and summarizes the salient biochemical findings in the context of the mechanistic models that explain the connection between mutations and hereditary pancreatitis.

Topics: Carrier Proteins; Chronic Disease; Genetic Variation; Humans; Pancreatitis; Point Mutation; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The endogenous pancreatic trypsin inhibitor, SPINK, is believed to limit enzyme activity in the pancreas and reduce the risk of pancreatitis. Recently, mutations in the SPINK1 gene have been associated with development of both acute and chronic pancreatitis. In most patients with SPINK1 mutations, the genetic variants do not cause the disease independently, but may act in concert with other genetic or environmental factors. Recent studies, using mice in which the trypsin inhibitor gene has been deleted or overexpressed, provide novel insights into the role of SPINK in pancreatic development and pancreatitis.

Topics: Animals; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Enzyme Activation; Genetic Predisposition to Disease; Humans; Mice; Mice, Knockout; Pancreatitis; Point Mutation; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The number of hospitalizations in children with acute and chronic pancreatitis is increasing and accounts for significant morbidity. Acute pancreatitis is a reversible event involving diffuse inflammation of the pancreas with variable involvement of other regional tissues, remote organs, or both, whereas chronic pancreatitis is a process that produces irreversible changes in the pancreatic structure and function. Mutations in the gene encoding cationic trypsinogen have recently been identified to be associated with hereditary pancreatitis. Genetic mutations in the pancreatic secretory trypsin inhibitor and the cystic fibrosis transmembrane conductance regulator have been described to play a role in the development of pancreatitis as well. Mutations in the cytokine target genes relating to regulation of inflammation are likely to be important in determining the severity of pancreatitis. These findings, along with the advances in cell biology, have contributed to a better understanding of the pathophysiology of pancreatic diseases.

Topics: Carrier Proteins; Child; Cystic Fibrosis Transmembrane Conductance Regulator; DNA; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Polymorphism, Genetic; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Trypsin activity is properly suppressed in the pancreatic acinar cells under normal conditions. A small amount of trypsinogen is converted to active trypsin and inactivated by pancreatic secretory trypsin inhibitor (PSTI), thereby preventing damage to pancreatic acinar cells as a first line of defense. However, if trypsin activation (due to excessive stimulation of pancreatic acinar cells) exceeds the capacity of PSTI, a subsequent cascade of events leads to the activation of various proteases that damage cells. This can be interpreted as the main causative event of pancreatitis onset. Trypsin produced in and secreted from the pancreatic acinar cells activates protease activated receptor-2 (PAR-2), which is present at high densities on the luminal surfaces of pancreatic acinar cells and duct cells. Results of PAR-2 activation are the production of cytokines and the regulation of exocrine function via a negative feedback loop. Thus, the actions of trypsin, trypsin inhibitor (PSTI), and trypsin receptor (PAR-2) in the pancreas are strongly interconnected.

Topics: Animals; Carrier Proteins; DNA; Humans; Mutation; Pancreatitis; Receptor, PAR-2; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsin Inhibitors; Trypsinogen

Trypsin activity is properly suppressed by pancreatic secretory trypsin inhibitor (PSTI), which is also known as serine protease inhibitor Kazal type 1 (SPINK1), thereby preventing damage to pancreatic acinar cells as a first line of defence. However, if trypsin activation exceeds the capacity of PSTI/SPINK1, a subsequent cascade of events leads to the activation of various proteases that damage cells. Five mutations (R122H, N29I, A16V, D22G and K23R) in cationic trypsinogen and two mutations (N34S and M1T) in the PSTI/SPINK1 gene have been found to correlate significantly with the onset of pancreatitis. From analyses of hereditary pancreatitis and the phenotype of PSTI/SPINK1 (Spink3) knockout mice, we showed that the imbalance of trypsin activation and its inhibition by PSTI/SPINK1 would lead to the development of pancreatitis.

Topics: Animals; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mice; Mutation; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The discovery of PRSS 1 mutations in hereditary pancreatitis and analysis of how the genotype affects the presentation and progression of hereditary pancreatitis has led to a better understanding of the pathophysiology of the disease. Patients with hereditary pancreatitis present with symptoms at an early age and have a significant lifetime risk for the development of endocrine and exocrine insufficiency, albeit at a later stage than patients with either idiopathic or alcoholic chronic pancreatitis. There are distinct phenotypic differences between hereditary pancreatitis and with other types of pancreatitis. As many as 80% of patients with symptomatic hereditary pancreatitis have an underlying causative PRSS1 mutation; there are, however, few significant phenotypic differences between these PRSS1 mutations. TheR122H mutation is the most common PRSS1 mutation observed, and patients with the R122H mutation present earlier. This, however, does not necessarily translate into a more aggressive disease with respect to complications of chronic pancreatitis. Indeed, the age of presentation of symptoms may be a poor surrogate for predicting outcome, as inherited disorders of trypsinogen may cause subclinical attacks of pancreatitis, which ultimately lead to pancreatic destruction and dysfunction. All patients, irrespective of whether they carry a PRSS1 mutation, are at significant risk of developing pancreatic ductal adenocarcinoma. The risk appears to be insignificant below the age of 40 years, but it increases incrementally thereafter. Significantly, the risk of pancreatic cancer is not related to PRSS1 mutation type and does not appear to be related to the mode of inheritance. The role of SPINK1 mutations in modifying the expression of PRSS1mutations is unclear but appears to be of clinical importance. It is unlikely that they act as causative mutations per se, at least in the Western form of the disease. Additionally, they do not appear to have an impact on the penetrance of PRSS1 gene mutations in hereditary pancreatitis.

Topics: Chronic Disease; Humans; Mutation; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

The understanding of pathogenesis of acute and chronic pancreatitis has benefited from the progress made in genetic investigations. The discoveries of the gain of function mutations of cationic trypsinogen gene (PRSS1) and the loss of function mutations of pancreatic secretory trypsin inhibitor (SPINK 1) or other potential defects in genes that regulate pancreatic secretory function or modulate inflammatory response to pancreatic injury has changed our current concepts on the pathogenesis of pancreatitis. Genetic factors play an important role in the susceptibility to pancreatic injury, severity and evolution of inflammatory process, leading in some cases to chronic inflammation and/or fibrosis. Acute pancreatitis is viewed as an event and chronic pancreatitis as a process, sequentially linked, reflecting a complex interaction between genetic and environmental factors.

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Fibrosis; Genetic Predisposition to Disease; Genetic Testing; Humans; Inflammation; Pancreas; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

There was some recent progress in the understanding of genetic risk factors in chronic pancreatitis. Due to this progress some of the traditional views of the subject will change. Today, genetic risk factors are attributed a much more important role that in the past. The frequency and strength of mutations were higher than expected. Strong variants were the rare autosomal-dominant mutations N29I and R122H of PRSS1 (cationic trypsinogen) and homozygous N34S of SPINK1 (pancreatic secretory trypsin inhibitor). Other mutations (heterozygous N34S, CFTR) were of lower relevance but still mediate a higher risk than alcohol consumption. The course of genetically determined pancreatitis is rather mild. In the long term pancreas cancer was found in some patients but apart from non-smoking no adequate prophylactic strategy is available up to now.

Topics: Carrier Proteins; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Diabetes Mellitus; Disease Progression; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Acute pancreatitis is a common digestive disease of which the severity may vary from mild, edematous to severe, necrotizing disease. An improved outcome in the severe form of the disease is based on early identification of disease severity and subsequent focused management of these high-risk patients. However, the ability of clinicians to predict, upon presentation, which patient will have mild or severe acute pancreatitis is not accurate. Prospective systems using clinical criteria have been used to determine severity in patients with acute pancreatitis, such as the Ranson's prognostic signs, Glasgow score, and the acute physiology and chronic health evaluation II score (APACHE II). Their application in clinical practise has been limited by the time delay of at least 48 h to judge all parameters in the former two and by being cumbersome and time-consuming in the latter. Contrast-enhanced computed tomography is presently the most accurate non-invasive single method to evaluate the severity of acute pancreatitis. It cannot, however, be performed to all patients with acute pancreatitis. Therefore, considerable interest has grown in the development of reliable biochemical markers that reflect the severity of acute pancreatitis. In this article we critically appraise current and new severity markers of acute pancreatitis in their ability to distinguish between mild and severe disease and their clinical utility.

Topics: Acute Disease; C-Reactive Protein; Calcitonin; Cytokines; Health Status Indicators; Humans; Oligopeptides; Pancreatitis; Peptides; Protein Precursors; Proteins; Trypsin; Trypsinogen

The first family of hereditary pancreatitis was described in 1952. The mode of inheritance is autosomal dominant trait with an 80% of penetrance rate. Although hereditary pancreatitis is rare, this disorder has provided valuable insights in understanding the pathophysiology of pancreatitis and pancreatic cancer. The causative gene of hereditary pancreatitis was identified in 1996 through mutational analysis of genes within chromosome 7q35. Most forms of hereditary pancreatitis are caused by one of two common mutations, R122H in the third exon or N29I in the second exon of the cationic trypsinogen gene (protease serine 1, PRSS1). R122H mutation is the most common PRSS1 mutation. Additional mutations of the cationic trypsinogen gene have been described. In Korea, first family of hereditary pancreatitis with cationic trypsinogen gene mutation revealed an arginine to histidine amino acid substitution at the residue 122. Patients with hereditary pancreatitis present with symptoms at an early age and have significant risk for the development of chronic pancreatitis and pancreatic cancer. The risk of pancreatic cancer is estimated to be 53-fold higher after the age of 50 years than the general population. The risk of pancreatic cancer is not related to the type of mutation. Since hereditary pancreatitis is a strong risk factor for pancreatic cancer, it is important to establish a diagnostic criteria for diagnosis and surveillance. However, there are potential benefits, risks and limitations in genetic testing for hereditary pancreatitis. It is difficult to provide the proper treatment, but recent developments in therapeutic approaches may be helpful in caring hereditary pancreatitis. This article includes the current status, pathogenesis, clinical features, and management of hereditary pancreatitis including the aspects of pancreatic cancer.

Topics: Amino Acid Substitution; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Cytogenetic Analysis; Humans; Intercellular Signaling Peptides and Proteins; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Abdominal Pain; Alcohol Drinking; Chronic Disease; Diabetes Mellitus; Fibrosis; Gallstones; Humans; Hypercalcemia; Hyperlipidemias; Malabsorption Syndromes; Mutation; Pancreas; Pancreatitis; Risk Factors; Smoking; Trypsin; Trypsinogen

Topics: Alcohol Drinking; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Intercellular Signaling Peptides and Proteins; Mutation; Pancreatitis; Risk Factors; Smoking; Smoking Cessation; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Inflammatory disease of the pancreas falls into two major classifications: acute and chronic. Acute pancreatitis is a reversible process, whereas chronic pancreatitis produces irreversible changes in the architecture and function of the pancreas. The recent finding that mutations in the gene encoding cationic trypsinogen are associated with hereditary pancreatitis, the identification of genes that increase the risk for developing chronic pancreatitis, and advances in cell biology have contributed greatly to our understanding of the molecular mechanisms leading to pancreatitis. Although pancreatitis is less common in children than in adults, it still occurs with regularity and should be considered in any child with acute or chronic abdominal pain. The major difference between pancreatitis in children and adults lies in the etiologies and outcome of acute pancreatitis and in the etiology of chronic pancreatitis. The treatment of acute and chronic pancreatitis is similar at all ages.

Topics: Acute Disease; Animals; Carrier Proteins; Child; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Carrier Proteins; Databases, Genetic; Genetic Predisposition to Disease; Humans; Internet; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Acute Disease; Carrier Proteins; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Predisposition to Disease; Genetic Testing; Humans; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The discovery of PRSS1 mutations in hereditary pancreatitis and analysis of how the genotype affects the presentation and progression of hereditary pancreatitis has led to a better understanding of the pathophysiology of the disease. Patients with hereditary pancreatitis present with symptoms at an early age and have a significant lifetime risk for the development of endocrine and exocrine insufficiency, albeit at a later stage than patients with either idiopathic or alcoholic chronic pancreatitis. There are distinct phenotypic differences between hereditary pancreatitis and with other types of pancreatitis. As many as 80% of patients with symptomatic hereditary pancreatitis have an underlying causative PRSS1 mutation; there are, however, few significant phenotypic differences between these PRSS1 mutations. The R122H mutation is the most common PRSS1 mutation observed, and patients with the R122H mutation present earlier. This, however, does not necessarily translate into a more aggressive disease with respect to complications of chronic pancreatitis. Indeed, the age of presentation of symptoms may be a poor surrogate for predicting outcome, as inherited disorders of trypsinogen may cause subclinical attacks of pancreatitis, which ultimately lead to pancreatic destruction and dysfunction. All patients, irrespective of whether they carry a PRSS1 mutation, are at significant risk of developing pancreatic ductal adenocarcinoma. The risk appears to be insignificant below the age of 40 years, but it increases incrementally thereafter. Significantly, the risk of pancreatic cancer is not related to PRSS1 mutation type and does not appear to be related to the mode of inheritance. The role of SPINK1 mutations in modifying the expression of PRSS1 mutations is unclear but appears to be of clinical importance. It is unlikely that they act as causative mutations per se, at least in the Western form of the disease. Additionally, they do not appear to have an impact on the penetrance of PRSS1 gene mutations in hereditary pancreatitis.

Topics: Humans; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The importance of pretest information, using an accredited DNA laboratory and interpreting the genotype on behalf of the patient and their physicians is emphasized. Care with predictive testing and the strong encouragement to involve a specialist genetic counseling service is made. A similar approach to genetic testing should be used when children are involved. Because of the incomplete pickup of PRSS1 mutations, particularly of a limited mutation panel of R122H and N291 (perhaps with A16V), a diagnosis of HP cannot be ruled out by molecular genetic testing alone. The A16V mutation has a reduced penetrance, and its contribution to pancreatitis remains unclear. The advice to patients with genetic forms of pancreatitis is a strong encouragement to avoid smoking, to avoid alcohol, and to remain in contact with clinical and research groups for their follow-up and screening trials for early pancreatic cancer. The remaining issues are of how wide to cast the net of investigation in patients with unexplained pancreatitis, particularly looking for mutations in the CFTR and lower penetrance genes such as PSTI/SPINK1.

Topics: Algorithms; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Genetic Counseling; Genetic Testing; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Pregnancy; Prenatal Diagnosis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Acute Disease; Animals; Cytokines; Disease Progression; Endotoxemia; Humans; Inflammation Mediators; Leukocyte Elastase; Multiple Organ Failure; Pancreatitis; Receptor, PAR-2; Severity of Illness Index; Trypsin; Trypsinogen

Independent of the etiology, acute pancreatitis is associated with significant morbidity and the potential for mortality. In most patients, acute pancreatitis follows an uncomplicated or mild course. Recent studies in hereditary pancreatitis have clearly revealed that trypsin is the key enzyme at the onset of pancreatitis. However, there are several defense mechanisms to prevent ectopic activation of trypsin under physiological conditions. If the defense mechanisms failed or activation of trypsin occurred over defense ability, trypsin would activate other digestive enzymes and self-digestion of the pancreas would occur.

Topics: Acute Disease; Animals; Carrier Proteins; Cathepsin B; Humans; Intercellular Signaling Peptides and Proteins; Pancreas; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Serum amylase is most commonly used as a biochemical marker of acute pancreatitis (AP). But it lacks specificity. The serum lipase level is more accurate and a better marker. Serum elastase -1 level is specific and remains elevated longer, but its radioimmunoassay is not routinely used. Recently, it can be rapidly measured by latex turbidometric immunoassay with automatic analyzer. Biochemically, only CRP test is available and useful to assess severity, but its sensitivity is unacceptably low in the early course of the disease. Urinary trypsinogen activation peptide (TAP) or trypsinogen-2 is an earlier marker. Increasing knowledge of the inflammatory process in AP has led to possibly useful biochemical indicators of severity, such as cytokines, nonpancreatic synovial type group II PLA2 or granulocyte elastase.

Topics: Acute Disease; Amylases; Antigens, Neoplasm; Biomarkers; Biomarkers, Tumor; C-Reactive Protein; Cytokines; Humans; Lectins, C-Type; Leukocyte Elastase; Lipase; Nephelometry and Turbidimetry; Oligopeptides; Pancreatic Elastase; Pancreatitis; Pancreatitis-Associated Proteins; Phospholipases A; Phospholipases A2; Severity of Illness Index; Trypsin; Trypsinogen

Since the discovery of the cationic trypsinogen gene mutations in patients with hereditary pancreatitis, a variety of pancreatitis-associated gene mutations have been reported, including pancreatic secretory trypsin inhibitor and cystic fibrosis transmembrane conductance regulator. Although the patients with these mutations are rarely seen, genetic disorders inducing pancreatitis have provided us major breakthroughs to understand the molecular basis of the disease. Furthermore, the major stream in pancreatology has been evidenced in patients with hereditary pancreatitis: acute pancreatitis --> chronic pancreatitis --> pancreatic cancer. This report will focus on the pancreatitis-associated genes and the molecular mechanism of pancreatitis associating with these gene mutations.

Topics: alpha 1-Antitrypsin; Animals; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Intercellular Signaling Peptides and Proteins; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The classical feature of hereditary pancreatitis (HP) is characterized by recurrent episodes of acute pancreatitis or a priori chronic pancreatitis in several members of one family. In 1996, the identification of the first HP-associated mutation in the cationic trypsinogen gene provided a breakthrough in our understanding of the pathogenesis of chronic pancreatitis. In the following years, several different mutations in the same gene have been found in a large number of investigated families. Most intriguing, HP patients have a more than 50-fold increased risk of pancreatic ductal cancer in comparison with expected pancreatic cancers in the general population. Variants of the major intrapancreatic trypsin antagonist SPINK1 have implications for more common forms of chronic pancreatitis. Research has focussed on the SPINK1-N34S-mutation, which is closely associated with tropical, alcoholic, or "idiopathic" chronic pancreatitis. Chronic pancreatitis represents a variable part of the cystic fibrosis syndrome, which is caused by mutations in the gene coding for the cystic fibrosis transmembrane conductance regulator (CFTR). Several groups have reported an increased prevalence of CFTR mutations in patients with chronic pancreatitis of different etiology. In this review, we summarize interesting clinical and biochemical features of genetic variants in these genes which are associated with chronic pancreatitis.

Topics: Carrier Proteins; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Poland; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Adult; Chronic Disease; Disease Progression; Female; Humans; Male; Mutation; Pancreatitis; Pancreatitis, Alcoholic; Registries; Trypsin; Trypsinogen; Twins, Monozygotic

Chronic pancreatitis remains a challenging and frustrating clinical problem. In the past few years, however, advances in genetic and immunologic research have spawned new insights and approaches to chronic pancreatitis. Genetic and environmental risk assessment may help identify individuals who are likely to develop severe chronic pancreatitis early in the disease course, and allow targeted attention to reduce confounding risks and slow or prevent this problem in the future.

Topics: Chronic Disease; Humans; Models, Biological; Pancreatitis; Risk Factors; Trypsin Inhibitors; Trypsinogen

Hereditary pancreatitis is an autosomal dominant condition, which results in recurrent attacks of acute pancreatitis, progressing to chronic pancreatitis often at a young age. The majority of patients with hereditary pancreatitis express one of two mutations (R122H or N29I) in the cationic trypsinogen gene (PRSS1 gene). It has been hypothesised that one of these mutations, the R122H mutation causes pancreatitis by altering a trypsin recognition site so preventing deactivation of trypsin within the pancreas and prolonging its action, resulting in autodigestion. Families with these two mutations have been identified in many countries and there are also other rarer mutations, which have also been linked to hereditary pancreatitis. Patients with hereditary pancreatitis present in the same way as those with sporadic pancreatitis but at an earlier age. It is common for patients to remain undiagnosed for many years, particularly if they present with non-specific symptoms. Hereditary pancreatitis should always be considered in patients who present with recurrent pancreatitis with a family history of pancreatic disease. If patients with the 2 common mutations are compared, those with the R122H mutation are more likely to present at a younger age and are more likely to require surgical intervention than those with N29I. Hereditary pancreatitis carries a 40 % lifetime risk of pancreatic cancer with those patients aged between 50 to 70 being most at risk in whom screening tests may become important.

Topics: Genetic Counseling; Genetic Testing; Humans; Pancreatitis; Point Mutation; Risk Factors; Trypsin; Trypsinogen

Hereditary pancreatitis (HP) is a disease which has been discovered quite recently. The inheritance is autosomally dominant with 80% penetrance. It gives the same symptoms as acute pancreatitis in early childhood and ends up with chronic pancreatitis. In 60% of the patients, a mutation in the trypsinogen gene can be demonstrated. The remaining 40% of the HP patients are diagnosed on the basis of clinical criteria. The acute and the chronic pancreatitis are treated as usual. It is important to recognize the disease because patients with HP have a 50 times increased risk of developing pancreatic cancer. At the age of 70, 40% have developed pancreatic cancer. This risk doubles for cigarette smokers. Screening programmes for HP in order to prevent pancreatic cancer are, however, expensive and troublesome.

Topics: Acute Disease; Adult; Aged; Child; Chronic Disease; Genetic Predisposition to Disease; Genetic Testing; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

A number of genetic mutations have recently been identified that appear to be important in the development of pancreatitis. Point mutations in the cationic trypsinogen gene are capable of initiating pancreatitis. These mutations also provide important insights into the pathophysiology of acute pancreatitis and into potential connections between acute and chronic pancreatitis. Mutations in the genes encoding for the pancreatic secretory trypsin inhibitor and the cystic fibrosis transmembrane conductance regulator more likely work in concert with other genes and environmental factors in affecting disease susceptibility. Although the subject so far has received only a limited amount of study, genetic polymorphisms in a wide range of genes relating to pancreatic function and to regulation of inflammation are likely to play major roles in determining each individual's susceptibility to developing pancreatitis, and its severity if it does develop.

Topics: Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Penetrance; Peptide Fragments; Point Mutation; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Genetic changes associated with some forms of chronic pancreatitis have been recently defined. There are three genes that play a role, each with a variety of genotypes and different pathologic mechanisms and clinical correlations. Selection of the appropriate diagnostic tests requires integration of the clinical and family history and the interpretation of results has a significant impact on genetic counseling for the patient and family. The relative significance of some variant alleles is still under investigation as they are common in the population and show low penetrance. Knowledge of the pathophysiology of each abnormal allele could lead the way towards more specific therapeutic options in the future.

Topics: Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Counseling; Genetic Testing; Humans; Molecular Diagnostic Techniques; Pancreas; Pancreatitis; Serine Proteinase Inhibitors; Trypsin; Trypsinogen

Topics: Acute Disease; Humans; Pancreatitis; Serine Proteinase Inhibitors; Trypsin; Trypsinogen

Topics: DNA, Mitochondrial; Enzymes; Humans; Infant; Infant, Newborn; Mutation; Pancreas; Pancreatic Diseases; Pancreatitis; Trypsinogen

Pancreatitis has a proven genetic basis in a minority of patients.. Review of the literature on genetics of pancreatitis.. Ever since the discovery that in most patients with hereditary pancreatitis a mutation in the gene encoding for cationic trypsinogen (R122H) was found that results in a gain of trypsin function', many other mutations in the cationic trypsinogen gene, as well as in the gene encoding for pancreatic secretory trypsin inhibitor, have been found in patients with chronic pancreatitis. Furthermore, mutations in other genes, like the mucoviscoidosis-gene encoding for a chloride channel, and in genes encoding for enzymes involved in the metabolism of ethanol, have been linked to chronic pancreatitis. This article reviews the highlights that have been achieved in this field of pancreatic research.. Recent data suggest that genetics may play a role in the pathogenesis of pancreatitis.

Topics: Chronic Disease; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

Topics: Cystic Fibrosis; Humans; Pancreatitis; Point Mutation; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Acute pancreatitis is one of the major complications of ERCP. It is of paramount importance that we accurately identify which patients will go on to develop post-ERCP pancreatitis. As most ERCPs are performed on an outpatient basis, early evaluation can allow safe discharge of the majority of patients who will not develop post-ERCP pancreatitis or develop only mild symptoms that will be self-limited. Alternatively, early detection of those patients who will go on to develop moderate or severe post-ERCP pancreatitis can guide decisions regarding hospital admission and aggressive management and can help direct the use of targeted therapies that have the potential to prevent or mitigate pancreatic inflammation. Thus, significant efforts have focused on trying to identify predictors of post-ERCP pancreatitis. These parameters can be organized into three categories of tests: 1) pancreatic enzymes as markers of pancreatic injury: serum amylase/urine amylase; 2) markers of proteolytic activation: trypsinogen, trypsinogen activation peptide; 3) markers of systemic inflammation: C-reactive protein, various interleukins such as IL-6 and IL-10. A serum amylase level greater than 4-5 times the upper reference limit in conjunction with clinical symptoms has been shown to be an accurate and reliable predictor of post-ERCP pancreatitis. However, the exact timing and level of amylase elevation remains debatable. Urine testing of amylase and trypsinogen-2 in post-ERCP patients has also been shown to be highly sensitive and specific for detecting pancreatitis. The main advantage of these urinary markers is that they are available as rapid dipstick tests. Serum trypsinogen-2 levels have also been studied in post-ERCP pancreatitis patients; high levels seem to correlate with severity of disease. Among the markers of systemic inflammation, serum CRP is an accurate and readily available laboratory test for predicting severity of post-ERCP pancreatitis, but it appears to be helpful at 24-48 hours and, therefore, is not an early marker. Several other markers remain investigational and have not yet found wide clinical applicability.

Topics: Acute Disease; Amylases; Biomarkers; C-Reactive Protein; Cholangiopancreatography, Endoscopic Retrograde; Enzyme Activation; Humans; Inflammation; Interleukins; Oligopeptides; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

Topics: alpha 1-Antitrypsin; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Risk Factors; Serine Proteinase Inhibitors; Trypsin; Trypsinogen

Topics: alpha 1-Antitrypsin; Child; Chromosomes, Human, Pair 12; Chromosomes, Human, Pair 14; Chromosomes, Human, Pair 5; Chromosomes, Human, Pair 7; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatitis; Prevalence; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Pancreatitis is rightly the most feared complication of endoscopic retrograde cholangiopancreatography (ERCP). Ten percent to 15% of cases of post-ERCP pancreatitis (PEP) are severe by clinical and radiologic criteria. Such cases carry significant morbidity and mortality and are responsible for the vast majority of ERCP-related deaths. The prediction and prevention of PEP have been of great interest to endoscopists since the introduction of ERCP 30 years ago. Prediction and diagnosis of PEP have become more accurate with the widespread availability of serum amylase estimation. A variety of cytokines (eg, interleukin -1, IL-6, and IL-8) and acute phase reactants (eg, C-reactive protein) are also elevated in the serum in acute pancreatitis, and these form the basis of evolving tests for PEP. Urine testing (for amylase) in acute pancreatitis is obsolete, but it may soon undergo a revival in the form of a rapid (3-minute) dipstick test for trypsinogen-2, a sensitive and specific test for this disease. The prevention of PEP takes multiple forms. The following steps are recommended for clinicians: 1) avoid ERCP when other, less invasive or noninvasive imaging tests can do the job (eg, CT or magnetic resonance imaging); 2) avoid high-risk (of PEP) procedures, such as needle-knife papillotomy, balloon dilation of the biliary sphincter, and pancreatic sphincterotomy, and take steps to reduce risk when these procedures are unavoidable; 3) ensure that those who perform ERCP have adequate training and experience; and 4) consider pharmacologic intervention. Despite a depressing catalog of drug interventions that have failed over the years (eg, antihistamines, anticholinergics, and corticosteroids), three agents have recently shown promise: somatostatin; its octapeptide analogue, octreotide; and gabexate mesylate, a protease inhibitor.

Topics: Acute Disease; Acute-Phase Proteins; Amylases; Biomarkers; Calcitonin; Cholangiopancreatography, Endoscopic Retrograde; Contrast Media; Gabexate; Hormones; Humans; Interleukin-10; Interleukins; Octreotide; Pancreatitis; Protein Precursors; Risk Assessment; Serine Proteinase Inhibitors; Somatostatin; Trypsinogen

The diagnosis of acute pancreatitis depends on a combination of clinical assessment and laboratory testing. Although the serum amylase is the cornerstone laboratory test used in establishing the diagnosis of acute pancreatitis, there are limitations in the sensitivity and specificity that may be important for the clinician to recognize. The serum lipase level may be especially useful in patients with alcohol-induced acute pancreatitis. A new urinary test strip that uses trypsinogen-2 may have a role in establishing the diagnosis of acute pancreatitis. In addition, several new laboratory tests and new interpretations of old laboratory tests may assist in establishing the etiology and severity of acute pancreatitis. This review summarizes important aspects of standard laboratory tests and novel laboratory approaches in establishing the diagnosis, etiology, and severity of acute pancreatitis.

Topics: Acute Disease; Alanine Transaminase; Amylases; Cholelithiasis; Humans; Lipase; Pancreatitis; Pancreatitis, Alcoholic; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen

Topics: Acute Disease; Animals; Antigens, Neoplasm; Biomarkers, Tumor; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Disease Models, Animal; Genetic Diseases, Inborn; Genetic Heterogeneity; Humans; Lectins, C-Type; Mice; Molecular Biology; Mutation; Pancreatitis; Pancreatitis-Associated Proteins; Proteins; Rats; Risk Factors; Trypsinogen

GENETIC RISK FACTORS: Recently, several genetic risk factors for chronic pancreatic diseases were found. In patients with chronic pancreatitis several mutations of the cationic trypsinogen were identified. In the majority of these subjects an autosomal dominant disease was observed. Mutations in the pancreatic secretory trypsin inhibitor (SPINK 1) were found in 20% of subjects with idiopathic, in 5% of those with alcoholic chronic and in 50% of those with tropical pancreatitis. Further variants were identified in CFTR (Cystic Fibrosis Transmembrane Conductance Regulator), a chlorid transporter initially identified a disease-causing molecule in cystic fibrosis. In approximately 20-25% of the patients with chronic pancreatitis a mutation of one of these genes can be found.. A genetic investigation is useful when there is evidence for a family history of chronic pancreatitis and, at absence of typical risk factors, in patients with onset of disease earlier than 35-40 years of age.. In addition to alcohol abstinence the patients should be advised not to smoke. National and international registers as well as self-aid groups exist.

Topics: Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Genetic Predisposition to Disease; Humans; Pancreatitis; Pancreatitis, Alcoholic; Risk; Trypsin; Trypsinogen

Topics: alpha-Amylases; Chronic Disease; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Plant Proteins; Trypsin Inhibitors; Trypsinogen

In the last few years, several genes have been identified as being associated with hereditary and idiopathic chronic pancreatitis (CP), i.e. PRSS1, CFTR and SPINK1. In this study, we investigated 164 unrelated children and adolescents with CP for mutations in disease-associated genes by direct DNA sequencing, SSCP, RFLP and melting curve analysis. In 15 patients, we detected a PRSS1 mutation (8 with A16V, 5 with R122H, 2 with N29I), and in 34 patients, a SPINK1 mutation (30 with N34S, 4 with others). SPINK1 mutations were predominantly found in patients without a family history (29/121). Ten patients were homozygous for N34S, SPINK1 mutations were most common in 'idiopathic' CP, whereas patients with 'hereditary' CP predominantly showed a PRSS1 mutation (R122H, N29I). In patients without a family history, the most common PRSS1 mutation was A16V (7/121). In conclusion, our data suggest that CP may be inherited in a dominant, recessive or multigenetic manner as a result of mutations in the above-mentioned or as yet unidentified genes. This challenges the concept of idiopathic CP as a nongenetic disorder and the differentiation between hereditary and idiopathic CP. Therefore, we propose to classify CP as either 'primary CP' (with or without a family history) or 'secondary CP' caused by toxic, metabolic or other factors.

Topics: alpha 1-Antitrypsin; Child; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatitis; Serine Proteinase Inhibitors; Trypsin; Trypsinogen

Pancreatitis-associated gene mutations have been reported in patients with hereditary pancreatitis and idiopathic pancreatitis in the Caucasian population and involve the cationic trypsinogen gene, the pancreatic secretory trypsin inhibitor gene and the cystic fibrosis transmembrane conductance regulator gene. In the Japanese population, mutational screening analyses of these genes have shown several mutations. The present study reviews previous reports from Japan in order to evaluate the racial specificity of pancreatitis-associated gene mutations.

Topics: Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Japan; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

This study attempts to identify the biochemical alterations in human cationic trypsinogen and trypsin caused by the hereditary pancreatitis-associated mutations Arg117-->His and Asn21-->Ile.. Recombinant wild-type and mutant human cationic trypsinogens were expressed in Escherichia coli and purified to homogeneity, and trypsin autolysis and trypsinogen autoactivation were characterized.. Both mutations significantly enhanced the autoactivation of human cationic trypsinogen. In addition, the Arg117-->His mutation inhibited autocatalytic inactivation of trypsin, while the Asn21-->Ile mutation had no such effect.. The findings support the notion that enhanced trypsinogen activation in the pancreas is the common initiating step in hereditary pancreatitis, whereas trypsin stabilization plays a role in cases associated with the Arg117-->His mutation.

Topics: Autolysis; Catalysis; Enzyme Activation; Gene Expression Regulation; Genetic Vectors; Humans; Kinetics; Pancreatitis; Recombinant Proteins; Trypsin; Trypsinogen

Topics: Abdominal Pain; Acute Disease; Amylases; Diagnostic Imaging; Humans; Lipase; Pancreatitis; Trypsinogen

Hereditary pancreatitis is an uncommon cause of chronic pancreatitis in Western society. It should be suspected when chronic pancreatitis presents in young adults. The diagnosis is made when chronic pancreatitis is present in several members of the same family who are determined not to have other risk factors for chronic pancreatitis. Molecular research focusing on mutations in the trypsinogen gene has uncovered the genetic defects associated with hereditary pancreatitis, and this knowledge has suggested the possible pathophysiologic mechanism of this disease. Because patients with hereditary pancreatitis develop their disease early in life they are very likely to require treatment for complications. As in patients with chronic pancreatitis of other etiologies those with hereditary pancreatitis should be treated medically for acute exacerbations. When complications occur or when the disease causes intractable pain surgery is recommended. Surgical therapy is tailored to the patient's pancreatic anatomy based on endoscopic retrograde cholangiopancreatography or CT scan. The two patients described in this report underwent successful longitudinal pancreaticojejunostomy (Puestow procedure) with good results. Finally it has been shown that patients with hereditary pancreatitis are at increased risk for developing pancreatic adenocarcinoma. Although not widely used pancreatic cancer screening programs have been suggested for surveillance of these patients.

Topics: Abdominal Pain; Adult; Chronic Disease; Female; Humans; Male; Mutation; Pancreaticojejunostomy; Pancreatitis; Risk Factors; Trypsinogen

Topics: Acute Disease; Alcoholism; Apoptosis; Cathepsin B; Chemokines; Cholelithiasis; Cytokines; Humans; Necrosis; Oxidative Stress; Pancreatitis; Regeneration; Risk Factors; Trypsin; Trypsinogen

Topics: Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Since the identification in 1996 of a "gain of function" missense mutation, R122H, in the cationic trypsinogen gene (PRSS1) as a cause of hereditary pancreatitis, continued screening of this gene in both hereditary and sporadic pancreatitis has found more disease-associated missense mutations than expected. In addition, functional analysis has yielded interesting findings regarding their underlying mechanisms resulting in a gain of trypsin. A critical review of these data, in the context of the complicated biogenesis and complex autoactivation and autolysis of trypsin(ogen), highlights that PRSS1 mutations cause the disease by various mechanisms depending on which biochemical process they affect. The discovery of these mutations also modifies the classical perception of the disease and, more importantly, reveals fascinating new aspects of the molecular evolution and normal physiology of trypsinogen. First, activation peptide of trypsinogen is under strong selection pressure to minimize autoactivation in higher vertebrates. Second, the R122 primary autolysis site has further evolved in mammalian trypsinogens. Third, evolutionary divergence from threonine to asparagine at residue 29 in human cationic trypsinogen provides additional advantage. Accordingly, we tentatively assign, in human cationic trypsinogen, the strongly selected activation peptide as the first-line and the R122 autolysis site as the second-line of the built-in defensive mechanisms against premature trypsin activation within the pancreas, respectively, and the positively selected asparagine at residue 29 as an "amplifier" to the R122 "fail-safe" mechanism.

Topics: Amino Acid Sequence; Animals; Autolysis; Binding Sites; Enzyme Activation; Enzyme Stability; Evolution, Molecular; Humans; Molecular Sequence Data; Mutation, Missense; Pancreatitis; Selection, Genetic; Trypsin; Trypsinogen

For a long time the pathogenesis of pancreatitis has remained enigmatic. Recent developments in cellular and molecular biology, however, have provided a tremendous research impetus and some of its mysteries are finally being disclosed. This review discusses the implications of the discovery of the disease gene in hereditary pancreatitis and outlines recent advances in our understanding of the mechanism and site of trypsinogen activation and the role of immunocytes and cytokines in acute pancreatitis. With respect to chronic pancreatitis, this review focuses on its association with mutations in the cystic fibrosis conductance regulator gene and the mechanisms of pancreatic fibrosis. These advances in our knowledge of the pathogenesis of the disease, together with emerging biotechnological techniques, will boost the development of future therapies aimed at strategically targeting key pathophysiological processes involved in acute and chronic pancreatitis.

Topics: Acute Disease; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Cytokines; Humans; Mutation; Pancreatitis; Trypsinogen

The recent genetic discoveries in CP support the hypothesis that inappropriate intrapancreatic activation of zymogens by trypsin results in autodigestion and pancreatitis. Two different protective mechanisms prevent activation of the pancreatic digestive enzyme cascade. First, SPINK1 inhibits up to 20% of potential trypsin activity and, second, trypsin itself activates trypsin-like enzymes readily degrading trypsinogen and other zymogens. Pancreatitis may therefore be the result of an imbalance between proteases and their inhibitors within the pancreatic parenchyma. The discovery of PRSS1 mutations in families with CP was the first breakthrough in the understanding of the underlying genetic mechanisms. Enhanced trypsinogen activation may be the common initiating step in pancreatitis caused by these mutations. The discovery of SPINK1 mutations underlines the importance of the protease inhibitor system in the pathogenesis of CP. Thus, gain-of-function in the cationic trypsinogen resulting in an enhanced autoactivation, or loss-of-function mutations in SPINK1 leading to decreased inhibitory capacity, may similarly disturb the delicate intrapancreatic balance of proteases and their inhibitors. The recent findings of SPINK1, CFTR, and PRSS1 mutations in CP patients without a family history have challenged the concept of idiopathic CP as a non-genetic disorder and the differentiation between HP and ICP. There is a clear mode of autosomal dominant inheritance for some mutations (R122H, N291, possibly MIT), whereas the inheritance pattern (autosomal recessive, complex, or modifying) of other mutations (A16V, N34S) is controverted or unknown. The lack of mutations in the above-mentioned genes in many patients suggests that CP may also be caused by genetic alterations in yet unidentified genes. Evaluation of CP patients without an obvious predisposing factor, e.g. alcohol abuse, should include genetic testing even in the absence of a family history of pancreatitis. Finally, identification of further disease-causing genes will create a better understanding of pathogenesis and may help to develop specific preventive and therapeutic strategies.

Topics: Chronic Disease; Cystic Fibrosis; Genetic Predisposition to Disease; Genetic Testing; Genotype; Humans; Mutation; Pancreatitis; Polymorphism, Genetic; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Induction of acute pancreatitis follows a uniform mechanism independent of the different etiologic factors such as gallstones, alcohol, ischemia, hyperlipidemia, hypercalcemia, hereditary and others. Each cause seems to affect primarily the acinar cell, resulting in premature intracellular activation of trypsinogen and other digestive enzymes. Activated enzymes and oxygen free radicals injure the acinar cell and cause a release of cytokines and vasoactive mediators, attract inflammatory cells and activate the vascular endothelium as well as the expression of adhesion molecules. The disturbance of the pancreatic microcirculation induces a progression from edematous to necrotizing pancreatitis independent of the early intracellular events, including protease activation. Specific therapy must be directed towards microperfusion failure as a secondary pathogenetic step, since the initial enzyme activation and cytokine release is irreversible by the time of clinical presentation. In experimental designs comparable to the clinical situation the following therapeutic principles have proven beneficial: increase of blood fluidity by dextran, inhibition of leukocyte-endothelium interaction by ICAM-1 antibodies, and blockade of local vasoconstriction by endothelin-receptor antagonists.

Topics: Acute Disease; Endopeptidases; Enzyme Activation; Humans; Pancreas; Pancreatitis; Reactive Oxygen Species; Trypsinogen

Topics: Acute Disease; Animals; Cathepsin B; Enzyme Activation; Enzyme Precursors; Pancreas; Pancreatitis; Rats; Trypsinogen

Autodigestion by proteolytic enzymes is thought to represent the critical mechanism by which acute pancreatitis is initiated. Where and why pancreatic proteases, which are physiologically stored and secreted as inactive precursor zymogens, are activated within the pancreas has remained controversial. Here we present data which indicate that: the lysosomal protease cathepsin B can activate trypsinogen in vitro in a manner that is similar to trypsinogen activation by enterokinase; that cathepsin B colocalizes with trypsinogen in the secretory compartment of the rat pancreas and of the human pancreas; that trypsinogen activation begins in a secretory compartment that is distinct from mature zymogen granules; and that the inhibition of cathepsin B can either increase or decrease premature trypsinogen activation depending on the concentration of the inhibitor, its specificity and its site of action in the pancreatic acinar cell. These observations elucidate some of the complex relations between cysteine and serine proteases in the pancreas with respect to their mechanisms of activation, their subcellular sites of action, and their possible role in the onset of pancreatitis.

Topics: Acute Disease; Animals; Cathepsin B; Ceruletide; Cytoplasmic Granules; Dogs; Enteropeptidase; Enzyme Activation; Humans; Lysosomes; Mice; Pancreas; Pancreatitis; Rats; Trypsin; Trypsinogen

Advances in molecular genetics have provided the powerful tools necessary to identify the key molecules and mechanisms that underly the disease process. Continued work in this area promises to reveal new insights as new disease genes are discovered. This article focuses on the insights into the cause of acute and chronic pancreatitis gained by investigation of the HP genes, the diagnosis of the known mutations, the fascinating observation of nonpenetrance, and a look at future directions.

Topics: Acute Disease; Amino Acid Sequence; Chronic Disease; Chymotrypsinogen; DNA Mutational Analysis; Genetic Predisposition to Disease; Genetic Testing; Humans; Molecular Sequence Data; Pancreatitis; Penetrance; Trypsinogen

Acute pancreatitis is a disorder that has numerous causes and an obscure pathogenesis. Bile duct stones and alcohol abuse together account for about 80% of acute pancreatitis. Most episodes of biliary pancreatitis are associated with transient impaction of the stone in the ampulla (that causes obstruction of the pancreatic duct, with ductal hypertension) or passage of the stone though and into the duodenum. Other causes of acute pancreatitis are various toxins, drugs, other obstructive causes (such as malignancy or fibrotic sphincter of Oddi), metabolic abnormalities, trauma, ischemia, infection, autoimmune diseases, etc. In 10% of cases of acute pancreatitis, no underlying cause can be identified; this is idiopathic pancreatitis. Occult biliary microlithiasis may be the cause of two thirds of the cases of "idiopathic" acute pancreatitis. Intra-acinar activation of trypsinogen plays a central role in the pathogenesis of acute pancreatitis, resulting in subsequent activation of other proteases causing the subsequent cell damage. Ischemia/reperfusion injury is increasingly recognized as a common and important mechanism in the pathogenesis of acute pancreatitis and especially in the progression from mild edematous to severe necrotizing form. Increased intracellular calcium concentration also mediates acinar cell damage. Oxygen-derived free radicals and many cytokines (e.g., interleukin [IL]-1, IL-6, IL-8, tumor necrosis factor-alpha, platelet activating factor) are considered to be principal mediators in the transformation of acute pancreatitis from a local inflammatory process into a multiorgan illness.

Topics: Acute Disease; Autoimmune Diseases; Child; Cholelithiasis; Female; Humans; Hypercalcemia; Male; Pancreas; Pancreatic Neoplasms; Pancreatitis; Pancreatitis, Alcoholic; Pregnancy; Reperfusion Injury; Trypsinogen

Hereditary pancreatitis (HP) is an autosomal dominant disease. Two heterozygous missense mutations, R122H (R117H) and N29I (N21I), in the cationic trypsinogen gene have been clearly associated with HP. The 'self-destruct' model proposed for the R122H mutation is discussed in connection with the existing theory of pancreatitis, and the basic biochemistry and physiology of trypsinogen, with particular reference to R122 as the primary autolysis site of the cationic trypsinogen. Two different genetic mechanisms are identified which cause the R122H mutation, and gene conversion is the likely cause of the N29I mutation. A unifying model, which highlights an indirect impairment on the R122 autolysis site is hypothesised for the N29I mutation. Possible predisposition to pancreatitis by additional DNA variants in the gene, such as the A16V signal peptide cleavage site mutation and the K23R activation peptide cleavage site mutation is suspected, but not proven. Evidence of genetic heterogeneity of HP is reviewed and cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations detected in HP families are re-evaluated. Finally, large scale association studies are expected to clarify the additional variants' role in pancreatitis and to identify new HP genes.

Topics: Amino Acid Sequence; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Diseases, Inborn; Humans; Molecular Sequence Data; Mutation, Missense; Pancreatitis; Sequence Alignment; Trypsinogen

Mutations of the cationic trypsinogen gene have been detected in hereditary pancreatitis. This article reviews current understanding of their function and clinical significance.. An unrestricted Medline search was conducted using the key words hereditary pancreatitis and 'cationic trypsinogen . Additional material was obtained from references cited in original papers and recently published abstracts of meetings.. Cationic trypsinogen mutations have been identified in most, but not all, families with hereditary pancreatitis. This confirms existing evidence that premature trypsinogen activation plays a central role in the pathogenesis of human pancreatitis. Patients currently clinically defined as having hereditary pancreatitis should be screened for the presence of cationic trypsinogen mutations. A subgroup of patients with non-hereditary pancreatitis may also benefit from being screened for these mutations. Patients with hereditary pancreatitis should be entered into prospective, multicentre trials investigating secondary screening for pancreatic cancer. Gene therapy for hereditary pancreatitis is beyond current technological capability but remains a future therapeutic prospect for this often debilitating condition.

Topics: Acute Disease; Cations; Chronic Disease; Humans; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsinogen

Hereditary pancreatitis is an autosomal dominant form of chronic pancreatitis. It presents with recurrent attacks of acute pancreatitis, usually starting in early childhood. The attacks may vary from mild abdominal pain to pancreatic necrosis, splenic vein thrombosis, pseudocysts and death. Ultimately chronic pancreatitis ensues with unrelenting pain, calcifications, endocrine and exocrine dysfunction. The penetrance is estimated at 80%. With the use of genetic linkage analysis the gene for hereditary pancreatitis was placed on the long arm of chromosome 7 (7q35). Mutational analysis identified cationic trypsinogen as the disease gene. Cationic trypsinogen mutations are thought to result in resistance of this molecule to autolysis.

Topics: Adult; Child, Preschool; Chromosomes, Human, Pair 7; Chronic Disease; Diagnosis, Differential; Genetic Linkage; Genetic Predisposition to Disease; Humans; Mutation; Netherlands; Pancreatitis; Penetrance; Trypsin; Trypsinogen

Topics: Acute Disease; Chronic Disease; Humans; Mutation; Pancreatitis; Trypsinogen

Hereditary pancreatitis is a rare condition characterized by acute and chronic pancreatitis transmitted in an autosomal dominant fashion. There also is an epidemiologic link to pancreatic cancer in some affected families. Failure of a secondary brake mechanism responsible for inactivation of prematurely activated cationic trypsin in acinar cells seems to be the fundamental defect in type I hereditary pancreatitis (R117H cationic trypsin), and also may explain the pathogenesis of type II hereditary pancreatitis (N211 cationic trypsin). The diagnosis is made based on clinical history and, in certain cases, by molecular diagnostic testing for these gene defects. Medical management of acute and chronic hereditary pancreatitis currently does not differ from that of nonhereditary AP. As in nonhereditary pancreatitis, the surgical approach must be tailored to the individual problem, with an understanding that disease restricted to the head of the gland is atypical and that residual acinar tissue continues to drive the disease state. Although diagnosis and management of pancreatic adenocarcinoma are similar in this cohort, the increased age-accumulated risk suggests that thoughtful screening protocols eventually may be clinically and cost-effective.

Topics: Adenocarcinoma; Chromosomes, Human, Pair 7; Female; Humans; Male; Pancreatic Neoplasms; Pancreatitis; Point Mutation; Trypsinogen

Hereditary pancreatitis is an unusual form of acute and chronic pancreatitis that is usually associated with two specific mutations in the cationic trypsinogen gene. The extensive information available on the biochemistry, cell biology, and molecular biology of cationic trypsinogen and its gene provides the groundwork for development of a variety of therapeutic strategies, including gene therapy. Several features of this disease, however, make gene therapy unlikely in the near future. Further research using new models, including transgenic animals, is required before breakthroughs in therapy can be expected.

Topics: Amino Acid Sequence; Animals; Chymotrypsinogen; Genetic Therapy; Humans; Molecular Sequence Data; Mutation; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Despite the recent development of medical imaging technology, chronic pancreatitis can only be diagnosed when the disease is fully established. This is due to the lack of specific and sensitive markers for this disease. The discovery of mutations in the cationic trypsinogen gene in patients with hereditary pancreatitis and a high incidence of mutations in the cystic fibrosis transmembrane conductance regulator gene in patients with chronic pancreatitis might be important clues to understanding the molecular mechanisms of this disease. The interaction between ethanol and ion channels might be the missing link between alcohol ingestion and chronic pancreatitis.

Topics: Adult; Aged; Alcoholism; Amino Acid Substitution; Animals; Biomarkers; Chloride Channels; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Dogs; Ethanol; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Models, Biological; Pancreatitis; Point Mutation; Protein Conformation; Trypsin; Trypsinogen

The recent discovery that mutations in the trypsinogen gene are responsible for acute and chronic pancreatitis, and that patients with hereditary pancreatitis are at great risk for pancreatic cancer, has opened the door to understanding many aspects of pancreatic disease. This review focuses on the clinical presentation of hereditary pancreatitis, the mechanism of disease, and implications of this disease on understanding acute and chronic pancreatitis.

Topics: Acute Disease; Amino Acid Substitution; Chronic Disease; Genetic Predisposition to Disease; Humans; Mutation, Missense; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Hereditary pancreatitis is an unusual form of acute and chronic pancreatitis with a familial predisposition. Recently, the gene mutations causing most cases of hereditary pancreatitis have been identified in the cationic trypsinogen gene. The known mutations are trypsinogen R117H and N211. These may predispose to acute pancreatitis by eliminating one of the fail-safe mechanisms used by the pancreas to eliminate prematurely activated trypsin. Accumulation of active trypsin mutants are hypothesized to initiate a digestive enzyme activation cascade in the pancreatic acinar cells leading to autodigestion, an intense inflammatory response, and acute pancreatitis. The observation that these patients also develop typical chronic pancreatitis and may later develop pancreatic cancer provides strong evidence that these conditions are linked. Knowledge of the pathophysiological conditions leading to acute and chronic pancreatitis and the development of a transgenic mouse expressing the mutant human trypsinogen genes will provide directions and tools necessary for the effective treatment or prevention of this human disease.

Topics: Animals; Chronic Disease; Disease Models, Animal; Genetic Diseases, Inborn; Humans; Mice; Pancreatic Neoplasms; Pancreatitis; Prognosis; Sensitivity and Specificity; Trypsinogen

Topics: Acute Disease; Animals; Cathepsin B; Ceruletide; Disease Models, Animal; Enzyme Activation; Humans; In Vitro Techniques; Lysosomes; Pancreas; Pancreatitis; Trypsinogen

Acute pancreatitis is a rather common abdominal disorder. In most patients the disease is mild, but about 20% of cases develop a severe necrotizing form of the disease with complications. In an emergency setting, the diagnosis of acute pancreatitis remains problematic and several patients with severe disease are diagnosed only at autopsy. Measurements of amylase or lipase are the principal laboratory methods for diagnosing acute pancreatitis. However, their sensitivity and specificity are generally considered unsatisfactory. Recent advances in the knowledge of the pathogenesis of acute pancreatitis and advances in laboratory technology have revealed new diagnostic possibilities. Especially assays based on trypsin pathophysiology have brought new alternatives for diagnostics and severity grading of the disease. Additionally, development of phospholipase A2 determinations and discovery of a new pancreatic protein, pancreatitis-associated protein, are very interesting. This article summarizes the value of new methods in the laboratory diagnostics of acute pancreatitis.

Topics: Acute Disease; Acute-Phase Proteins; Antigens, Neoplasm; Biomarkers, Tumor; Clinical Enzyme Tests; Humans; Lectins, C-Type; Pancreatitis; Pancreatitis-Associated Proteins; Phospholipases A; Phospholipases A2; Trypsinogen

There is no golden standard for the diagnosis of acute pancreatitis (AP). The diagnosis is currently based on clinical presentation, measurement of released pancreatic enzymes and imaging studies. Serum/urinary amylase, lipase and trypsinogen-2 dipstick are the most applicable methods in the clinical practice largely because of their simple, rapid, inexpensive and readily available assay methods. In addition to the clinical picture, inflammatory markers (CRP) or contrast enhanced CT can be used to assess the severity of acute pancreatitis. Multifactorial scoring systems (Ranson's prognostic signs, APACHE II, MOF-score) may be too cumbersome for clinical practice. Patient history, determination of AST, bilirubin and alkaline phosphatase levels as well as imaging studies such as ultrasonography and ERCP can be used to distinguish between biliary and non-biliary origin of the disease.

Topics: Acute Disease; Amylases; Cholangiopancreatography, Endoscopic Retrograde; Diagnosis, Differential; Humans; Lipase; Pancreatitis; Prognosis; Severity of Illness Index; Tomography, X-Ray Computed; Trypsin; Trypsinogen

Clinical assessment of acute pancreatitis by experts is as accurate as any of the individual approaches which have been recommended. What is important in a hospital setting is for one or more of these systems to be applied in individual hospitals so that forewarning is given, especially to the less experienced clinicians, of the patient who is likely to run into difficulties and requires high dependency or intensive care. One practical approach which can be personally recommended is to employ the Glasgow scoring system plus C-reactive protein levels and also to take into account body mass index. Any patient with three positive Glasgow factors, or CRP > 150 mg/l or BMI > 30 kg/m2 has severe acute pancreatitis. More refined systems may ultimately be developed but we are still some way from a single substance in blood or urine being easily and cheaply measured and representing an accurate prognostic indicator of severe acute pancreatitis. Part of the journey has been completed but there is still considerable potential to make the rest of the journey an improvement for both clinicians and patients.

Topics: Acute Disease; Adult; Aged; APACHE; C-Reactive Protein; Humans; Interleukin-6; Oligopeptides; Pancreatitis; Prognosis; Severity of Illness Index; Tomography, X-Ray Computed; Trypsinogen

Topics: Acute Disease; Chronic Disease; Computer Simulation; Humans; Models, Biological; Mutation; Pancreatitis; Trypsinogen

Important advances in the understanding of pancreatic diseases have taken place through the application of molecular methods in the study of the inherited form of pancreatitis and pancreas cancer. Mutations of the cationic trypsinogen gene have been found to be causative for hereditary pancreatitis with important implications for the molecular pathogenesis of acute and chronic pancreatitis. A variety of cancer syndromes involving the P16 and BRCA2 genes, for example, also lead to pancreatic cancer, but the gene responsible for familial pancreatic cancer has not been identified so far. The establishment of a European Registry of Hereditary Pancreatitis and Pancreatic Cancer (EUROPAC) will facilitate future developments.

Topics: Family Health; Humans; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Topics: Adult; Diabetes Mellitus, Type 1; Humans; Pancreas Transplantation; Pancreatic Diseases; Pancreatitis; Protease Inhibitors; Radioimmunoassay; Trypsin; Trypsinogen

Topics: Acute Disease; Amylases; Humans; Isoenzymes; Lipase; Pancreatitis; Trypsin; Trypsinogen

Topics: Acute Disease; Blood Coagulation; Disseminated Intravascular Coagulation; Humans; Pancreas; Pancreatitis; Pulmonary Edema; Shock; Trypsin; Trypsinogen

Oedematous pancreatitis is pancreatic acinar cell damage with leakage into the peritoneal cavity and circulation of the inactive zymogens of digestive enzymes and active amylase and lipase. Pancreatic oedema and intra-abdominal fat necrosis occur. Necrotising pancreatitis is pancreatic acinar cell damage accompanied by the specific conversion of trypsinogens to trypsins, at a rate, and on a scale, sufficient to overwhelm local defences. Rapid release of the whole spectrum of activated pancreatic enzymes leads to necrosis of parts of the pancreas and blood vessels, and the disseminated enzyme-mediated damage which characterises the molecular pathology of the established severe disease. Chronic pancreatitis, although less well understood, is also associated with trypsinogen activation within the gland. Two mechanisms have emerged as initiators of trypsinogen activation, lysosomal cathepsins and bile-borne enterokinase. Chemotherapeutic strategies against disease initiation include preparation of synthetic enterokinase and Cathepsin B inhibitors. Chemotherapeutic strategies against second-stage mediation of multi-organ damage in the disease, include oligopeptide or organic functionalities with novel catalytic site-directed moieties (such as fluoromethyl ketones) suitable for in vivo use and the specific inhibition of the relevant range of enzymes in complex with alpha 2-macroglobulin. Interference with pancreatic enzyme biosynthesis using proteolysis-resistant constructs mimicking receptor-binding domains of inhibitor peptide hormones as well as inhibitors of pancreatic signal peptidase are promising additional chemotherapeutic approaches worthy of active investigation.

Topics: Acute Disease; Cathepsin B; Cathepsins; Enteropeptidase; Enzyme Activation; Enzyme Inhibitors; Humans; Pancreas; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Acute Disease; Chronic Disease; Cytoplasmic Granules; Enzyme Activation; Enzyme Precursors; Humans; Lysosomes; Pancreatitis; Trypsinogen

Cystic fibrosis (CF), the most common lethal genetic disease affecting Caucasians, is a multi-system illness, most frequently characterized by childhood chronic obstructive pulmonary disease, pancreatic exocrine insufficiency, and abnormal sweat electrolyte concentrations. The diagnosis of CF is based on a combination of the above clinical findings and/or a positive family history of the illness in conjunction with an abnormal sweat test. The quantitative pilocarpine iontophoresis test is the sole acceptable method for diagnostic confirmation of the clinical suspicion of CF. A recent advance in the diagnosis of CF has been in the development of methods for neonatal detection. The immunoreactive trypsinogen (IRT) detection test is practical, adaptable to large scale screening of dried neonatal blood spots, relatively inexpensive, and promising for the detection of newborns with CF who have pancreatic insufficiency. However, the reliability and validity of this method have not yet been adequately established. Major advances in the treatment of patients with CF have emerged in the last decades, particularly in supportive pulmonary and nutritional care.

Topics: Adolescent; Anti-Bacterial Agents; Child; Child, Preschool; Chlorides; Cystic Fibrosis; Humans; Infant; Infant, Newborn; Mass Screening; Pancreatitis; Pilocarpine; Respiratory Tract Infections; Sweat; Trypsinogen

Topics: Animals; Clinical Enzyme Tests; Diagnostic Errors; Enzyme Precursors; Humans; Pancreas; Pancreatic Juice; Pancreatitis; Substrate Specificity; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Acute Disease; Clinical Enzyme Tests; Humans; Pancreas; Pancreatitis; Peptide Hydrolases; Trypsin; Trypsinogen

Topics: Acute Disease; Alcoholism; Calcinosis; Cholecystitis; Chronic Disease; Enzyme Activation; Humans; Kinins; Lactoferrin; Pancreatitis; Protein Biosynthesis; Protein-Energy Malnutrition; Shock; Trypsin; Trypsinogen

Topics: Acute Disease; Alcoholism; alpha 1-Antitrypsin; alpha-Macroglobulins; Cholelithiasis; Critical Care; Enzyme Activation; Humans; Metabolic Diseases; Pancreatitis; Parasympatholytics; Peritoneal Dialysis; Trypsinogen

Topics: Amylases; Aprotinin; Cathepsins; Kallikreins; Kinins; Lipase; Models, Theoretical; Necrosis; Pancreas; Pancreatic Elastase; Pancreatic Juice; Pancreatitis; Peptide Hydrolases; Trypsin; Trypsinogen

Ulinastatin, an intrinsic trypsin inhibitor, has proved to be effective for the prevention of acute pancreatitis after endoscopic retrograde cholangiopancreatography. The aim of this study was to assess the efficacy of ulinastatin for postoperative pancreatitis following pancreaticoduodenectomy in a randomized clinical trial.. Patients undergoing pancreaticoduodenectomy were randomized to receive perioperative ulinastatin or placebo. Levels of serum amylase, drain amylase, and urine trypsinogen-2 were measured.. A total of 42 patients were enrolled (20 in the ulinastatin group, 20 in the placebo group, 2 excluded). Two patients in the ulinastatin group and nine patients in the placebo group developed hyperamylasemia (P = 0.013) No patient in the ulinastatin group and five patients in the placebo group developed pancreatitis (P = 0.016). One patient in the ulinastatin group and two patients in the placebo group developed grade A pancreatic fistula (P = 0.548). Serum amylase levels at 4 hr and postoperative days 1, 2, and 3, and drain amylase levels on days 2 and 3 were significantly lower in the ulinastatin group than in the placebo group.. Prophylactic administration of ulinastatin reduced the levels of serum and drain amylase and the incidence of postoperative pancreatitis following pancreaticoduodenectomy.

Topics: Adult; Aged; Aged, 80 and over; Amylases; Double-Blind Method; Female; Glycoproteins; Humans; Male; Middle Aged; Pancreaticoduodenectomy; Pancreatitis; Treatment Outcome; Trypsin; Trypsin Inhibitors; Trypsinogen

To evaluate the use of the trypsinogen-2 dipstick (Actim Pancreatitis) test for early diagnosis and prediction of severity in acute pancreatitis (AP).. Ninety-two patients with AP were included in this study. The control group was 25 patients who had acute abdominal pain from non-pancreatic causes. Urine trypsinogen-2 dipstick test (UTDT) and conventional diagnostic tests were performed in all patients. Patients were divided by the Atlanta classification into two groups as having mild or severe pancreatitis.. UTDT was positive in 87 (94.6%) of the AP patients and in two (8%) controls (P < 0.05). Positive UTDT was found in 61 (92.4%) of 66 (71.7%) patients with mild pancreatitis and in all (100%) of the 26 (28.3%) with severe pancreatitis (P > 0.05). UTDT positivity lasted longer in severe pancreatitis compared with that in mild pancreatitis (6.2 +/- 2.5 d vs 2.0 +/- 1.43 d, P < 0.05). The sensitivity, specificity, positive predictive value, negative predictive value (NPV), positive likelihood ratio (PLR) and negative likelihood ratio (NLR) of UTDT were 91%, 72%, 96.6%, 70.4%, 3.4 and 0.1, respectively.. UTDT is a simple, rapid and reliable method for use on admission. It has high specificity and low NLR for early diagnosis and prediction of severity in AP. However, its relatively low NPV does not allow trypsinogen-2 dipstick test to be a stand-alone tool for diagnosis of acute pancreatitis; the use of other conventional diagnostic tools remains a requirement.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; APACHE; Early Diagnosis; Female; Humans; Lipase; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Prognosis; Prospective Studies; Reagent Kits, Diagnostic; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen

Hereditary pancreatitis (HP) is a form of recurrent acute pancreatitis (AP) mediated by mutations in cationic trypsinogen (PRSS1). Mutations cluster in the calcium-associated regulator regions of PRSS1. In rats, calcium-channel blockers (CCB) prevent hyperstimulation-associated AP. Because of the potential importance of hyperstimulation in triggering episodes of AP in HP, we designed a pilot study to evaluate the safety and potential benefit of CCB use in HP.. Subjects 6 years or older had a PRSS1 mutation, recurrent AP, and pain. Total study duration was 16 weeks. Amlodipine was given during weeks 0 to 11. Dose (2.5, 5, or 10 mg) was based on weight (range, 0.08-0.17 mg x kg(-1) x d(-1)). Subjects filled a daily diary including pain (0-10 scale) and blood pressure reading. Clinical assessments occurred at weeks -4, 0, 1, 2, 6, 10, 11, and 12. Subjects filled a Medical Outcomes Study Short-Form Survey version 2 (SF-10 for children <14 years old) at weeks -4, 0, 6, and 10. Data were compared for weeks -4 to 0 and 6 to 10.. Nine subjects signed informed consent (4 males; 12-52 years old). Four were excluded during the screening phase. Drug was discontinued in one due to development of unilateral lower-extremity numbness. Four subjects (12-31 years old) completed the study. Mean blood pressure, laboratory tests, physical findings, and daily pain scores did not clinically significantly differ before and during drug therapy, but all reported reduced symptoms. Three reduced analgesic use. Three had improved scores on the Medical Outcomes Study Short-Form Survey version 2.. Amlodipine is generally safe in subjects with HP and does not increase pain or episodes of AP. Further research into the mechanism of CCB on pancreatitis would be important to provide a pathophysiologic basis to support further trials in HP.

Topics: Acute Disease; Adolescent; Adult; Amlodipine; Analgesics; Blood Pressure; Calcium Channel Blockers; Child; Feasibility Studies; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Mutation; Pain; Pain Measurement; Pancreatitis; Patient Dropouts; Pilot Projects; Prospective Studies; Quality of Life; Recurrence; Risk Factors; Severity of Illness Index; Surveys and Questionnaires; Time Factors; Treatment Outcome; Trypsin; Trypsinogen

Early diagnosis of acute pancreatitis remains a challenge. A rapid dipstick screening test for acute pancreatitis has been developed. This prospective study was designed to evaluate the diagnostic value and time course of the rapid urinary trypsinogen-2 test strip in acute pancreatitis, with comparisons with serum amylase and serum lipase.. A total of 165 patients with acute abdominal pain (67 with acute pancreatitis and 98 with other acute abdominal diseases) attending our emergency unit were included. All patients were tested with the urinary trypsinogen-2 test strip, and serum amylase and serum lipase concentrations were determined simultaneously. To measure the time course of the urinary trypsinogen-2 test, 32 patients with acute pancreatitis were tested with a urinary trypsinogen-2 test strip on days 1, 2, 3, and 4 after admission.. Using a cutoff level of 50 microg/L for urinary trypsinogen-2, the sensitivity, specificity, and accuracy of the urinary trypsinogen-2 test strip for recognition of acute pancreatitis were 89.6%, 85.7%, and 87.3%, respectively. The diagnostic accuracy rates of serum amylase and serum lipase were 88.5% and 93.3%, using cutoff values of 3 times the upper normal limits for serum amylase and serum lipase, respectively. All but one of the 17 patients with severe acute pancreatitis was detected by the test strip (sensitivity, 94.1%). The time-course study of the urinary trypsinogen-2 test strip revealed that the sensitivity on days 1, 2, 3, and 4 was 90.6%, 81.2%, 59.4%, and 50%, respectively. There was no significant difference in the sensitivity between urinary trypsinogen-2 and serum lipase; however, the sensitivity values of serum lipase were significantly higher than those of serum amylase from days 1 to 4.. The rapid urinary trypsinogen-2 test is a reliable and simple method for the early diagnosis of acute pancreatitis. A positive test identifies patients in need of further diagnostic measures. The urinary trypsinogen-2 test can be performed in health care units where laboratory testing facilities are not immediately available.

Topics: Abdominal Pain; Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Diagnosis, Differential; Female; Humans; Lipase; Male; Middle Aged; Pancreatitis; Reagent Strips; Reproducibility of Results; Sensitivity and Specificity; Time Factors; Trypsin; Trypsinogen

The CFTR gene has been shown to be involved in sporadic idiopathic pancreatitis (IP) and neonatal hypertrypsinemia with normal sweat chloride test (NHNST). The cationic trypsinogen gene (Try4) is responsible for hereditary pancreatitis. The aim of the present study was to find a correlation between mutations in the two genes and the two phenotypes.. Analysis of some known gene mutations and complete gene screening by denaturing gradient gel electrophoresis and DNA sequencing were undertaken. Thirty-two sporadic IP patients were investigated for the CFTR study, while 13 sporadic IP patients plus 4 hereditary pancreatitis families (24 tested individuals) were examined for the Try4 study. Fifty neonates with NHNST were investigated for the study of both genes.. CFTR mutations were more frequently observed in sporadic IP cases with a common cystic fibrosis mutation or borderline sweat chloride than in cases with a negative sweat test. Try4 mutations were found in 1 out of the 13 sporadic IP cases tested.. The CFTR gene may be involved in IP and NHNST, while the Try4 gene may be involved in IP, but not in NHNST, in this limited series of observations.

Topics: Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Humans; Infant, Newborn; Infant, Newborn, Diseases; Mutation; Neonatal Screening; Pancreatitis; Pedigree; Phenotype; Polymorphism, Genetic; Sweat; Trypsin; Trypsinogen

Topics: alpha 1-Antitrypsin; Biomarkers; Cholangiopancreatography, Endoscopic Retrograde; Humans; Pancreatitis; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity; Trypsin; Trypsin Inhibitors; Trypsinogen

We have evaluated a new urinary trypsinogen-2 test strip, based on the principle of immunochromatography, in the diagnosis of acute pancreatitis induced by endoscopic retrograde cholangiopancreatography (ERCP).. One hundred six consecutive patients undergoing ERCP (with opacification of the pancreatic duct) at the Helsinki University Central Hospital were included in the study. Patients were tested with a urinary trypsinogen-2 test strip six hours after ERCP. Quantitative trypsinogen-2 as well as serum and urine amylase values were measured before the procedure and six hours after it.. In patients developing pancreatitis after ERCP, the median urinary trypsinogen-2 concentration six hours after the endoscopic procedure was 1780 micrograms/l (range 29-10,700 micrograms/l), and in patients without pancreatitis the median concentration was 3.6 micrograms/l (range 0.1-3390 micrograms/l; P < 0.0001). The sensitivity and specificity figures for the urinary trypsinogen-2 test strip results in diagnosing post-ERCP pancreatitis were comparable (81% and 97%, respectively) to those for serum amylase (91% and 96%) and urine amylase measurements (81% and 95%). The test strip showed a good correlation (kappa = 0.75) with the quantitative trypsinogen-2 assay.. The increase in urinary trypsinogen-2 concentration after ERCP reflects pancreatic injury, and can be detected by the test strip. Patients should be tested before the ERCP procedure as well, since elevated baseline values occur. The test is reliable and easy to perform even on an outpatient basis. However, its clinical usefulness requires evaluation in further trials.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Cholangiopancreatography, Endoscopic Retrograde; Diagnosis, Differential; Female; Humans; Male; Middle Aged; Pancreatitis; Reagent Strips; Sensitivity and Specificity; Trypsinogen; Urinalysis

Topics: Abdominal Pain; Acute Disease; Diagnosis, Differential; Humans; Mass Screening; Pancreatitis; Prospective Studies; Sensitivity and Specificity; Trypsinogen

A simple, rapid test is specific and sensitive enough to distinguish, in patients with clinically suspected acute pancreatitis, those whose abdominal pain is indeed of pancreatic origin has proved elusive.. In two consecutive series of surgical patients in a teaching hospital, whose acute abdominal pain turned out to be due to acute pancreatitis (n-57) or extrapancreatic in origin (n=40), we studied urinary trypsinogen-2 in two ways. A test strip, incorporating monoclonal antibodies to two epitopes on trypsinogen-2, recorded a blue line when concentrations exceeded 50 microgram/L; we also measured trypsinogen-2 concentrations in the laboratory.. In the patients with acute pancreatitis the test strip was positive in 52 and negative in five, whereas in the 40 extrapancreatic controls there were four false positives. In a further set of 57 orthopaedic controls, one urine was strip-test positive. Concentrations of urinary trypsinogen-2 and the test-strip results were in good agreement and in only three of the 154 patients were the two approaches discrepant, at the 50 microgram/L cut-off.. These findings, in patients whose acute abdominal pain was known to be pancreatic in origin or not, are encouraging but need to be confirmed in a consecutive series of patients in whom the diagnosis of pancreatitis is in doubt.

Topics: Abdominal Pain; Acute Disease; Adult; Aged; Aged, 80 and over; Diagnosis, Differential; False Positive Reactions; Female; Humans; Male; Middle Aged; Pancreatitis; Reagent Strips; Trypsin; Trypsinogen

To assess the relation between circulating cathodic trypsinogen (CT) levels and exocrine pancreatic function, and to compare the radioimmunological with the enzymatic measurement of duodenal trypsin, we evaluated exocrine pancreatic function in 34 controls and in 32 patients with proven chronic pancreatitis (CP). There was no relation between CT and the volume rate of pancreatic secretion, nor did serum CT levels correlate with the concentration output of duodenal bicarbonate in controls. However, in CP patients, there was a low value of the correlation coefficient. A good relationship between serum CT levels and duodenal trypsin secretion was detected when the trypsin content was expressed as the mean value of both concentration and output. The enzymatic estimation of duodenal trypsin was related closely to its radioimmunological measurement, but there was better correlation of serum CT to duodenal immunoreactive than to enzymatic trypsin. In patients with CP, low levels were observed in 29% of cases with serum CT estimation, in 75% with duodenal bicarbonate, and in 63% and 79% with enzymatic and immunoreactive trypsin outputs, respectively. We conclude that serum CT levels may reflect the functioning mass of pancreatic acinar cells and that in assessing pancreatic secretory capacity, the immunoreactive measurement of trypsin excretion is more sensitive than the enzymatic measurement and as accurate as bicarbonate output.

Topics: Bicarbonates; Chronic Disease; Clinical Enzyme Tests; Female; Humans; Male; Pancreas; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Topics: Animals; Cathepsin B; Ceruletide; Mice; Pancreas; Pancreatitis; Secretagogues; Trypsin; Trypsinogen

Topics: Animals; Mice; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

Mutation p.R122H in human cationic trypsinogen (PRSS1) is the most frequently identified cause of hereditary pancreatitis. The mutation blocks protective degradation of trypsinogen by chymotrypsin C (CTRC), which involves an obligatory trypsin-mediated cleavage at Arg122. Previously, we found that C57BL/6N mice are naturally deficient in CTRC, and trypsinogen degradation is catalyzed by chymotrypsin B1 (CTRB1). Here, we used biochemical experiments to demonstrate that the cognate p.R123H mutation in mouse cationic trypsinogen (isoform T7) only partially prevented CTRB1-mediated degradation. We generated a novel C57BL/6N mouse strain harboring the p.R123H mutation in the native T7 trypsinogen locus. T7R123H mice developed no spontaneous pancreatitis, and severity parameters of cerulein-induced pancreatitis trended only slightly higher than those of C57BL/6N mice. However, when treated with cerulein for 2 days, more edema and higher trypsin activity was seen in the pancreas of T7R123H mice compared to C57BL/6N controls. Furthermore, about 40% of T7R123H mice progressed to atrophic pancreatitis in 3 days, whereas C57BL/6N animals showed full histological recovery. Taken together, the observations indicate that mutation p.R123H inefficiently blocks chymotrypsin-mediated degradation of mouse cationic trypsinogen, and modestly increases cerulein-induced intrapancreatic trypsin activity and pancreatitis severity. The findings support the notion that the pathogenic effect of the PRSS1 p.R122H mutation in hereditary pancreatitis is dependent on its ability to defuse chymotrypsin-dependent defenses.

Topics: Animals; Ceruletide; Chymotrypsin; Humans; Mice; Mice, Inbred C57BL; Mutation; Pancreatitis; Trypsin; Trypsinogen

Chymotrypsin-like protease (CTRL) is one of the four chymotrypsin isoforms expressed in the human exocrine pancreas. Human genetic and experimental evidence indicate that chymotrypsins B1, B2, and C (CTRB1, CTRB2 and CTRC) are important not only for protein digestion but also for protecting the pancreas against pancreatitis by degrading potentially harmful trypsinogen. CTRL has not been reported to play a similar role, possibly due to its low abundance and/or different substrate specificity. To address this problem, we investigated the specificity of the substrate-binding groove of CTRL by evolving the substrate-like canonical loop of the Schistocerca gregaria proteinase inhibitor 2 (SGPI-2), a small-protein reversible chymotrypsin inhibitor to bind CTRL. We found that phage-associated SGPI-2 variants with strong affinity to CTRL were similar to those evolved previously against CTRB1, CTRB2 or bovine chymotrypsin A (bCTRA), indicating comparable substrate specificity. When tested as recombinant proteins, SGPI-2 variants inhibited CTRL with similar or slightly weaker affinity than bCTRA, confirming that CTRL is a typical chymotrypsin. Interestingly, an SGPI-2 variant selected with a Thr29His mutation in its reactive loop was found to inhibit CTRL strongly, but it was digested rapidly by bCTRA. Finally, CTRL was shown to degrade human anionic trypsinogen, however, at a much slower rate than CTRB2, suggesting that CTRL may not have a significant role in the pancreatic defense mechanisms against inappropriate trypsinogen activation and pancreatitis.

Topics: Animals; Cattle; Chymases; Chymotrypsin; Humans; Pancreatitis; Peptide Library; Protease Inhibitors; Substrate Specificity; Trypsinogen

Acute pancreatitis (AP) is a common gastrointestinal disease with substantial morbidity and mortality. Pancreatic acinar intracellular trypsinogen activation (PAITA) is an important event in the early stage of AP. The present study aimed to investigate the effects of tRNA-derived fragments (tRFs) and the microRNA regulatory network on pancreatic acinar intracellular trypsinogen activation (PAITA) and identify novel key targets in AP. Taurolithocholic acid 3-sulfate (TLC-S)-treated AR42J cells were used to establish a PAITA model. Twenty differentially expressed tRFs and 35 DE microRNAs were identified in PAITA through gene sequencing. Based on these genes, we established the tRF-mRNA and microRNA-mRNA regulatory networks by using bioinformatics methods. The networks revealed 29 hub microRNAs (e.g., Let-7 family, miR-21-3p.) and 19 hub tRFs (e.g., tRF3-Thr-AGT, i-tRF-Met-CAT) in PAITA. GO analysis showed that the functions of the two networks were similar and mainly enriched in RNA splicing, mRNA processing, and so on. tRF3-Thr-AGT, targeting Btg2, Cd44, Zbp1, etc., was significantly decreased in PAITA. Moreover, the trypsinogen activation level was increased significantly in the tRF3-Thr-AGT deficiency groups, but rescued by tRF3-Thr-AGT mimics. The results revealed that downregulated tRF3-Thr-AGT was involved in PAITA. This study provides potential novel targets for researching the underlying mechanisms of AP.

Topics: Acinar Cells; Animals; Cell Line, Tumor; Enzyme Activation; MicroRNAs; Pancreatitis; Rats; RNA, Transfer; Trypsinogen

T7K24R mice carry mutation p.K24R in mouse cationic trypsinogen (isoform T7), which is analogous to the human hereditary pancreatitis-associated mutation p.K23R. The mutation renders trypsinogen more prone to autoactivation. We recently reported that T7K24R mice exhibit increased severity of acute pancreatitis induced by repeated cerulein injections. The objective of the present study was to test whether trypsinogen mutant mice are prone to develop chronic pancreatitis, as observed in patients. We characterized the natural course of cerulein-induced pancreatitis in T7K24R mice and the C57BL/6N parent strain from the acute episode to 3 months post-attack. As expected, an acute episode of pancreatitis in C57BL/6N mice was followed by rapid recovery and histological restitution. In stark contrast, T7K24R mice developed progressive chronic pancreatitis with acinar cell atrophy, persistent macrophage infiltration, and diffuse fibrosis. The nadir of pancreas damage occurred on days 5-6 after the acute episode and was accompanied by digestive dysfunction. Remarkably, histological recovery was markedly delayed and permanent, chronic changes were still detectable 1-3 months after the acute pancreatitis episode. We conclude that during cerulein-induced acute pancreatitis in T7K24R mice, trypsin triggers an autonomous inflammatory program resulting in chronic disease progression, even after the cessation of cerulein-mediated injury. We propose that this uniquely trypsin-dependent mechanism explains the development of hereditary chronic pancreatitis in humans. Trypsin inhibition during acute attacks should prevent or delay progression to chronic disease.

Topics: Acute Disease; Animals; Ceruletide; Humans; Mice; Mice, Inbred C57BL; Pancreas; Pancreatitis; Trypsinogen

Topics: Animals; Chymotrypsin; Mice; Pancreatitis; Phenotype; Trypsin; Trypsinogen

Pancreatic chymotrypsins (CTRs) are digestive proteases that in humans include CTRB1, CTRB2, CTRC, and CTRL. The highly similar CTRB1 and CTRB2 are the products of gene duplication. A common inversion at the CTRB1-CTRB2 locus reverses the expression ratio of these isoforms in favor of CTRB2. Carriers of the inversion allele are protected against the inflammatory disorder pancreatitis presumably via their increased capacity for CTRB2-mediated degradation of harmful trypsinogen. To reveal the protective molecular determinants of CTRB2, we compared enzymatic properties of CTRB1, CTRB2, and bovine CTRA (bCTRA). By evolving substrate-like Schistocerca gregaria proteinase inhibitor 2 (SGPI-2) inhibitory loop variants against the chymotrypsins, we found that the substrate binding groove of the three enzymes had overlapping specificities. Based on the selected sequences, we produced eight SGPI-2 variants. Remarkably, CTRB2 and bCTRA bound these inhibitors with significantly higher affinity than CTRB1. Moreover, digestion of peptide substrates, beta casein, and human anionic trypsinogen unequivocally confirmed that CTRB2 is a generally better enzyme than CTRB1 while the potency of bCTRA lies between those of the human isoforms. Unexpectedly, mutation D236R alone converted CTRB1 to a CTRB2-like high activity protease. Modeling indicated that in CTRB1 Met210 partially obstructed the substrate binding groove, which was relieved by the D236R mutation. Taken together, we identify CTRB2 Arg236 as a key positive determinant, while CTRB1 Asp236 as a negative determinant for chymotrypsin activity. These findings strongly support the concept that in carriers of the CTRB1-CTRB2 inversion allele, the superior trypsinogen degradation capacity of CTRB2 protects against pancreatitis.

Topics: Animals; Cattle; Chymotrypsin; Humans; Pancreas; Pancreatitis; Peptides; Trypsinogen

Premature intracellular trypsinogen activation has long been considered a key initiator of acute pancreatitis (AP). Cathepsin B (CTSB) activates trypsinogen, while cathepsin L (CTSL) inactivates trypsin(ogen), and both proteins play a role in the onset of AP.. AP was induced by 7 hourly intraperitoneal injections of cerulein (50 μg/kg) in wild-type and pancreas-specific conditional Ctsb knockout (Ctsb. Double deletion of Ctsb and Cstl did not affect pancreatic development or mouse growth. After 7 times cerulein injections, double Ctsb and Ctsl deficiency in mouse pancreases increased trypsin activity to the same extent as that in Ctsl-deficient mice, while Ctsb deficiency decreased trypsin activity but did not affect the severity of AP. Ctsb. Double deletion of Ctsb and Ctsl in the mouse pancreas altered intrapancreatic trypsin activity but did not affect disease severity and inflammatory response after cerulein-induced AP.

Topics: Acute Disease; Amylases; Animals; Cathepsin B; Ceruletide; Mice; Mice, Knockout; Pancreas; Pancreatitis; Trypsin; Trypsinogen

Acute pancreatitis is the sudden inflammation of the pancreas. Severe cases of acute pancreatitis are potentially fatal and have no specific treatment available. Premature trypsinogen activation could initiate acute pancreatitis. However, the mechanism underlying premature trypsinogen activation is not fully understood.. In this research, a primary pancreatic acinar cell or mouse acute pancreatitis model was constructed. The effect of acid ceramidase (ASAH1), which is responsible for sphingosine production, was investigated in trypsinogen activation in vitro and in vivo. Meanwhile, the proteins regulating ASAH1 or binding to sphingosine were also detected by co-immunoprecipitation followed by mass spectrometry.. The results showed that ASAH1 increased in acute pancreatitis. Increased ASAH1 promoted the activation of trypsinogen and cathepsin B. On the contrary, ASAH1 downregulation inhibited trypsinogen and cathepsin B. Meanwhile, ASAH1 regulated the activity of trypsin and cathepsin B through sphingosine. Additionally, E3 ligase Mind bomb homolog 1 (MIB1) decreased in acute pancreatitis resulting in the decreased binding between MIB1 and ASAH1. Exogenous MIB1 diminished the elevation in trypsin activity induced by acute pancreatitis inducer. ASAH1 increased owing to the inhibition of the proteasome degradation by MIB1. In acute pancreatitis, sphingosine was found to bind to pyruvate kinase. Pyruvate kinase activation could reduce trypsinogen activation and mitochondrial reactive oxygen species (ROS) production induced by sphingosine.. In conclusion, during the process of acute pancreatitis, MIB1 downregulation led to ASAH1 upregulation, resulting in pyruvate kinase inhibition, followed by trypsinogen activation.

Topics: Acid Ceramidase; Acute Disease; Animals; Cathepsin B; Disease Models, Animal; Mice; Pancreatitis; Pyruvate Kinase; Sphingosine; Trypsin; Trypsinogen

Pancreatitis, the inflammatory disorder of the pancreas, has no specific therapy. Genetic, biochemical, and animal model studies revealed that trypsin plays a central role in the onset and progression of pancreatitis. Here, we performed biochemical and preclinical mouse experiments to offer proof of concept that orally administered dabigatran etexilate can inhibit pancreatic trypsins and shows therapeutic efficacy in trypsin-dependent pancreatitis. We found that dabigatran competitively inhibited all human and mouse trypsin isoforms (Ki range 10-79 nM) and dabigatran plasma concentrations in mice given oral dabigatran etexilate well exceeded the Ki of trypsin inhibition. In the T7K24R trypsinogen mutant mouse model, a single oral gavage of dabigatran etexilate was effective against cerulein-induced progressive pancreatitis, with a high degree of histological normalization. In contrast, spontaneous pancreatitis in T7D23A mice, which carry a more aggressive trypsinogen mutation, was not ameliorated by dabigatran etexilate, given either as daily gavages or by mixing it with solid chow. Taken together, our observations showed that benzamidine derivatives such as dabigatran are potent trypsin inhibitors and show therapeutic activity against trypsin-dependent pancreatitis in T7K24R mice. Lack of efficacy in T7D23A mice is probably related to the more severe pathology and insufficient drug concentrations in the pancreas.

Topics: Animals; Dabigatran; Disease Models, Animal; Humans; Mice; Pancreas; Pancreatitis; Trypsin; Trypsinogen

In acute pancreatitis, activation of inflammatory signaling, including the nuclear factor-kappa B (NF-κB) pathway, within acinar cells is known to be an early intracellular event occurring in parallel with pathologic trypsinogen activation. Sphingosine 1-phosphate receptor 2 (S1PR2) plays a critical role in endothelial inflammation, and our previous studies reported that S1PR2 deficiency significantly reduced the inflammatory response in liver injury under cholestasis conditions. However, the role of S1PR2 in inflammatory signaling activation within acinar cells and inflammatory responses during acute pancreatitis has not been elucidated. Here we report that S1PR2 was upregulated in the whole pancreas during acute pancreatitis. Blockade of S1PR2 by pharmacologic inhibition of S1PR2 by JTE-013 or AAV-mediated knockdown of S1PR2 improved the severity of pancreatic injury, as indicated by a significant reduction in inflammation and acinar cells death in acute pancreatitis mice. Moreover, S1PR2 is the predominant S1PRs expressed in pancreatic acinar cells and mediates NF-κB activation and the early inflammatory response within acinar cells under acute pancreatitis conditions via ROCK signaling pathways, not extracellular signal-regulated kinase pathways or p38 mitogen-activated protein kinase pathways. In addition, S1PR2 mediated macrophage NF-κB activation, migration and polarization toward the M1 phenotype. Therefore, these results demonstrated that the S1PR2-mediated early inflammatory response in acinar cells promotes the progression of acute pancreatitis, successfully linking local events to the systematic inflammatory response and leading to a novel therapeutic target for acute pancreatitis aimed at halting the progression of the inflammatory response. Video Abstract.

Topics: Acute Disease; Animals; Inflammation; Mice; NF-kappa B; p38 Mitogen-Activated Protein Kinases; Pancreas; Pancreatitis; Sphingosine-1-Phosphate Receptors; Trypsinogen

Topics: Adult; Diagnosis, Differential; Emergency Service, Hospital; Female; Humans; Male; Middle Aged; Pancreatitis; Point-of-Care Systems; Point-of-Care Testing; Prospective Studies; Sensitivity and Specificity; Trypsinogen

A dipstick test for urine trypsinogen-2 has been used in the diagnosis of acute pancreatitis, but there are only a few studies exploring the effectiveness of this test for early diagnose of post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis (PEP).. The authors explore if the rapid point-of-care urine trypsinogen-2 dipstick test can replace assay of amylase in diagnosing PEP.. For this prospective study, from Helsinki University Hospital 400 ERCP patients were enrolled in whom the authors analyzed plasma amylase or pancreas-specific amylase, bilirubin, and urine trypsinogen-2, and urine trypsinogen-2 with dipstick before, 4 and 24 hours after ERCP.. PEP developed in 15 (3.8%) patients. Urine trypsinogen-2 concentrations were significantly higher in PEP than in non-PEP patients 24 hours after ERCP (P=0.001, Mann-Whitney U test) but not 4 hours after ERCP (P=0.094). When combined with abdominal pain symptoms at 4 hours the dipstick test had a sensitivity of 60%, a specificity of 99%, a positive predictive value of 64%, and a negative predictive value 98%. At 24 hours, sensitivity was 100%, specificity 98%, positive predictive value 71%, and negative predictive value 100%.. A positive dipstick seems to identify PEP cases and a negative test excludes PEP with high accuracy.

Topics: Acute Disease; Cholangiopancreatography, Endoscopic Retrograde; Humans; Pancreatitis; Prospective Studies; Trypsinogen

Mesotrypsin is a low-abundance human trypsin isoform with a unique evolutionary mutation that conferred resistance to trypsin inhibitors and restricted substrate specificity. Mesotrypsin degrades the serine protease inhibitor Kazal type 1 (SPINK1) and thereby might increase risk for pancreatitis. Here, we report a mouse model designed to test the role of mesotrypsin in pancreatitis. We introduced the human mesotrypsin evolutionary signature mutation into mouse cationic trypsinogen (isoform T7), resulting in a Gly to Arg change at the corresponding position 199. In biochemical experiments using purified proteins, the p.G199R T7 mutant recapitulated all salient features of human mesotrypsin. T7G199R mice developed normally with no spontaneous pancreatitis or other obvious phenotypic changes. Cerulein-induced acute pancreatitis in C57BL/6N and T7G199R mice showed similar severity with respect to inflammatory parameters and acinar cell necrosis while plasma amylase activity was higher in T7G199R mice. Neither SPINK1 degradation nor elevated intrapancreatic trypsin activation was apparent in T7G199R mice. The results indicate that in T7G199R mice the newly created mesotrypsin-like activity has no significant impact on cerulein-induced pancreatitis. The observations suggest that human mesotrypsin is unimportant for pancreatitis; a notion that is consistent with published human genetic studies.

Topics: Animals; Ceruletide; Chymotrypsin; Disease Models, Animal; Gene Expression Regulation; Glycoproteins; Humans; Inflammation; Mice; Mice, Inbred C57BL; Mutation; Pancreatitis; Prostatic Secretory Proteins; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Mutations in the trypsinogen gene (PRSS1) cause human hereditary pancreatitis. However, it is not clear how mutant forms of PRSS1 contribute to disease development. We studied the effects of expressing mutant forms of human PRSS1 in mice.. We expressed forms of PRSS1 with and without the mutation encoding R122H (PRSS1. Pancreata from mice expressing transgenes encoding PRSS1 or PRSS1. Expression of a transgene encoding PRSS1

Topics: Acinar Cells; Adaptive Immunity; Animals; Gene Expression; Humans; Mice; Mice, Transgenic; Mutation; Pancreas; Pancreatitis; Transgenes; Trypsin; Trypsinogen

Currently, an effective targeted therapy for pancreatitis is lacking. Hereditary pancreatitis (HP) is a heritable, autosomal-dominant disorder with recurrent acute pancreatitis (AP) progressing to chronic pancreatitis (CP) and a markedly increased risk of pancreatic cancer. In 1996, mutations in PRSS1 were linked to the development of HP. Here, we developed a mouse model by inserting a full-length human PRSS1R122H gene, the most commonly mutated gene in human HP, into mice. Expression of PRSS1R122H protein in the pancreas markedly increased stress signaling pathways and exacerbated AP. After the attack of AP, all PRSS1R122H mice had disease progression to CP, with similar histologic features as those observed in human HP. By comparing PRSS1R122H mice with PRSS1WT mice, as well as enzymatically inactivated Dead-PRSS1R122H mice, we unraveled that increased trypsin activity is the mechanism for R122H mutation to sensitize mice to the development of pancreatitis. We further discovered that trypsin inhibition, in combination with anticoagulation therapy, synergistically prevented progression to CP in PRSS1R122H mice. These animal models help us better understand the complex nature of this disease and provide powerful tools for developing and testing novel therapeutics for human pancreatitis.

Topics: Animals; Anticoagulants; Disease Models, Animal; Disease Susceptibility; Humans; Mice; Mice, Transgenic; Mutation; Pancreas; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

Mutations in the human serine protease 1 gene (PRSS1), which encodes cationic trypsinogen, can accelerate its autoactivation and cause hereditary or sporadic chronic pancreatitis. Disruption of the locus that encodes cationic trypsinogen in mice (T7) causes loss of expression of the protein, but only partially decreases the severity of secretagogue-induced acute pancreatitis and has no effect on chronic pancreatitis. We investigated whether trypsinogen becomes pathogenic only when its activation is promoted by mutation.. We generated mice with knock-in of the p.K24R mutation (called T7K24R mice), which is analogous to human PRSS1 mutation p.K23R. We gave T7K24R and C57BL/6N (control) mice repeated injections of cerulein to induce pancreatitis. Plasma amylase activity, pancreatic edema, and myeloperoxidase content in pancreas and lungs were quantified. We expressed mutant and full-length forms of PRSS1 in Escherichia coli and compared their autoactivation.. The p.K24R mutation increased autoactivation of T7 5-fold. T7K24R mice developed no spontaneous pancreatitis. T7K24R mice given cerulein injections had increased pancreatic activation of trypsinogen and more edema, infiltration of lung and pancreas by inflammatory cells, and plasma amylase activity compared with control mice given cerulein injections. Injection of cerulein for 2 days induced progressive pancreatitis in T7K24R mice, but not in control mice, with typical features of chronic pancreatitis.. Introduction of a mutation into mice that is analogous to the p.K23R mutation in PRSS1 increases pancreatic activation of trypsinogen during secretagogue-induced pancreatitis. Higher pancreatic activity of trypsin increases the severity of pancreatitis, even though loss of trypsin activity does not prevent pancreatitis in mice.

Topics: Animals; Mice; Mice, Inbred C57BL; Mutation; Pancreas; Pancreatitis; Pancreatitis, Chronic; Secretagogues; Severity of Illness Index; Trypsin; Trypsinogen

Topics: Animals; Mice; Pancreatitis; Trypsin; Trypsinogen

To study the role of kinase inhibitor PD98059 on autophagy flow in the process of trypsinogen activation in pancreatic acinar cell and its related mechanism.. In the present study, bioinformatics analysis was used to predict kinases and their most relevant inhibitor (PD98059) which participates in autophagy of acute pancreatitis (AP). The rat pancreatic acini AR42J cells were divided into 4 groups: control group, sodium taurocholate hydrate (TLC) group, PD98059 group, and TLC + PD group. Twenty-seven Sprague-Dawley rats were divided into 3 groups (n = 9), including control group, severe AP (SAP) group, and SAP + PD group. We detected trypsinogen activation, autophagic activation, lysosome pH, and cathepsin-L activity in vivo and in vitro.. Results revealed trypsinogen activation was significantly inhibited in mitogen-activated protein kinase 1, JAK2, LYN, and their common inhibitor was PD98059. The trypsinogen activation, Beclin1, and light chain 3 II expressions were reduced, whereas the expressions of lysosomal-associated membrane protein 2, cathepsin L1, and cathepsin-L activity is upregulated after the PD98059 pretreatment, both in vivo and in vitro.. Lysosomal dysfunction blocked autophagy flux, accompanied by increasing pancreatic acinar cell autophagy in the process of trypsinogen activation. PD98059 inhibited AP occurrence and pancreatic injury via improving the blocked autophagic pathway and reducing trypsinogen activation.

Topics: Acinar Cells; Acute Disease; Animals; Autophagosomes; Autophagy; Cell Line; Enzyme Activation; Flavonoids; Hydrogen-Ion Concentration; Lysosomes; Male; Pancreas; Pancreatitis; Protein Kinase Inhibitors; Rats, Sprague-Dawley; Trypsinogen

Trypsinogen activation is the hallmark of acute pancreatitis (AP) independent of intra-acinar NF-κB activation and inflammation. We previously found that dopamine (DA) receptor 2 (DRD2) activation controls inflammation during AP via PP2A-dependent NF-κB activation. In this study, we sought to examine whether DRD2 signaling mediates trypsinogen activation and the underlying mechanisms. Pancreatic acinar cells were stimulated with cholecystokinin-8 in vitro. AP was induced by intraperitoneal injections of caerulein and LPS or l-arginine. Pancreatitis severity was assessed biochemically and histologically. We found that activation of DRD2 by quinpirole, a potent DRD2 agonist, resulted in the reduction of trypsinogen activation and the upregulation of HSP70 in vitro and in vivo. Mechanistically, we found that quinpirole induced dephosphorylation of heat shock factor 1 (HSF1), a master transcription factor of HSP70, leading to increased nuclear translocation of HSF1 in a PP2A-dependent pathway. Furthermore, DRD2 activation restored lysosomal pH and, therefore, maintained lysosomal cathepsin B activity in a HSP70-dependent manner. VER155008, a potent HSP70 antagonist, abolished the protective effects observed with DRD2 activation in vitro and in two experimental models of AP. Our data showed that besides controlling NF-κB activation, DRD2 activation prevented trypsinogen activation during acute pancreatitis via PP2A-dependent upregulation of HSP70 and further support that DRD2 agonist could be a promising therapeutic strategy for treating AP.

Topics: Animals; Ceruletide; Dopamine Agonists; Gene Expression Regulation; Heat Shock Transcription Factors; HSP72 Heat-Shock Proteins; Lysosomes; Mice; Mice, Inbred C57BL; Pancreas; Pancreatitis; Phosphorylation; Protein Phosphatase 2; Quinpirole; Receptors, Dopamine D2; Trypsinogen; Up-Regulation

Acute pancreatitis (AP) is a common digestive disorder with high morbidity and mortality. The present study aimed to investigate the expression of early growth response protein 1 (Egr1), and the effect of competing endogenous (ce)RNA network on trypsinogen activation. Pancreatic acinar intracellular trypsinogen activation (PAITA) is an important event in the early stage of AP; however, the underlying mechanisms remain unclear. The present study used taurolithocholic acid 3‑sulfate (TLC‑S)‑treated AR42J cells (pancreatic cell line) to establish a PAITA model. A gene microarray and bioinformatics analysis was performed to identify the potential key targets in PAITA. The results demonstrated that Egr1, an important transcription factor, was significantly overexpressed in PAITA. In Egr1 small interfering (si)RNA‑transfected cells, Egr1 expression was decreased and trypsinogen activation was significantly decreased compared with negative control siRNA‑transfected cells, indicating that in TLC‑S‑induced PAITA, overexpression of Egr1 enhanced trypsinogen activation. A ceRNA network [mRNA‑microRNA (miRNA/miR)‑long non‑coding (lnc)RNA] generated using the PAITA model revealed that the effects of Egr1 on PAITA may be regulated by multiple ceRNA pairs, and the lncRNAs (including NONRATT022624 and NONRATT031002) and miRNAs [including Rattus norvegicus (rno)‑miR‑214‑3p and rno‑miR‑764‑5p] included in the ceRNA pairs may serve roles in PAITA by regulating the expression of Egr1. The results of the present study may provide novel targets for researching the underlying mechanisms of, and developing treatments for AP.

Topics: Animals; Cell Line; Computational Biology; Early Growth Response Protein 1; Enzyme Activation; Gene Expression Profiling; Gene Regulatory Networks; MicroRNAs; Models, Biological; Oligonucleotide Array Sequence Analysis; Pancreatitis; Rats; RNA, Long Noncoding; RNA, Small Interfering; Taurolithocholic Acid; Trypsinogen; Up-Regulation

Premature intrapancreatic trypsinogen activation is widely regarded as an initiating event for acute pancreatitis. Previous studies have alternatively implicated secretory vesicles, endosomes, lysosomes, or autophagosomes/autophagolysosomes as the primary site of trypsinogen activation, from which a cell-damaging proteolytic cascade originates. To identify the subcellular compartment of initial trypsinogen activation we performed a time-resolution analysis of the first 12 h of caerulein-induced pancreatitis in transgenic light chain 3 (LC3)-GFP autophagy reporter mice. Intrapancreatic trypsin activity increased within 60 min and serum amylase within 2 h, but fluorescent autophagosome formation only by 4 h of pancreatitis in parallel with a shift from cytosolic LC3-I to membranous LC3-II on Western blots. At 60 min, activated trypsin in heavier subcellular fractions was co-distributed with cathepsin B, but not with the autophagy markers LC3 or autophagy protein 16 (ATG16). Supramaximal caerulein stimulation of primary pancreatic acini derived from LC3-GFP mice revealed that trypsinogen activation is independent of autophagolysosome formation already during the first 15 min of exposure to caerulein. Co-localization studies (with GFP-LC3 autophagosomes versus Ile-Pro-Arg-AMC trypsin activity and immunogold-labelling of lysosomal-associated membrane protein 2 [LAMP-2] versus trypsinogen activation peptide [TAP]) indicated active trypsin in autophagolysosomes only at the later timepoints. In conclusion, during the initiating phase of caerulein-induced pancreatitis, premature protease activation develops independently of autophagolysosome formation and in vesicles arising from the secretory pathway. However, autophagy is likely to regulate overall intracellular trypsin activity during the later stages of this disease.

Topics: Animals; Autophagosomes; Autophagy; Ceruletide; Endosomes; Male; Mice; Mice, Inbred C57BL; Pancreatitis; Secretory Vesicles; Trypsin; Trypsinogen

The objective of this study was to determine if infants carrying 1 cystic fibrosis transmembrane receptor (CFTR) mutation demonstrate pancreatic inflammation in response to tobacco exposure.. Cystic fibrosis carrier infants aged 4 to 16 weeks were prospectively enrolled. Tobacco exposure was assessed by survey and maternal hair nicotine analysis. Serum immunoreactive trypsinogen (IRT) levels at birth and at the time of recruitment were analyzed relative to the presence or absence of tobacco exposure. The effect of the severity of the CFTR mutation carried by the infant on the tobacco-IRT relationship was also analyzed.. Forty-eight infants completed the study. Newborn screen and follow-up IRT levels were not different between exposed infants (19 by hair analysis) and nonexposed infants (29 by hair analysis). Follow-up IRT levels were lower in infants with more severe CFTR mutations (P = 0.005). There was no difference in follow-up IRT based on CFTR mutation severity in exposed infants. Nonexposed infants with milder CFTR mutations had higher median IRT values on follow-up testing than those with more severe CFTR mutations (P < 0.05).. The pancreas of cystic fibrosis carrier infants is affected by tobacco exposure, and those carrying less severe CFTR mutations may be more susceptible to tobacco effects.

Topics: Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Genetic Testing; Hair; Heterozygote; Humans; Infant; Infant, Newborn; Male; Mutation; Neonatal Screening; Nicotine; Pancreatitis; Pilot Projects; Pregnancy; Prenatal Exposure Delayed Effects; Prospective Studies; Smoking; Tobacco Smoke Pollution; Trypsinogen

Acute pancreatitis is characterized by an early intracellular protease activation and invasion of leukocytes into the pancreas. Cathepsins constitute a large group of lysosomal enzymes, that have been shown to modulate trypsinogen activation and neutrophil infiltration. Cathepsin G (CTSG) is a neutrophil serine protease of the chymotrypsin C family known to degrade extracellular matrix components and to have regulatory functions in inflammatory disorders. The aim of this study was to investigate the role of CTSG in pancreatitis. Isolated acinar cells were exposed to recombinant CTSG and supramaximal cholezystokinin stimulation. In CTSG

Topics: Acinar Cells; Animals; Cathepsin G; Cells, Cultured; Ceruletide; Disease Models, Animal; Gene Knockout Techniques; Granulocytes; Male; Mice; Neutrophil Infiltration; Neutrophils; Pancreatitis; Trypsinogen

Asparaginase-associated pancreatitis is a life-threatening toxicity to childhood acute lymphoblastic leukemia treatment. To elucidate genetic predisposition and asparaginase-associated pancreatitis pathogenesis, ten trial groups contributed remission samples from patients aged 1.0-17.9 years treated for acute lymphoblastic leukemia between 2000 and 2016. Cases (n=244) were defined by the presence of at least two of the following criteria: (i) abdominal pain; (ii) levels of pancreatic enzymes ≥3 × upper normal limit; and (iii) imaging compatible with pancreatitis. Controls (n=1320) completed intended asparaginase therapy, with 78% receiving ≥8 injections of pegylated-asparaginase, without developing asparaginase-associated pancreatitis. rs62228256 on 20q13.2 showed the strongest association with the development of asparaginase-associated pancreatitis (odds ratio=3.75;

Topics: Adolescent; Alleles; Antineoplastic Agents; Asparaginase; Child; Child, Preschool; Female; Genetic Association Studies; Genetic Predisposition to Disease; Genetic Variation; Genotype; Humans; Infant; Male; Models, Biological; Pancreatitis; Phenotype; Polyethylene Glycols; Polymorphism, Single Nucleotide; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Trypsin; Trypsinogen

Rapid urinary trypsinogen-2 dipstick test and levels of urinary trypsinogen-2 and trypsinogen activation peptide (TAP) concentration have been reported as prognostic markers for the diagnosis of acute pancreatitis.. To reconfirm the validity of all these markers in the diagnosis of acute pancreatitis by undertaking a multi-center study in Japan.. Patients with acute abdominal pain were recruited from 17 medical institutions in Japan from April 2009 to December 2012. Urinary and serum samples were collected twice, at enrollment and on the following day for measuring target markers. The diagnosis and severity assessment of acute pancreatitis were assessed based on prognostic factors and computed tomography (CT) Grade of the Japanese Ministry of Health, Labour, and Welfare criteria.. A total of 94 patients were enrolled during the study period. The trypsinogen-2 dipstick test was positive in 57 of 78 patients with acute pancreatitis (sensitivity, 73.1%) and in 6 of 16 patients with abdominal pain but without any evidence of acute pancreatitis (specificity, 62.5%). The area under the curve (AUC) score of urinary trypsinogen-2 according to prognostic factors was 0.704, which was highest in all parameter. The AUC scores of urinary trypsinogen-2 and TAP according to CT Grade were 0.701 and 0.692, respectively, which shows higher than other pancreatic enzymes. The levels of urinary trypsinogen-2 and TAP were significantly higher in patients with extended extra-pancreatic inflammation as evaluated by CT Grade.. We reconfirmed urinary trypsinogen-2 dipstick test is useful as a marker for the diagnosis of acute pancreatitis. Urinary trypsinogen-2 and TAP may be considered as useful markers to determine extra-pancreatic inflammation in acute pancreatitis.

Topics: Adult; Aged; Aged, 80 and over; Area Under Curve; Biomarkers; Female; Humans; Japan; Male; Middle Aged; Oligopeptides; Pancreatitis; Prognosis; Prospective Studies; Retrospective Studies; Severity of Illness Index; Trypsin; Trypsinogen

We explored prediction of severe acute pancreatitis (AP) and development of organ dysfunction (OD).. Serum concentrations of serine peptidase inhibitor Kazal type 1 (SPINK1), trypsinogen 1, trypsinogen 2, and trypsinogen 3, complex between trypsin 2 and α1-antitrypsin, serum C-reactive protein, creatinine, and pancreatic amylase were measured in 239 AP patients with disease onset within 72 hours.. SPINK1 distinguished most accurately patients who later developed severe AP. The area under the receiver operating characteristic curve for SPINK1 was 0.742, followed by trypsinogen 2 (0.726), complex between trypsin 2 and α1-antitrypsin (0.657), creatinine (0.656), trypsinogen 1 (0.652), trypsinogen 3 (0.557), and C-reactive protein (0.499). With a cutoff of 166 μg/L, SPINK1 had a specificity of 93%, a sensitivity of 48%, and diagnostic odds ratio of 11.52. In multivariate logistic regression analysis, only SPINK1 was an independent predictor of severe AP among patients presenting without OD on admission (P < 0.001).. Plasma levels of the biomarkers and creatinine correlated with the severity of AP and development of OD. In patients presenting without OD at admission, SPINK1 was an independent marker for later development of severe AP.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin; Biomarkers; C-Reactive Protein; Creatinine; Female; Humans; Male; Middle Aged; Pancreatitis; ROC Curve; Severity of Illness Index; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen; Young Adult

Intra-acinar trypsinogen activation occurs in the earliest stages of pancreatitis and is believed to play important roles in pancreatitis pathogenesis. However, the exact role of intra-acinar trypsin activity in pancreatitis remains elusive. Here, we aimed to examine the specific effects of intra-acinar trypsin activity on the development of pancreatitis using a transgenic mouse model. This transgenic mouse model allowed for the conditional expression of a mutant trypsinogen that can be activated specifically inside pancreatic acinar cells. We found that expression of this active mutated trypsin had no significant effect on triggering spontaneous pancreatitis. Instead, several protective compensatory mechanisms, including SPINK1 and heat shock proteins, were upregulated. Notably, these transgenic mice developed much more severe acute pancreatitis, compared with control mice, when challenged with caerulein. Elevated tissue edema, serum amylase, inflammatory cell infiltration and acinar cell apoptosis were dramatically associated with increased trypsin activity. Furthermore, chronic pathological changes were observed in the pancreas of all transgenic mice, including inflammatory cell infiltration, parenchymal atrophy and cell loss, fibrosis, and fatty replacement. These changes were not observed in control mice treated with caerulein. The alterations in pancreata from transgenic mice mimicked the histological changes common to human chronic pancreatitis. Taken together, we provided in vivo evidence that increased intra-acinar activation of trypsinogen plays an important role in the initiation and progression of both acute and chronic pancreatitis.

Topics: Acinar Cells; Animals; Disease Models, Animal; Mice; Pancreas, Exocrine; Pancreatitis; Pancreatitis, Chronic; Severity of Illness Index; Trypsin; Trypsinogen

Local complications of acute pancreatitis (AP) carry risks of morbidity/mortality. This study aimed to assess whether urinary trypsinogen-2 levels and Bedside Index for Severity in Acute Pancreatitis (BISAP) score on admission predicted subsequent local complications.. One hundred and forty-four consecutive patients with AP were prospectively followed till 6 months after discharge. Urinary trypsinogen-2 levels were measured within 24 h of admission. Local complications (acute peripancreatic fluid collection, acute necrotic collection, pseudocyst, and walled-off necrosis) were diagnosed by abdominal computed tomography. Cut-off for trypsinogen-2 level was assessed using receiver operating characteristic curve, and predictors of local complications were analyzed by logistic regression.. Thirty-seven (25.7%) patients developed local complications. Urinary trypsinogen-2 levels were significantly higher in patients with local complications compared with those without local complications (median [interquartile range], 3210 [620-9764.4] μg/L vs 627.3 [72.3-5895] μg/L, P = 0.006). Urinary trypsinogen-2 significantly outperformed BISAP score in predicting local complications (area under the receiver operating characteristic curve 0.65 [95% CI: 0.55-0.75] vs 0.48 [95% CI: 0.38-0.58], P = 0.005). At the optimal cut-off of 500 μg/L, the sensitivity, specificity, positive predictive value, and negative predictive value of trypsinogen-2 level were 78.4%, 45.8%, 33.3%, and 86.0%, respectively. Urinary trypsinogen-2 level > 500 μg/L was an independent predictor of local complications (adjusted odds ratio, 3.72; 95% CI: 1.42-9.76; P = 0.007). By contrast, BISAP score ≥ 3 and pleural effusion predicted organ failure but not local complications.. In a prospective cohort, urinary trypsinogen-2 level > 500 μg/L independently predicted local complications of AP.

Topics: Acute Disease; Biomarkers; Humans; Middle Aged; Pancreatitis; Prospective Studies; Trypsin; Trypsinogen

Premature trypsinogen activation is the early event of acute pancreatitis. Therefore, the studies on the processes of trypsinogen activation induced by compounds are important to understand mechanism underly acute pancreatitis under various conditions. Calcium overload in the early stage of acute pancreatitis was previously found to cause intracellular trypsinogen activation; however, treatment of acute pancreatitis using calcium channel blockers did not produced consistent results. Proteasome activity that could be inhibited by some calcium channel blocker has recently been reported to affect the development of acute pancreatitis; however, the associated mechanism were not fully understood. Here, the roles of nicardipine were investigated in trypsinogen activation in pancreatic acinar cells. The results showed that nicardipine could increase cathepsin B activity that caused trypsinogen activation, but higher concentration of nicardipine or prolonged treatment had an opposite effect. The effects of short time treatment of nicardipine at low concentration were studied here. Proteasome inhibition was observed under nicardipine treatment that contributed to the up-regulation in cytosolic calcium. Increased cytosolic calcium from ER induced by nicardipine resulted in the release and activation of cathepsin B. Meanwhile, calcium chelator inhibited cathepsin B as well as trypsinogen activation. Consistently, proteasome activator protected acinar cells from injury induced by nicardipine. Moreover, proteasome inhibition caused by nicardipine depended on CaMKII. In conclusion, CaMKII down-regulation/proteasome inhibition/cytosolic calcium up-regulation/cathepsin B activation/trypsinogen activation axis was present in pancreatic acinar cells injury under nicardipine treatment.

Topics: Acinar Cells; Animals; Calcium; Calcium Channel Blockers; Calcium Chelating Agents; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Cathepsin B; Cell Line, Tumor; Gene Expression Regulation; Humans; Mice; Nicardipine; Oligopeptides; Pancreas; Pancreatitis; Proteasome Endopeptidase Complex; Trypsinogen

Bile acid causes trypsinogen activation in pancreatic acinar cells through a complex process. Additional research is required to further elucidate which signaling pathways affect trypsinogen activation when activated. the changes in the whole‑genome expression profile of AR42J cells under the effect of taurolithocholic acid 3‑sulfate (TLC‑S) were investigated. Furthermore, gene groups that may play a regulatory role were analyzed using the modular approach of biological networks. The aim of the present study was to improve our understanding of the changes in TLC‑S‑stimulated AR42J cells through a genetic functional modular analysis. whole‑genome expression profile chip arrays were applied to detect genes that were differentially expressed in pancreatic acinar AR42J cells treated with TLC‑S for 20 min. Based on the human protein reference database, a protein‑protein interaction network was obtained, which was then processed by CFinder software to derive 14 modules. Among these 14 modules, the gene ontology biological processes enrichment analysis identified two as modules of interest. Kyoto encyclopedia of genes and genomes map analysis revealed that MAP2K4, MAPK8 and FLNA are part of the c-Jun N-terminal kinase (JNK) pathway. The JNK signaling pathway is involved in regulating trypsinogen activation in rat pancreatic AR42J cells. Next, a regulatory network of seven kinase inhibitors was constructed. SP600125 is an ATP‑competitive, efficient, selective and reversible inhibitor of JNK. the results were verified by four sets of experiments and demonstrated that trypsinogen activation is mediated by the JNK signaling pathway in the pathogenesis of acute pancreatitis (AP). The present study provided a useful reference for better understanding the pathogenesis of AP and identifying new targets to regulate trypsinogen activation, in addition to providing valuable information for the treatment of AP.

Topics: Acinar Cells; Animals; Anthracenes; Epithelial Cells; Gene Expression Regulation; Genome; Humans; JNK Mitogen-Activated Protein Kinases; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 8; Pancreas; Pancreatitis; Protein Interaction Maps; Rats; Signal Transduction; Trypsinogen

There has been recent evidence supporting post-pancreatectomy pancreatitis as a factor in the development of postoperative pancreatic fistula (POPF). The aims of this study were to evaluate: (i) the correlation of the acinar cell density at the pancreatic resection margin with the intra-operative amylase concentration (IOAC) of peri-pancreatic fluid, postoperative pancreatitis, and POPF; and (ii) the association between postoperative pancreatitis on the first postoperative day and POPF.. Consecutive patients who underwent pancreatic resection between June 2016 and July 2017 were included for analysis. Fluid for IOAC was collected, and amylase concentration was determined in drain fluid on postoperative days 1, 3, and 5. Serum amylase and lipase and urinary trypsinogen-2 concentrations were determined on the first postoperative day. Histology slides of the pancreatic resection margin were scored for acinar cell density.. Sixty-one patients were included in the analysis. Acinar cell density significantly correlated with IOAC (r = 0.566, p < 0.001), and was significantly associated with postoperative pancreatitis (p < 0.001), and POPF (p = 0.003). Postoperative pancreatitis was significantly associated with the development of POPF (OR 17.81, 95%CI 2.17-145.9, p = 0.001).. The development of POPF may involve a complex interaction between acinar cell density, immediate leakage of pancreatic fluid, and postoperative pancreatitis.

Topics: Acinar Cells; Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Biopsy; Female; Humans; Lipase; Male; Margins of Excision; Middle Aged; Pancreatectomy; Pancreatic Fistula; Pancreaticoduodenectomy; Pancreatitis; Risk Factors; Time Factors; Treatment Outcome; Trypsin; Trypsinogen

Pancreatic acinar cells are polarized epithelial cells that store enzymes required for digestion as inactive zymogens, tightly packed at the cell apex. Stimulation of acinar cells causes the zymogen granules to fuse with the apical membrane, and the cells undergo exocytosis to release proteases into the intestinal lumen. Autophagy maintains homeostasis of pancreatic acini. Syntaxin 2 (STX2), an abundant soluble N-ethyl maleimide sensitive factor attachment protein receptor in pancreatic acini, has been reported to mediate apical exocytosis. Using human pancreatic tissues and STX2-knockout (KO) mice, we investigated the functions of STX2 in zymogen granule-mediated exocytosis and autophagy.. We obtained pancreatic tissues from 5 patients undergoing surgery for pancreatic cancer and prepared 80-μm slices; tissues were exposed to supramaximal cholecystokinin octapeptide (CCK-8) or ethanol and a low concentration of CCK-8 and analyzed by immunoblot and immunofluorescence analyses. STX2-KO mice and syntaxin 2. Human pancreatic tissues and dispersed pancreatic acini from control mice exposed to CCK-8 or ethanol plus CCK-8 were depleted of STX2. STX2-KO developed more severe pancreatitis after administration of supramaximal caerulein or a 6-week ethanol diet compared with control. Acini from STX2-KO mice had increased apical exocytosis after exposure to CCK-8, as well as increased basolateral exocytosis, which led to ectopic release of proteases. These increases in apical and basolateral exocytosis required increased formation of fusogenic soluble N-ethyl maleimide sensitive factor attachment protein receptor complexes, mediated by STX3 and STX4. STX2 bound ATG16L1 and prevented it from binding clathrin. Deletion of STX2 from acini increased binding of AT16L1 to clathrin, increasing formation of pre-autophagosomes and inducing autophagy. Induction of autophagy promoted the CCK-8-induced increase in autolysosome formation and the activation of trypsinogen.. In studies of human pancreatic tissues and pancreata from STX2-KO and control mice, we found STX2 to block STX3- and STX4-mediated fusion of zymogen granules with the plasma membrane and exocytosis and prevent binding of ATG16L1 to clathrin, which contributes to induction of autophagy. Exposure of pancreatic tissues to CCK-8 or ethanol depletes acinar cells of STX2, increasing basolateral exocytosis and promoting autophagy induction, leading to activation of trypsinogen.

Topics: Acinar Cells; Animals; Autophagy; Cell Membrane; Ceruletide; Exocytosis; Humans; Mice; Mice, Inbred C57BL; Mice, Knockout; Pancreas; Pancreatic Neoplasms; Pancreatitis; Secretory Vesicles; Syntaxin 1; Trypsinogen

Inflammatory diseases of the pancreas have no specific therapy. Discovery of the genetic basis of chronic pancreatitis identified the digestive enzyme trypsin as a therapeutic target. Preclinical testing of trypsin inhibition has been hampered by the lack of animal models. Here we report the T7D23A knock-in mouse, which carries a heterozygous p.D23A mutation in mouse cationic trypsinogen (isoform T7). This trypsinogen mutant autoactivates to trypsin 50-fold faster than wild type. T7D23A mice develop spontaneous acute pancreatitis with edema, necrosis and serum amylase elevation at an early age followed by progressive atrophic chronic pancreatitis with acinar cell loss, fibrosis, dilated ducts and adipose replacement. Markedly elevated trypsin activity is apparent at first signs of pancreatitis and persists into later stages of the disease. This remarkable model provides in vivo proof of concept that trypsinogen autoactivation can drive onset and progression of chronic pancreatitis and therapy should be directed against intra-pancreatic trypsin.

Topics: Animals; Enzyme Activation; Female; Male; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Mutation; Pancreas; Pancreatitis; Pancreatitis, Chronic; Trypsin; Trypsinogen

Topics: Adult; Chymotrypsin; Female; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

The present study was designed to investigate whether the susceptibility to acute pancreatitis (AP) attributable to polymorphism rs10273639 at the PRSS1-PRSS2 locus is dependent on alcohol consumption and cigarette smoking.. A total of 603 unrelated Russian individuals including 304 patients with physician-diagnosed AP and 299 sex- and age-matched healthy controls have been recruited for the study. A polymorphism rs10273639 (-408C>T) of PRSS1-PRSS2 was genotyped by TaqMan-based assay.. A variant allele -408T (P = 0.003) and genotypes -408CT plus TT (P = 0.002) were associated with decreased AP risk only in men. The odds ratios for AP in the CC homozygotes versus the variant genotypes were 1.95 [95% confidence interval (CI), 0.65-5.85; P = 0.23], 1.72 (95% CI, 0.93-3.20; P = 0.08), and 2.37 (95% CI, 1.09-5.13; P = 0.03) for men who consumed up to 28, 29 to 59, and more than 60 alcohol drinks a week, respectively. Cigarette smokers with the -408CC genotype had an increased risk of AP (odds ratio, 2.07; 95% CI, 1.25-3.42; P = 0.004), whereas nonsmoker carriers did not have a disease risk (odds ratio, 1.48; 95% CI, 0.58-3.82; P = 0.42).. We confirmed a robust association of polymorphism rs10273639 at PRSS1-PRSS2 with AP in the Russian population. The present study is the first to show that relationship between the locus and disease is significantly modified by alcohol consumption and cigarette smoking.

Topics: Acute Disease; Adult; Alcohol Drinking; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Humans; Male; Middle Aged; Pancreatitis; Polymorphism, Single Nucleotide; Risk Factors; Sex Factors; Smoking; Trypsin; Trypsinogen

Topics: Chronic Disease; Humans; Mutation; Pancreatitis; Pancreatitis, Chronic; Trypsin; Trypsinogen

Zymogen secretory granules in pancreatic acinar cells express two vesicle-associated membrane proteins (VAMP), VAMP2 and -8, each controlling 50% of stimulated secretion. Analysis of secretion kinetics identified a first phase (0-2 min) mediated by VAMP2 and second (2-10 min) and third phases (10-30 min) mediated by VAMP8. Induction of acinar pancreatitis by supramaximal cholecystokinin (CCK-8) stimulation inhibits VAMP8-mediated mid- and late-phase but not VAMP2-mediated early-phase secretion. Elevation of cAMP during supramaximal CCK-8 mitigates third-phase secretory inhibition and acinar damage caused by the accumulation of prematurely activated trypsin. VAMP8

Topics: Animals; Endosomes; Female; Kinetics; Male; Mice; Mice, Inbred C57BL; Pancreas; Pancreatitis; R-SNARE Proteins; rab5 GTP-Binding Proteins; Rats; Rats, Sprague-Dawley; Trypsin; Trypsinogen; Vesicular Transport Proteins

Topics: Chronic Disease; Humans; Mutation; Pancreatitis; Pancreatitis, Chronic; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The human pancreas expresses two major trypsinogen isoforms, cationic trypsinogen (PRSS1) and anionic trypsinogen (PRSS2). Mutations in PRSS1 cause hereditary pancreatitis by altering cleavage of regulatory nick sites by chymotrypsin C (CTRC) resulting in reduced trypsinogen degradation and increased autoactivation. Despite 90% identity with PRSS1 and a strong propensity for autoactivation, mutations in PRSS2 are not found in hereditary pancreatitis suggesting that activation of this isoform is more tightly regulated. Here, we demonstrated that CTRC promoted degradation and thereby markedly suppressed autoactivation of human anionic trypsinogen more effectively than previously observed with cationic trypsinogen. Increased sensitivity of anionic trypsinogen to CTRC-mediated degradation was due to an additional cleavage site at Leu-148 in the autolysis loop and the lack of the conserved Cys-139-Cys-206 disulfide bond. Significant stabilization of anionic trypsinogen against degradation was achieved by simultaneous mutations of CTRC cleavage sites Leu-81 and Leu-148, autolytic cleavage site Arg-122, and restoration of the missing disulfide bridge. This stands in stark contrast to cationic trypsinogen where single mutations of either Leu-81 or Arg-122 resulted in almost complete resistance to CTRC-mediated degradation. Finally, processing of the trypsinogen activation peptide at Phe-18 by CTRC inhibited autoactivation of anionic trypsinogen, although cationic trypsinogen was strongly stimulated. Taken together, the observations indicate that human anionic trypsinogen is controlled by CTRC in a manner that individual natural mutations are unlikely to increase stability enough to promote intra-pancreatic activation. This unique biochemical property of anionic trypsinogen explains the lack of association of PRSS2 mutations with hereditary pancreatitis.

Topics: Chymotrypsin; Cystine; Enzyme Activation; Enzyme Stability; Humans; Mutation, Missense; Pancreatitis; Protein Processing, Post-Translational; Proteolysis; Trypsin; Trypsinogen

To assess whether pancreatic function is impaired in children with severe acute malnutrition, is different between edematous vs nonedematous malnutrition, and improves by nutritional rehabilitation.. We followed 89 children with severe acute malnutrition admitted to Queen Elizabeth Central Hospital in Blantyre, Malawi. Stool and blood samples were taken on admission and 3 days after initial stabilization to determine exocrine pancreatic function via fecal elastase-1 (FE-1) and serum trypsinogen and amylase levels.. A total of 33 children (37.1%) had nonedematous severe acute malnutrition, whereas 56 (62.9%) had edematous severe acute malnutrition. On admission, 92% of patients showed evidence of pancreatic insufficiency as measured by FE-1 <200 μg/g of stool. Patients with edematous severe acute malnutrition were more likely to have low FE-1 (98% vs 82.8%, P = .026). FE-1 levels remained low in these individuals throughout the assessment period. Serum trypsinogen was elevated (>57 ng/mL) in 28% and amylase in 21% (>110 U/L) of children, suggesting pancreatic inflammation.. Exocrine pancreatic insufficiency is prevalent in children with severe acute malnutrition and especially in children with edematous severe acute malnutrition. In addition, biochemical signs suggestive of pancreatitis are common in children with severe acute malnutrition. These results have implications for standard rehabilitation treatment of children with severe acute malnutrition who may benefit from pancreatic enzyme replacement therapy.. ISRCTN.com: 13916953.

Topics: Amylases; Child, Preschool; Cohort Studies; Exocrine Pancreatic Insufficiency; Female; Humans; Length of Stay; Male; Pancreatic Elastase; Pancreatic Function Tests; Pancreatitis; Prevalence; Severe Acute Malnutrition; Trypsinogen

Previously, we have shown that hydrogen sulphide (H

Topics: Alkynes; AMP-Activated Protein Kinases; Animals; Autophagy; Cell Line; Disease Progression; Glycine; Hydrogen Sulfide; Lysosomes; Male; Pancreatitis; Phagosomes; Rats, Wistar; Signal Transduction; TOR Serine-Threonine Kinases; Trypsinogen; Vacuoles

Topics: Acute Disease; Animals; Autophagy; Extracellular Space; High-Throughput Nucleotide Sequencing; Humans; Neutrophils; Pancreatitis; Trypsinogen

Macrophage activation plays an important role in the inflammatory response in acute pancreatitis. In the present study, the activation of AR42J pancreatic acinar cells was induced by taurolithocholate treatment. The results showed that the culture medium from the activated AR42J cells significantly enhanced NFκB activation in the macrophages compared to that without taurolithocholate treatment. Additionally, the precipitates obtained from ultracentrifugation of the culture media that were rich in exosomes were markedly more potent in activating macrophages compared with the supernatant fraction lacking exosomes. The results indicated that the mediators carried by the exosomes played important roles in macrophage activation. Exosomal miRNAs were extracted and examined using microarrays. A total of 115 differentially expressed miRNAs were identified, and 30 showed upregulated expression, while 85 displayed downregulated expression. Target genes of the differentially expressed miRNAs were predicted using TargetScan, MiRanda, and PicTar software programs. The putative target genes were subjected to KEGG functional analysis. The functions of the target genes were primarily enriched in MAPK pathways. Specifically, the target genes regulated macrophage activation through the TRAF6-TAB2-TAK1-NIK/IKK-NFκB pathway. As the mediators of signal transduction, miRNAs and their predicted target mRNAs regulate every step in the MAPK pathway.

Topics: Acinar Cells; Animals; Cell Line; Cell Survival; Electrophoretic Mobility Shift Assay; Exosome Multienzyme Ribonuclease Complex; Macrophages; MicroRNAs; NF-kappa B; Pancreas; Pancreatitis; Rats; Real-Time Polymerase Chain Reaction; Signal Transduction; Trypsinogen

Pancreatic acinar cell maturation is dependent on the activity of the pancreas transcription factor 1 (PTF1) complex. Induction of pancreatitis leads to MAP kinase activation and transient suppression of the acinar differentiation programme. We investigated the role of MAP kinase-interacting kinase 1 (Mnk1) in mouse exocrine pancreas development and in the response to secretagogue-induced pancreatitis.. Mnk1 expression was analysed using immunohistochemistry, RT-qPCR and western blotting. Ptf1a binding to Mnk1 was assessed by chromatin immunoprecipitation and qPCR. Acute pancreatitis was induced in wild type and Mnk1(-/-) mice by 7 h intraperitoneal injections of caerulein. In vitro amylase secretion and trypsinogen activation were assessed using freshly isolated acinar cells. In vivo secretion was quantified by secretin-stimulated MRI.. Mnk1 is expressed at the highest levels in pancreatic acinar cells and is a direct PTF1 target. Mnk1 is activated upon induction of pancreatitis and is indispensable for eIF4E phosphorylation. The pancreas of Mnk1(-/-) mice is histologically normal. Digestive enzyme content is significantly increased and c-Myc and Ccnd1 levels are reduced in Mnk1(-/-) mice. Upon induction of acute pancreatitis, Mnk1(-/-) mice show impaired eIF4E phosphorylation, activation of c-Myc and downregulation of zymogen content. Acinar cells show defective relocalisation of digestive enzymes, polarity defects and impaired secretory response in vitro and in vivo.. Mnk1 is a novel pancreatic acinar cell-specific stress response kinase that regulates digestive enzyme abundance and eIF4E phosphorylation. It is required for the physiological secretory response of acinar cells and for the homeostatic response to caerulein administration during acute pancreatitis.

Topics: Acinar Cells; Amylases; Animals; Cell Differentiation; Ceruletide; Cholangiopancreatography, Magnetic Resonance; Down-Regulation; Enzyme Activation; Eukaryotic Initiation Factor-4E; Gene Targeting; Heat-Shock Response; Mice; Mitogen-Activated Protein Kinases; Pancreas, Exocrine; Pancreatitis; Phosphorylation; Protein Serine-Threonine Kinases; Transcription Factors; Trypsinogen

We previously reported that atrial natriuretic factor (ANF) stimulates secretin-evoked cAMP efflux through multidrug resistance-associated protein 4 (MRP4) in the exocrine pancreas. Here we sought to establish in vivo whether this mechanism was involved in acute pancreatitis onset in the rat. Rats pretreated with or without probenecid (MRPs general inhibitor) were infused with secretin alone or with ANF. A set of these animals were given repetitive cerulein injections to induce acute pancreatitis. Plasma amylase and intrapancreatic trypsin activities were measured and histological examination of the pancreas performed. Secretin alone activated trypsinogen but induced no pancreatic histological changes. Blockade by probenecid in secretin-treated rats increased trypsin and also induced vacuolization, a hallmark of acute pancreatitis. ANF prevented the secretin response but in the absence of probenecid. In rats with acute pancreatitis, pretreatment with secretin aggravated the disease, but ANF prevented secretin-induced changes. Blockade of MRPs in rats with acute pancreatitis induced trypsinogen activation and larger cytoplasmic vacuoles as well as larger areas of necrosis and edema that were aggravated by secretin but not prevented by ANF. The temporal resolution of intracellular cAMP levels seems critical in the onset of acute pancreatitis, since secretin-evoked cAMP in a context of MRP inhibition makes the pancreas prone to injury in normal rats and aggravates the onset of acute pancreatitis. Present findings support a protective role for ANF mediated by cAMP extrusion through MRP4 and further suggest that the regulation of MRP4 by ANF would be relevant to maintain pancreatic acinar cell homeostasis.

Topics: Acinar Cells; Acute Disease; Animals; Atrial Natriuretic Factor; Cell Membrane; Cyclic AMP; Intracellular Space; Models, Biological; Multidrug Resistance-Associated Proteins; Pancreatitis; Protein Transport; Rats; Trypsinogen

In 126 patients, suffering an acute biliary pancreatitis (ABP), clinical examination was conducted. In 65 patients (1-st group) the isolated cholecystolithiasis was noted; in 35 (2-nd group)--cholelithiasis, which did not cause obturation of common biliary duct; in 26 (3-rd group)--cholelithiasis, which caused the biliary ways obturation (including calculi, which were incorporated into the duodenal papilla magna ostium). Clinical course of an ABP have differed depending on localization of calculi of extrahepatic biliary ducts. In patients, suffering ABP, a biochemical signs of hepatocytes functional disorders were observed, impacting the need for hepatoprotector preparations inclusion into complex of perioperative conservative therapy. Determination of activity of pancreatic α-amylase in the blood serum and conduction of the ACTIM Pancreatitis test con- stitute the most sensitive and specific methods of the ABP biochemical diagnosis.

Topics: Acute Disease; Adult; Alanine Transaminase; Aspartate Aminotransferases; Bile Ducts, Extrahepatic; Cholecystolithiasis; Female; Gallbladder; Glutathione Transferase; Hepatocytes; Humans; Liver; Male; Middle Aged; Pancreas; Pancreatic alpha-Amylases; Pancreatitis; Trypsin; Trypsinogen

Gene polymorphisms, including those recently described in the claudin2 gene, have been implicated in recurrent acute (RAP) and chronic pancreatitis (CP). In India, RAP and CP have been associated with SPINK1 polymorphism. In this study, we evaluated the association of claudin2 and PRSS1-PRSS2 polymorphisms with idiopathic RAP and CP.. We included 101 prospectively followed patients with documented idiopathic RAP (IRAP) and 96 patients who presented with idiopathic chronic pancreatitis (ICP) without previous history of AP. Controls were 156 unrelated individuals undergoing master health check or with non-specific symptoms. All the samples were genotyped for the SNPs rs7057398 in the claudin2 (CLDN2) gene and rs10273639 in the PRSS1 gene on Realtime polymerase chain reaction platform. Clinical data pertaining to patient and disease characteristics were recorded.. Claudin2 and PRSS1 polymorphisms were seen in a significantly higher proportion of female patients (P = 0.01 and 0.039, respectively). Thirty-three (32.7%) patients with IRAP developed features of early CP during follow-up (mean [95% confidence interval, CI] duration of 11.3 [8.9-13.7] months). Female patients with claudin2 (rs7057398) CC genotype were at significantly higher risk for IRAP (odds ratio [OR] [95% CI] 6.75 [1.82-23.67]; P = 0.004) and progression from IRAP to CP (OR [95% CI] 7.05 [1.51-33.01]; P = 0.007). CT genotype of PRSS1 (rs10273639) was associated IRAP (OR [95% CI] 2.59 [1.1-6.13]; P = 0.030), and both CT and CC genotypes with ICP in women (OR [95% CI] 2.86 [1.12-7.31]; P = 0.033 and 3.73 [1.03-13.59]; P = 0.048, respectively).. In this study, we have demonstrated the association of claudin2 (rs7057398) polymorphism with IRAP and progression of IRAP to CP, and PRSS1 (rs10273639) polymorphism with IRAP and ICP.

Topics: Acute Disease; Adolescent; Case-Control Studies; Child; Chronic Disease; Claudin-2; Disease Progression; Female; Follow-Up Studies; Genetic Association Studies; Humans; India; Male; Pancreatitis; Polymorphism, Single Nucleotide; Prospective Studies; Real-Time Polymerase Chain Reaction; Recurrence; Trypsin; Trypsinogen; Young Adult

The signaling mechanisms controlling organ damage in the pancreas in severe acute pancreatitis (AP) remain elusive. Herein, we examined the role of farnesyltransferase signaling in AP.. Pancreatitis was provoked by the infusion of taurocholate into the pancreatic duct in C57BL/6 mice. Animals were treated with a farnesyltransferase inhibitor FTI-277 (25 mg/kg) before pancreatitis induction.. FTI-277 decreased the blood amylase levels, pancreatic neutrophil infiltration, hemorrhage, and edema formation in the pancreas in mice challenged with taurocholate. Farnesyltransferase inhibition reduced the myeloperoxidase levels in the pancreas and lungs in response to taurocholate infusion. However, FTI-277 had no effect on the taurocholate-provoked formation of macrophage inflammatory protein-2 in the pancreas. Interestingly, farnesyltransferase inhibition abolished the neutrophil expression of macrophage-1 antigen in mice with pancreatitis. In addition, FTI-277 decreased the taurocholate-induced activation of the rat sarcoma protein in the pancreas. An important role of farnesyltransferase was confirmed in L-arginine-induced pancreatitis.. These results demonstrate that farnesyltransferase signaling plays a significant role in AP by regulating neutrophil infiltration and tissue injury via the neutrophil expression of macrophage-1 antigen. Thus, our findings not only elucidate novel signaling mechanisms in pancreatitis but also suggest that farnesyltransferase might constitute a target in the management of severe AP.

Topics: Acinar Cells; Acute Disease; Amylases; Animals; Arginine; Cells, Cultured; Chemokine CXCL2; Enzyme Inhibitors; Farnesyltranstransferase; Lung; Macrophage-1 Antigen; Male; Methionine; Mice; Mice, Inbred C57BL; Neutrophil Infiltration; Pancreas; Pancreatitis; Peroxidase; Signal Transduction; Taurocholic Acid; Trypsin; Trypsinogen

Topics: Acute Disease; Animals; Endoplasmic Reticulum; Humans; Immunity, Innate; NF-kappa B; Oxidative Stress; Pancreatitis; Trypsinogen

The strong up-regulation of inflammatory mediators has been reported to play a key role in acute pancreatitis (AP). Elevated serum levels of interleukin-1β (IL-1β) are associated with the development of AP. However, the precise effect and mechanism of IL-1β in AP remains obscure. In this study, we investigated the potential role and mechanism of IL-1β in AP. We measured autophagy activation in response to IL-1β in AR42J cells. The disrupting effects of IL-1β on cellular Ca(2+) were observed. To determine whether the disruption of Ca(2+) signaling has protective effects in vivo during AP, male C57BL/6 mice were treated with cerulein to induce AP. We found that the treatment of AR42J cells with IL-1β triggered autophagy and that the autophagic flux was impaired. In addition, IL-1β induced Ca(2+) release from the ER. Furthermore, the expression of the ER stress markers GRP78 and IRE1 also increased. 2APB, an antagonist of the InsP3 receptor, inhibited increased expression of autophagy markers. Subsequent biochemical assays revealed that co-culture with IL-1β could induce the activation of trypsinogen to trypsin and reduce the viability of acinar cells. Pathological changes of the pancreas were also observed in vivo. We found that the pathological injuries of the pancreas were significantly alleviated in mice co-treated with 2APB. Taken together, our results indicate that IL-1β can induce trypsin activation and decrease cellular viability in pancreatic acinar cells. These effects depend on impaired autophagy via intracellular calcium changes. Ca(2+) signaling may become a promising therapeutic target in the treatment of pancreatitis.

Topics: Acinar Cells; Animals; Autophagy; Blotting, Western; Calcium Signaling; Endoplasmic Reticulum Chaperone BiP; Homeostasis; Interleukin-1beta; Male; Mice; Mice, Inbred C57BL; Microscopy, Confocal; Microscopy, Electron, Transmission; Pancreas; Pancreatitis; Rats; Transfection; Trypsinogen

Leukocyte infiltration and acinar cell necrosis are hallmarks of severe AP, but the signaling pathways regulating inflammation and organ injury in the pancreas remain elusive. In the present study, we investigated the role of geranylgeranyltransferase in AP. Male C57BL/6 mice were treated with a geranylgeranyltransferase inhibitor GGTI-2133 (20 mg/kg) prior to induction of pancreatitis by infusion of taurocholate into the pancreatic duct. Pretreatment with GGTI-2133 reduced plasma amylase levels, pancreatic neutrophil recruitment, hemorrhage, and edema formation in taurocholate-evoked pancreatitis. Moreover, administration of GGTI-2133 decreased the taurocholate-induced increase of MPO activity in the pancreas and lung. Treatment with GGTI-2133 markedly reduced levels of CXCL2 in the pancreas and IL-6 in the plasma in response to taurocholate challenge. Notably, geranylgeranyltransferase inhibition abolished neutrophil expression of Mac-1 in mice with pancreatitis. Finally, inhibition of geranylgeranyltransferase had no direct effect on secretagogue-induced activation of trypsinogen in pancreatic acinar cells in vitro. A significant role of geranylgeranyltransferase was confirmed in an alternate model of AP induced by L-arginine challenge. Our findings show that geranylgeranyltransferase regulates neutrophil accumulation and tissue damage via expression of Mac-1 on neutrophils and CXCL2 formation in AP. Thus, these results reveal new signaling mechanisms in pancreatitis and indicate that targeting geranylgeranyltransferase might be an effective way to ameliorate severe AP.

Topics: Acinar Cells; Acute Disease; Alkyl and Aryl Transferases; Animals; Chemokine CXCL2; Imidazoles; Leucine; Macrophage-1 Antigen; Male; Mice; Mice, Inbred C57BL; Naphthalenes; Neutrophil Infiltration; Neutrophils; Pancreatitis; Prenylation; rac1 GTP-Binding Protein; Trypsinogen

To examine whether rendezvous endoscopic retrograde cholangiopancreatography (ERCP) is associated with less pancreatic damage, measured as leakage of proenzymes, than conventional ERCP.. Patients (n = 122) with symptomatic gallstone disease, intact papilla and no ongoing inflammation, were prospectively enrolled in this case-control designed study. Eighty-one patients were subjected to laparoscopic cholecystectomy and if intraoperative cholangiography suggested common bile duct stones (CBDS), rendezvous ERCP was performed intraoperatively (n = 40). Patients with a negative cholangiogram constituted the control group (n = 41). Another 41 patients with CBDS, not subjected to surgery, underwent conventional ERCP. Pancreatic proenzymes, procarboxypeptidase B and trypsinogen-2 levels in plasma, were analysed at 0, 4, 8 and 24 h. The proenzymes were determined in-house with a double-antibody enzyme linked immunosorbent assay. Pancreatic amylase was measured by an enzymatic colourimetric modular analyser with the manufacturer's reagents. All samples were blinded at analysis.. Post ERCP pancreatitis (PEP) occurred in 3/41 (7%) of the patients cannulated with conventional ERCP and none in the rendezvous group. Increased serum levels indicating pancreatic leakage were significantly higher in the conventional ERCP group compared with the rendezvous ERCP group regarding pancreatic amylase levels in the 4- and 8-h samples (P = 0.0015; P = 0.03), procarboxypeptidase B in the 4- and 8-h samples (P < 0.0001; P < 0.0001) and trypsinogen-2 in the 24-hour samples (P = 0.03). No differences in these markers were observed in patients treated with rendezvous cannulation technique compared with patients that underwent cholecystectomy alone (control group). Post procedural concentrations of pancreatic amylase and procarboxypeptidase B were significantly correlated with pancreatic duct cannulation and opacification.. Rendezvous ERCP reduces pancreatic enzyme leakage compared with conventional ERCP cannulation technique. Thus, laparo-endoscopic technique can be recommended with the ambition to minimise the risk for post ERCP pancreatitis.

Topics: Adult; Aged; Amylases; Biomarkers; Carboxypeptidase B; Case-Control Studies; Catheterization; Chi-Square Distribution; Cholangiopancreatography, Endoscopic Retrograde; Cholecystectomy, Laparoscopic; Colorimetry; Enzyme-Linked Immunosorbent Assay; Female; Gallstones; Humans; Linear Models; Male; Middle Aged; Odds Ratio; Pancreas; Pancreatitis; Prospective Studies; Risk Factors; Time Factors; Treatment Outcome; Trypsin; Trypsinogen

The endocannabinoid system has been shown to mediate beneficial effects on gastrointestinal inflammation via cannabinoid receptors 1 (CB(1)) and 2 (CB(2)). These receptors have also been reported to activate the MAP kinases p38 and c-Jun NH(2)-terminal kinase (JNK), which are involved in early acinar events leading to acute pancreatitis and induction of proinflammatory cytokines. Our aim was to examine the role of cannabinoid receptor activation in an experimental model of acute pancreatitis and the potential involvement of MAP kinases. Cerulein pancreatitis was induced in wild-type, CB(1)-/-, and MK2-/- mice pretreated with selective cannabinoid receptor agonists or antagonists. Severity of pancreatitis was determined by serum amylase and IL-6 levels, intracellular activation of pancreatic trypsinogen, lung myeloperoxidase activity, pancreatic edema, and histological examinations. Pancreatic lysates were investigated by Western blotting using phospho-specific antibodies against p38 and JNK. Quantitative PCR data, Western blotting experiments, and immunohistochemistry clearly show that CB(1) and CB(2) are expressed in mouse pancreatic acini. During acute pancreatitis, an upregulation especially of CB(2) on apoptotic cells occurred. The unselective CB(1)/CB(2) agonist HU210 ameliorated pancreatitis in wild-type and CB(1)-/- mice, indicating that this effect is mediated by CB(2). Furthermore, blockade of CB(2), not CB(1), with selective antagonists engraved pathology. Stimulation with a selective CB(2) agonist attenuated acute pancreatitis and an increased activation of p38 was observed in the acini. With use of MK2-/- mice, it could be demonstrated that this attenuation is dependent on MK2. Hence, using the MK2-/- mouse model we reveal a novel CB(2)-activated and MAP kinase-dependent pathway that modulates cytokine expression and reduces pancreatic injury and affiliated complications.

Topics: Amylases; Animals; Anti-Inflammatory Agents; Apoptosis; Blotting, Western; Cannabinoids; Ceruletide; Disease Models, Animal; Dronabinol; Edema; Enzyme Activation; Immunohistochemistry; Interleukin-6; Intracellular Signaling Peptides and Proteins; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; p38 Mitogen-Activated Protein Kinases; Pancreas, Exocrine; Pancreatitis; Peroxidase; Phosphorylation; Polymerase Chain Reaction; Protein Serine-Threonine Kinases; Receptor, Cannabinoid, CB1; Receptor, Cannabinoid, CB2; Trypsinogen

Pancreatic acinar cells express proteinase-activated receptor-2 (PAR2) that is activated by trypsin-like serine proteases and has been shown to exert model-specific effects on the severity of experimental pancreatitis, i.e., PAR2(-/-) mice are protected from experimental acute biliary pancreatitis but develop more severe secretagogue-induced pancreatitis. P2pal-18S is a novel pepducin lipopeptide that targets and inhibits PAR2. In studies monitoring PAR2-stimulated intracellular Ca(2+) concentration changes, we show that P2pal-18S is a full PAR2 inhibitor in acinar cells. Our in vivo studies show that P2pal-18S significantly reduces the severity of experimental biliary pancreatitis induced by retrograde intraductal bile acid infusion, which mimics injury induced by endoscopic retrograde cholangiopancreatography (ERCP). This reduction in pancreatitis severity is observed when the pepducin is given before or 2 h after bile acid infusion but not when it is given 5 h after bile acid infusion. Conversely, P2pal-18S increases the severity of secretagogue-induced pancreatitis. In vitro studies indicate that P2pal-18S protects acinar cells against bile acid-induced injury/death, but it does not alter bile acid-induced intracellular zymogen activation. These studies are the first to report the effects of an effective PAR2 pharmacological inhibitor on pancreatic acinar cells and on the severity of experimental pancreatitis. They raise the possibility that a pepducin such as P2pal-18S might prove useful in the clinical management of patients at risk for developing severe biliary pancreatitis such as occurs following ERCP.

Topics: Acinar Cells; Animals; Bile Acids and Salts; Biliary Tract Diseases; Calcium; Calcium Signaling; Ceruletide; Cholangiopancreatography, Endoscopic Retrograde; Chymotrypsinogen; Coloring Agents; Enzyme Activation; Enzyme Precursors; Gallstones; Indicators and Reagents; Lipopeptides; Mice; Mice, Inbred C57BL; Mice, Knockout; Pancreatitis; Propidium; Receptor, PAR-2; Trypsinogen

MMPs are generally considered to regulate degradation and remodeling of the ECM. Convincing data also implicate a role for MMPs in inflammatory conditions, such as AP, although the mechanisms are not known. The aim of this study was to define the role of MMPs in regulating activation of trypsinogen and tissue damage in AP, which was induced by infusion of taurocholate into the pancreatic duct in mice. A broad-spectrum MMP inhibitor (BB-94) and MMP-9 gene-deficient mice were used. Neutrophil secretions and rMMP-9 were used to stimulate trypsinogen activation in isolated acinar cells. Taurocholate challenge increased serum amylase, neutrophil infiltration, MIP-2 (CXCL2) formation, trypsinogen activation, and tissue damage in the pancreas. Treatment with the broad-spectrum inhibitor of MMPs, BB-94, markedly reduced activation of trypsinogen, levels of CXCL2, infiltration of neutrophils, and tissue damage in AP. Taurocholate challenge increased serum levels of MMP-9 but not MMP-2. Taurocholate-induced amylase levels, neutrophil accumulation, production of CXCL2, trypsinogen activation, and tissue damage in the pancreas were abolished in MMP-9-deficient mice. Moreover, secretions from activated neutrophils isolated from WT but not from MMP-9-deficient animals stimulated trypsinogen activation in acinar cells. Notably, rMMP-9 greatly enhanced activation of trypsinogen in acinar cells. These findings demonstrate that neutrophil-derived MMP-9 is a potent activator of trypsinogen in acinar cells and regulates pathological inflammation and tissue damage in AP.

Topics: Acinar Cells; Acute Disease; Animals; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; Pancreatitis; Peroxidase; Taurocholic Acid; Trypsinogen

The adhesive mechanisms regulating leucocyte-endothelium interactions in the pancreas remain elusive, but selectins may play a role. This study examined the molecular mechanisms mediating leucocyte rolling along the endothelium in the pancreas and the therapeutic potential of targeting the rolling adhesive interaction in acute pancreatitis (AP).. Pancreatitis was induced by retrograde infusion of 5 per cent sodium taurocholate into the pancreatic duct, repeated intraperitoneal administration of caerulein (50 µg/kg) or intraperitoneal administration of L-arginine (4 g/kg) in C57BL/6 mice. A control and a monoclonal antibody against P-selectin were administered before and after induction of AP. Serum and tissue were sampled to assess the severity of pancreatitis, and intravital microscopy was used to study leucocyte rolling.. Taurocholate infusion into the pancreatic duct increased the serum level of trypsinogen, trypsinogen activation, pancreatic neutrophil infiltration, macrophage inflammatory protein (MIP) 2 formation and tissue damage. Immunoneutralization of P-selectin decreased the taurocholate-induced increase in serum trypsinogen (median (range) 17·35 (12·20-30·00) versus 1·55 (0·60-15·70) µg/l; P = 0·017), neutrophil accumulation (4·00 (0·75-4·00) versus 0·63 (0-3·25); P = 0·002) and tissue damage, but had no effect on MIP-2 production (14·08 (1·68-33·38) versus 3·70 (0·55-51·80) pg/mg; P = 0·195) or serum trypsinogen activating peptide level (1·10 (0·60-1·60) versus 0·45 (0-1·80) µg/l; P = 0·069). Intravital fluorescence microscopy revealed that anti-P-selectin antibody inhibited leucocyte rolling completely in postcapillary venules of the inflamed pancreas.. Inhibition of P-selectin protected against pancreatic tissue injury in experimental pancreatitis. Targeting P-selectin may be an effective strategy to ameliorate inflammation in AP.

Topics: Acute Disease; Animals; Cell Adhesion; Chemokine CXCL2; Cholagogues and Choleretics; Cytokines; Endothelium; Leukocyte Count; Leukocyte Rolling; Male; Mice; Mice, Inbred C57BL; Neutrophils; P-Selectin; Pancreatitis; Peroxidase; Taurocholic Acid; Trypsinogen

A simple urinary trypsinogen-2 test was evaluated for the diagnosis of acute pancreatitis.. This prospective multicenter study enrolled consecutive patients with acute abdominal pain who presented to the emergency department or who were hospitalized at 1 of 21 medical institutions in Japan. Patients were tested with urinary trypsinogen-2 dipstick test and a quantitative trypsinogen-2 assay, and these values were compared with serum amylase and lipase findings.. A total of 412 patients were enrolled. The trypsinogen-2 dipstick test was positive in 107 of 156 patients with acute pancreatitis (sensitivity, 68.6%) and in 33 of 256 patients with nonpancreatic abdominal pain (specificity, 87.1%). The sensitivity for the diagnosis of pancreatitis caused by alcohol and gallstones by the dipstick test was 72.2% and 81.8%, respectively, which was much higher than those associated with amylase testing. There are several degrees of positivity within the urinary trypsinogen-2 dipstick test. Modification of the cutoff point such that positive (+) and most positive (++) results were interpreted as a positive result, the specificity and positive likelihood ratio increased to 92.2% and 7.63, respectively.. This simple, rapid, easy, and noninvasive urinary trypsinogen-2 test can diagnose or rule out most cases of acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Chi-Square Distribution; Clinical Enzyme Tests; Female; Humans; Japan; Lipase; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Prospective Studies; Reagent Strips; Reproducibility of Results; Sensitivity and Specificity; Trypsin; Trypsinogen; Urinalysis; Young Adult

Topics: Acinar Cells; Animals; Matrix Metalloproteinase 9; Neutrophils; Pancreatitis; Trypsinogen

Several studies have suggested that autophagy might play a deleterious role in acute pancreatitis via intra-acinar activation of digestive enzymes. The prototype for this phenomenon is cathepsin B-mediated trypsin generation. To determine the organellar basis of this process, we investigated the subcellular distribution of the cathepsin B precursor, procathepsin B. We found that procathepsin B is enriched in Golgi-containing microsomes, suggesting a role for the ADP-ribosylation (ARF)-dependent trafficking of cathepsin B. Indeed, caerulein treatment increased processing of procathepsin B, whereas a known ARF inhibitor brefeldin A (BFA) prevented this. Similar treatment did not affect processing of procathepsin L. BFA-mediated ARF1 inhibition resulted in reduced cathepsin B activity and consequently reduced trypsinogen activation. However, formation of light chain 3 (LC3-II) was not affected, suggesting that BFA did not prevent autophagy induction. Instead, sucrose density gradient centrifugation and electron microscopy showed that BFA arrested caerulein-induced autophagosomal maturation. Therefore, ARF1-dependent trafficking of procathepsin B and the maturation of autophagosomes results in cathepsin B-mediated trypsinogen activation induced by caerulein.

Topics: ADP-Ribosylation Factor 1; Animals; Autophagy; Blotting, Western; Brefeldin A; Cathepsin B; Ceruletide; Enzyme Precursors; Golgi Apparatus; Mice; Microscopy, Electron; Microscopy, Fluorescence; Pancreatitis; Rats; Real-Time Polymerase Chain Reaction; Trypsinogen

Soft pancreas is considered as a factor for pancreatitis after pancreaticoduodenectomy, which in turn constitutes a high risk for local complications. The aim was to analyze the proportion of different cell types in the cut edge of pancreas (CEP) in relation to postoperative pancreatitis and other complications after pancreaticoduodenectomy.. Data from postoperative follow-up was collected on 40 patients who had undergone pancreaticoduodenectomy. Positive urine trypsinogen-2, an early detector of pancreatitis, was checked on days 1 to 6 after operation. Drain amylase was measured on postoperative day 3. Anastomotic leakages, delayed gastric emptying, and other complications were registered. The areas of different cell types were calculated from the entire hematoxylin-eosin-stained section of CEP.. High frequency of acinar cells in the CEP significantly increased positive urine trypsinogen-2 days, drain amylase values, and delayed gastric emptying. In a subgroup of patients with more than 40% acini in the CEP, there were significantly more postoperative complications. Increased fibrosis correlated with a small number of positive urine trypsinogen-2 days and postoperative complications.. A large number of acinar cells in the CEP increases, whereas extensive fibrosis in the CEP decreases, the risk for postoperative complications after pancreaticoduodenectomy. These results emphasize the importance of acini in the development of postoperative complications.

Topics: Acinar Cells; Adult; Aged; Aged, 80 and over; Amylases; Anastomotic Leak; Biomarkers; Chi-Square Distribution; Female; Fibrosis; Finland; Gastroparesis; Humans; Male; Middle Aged; Pancreas; Pancreaticoduodenectomy; Pancreatitis; Retrospective Studies; Risk Assessment; Risk Factors; Time Factors; Treatment Outcome; Trypsin; Trypsinogen; Young Adult

The signaling mechanisms that regulate trypsinogen activation and inflammation in acute pancreatitis (AP) are unclear. We explored the involvement of the calcium- and calcineurin-dependent transcription factor nuclear factor of activated T cells (NFAT) in development of AP in mice.. We measured levels of myeloperoxidase and macrophage inflammatory protein 2 (CXCL2), trypsinogen activation, and tissue damage in the pancreas 24 hours after induction of AP by retrograde infusion of taurocholate into the pancreatic ducts of wild-type, NFAT luciferase reporter (NFAT-luc), and NFATc3-deficient mice. We isolated acinar cells and measured NFAT nuclear accumulation, trypsin activity, and expression of NFAT-regulated genes.. Infusion of taurocholate increased the transcriptional activity of NFAT in the pancreas, aorta, lung, and spleen of NFAT-luc mice. Inhibition of NFAT with A-285222 blocked taurocholate-induced activation of NFAT in all organs. A-285222 also reduced taurocholate-induced increases in levels of amylase, myeloperoxidase, and CXCL2; activation of trypsinogen; necrosis of acinar cells; edema; leukocyte infiltration; and hemorrhage in the pancreas. NFATc3-deficient mice were protected from these effects of taurocholate. Similar results were obtained using an l-arginine-induced model of AP. Reverse-transcription polymerase chain reaction and confocal immunofluorescence analyses showed that NFATc3 is expressed by acinar cells. NFATc3 expression was activated by stimuli that increase intracellular calcium levels, and activation was prevented by the calcineurin blocker cyclosporin A or A-285222. Activation of trypsinogen by secretagogues in acinar cells was prevented by pharmacologic inhibition of NFAT signaling or lack of NFATc3. A-285222 also reduced expression of inflammatory cytokines such as CXCL2 in acinar cells.. NFATc3 regulates trypsinogen activation, inflammation, and pancreatic tissue damage during development of AP in mice and might be a therapeutic target.

Topics: Acinar Cells; Amylases; Animals; Aorta; Cell Nucleus; Chemokine CXCL2; Lung; Mice; Neutrophils; NFATC Transcription Factors; Pancreatitis; Peroxidase; Pyrazoles; Signal Transduction; Spleen; Statistics, Nonparametric; Taurocholic Acid; Trypsinogen

Topics: Acute Disease; Adolescent; Adult; Calibration; Female; Fluoroimmunoassay; Humans; Male; Middle Aged; Pancreatitis; Postprandial Period; Reference Values; Trypsin; Trypsinogen; Young Adult

The role of endothelins in acute pancreatitis remains obscure. To assess the effects of endothelins (ETs) in early (4 h) caerulein-induced acute pancreatitis (AP) in rats, ET-1, ET-2 and ET-3 (0.5 or 1.0 nmol/kg) were applied twice with i.p. caerulein (2×40 μg/kg) at 1h interval. Histological and ultrastructural examinations of pancreases and the assay of trypsinogen activation in whole homogenate were performed. All ETs, especially ET-1 at the higher dose, decreased inflammatory cell infiltration despite an increase in the edema score. The vacuolization and necrosis of acinar cells were slightly increased after the lower dose of ET-1 and ET-2. Ultrastructural changes were generally improved after the higher dose of ETs. Trypsinogen activation increased from 4.8±1.3% in control to 18.4±3.8% in AP (p<0.01). It was attenuated to 6.4±1.3% (p<0.01) by the higher dose of ET-1 and to 8.8±1.5% (p<0.05) by the lower dose of ET-3. In summary, ETs, especially ET-1 at the higher dose, were found to have some beneficial effects on morphological changes and trypsinogen activation in the pancreas in early caerulein-induced AP.

Topics: Animals; Ceruletide; Dose-Response Relationship, Drug; Endothelins; Enzyme Activation; Male; Microscopy, Electron, Transmission; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

A relationship between primary hyperparathyroidism (pHPT) and pancreatitis has long been debated and remains a rare epiphenomenon. In a cohort of patients with pHPT and pancreatitis mutations in the serine protease inhibitor Kazal type I (SPINK1) and cystic fibrosis transmembrane conductance regulator (CFTR) genes, that increase the risk for pancreatitis have already been detected. Among the identification of additional pancreatitis-associtated mutations in the Chymotrypsin C gene (CTRC) it became clear that also protective genetic variants exist in the anionic trypsinogen gene (PRSS2) that decrease susceptibility for pancreatitis. Our aim was to detect either protective or inducing genetic factors in a large cohort of pHPT patients.. Among 1,259 patients with pHPT, 57 patients were identified with pancreatitis (4.5%). DNA was available from 31 patients (16 acute pancreatitis/15 chronic pancreatitis). These individuals and 100 patients with pHPT without pancreatitis were analysed for CTRC (p.R254W and p.K247_R254del) and PRSS2 (p.G191R) mutations using melting curve analysis and DNA sequencing or PCR and gel electrophoresis (in case of p.K247_R254del CTRC).. 2 of 31 patients with pHPT and pancreatitis carried the CTRC p.R254W missense mutation (6.5%), while all 100 pHPT controls without pancreatitis showed no CTRC mutation (P=0.055). No further SPINK1 p.N34S (n=4) mutations were detected but the probability of either CTRC or SPINK1 mutations in pHPT patients with pancreatitis is high (P<0.05). 1 patient was trans-heterozygous ( SPINK1: N34S/ CTRC p.R254W). CTRC p.K247_R254del was not detected in both groups. PRSS2 (p.G191R) mutation was present in 1 patient with pancreatitis (3.2%) and in 6 pHPT controls (6%) (P=1).. This study underlines the relevance of a genetic background in pHPT related pancreatitis. However, it only indicates that the CTRC (p.R254W) mutation might also contribute to the panel of mutations ( SPINK1 and CFTR) that have been formerly reported to elevate pancreatitis susceptibility in pHPT. Besides it suggests that protective genetic variants, i. e., p.G191R PRSS2, may contribute to the low prevalence of pancreatitis in pHPT patients.

Topics: Aged; Chymotrypsin; Databases, Factual; DNA Mutational Analysis; Female; Genetic Association Studies; Genetic Predisposition to Disease; Humans; Hyperparathyroidism, Primary; Male; Middle Aged; Pancreatitis; Polymerase Chain Reaction; Trypsin; Trypsinogen

Severe acute pancreatitis (SAP) is characterized by trypsinogen activation, infiltration of leucocytes and tissue necrosis but the intracellular signalling mechanisms regulating organ injury in the pancreas remain elusive. Rho-kinase is a potent regulator of specific cellular processes effecting several pro-inflammatory activities. Herein, we examined the role of Rho-kinase signalling in acute pancreatitis.. Pancreatitis was induced by infusion of taurocholate into the pancreatic duct in C57BL/6 mice. Animals were treated with a Rho-kinase inhibitor Y-27632 (0.5-5 mg·kg⁻¹) before induction of pancreatitis.. Taurocholate infusion caused a clear-cut increase in blood amylase, pancreatic neutrophil infiltration, acinar cell necrosis and oedema formation in the pancreas. Levels of pancreatic myeloperoxidase (MPO), macrophage inflammatory protein-2 (MIP-2), trypsinogen activation peptide (TAP) and lung MPO were significantly increased, indicating local and systemic disease. Inhibition of Rho-kinase activity dose-dependently protected against pancreatitis. For example, 5 mg·kg⁻¹ Y-27632 reduced acinar cell necrosis, leucocyte infiltration and pancreatic oedema by 90%, 89% and 58%, respectively, as well as tissue levels of MPO by 75% and MIP-2 by 84%. Moreover, Rho-kinase inhibition decreased lung MPO by 75% and blood amylase by 83%. Pancreatitis-induced TAP levels were reduced by 61% in Y-27632-treated mice. Inhibition of Rho-kinase abolished secretagogue-induced activation of trypsinogen in pancreatic acinar cells in vitro.. Our novel data suggest that Rho-kinase signalling plays an important role in acute pancreatitis by regulating trypsinogen activation and subsequent CXC chemokine formation, neutrophil infiltration and tissue injury. Thus, these results indicate that Rho-kinase may constitute a novel target in the management of SAP.

Topics: Amides; Amylases; Animals; Chemokine CXCL2; Cholagogues and Choleretics; Enzyme Inhibitors; Flow Cytometry; Male; Mice; Mice, Inbred C57BL; Necrosis; Neutrophil Infiltration; Pancreas; Pancreatitis; Peroxidase; Pyridines; rho-Associated Kinases; Signal Transduction; Taurocholic Acid; Trypsinogen

Previous studies have shown an association of variants in trypsin-associated genes, such as cationic trypsinogen (PRSS1) and serine protease inhibitor, Kazal type-1 (SPINK1) with pancreatitis. However, whether these genetic variants are associated with acute pancreatitis (AP) remains largely unknown, especially when the first attack is separated from recurrent attacks.. A total of 261 patients with AP (174 with a sentinel attack, and 87 with recurrent attacks) and healthy controls were genotyped for the p.R122H mutation in the PRSS1 gene, p.N34S and IVS3 + 2T > C variants in the SPINK1 gene, the p.G191R variant in the anionic trypsinogen gene, the p.E32del variant in the mesotrypsinogen (PRSS3) gene, and the -2518G > A variant in the monocyte chemoattractant protein-1 gene by polymerase chain reaction-restriction enzyme digestion and direct sequencing.. Patients with recurrent attacks were younger. The proportions of biliary pancreatitis and severe cases were lower, and that of idiopathic pancreatitis was higher in patients with a sentinel attack than in those with recurrent attacks. The frequencies of the genetic variants examined did not differ between controls and patients with sentinel pancreatitis. The frequencies of the PRSS1 p.R122H mutation, SPINK1 p.N34S variant, and PRSS3 p.E32del variant, but not other genetic variants, were higher in patients with recurrent attacks than in controls or those with a sentinel attack.. The PRSS1 p.R122H mutation, SPINK1 p.N34S, and PRSS3 p.E32del variants were associated with recurrent, but not sentinel AP. The genetic background could possibly be different between sentinel and recurrent AP.

Topics: Acute Disease; Adult; Aged; Carrier Proteins; Case-Control Studies; Chemokine CCL2; Chi-Square Distribution; DNA Mutational Analysis; Female; Gene Frequency; Genetic Predisposition to Disease; Humans; Japan; Male; Middle Aged; Mutation; Odds Ratio; Pancreatitis; Phenotype; Recurrence; Risk Assessment; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen; Young Adult

Overinduced polyamine catabolism (PC) in a transgenic rat model has been suggested to be a mediator of trypsin activation which is important in acinar cell necrosis. PC has also been observed in experimental taurodeoxycholate pancreatitis. We hypothesized that PC may be a mediator of trypsin activation in taurodeoxycholate pancreatitis.. Pancreatitis was induced in wild-type rats by 2 or 6% taurodeoxycholate infusion or in transgenic rats by overexpressing spermidine/spermine N(1)-acetyltransferase (SSAT). The time courses of necrosis, caspase-3 immunostaining, SSAT, polyamine levels, and trypsinogen activation peptide (TAP) were monitored. The effect of the polyamine analogue bismethylspermine (Me(2)Spm) was investigated.. In a transgenic pancreatitis model, TAP and acinar necrosis increased simultaneously after the activation of SSAT, depletion of spermidine, and development of apoptosis. In taurodeoxycholate pancreatitis, necrosis developed along with the accumulation of TAP. SSAT was activated simultaneously or after TAP accumulation and less than in the transgenic model, with less depletion of spermidine than in the transgenic model. Supplementation with Me(2)Spm ameliorated the extent of acinar necrosis at 24 h, but contrary to previous findings in the transgenic model, in the taurodeoxycholate model it did not affect trypsin activation. Compared with the transgenic model, no extensive apoptosis was found in taurodeoxycholate pancreatitis.. Contrary to transgenic SSAT-overinduced pancreatitis, PC may not be a mediator of trypsin activation in taurodeoxycholate pancreatitis. The beneficial effect of polyamine supplementation on necrosis in taurodeoxycholate pancreatitis may rather be mediated by other mechanisms than amelioration of trypsin activation. and IAP.

Topics: Acetyltransferases; Animals; Apoptosis; Disease Models, Animal; Enzyme Activation; Male; Oligopeptides; Pancreatitis; Polyamines; Rats; Rats, Sprague-Dawley; Rats, Transgenic; Rats, Wistar; Spermine; Taurodeoxycholic Acid; Trypsin; Trypsinogen

To assess a point-of-care urine trypsinogen-2 (UT) test for the diagnosis of acute pancreatitis.. This was a prospective study of patients presenting to the emergency department with abdominal pain suggestive of acute pancreatitis. A 3-minute point-of-care UT test (Actim Pancreatitis; Medix Biochemica, Kauniainen, Finland) was compared with final diagnosis of acute pancreatitis, which was based on suggestive clinical features, serum lipase and/or amylase levels and imaging.. Of 124 patients included in this study, 69 patients had final diagnosis of acute pancreatitis. The sensitivity and specificity of UT were, respectively, 73.9% (95% CI 61.9% to 83.8%) and 94.6% (95% CI 84.9% to 98.9%).. The point-of-care UT test for acute pancreatitis had good sensitivity and specificity, and can be used reliably at the bedside to make a positive diagnosis.

Topics: Abdomen, Acute; Acute Disease; Amylases; Humans; India; Lipase; Pancreatitis; Patients; Point-of-Care Systems; Prospective Studies; Reproducibility of Results; Sensitivity and Specificity; Trypsin; Trypsinogen

The objective of the present study was to assess the use of serum trypsinogen-2 (TRY-2) measurements in early diagnosis of pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).. In this prospective study, investigation 1 involved collection of blood serum both before and at 2, 4, and 18 hours after ERCP, whereas investigation 2 involved collection before and 1, 2, 3, 4, 6, and 18 hours after ERCP. Total amylase, pancreatic amylase, and TRY-2 levels were measured from serum samples, and values from patients with pancreatitis after ERCP were compared to those from healthy control patients after ERCP.. In investigation 1, 8 of the 68 cases examined were diagnosed as post-ERCP pancreatitis. In the healthy group, total- and pancreatic-amylase levels peaked 4 hours after ERCP, and TRY-2 levels peaked at 2 hours after ERCP. In contrast, cases of post-ERCP pancreatitis demonstrated prolonged periods of high total-amylase, pancreatic-amylase, and TRY-2 levels. In investigation 2, none of the 23 cases was diagnosed as post-ERCP pancreatitis: Pancreatic amylase levels peaked 4 to 6 hours after ERCP and TRY-2 levels peaked 1 hour after ERCP.. These results suggest that TRY-2 is a more sensitive marker than amylase, and it can be useful in early diagnosis of post-ERCP pancreatitis.

Topics: Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Cholangiopancreatography, Endoscopic Retrograde; Early Diagnosis; Female; Humans; Male; Middle Aged; Pancreatitis; Prospective Studies; Sensitivity and Specificity; Time Factors; Trypsin; Trypsinogen

The relationship between inflammation and proteolytic activation in pancreatitis is an unresolved issue in pancreatology. The purpose of this study was to define the influence of neutrophils on trypsinogen activation in severe AP. Pancreatitis was induced by infusion of taurocholate into the pancreatic duct in C57BL/6 mice. For neutrophil depletion, an anti-Gr-1 antibody was administered before pancreatitis induction. Administration of the anti-Gr-1 antibody reduced circulating neutrophils by 97%. Pancreatic TAP and serum amylase levels increased 2 h and 24 h after induction of pancreatitis. Neutrophil depletion reduced pancreatic TAP and serum amylase levels at 24 h but not at 2 h after pancreatitis induction. Pancreatic MPO and infiltration of neutrophils, as well as MIP-2 levels, were increased 24 h after taurocholate infusion. Two hours after taurocholate administration, no significant pancreatic infiltration of neutrophils was observed. Injection of the anti-Gr-1 antibody abolished MPO activity, neutrophil accumulation, and MIP-2 levels, as well as acinar cell necrosis, hemorrhage, and edema in the pancreas at 24 h. Moreover, taurocholate-provoked tissue damage and MPO activity in the lung were normalized by neutrophil depletion. Intravital fluorescence microscopy revealed a 97% reduction of leukocytes in the pancreatic microcirculation after administration of the anti-Gr-1 antibody. Our data demonstrate that initial trypsinogen activation is independent of neutrophils, whereas later activation is dependent on neutrophils in the pancreas. Neutrophils are critical in mediating pancreatic and lung tissue damage in severe AP.

Topics: Acinar Cells; Acute Disease; Amylases; Animals; Cholagogues and Choleretics; Enzyme Activation; Mice; Mice, Inbred C57BL; Neutrophil Activation; Neutrophils; Pancreatitis; Taurocholic Acid; Trypsinogen

The role of trypsinogen activation in the pathogenesis of acute pancreatitis (AP) has not been clearly established.. We generated and characterized mice lacking trypsinogen isoform 7 (T7) gene (T(-/-)). The effects of pathologic activation of trypsinogen were studied in these mice during induction of AP with cerulein. Acinar cell death, tissue damage, early intra-acinar activation of the transcription factor nuclear factor κB (NF-κB), and local and systemic inflammation were compared between T(-/-) and wild-type mice with AP.. Deletion of T7 reduced the total trypsinogen content by 60% but did not affect physiologic function. T(-/-) mice lacked pathologic activation of trypsinogen, which occurs within acinar cells during early stages of AP progression. Absence of trypsinogen activation in T(-/-) mice led to near complete inhibition of acinar cell death in vitro and a 50% reduction in acinar necrosis during AP progression. However, T(-/-) mice had similar degrees of local and systemic inflammation during AP progression and comparable levels of intra-acinar NF-κB activation, which was previously shown to occur concurrently with trypsinogen activation during early stages of pancreatitis.. T7 is activated during pathogenesis of AP in mice. Intra-acinar trypsinogen activation leads to acinar death during early stages of pancreatitis, which accounts for 50% of the pancreatic damage in AP. However, progression of local and systemic inflammation in AP does not require trypsinogen activation. NF-κB is activated early in acinar cells, independently of trypsinogen activation, and might be responsible for progression of AP.

Topics: Acinar Cells; Animals; Cell Death; Enzyme Activation; Inflammation; Isoenzymes; Mice; Mice, Inbred C57BL; Mice, Knockout; NF-kappa B; Pancreatitis; Trypsinogen

The aim of this prospective study was to evaluate the diagnostic value of the rapid urinary trypsinogen-2 test strip in post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis.. A total of 150 patients were tested with the urinary trypsinogen-2 test strip and serum levels of amylase and lipase before ERCP and 3 hours after ERCP. The diagnostic value of urinary trypsinogen-2 strip test compared with that of serum amylase and lipase was analyzed.. Post-ERCP pancreatitis was diagnosed in 13 (8.7%) of 150 patients. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of urinary trypsinogen-2 dipstick test at 3 hours after ERCP are 84.6%, 97.1%, 73.3%, 98.5%, and 96%, respectively. At the cutoff level of 3 times the upper reference limit, the negative predictive values of amylase and lipase were comparable to that urinary trypsinogen-2 strip test; however, their positive predictive values (42.9% and 36.4%, respectively) were markedly lower than that of urinary trypsinogen-2 test (73.3%).. The urinary trypsinogen-2 dipstick test is a useful test for early diagnosis of post-ERCP pancreatitis. A negative urinary dipstick test at 3 hours after the procedure rules out post-ERCP pancreatitis with a high probability and allows of early discharge plan.

Topics: Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Cholangiopancreatography, Endoscopic Retrograde; Early Diagnosis; Female; Humans; Lipase; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Time Factors; Trypsin; Trypsinogen

Topics: Acinar Cells; Animals; Pancreatitis; Trypsinogen

Mice deficient for interferon regulatory factor (Irf)2 (Irf2(-/-) mice) exhibit immunological abnormalities and cannot survive lymphocytic choriomeningitis virus infection. The pancreas of these animals is highly inflamed, a phenotype replicated by treatment with poly(I:C), a synthetic double-stranded RNA. Trypsinogen5 mRNA was constitutively up-regulated about 1,000-fold in Irf2(-/-) mice compared with controls as assessed by quantitative RT-PCR. Further knockout of IFNα/β receptor 1(Ifnar1) abolished poly(I:C)-induced pancreatitis but had no effect on the constitutive up-regulation of trypsinogen5 gene, indicating crucial type I IFN signaling to elicit the inflammation. Analysis of Ifnar1(-/-) mice confirmed type I IFN-dependent transcriptional activation of dsRNA-sensing pattern recognition receptor genes MDA5, RIG-I, and TLR3, which induced poly(I:C)-dependent cell death in acinar cells in the absence of IRF2. We speculate that Trypsin5, the trypsinogen5 gene product, leaking from dead acinar cells triggers a chain reaction leading to lethal pancreatitis in Irf2(-/-) mice because it is resistant to a major endogenous trypsin inhibitor, Spink3.

Topics: Acinar Cells; Animals; Cathepsin B; Glycoproteins; HEK293 Cells; HeLa Cells; Humans; Interferon Regulatory Factor-2; Mice; Mice, Transgenic; Pancreatitis; Poly I-C; Prostatic Secretory Proteins; RNA, Double-Stranded; Transcription, Genetic; Trypsin Inhibitor, Kazal Pancreatic; Trypsin Inhibitors; Trypsinogen

In acute pancreatitis (AP), rapid diagnosis and early treatment are of importance for clinical outcome. Urinary trypsinogen-2 has been suggested as a promising diagnostic marker; however, studies using the urinary trypsinogen-2 dipstick test (UTDT) have provided varying results.. The study was set to evaluate the use of the UTDT (Actim Pancreatitis; Medix Biochemica, Kauniainen, Finland, Medinor, Roskilde, Denmark) in apparent first attack of AP in daily clinics. Acute pancreatitis was defined as more than a 3-fold increase in plasma amylase levels. We included 75 patients admitted with AP. Thirty-four patients with acute abdominal pain of causes other than AP served as a control group.. In 58 of 75 patients, the UTDT result was positive, giving a sensitivity of 77% (95% confidence interval [CI]: 66%-86%). In severe cases, the sensitivity improved to 87% (95% CI: 69%-96%). In 33 of 34 controls, the test result was negative, giving a specificity of 97% (95% CI: 84%-99.9%).. The UTDT had a low sensitivity but high specificity. These results do not support the UTDT to replace standard plasma amylase for the diagnosis of apparent first attack of AP. However, the test demonstrated an adequate sensitivity to be used for rapid early screening of AP in daily clinics.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; Diagnosis, Differential; Female; Humans; Male; Middle Aged; Pancreatitis; Prospective Studies; Reagent Kits, Diagnostic; Reagent Strips; Reproducibility of Results; Sensitivity and Specificity; Trypsin; Trypsinogen; Young Adult

Acute pancreatitis is characterized by an activation cascade of digestive enzymes in the pancreas. The first of these, trypsinogen, can be converted to active trypsin by the peptidase cathepsin B (CTSB). We investigated whether cathepsin L (CTSL) can also process trypsinogen to active trypsin and has a role in pancreatitis.. In CTSL-deficient (Ctsl(-/-)) mice, pancreatitis was induced by injection of cerulein or infusion of taurocholate into the pancreatic duct. Human tissue, pancreatic juice, mouse pancreatitis specimens, and recombinant enzymes were studied by enzyme assay, immunoblot, N-terminal sequencing, immunocytochemistry, and electron microscopy analyses. Isolated acini from Ctsl(-/-) and Ctsb(-/-) mice were studied.. CTSL was expressed in human and mouse pancreas, colocalized with trypsinogen in secretory vesicles and lysosomes, and secreted into pancreatic juice. Severity of pancreatitis was reduced in Ctsl(-/-) mice, whereas apoptosis and intrapancreatic trypsin activity were increased. CTSL-induced cleavage of trypsinogen occurred 3 amino acids toward the C-terminus from the CTSB activation site and resulted in a truncated, inactive form of trypsin and an elongated propeptide (trypsinogen activation peptide [TAP]). This elongated TAP was not detected by enzyme-linked immunosorbent assay (ELISA) but was effectively converted to an immunoreactive form by CTSB. Levels of TAP thus generated by CTSB were not associated with disease severity, although this is what the TAP-ELISA is used to determine in the clinic.. CTSL inactivates trypsinogen and counteracts the ability of CTSB to form active trypsin. In mouse models of pancreatitis, absence of CTSL induces apoptosis and reduces disease severity.

Topics: Amylases; Animals; Apoptosis; Cathepsin B; Cathepsin L; Ceruletide; Disease Models, Animal; Humans; Hydrogen-Ion Concentration; Lipase; Mice; Mice, Knockout; Pancreatitis; Severity of Illness Index; Taurocholic Acid; Trypsin; Trypsinogen

Our aim was to determine if total parenteral nutrition (TPN)-induced pancreatic atrophy and Hsp70 expression attenuates cerulein-induced pancreatitis in rats.. Rats were randomized to a 7-day course of saline infusion plus a semipurified diet or TPN, with or without an intravenous cerulein injection or vehicle on day 7, and killed 1 or 6 hours after the injection. Based on a pilot study, 1 hour was the primary time point. Pancreatic atrophy was determined by mass, protein, and DNA contents. Pancreatic heat shock protein 70 (Hsp70) expression was measured by Western analysis. Histological examination of the pancreas assessed for edema, inflammation, vacuolization, and apoptosis. Serum amylase activity was measured using the Phadebas assay. Pancreatic trypsinogen activation was measured using a fluorometric substrate assay.. The saline-infused rats fed orally gained significantly more weight than TPN rats. The TPN decreased the pancreatic mass and protein content and the protein-DNA ratio and increased the pancreatic DNA content compared with the saline. The TPN increased the pancreatic Hsp70 expression by 91% compared with the saline. The TPN reduced the cerulein-induced pancreatic histological edema, the vacuolization, and the inflammation compared with the saline. The increase in the serum amylase level after cerulein injection was significantly attenuated, and trypsinogen activation was reduced in TPN animals compared with the saline group.. Lack of luminal nutrients with a 7-day course of TPN provides moderate protection against cerulein-induced pancreatitis in rats.

Topics: Amylases; Animals; Ceruletide; HSP70 Heat-Shock Proteins; Male; Pancreatitis; Parenteral Nutrition, Total; Rats; Rats, Sprague-Dawley; Trypsinogen

Autoimmune pancreatitis (AIP) is thought to be an immune-mediated inflammatory process, directed against the epithelial components of the pancreas. The objective was to identify novel markers of disease and to unravel the pathogenesis of AIP.. To explore key targets of the inflammatory process, we analyzed the expression of proteins at the RNA and protein level using genomics and proteomics, immunohistochemistry, western blot, and immunoassay. An animal model of AIP with LP-BM5 murine leukemia virus-infected mice was studied in parallel. RNA microarrays of pancreatic tissue from 12 patients with AIP were compared with those of 8 patients with non-AIP chronic pancreatitis.. Expression profiling showed 272 upregulated genes, including those encoding for immunoglobulins, chemokines and their receptors, and 86 downregulated genes, including those for pancreatic proteases such as three trypsinogen isoforms. Protein profiling showed that the expression of trypsinogens and other pancreatic enzymes was greatly reduced. Immunohistochemistry showed a near-loss of trypsin-positive acinar cells, which was also confirmed by western blotting. The serum of AIP patients contained high titers of autoantibodies against the trypsinogens PRSS1 and PRSS2 but not against PRSS3. In addition, there were autoantibodies against the trypsin inhibitor PSTI (the product of the SPINK1 gene). In the pancreas of AIP animals, we found similar protein patterns and a reduction in trypsinogen.. These data indicate that the immune-mediated process characterizing AIP involves pancreatic acinar cells and their secretory enzymes such as trypsin isoforms. Demonstration of trypsinogen autoantibodies may be helpful for the diagnosis of AIP.

Topics: Adult; Animals; Autoantibodies; Autoimmune Diseases; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Female; Gene Expression Profiling; Humans; Immunoassay; Immunohistochemistry; Inflammation; Logistic Models; Male; Mice; Middle Aged; Oligonucleotide Array Sequence Analysis; Pancreas, Exocrine; Pancreatitis; Proteome; Trypsinogen

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Testing; Humans; Mutation; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The aim of the study was to investigate the role and importance of the urine trypsinogen-2 dipstick test in the differential diagnosis of acute pancreatitis in the Emergency Department and to compare results with those of conventional tests.. The study was performed prospectively in the patients admitting to the Emergency Department due to upper abdominal pain. Thirty-two of the 87 patients included in the study had acute pancreatitis diagnosis. Serum amylase, lipase, C-reactive protein (CRP) and urine trypsinogen-2 using Actim pancreatitis dipstick were studied in all patients. The statistical analysis was performed using SPSS 11.5 package program.. Urine trypsinogen-2 was found positive in 21 (65.6%) of 32 patients. The sensitivity of the test for pancreatitis was identified as 64%, specificity as 85%, positive predictive value as 72%, and negative predictive value as 81%. These values were statistically significant compared to the control group (p<0.01).. Although it has lower sensitivity and specificity compared to amylase and lipase, we suggest that urine trypsinogen-2 test may be an important diagnostic tool in excluding the diagnosis of acute pancreatitis, since it provides results within 5 minutes in the Emergency Department, is cheaper, has a higher negative predictive value, and is easy to use.

Topics: Acute Disease; Amylases; C-Reactive Protein; Diagnosis, Differential; Emergency Service, Hospital; Humans; Lipase; Pain; Pancreatitis; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity; Trypsin; Trypsinogen

The lysosomal protease cathepsin B is thought to play a crucial role in the intracellular activation cascade of digestive proteases and in the initiation of acute pancreatitis. Although cathepsin B has been shown to be physiologically present in the secretory pathway of pancreatic acinar cells it has been suggested that premature activation of zymogens requires an additional redistribution of cathepsin B into the secretory compartment. Here, we studied the role of cathepsin B targeting during caerulein-induced pancreatitis in mouse mutants lacking the cation-independent mannose 6-phosphate/insulin-like growth factor II receptor (CI-MPR) which normally mediates the trafficking of cathepsin B to lysosomes. Absence of the CI-MPR led to redistribution of cathepsin B to the zymogen granule enriched subcellular fraction and to a substantial formation of large cytoplasmic vacuoles that contained both, trypsinogen and cathepsin B. However, this did not cause premature intracellular trypsin activation in saline-treated control animals lacking the CI-MPR. During caerulein-induced pancreatitis, trypsinogen activation in the pancreas of CI-MPR-deficient animals was about 40% higher than in wild-type animals but serum amylase levels were reduced and lung damage was unchanged. These data suggest that subcellular redistribution of cathepsin B, in itself, induces neither spontaneous trypsinogen activation nor pancreatitis. Furthermore, we clearly show that a marked increase in intracellular trypsinogen activation is not necessarily associated with greater disease severity.

Topics: Amylases; Animals; Cathepsin B; Ceruletide; Disease Progression; Insulin-Like Growth Factor II; Lysosomes; Mice; Mice, Knockout; Pancreas; Pancreatitis; Peptide Hydrolases; Receptor, IGF Type 2; Secretory Vesicles; Trypsin; Trypsinogen; Vacuoles

PRSS1 and SPINK1 are 2 important genes in the defense mechanism guarding against the development of pancreatitis. This study aimed to evaluate the prevalence of PRSS1 and SPINK1 mutations and to explore the presence of any ethnic specificity in Korean patients.. A total of 47 patients from 40 families including 37 patients with idiopathic pancreatitis and 10 patients with familial pancreatitis were prospectively enrolled. Fifty healthy controls were included for analysis of SPINK1 IVS3+2T site.. PRSS1 mutations were observed in 6 patients from 2 families and SPINK1 mutations in 13 patients from 11 families, respectively. In case of SPINK1 mutations, N34S and IVS3+2T>C were identified in 3 and 11 patients, respectively, including one with compound N34S/IVS3+2T>C heterozygote. The prevalence of SPINK1 IVS3+2T>C mutations was 26.8% among 41 patients without PRSS1 mutations, whereas the prevalence among 50 healthy controls was 0%. Only PRSS1 R122H was identified. Late onset of symptoms at the age of 36 years and absence of symptoms at the age of 47 years were observed in 2 patients with PRSS1 mutations.. PRSS1 and SPINK1 mutations were not rare in Korean patients with idiopathic and familial pancreatitis. SPINK1 IVS3+2T>C was a prevalent mutation in this population.

Topics: Adolescent; Adult; Carrier Proteins; Child; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Humans; Male; Middle Aged; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

There is a concept that pancreatitis results from an imbalance of proteases and their inhibitors within the pancreatic parenchyma. It has been recently shown that a loss-of-function variant, c.571G>A (p.G191R), in the anionic trypsinogen (PRSS2) gene protects against chronic pancreatitis in European populations. Here we examined the association of the p.G191R variant with pancreatic disorders in Japan.. Genomic DNA was prepared from 378 healthy controls and 604 patients with pancreatic disorders (241 patients with chronic pancreatitis, 174 with acute pancreatitis, and 189 with pancreatic neoplasm). Mutational analysis of the PRSS2 gene was performed by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing.. The heterozygous p.G191R variant was found in three of 241 (1.2%) patients with chronic pancreatitis, in seven of 174 (4.0%) patients with acute pancreatitis, and in 12 of 189 (6.3%) patients with pancreatic neoplasm. The p.G191R variant was found in 25 (two were homozygous and 23 were heterozygous) of 378 (6.6%) healthy controls. The p.G191R frequency in patients with chronic pancreatitis was lower than that in healthy controls (p = 0.001; odds ratio (OR) 0.178; 95% confidence interval (CI) = 0.057 to 0.561). The p.G191R frequency was lower in patients with alcoholic (0.9%; p = 0.015; OR, 0.132; 95% CI, 0.022 to 0.779) and idiopathic (1.0%; p = 0.025; OR, 0.144; 95% CI, 0.025 to 0.851) chronic pancreatitis than that in healthy controls. There were no statistical differences in the p.G191R frequency between healthy controls and patients with acute pancreatitis or with pancreatic neoplasm. Patients with alcoholic acute pancreatitis (n = 59) had no variant carrier, and the p.G191R frequency was lower than that in healthy controls (p = 0.035).. The p.G191R variant protected against alcoholic and idiopathic chronic pancreatitis as well as alcoholic acute pancreatitis in Japan.

Topics: Adenocarcinoma; Adult; Aged; Alcohol Drinking; Case-Control Studies; DNA Mutational Analysis; Female; Gene Frequency; Genotype; Heterozygote; Homozygote; Humans; Japan; Male; Middle Aged; Multivariate Analysis; Mutation; Odds Ratio; Pancreatic Neoplasms; Pancreatitis; Pancreatitis, Acute Necrotizing; Pancreatitis, Chronic; Trypsin; Trypsinogen

We investigated the biochemical properties and cellular expression of the c.346C>T (p.R116C) human cationic trypsinogen (PRSS1) mutant, which we identified in a German family with autosomal dominant hereditary pancreatitis. This mutation leads to an unpaired Cys residue with the potential to interfere with protein folding via incorrect disulfide bond formation. Recombinantly expressed p.R116C trypsinogen exhibited a tendency for misfolding in vitro. Biochemical analysis of the correctly folded, purified p.R116C mutant revealed unchanged activation and degradation characteristics compared to wild type trypsinogen. Secretion of mutant p.R116C from transfected 293T cells was reduced to approximately 20% of wild type. A similar secretion defect was observed with another rare PRSS1 variant, p.C139S, whereas mutants p.A16V, p.N29I, p.N29T, p.E79K, p.R122C, and p.R122H were secreted normally. All mutants were detected in cell extracts at comparable levels but a large portion of mutant p.R116C was present in an insoluble, protease-sensitive form. Consistent with intracellular retention of misfolded trypsinogen, the endoplasmic reticulum (ER) stress markers immunoglobulin-binding protein (BiP) and the spliced form of the X-box binding protein-1 (XBP1s) were elevated in cells expressing mutant p.R116C. The results indicate that mutation-induced misfolding and intracellular retention of human cationic trypsinogen causes hereditary pancreatitis in carriers of the p.R116C mutation. ER stress triggered by trypsinogen misfolding represents a new potential disease mechanism for chronic pancreatitis.

Topics: Adult; Aged; Blotting, Western; Cell Line; Cell Line, Tumor; Child; DNA-Binding Proteins; Endoplasmic Reticulum Chaperone BiP; Family Health; Female; Heat-Shock Proteins; Humans; Male; Middle Aged; Molecular Chaperones; Mutant Proteins; Mutation; Pancreatitis; Pedigree; Protein Folding; Regulatory Factor X Transcription Factors; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors; Trypsin; Trypsinogen; X-Box Binding Protein 1

Topics: Asian People; Calcinosis; Case-Control Studies; Genotype; Humans; India; Mutation; Pancreatitis; Trypsin; Trypsinogen

Hereditary pancreatitis is an autosomal dominant disease which is caused by mutations in the PRSS1 gene.. The aim of our study was to describe the penetrance and phenotype-genotype correlations of the c.346C>T (p.R122C) mutation.. Case series descriptive study.. Forty-one members of six families from whom DNA samples were analyzed.. In subjects with R122C mutation symptoms, pancreatic calcifications, main pancreatic duct changes, diabetes, steatorrhea, pancreatic cancer and surgery were recorded.. The R122C mutation was detected in 22 of the 41 family members studied, and 7 men and 2 women developed pancreatic disease, resulting in a penetrance of 40.9%. One out of the 9 patients was excluded because she died before the mutation was detected. The mean age at symptom onset was 23.5 years (range: 4-51 years). Abdominal pain was present in 6 (75.0%) of the 8 patients with the R122C mutation who developed pancreatic disease. Calcifications had developed in 5 (62.5%) at a mean age of 35.8 years (range: 14-56 years). Five (62.5%) developed changes in the pancreatic ducts at a mean age of 44.2 years (range: 19-65 years). Two patients (25.0%) developed steatorrhea during the follow-up at 26 and 35 years of disease progression. Diabetes developed in five patients (62.5%) at a mean age of 41.4 years old (range: 22-53 years). Three of the patients analyzed (37.5%) developed pancreatic cancer at 59 years of age, 63 years of age and 70 years of age.. Penetrance of the R122C mutation is lower than that described for the R122H and N29I mutations, and there is a tendency toward a predominance of males with the R122C mutation who developed the phenotype of pancreatitis.

Topics: Adult; Aged; Child; Family Health; Female; Genotype; Humans; Male; Middle Aged; Pancreatitis; Pedigree; Penetrance; Phenotype; Point Mutation; Spain; Trypsin; Trypsinogen; Young Adult

Patients with hereditary pancreatitis (HP) bear a high risk of pancreatic adenocarcinoma, but their life expectancy remains unknown. The objective of the study was to assess whether the high risk of cancer decreases survival.. Inclusion criteria were the presence of a PRSS1 mutation with pancreatic symptoms or chronic pancreatitis in at least two first-degree relatives or three second-degree relatives without another cause. Survival rates were assessed according to risk factors. Excess mortality compared with the general French population was calculated (statistical Esteve model) for two periods (20-50 and 50-70 years), according to several risk factors.. The cohort comprised 189 patients. PRSS1 mutations were found in 66%. A total of 19 patients died at the median age of 60. In all, 10 deaths were attributable to HP, including 8 to pancreatic adenocarcinoma. Median overall survival for the whole cohort was 74 years (95% confidence interval (CI): 71-79). The presence of R122H mutation, gender, tobacco consumption in patients older than 18 years, and diabetes mellitus were not associated with differences in survival. Only patients with pancreatic cancer had decreased survival (P=0.008). Excess mortality risk compared with the general population was 0.02% between 20 and 50 years, and 0.61% between 50 and 70 years (NS). Gender, R122H mutation, diabetes, and tobacco use were not associated with excess mortality in these two periods.. Despite their high risk of cancer, HP patients do not have excess mortality risk compared with the general population, irrespective of gender, tobacco use, or diabetes mellitus. These data should be brought to the patient's attention.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; Female; France; Genetic Predisposition to Disease; Humans; Infant; Male; Middle Aged; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsinogen; Young Adult

This study updated the estimated prevalence of type 3c diabetes damage to the pancreas through different genotypes of PRSS1 and their clinical characteristics in the Han population.. Cross-sectional analysis was performed of the most recent (2003-2007) patients with pancreatitis from six hospitals of the Han population in South China (n = 253).. There were 32 patients with pancreatitis carrying a PRSS1 gene abnormality within intron region among 253 cases of pancreatitis, including 27 patients carrying novel single nucleotide polymorphisms, namely, IVS 3 +75 A --> G conversion, and five patients with the mutation IVS3 + 10 T --> G. Among these patients, there were only three cases of patients with diabetes (9.37%). This was lower than the prevalence of abnormalities in the exons of the PRSS1 gene (51.92%): 12 patients with c.361 G --> A, eight patients with c.415 T --> A, and five patients with c.365G --> A. Among them were 12 persons with diabetes, including five requiring insulin to regulate blood sugar. What is more, among the 27 patients carrying PRSS1 gene polymorphism (c.486 C --> T, within the exon 4), there were 15 persons with diabetes symptoms. More than 40% of these patients required insulin to regulate blood sugar.. An abnormality within the intron region of the PRSS1 gene represents one of the causes of pancreatitis in Chinese patients, but it is not related to pancreatic diabetes. However, the exon abnormality obviously raises the morbidity rate of type 3c diabetes, which relies on insulin.

Topics: Adolescent; Adult; Aged; Asian People; Case-Control Studies; Child; Child, Preschool; China; Cross-Sectional Studies; Diabetes Mellitus; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Pancreatitis; Point Mutation; Polymerase Chain Reaction; Polymorphism, Genetic; Prevalence; Trypsin; Trypsinogen; Young Adult

The objective of this study was to investigate the role of MAPKAP kinase 2 (MK2) and heat shock protein (HSP) HSP60 in the pathogenesis of a new model of severe acute pancreatitis (AP). MK2 plays a significant role in the regulation of cytokines. It has been shown that induction and expression of several HSPs can protect against experimental pancreatitis. Interplay between both systems seems of high interest. Mice with a homozygous deletion of the MK2 gene were used. Severe AP was induced by combined intraperitoneal injections of cerulein with lipopolysaccharide (LPS). Severity of AP was assessed by biochemical markers and histology. The serum IL-6 and lung myeloperoxidase (MPO) levels were determined for assessing the extent of systemic inflammatory response. Expression of HSP25, HSP60, HSP70, and HSP90 was analyzed by Western blotting. Repeated injections of cerulein alone or cerulein plus LPS (Cer+LPS) resulted in local inflammatory responses in the pancreas and corresponding systemic inflammatory changes with pronounced severity in the Cer+LPS group. Compared with the C57Bl wild-type mice, the MK2-/- mice presented with significant milder pancreatitis and attenuated responses of serum amylase and trypsinogen activity. Furthermore, serum IL-6 was decreased as well as lung MPO activity. Injection of LPS alone displayed neither pancreatic inflammatory responses nor alterations of pancreatic enzyme activities but evidently elevated serum IL-6 levels and increased lung MPO activity. In contrast hereto, in the MK2-/- mice, these changes were much milder. Increased expression of HSP25 and HSP60 occurred after induction of AP. Especially, HSP60 was robustly elevated after Cer+LPS treatment, in both MK2-/- and wild-type mice. Thus the homozygous deletion of the MK2 gene ameliorates the severity of acute pancreatitis and accompanying systemic inflammatory reactions in a new model of severe acute pancreatitis. Our data support the hypothesis that MK2 participates in the multifactorial regulation of early inflammatory responses in AP, independently of the regulation of stress proteins like HSP25 and HSP60 and most likely due to its effect on cytokine regulation.

Topics: Animals; Ceruletide; Chaperonin 60; Gene Deletion; Heat-Shock Proteins; Interleukin-6; Intracellular Signaling Peptides and Proteins; Lipopolysaccharides; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Molecular Chaperones; Neoplasm Proteins; Pancreas; Pancreatic alpha-Amylases; Pancreatitis; Peroxidase; Protein Serine-Threonine Kinases; Trypsinogen

The aim of this study was to determine whether mutations in SPINK1/PRSS1 genes are associated with benign pancreatic hyperenzymemia (BPH).. Sixty-eight subjects with BPH (including 13 familial cases) were studied. In all, we sequenced germline DNA for all the exons and intro-exon boundaries of PRSS1 and SPINK1.. Nine (13.2%) of the 68 subjects harbored PRSS1 or SPINK1 mutations. As to PRSS1, no hereditary pancreatitis-associated variant was detected, whereas previously undescribed mutations (p.Ala148Val and c.40+1G>A) were respectively found in 2 subjects (2.9%). SPINK1 mutations were detected in 7 subjects (10.3%). Five of them exhibited known mutations (3 p.Asn34Ser, 1 p.Pro55Ser, and 1 c.88-23A>T), whereas 2 had a newly found variant (p.Arg67Gly and c.*32C>T, respectively). Only 2 familial BPH, belonging to 2 different families, were found to carry a mutation (1 with p.Ala148Val for PRSS1 and 1 with p.Asn34Ser for SPINK1).. No known mutations of PRSS1 have been found in BPH, whereas the frequency of known SPINK1 variants is similar to that reported in the general population. No segregation of PRSS1/SPINK1 variants occurs in BPH families. Benign pancreatic hyperenzymemia cannot be explained by mutations in genes whose variants are known to be associated with pancreatitis or by mutations in other PRSS1/SPINK1 genes.

Topics: Adolescent; Adult; Aged; Amylases; Base Sequence; Carrier Proteins; Child; Exons; Female; Genetic Predisposition to Disease; Humans; Introns; Isoamylase; Lipase; Male; Middle Aged; Molecular Sequence Data; Mutation; Pancreas; Pancreatic Diseases; Pancreatitis; Syndrome; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Autophagy is mostly a nonselective bulk degradation system within cells. Recent reports indicate that autophagy can act both as a protector and killer of the cell depending on the stage of the disease or the surrounding cellular environment (for review see Cuervo, A.M. 2004. Trends Cell Biol. 14:70-77). We found that cytoplasmic vacuoles induced in pancreatic acinar cells by experimental pancreatitis were autophagic in origin, as demonstrated by microtubule-associated protein 1 light chain 3 expression and electron microscopy experiments. To analyze the role of macroautophagy in acute pancreatitis, we produced conditional knockout mice lacking the autophagy-related 5 gene in acinar cells. Acute pancreatitis was not observed, except for very mild edema in a restricted area, in conditional knockout mice. Unexpectedly, trypsinogen activation was greatly reduced in the absence of autophagy. These results suggest that autophagy exerts devastating effects in pancreatic acinar cells by activation of trypsinogen to trypsin in the early stage of acute pancreatitis through delivering trypsinogen to the lysosome.

Topics: Animals; Autophagy; Autophagy-Related Protein 5; Ceruletide; Enzyme Activation; Integrases; Mice; Mice, Transgenic; Microtubule-Associated Proteins; Pancreas, Exocrine; Pancreatitis; Trypsin; Trypsinogen

Autodigestion of the pancreas by its own prematurely activated digestive proteases is thought to be an important event in the onset of acute pancreatitis. Although lysosomal hydrolases, such as cathepsin B, play a key role in intrapancreatic trypsinogen activation, it remains unclear where and how trypsinogen meets these lysosomal enzymes. Autophagy is an intracellular bulk degradation system in which cytoplasmic components are directed to the lysosome/vacuole by a membrane-mediated process. To analyze the role of autophagy in acute pancreatitis, we produced a conditional knockout mouse that lacks the autophagy-related (Atg) gene Atg5 in the pancreatic acinar cells. The severity of acute pancreatitis induced by cerulein is greatly reduced in these mice. In addition, Atg5-deficient acinar cells show a significantly decreased level of trypsinogen activation. These data suggest that autophagy exerts a detrimental effect in pancreatic acinar cells by activation of trypsinogen to trypsin. We propose a theory in which autophagy accelerates trypsinogen activation by lysosomal hydrolases under acidic conditions, thus triggering acute pancreatitis in its early stage.

Topics: Acute Disease; Animals; Autophagy; Autophagy-Related Protein 5; Ceruletide; Cytoplasm; Enzyme Activation; Mice; Mice, Knockout; Microtubule-Associated Proteins; Pancreatitis; Trypsinogen; Vacuoles

The pathophysiology of acute pancreatitis (AP) may be studied using markers of protease activation (active carboxypeptidase B (aCAP), the activation peptide of carboxypeptidase B (CAPAP)), leakage of pancreatic enzymes (trypsinogen-2, procarboxypeptidase B (proCAP), amylase), and inflammation (monocyte chemoattractant protein-1 (MCP-1), CRP).. This prospective study included 140 cases of AP. Mild (n = 124) and severe (n = 16) cases were compared with respect to serum levels of trypsinogen-2, proCAP, amylase, aCAP, CAPAP (serum/urine), MCP-1 (serum/urine) and CRP on days 1, 2 and 3 from onset of symptoms. All patients with information on all 3 days were included in a time-course analysis (n = 44-55, except amylase: n = 27).. High levels in severe versus mild cases were seen for trypsinogen-2, CAPAP in serum and urine, and MCP-1 in serum on days 1-3. No differences were seen for proCAP, amylase and aCAP. MCP-1 in urine was significantly elevated on day 1-2, and CRP on day 2-3. CAPAP and MCP-1 levels peaked early and stayed elevated for 48 h in serum.. Protease activation and inflammation are early events in AP, with high levels of these markers within 24 h. Protease activation declines after 48 h, whereas inflammation is present for a longer time.

Topics: Adult; Aged; Aged, 80 and over; Amylases; C-Reactive Protein; Carboxypeptidase B; Chemokine CCL2; Enzyme Activation; Female; Humans; Inflammation; Male; Middle Aged; Pancreatitis; Peptide Hydrolases; Trypsin; Trypsinogen

Heat shock proteins (HSPs), induced by a variety of stresses, are known to protect against cellular injury. Recent studies have demonstrated that prior beta-adrenergic stimulation as well as thermal or culture stress induces HSP70 expression and protects against cerulein-induced pancreatitis. The goal of our current studies was to determine whether or not a non-thermal, chemical stressor like sodium arsenite also upregulates HSP70 expression in the pancreas and prevents secretagogue-induced trypsinogen and NF-kappaB activation. We examined the effects of sodium arsenite preadministration on the parameters of cerulein-induced pancreatitis in rats and then monitored the effects of preincubating pancreatic acini with sodium arsenite in vitro. Our results showed that sodium arsenite pretreatment induced HSP70 expression both in vitro and in vivo and significantly ameliorated the severity of cerulein-induced pancreatitis, as evidenced by the markedly reduced degree of hyperamylasemia, pancreatic edema, and acinar cell necrosis. Sodium arsenite pretreatment not only inhibited trypsinogen activation and the subcellular redistribution of cathepsin B, but also prevented NF-kappaB translocation to the nucleus by inhibiting the IkappaBalpha degradation both in vivo and in vitro. We also examined the effect of sodium arsenite pretreatment in a more severe model of pancreatitis induced by L-arginine and found a similarly protective effect. Based on our observations we conclude that, like thermal stress, chemical stressors such as sodium arsenite also induce HSP70 expression in the pancreas and protect against acute pancreatitis. Thus, non-thermal pharmacologically induced stress can help prevent or treat pancreatitis.

Topics: Actins; Adenosine Triphosphate; Animals; Arginine; Arsenites; Cell Survival; Ceruletide; Dose-Response Relationship, Drug; Enzyme Activation; HSP70 Heat-Shock Proteins; NF-kappa B; Pancreas; Pancreatitis; Protein Transport; Rats; Rats, Wistar; Sodium Compounds; Time Factors; Trypsin; Trypsinogen; Up-Regulation

The mechanism by which hypertriglyceridemia (HTG) leads to pancreatitis is not clear. We sought to determine whether the genes involved in pancreatic ductal or acinar cell injury, including the cationic trypsinogen gene [protease, serine, 1 (trypsin 1) (PRSS1)], the pancreatic secretory trypsin inhibitor gene [serine peptidase inhibitor, Kazal type 1 (SPINK1)], the cystic fibrosis transmembrane conductance regulator gene [cystic fibrosis transmembrane conductance regulator (ATP-binding cassette subfamily C, member 7) (CFTR)], and inflammation genes such as tumor necrosis factor [tumor necrosis factor, TNF superfamily, member 2 (TNF)] are associated with hyperlipidemic pancreatitis (HLP) in patients with HTG.. We performed genetic analysis of 126 HTG patients in Taiwan (46 with HLP and 80 without HLP). The entire coding and intronic regions of the PRSS1, SPINK1, and CFTR genes were identified by heteroduplex analysis techniques and were confirmed by sequencing analysis. The presence of 125G/C, 1001 + 11C>T, 1540A>G (Met470Val), 2694T>G, and 4521G>A in CFTR, the presence of 272C>T in SPINK1, and TNF promoter polymorphisms (nucleotide positions 1031, 863, 857, 308, and 308) were measured by direct sequencing.. Of the 126 HTG patients, 13 (10.3%) carried a CFTR mutation. No PRSS1 or SPINK1 mutations were detected in our patients or in HTG controls. The CFTR gene mutation rates in HTG with and without HLP were 26.1% (12 of 46) and 1.3% (1 of 80), respectively (P <0.0001). The CFTR gene mutations were all Ile556Val. A multivariate analysis of HTG patients indicated that triglycerides, CFTR 470Val, and TNF promoter 863A were independent risk markers for HLP.. This genetic study is the first one to address the association of HLP with the CFTR mutation/variant/haplotype and TNF promoter polymorphism in a Chinese HTG population. The results suggest that the occurrence of HLP is multifactorial and polygenic.

Topics: Adult; Asian People; Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Genetic Variation; Haplotypes; Humans; Hyperlipidemias; Male; Middle Aged; Mutation; Pancreatitis; Polymorphism, Genetic; Promoter Regions, Genetic; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen; Tumor Necrosis Factor-alpha

Primary hyperparathyroidism (pHPT)-related hypercalcemia is considered to represent a risk factor for the development of pancreatitis. We therefore explored whether mutations in genes that were previously identified to increase the risk for pancreatitis coexist in a cohort of 826 patients with pHPT prospectively studied between 1987 and 2002.. Among 826 patients with pHPT, 38 patients were identified with pancreatitis (4.6%). DNA was available from 25 patients (13 women/12 men, 16 acute pancreatitis/9 chronic pancreatitis). These individuals and 50 patients with pHPT without pancreatitis were analyzed for mutations in the serine protease inhibitor Kazal type I (SPINK1) gene (N34S) and the cationic trypsinogen gene (PRSS1) (N29I, R122H) by melting curve analysis and DNA sequencing. Sequence analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene was carried out for the detection of 36 mutations and the Tn polymorphism.. Four of 25 patients with pHPT and pancreatitis carried the N34S missense mutation in the SPINK1 gene (16%), while all 50 controls (pHPT without pancreatitis) showed no mutation in SPINK1 or PRSS1 genes (P < 0.05 vs controls, P < 0.001 vs general population). CF-causing CFTR mutations were present in four patients (P < 0.05 vs general population), while one patient carried a 5T allele. One patient was transheterozygous (SPINK1: N34S/CFTR: R553X). Mean serum calcium levels in pancreatitis patients (3.1 mmol/L) did not differ significantly from the mean of the entire cohort (3.0 mmol/L) or pHPT patients without pancreatitis (3.1 mmol/L).. Pancreatitis risk is approximately 10-fold elevated in pHPT, but pancreatitis occurs infrequently. This indicates an existing but minor impact of pHPT-related hypercalcemia. If pancreatitis occurs, it seems associated with genetic risk factors such as mutations in the SPINK1 and CFTR genes. In contrast, a combination of both hypercalcemia and genetic variants in SPINK1 or CFTR increases the risk to develop pancreatitis in patients with pHPT.

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Humans; Hyperparathyroidism, Primary; Male; Middle Aged; Mutation; Pancreatitis; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

An increased risk of pancreatic adenocarcinoma (PA) in patients with hereditary pancreatitis (HP) was previously demonstrated in two multinational studies. The PA frequency in this setting is however unknown due to lack of exhaustive case collection. The aims of this study were to evaluate the standardized incidence ratio (SIR) of PA in an exhaustive national series of patients with HP and to search for risk factors.. All French genetic laboratories (response rate 100%), pediatricians, and gastroenterologists (response rate 84%) were contacted.. mutation in the PRSS1 gene or recurrent, acute, or chronic pancreatitis, with no precipitating factors in two first-degree relatives or >or=3 second-degree relatives in >or=2 generations. Diagnosis of PA was based on histological records.. Seventy-eight families and 200 patients were included (181 alive, 6,673 person-years, median number of generations 3, men 53%, alcoholism 5%, and smoking 34%). PRSS1 mutations were searched for in 96% of the patients and were detected in 68% (maternal inheritance 54%, R122H 78%, N29I 12%, and others 10%). Ten PA were diagnosed (median age 55 yr). SIR of PA for the whole population, men, and women were 87 (95% CI 42-113), 69 (25-150), and 142 (38-225), respectively, with no influence of genetic mutation. At ages 50 and 75 yr, the cumulated risk of PA was 11% and 49% for men and 8% and 55% for women, respectively. Smoking and diabetes mellitus were the main associated risk factors.. Patients with HP have a marked relative and absolute increased risk of PA as compared to the general population, especially in smokers. There is no correlation with the type of PRSS1 mutation.

Topics: Adenocarcinoma; Adolescent; Adult; Aged; Aged, 80 and over; Child; Child, Preschool; DNA; Female; France; Genetic Predisposition to Disease; Humans; Incidence; Infant; Male; Middle Aged; Mutation; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

Mutations in the cationic trypsinogen gene (PRSS1) have been detected in patients with hereditary pancreatitis (HP). This study investigated the prevalence of the R122H (c.365 G > A), A121T (c.361 G > A) and D162D (c.488 C > T) mutations or polymorphisms in the common, non-hereditary forms of chronic pancreatitis and in an HP family.. DNA was prepared from blood samples of 54 patients with chronic pancreatitis (35 alcoholic, 17 idiopathic and 2 hereditary) and 120 normal controls. The PRSS1 genes were amplified by polymerase chain reaction (PCR) and their products were analyzed by sequencing and related clinical data were also collected.. A new polymorphism (c.488 C > T) of PRSS1 was found in 25 patients with chronic pancreatitis (including one affected member of the HP family) and six members of the normal controls. The C/T genotype was significantly increased in chronic pancreatitis (OR: 16.379, 95% CI: 5.7522 - 52.3663), the frequency of c.488 C > T change was in according with the Hardy-Weinberg equilibrium, but it doesn't affect the clinical phenotype. The commonly reported change of R122H (c.365 G > A) was not detected in any of the study subjects. c.361 G > A was found in 2 affected members and one unaffected carrier in an HP family. One of the affected members of an HP family had c.361 G > A mutation and polymorphism (c.488 C > T) in the PRSS1 gene at the same time. The patient's clinical values (C3, C4, CA19-9 and HbA1c) were higher than those of the other patients with chronic pancreatitis. The two patients with HP developed diabetes mellitus and their father died with pancreatic cancer.. A new polymorphism (c.488 C > T) in the PRSS1 gene is associated with chronic pancreatitis, but it did not affect the clinical phenotype while the A121T (c.361 G > A) mutation in the gene shows a significant correlation in the patients with HP.

Topics: Female; Humans; Male; Mutation; Pancreatitis; Pancreatitis, Chronic; Polymorphism, Genetic; Trypsin; Trypsinogen

Trypsinogen activation and inflammation are early events in acute pancreatitis. This experimental study aimed to show the effects of dexamethasone on them.. Cerulein and taurocholate pancreatitis were induced in 2 groups of 12 Wistar rats each. Six animals per group were injected with dexamethasone 1 h prior to the induction of acute pancreatitis. Amylase, phospholipase A2, TNF-alpha, IL-6, IL-10, alpha2-antiplasmin in plasma and trypsinogen activation peptide (TAP) in urine were measured in healthy rats, then 0.5 and 6 h after pancreatitis induction. A severity score based on edema, necrosis and ascites was calculated at 6 h. TNF-alpha, IL-6 and IL-10 were measured 0.5 h after laparotomy in a control sham-operated group of 6 rats.. Inflammatory markers increased early in the course of both mild and severe acute pancreatitis and were significantly lowered by dexamethasone. The severity score was higher in taurocholate than in cerulein pancreatitis. It was significantly decreased by dexamethasone only in rats with mild pancreatitis. TAP remained unchanged in mild pancreatitis compared to healthy animals but increased late in the course of taurocholate pancreatitis. Trypsinogen activation was not affected by dexamethasone at all.. Inflammation occurred earlier than the increase in urinary TAP in severe pancreatitis in rats. Dexamethasone inhibited inflammation but had no influence on TAP levels in experimental mild and severe acute pancreatitis.

Topics: Animals; Anti-Inflammatory Agents; Ceruletide; Dexamethasone; Female; Oligopeptides; Pancreatitis; Rats; Rats, Wistar; Taurocholic Acid; Trypsinogen

Leukocyte infiltration is an early and critical event in the development of acute pancreatitis. However, the mechanism of leukocyte transmigration into the pancreas and the function of leukocytes in initiating acute pancreatitis are still poorly understood. Here, we studied the role of S100A9 (MRP14), a calcium binding protein specifically released by polymorph nuclear leukocytes (PMN), in the course of acute experimental pancreatitis. Acute pancreatitis was induced by repeated supramaximal caerulein injections in S100A9 deficient or S100A9 wild-type mice. We then determined S100A9 expression, trypsinogen activation peptide (TAP) levels, serum amylase and lipase activities, and tissue myeloperoxidase (MPO) activity. Cell-cell contact dissociation was analyzed in vitro with biovolume measurements of isolated acini after incubation with purified S100A8/A9 heterodimers, and in vivo as measurement of Evans Blue extravasation after intravenous application of S100A8/A9. Pancreatitis induced increased levels of S100A9 in the pancreas. However, infiltration of leukocytes and MPO activity in the lungs and pancreas during acute pancreatitis was decreased in S100A9-deficient mice and associated with significantly lower serum amylase and lipase activities as well as reduced intrapancreatic TAP-levels. Incubation of isolated pancreatic acini with purified S100A8/A9-heterodimers resulted in a rapid dissociation of acinar cell-cell contacts which was highly calcium-dependent. Consistent with these findings, in vivo application of S100A8/A9 in mice was in itself sufficient to induce pancreatic cell-cell contract dissociation as indicated by Evans Blue extravasation. These data show that the degree of intrapancreatic trypsinogen activation is influenced by the extent of leukocyte infiltration into the pancreas which, in turn, depends on the presence of S100A9 that is secreted from PMN. S100A9 directly affects leukocyte tissue invasion and mediates cell contact dissociation via its calcium binding properties.

Topics: Animals; Biomarkers; Calcium; Calgranulin A; Calgranulin B; Ceruletide; Cholecystokinin; Enzyme Activation; Humans; Intercellular Junctions; Leukocytes; Lung; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Pancreas; Pancreatitis; S100 Proteins; Trypsinogen

Hereditary pancreatitis, an autosomal dominant disease with approximately 80% penetrance, can be caused by both 'gain-of-function' missense and copy number mutations in the cationic trypsinogen gene (PRSS1). Here we demonstrate a heterozygous hybrid PRSS2 (encoding anionic trypsinogen)/PRSS1 gene in a French white family with hereditary pancreatitis, by means of quantitative fluorescent multiplex PCR and RT-PCR analyses. The hybrid gene, in which exons 1 and 2 are derived from PRSS2 and exons 3-5 from PRSS1, apparently resulted from a non-allelic homologous recombination (NAHR) event between the chromosome 7 homologs or sister chromatids during meiosis. Interestingly, this hybrid gene causes the disease through a combination of its inherent 'double gain-of-function' effect, acting simultaneously as a 'quantitative' copy number mutation and a 'qualitative' missense mutation (i.e. the known disease-causing p.N29I mutation). Our finding reveals a previously unknown mechanism causing human inherited disease, enriches the lexicon of human genetic variation and goes beyond the known interaction between copy number variations (CNVs) and single nucleotide substitutions in health and disease. Our finding should also stimulate more interest in analyzing both types of genetic variation whenever one tries to determine the contribution of a specific locus to a given disease phenotype.

Topics: Amino Acid Sequence; Amino Acid Substitution; Base Sequence; Chronic Disease; Female; Genetic Diseases, Inborn; Humans; Male; Molecular Sequence Data; Mutant Chimeric Proteins; Mutation, Missense; Pancreatitis; Pedigree; Trypsin; Trypsinogen

The urine trypsinogen strip test has been used successfully in the diagnosis of pancreatitis of various aetiologies, but has not been studied in postoperative pancreatitis. The aim of this study was to evaluate this test for the diagnosis of postoperative pancreatitis after pancreatic resection.. Fifty patients undergoing pancreatic resection were included. The urine trypsinogen strip test was done daily during the first postoperative week, blood was analysed before and 4, 6 and 10 days after surgery, and amylase activity in the drainage fluid was measured on days 4 and 6. Patients underwent computed tomography (CT) before operation and on days 2 and 6 afterwards.. Thirteen patients (26 per cent) developed CT-detected pancreatitis after operation. In 12 of these patients pancreatitis was detected on the second postoperative day. The urine trypsinogen test was positive in all 13 patients with postoperative pancreatitis, and was already positive on the first day after surgery in 12. The sensitivity, specificity, and positive and negative predictive values of the trypsinogen strip test in detection of postoperative pancreatitis were 100, 92, 81 and 100 per cent respectively. In receiver-operator characteristic analysis the area under the curve (AUC) was higher for the urine trypsinogen strip test (AUC 0.959) than for a serum amylase level more than two (AUC 0.731) or three times (AUC 0.654) above the upper normal range in the diagnosis of postoperative pancreatitis. Patients whose recovery was complicated by pancreatic fistula, detected by drain output measurements on day 6, more often had a positive urine trypsinogen test than patients without a fistula (11 of 12 versus five of 38; P < 0.001).. This study suggests that the urine trypsinogen strip test might be a valuable method for diagnosis of pancreatitis after pancreatic surgery.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers; Child; Female; Humans; Male; Middle Aged; Pancreatectomy; Pancreatitis; Postoperative Complications; Predictive Value of Tests; Reagent Strips; Sensitivity and Specificity; Trypsinogen

To assess a point-of-care (POC) urine trypsinogen (UT) test for the diagnosis of pancreatitis in the emergency department (ED).. This was a prospective cohort study of a convenience sample of patients presenting to the ED with abdominal pain or symptoms suggestive of pancreatitis. A 3-minute POC UT test (Actim Pancreatitis; Medix Biochemica, Kauniainen, Finland) was compared with plasma lipase and amylase measurements, imaging results when performed, and final discharge diagnoses. The criterion standard was a final discharge diagnosis of acute pancreatitis.. Of 191 patients included in this study, 17 patients were diagnosed with either acute or acute-on-chronic pancreatitis. The sensitivity and specificity of UT for acute pancreatitis were, respectively, 100% (95% confidence interval [CI] = 77% to 100%) and 96% (95% CI = 92% to 98%). Seven of the 17 patients with pancreatitis (41%) had diagnostic findings on CT and positive UT tests but had nondiagnostic plasma lipase and amylase levels.. A POC UT screening test for pancreatitis in the ED compared favorably with plasma lipase and amylase levels. Future studies should be performed to explore whether this test in the ED setting has better clinical utility than plasma lipase or amylase.

Topics: Abdominal Pain; Acute Disease; Amylases; Humans; Lipase; Pancreatitis; Pancreatitis, Chronic; Point-of-Care Systems; Prospective Studies; Sensitivity and Specificity; Trypsinogen

In contrast to supramaximal CCK-8 or caerulein, acute or prolonged supraphysiological levels of endogenous CCK-58 do not cause pancreatitis. Compared with CCK-8, CCK-58 is a much stronger stimulant of pancreatic chloride and water secretion, equivalent to maximally effective secretin, but with a chloride-to-bicarbonate ratio characteristic of acinar fluid. Because supraphysiological endogenous CCK does not cause pancreatitis and because coadministration of secretin ameliorated caerulein- or CCK-8-induced pancreatitis, coincident with restoring pancreatic water secretion, we hypothesized that supramaximal CCK-58 would not induce pancreatitis. Conscious rats were infused intravenously with 2 or 4 nmol x kg(-1) x h(-1) of CCK-8 or synthetic rat CCK-58 for 6 h, and pancreases were examined for morphological and biochemical indexes of acute pancreatitis. A second group was treated as above while monitoring pancreatic protein and water secretion. CCK-8 at 2 nmol x kg(-1) x h(-1) caused severe edematous pancreatitis as evidenced by morphological and biochemical criteria. CCK-58 at this dose had minimal or no effect on these indexes. CCK-58 at 4 nmol x kg(-1) x h(-1) increased some indexes of pancreatic damage but less than either the 2 or 4 nmol x kg(-1) x h(-1) dose of CCK-8. Pancreatic water and protein secretion were nearly or completely abolished within 3 h of onset of CCK-8 infusion, whereas water and protein secretion were maintained near basal levels in CCK-58-treated rats. We hypothesize that supramaximal CCK-58 does not induce pancreatitis because it maintains pancreatic acinar chloride and water secretion, which are essential for exocytosis of activated zymogens. We conclude that CCK-58 may be a valuable tool for investigating events that trigger pancreatitis.

Topics: Amylases; Animals; Body Water; Chlorides; Cholecystokinin; Dose-Response Relationship, Drug; Edema; Interleukin-6; Male; Organ Size; Pancreas; Pancreatic Juice; Pancreatitis; Peroxidase; Rats; Rats, Wistar; Secretory Rate; Sincalide; Time Factors; Trypsin; Trypsinogen

Topics: Acute Disease; Animals; Cytokines; Disease Models, Animal; Humans; I-kappa B Kinase; NF-kappa B; Pancreas; Pancreatitis; Trypsinogen

We have hypothesized that the colocalization of digestive zymogens with lysosomal hydrolases, which occurs during the early stages of every experimental pancreatitis model, facilitates activation of those zymogens by lysosomal hydrolases such as cathepsin B and that this activation triggers acute pancreatitis by leading to acinar cell injury. Some, however, have argued that the colocalization phenomenon may be the result, rather than the cause, of zymogen activation during pancreatitis. To resolve this controversy and explore the causal relationships between zymogen activation and other early pancreatitis events, we induced pancreatitis in mice by repeated supramaximal secretagogue stimulation with caerulein. Some animals were pretreated with the cathepsin B inhibitor CA-074 me to inhibit cathepsin B, prevent intrapancreatic activation of digestive zymogens, and reduce the severity of pancreatitis. We show that inhibition of cathepsin B by pretreatment with CA-074 me prevents intrapancreatic zymogen activation and reduces organellar fragility, but it does not alter the caerulein-induced colocalization phenomenon or subcellular F-actin redistribution or prevent caerulein-induced activation of NF-kappaB, ERK1/2, and JNK or upregulated expression of cytochemokines. We conclude 1) that the colocalization phenomenon, F-actin redistribution, activation of proinflammatory transcription factors, and upregulated expression of cytochemokines are not the results of zymogen activation, and 2) that these early events in pancreatitis are not dependent on cathepsin B activity. In contrast, zymogen activation and increased subcellular organellar fragility during caerulein-induced pancreatitis are dependent on cathepsin B activity.

Topics: Actins; Acute Disease; Amylases; Animals; Arylsulfatases; Cathepsin B; Ceruletide; Chemokine CCL2; Dipeptides; Disease Models, Animal; Enzyme Activation; Enzyme Inhibitors; Interleukin-6; JNK Mitogen-Activated Protein Kinases; Lysosomes; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; NF-kappa B; Pancreas; Pancreatitis; Protein Transport; Secretory Vesicles; Severity of Illness Index; Time Factors; Trypsin; Trypsinogen

This study aimed to prove that the urinary trypsinogen-2 dip stick test can be used for early diagnosis of pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).. For this prospective, blinded, pilot study, urine samples were collected before ERCP, 1 h after ERCP, and 4 h after ERCP. The urine dipstick test was used to detect trypsinogen-2 on the basis of immunochromatography. The dipstick results were compared with those of current methods used to diagnose post-ERCP pancreatitis. Once the patient disposition was finalized, pancreatic enzymes, clinical findings, and final diagnosis were obtained from the chart and compared with the urine trypsinogen-2 test findings. The sensitivity, specificity, and positive and negative predictive values were calculated.. The urine trypsinogen dip stick test was performed for 30 patients (15 men and 15 women). Post-ERCP pancreatitis was diagnosed in 5 of 29 patients by clinician assessment, serum pancreatic enzyme levels, or both. The amylase and lipase levels for post-ERCP patients with and without pancreatitis were 650 +/- 145 vs 134 +/- 26 (p = 0.023) and 1,658 +/- 594 vs 84 +/- 17 (p = 0.057), respectively. This statement proves that patients who developed post ERCP pancreatitis had significant elevation of amylase and lipase compared to patients who did not have pancreatitis. For the dip stick test, 6 of 28 patients had positive results in 1 h and 6 of 29 patients had positive results in 4 h. The sensitivity of the 1-h test was 1.0, and the specificity was 0.91. The positive predictive value (PPV) was 0.66, and the negative predictive value (NPV) was 1.0. The sensitivity of the 4-h test was 1.0, and the specificity was 0.96. The PPV was 0.8, and NPV value was 1.0.. The urinary trypsinogen-2 dip stick test is useful for early diagnosis of post-ERCP pancreatitis and allows the testing physicians to begin management early in its course.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Cholangiopancreatography, Endoscopic Retrograde; Early Diagnosis; Female; Humans; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Reagent Strips; Sensitivity and Specificity; Trypsin; Trypsinogen

The intracellular activation of trypsinogen, which is both pH- and calcium-dependent, is an important early step in the development of acute pancreatitis. The cellular compartment in which trypsinogen activation occurs currently is unknown. We therefore investigated the site of intracellular trypsinogen activation by using an established cellular model of acute pancreatitis: supramaximal stimulation of pancreatic acinar cells with cholecystokinin. We used fluorescent dextrans as fluid phase tracers and observed the cholecystokinin-elicited formation and translocation of large endocytic vacuoles. The fluorescent probe rhodamine 110 bis-(CBZ-L-isoleucyl-L-prolyl-L-arginine amide) dihydrochloride (BZiPAR) was used to detect trypsinogen activation. Fluid phase tracers were colocalized with cleaved BZiPAR, indicating that trypsinogen activation occurred within endocytic vacuoles. The development of BZiPAR fluorescence was inhibited by the trypsin inhibitor benzamidine. Fluorescein dextran and Oregon Green 488 BAPTA-5N were used to measure endosomal pH and calcium, respectively. The pH in endocytic vacuoles was 5.9 +/- 0.1, and the calcium ion concentration was 37 +/- 11 microM. The caged calcium probe o-nitrophenyl EGTA and UV uncaging were used to increase calcium in endocytic vacuoles. This increase of calcium caused by calcium uncaging was followed by recovery to the prestimulated level within approximately 100 s. We propose that the initiation of acute pancreatitis depends on endocytic vacuole formation and trypsinogen activation in this compartment.

Topics: Animals; Calcium; Cells, Cultured; Dextrans; Endocytosis; Enzyme Activation; Fluorescent Dyes; Hydrogen-Ion Concentration; Mice; Pancreas; Pancreatitis; Protein Transport; Trypsin; Trypsinogen; Vacuoles

Acute pancreatitis is an autodigestive disease, in which the pancreatic tissue is damaged by the digestive enzymes produced by the acinar cells. Among the tissues in the mammalian body, pancreas has the highest concentration of the natural polyamine, spermidine. We have found that pancreas is very sensitive to acute decreases in the concentrations of the higher polyamines, spermidine and spermine. Activation of polyamine catabolism in transgenic rats overexpressing SSAT (spermidine/spermine-N(1)-acetyltransferase) in the pancreas leads to rapid depletion of these polyamines and to acute necrotizing pancreatitis. Replacement of the natural polyamines with methylated polyamine analogues before the induction of acute pancreatitis prevents the development of the disease. As premature trypsinogen activation is a common, early event leading to tissue injury in acute pancreatitis in human and in experimental animal models, we studied its role in polyamine catabolism-induced pancreatitis. Cathepsin B, a lysosomal hydrolase mediating trypsinogen activation, was activated just 2 h after induction of SSAT. Pre-treatment of the rats with bismethylspermine prevented pancreatic cathepsin B activation. Analysis of tissue ultrastructure by transmission electron microscopy revealed early dilatation of rough endoplasmic reticulum, probable disturbance of zymogen packaging, appearance of autophagosomes and later disruption of intracellular membranes and organelles. Based on these results, we suggest that rapid eradication of polyamines from cellular structures leads to premature zymogen activation and autodigestion of acinar cells.

Topics: Acute Disease; Animals; Disease Models, Animal; Enzyme Activation; Humans; Pancreas; Pancreatitis; Polyamines; Trypsinogen

We developed a one-step immunochromatography assay kit to measure high levels of canine trypsin-like immunoreactivity (cTLI) for bedside estimation of canine pancreatitis. The serum cTLI level can be determined within 10 min by visual comparison of color strengths in the test and reference zones. The serum cTLI levels determined by this method correlate well with canine TLI-ELISA and can be classified into 3 categories: cTLI levels higher than 60 ng/ml were considered positive; 20-60 ng/ml, weakly positive; and less than 20 ng/ml, negative. Twelve dogs suspected of pancreatitis were examined using this method; 4 dogs were positive, 2 were weakly positive, and 6 were negative. This test can detect a high level of serum cTLI and a positive result in the TLIH test will provide critical information for evaluation of pancreatitis in dogs.

Topics: Animals; Dog Diseases; Dogs; Female; Immunoassay; Male; Pancreatitis; Sensitivity and Specificity; Trypsin; Trypsinogen

Mutations in the cationic trypsinogen gene (PRSS1) have been linked with hereditary pancreatitis (HP). A change in R122H in the third exon is one of the mutations most frequently associated with HP. A mutation N34S in the serine protease inhibitor Kazal type 1 gene has also been shown to be linked with HP. The purpose of this study was to report on the incidence of PRSS1 and SPINK1 mutations in a Finnish family with HP and to correlate the findings to the clinical symptoms.. The study included 36 individuals from one Finnish family with HP (21 M, 15 F, median age 38 years). All individuals underwent abdominal ultrasound and laboratory tests (glucose, faecal elastase-1 test). Blood samples were taken for mutational analysis of PRSS1 (R122H, N29I and A16V) and SPINK1 (N34S).. Ten (28%) individuals were affected by mutations: the most frequent mutation was R122H, affecting 8 (22%) individuals; 2 (6%) individuals were affected by the N34S mutation and none by the other tested mutations (N29I and A16V). Four out of eight (50%) R122H-positive individuals had a diagnosis of chronic pancreatitis without other known aetiologies. Four out of five (80%) male individuals with the R122H mutation also had clinical pancreatitis, whereas none of the three mutation-positive females had any signs or symptoms of chronic pancreatitis. The two individuals with the N34S mutation did not have any signs of chronic pancreatitis.. In the investigated Finnish pedigree with HP, the PRSS1 mutation R122H is linked with chronic disease. Although the SPINK1 mutation (N34S) was also observed in two individuals, it was not linked with the disease.

Topics: Adult; Aged, 80 and over; Carrier Proteins; Female; Finland; Genetic Testing; Humans; Male; Middle Aged; Mutation; Pancreatitis; Pedigree; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

We report the identification of a novel mutation in the protease serine 1 (PRSS1) gene in a hereditary pancreatitis (HP) family. Relevant clinical data were collected in this 24-member family. Moreover, the PRSS1 gene was amplified from the genomic DNA of the members with pancreatitis. The amplified products were analyzed by sequencing. A cytosine (C) to thymine (T) mutation in PRSS1 exon3 was detected in four affected members. This novel PRSS1 mutation may be an important factor associated with HP.

Topics: Adult; Asian People; Base Sequence; DNA Mutational Analysis; Exons; Female; Humans; Male; Mutation; Pancreatitis; Pedigree; Polymerase Chain Reaction; Trypsin; Trypsinogen

Genetic predispositions play a major role in the development of chronic pancreatitis. Recently, a mutation in the keratin 8 gene (G62C) was reported to be associated with chronic pancreatitis in Italy. We determined whether mutations in the keratin 8 gene are associated with familial, sporadic and alcoholic recurrent acute or chronic pancreatitis in a population from the United States.. We investigated the relevant genomic region of the keratin 8 gene in 80 patients with familial pancreatitis without a cationic trypsinogen (PRSS1) gene mutation from 52 different families, 21 patients with familial hereditary pancreatitis and a PRSS1 mutation from 20 different families, 126 patients with sporadic pancreatitis without a PRSS1 mutation, 61 patients with alcoholic pancreatitis and 271 controls by direct DNA sequencing.. We found the heterozygous G62C mutation in n = 3/80 patients (n = 2/52 patients from different families, 3.8%) with familial pancreatitis without PRSS1 mutation and in n = 3/126 patients (2.4%) with sporadic pancreatitis. We detected an adjacent heterozygous I63V mutation in n = 2/80 patients (n = 2/52 patients from different families, 3.8%) with familial pancreatitis without PRSS1 mutation and in n = 1/61 patients (1.6%) with alcoholic pancreatitis. We found the G62C mutation in n = 2/271 controls (0.7%) and the I63V mutation in n = 2/271 controls (0.7%). There were no statistically significant differences in the genotype frequencies between patients and controls (p > 0.05). Screening of additional available family members revealed that these variants did not segregate with the disease phenotype. There was no statistically significant difference in the frequency of these keratin 8 variants between patients with chronic pancreatitis and controls (p > 0.05).. These keratin 8 variants are not associated with familial, sporadic or alcoholic pancreatitis.

Topics: Adult; Codon; DNA; Female; Humans; Keratins; Male; Mutation; Pancreatitis; Pancreatitis, Alcoholic; Phenotype; Trypsin; Trypsinogen; United States

The cationic trypsinogen (PRSS1) R122H mutation causes autosomal dominant hereditary pancreatitis (HP) with multiple attacks of acute pancreatitis, but the penetrance, frequency, and severity of attacks are highly variable. HP twins study suggests that modifier genes influence severity but not penetrance.. To investigate potential trypsin associated factors in subjects with the PRSS1 R122H mutation and phenotypic non-penetrance.. Two subjects from HP families (including a 93 year old subject with PRSS1 R122H without pancreatitis), one with chronic pancreatitis and one with a normal pancreas, were studied. Relative expression of: (a) the PRSS1 R122 and H122 alleles; and (b) the PRSS1 and SPINK1 genes in pancreatitis were determined using complementary methods.. PRSS1 wild-type (R122) and mutant (H122) allele expression was equivalent in multiple (> 3) samples from the phenotypically affected and non-penetrant subjects with R122H genotypes using allele specific quantitative reverse transcription-polymerase chain reaction (RT-PCR) and intron spanning nested RT-PCR followed by cDNA sequencing. Compared with PRSS1 mRNA levels, SPINK1 mRNA levels were low in normal appearing tissue but markedly increased in samples with chronic inflammation, independent of PRSS1 genotype.. Attacks of acute pancreatitis in HP subjects appear to be independent of the relative expression of the mutant PRSS1 H122 allele or SPINK1 gene expression. The marked increase in SPINK1 gene expression with inflammation is consistent with its regulation as an acute phase protein.

Topics: Aged, 80 and over; Alleles; Base Sequence; Carrier Proteins; Case-Control Studies; DNA, Complementary; Exons; Gene Expression; Genetic Predisposition to Disease; Humans; Male; Molecular Sequence Data; Mutation; Pancreas; Pancreatitis; Penetrance; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Hereditary pancreatitis is an uncommon autosomal dominant disease secondary to a mutation normally located in the trypsinogen gene, preventing trypsin deactivation. This mutation translates clinically into recurrent attacks of acute pancreatitis and an increased risk of pancreatic cancer. We report a case of acute hereditary pancreatitis due to a trypsinogen mutation that has previously been described in only one family.

Topics: Child; Humans; Male; Mutation; Pancreatitis; Pedigree; Trypsinogen

Defects of PRSS1, SPINK1, CFTR and AAT are considered causative or predisposing to pancreatitis. The aim of this study was to evaluate the impact of these defects into molecular pathology of chronic pancreatitis (CP) and acute recurrent pancreatitis (ARP).. Ninety-two children with CP or ARP, 55 family members and 50 controls were investigated. The subjects were screened for PRSS1 mutations: R122H, R122C, A16V, N29I; SPINK1 N34S variant; panel of 14 CFTR defects: INNOLiPA CFTR12, CFTRdele2,3 and IVS8-T variant or panel of 3 CFTR defects-F508del, CFTRdele2,3 and IVS8-T; AAT mutations: E264V, E342K.. We identified 1 mutated allele in at least 1 of 4 genes in 31 of 92 patients and 12 of 50 controls (P = 0.157). Mutations in SPINK1 and PRSS1 were most frequent. PRSS1 mutations were identified mainly in CP patients (9.6% of CP vs 2.5% of ARP alleles, P = 0.094), whereas N34S SPINK1 mutation was present with comparable frequency in CP and ARP patients (7.7% vs 10.0%, P = 0.768). The frequency of mutations in CFTR alleles was similar to controls (4.9% vs 5%, P = 0.587). Overall frequency of AAT mutations was lower than in the controls. Family studies showed that defects in the examined genes did not always segregate with disease.. PRSS1 defects seem to be causative for pancreatitis, whereas defects in SPINK1 are suggested to be associated with the disease. No association between CFTR mutations and pancreatitis was observed. The importance of AAT variants remains speculative.

Topics: Acute Disease; Adolescent; Adult; Alleles; alpha 1-Antitrypsin; Carrier Proteins; Child; Child, Preschool; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; DNA; DNA Mutational Analysis; Female; Gene Frequency; Genetic Predisposition to Disease; Humans; Male; Mutation; Pancreatitis; Recurrence; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Chronic pancreatitis is a progressive inflammatory disorder leading to irreversible exocrine and/or endocrine impairment. It is well documented that mutations in the cationic trypsinogen (PRSS1) gene can cause hereditary pancreatitis. Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) and the serine protease inhibitor Kazal type 1 (SPINK1) genes are also associated with pancreatitis.. We analyzed 381 patients with a primary diagnosis of chronic or recurrent pancreatitis using the Ambry Test: Pancreatitis to obtain comprehensive genetic information for the CFTR, SPINK1, and PRSS1 genes.. The results identified 32% (122/381) of patients with 166 mutant CFTR alleles, including 12 novel CFTR variants: 4375-20 A>G, F575Y, K598E, L1260P, G194R, F834L, S573C, 2789 + 17 C>T, 621+83 A>G, T164S, 621+25 A>G, and 3500-19 G>A. Of 122 patients with CFTR mutations, 5.5% (21/381) also carried a SPINK1 mutation, and 1.8% (7/381) carried a PRSS1 mutation. In addition, 8.9% (34/381) of all patients had 1 of 11 different SPINK1 mutations. Another 6.3% (24/381) of the patients had 1 of 8 different PRSS1 mutations. Moreover, 1.3% of the patients (5/381) had 1 PRSS1 and 1 SPINK1 mutation. A total 49% (185/381) of the patients carried one or more mutations.. Comprehensive testing of the CFTR, PRSS1, and SPINK1 genes identified genetic variants in nearly half of all subjects considered by their physicians as candidates for genetic testing. Comprehensive test identified numerous novel variants that would not be identified by standard clinical screening panels.

Topics: Acute Disease; Adolescent; Adult; Aged; Carrier Proteins; Child; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Genetic Variation; Humans; Infant; Male; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The R122H mutation of the cationic trypsinogen was found in patients with hereditary pancreatitis. A transgenic animal carrying this mutation could be useful as a genetic model system of pancreatitis.. Mice transgenic for the human R122H cationic trypsinogen were generated using the -205 fragment of the rat elastase promoter. The presence of the transgene was assayed in the DNA, in pancreatic mRNA and in zymogen granule lysates. Serum levels of amylase, lipase and cytokines (MCP-1, IL-6) were monitored and the histological appearance of the tissue was investigated. Pancreatitis was induced by 7 hourly injections of 50 mug/kg cerulein. The procedure was repeated twice weekly for 10 consecutive weeks. The animals were sacrificed 24 (n = 8) and 48 hours (n = 8) after the first injection and at the end of the whole treatment (n = 7).. The transgene was detected at the genomic level and in pancreatic mRNA. The corresponding protein was found in low amounts in zymogen granule lysates. R122H mice showed elevated pancreatic lipase, but there was no spontaneous development of pancreatitis within 18 months. After induction of pancreatitis, levels of lipase (after 24 hours) and amylase (after 48 hours) were higher in R122H mice compared to controls. Repeated treatment with cerulein resulted in a slightly more severe pancreatitis in R122H animals. Amylase, lipase, and the cytokine levels were similar to controls.. The R122H transgenic mouse failed to develop a spontaneous pancreatitis but a repeatedly provoked cerulein-induced pancreatitis led to a slightly more severe pancreatitis. The rather small difference in comparison to controls could be due to the low expression of the transgene in the mouse pancreas.

Topics: Amylases; Animals; Arginine; Ceruletide; Cytokines; Disease Models, Animal; DNA; Histidine; Humans; Lipase; Mice; Mice, Transgenic; Mutation; Pancreas; Pancreatitis; Phenotype; RNA, Messenger; Secretory Vesicles; Severity of Illness Index; Time Factors; Transgenes; Trypsin; Trypsinogen

Hereditary pancreatitis has been reported to be caused by 'gain-of-function' missense mutations in the cationic trypsinogen gene (PRSS1). Here we report the triplication of a approximately 605-kb segment containing the PRSS1 gene on chromosome 7 in five families with hereditary pancreatitis. This triplication, which seems to result in a gain of trypsin through a gene dosage effect, represents a previously unknown molecular mechanism causing hereditary pancreatitis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Chromosomes, Human, Pair 7; Female; Gene Dosage; Gene Duplication; Humans; In Situ Hybridization, Fluorescence; Male; Pancreatitis; Pedigree; Trypsin; Trypsinogen

Little is known about the molecular and biological aspects of the epidemiological association between smoking and pancreatic pathology, such as chronic pancreatitis and pancreatic cancer. Recently, we reported that tobacco smoke exposure induced morphological alterations in the rat pancreas. Here, we have investigated the alterations in the expression of genes associated with exocrine pancreatic function and cellular differentiation upon exposure to cigarette smoke.. Female rats were exposed to environmental smoke inhalation for 2 d/wk (70 min/d) for 12 weeks. The expression profiles of trypsinogen, pancreas-specific trypsin inhibitor, cholecystokinin A receptor, cystic fibrosis transmembrane conductance regulator (CFTR), carbonic anhydrase, and Muc1 and Muc4 mucins transcripts were analyzed by RNA slot blot analysis. Muc4 expression was also examined by immunohistochemistry.. Our data revealed that the ratio of trypsinogen to that of the protective pancreas-specific trypsin inhibitor was elevated upon cigarette smoke exposure. The expression of carbonic anhydrase and CFTR remained unaltered when inflammatory signs were not detected in histological examinations. On the other hand, when pancreatic inflammation was present, the levels of CFTR and carbonic anhydrase were increased, indicating ductal and/or centroacinar cell involvement. No changes in the expression of Muc1 and Muc4 mucins were observed.. Our data show that cigarette smoke exposure leads to an increased vulnerability to pancreatic self-digestion. Moreover, the concomitant involvement of pancreatic ducts occurs only when focal pancreatic inflammation is present.

Topics: Animals; Carbonic Anhydrases; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Gene Expression Regulation; Mucins; Pancreas, Exocrine; Pancreatic Ducts; Pancreatitis; Rats; Rats, Sprague-Dawley; Receptor, Cholecystokinin A; RNA, Messenger; Tobacco Smoke Pollution; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Amino Acid Substitution; Humans; Mutation, Missense; Pancreatitis; Trypsin; Trypsinogen

The identification of a specific mutation in the human cationic trypsinogen gene in large kindreds with hereditary pancreatitis was the key to understand the genetic background of chronic pancreatitis. Rapidly, other variants within the same gene were identified-even in small families with a minority of patients. Later, mutations of the most important intrapancreatic trypsin inhibitor SPINK1 were found with high prevalence in patients with idiopathic, tropical and alcoholic chronic pancreatitis. We summarize interesting genetic and biochemical findings, point to clinical features and review recommendations for genetic analysis, follow-up and cancer prevention.

Topics: Carrier Proteins; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Genetic Predisposition to Disease; Genotype; Humans; Pancreas; Pancreatitis; Phenotype; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Hereditary pancreatitis is a rare autosomal dominant inherited disease with 80% penetration rate. The disease is characterized by recurrent episodes of pancreatitis often beginning in childhood, positive family history with at least two other affected members and no known precipitating factors. Most forms of hereditary pancreatitis are caused by one of two commoner mutations, R122H in exon 3 and N29I in exon 2 of the cationic trypsinogen (CT) (PRSS1) gene, located on chromosome 7. These genetic defects are speculated to cause excessive trypsin activity or to prevent inactivation of prematurely activated trypsin, resulting in pancreatitis. We performed mutation analysis of a Korean family with two members having clinically suspicious hereditary pancreatitis. We analyzed the CT gene in DNA samples extracted from peripheral blood of five family members. First of all, polymerase chain reaction and restriction enzyme digestion were performed in exon 3 of the CT gene. And then DNA products were purified and sequenced. We found out that three members of the family, the mother and two daughters, had a R122H mutation of the CT gene. We report the first family of hereditary pancreatitis associated with the CT gene mutation, an arginine to histidine amino acid substitution at residue 122, in Korea.

Topics: Amino Acid Substitution; Child; DNA Mutational Analysis; Female; Gastrointestinal Hemorrhage; Humans; Mutation; Pancreatic Pseudocyst; Pancreatitis; Trypsin; Trypsinogen

Hereditary pancreatitis (HP) is an autosomal dominant inherited disease characterized by recurrent episodes of pancreatitis often beginning in childhood, a family history of at least 2 other affected members, and the absence of known etiologic factors. The discovery of mutations in cationic trypsinogen gene (PRSS1) in HP not only provided insights into the molecular mechanisms of pancreatitis, but also opened a new era in the field of chronic pancreatitis. The detection of mutations in serine protease inhibitor, Kazal type 1 (SPINK1) and CFTR in patients with hereditary or idiopathic chronic pancreatitis has placed the emphasis on the importance of genetic mutations in pancreatitis. Because the estimated cumulative risk of pancreatic cancer development in hereditary pancreatitis is nearly 40%, screening tests are important in selected cases. There are no specific medical therapies recommended in patients with HP. Registration of patients with Nationwise Registries is essential if management strategies are to be improved and genetic research to be continued.

Topics: Carrier Proteins; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

To assess the effect of non-selective ET(A/B) (LU 302872) and selective ET(A) (LU 302146) antagonist on pancreatic histology and ultrastructure of acinar cells in connection with trypsinogen activation in early caerulein-induced AP.. Male Wistar rats with caerulein-induced AP, lasting 4 h, were treated i.p. with 10 and 20 mg/kg b.w. of each antagonist. Edema, inflammatory infiltration, necrosis and vacuolization of acinar cells in the pancreas were scored at 0-3 scale. Free active trypsin (FAT), total potential trypsin (TPT) after activation with enterokinase, and index of trypsinogen activation (%FAT/TPT) were assayed in pancreatic homogenates.. In untreated AP, the edema, inflammatory infiltration, necrosis and vacuolization increased as compared to control healthy rats (P<0.01). None of the treatment exerted any meaningful effect on the edema and inflammatory infiltration. The selective antagonist increased slightly the necrosis score to 0.82+/-0.06 at higher dose (P<0.05) vs 0.58+/-0.06 in untreated AP. The non-selective antagonist increased slightly the vacuolization score to 2.41+/-0.07 at higher dose (P<0.01) vs 1.88+/-0.08 in untreated AP. The decrease in the number of zymogen granules, disorganization of endoplasmic reticulum, autophagosomes and cytoplasmic vacuoles were more prominent in treated AP than in untreated AP groups. %FAT/TPT in untreated AP increased about four times (18.4+/-3.8 vs 4.8+/-1.3 in control group without AP, P<0.001). Treatment of AP with both antagonists did not affect significantly augmented trypsinogen activation.. The treatment with endothelin-1 receptors (non-selective ET(A/B) and selective ET(A)) antagonists has essential effect neither on the edema and inflammatory infiltration nor on trypsinogen activation observed in the early course of caerulein-induced AP. Nevertheless a slight increase of the necrosis and vacuolization score and some of the ultrastructural data could suggest the possibility of their undesired effects in caerulein-induced AP at investigated doses.

Topics: Acute Disease; Animals; Benzhydryl Compounds; Ceruletide; Endothelin A Receptor Antagonists; Male; Microscopy, Electron; Pancreas; Pancreatitis; Propionates; Pyrimidines; Rats; Rats, Wistar; Trypsinogen

Gene conversion--the substitution of genetic material from another gene--is recognized as the underlying cause of a growing number of genetic diseases. While in most cases conversion takes place between a normal gene and its pseudogene, here we report an occurrence of disease-associated gene conversion between two functional genes. Chronic pancreatitis in childhood is frequently associated with mutations of the cationic trypsinogen gene (serine protease 1; PRSS1). We have analyzed PRSS1 in 1106 patients with chronic pancreatitis, and identified a novel conversion event affecting exon 2 and the subsequent intron. The recombination replaced at least 289 nucleotides with the paralogous sequence from the anionic trypsinogen gene (serine protease 2; PRSS2), and resulted in the PRSS1 mutations c.86A > T and c.161A > G, causing the amino acid substitutions N29I and N54S, respectively. Analysis of the recombinant N29I-N54S double mutant cationic trypsinogen revealed increased autocatalytic activation, which was solely due to the N29I mutation. In conclusion, we have demonstrated that gene conversion between two functional paralogous trypsinogen genes can occur and cause genetically determined chronic pancreatitis.

Topics: Adolescent; Base Sequence; Child; Female; Gene Conversion; Humans; Models, Genetic; Molecular Sequence Data; Pancreatitis; Sequence Homology, Nucleic Acid; Trypsin; Trypsinogen

To investigate mutation of serine protease 1-cationic trypsinogen (CT, PRSS1) gene in members of a Thai family with hereditary pancreatitis and pancreatic cancer.. Polymerase chain reaction and direct sequencing were performed to analyze the PRSS1 gene in two members of the family affected by pancreatitis. Allele specific amplification (ASA) method was then developed to detect the mutation of the PRSS1 gene in all available members of the family and normal control subjects.. A cytosine (C) to thymine (T) mutation at position 2441 (g.2441C>T) of the PRSS1 gene, which results in a substitution of arginine by cysteine at position 116 (R116C) of CT, was identified by direct sequencing in both clinically affected members of the family but was not found in the unaffected member. This mutation, which might be arising from deamination of methylated cytosine in CpG dinucleotide of codon 116 (CGT>TGT), was also detected by the ASA method in the two affected members and a proband's brother but was not observed in unaffected members and 54 normal control subjects.. Autosomal dominant pancreatitis with increased cancer risk in the studied Thai family is most likely due to missense (R116C) mutation in the PRSS1 gene.

Topics: Exons; Family Health; Female; Genetic Predisposition to Disease; Humans; Middle Aged; Mutation, Missense; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Thailand; Trypsin; Trypsinogen

Topics: Calcinosis; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

Trypsinogen activation is thought to play a crucial role in the pathogenesis of acute pancreatitis (AP). Our aim was to characterize the very early sequential changes of trypsinogen-1, trypsinogen-2, the trypsin-2-alpha1-antitrypsin complex (T2-AAT), and pancreatic secretory trypsin inhibitor (PSTI) in serum from patients with pancreatitis induced by endoscopic retrograde cholangiopancreatography (ERCP), a model for studying the early phase of the disease in humans.. The study population consisted of 659 consecutive patients with 897 ERCP procedures. Blood samples were obtained before and at different time points after the procedure. The serum concentrations of trypsinogen-1 and trypsinogen-2, PSTI and T2-AAT were determined by time-resolved immunofluorometric assays.. ERCP-induced pancreatitis developed after 50 of the 897 ERCP procedures (5.6%). Sixty-one randomly selected ERCP patients without post-ERCP pancreatitis served as controls. Trypsinogen-1 and trypsinogen-2 showed an equally steep increase during the two first hours after ERCP in patients developing AP, but trypsinogen-1 decreased more rapidly than trypsinogen-2, which remained elevated during the 5-day study period. Serum PSTI also increased rapidly whereas T2-AAT increased more slowly peaking at 24 h. In patients developing post-ERCP pancreatitis the median concentration of trypsinogen-1 was markedly higher than in the controls already before the ERCP procedure. In the control group the concentrations of trypsinogen-1, trypsinogen-2, PSTI and T2-AAT did not change significantly.. The rapid increase of trypsinogen-1 and trypsinogen-2 and PSTI in the early phase of AP suggests that release of pancreatic enzymes is the initial event while the delayed increase of T2-AAT may reflect that the capacity of the intrapancreatic PSTI-based inhibitory mechanism has been exhausted.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin; Biomarkers; Carrier Proteins; Cholangiopancreatography, Endoscopic Retrograde; Early Diagnosis; Female; Humans; Intercellular Signaling Peptides and Proteins; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Severity of Illness Index; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Chronic pancreatitis: Only recently mutations in several genes were found in patients with chronic pancreatitis. In those with a familial chronic pancreatitis mutations of the cationic trypsinogen were identified and the variants N29I and R122H lead to an autosomal dominant disease. In this group of patients the mutation N34S of the trypsin inhibitor SPINK1 was detected. In so-called idiopathic pancreatitis both variants of the SPINK1 and of the CFTR (cystic fibrosis transmembrane conductance regular) were identified. Alterations in both genes were also found in patients with alcoholic chronic pancreatitis. The strongest risk factor for chronic pancreatitis were trypsinogen mutations N29I and R122H mutations. However, both SPINK1 and CFTR increased the risk for chronic pancreatitis to a higher level than alcohol consumption. A genetic investigation should be performed in familial disease and younger age, but also in patients without family history and higher age a mutation could be found. Pancreas cancer: In 10% of the patients with pancreas cancer other members of the family were affected from the disease. Some of them belong to well characterized familial syndroms like HNPCC or Peutz-Jeghers-syndrom. In a minority of the others a genetic factor may be found, too. In sporadic disease the development of the tumor is characterized by continued acquirement of genetic alterations described by the PanIN model (pancreatic intraepithelial neoplesia). This means that the evolution of the neoplasia progresses from normal tissue via epithelial hyperplasy (PanIN 1A), papillary hyperplasy without (PanIN 1B) and with dysplasy (PanIN 2) and carcinoma in situ (PanIN 3) to invasive pancreas cancer. The progression is associated with genetic alterations of the cells (mutations of ki-ras, p16, p53 etc.). This results in deterioration of control of the cell cycle and the apoptosis and explains the malignancy of the disease. These findings may be used in the future to develop newer therapeutic principles in order to improve the dismal prognosis of this disease.

Topics: Carrier Proteins; Cell Transformation, Neoplastic; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Disease Susceptibility; DNA Mutational Analysis; Genes, Dominant; Genotype; Humans; Pancreatic Neoplasms; Pancreatitis; Pancreatitis, Alcoholic; Risk Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

To assess the balance between trypsin and protease inhibitors simultaneously in the systemic circulation and in the thoracic lymph and peritoneal exudate.. Twenty patients with early severe acute pancreatitis were studied. Enzymatically active and immunoreactive trypsin in conjunction with its major inhibitors were measured in the 3 compartments at the onset of end-organ failure(s). The molecular forms of trypsin were determined in the lymph and ascites by gel filtration chromatography to separate trypsinogen and free-and inhibitor-bound trypsin.. Both enzymatically active trypsin and immunoreactive trypsin levels were highest in ascites and lymph compared with the systemic circulation. Intracompartmental alpha1- protease inhibitor gradient moved in the opposite direction, whereas alpha2 macroglobulin concentration was highest in ascites and lowest in the lymph. Although most of the enzymatically and immunoreactive material in ascites and lymph consisted of trypsin complexed with alpha2 macroglobulin and trypsinogen, respectively, free active trypsin was detected in more than 80% of the samples.. In patients with early severe acute pancreatitis, there is a significant trypsinogen activation resulting in protease-antiprotease imbalance and thereby free enzymatically active trypsin in the 2 body fluid compartments in close vicinity to the inflammatory process. This may be involved in the pathophysiology of local and distant tissue damage.

Topics: Acute Disease; Adult; Aged; alpha-Macroglobulins; Ascites; Body Fluid Compartments; Enzyme Activation; Female; Humans; Lymph; Male; Middle Aged; Pancreatitis; Peptide Hydrolases; Protease Inhibitors; Severity of Illness Index; Trypsin; Trypsinogen

Acute pancreatitis is characterized by the pathologic activation of zymogens within pancreatic acinar cells. The process requires a rise in cytosolic Ca(2+) from undefined intracellular stores. We hypothesized that zymogen activation is mediated by ryanodine receptor (RYR)-regulated Ca(2+) release, because early zymogen activation takes place in a supranuclear compartment that overlaps in distribution with the RYR. Ca(2+) signals in the basolateral, but not apical, region of acinar cells observed during supraphysiologic agonist stimulation were dependent on RYR Ca(2+) release. Inhibition of RYR or depletion of RYR-sensitive Ca(2+) pools each reduced pathologic zymogen activation in isolated acinar cells, but neither treatment affected amylase secretion. Inhibition of RYR also inhibited zymogen activation in vivo. We propose that Ca(2+) release from the RYR mediates zymogen activation but not enzyme secretion. The findings imply a role for the RYR in acute pancreatitis.

Topics: Animals; Calcium; Ceruletide; Dantrolene; Enzyme Precursors; Male; Microscopy, Confocal; Models, Biological; Pancreas, Exocrine; Pancreatitis; Rats; Rats, Sprague-Dawley; Ryanodine Receptor Calcium Release Channel; Secretory Vesicles; Trypsinogen

Topics: Adult; Chronic Disease; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Mutation; Pancreatitis; Pedigree; Trypsinogen

Heat shock proteins (Hsp's) protect cellular proteins in response to injury, and the role of Hsp70 in experimental pancreatitis was recently described. To find out the possible role of Hsp70 in pancreatitis, we used Hsp70 knock-out mice (Hsp70.1-/-) and wild-type mice (Hsp70.1+/+).. We studied enzymes activities, Hsp70 protein levels, and histologies in cerulein-induced pancreatitis of Hsp70.1-/- and Hsp70.1+/+ mice.. In the basal state, Hsp70 protein levels were higher in Hsp70.1+/+ than in Hsp70.1-/- mice, and trypsin activity was higher in Hsp70.1-/- than in Hsp70.1+/+ mice. The zymogen/lysosome ratio of cathepsin B activity before cerulein injection was higher in Hsp70.1-/- than in Hsp70.1+/+ mice. The expression level of Hsp70 in the pancreas increased in both of Hsp70.1-/- and Hsp70.1+/+ mice after hyperthermia because of the Hsp70.3 gene left intact in Hsp70.1-/- mice. After cerulein hyperstimulation, trypsin activity increased 2-fold in Hsp70.1+/+ mice, but cerulein did not further increase basally elevated trypsin activity in Hsp70.1-/- mice. Hyperthermia pretreatment not only blocked cerulein-induced trypsinogen activation, pancreatic edema, and vacuolization in Hsp70.1+/+ mice, but also decreased basally elevated trypsin activity in Hsp70.1-/- mice.. Hsp70 can be responsible for inhibition of cerulein-induced pancreatitis and prevention of spontaneous trypsinogen activation in mice by inhibiting the colocalization of zymogen and lysosomal enzymes.

Topics: Animals; Cathepsin B; Enzyme Activation; Fever; HSP70 Heat-Shock Proteins; Mice; Mice, Knockout; Pancreas; Pancreatitis; Trypsinogen

To assess effects of NF-kappaB activation inhibitor (pyrrolidine dithiocarbamate--PDTC) alone or with endothelins (ET-1, ET-2, ET-3) in early course of cerulein-induced acute pancreatitis (AP) in rats.. After 4 h of AP in Wistar rats, treated with PDTC 10 or 40 mg/kg or with PDTC 10 mg/kg and ET-1, ET-2 or ET-3, 0.5 or 1.0 nmol/kg twice i.p. in 1 h interval, free active trypsin (FAT), total potential trypsin (TPT) and lipase in 12000 x g supernatants of pancreatic homogenates, plasma alpha-amylase and histological changes were assayed. %FAT/TPT was an index of trypsinogen activation.. %FAT/TPT significantly increased to 12.42 +/- 2.14%, lipase to 5.51 +/- 0.84 U/mg protein and alpha-amylase to 28.5 +/- 5.61 U/mL in AP vs 1.96 +/- 0.31%, 1.29 +/- 0.11 U/mg and 5.80 +/- 1.38 U/ml in healthy control. Higher dose PDTC attenuated trypsinogen activation to 3.01 +/- 0.53% and alpha-amylase to 15.3 +/- 1.38. PDTC and ET-1 attenuated %FAT/TPT to 2.55 +/- 0.18% with lower and 2.34 +/- 0.44% with higher dose. ET-3 was less effective than ET-1: 6.76 +/- 0.46% with lower dose. Lower doses of ET-1 and ET-2 with PDTC, diminished lipase activity to 2.60 +/- 0.36 and 2.94 +/- 0.33.. Cumulative attenuation of trypsinogen activation after lower dose of PDTC and ET-1 approximated the effect of higher dose of PDTC. Additional effect of ET-3 was weaker than ET-1, and ET-2 was ineffective in this respect. The combination of this NF-kappaB activation inhibitor and ET-1 could be beneficial in early course of edematous AP by attenuating of trypsinogen activation. However, it should be treated with caution because of some unfavorable effects on histological scores of pancreatic injury.

Topics: Acute Disease; alpha-Amylases; Animals; Antioxidants; Ceruletide; Endothelin-1; Endothelin-2; Endothelin-3; Enzyme Activation; Lipase; Male; NF-kappa B; Pancreatitis; Pyrrolidines; Rats; Rats, Wistar; Thiocarbamates; Trypsinogen

To assess the usefulness of urinary trypsinogen-2 test strip, urinary trypsinogen activation peptide (TAP), and serum and urine concentrations of the activation peptide of carboxypeptidase B (CAPAP) in the diagnosis of acute pancreatitis.. Patients with acute abdominal pain and hospitalized within 24 h after the onset of symptoms were prospectively studied. Urinary trypsinogen-2 was considered positive when a clear blue line was observed (detection limit 50 microg/L). Urinary TAP was measured using a quantitative solid-phase ELISA, and serum and urinary CAPAP by a radioimmunoassay method.. Acute abdominal pain was due to acute pancreatitis in 50 patients and turned out to be extrapancreatic in origin in 22 patients. Patients with acute pancreatitis showed significantly higher median levels of serum and urinary CAPAP levels, as well as amylase and lipase than extrapancreatic controls. Median TAP levels were similar in both groups. The urinary trypsinogen-2 test strip was positive in 68% of patients with acute pancreatitis and 13.6% in extrapancreatic controls (P<0.01). Urinary CAPAP was the most reliable test for the diagnosis of acute pancreatitis (sensitivity 66.7%, specificity 95.5%, positive and negative predictive values 96.6% and 56.7%, respectively), with a 14.6 positive likelihood ratio for a cut-off value of 2.32 nmol/L.. In patients with acute abdominal pain, hospitalized within 24 h of symptom onset, CAPAP in serum and urine was a reliable diagnostic marker of acute pancreatitis. Urinary trypsinogen-2 test strip showed a clinical value similar to amylase and lipase. Urinary TAP was not a useful screening test for the diagnosis of acute pancreatitis.

Topics: Adult; Aged; Biomarkers; Case-Control Studies; Female; Humans; Male; Middle Aged; Oligopeptides; Pancreatitis; Peptides; Predictive Value of Tests; Trypsin; Trypsinogen

The human pancreas secretes two major trypsinogen isoforms, cationic and anionic trypsinogen. To date, 19 genetic variants have been identified in the cationic trypsinogen gene (PRSS1) of patients with hereditary, familial, or sporadic chronic pancreatitis. A common feature of cationic trypsinogen mutants studied so far is an increased propensity for autocatalytic activation (autoactivation). This is thought to lead to premature intrapancreatic digestive protease activation. In contrast, no pancreatitis-associated mutations have been found in the anionic trypsinogen gene (PRSS2), suggesting that this isoform might play a relatively unimportant role in pancreatitis. To challenge this notion, here we describe the unique properties of the E79K cationic trypsinogen mutation (c.235G>A), which was identified in three European families affected by sporadic or familial pancreatitis cases. In vitro analysis of recombinant wild-type and mutant enzymes revealed that catalytic activity of E79K trypsin was normal, and its inhibition by pancreatic secretory trypsin inhibitor was unaffected. Although the E79K mutation introduces a potential new tryptic cleavage site, autocatalytic degradation (autolysis) of E79K-trypsin was also unchanged. Furthermore, in contrast to previously characterized disease-causing mutations, E79K markedly inhibited autoactivation of cationic trypsinogen. Remarkably, however, E79K trypsin activated anionic trypsinogen two-fold better than wild-type cationic trypsin did, while the common pancreatitis-associated mutants R122H or N29I had no such effect. The observations not only suggest a novel mechanism of action for pancreatitis-associated trypsinogen mutations, but also highlight the importance of interactions between the two major trypsinogen isoforms in the development of genetically determined chronic pancreatitis.

Topics: Adult; Aged; Carrier Proteins; Catalysis; Cathepsins; Chronic Disease; DNA Mutational Analysis; Enzyme Activation; Female; Humans; Intercellular Signaling Peptides and Proteins; Isoenzymes; Kinetics; Male; Middle Aged; Mutation; Pancreatitis; Pedigree; RNA, Messenger; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Mutation of Cationic trypsinogen gene is clearly associated with hereditary pancreatitis and plays an important role in the pathogenesis of pancreatitis. According to literature, this mutation is occasionally occurred in patients with pancreatitis in Western countries and Japan. The aim of this study was to find out whether the mutation was observed in Korean patients with chronic idiopathic pancreatitis.. Peripheral blood samples of 11 patients with chronic idiopathic pancreatitis were collected consecutively, and DNA was extracted from the samples. Polymerase chain reaction was performed in exon 2 and 3 of cationic trypsinogen gene. Then, DNA products were purified and sequenced.. The mutation was not found in exon 2 and 3 of cationic trypsinogen gene in these patients.. There was no cationic trypsinogen mutation in Korean patients with chronic idiopathic pancreatitis. Further large sampled cohort study is needed.

Topics: Adolescent; Adult; Chronic Disease; Female; Humans; Male; Middle Aged; Mutation; Pancreatitis; Polymerase Chain Reaction; Trypsin; Trypsinogen

Hereditary pancreatitis is an autosomal dominant disease that is mostly caused by cationic trypsinogen (PRSS1) gene mutations. The aim was to determine phenotype-genotype correlations of families in Europe.. Analysis of data obtained by the European Registry of Hereditary Pancreatitis and Pancreatic Cancer was undertaken using multilevel proportional hazards modelling.. There were 112 families in 14 countries (418 affected individuals): 58 (52%) families carried the R122H, 24 (21%) the N29I, and 5 (4%) the A16V mutation, 2 had rare mutations, and 21 (19%) had no PRSS1 mutation. The median (95% confidence interval [CI]) time to first symptoms for R122H was 10 (8, 12) years of age, 14 (11, 18) years for N29I, and 14.5 (10, 21) years for mutation negative patients (P = 0.032). The cumulative risk (95% CI) at 50 years of age for exocrine failure was 37.2% (28.5%, 45.8%), 47.6% (37.1%, 58.1%) for endocrine failure, and 17.5% (12.2%, 22.7%) for pancreatic resection for pain. Time to resection was significantly reduced for females (P < 0.001) and those with the N29I mutation (P = 0.014). The cumulative risk (95% CI) of pancreatic cancer was 44.0% (8.0%, 80.0%) at 70 years from symptom onset with a standardized incidence ratio of 67% (50%, 82%).. Symptoms in hereditary pancreatitis start in younger patients and endpoints take longer to be reached compared with other forms of chronic pancreatitis but the cumulative levels of exocrine and endocrine failure are much higher. There is an increasingly high risk of pancreatic cancer after the age of 50 years unrelated to the genotype.

Topics: Adult; Age Distribution; Age of Onset; Confidence Intervals; Europe; Female; Genetic Diseases, Inborn; Genetic Predisposition to Disease; Heterozygote; Humans; Incidence; Male; Middle Aged; Multivariate Analysis; Pancreatitis; Pedigree; Point Mutation; Probability; Prognosis; Registries; Reproducibility of Results; Risk Assessment; Severity of Illness Index; Sex Distribution; Survival Rate; Trypsin; Trypsinogen

Hereditary pancreatitis (HP) is the strongest known risk factor for pancreatic cancer. The aim of the present study is to establish diagnostic criteria for HP to predict and identify high-risk groups for pancreatic cancer.. We collected clinical data for 210 patients with recurrent acute or chronic pancreatitis, and examined mutations of the cationic trypsinogen (CT) gene in 57 patients with a family history of pancreatitis or with early-onset idiopathic recurrent acute or chronic pancreatitis (40 years of age or younger). DNA was extracted from peripheral blood leukocytes, and exons 2 and 3 of the CT gene were individually amplified by polymerase chain reaction (PCR) and sequenced.. Of these 57 patients in whom mutations of the CT gene were examined, the R122H (20 patients) and N29I (5 patients) mutations in the CT gene were observed in 25 patients (43.9%). From the analysis of clinical records and the CT gene of these patients, we proposed the following adaptations to the diagnostic criteria for HP: (1) at least one of the affected members in a family has no known etiological factors, (2) we deleted the definition of "different generation", but included the upper limit of the age of onset of pancreatitis in the case of siblings (at least 1 of the patients in a family <40 years of age). According to these criteria, all patients with the CT gene mutations in the present study could be classified as having HP, with the exception of 2 sporadic cases with the R122H and N29I mutations, respectively. Based on these findings, we revised the criteria for the diagnosis of HP; (1) recurrent acute or chronic pancreatitis with R122H or N29I mutation of the CT gene, or (2) recurrent acute or chronic pancreatitis with a family history of 2 or more affected patients, irrespective of generation, with at least 1 of the patients having no known etiological factors, and in case of siblings only, the onset of the disease in at least 1 of the patients is under age 40 years.. The revised criteria in the present study are appropriate and of clinical usefulness to diagnose patients with HP even in cases without the genetic testing. However, if and when more genes are detected, it will be important to reexamine the mutation-negative patients now classified as HP based on our proposed criteria.

Topics: Adolescent; Adult; Age of Onset; Aged; Child; DNA Mutational Analysis; Female; Humans; Japan; Male; Middle Aged; Pancreatitis; Trypsin; Trypsinogen

Mutations in the cationic trypsinogen (protease, serine, 1 (trypsin 1); PRSS1) gene are causally associated with recurrent acute and chronic pancreatitis. We investigated whether mutations in the PRSS1 gene are associated with hereditary and non-hereditary pancreatitis. As a modifier role has been proposed for trypsin inhibitor (serine protease inhibitor, Kazal type I; SPINK1) mutations, the role of SPINK1 mutations in these patients was also analysed.. The coding regions of PRSS1 and SPINK1 genes were sequenced in 290 controls and 198 patients, of whom 120 were diagnosed as idiopathic (ICP), 41 as alcoholic (ACP), and 37 as hereditary pancreatitis (HP). Twenty four unaffected relatives of HP probands were also analysed and genotype-phenotype correlations and statistical analyses were performed.. No mutations in the PRSS1 gene were detected in any of the patients, including HP patients, while the N34S mutation was observed in the SPINK1 gene in the majority of HP patients (73%). Similarly, 26.8% of ACP (11 of 41) and 32.5% (39 of 120) of ICP patients also had SPINK1 mutations. The N34S mutation was observed in both homozygous and heterozygous conditions. In comparison, only 2.76% of the control population had the N34S allele (p<0.001). The P55S mutation was observed in one ICP and one ACP patient, and in three normal individuals. Genotype-phenotype correlations did not suggest any significant difference in the age of onset, severity of disease, or pancreatic endocrine insufficiency in patients with or without mutated SPINK1 and irrespective of the allelic status of N34S SPINK1.. Irrespective of the aetiology, mutations in the PRSS1 gene are not associated with chronic pancreatitis, including HP. In contrast, the N34S mutation in the SPINK1 gene shows a significant correlation in these patients. A comparable phenotype in terms of age of onset, diabetes mellitus, and other phenotypic features in patients with or without SPINK1 mutations and N34S homozygotes and heterozygotes suggests that there may still be involvement of other genetic or environmental factors.

Topics: Adult; Chronic Disease; DNA Mutational Analysis; Female; Genetic Predisposition to Disease; Genotype; Humans; Male; Middle Aged; Mutation; Pancreatitis; Pedigree; Phenotype; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

The effects of stable prostacyclin analogue iloprost on the trypsinogen activation, labilization of lysosomal membranes, lipolytic enzymes activities, histopathological and ultrastructural changes in the pancreas of rats with severe, taurocholate acute pancreatitis (AP), preceded for 6 h by acute ethanol intake have been investigated. Iloprost (1 microg/kg b.w., i.p.) was applied every 6 hours after inducing of taurocholate AP. The antecedent intragastric 40% ethanol intake (5 g/kg b.w.) increased an index of trypsinogen activation in AP lasting 18 h. Treatment with iloprost prevented this increase in the rats with AP given earlier alcohol, and limited the labilization of lysosomal membranes in nonalcoholized rats with AP. Phospholipase A2 and lipase activities were reduced by iloprost only in the rats not given ethanol. The additional damaging effect of acute ethanol abuse prior to AP could be dependent on augmented activation of trypsinogen. The protective effect of iloprost in AP seems to be dependent on the attenuation of trypsinogen activation, decrease of total potential trypsin and the decrease of lysosomal membranes labilization. Its protective effect could be limited in taurocholate acute pancreatitis preceded by acute ethanol intake as evidenced by the differences in the cathepsin B, phospholipase A2 and lipase activities and by histopathological and ultrastructural examination.

Topics: Acute Disease; Animals; Disease Models, Animal; Ethanol; Iloprost; Intracellular Membranes; Lipase; Lysosomes; Male; Pancreas; Pancreatitis; Phospholipases A; Phospholipases A2; Rats; Rats, Wistar; Taurocholic Acid; Trypsinogen

Severe impairment of exocrine pancreatic secretion has recently been demonstrated in a clinical study in sepsis and septic shock patients. The purpose of this study was to further evaluate involvement of the pancreas in the acute phase reaction in sepsis. Using a normotensive rat model of Pseudomonas pneumonia-induced sepsis, we assessed the expression of PAP-I, amylase and trypsinogen mRNA, PAPI protein levels, and cytokine expression in the pancreas by Northern and Western blot analysis and RT-M PCR, respectively. Presence of several well-established features of pancreatitis in sepsis-induced animals were examined by biochemical and histopathological methods as well as by a determination of both water and myeloperoxidase content. Sepsis resulted in an up-regulation of PAP-I gene expression and increase in its protein level in pancreas while the mRNA levels of amylase and trypsinogen were down-regulated. Differences in the pancreatic cytokine expression, serum amylase and serum lipase levels, the occurrence of pancreatic edema as well as the severity of inflammatory infiltration and necrosis were not significantly different between sham and pneumonia groups. Acinar cells showed increased vacuolization in pneumonia animals 24 hours after the treatment. These findings demonstrate that the pancreas is actively involved in the acute phase reaction in sepsis of remote origin. This involvement occurs without concomitant biochemical and histopathologic alterations observed in pancreatitis. Taken all together, these features are indicative of a sepsis-specific dysfunction of the pancreas.

Topics: Acute-Phase Reaction; Amylases; Animals; Antigens, Neoplasm; Biomarkers, Tumor; Cytokines; Gene Expression Regulation; Lectins, C-Type; Leukocyte Count; Lipase; Male; Necrosis; Pancreas; Pancreatitis; Pancreatitis-Associated Proteins; Peroxidase; Pneumonia, Bacterial; Pseudomonas Infections; Rats; Rats, Sprague-Dawley; RNA, Messenger; Sepsis; Time Factors; Trypsinogen; Vacuoles

Serum and urine concentrations of the activation peptide of carboxypeptidase B (CAPAP) and urinary trypsinogen activation peptide (TAP) as prognostic markers in acute pancreatitis were compared.. Fifty-two patients with acute pancreatitis hospitalized within 24 hours after symptom onset were prospectively studied. Blood and urine samples were obtained during the first 3 days of the hospital stay.. Pancreatitis was severe in 17 patients and mild in 35 (Atlanta criteria). Median serum CAPAP levels on days 1 and 2 and of urine CAPAP and TAP on days 1, 2, and 3 were significantly higher in severe pancreatitis than in mild disease. On the first day of admission, TAP was the most accurate predictor of severity (sensitivity, 92.3%; specificity, 80%; positive and negative predictive values, 63.2% and 96.6%, respectively), with a 4.61 positive likelihood ratio for a cutoff value of 18.10 nmol/L, whereas within 24 hours after symptom onset, urinary CAPAP was superior (sensitivity, 88.9%; specificity, 81.3%; positive and negative predictive values 72.7% and 92.9%, respectively), with a 4.72 positive likelihood ratio for a cutoff value of 15.45 nmol/L.. Serum and urine CAPAP levels and urinary TAP are accurate in the early assessment of severity in acute pancreatitis. Urine CAPAP levels was the most accurate marker 24 hours after onset of symptoms.

Topics: Abdominal Pain; Acute Disease; Adult; Aged; Biomarkers; Carboxypeptidase B; Enzyme Activation; Female; Humans; Male; Middle Aged; Oligopeptides; Pancreatitis; Peptides; Predictive Value of Tests; Prognosis; Prospective Studies; ROC Curve; Severity of Illness Index; Trypsinogen

The clinical usefulness of urinary trypsinogen-2 dipstick test is still in controversy. We evaluated the usefulness of urinary trypsinogen-2 dipstick test in patients with acute pancreatitis.. Urinary trypsinogen-2 dipstick test was prospectively performed in 50 patients with acute pancreatitis, 50 patients with non-pancreatic abdominal pain, and 50 healthy controls.. On admission, urinary trypsinogen-2 dipstick test was positive in 36 of 50 patients with acute pancreatitis (sensitivity, 72%) and in 4 of 50 patients with non-pancreatic abdominal pain (specificity, 92%). On the other hand, it was all negative in controls. The sensitivity and specificity of serum lipase were 78% and 94%, respectively. At 24 hours after admission, the positive rate of urinary trypsinogen-2 dipstick test rose from 72% to 94% (p=0.02). The results of urinary trypsinogen-2 dipstick test was positive in 14 of 15 patients with severe pancreatitis and 22 of 35 patients with mild pancreatitis according to the criteria by Atlanta International Symposium, 1992.. Urinary trypsinogen-2 dipstick test is comparable to serum lipase in diagnosing acute pancreatitis. Delayed measurement and severe pancreatitis are more likely to yield positive results with urinary trypsinogen-2 dipstick test. Thus, we suggest that the cut-off value of urinary trypsinogen-2 dipstick test should be lowered to increase its sensitivity.

Topics: Acute Disease; Adult; Aged; Biomarkers; Female; Humans; Lipase; Male; Middle Aged; Pancreatitis; Reagent Strips; Sensitivity and Specificity; Trypsinogen

Hereditary pancreatitis is an autosomal dominant condition characterized by recurrent episodes of acute pancreatitis, usually starting in childhood. We present a family who was ascertained when an 11-year-old girl presented with an episode of acute pancreatitis. Her father and other family members had also had recurrent bouts of acute pancreatitis. Genetic testing revealed a pathogenic mutation in the cationic trypsinogen gene in the proband, her father and her paternal grandmother. As far as we are aware, this is the first Aboriginal kindred with mutation-proven hereditary pancreatitis. Hereditary pancreatitis is an important differential diagnosis to consider in a patient with recurrent episodes of acute pancreatitis with no obvious precipitating cause. This family is of Aboriginal descent and the implications of the family's background are also discussed when considering the aetiology of the condition. We emphasize the need to ascertain a full family history from patients with a history of repeated episodes of acute pancreatitis and also emphasize the need to avoid ethnic stereotypes when assessing patients.

Topics: Child; Exons; Family; Female; Genetic Diseases, Inborn; Humans; Mutation; Native Hawaiian or Other Pacific Islander; Pancreatitis; Trypsinogen

It has been found that mutations of cationic trypsinogen gene (PRSS1) and serine protease inhibitor, Kazal type 1 gene (SPINK1) increase the susceptibility of chronic pancreatitis (CP). Specifically, mutations in the PRSS1 gene are related to the occurrences of hereditary and idiopathic pancreatitis while SPINK1 mutations are known to act as a disease modifier and are associated with idiopathic CP. However, the association of SPINK1 mutations with alcoholic CP is still controversial. We investigated the prevalence of PRSS1 and SPINK1 mutations in idiopathic and alcoholic CP in Korea.. Seventy-one Korean patients with CP (alcoholic: 47, idiopathic: 22 and familial: 2) and 19 controls were included in this studies. Genomic DNA was extracted from peripheral blood of the patients. Mutations of SPINK1 (exon 3: N34S) and PRSS1 (exon 2: N29I, exon 3: R122H) genes were detected by PCR-RFLP methods. For the detection of SPINK1 mutation, restriction endonuclease PstI and BsrDI were used, while Sau3A and AflIII were used for the defection of PRSS1 mutation.. Only one patient (2.1%) with alcoholic CP was a heterozygote for SPINK1 (N34S) mutation. Mutation in the PRSS1 (N29I, R122H) gene was not found in any group of CP patients. Additionally, we could not find any mutations of SPINK1 or PRSS1 in the control group.. SPINK1 and PRSS1 mutations are not related to the development of CP in Korea.

Topics: Carrier Proteins; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Mutation; Pancreatitis; Pancreatitis, Alcoholic; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Specific mutations in the cationic trypsinogen gene ( PRSS1) are disease-causing in patients with hereditary pancreatitis, but the genetic background still remains mysterious in about 40% of patients with the disease. It has been suggested that oxidative stress contributes to pancreatic damage. The glutathione s-transferases (GSTs) represent major detoxification enzymes that protect cells from oxidative stress.. In the present study we tested whether mutations in the MGST1 and GSTM3 genes or common deletions in the GSTT1 and GSTM1 genes are associated with hereditary pancreatitis. We analyzed the entire coding region of MGST1 and GSTM3 in 30 patients that were tested negative for PRSS1 mutations, and we studied 55 controls. For GSTT1 and GSTM1, we investigated 75 hereditary pancreatitis patients who had been tested negative for PRSS1 mutations, 135 hereditary pancreatitis patients with a PRSS1 mutation, and 183 controls. Patients were further subclassified with regard to age of onset of disease as a marker of severity.. No mutation was found in the MGST1 gene. In the GSTM3 gene, we detected a homozygous 670G > A polymorphism (V224I) with similar frequencies in patients and controls. We found no difference in the frequencies of the GSTT1 and GSTM1 null genotypes between patients and controls, and we detected no differences in age of onset in patients with or without GSTT1 and GSTM1 deletions.. We conclude that genetic alterations in the MGST1, GSTM3, GSTT1, and GSTM1 genes do not play a dominant role in hereditary pancreatitis.

Topics: Adolescent; Adult; Age Factors; Aged; Child; Child, Preschool; Chronic Disease; DNA Mutational Analysis; Exons; Female; Gene Frequency; Genetic Predisposition to Disease; Glutathione Transferase; Homozygote; Humans; Infant; Male; Middle Aged; Open Reading Frames; Oxidative Stress; Pancreatitis; Polymerase Chain Reaction; Polymorphism, Genetic; Prognosis; Reference Values; Trypsin; Trypsinogen

The purpose of this study was to evaluate the cationic trypsinogen gene in miniature schnauzers for possible mutations. Genetic mutations have been linked with hereditary pancreatitis in humans. Four miniature schnauzers were selected on the basis of a clinical history of pancreatitis. One healthy miniature schnauzer and 1 healthy mixed breed canine were enrolled as controls. DNA was extracted from these canines using a commercial kit. Primers were designed to amplify the entire canine cationic trypsinogen cDNA sequence. A polymerase chain reaction (PCR) was performed and products were purified and sequenced. All sequences were then compared. The healthy control canine, a healthy miniature schnauzer, and the 4 miniature schnauzers with pancreatitis showed identical sequences of the cationic trypsinogen gene to the published sequence. We conclude that, in contrast to humans with hereditary pancreatitis, mutations of the cationic trypsinogen gene do not play a major role in the genesis of pancreatitis in the miniature schnauzer.

Topics: Amino Acid Sequence; Animals; DNA Mutational Analysis; DNA, Complementary; Dog Diseases; Dogs; Genetic Predisposition to Disease; Mutation; Pancreatitis; Polymerase Chain Reaction; Trypsinogen

To assess the effect of endothelins: ET-1, ET-2 and ET-3 on trypsinogen activation, lipase activity and histological changes in the pancreas in early (4 hrs) cerulein acute pancreatitis (AP) in rats.. In 45 Wistar rats with cerulein induced AP (2 x 40 microg/kg i.p. at 1 hour interval, the effect of endothelins at the dose 2 x 0.5 or 2 x 1.0 nmol/kg i.p. was assessed vs untreated AP; 6 healthy rats were control (C). Free active trypsin (FAT), total potential trypsin after activation with enterokinase (TPT), lipase in 12000 xg supernatants of pancreatic homogenates and the plasma alpha-amylase were assayed. The %FAT/TPT was an index of trypsinogen activation.. %FAT/TPT increased from 3.0 +/- 0.6 in C to 16.2 +/- 3.1 in AP (p < 0.01). ET-1 decreased this index to 4.8 +/- 1.1 after higher dose (p < 0.01); the effect of lower dose was insignificant. Attenuating effect of ET-2 was significant: 7.3 +/- 1.7 after higher dose (p < 0.05) and 6.1 +/- 0.9 after lower dose (p < 0.01). ET-3 diminished this index to 4.5 +/- 1.5 (p < 0.01) and to 6.3 +/- 2.2 (p < 0.05) respectively. Lipase activity in supernatant increased from 4.1 +/- 0.6 in C to 6.3 +/- 0.7 U/mg protein in untreated AP (p < 0.05) and plasma alpha-amylase from 7.0 +/- 0.6 in C to 25.9 +/- 4.3 U/ml in AP (p < 0.001), without essential changes in treated groups vs untreated AP. Higher doses of endothelins decreased inflammatory cell infiltration score in AP.. The exogenous endothelins, especially ET-2 and ET-3 and to lesser extent ET-1 exerted some protective effect in early, edematous acute pancreatitis by the attenuation of trypsinogen activation and inflammatory cell infiltration in the pancreas.

Topics: Acute Disease; alpha-Amylases; Animals; Ceruletide; Endothelin-1; Endothelin-2; Endothelin-3; Endothelins; Enzyme Activation; Lipase; Male; Pancreatitis; Rats; Rats, Wistar; Time Factors; Trypsinogen

We examined the effects of a weak base, chloroquine, on the trypsinogen processing in cerulein-induced pancreatitis.. Immunofluorescence studies were performed using newly generated affinity-purified antibodies to the trypsinogen activation peptide (TAP).. The present study showed that chloroquine pretreatment blocked intracellular TAP generation in cerulein-induced pancreatitis.. These results indicate that intracellular trypsinogen activation, which plays an important role in acute pancreatitis, requires a low-pH compartment, as well as serine protease activity.

Topics: Animals; Ceruletide; Chloroquine; Enzyme Inhibitors; Fluorescent Antibody Technique; Hydrogen-Ion Concentration; Male; Oligopeptides; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

Mutations of three major genes are associated with an increased risk of acute and chronic pancreatitis: the cationic trypsinogen (PRSS1) gene, the cystic fibrosis transmembrane conductance regulator (CFTR) gene, and the pancreatic secretory trypsin inhibitor (PSTI) or serine protease inhibitor, Kazal type 1 (SPINK1) gene. Some autosomal dominant forms of hereditary pancreatitis are associated with mutations of the PRSS1 gene, which can be readily identified by genetic testing. Mutations of the CFTR gene can lead either to cystic fibrosis or to idiopathic chronic pancreatitis, and to a variety of cystic fibrosis-associated disorders, including congenital bilateral absence of the vas deferens and sinusitis. These mutations, as with those of the SPINK1 (or PSTI) gene, are prevalent in North America; thus, the presence of such a mutation in an asymptomatic person does not confer a high risk of developing pancreatitis. Combinations of mutations of the PRSS1 and SPINK1 genes lead to more severe disease, as indicated by an earlier onset of symptoms, which suggests that SPINK1 is a disease modifier. The major fear expressed by potential candidates for genetic testing is that the results could lead to insurance discrimination. Studies of the positive predictive value of genetic tests are hampered by recruitment bias and lack of knowledge of family history of pancreatitis. Genetic testing is most useful for persons for whom family members have already been found to exhibit a particular pancreatitis-associated mutation. In the future, increased knowledge of the myriad genetic causes of pancreatitis, as well as advances in the diagnosis and treatment of early chronic pancreatitis, should enhance the utility of genetic testing.

Topics: Codon; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Testing; Humans; Mutation; Pancreatitis; Predictive Value of Tests; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Chronic Disease; Humans; Mutation; Pancreatitis; Trypsinogen

Topics: Chronic Disease; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

Topics: Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Pancreatitis; Trypsin; Trypsinogen

Topics: DNA; DNA Mutational Analysis; Family Health; Female; Humans; Male; Mutation; Mutation, Missense; Pancreatitis; Pedigree; Phenotype; Polymorphism, Genetic; Severity of Illness Index; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Biologic data related to pancreatic regeneration and acinar-cell homeostasis after ductal decompression would be useful in clinical settings to elucidate the time at which obstructions in human biliary acute pancreatitis (AP) should be removed. Our aim was to evaluate the outcome of AP after early removal of bile-pancreatic-duct obstruction (BPDO) and to ascertain whether cholecystokinin (CCK) blockade accelerates recovery from the disease. We conducted analysis of apoptosis and cell cycle, as well as measurements of enzyme and calcium load, in acinar cells using flow cytometry to ascertain the capability of the pancreas to regain its function after AP. Male Wistar rats were subjected to AP by means of BPDO for 6 hours and 24 hours. In other groups, the BPDO was opened 24 hours after induction; 3 days and 7 days later they were killed. Half of the rats in which the BPDO was opened were administered L364,718, a CCK-receptor antagonist (0.1 mg/kg/12 hours), 30 minutes before the induction of BPDO. Plasma amylase activity, hematocrit, and pancreatic weight returned to control values after BPDO opening. The highest degree of oxidative stress was found in the pancreases of rats subjected to BPDO for 6 hours, as indicated by the decrease in pancreatic glutathione content, but it was not restored 7 days after BPDO opening. Cell-cycle distribution, as measured with propidium iodide DNA staining, showed increases in the proportion of acinar cells in S-phase from 3 days after BPDO opening in L364,718-treated and nontreated rats. Annexin V-fluorescein isothiocyanate labeling revealed deletion of acinar cells by way of apoptosis 3 days after BPDO opening. However, it may be compensated 7 days after BPDO opening because regardless of whether rats were treated with L364,718, significant increases in synthesis and mitosis were detected. Accumulation of digestive enzymes and calcium in acinar cells was found during BPDO, but this appeared to have normalized 3 days after BPDO opening and onward in both L364,718-treated and nontreated rats. In conclusion, early removal of obstruction allowed rapid cell proliferation and prevented the progression of severe alterations within acinar cells induced by BPDO. CCK blockade does not accelerate pancreatic recovery after BPDO opening.

Topics: Acute Disease; Amylases; Animals; Apoptosis; Calcium; Cell Cycle; Cholecystokinin; Cholestasis, Extrahepatic; Decompression, Surgical; Devazepide; Disease Models, Animal; Flow Cytometry; Hormone Antagonists; Male; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Time Factors; Trypsinogen

Though hereditary pancreatitis is a very rare form of pancreatitis, the discovery of the gene for hereditary pancreatitis provides important implications for pancreatitis in general. A premature activation of trypsinogen is likely to occur not infrequently in our daily life as the onset of the disease is well advanced before a drinking habit starts. It probably goes unnoticed in normal individuals because normal inhibitory mechanisms described above prevent the development of pancreatitis. Any disorders or agents that cause abnormalities in this natural protective mechanism can cause pancreatitis. Genotype-phenotype analyses of CFTR mutations in chronic pancreatitis are necessary to establish the relationship between CFTR and this disease. It remains to be shown how a reduction of functional CFTR causes chronic pancreatitis.

Topics: Acute Disease; Chronic Disease; Cystic Fibrosis; Humans; Pancreatitis; Trypsinogen

The activation peptide released from procarboxypeptidase B, CAPAP, is a marker of the activation of pancreatic enzymes in acute pancreatitis while anionic trypsinogen (AT) levels in urine relate to leakage of unactivated proenzymes. Data on these markers in patients suffering from severe acute abdominal disorders of non-pancreatic origin are lacking.. To examine levels of CAPAP and AT in serum and urine from patients with severe acute abdominal disorders of non-pancreatic origin in order to better define the diagnostic specificity of these two markers in severe acute pancreatitis in relation to other acute intra-abdominal disorders.. The study included 54 patients with severe acute abdominal disorders of non-pancreatic origin with an APACHE II score >3. Immunoreactive CAPAP (irCAPAP) and immunoreactive AT (irAT) were measured in serum and urine using specific immunoassays.. In urine, irCAPAP levels were mildly increased (>2 nmol/l) in 13% of the patients with severe acute abdominal diseases of non-pancreatic origin, but on no occasion did the increase approach the cutoff levels described for severe acute pancreatitis (>100 nmol/l). However, irAT levels in serum and urine were increased (>50 micro g/l) in 54% of the cases.. Contrary to what is found for irAT, patients with acute abdominal pain of non-pancreatic origin rarely have markedly increased levels of irCAPAP in serum and urine.

Topics: Abdominal Pain; Acute Disease; Adult; Aged; Aged, 80 and over; Anions; Biomarkers; Female; Gastrointestinal Diseases; Humans; Male; Middle Aged; Pancreatitis; Peptides; Trypsinogen

Evaluation of a new serum test for diagnosis of acute pancreatitis.. One hundred and sixty-three patients presenting with acute abdominal pain were included into the study. Acute pancreatitis was diagnosed by CT or ultrasound. Serum samples were taken 0-1 days, 2-3 days, and 4-5 days after onset of symptoms and C-reactive protein, lipase, elastase, and amylase were determined. As a further parameter, Pankrin, a newly available kit for the measurement of a mixture of elastase and other pancreatic secretory proteins was used. As control, serum from 558 apparently healthy blood donors was analysed. The receiver operator characteristics (ROC) and the areas under the curves (AUC) were calculated for each individual test.. In Western blot analysis the antibodies of the Pankrin assay detected the majority of protein bands in human pancreatic juice. In blood donors, the median value of Pankrin was 88 U/ml (range 14-316 U/ml). In 16 from 163 patients with acute abdominal pain, acute pancreatitis was diagnosed and the median Pankrin level in samples collected on days 0-1 was 345 U/ml (range 220-518 U/ml, p<0.0001). In those patients with abdominal pain but without pancreatitis, the median was 116 U/ml (range 17-396 U/ml). The ROC-curves for amylase, lipase, elastase, and Pankrin from samples collected after 0-1 days were similar (area under the curves (AUC) >0.98). After 2-3 days, the AUC of all markers decreased (AUC 0.80-0.89) and after 4-5 days the AUC of Pankrin (0.85) was higher than all other parameters.. In those patients with abdominal pain, who present several days after onset of pain, the new serum test for pancreatitis, Pankrin, could be of help to improve the diagnosis of pancreatitis.

Topics: Abdomen, Acute; Abdominal Pain; Amylases; C-Reactive Protein; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Humans; Lipase; Pancreatic Elastase; Pancreatic Juice; Pancreatitis; Predictive Value of Tests; Reagent Kits, Diagnostic; Reference Values; ROC Curve; Sensitivity and Specificity; Trypsinogen

Several genetic factors have been well known to predispose one to chronic pancreatitis (CP). However, little is known about the genetic factors that may provide a protective effect against the disease. Having found a nonsense mutation (c.111C>A; Y37X) and a splicing mutation (IVS2+1G>A) in the cationic trypsinogen gene (protease, serine, 1; PRSS1) in alcoholics without the development of CP, but not in alcoholics with CP and patients with hereditary or idiopathic CP, we propose that while "gain of function" mutations in the PRSS1 gene predispose one to pancreatitis, "loss of function" mutations in the gene may protect one against the disease.

Topics: Adult; Aged; Alcoholism; Chronic Disease; Exons; Female; Genotype; Humans; Male; Middle Aged; Mutation; Pancreatitis; Trypsin; Trypsinogen

1 Kinin B(2) receptor antagonists or tissue kallikrein (t-KK) inhibitors prevent oedema formation and associated sequelae in caerulein-induced pancreatitis in the rat. We have now further investigated the mechanism of kinin generation in the pancreas. 2 Kinins were elevated in the pancreatic tissue already before oedema formation became manifest. Peak values (421+/-59 pmol g(-1) dry wt) were reached at 45 min and remained elevated for at least 2 h; a second increase was observed at 24 h. Pretreatment with the B(2) receptor antagonist icatibant abolished kinin formation, while post-treatment was ineffective. 3 Total kininogen levels were very low in the pancreas of controls, but increased 75-fold during acute pancreatitis. This increase was absent in rats that were pretreated with icatibant. 4 During pancreatitis, t-KK-like and plasma kallikrein (p-KK)-like activity in the pancreas, as well as trypsinogen activation peptide (TAP) increased significantly. Icatibant pretreatment further augmented t-KK about 100-fold, while p-KK was significantly attenuated; TAP levels remained unaffected. 5 Endogenous protease inhibitors (alpha(1)-antitrypsin, alpha(2)-macroglobulin) were low in normal tissues, but increased 45- and four-fold, respectively, during pancreatitis. This increase was abolished when oedema formation was prevented by icatibant. 6 In summary, oedema formation is initiated by t-KK; the ensuing plasma protein extravasation supplies further kininogen and active p-KK to the tissue. Concomitantly, endogenous protease inhibitors in the oedema fluid inhibit up to 99% of active t-KK. Our data thus suggest a complex interaction between kinin action and kinin generation involving positive and negative feedback actions of the inflammatory oedema.

Topics: Acute Disease; alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Anti-Inflammatory Agents, Non-Steroidal; Bradykinin; Ceruletide; Edema; Enzyme Activation; Female; Kininogens; Kinins; Pancreas; Pancreatitis; Plasma Kallikrein; Rats; Rats, Sprague-Dawley; Receptors, Cell Surface; Serine Proteinase Inhibitors; Tissue Kallikreins; Trypsinogen

Topics: Acute Disease; Diagnosis, Differential; Electrocardiography; Genetic Predisposition to Disease; Humans; Lipase; Mutation; Myocardial Infarction; Pancreatitis; Prognosis; Trypsinogen

Trypsinogen activation within acinar cells plays a crucial role in the pathogenesis of acute pancreatitis (AP). Our aim was to characterize temporal changes of trypsinogen-1, trypsinogen-2, complexes of trypsin-1-alpha1-antitrypsin (T1-AAT) and trypsin-2-alpha1-antitrypsin (T2-AAT), trypsinogen activation peptide (TAP) and pancreatic secretory trypsin inhibitor (PSTI) in patients with AP.. The study comprised 64 consecutive patients with AP (19 with severe disease) and 32 controls. The concentrations of trypsinogen-1 and -2, PSTI, T1-AAT and T2-AAT were determined by time-resolved immunofluorometric assays (IFMA), and TAP was measured using a competitive enzyme immunoassay from serum and urine.. The concentrations of trypsinogen-1 and -2 in serum reflected similar patterns, but excretion of trypsinogen-1 into urine was markedly lower than that of trypsinogen-2, the concentration of which correlated strongly with disease severity. The concentrations of T1-AAT were no higher in severe AP than in mild AP, while T2-AAT concentrations were significantly higher in severe than in mild disease. PSTI increased over the course of several days, showing strong correlation with disease severity. The concentrations of plasma and urinary TAP decreased rapidly to undetectable levels. During the early phase of AP, TAP correlated with the disease severity in plasma and urine but there was no difference between controls and patients with mild AP.. More pronounced changes in trypsinogen-2 and its complex with AAT than in those of trypsinogen-1 were demonstrated, suggesting that trypsinogen-2 might play a more important role in the pathogenesis of AP than earlier believed. Urinary PSTI showed features warranting further investigations as a marker of disease severity.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin; Biomarkers; Carrier Proteins; Female; Growth Substances; Humans; Intercellular Signaling Peptides and Proteins; Male; Middle Aged; Oligopeptides; Pancreatitis; Time Factors; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

There is an obvious clinical need for a simple test that can identify patients at risk of developing severe acute pancreatitis. In this work we compared urinary trypsinogen-2 with urinary trypsinogen activation peptide (TAP) and serum C-reactive protein (CRP) for early differentiation between mild and severe acute pancreatitis.. The study population consisted of 127 consecutive patients with acute pancreatitis of whom 29 had severe disease. Urinary trypsinogen-2 was measured by a quantitative immunofluorometric assay and TAP by a competitive immunoassay. Serum CRP was determined by immunoturbidimetry.. The sensitivity and specificity to identify severe acute pancreatitis on admission was 72% and 81% for urinary trypsinogen-2, 64% and 82% for urinary TAP, and 29% and 93% for serum CRP, respectively. At 24 h after admission, the values were 82% and 78% for urinary trypsinogen-2, 52% and 92% for urinary TAP, and 84% and 72% for serum CRP, respectively. Receiver-operating characteristics curve analysis showed that the area under the curve was larger for urinary trypsinogen-2 than for urinary TAP and serum CRP on admission and 24 h after admission. On admission the positive likelihood ration for urinary trypsiongen-2 was 3.7, for urinary TAP 3.6, and 4.3 for serum CRP, respectively. The corresponding negative likelihood ratios were 0.34, 0.43, and 0.76, respectively.. Urinary trypsinogen-2 was superior to serum CRP and as god as or even better than urinary TAP and in the early prediction of disease severity in acute pancreatitis. These results suggest that it could be a valuable adjunct in the early assessment of the severity of acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; APACHE; C-Reactive Protein; Diagnosis, Differential; Female; Humans; Male; Middle Aged; Oligopeptides; Pancreatitis; Pancreatitis, Alcoholic; Prospective Studies; Reference Values; ROC Curve; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen

The clinical course of chronic pancreatitis in patients with mutations of cationic trypsinogen and the trypsin inhibitor SPINK1 has not yet been characterized.. Cationic trypsinogen (PRSS1) and the serine protease inhibitor, Kazal type 1 (SPINK1), were analyzed in patients with pancreatitis of unclear origin.. Eighty subjects with trypsinogen mutations (21x N29I, 59x R122H) and 59 patients with the SPINK1 N34S variant (11 homozygous, 48 heterozygous) were included in the study.. In patients with mutations of PRSS1 (N29I, R122H) and SPINK1 (N34S) the parameters such as calcification, dilatation of the main pancreatic duct, diabetes mellitus, hospital treatments, and surgery were recorded.. Case control studies were performed to compare both mutational groups, and the follow-up time served as a matching criterion. The Kaplan-Meier analysis was used to estimate the time course of the symptoms.. Ten years after the onset of the disease, the probability (+/-SE) of symptoms in patients with PRSS1 mutations was as follows: 1st hospital stay: 86+/-4%; calcification: 21+/-4%; duct dilatation: 26+/-9%; surgery: 19+/-5%; diabetes: 6+/-5%. After 25 years, we found the following data: 1st hospital stay: 96+/-3%; calcification: 38+/-8%; duct dilatation: 38+/-8%; surgery: 37+/-10%; diabetes: 28+/-8%. A case-control-study of 38 pairs of patients with either PRSS1 or SPINK1 mutations showed that the probability of duct dilatation, diabetes and calcification was slightly higher in patients having a SPINK1 mutation. There was no difference between those subjects with a homozygous or heterozygous SPINK1 mutation. In comparison to alcoholic chronic pancreatitis patients, the PRSS1 associated disease revealed a lower frequency of calcification and diabetes.. The progression of chronic pancreatitis was slightly more rapid in patients with SPINK1 mutations than in patients with cationic trypsinogen mutations, but was much less than in those having alcoholic chronic pancreatitis.

Topics: Case-Control Studies; Child; Chronic Disease; Disease Progression; Follow-Up Studies; Genetic Predisposition to Disease; Humans; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Acute Disease; Adult; Base Sequence; DNA; DNA Mutational Analysis; Female; Humans; Mutation, Missense; Pancreatitis; Trypsin; Trypsinogen

Mutations in cystic fibrosis transmembrane conductance regulator (CFTR), in cationic trypsinogen (PRSS1) and in serine protease inhibitor Kazal type 1 (SPINK1) genes have been associated with chronic pancreatitis (alcohol related, idiopathic and hereditary). However, the inheritance pattern is still not clear.. Eighty-two unrelated Brazilian patients with chronic pancreatitis (alcohol-related disease in 64, idiopathic disease in 16, and hereditary disease in 2). Two hundred unrelated individuals with an ethnic distribution comparable to the patients were studied as controls.. Detection of mutations in CFTR, PRSS1, and SPINK1 genes.. Mutations in the CFTR gene were found in 8 patients (9.8%) with chronic pancreatitis, 5 of them with idiopathic disease. Interestingly, the only clinical symptom in a male patient in the alcoholic group, who was a compound heterozygote (DeltaF508/R170C) for two CFTR mutations, was pancreatitis without infertility or pulmonary involvement. In the PRSS1 gene, the E79K change in exon 3 was found in one patient (1.2%) with alcohol-related chronic pancreatitis. Four different alterations were identified in the SPINK1 gene.. Mutations in the CFTR gene represent the major cause of idiopathic chronic pancreatitis in Brazilian patients. No mutation was found in the PRSS1 gene among our patients suggesting further genetic heterogeneity for hereditary and idiopathic chronic pancreatitis. Interestingly, the most frequent SPINK1 N34S mutation was not present in patients or controls. Moreover, the -253C allele for the SPINK1 gene was significantly more frequent in patients than controls (P=0.004), suggesting that it might represent a risk factor for the development of pancreatitis in our population.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Brazil; Child; Chronic Disease; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Female; Genetic Predisposition to Disease; Genetic Testing; Humans; Male; Middle Aged; Mutation; Pancreatitis; Pancreatitis, Alcoholic; Serine Proteinase Inhibitors; Trypsin; Trypsinogen

The role of potent vasoconstrictor endothelin-1 (ET-1) in acute pancreatitis (AP) remains controversial. The aim was to compare the effect of nonselective ET RA/B (LU-302872) and selective ET RA (LU-302146) antagonists on pancreatic histology, ultrastructure and trypsinogen activation in severe taurocholate AP in rats. Male Wistar rats with AP were treated with an intraperitoneous injection of 1, 5 and 10 mg/kg of body weight of each antagonist at 0, 6, 12 and 18 h after taurocholate administration. After 24 h, the samples of pancreases were taken for histological and ultrastructural examinations and for assessment of trypsinogen activation. Both antagonists, at all investigated doses, decreased the damage to the acinar cells detected in the light microscope and ultrastructurally. Trypsinogen activation increased to 29.7 +/- 3.9% in the AP untreated in comparison to the control group [12.7 +/- 1.4% (P<0.001)]. This increase was attenuated to 13.8 +/- 2.2% in AP treated with a high dose of the nonselective antagonist and to 8.4 +/- 1.7% with low dose of selective antagonist. The obtained results indicate that ET-1 could participate in the damage to the pancreas during AP. Both antagonists of ET-1 receptors exerted a similar beneficial effect on the morphological changes of the pancreas in AP. One of the probable mechanism could be the attenuation of trypsinogen activation.

Topics: Acute Disease; Animals; Benzhydryl Compounds; Endothelin Receptor Antagonists; Male; Microscopy, Electron; Pancreas; Pancreatitis; Propionates; Pyrimidines; Rats; Rats, Wistar; Receptors, Endothelin; Taurocholic Acid; Trypsinogen

Recent genetic investigations into cationic trypsinogen and pancreatic secretory trypsin inhibitor (PSTI) led to the conclusion that mutations in either gene can contribute to the development of (hereditary) chronic pancreatitis. Since genetic animal models are not available yet, we have studied the Wistar-Bonn/Kobori (WBN/Kob) rat, a model for chronic pancreatitis (CP). To explore the possibility that PSTI may be secreted at lower levels or contain a mutation in the WBN/Kob rat, we investigated the masses of PSTI-I and -II and asked whether the ratio of PSTI/trypsinogen is decreased in animals with CP.. We collected pancreatic juice from WBN/Kob and Wistar rats aged 6-36 weeks and measured PSTI-I (ELISA) and trypsin.. PSTI-I and -II were identified in Wistar and WBN/Kob rats by mass spectrometry and N-terminal sequencing. Using a newly developed PSTI-I ELISA, we can show that the PSTI-I/trypsinogen ratio is not decreased but rather increased in WBN/Kob rats compared to healthy Wistar rats. No evidence for a PSTI mutation was found.. Our data does not support the hypothesis that a dysbalance of PSTI/trypsinogen ratio is a causative factor for CP.

Topics: Animals; Chronic Disease; Disease Models, Animal; Male; Pancreatitis; Protein Isoforms; Rats; Rats, Wistar; Reference Values; RNA, Messenger; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Heat shock proteins (HSPs) are necessary in the synthesis, degradation, folding, transport, and translocation of different proteins. It is well known that the increased expression of HSPs may have a protective effect against cerulein-induced pancreatitis in rats or against choline-deficient ethionine-supplemented diet model pancreatitis in mice. The aim of this study was to investigate the potential effects of HSP preinduction by cold or hot water immersion on trypsin-induced acute pancreatitis in rats. Trypsin was injected into the interlobular tissue of the duodenal part of the pancreas at the peak level of HSP synthesis, as determined by Western blot analysis. The rats were sacrificed by exsanguination through the abdominal aorta 6 h after the trypsin injection. The serum amylase activity, the tumor necrosis factor-alpha, interleukin-1, and interleukin-6 levels, the pancreatic weight/body weight ratio, and the pancreatic contents of DNA, protein, amylase, lipase, and trypsinogen were measured. A biopsy for histology was taken. Hot water immersion significantly elevated the HSP72 expression, while cold water immersion significantly increased the HSP60 expression. Cold water immersion pretreatment ameliorated the pancreatic edema in trypsin-induced pancreatitis, however this was not due to the HSP60. Hot water immersion pretreatment did not have any effect on the measured parameters in trypsin-induced pancreatitis. The findings suggest that the induction of HSP60 or HSP72 are not enough to protect rats against the early phase of this localized necrohemorrhagic pancreatitis model.

Topics: Amylases; Animals; Antibodies; Antibody Specificity; Blotting, Western; Body Weight; Chaperonin 60; Cold Temperature; Cytokines; Disease Models, Animal; DNA; Heat-Shock Proteins; Hot Temperature; HSP72 Heat-Shock Proteins; Immersion; Lipase; Male; Organ Size; Pancreas; Pancreatitis; Proteins; Rats; Rats, Wistar; Stress, Physiological; Trypsin; Trypsinogen

Intrapancreatic activation of trypsinogen is believed to play a critical role in the initiation of acute pancreatitis, but mechanisms responsible for intrapancreatic trypsinogen activation during pancreatitis have not been clearly defined. In previous in vitro studies, we have shown that intra-acinar cell activation of trypsinogen and acinar cell injury in response to supramaximal secretagogue stimulation could be prevented by the cell permeant cathepsin B inhibitor E64d (Saluja A, Donovan EA, Yamanaka K, Yamaguchi Y, Hofbauer B, and Steer ML. Gastroenterology 113: 304-310, 1997). The present studies evaluated the role of intrapancreatic trypsinogen activation, this time under in vivo conditions, in two models of pancreatitis by using another highly soluble cell permeant cathepsin B inhibitor, L-3-trans-(propylcarbamoyl)oxirane-2-carbonyl-L-isoleucyl-L-proline methyl ester (CA-074me). Intravenous administration of CA-074me (10 mg/kg) before induction of either secretagogue-elicited pancreatitis in mice or duct infusion-elicited pancreatitis in rats markedly reduced the extent of intrapancreatic trypsinogen activation and substantially reduced the severity of both pancreatitis models. These observations support the hypothesis that, during the early stages of pancreatitis, trypsinogen activation in the pancreas is mediated by the lysosomal enzyme cathepsin B. Our findings also suggest that pharmacological interventions that inhibit cathepsin B may prove useful in preventing acute pancreatitis or reducing its severity.

Topics: Animals; Cathepsin B; Ceruletide; Cysteine Proteinase Inhibitors; Dipeptides; Infusions, Parenteral; Male; Mice; Mice, Inbred Strains; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Severity of Illness Index; Taurocholic Acid; Trypsinogen

The aim of this study was to examine the effect of the most potent CCK receptor antagonist, L364,718, on two major factors involved in pancreatitis development: enzyme load and cytosolic calcium (Ca2+) levels in acinar cells. L364,718 (0.1 mg/kg/12 hr) was administered from 30 min before inducing acute pancreatitis (AP) by pancreatic duct obstruction (PDO) for 48 hr. The results obtained at different AP stages in PDO rats treated and not treated with the CCK antagonist were compared. Similar increases in the intracellular enzyme content were found at earlier stages of pancreatitis in all PDO rats treated or not treated with L364,718. The CCK antagonist increased cytosolic Ca2+ levels up to 6 hr after administration, inducing a higher cytosolic Ca2+ overload at the earliest stages of pancreatitis in L364,718-treated PDO rats than in those not treated. This event might justify the higher increases in ascites volume and haematocrit found in PDO rats treated with L364,718 and the exacerbation in pancreatic morphological alterations induced by PDO. The CCK receptor antagonist L364,718 produces alterations in the acinar calcium homeostasis that prevent to reduction in the severity of pancreatitis induced by obstruction.

Topics: Acute Disease; Amylases; Animals; Calcium; Cholecystokinin; Cholestasis; Devazepide; Hormone Antagonists; Male; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

Topics: Acute Disease; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Humans; Pancreatic Neoplasms; Pancreatitis; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Topics: Adolescent; Chronic Disease; Female; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

Tropical calcific pancreatitis (TCP) is a chronic pancreatitis unique to developing countries in tropical regions. The cause of TCP is obscure. Whereas environmental factors, such as protein energy malnutrition and ingestion of cassava, have been implicated, a genetic predisposition to the disease also may be important. In the present study we report on mutations in the serine protease inhibitor, Kazal type 1 (SPINK1) gene in north Indian patients with TCP.. We studied 66 unrelated TCP patients (44 men, 49 with diabetes, and 6 with family history of TCP), 25 relatives, and 92 healthy control subjects. Samples were analyzed for SPINK1 variants (-53C>T, L14P, N34S, P55S, and 272T>C) and cationic trypsinogen (PRSS1) variants (A16V, K23R, N29I, and R122H) by melting curve analysis.. Twenty-nine patients (44%) carried the N34S missense mutation, of whom 9 (14%) were homozygotes. In contrast, only 2 (2.2%) control subjects were N34S heterozygotes (prevalence ratio 20.2; 95% confidence interval 5.0-81.8; P < 0.0001 vs. TCP). The severity of pancreatitis did not differ between TCP patients with or without N34S, or among those heterozygous or homozygous for N34S. Among TCP patients with or without diabetes, the frequency of N34S carriers (43% vs. 47%) and N34S homozygotes (14% vs. 12%) was similar.. TCP is highly associated with the SPINK1 N34S mutation. The high prevalence of N34S in TCP patients with and without diabetes suggests that these 2 subtypes have a similar genetic predisposition. The genetic predisposition to TCP resembles, at least in part, the idiopathic chronic pancreatitis found in industrialized countries.

Topics: Adult; Chronic Disease; Diabetes Mellitus; Family Health; Female; Heterozygote; Homozygote; Humans; India; Male; Pancreatitis; Pedigree; Point Mutation; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Cyclooxygenase-2 (COX-2), a widely distributed enzyme, plays an important role in inflammation. We have studied the role of COX-2 in acute pancreatitis and pancreatitis-associated lung injury using both the pharmacological inhibition of COX-2 and genetic deletion of COX-2. Pancreatitis was induced in mice by 12 hourly injections of cerulein. The severity of pancreatitis was assessed by measuring serum amylase, pancreatic trypsin activity, intrapancreatic sequestration of neutrophils, and acinar cell necrosis. The severity of lung injury was evaluated by measuring lactate dehydrogenase levels in the bronchoalveolar lavage fluid and by quantitating neutrophil sequestration in the lung. In both the pharmacologically inhibited and genetically altered mice, the severity of pancreatitis and pancreatitis-associated lung injury was reduced compared with the noninhibited strains of COX-2-sufficient mice. This reduction in injury indicates that COX-2 plays an important proinflammatory role in pancreatitis and its associated lung injury. Our findings support the concept that COX-2 inhibitors may play a beneficial role in the prevention of acute pancreatitis or in the reduction of its severity.

Topics: Animals; Celecoxib; Ceruletide; Cyclooxygenase 2; Cyclooxygenase 2 Inhibitors; Cyclooxygenase Inhibitors; HSP70 Heat-Shock Proteins; Isoenzymes; Lung Diseases; Mice; Mice, Inbred C57BL; Mice, Knockout; NF-kappa B; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; Nitrobenzenes; Pancreatitis; Prostaglandin-Endoperoxide Synthases; Pyrazoles; RNA, Messenger; Severity of Illness Index; Sulfonamides; Trypsinogen

Rat P23 is an isoform of trypsin (ogens) synthesized by rat acinar cells. Expression of P23 is stimulated strongly by caerulein, an analogue of cholecystokinin (CCK). However, the physiological relevance of rat P23 in healthy and pathological conditions such as caerulein-induced pancreatitis is largely unknown. Using recombinant P23 trypsinogen and reconstitution analysis of zymogen autoactivation, unique inhibitor-resistance characteristics of P23 were elucidated. P23 cDNA was expressed in Escherichia coli periplasm, yielding recombinant P23 trypsinogen. Autoactivation of zymogen granule contents from caerulein-induced rat pancreas was also studied. Activation kinetics of P23 by enterokinase was similar to those of rat anionic trypsinogen, which is a major isoform of trypsinogen. Interestingly, rat pancreatic secretory trypsin inhibitor (PSTI), which protects against deleterious activation of trypsinogens in zymogen granules, failed to inhibit P23 trypsin even with four-fold molar excess, at which concentration it effectively inhibited rat anionic trypsin to almost 100%. P23 trypsin also showed marked resistance to proteinaceous trypsin inhibitors such as soybean trypsin inhibitor and aprotinin. P23 trypsin activated by enterokinase dramatically accelerated the cascade of autoactivation of anionic trypsinogen even in the presence of PSTI. Taken together with a previous observation that P23 is specifically upregulated 14-fold by 24-h caerulein infusion, these results suggest that elevated levels of P23 should be taken into consideration in the mechanism of trypsinogens within the pancreas in pathological conditions.

Topics: Amino Acid Sequence; Animals; Ceruletide; Cloning, Molecular; Enteropeptidase; Enzyme Activation; Escherichia coli; Pancreas; Pancreatitis; Protein Isoforms; Rats; Trypsin Inhibitors; Trypsinogen

Topics: Adolescent; Adult; Age of Onset; Child; Chronic Disease; Humans; Mutation; Pancreatitis; Trypsin; Trypsinogen

Topics: Chlorides; DNA Mutational Analysis; Genetic Predisposition to Disease; Humans; Membrane Transport Proteins; Pancreatitis; Risk; Serine Proteinase Inhibitors; Trypsin; Trypsinogen

Enzyme load in pancreas has been considered a risk factor in the development of acute pancreatitis. In order to confirm this hypothesis our aim was to analyze the development and evolution of acute pancreatitis (AP) induced by bile-pancreatic duct obstruction (BPDO) after reducing the pancreatic enzyme content. L-364,718 - a potent CCK-receptor antagonist - was administered (0.1 mg/kg/day) for 7 days before inducing AP by BPDO. The course of AP was evaluated at different times from 1.5-48 h after BPDO. Amylase and trypsinogen contents and cytosolic calcium levels were measured by flow cytometry using specific antisera against pancreatic enzymes labelled with isothiocyanate of fluorescein and Fluo 3, respectively. The severity of the disease at the different stages was evaluated by measurements of amylase activity in ascites and plasma, percentage of pancreatic fluid and haematocrit. Electron microscopy study of the pancreas showed an increased number of zymogen granules spread through the acinar cells of control rats treated with L-364,718 for 7 days, however, total enzyme content in individual acinar cells was significantly (p < 0.01) diminished. AP significantly increased intracellular amylase and trypsinogen load from 3-12 h after BPDO, and prior L-364,718 treatment enhanced the blockade of enzyme secretion. As a result, acinar enzyme content was significantly increased from earlier stages (1.5 h after BPDO). In parallel, increased cytosolic calcium levels observed up to 24 h after BPDO appeared earlier in L-364,718-treated rats than in those not treated. The severity of AP seems to have been higher in rats previously treated with the CCK-receptor antagonist as indicated by the significantly higher pancreatic fluid and amylase activity in ascites and plasma observed at different times after BPDO. Our results indicate that there is no correlation between the severity of pancreatitis and the amount of enzymes accumulated in the pancreas before the disease is induced.

Topics: Acute Disease; Amylases; Animals; Bile Ducts; Calcium; Cholestasis, Extrahepatic; Devazepide; Disease Models, Animal; Male; Microscopy, Electron; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Severity of Illness Index; Time Factors; Trypsinogen

Hereditary pancreatitis has been found to be associated with germline mutations in the cationic trypsinogen (PRSS1) gene. Here we report a family with hereditary pancreatitis that carries a novel PRSS1 mutation (R122C). This mutation cannot be diagnosed with the conventional screening method using AflIII restriction enzyme digest. We therefore propose a new assay based on restriction enzyme digest with BstUI, a technique that permits detection of the novel R122C mutation in addition to the most common R122H mutation, and even in the presence of a recently reported neutral polymorphism that prevents its detection by the AflIII method. Recombinantly expressed R122C mutant human trypsinogen was found to undergo greatly reduced autoactivation and cathepsin B-induced activation, which is most likely caused by misfolding or disulfide mismatches of the mutant zymogen. The K(m) of R122C trypsin was found to be unchanged, but its k(cat) was reduced to 37% of the wild type. After correction for enterokinase activatable activity, and specifically in the absence of calcium, the R122C mutant was more resistant to autolysis than the wild type and autoactivated more rapidly at pH 8. Molecular modeling of the R122C mutant trypsin predicted an unimpaired active site but an altered stability of the calcium binding loop. This previously unknown trypsinogen mutation is associated with hereditary pancreatitis, requires a novel diagnostic screening method, and, for the first time, raises the question whether a gain or a loss of trypsin function participates in the onset of pancreatitis.

Topics: Amino Acid Sequence; Arginine; Binding Sites; Calcium; Cysteine; Deoxyribonucleases, Type II Site-Specific; Female; Humans; Kinetics; Male; Models, Molecular; Molecular Sequence Data; Mutation; Pancreatitis; Pedigree; Phosphorylation; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Protein Binding; Protein Structure, Tertiary; Recombinant Proteins; Sequence Analysis, DNA; Time Factors; Trypsin; Trypsinogen

Heat shock proteins (HSPs) have indispensable functions in the synthesis, degradation, folding, transport, and translocation of intracellular proteins. HSPs are proteins that help cells to survive stress conditions by repairing damaged proteins.. To investigate the potential effects of HSP preinduction by cold-water (CWI) or hot-water immersion (HWI) on sodium taurocholate (TC)-induced acute pancreatitis in rats.. TC was injected into the common biliopancreatic duct of the animals at the peak level of HSP synthesis, as determined by Western blot analysis. The rats were killed by exsanguination through the abdominal aorta 6 hours after the TC injection. The serum amylase activity, the IL-1, IL-6 and TNF-alpha levels, the pancreatic weight/body weight ratio, and the pancreatic contents of DNA, protein, amylase, lipase, and trypsinogen were measured, and a biopsy for histology was taken.. HWI significantly elevated HSP72 expression, whereas CWI significantly increased HSP60 expression. It was demonstrated that CWI pretreatment ameliorated the pancreatic edema and the serum amylase level increase, whereas the morphologic damage was more severe in this form of acute pancreatitis. HWI pretreatment did not have any effects on the measured parameters in TC-induced pancreatitis.. The findings suggest a possible role of HSP60, but not HSP72, in the slight protection in the early phase of this necrohemorrhagic pancreatitis model.

Topics: Amylases; Animals; Body Weight; Chaperonin 60; Cytokines; Disease Models, Animal; DNA; Fever; Heat-Shock Proteins; HSP72 Heat-Shock Proteins; Hypothermia; Lipase; Male; Organ Size; Pancreas; Pancreatitis; Rats; Rats, Wistar; Taurocholic Acid; Trypsinogen

Mutation of Arg(117), an autocatalytic cleavage site, is the most frequent amino acid change found in the cationic trypsinogen (Tg) of patients with hereditary pancreatitis. In the present study, the role of Arg(117) was investigated in wild-type cationic Tg and in the activation-resistant Lys(15) --> Gln mutant (K15Q-Tg), in which Tg-specific properties of Arg(117) can be examined selectively. We found that trypsinolytic cleavage of the Arg(117)-Val(118) bond did not proceed to completion, but due to trypsin-catalyzed re-synthesis an equilibrium was established between intact Tg and its cleaved, two-chain form. In the absence of Ca(2+), at pH 8.0, the hydrolysis equilibrium (K(hyd) = [cleaved Tg]/[intact Tg]) was 5.4, whereas 5 mm Ca(2+) reduced the rate of cleavage at Arg(117) at least 20-fold, and shifted K(hyd) to 0.7. These observations indicate that the Arg(117)-Val(118) bond exhibits properties analogous to the reactive site bond of canonical trypsin inhibitors and suggest that this surface loop might serve as a low affinity inhibitor of zymogen activation. Consistent with this notion, autoactivation of cationic Tg was inhibited by the cleaved form of K15Q-Tg, with an estimated K(i) of 80 microm, while no inhibition was observed with K15Q-Tg carrying the Arg(117) --> His mutation. Finally, zymogen breakdown due to other trypsinolytic pathways was shown to proceed almost 2000-fold slower than cleavage at Arg(117). Taken together, the findings suggest two independent, successively functional trypsin-mediated mechanisms against pathological Tg activation in the pancreas. At low trypsin concentrations, cleavage at Arg(117) results in inhibition of trypsin, whereas high trypsin concentrations degrade Tg, thus limiting further zymogen activation. Loss of Arg(117)-dependent trypsin inhibition can contribute to the development of hereditary pancreatitis associated with the Arg(117) --> His mutation.

Topics: Amino Acid Substitution; Arginine; Binding Sites; Dimerization; Enzyme Precursors; Humans; Kinetics; Mutagenesis, Site-Directed; Mutation, Missense; Pancreatitis; Trypsinogen

Recent studies have indicated that prior thermal stress causes upregulation of heat shock protein 70 (HSP70) expression in the pancreas and protects against secretagogue induced pancreatitis. The mechanisms responsible for the protective effect are not known. Similarly, the effects of prior non-thermal stress on HSP70 expression and pancreatitis are not known. The current studies were designed to specifically address these issues.. In the current studies pancreatitis was induced by administration of a supramaximally stimulating dose of caerulein 12 hours after thermal stress and 24 hours after non-thermal (that is, beta adrenergic stimulation) stress.. Both thermal and non-thermal stresses caused pancreatic HSP70 levels to rise and resulted in increased expression of HSP70 in acinar cells. Both forms of stresses protected against caerulein induced pancreatitis and prevented the early intrapancreatic activation of trypsinogen which occurs in this model of pancreatitis.. These results suggest that both thermal and non-thermal stresses protect against pancreatitis by preventing intrapancreatic digestive enzyme activation and that HSP70 may mediate this protective effect.

Topics: Amylases; Analysis of Variance; Animals; Blotting, Western; Ceruletide; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; HSP70 Heat-Shock Proteins; Hyperthermia, Induced; Luminescent Measurements; Male; Pancreatitis; Peroxidase; Rats; Rats, Wistar; Stress, Physiological; Trypsinogen

Prior thermal stress induces heat shock protein 70 (HSP70) expression in the pancreas and protects against secretagogue-induced pancreatitis, but it is not clear that this thermal stress-induced protection is actually mediated by HSP70 since thermal stress may have other, non-HSP related, effects.. In the present study, we have administered antisense (AS) oligonucleotides, which prevent pancreatic expression of HSP70 to rats, in vivo, to evaluate this issue. In a separate series of experiments, designed to examine the role of pancreatitis-induced HSP70 expression in modulating the severity of pancreatitis, rats not subjected to prior thermal stress were given AS-HSP70 before cerulein administration, and trypsinogen activation as well as the severity of pancreatitis were evaluated.. Hyperthermia induced HSP70 expression, prevented intrapancreatic trypsinogen activation, and protected against cerulein-induced pancreatitis. Administration of AS-HSP70 but not sense-HSP70 reduced the thermal stress-induced HSP70 expression, restored the ability of supramaximal cerulein stimulation to cause intrapancreatic trypsinogen activation, and abolished the protective effect of prior thermal stress against pancreatitis. In non-thermally stressed animals, pretreatment with AS-HSP70 before the induction of pancreatitis exacerbated all the parameters associated with pancreatitis.. These findings lead us to conclude that HSP70 induction, rather than some other thermal stress-related phenomenon, mediates the thermal stress-induced protection against pancreatitis and that it protects against pancreatitis by preventing intrapancreatic activation of trypsinogen. The worsening of pancreatitis, which occurs when non-thermally stressed animals are given AS-HSP70 before cerulein, suggests that cerulein-induced HSP70 expression in nontreated animals acts to limit the severity of pancreatitis.

Topics: Acute Disease; Animals; Ceruletide; Gene Expression Regulation; Heat Stress Disorders; HSP70 Heat-Shock Proteins; Oligonucleotides, Antisense; Pancreatitis; Rats; Rats, Wistar; Stress, Physiological; Trypsinogen

Hereditary pancreatitis (HP) is usually caused by mutations in the cationic trypsinogen (PRSS1) gene, especially R122H or N29I. We sequenced the PRSS1 gene in the proband of families without these common mutations. Novel R122C and N29T mutations were detected in independent families that segregated with the disease in an autosomal dominant fashion. The R122C mutation eliminates the arginine autolysis site as with R122H mutations. The N29T mutation may also enhance intrapancreatic trypsin activity as has been demonstrated in vitro. Identification of these new mutations requires special attention as commonly used detection methods may fail.

Topics: Adult; Amino Acid Substitution; Chronic Disease; Female; Humans; Male; Mutation; Pancreatitis; Pedigree; Phenotype; Polymorphism, Restriction Fragment Length; Trypsinogen

A premature and intracellular activation of digestive zymogens is thought to be responsible for the onset of pancreatitis. Because trypsin has a critical role in initiating the activation cascade of digestive enzymes in the gut, it has been assumed that trypsin also initiates intracellular zymogen activation in the pancreas. We have tested this hypothesis in isolated acini and lobules from rat pancreas. Intracellular trypsinogen activation was induced by supramaximal secretagogue stimulation and measured using either specific trypsin substrates or immunoreactivity of the trypsinogen activation peptide (TAP). To prevent a trypsin-induced trypsinogen activation, we used the cell-permeant, highly specific, and reversible inhibitor Nalpha-(2-naphthylsulfonyl)-3-amidinophenylalanine-carboxymethylpiperazide (S124), and to prevent cathepsin-induced trypsinogen activation, we used the cysteine protease inhibitor E-64d. Incubation of acini or lobules in the presence of S124 completely prevented the generation of trypsin activity in response to supramaximal caerulein but had no effect whatsoever on the generation of TAP. Conversely, when trypsin activity was recovered at the end of the experiment by either washout of S124 from acini or extensive dilution of lobule homogenates, it was up to 400% higher than after caerulein alone and corresponded, in molar terms, to the generation of TAP. Both trypsin activity and TAP release were inhibited in parallel by E-64d. We conclude that caerulein-induced trypsinogen activation in the pancreas is caused by an E-64d-inhibitable mechanism such as cathepsin-induced trypsinogen activation, and neither involves nor requires intracellular trypsin activity. Specific trypsin inhibition, on the other hand, prevents 80% of trypsin inactivation or autodegradation in the pancreas.

Topics: Acute Disease; Animals; Cathepsin B; Ceruletide; Enzyme Activation; Male; Naphthalenes; Pancreas; Pancreatitis; Piperazines; Rats; Rats, Wistar; Trypsin; Trypsin Inhibitors; Trypsinogen

The role of nuclear factor kappaB (NF-kappaB) activation in acute pancreatitis is uncertain. The transcription factor NF-kappaB is activated early in acute pancreatitis, and NF-kappaB is widely considered a key element in inflammatory responses based on its ability to regulate the expression of inflammatory mediators in vitro. However, its role in vivo in specific diseases remains unclear, and the current data on the role of NF-kappaB in acute pancreatitis is primarily correlative.. In this study, NF-kappaB was directly activated within the pancreas using adenoviral-mediated transfer of an active subunit, RelA/p65 (Adp65), delivered by intraductal injection.. Administration of Adp65 led to the infection of a population of acinar cells within the pancreas, the activation of NF-kappaB, the expression of NF-kappaB target genes, and an inflammatory response. Administration of Adp65 increased the infiltration of neutrophils to the pancreas and lung and caused widespread damage to pancreatic acinar cells. In contrast, at the same titer, control adenovirus (AdGFP) had no effect on these parameters. The level of NF-kappaB activation and the severity of inflammation were reduced when an adenovirus bearing the inhibitory subunit IkappaB-alpha was coadministered with Adp65.. Thus, activation of NF-kappaB within the pancreas was sufficient for the initiation of an inflammatory response in this model. These results help define the specific role of NF-kappaB activation in acute pancreatitis.

Topics: Acute Disease; Adenoviridae; Animals; Gene Expression; Genetic Vectors; Male; Neutrophils; NF-kappa B; Pancreas; Pancreatitis; Rats; Rats, Wistar; RNA, Messenger; Transcription Factor RelA; Trypsinogen

Mutations of the cationic trypsinogen (CT) and the serine protease inhibitor, Kazal type 1 (SPINK 1) are associated with chronic pancreatitis. After mutational screening of a cohort of patients with nonalcoholic chronic pancreatitis, we report three novel variants of the trypsinogen molecule and the clinical characteristics of the carriers.. The coding region of the exon 2 and 3 of the CT gene of 523 patients with chronic nonalcoholic pancreatitis (108 patients with suspected hereditary pancreatitis (HP) and 415 patients with "idio pathic" pancreatitis [IP]) and 82 controls was analyzed after polymerase chain reaction amplification. Clinical characteristics were obtained by questioning the patients and their relatives and physicians. HP was suspected when two members of a family had chronic pancreatitis. A restriction digestion was used to analyze the N34S mutation SPINK1.. The mutation R122H of the cationic trypsinogen was found in 21 index patients, N291 in six index patients, and A16V and D22G in one index patient, all from HP families. The N34S mutation of SPINK1 was found in two index patients with a family history of HP. In three patients, the novel point mutations L104P, R116C, and C139F of the cationic trypsinogen were found. A clear autosomally dominant inheritance of chronic pancreatitis was not present in these families. In 75 index patients from HP families (69.4%), no mutation could be found. The SPINK 1-mutation N34S was detected in only one patient carrying a CT mutation, and was found in 68 (16.4%) of patients with IP.. The R122H and N291 mutations of CT are the most common disease-associated mutations in HP; the N34S mutation of SPINK I is the most frequent genetic risk factor associated with IP. The CT gene carries several variations that could be associated with chronic pancreatitis. To avoid overestimating the pathogenetic impact of novel trypsinogen variants, a detailed clinical characterization of all patients with early onset chronic pancreatitis is mandatory.

Topics: Adult; Base Sequence; Chronic Disease; Cohort Studies; Female; Genetic Predisposition to Disease; Genetic Testing; Humans; Male; Middle Aged; Molecular Sequence Data; Mutation; Nuclear Proteins; Pancreatitis; Pedigree; Polymerase Chain Reaction; Sensitivity and Specificity; Trypsin; Trypsinogen

Mutations of the pancreatic serine protease inhibitor, Kazal type 1 (SPINK1), the cationic trypsinogen (PRSS1) and the cystic fibrosis transmembrane conductance regulator (CFTR) were reported to be genetic risk factors of chronic pancreatitis (CP). The aim of this study was to determine the role of genetic variants of the main serum antiproteinases alpha-1-antitrypsin (AAT) and alpha-2-macroglobulin (A2M) for the course of chronic pancreatitis.. 124 patients with non-alcoholic chronic pancreatitis (with PRSS1 or SPINK1 mutations or idiopathic pancreatitis) and 64 healthy controls were investigated for the AAT mutations PiS and PiZ, and the PiM determining variants R101H, V213A, E376D. In 101 subjects, the 'bait region' of A2M was sequenced. A pentanucleotide deletion in the bait region of A2M was analysed in 147 chronic pancreatitis (CP) patients and 87 controls.. The lowest prevalences of V213A and E376D were found in PRSS1 patients, whereas an increased rate of these mutations was present in the SPINK1 group, and the highest prevalence was found in patients with idiopathic pancreatitis. The prevalence of PiM variants was higher in patients with early onset CP than in late onset (P < 0.05 for E376D). The coding region of the bait region of A2M was of wild type in all investigated subjects. The A2M pentanucleotide deletion showed a homogenous distribution in patients and controls.. Our study suggests a moderating, but not predominant, role of AAT variants in the course of chronic non-alcoholic pancreatitis.

Topics: Adolescent; Adult; Aged; alpha 1-Antitrypsin; alpha-Macroglobulins; Base Sequence; Case-Control Studies; Chi-Square Distribution; Child; Child, Preschool; Chronic Disease; Cohort Studies; DNA Mutational Analysis; Female; Genetic Markers; Genetic Testing; Humans; Male; Middle Aged; Molecular Sequence Data; Mutation; Pancreatitis; Polymerase Chain Reaction; Probability; Prognosis; Sensitivity and Specificity; Serine Proteinase Inhibitors; Severity of Illness Index; Trypsin; Trypsinogen

The lysosomal cysteine protease cathepsin B is thought to play a central role in intrapancreatic trypsinogen activation and the onset of experimental pancreatitis. Recent in vitro studies have suggested that this mechanism might be of pathophysiological relevance in hereditary pancreatitis, a human inborn disorder associated with mutations in the cationic trypsinogen gene. In the present study evidence is presented that cathepsin B is abundantly present in the secretory compartment of the human exocrine pancreas, as judged by immunogold electron microscopy. Moreover, pro-cathepsin B and mature cathepsin B are both secreted together with trypsinogen and active trypsin into the pancreatic juice of patients with sporadic pancreatitis or hereditary pancreatitis. Finally, cathepsin B- catalyzed activation of recombinant human cationic trypsinogen with hereditary pancreatitis-associated mutations N29I, N29T, or R122H were characterized. In contrast to a previous report, cathepsin B-mediated activation of wild type and all three mutant trypsinogen forms was essentially identical under a wide range of experimental conditions. These observations confirm the presence of active cathepsin B in the human pancreatic secretory pathway and are consistent with the notion that cathepsin B-mediated trypsinogen activation might play a pathogenic role in human pancreatitis. On the other hand, the results clearly demonstrate that hereditary pancreatitis-associated mutations do not lead to increased or decreased trypsinogen activation by cathepsin B. Therefore, mutation-dependent alterations in cathepsin B-induced trypsinogen activation are not the cause of hereditary pancreatitis.

Topics: Blotting, Western; Cathepsin B; Cations; Dose-Response Relationship, Drug; Humans; Hydrogen-Ion Concentration; Immunohistochemistry; Mutagenesis, Site-Directed; Mutation; Pancreas; Pancreatitis; Plasmids; Recombinant Proteins; Time Factors; Trypsinogen

In the last 5 years, mutations in three genes, the cystic fibrosis transmembrane conductance regulator (CFTR) gene, the cationic trypsinogen (PRSS1) gene, and the pancreatic secretory trypsin inhibitor (PSTI) gene, have been found to be associated with chronic pancreatitis (CP). In this study, using established mutation screening methods, we systematically analysed the entire coding sequences and all exon/intron junctions of the three genes in 39 patients with idiopathic CP (ICP), with a view to evaluating the relative contribution of each gene to the aetiology of the disease. Our results demonstrate that, firstly, 'gain-of-function' mutations in the PRSS1 gene may occasionally be found in an obvious ICP subject. Secondly, presumably 'loss-of-function' mutations in the PSTI gene appear to be frequent, with a detection rate of at least 10% in ICP and, finally, abnormal CFTR alleles are common: at least 20% of patients carried one of the most common CFTR mutations, and about 10% of patients were compound heterozygotes, having at least one 'mild' allele. Thus, in total, about 30% of ICP patients carried at least one abnormal allele in one of the three genes, and this is the most conservative estimate. Moreover, a trans-heterozygous state with sequence variations in the PSTI/CFTR genes was found in three patients. However, an association between the 5T allele in intron 8 of the CFTR gene and ICP remains unproven.

Topics: Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Female; France; Genetic Predisposition to Disease; Humans; Male; Mutation; Pancreatitis; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Although chronic pancreatitis is associated with risk factors such as alcoholism, hyperparathyroidism, and hypertriglyceridaemia, little is known of the actual aetiology of the disease. It is thought that inappropriate activation of trypsinogen causes pancreatitis, and indeed in cases of hereditary pancreatitis mutations of cationic trypsinogen (PRSS1) have been described. As serine protease inhibitor Kazal type 1 (SPINK1) is a potent natural inhibitor of pancreatic trypsin activity, we hypothesised that SPINK1 mutations would be more common than expected among an unselected cohort of adult chronic pancreatitis patients.. To detect the prevalence of SPINK1 mutations in a cohort of chronic pancreatitis patients.. DNA was isolated from a cohort of 115 adult patients with chronic pancreatitis of alcoholic (n=72), hereditary (n=10), idiopathic (n=24), and miscellaneous (n=9) origin. We performed mutational analysis for two PRSS1 mutations (R122H, N29I) and four specific SPINK1 gene mutations (M1T, L14P, N34S, P55S) and compared the results with a control group of 120 healthy Dutch subjects.. In six of the 10 patients that fulfilled the criteria for hereditary pancreatitis, but in none of the control subjects, mutations in the PRSS1 gene were found. In 14 patients we detected a SPINK1 mutation. Eleven patients were heterozygous for the N34S mutation and sequencing confirmed the homozygous state of N34S in a brother and sister. Two patients carried the P55S mutation, one as a compound heterozygote with N34S. The M1T and L14P SPINK1 mutations were not found in our cohort. The N34S mutation was detected in only two of 120 controls, while the P55S, M1T, and L14P mutations were absent in the same group. Patients with the N34S allele had a later onset of disease than those with PRSS1 gene mutations but earlier onset compared with the mutation negative group.. Identification of SPINK1 mutations in 12.2% of patients with adult alcoholic and idiopathic chronic pancreatitis suggests an important role for SPINK1 as a predisposing factor in adult chronic pancreatitis.

Topics: Adult; Aged; Base Sequence; Chronic Disease; Cohort Studies; DNA Mutational Analysis; Female; Genetic Predisposition to Disease; Genotype; Humans; Male; Middle Aged; Molecular Sequence Data; Mutation; Pancreatitis; Pancreatitis, Alcoholic; Phenotype; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Endoscopic retrograde cholangiopancreatography (ERCP)-induced pancreatitis (EIP) provides an opportunity to study different pathophysiologic events early in the course of acute pancreatitis.. To investigate whether the leakage of pancreatic proenzymes (anionic trypsinogen), pancreatic protease activation (carboxypeptidase B activation peptide), cytokine response (interleukin [IL]-1 receptor antagonist, IL-6, and soluble tumor necrosis factor receptor-I) and neutrophil activation (neutrophil gelatinase-associated lipocalin and polymorphonuclear elastase) differ between patients with and without EIP. A second aim was to clarify the temporal relation between these different events.. Ninety-nine nonconsecutive patients undergoing ERCP were investigated in the study.. Fourteen of 99 patients undergoing ERCP developed mild EIP. Six hours after the investigation the concentration of anionic trypsinogen was significantly higher in patients with EIP than in patients without EIP. The day after ERCP, higher concentrations of anionic trypsinogen, carboxypeptidase B activation peptide, IL-6, and polymorphonuclear elastase were recorded in the EIP group. No significant differences in IL-1 receptor antagonist, soluble tumor necrosis factor receptor-I or neutrophil gelatinase-associated lipocalin were found between the groups in this study.. Mild EIP was accompanied by early leakage of proenzymes and later activation of trypsinogen/proteases. A significant cytokine response and neutrophil activation were recorded the day after ERCP, but further studies are needed to determine the temporal relation between these different pathophysiologic events.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; Cholangiopancreatography, Endoscopic Retrograde; Etanercept; Female; Humans; Immunoglobulin G; Interleukin 1 Receptor Antagonist Protein; Male; Middle Aged; Neutrophils; Pancreatitis; Peptides; Receptors, Tumor Necrosis Factor; Sialoglycoproteins; Trypsinogen

Cells respond to stress by upregulating the synthesis of cytoprotective heat shock proteins (HSPs) and antioxidant enzymes. The aim of this study was to compare the effects of cold (CWI) or hot water immersion (HWI) stress on three different acute pancreatitis models (cholecystokinin octapeptide (CCK), sodium taurocholate (TC), and L-arginine (Arg)). We examined the levels of pancreatic HSP60, HSP72, and antioxidants after the water immersion stress. Male Wistar rats were injected with CCK, TC, or Arg at the peak level of pancreatic HSP synthesis, as determined by Western blot analysis. HWI significantly elevated HSP72 expression and CWI significantly increased HSP60 expression in the pancreas. Water immersion stress decreased the levels of pancreatic antioxidants. CWI and-HWI pretreatment ameliorated most of the examined laboratory and morphological parameters of CCK-induced pancreatitis. CWI pretreatment decreased pancreatic edema and the serum amylase level; however, the morphological damage was more severe in TC-induced acute pancreatitis. Overall, CWI and HWI pretreatment only decreased the serum cytokine concentrations in Arg-induced pancreatitis. CWI and HWI resulted in differential induction of pancreatic HSP60 and HSP72 and the depletion of antioxidants. The findings suggest the possible roles of HSP60 and (or) HSP72 (but not that of the antioxidant enzymes) in the protection against CCK- and TC-induced acute pancreatitis. Unexpectedly, CWI pretreatment was detrimental to the morphological parameters of TC-induced pancreatitis. It was demonstrated that CWI and HWI pretreatment only influenced cytokine synthesis in Arg-induced pancreatitis.

Topics: Acute Disease; Amylases; Animals; Antioxidants; Blotting, Western; Body Weight; Chaperonin 60; Cytokines; Disease Models, Animal; Heat-Shock Proteins; HSP72 Heat-Shock Proteins; Immersion; Lipase; Male; Microscopy, Electron; Organ Size; Pancreas; Pancreatitis; Rats; Rats, Wistar; Sincalide; Stress, Physiological; Trypsinogen

Acute pancreatitis is a known complication of cardiac surgery with cardiopulmonary bypass but amylase is not a reliable marker in infants. We evaluated whether the serum concentrations of trypsinogen-2 and trypsin-2-alpha1-antitrypsin (AAT) can be used to study disturbances in pancreatic function in children and infants undergoing cardiac surgery. The study comprised 21 infants < 1 year and 25 children aged 1-16 years undergoing cardiopulmonary bypass at the Children's Hospital, Helsinki University Central Hospital. Consecutive serum samples were taken before surgery, at 12 h, 1, 2 and 3 days after surgery, and before discharge from the hospital. A moderate increase in trypsinogen-2 and trypsin-2-AAT in serum was found in more than two-thirds of the patients. On day 3, there was a 4.3-fold mean increase (CI 95% 2.8-6.5) in trypsinogen-2 and a 2.4-fold mean increase (CI 95% 1.8-3.1) in trypsin-2-AAT. In 4 patients trypsinogen-2 was elevated by more than 20-fold. One patient had clinical pancreatitis, but there were no clinical signs of pancreatitis in the other three patients. The changes in trypsinogen-2 and trypsin-2-AAT were similar in infants and children. The moderate increase in the serum concentrations of trypsinogen-2 and trypsin-2-AAT after cardiac surgery in the absence of signs of pancreatitis may be due to a subclinical pancreatic disturbance, but it could also be caused by an inflammatory response and expression of extrapancreatic trypsin. Contrary to amylase, trypsinogen-2 is expressed in the pancreas of infants.

Topics: Acute Disease; alpha 1-Antitrypsin; Amylases; Biomarkers; C-Reactive Protein; Cardiopulmonary Bypass; Child; Child, Preschool; Humans; Infant; Pancreatitis; Postoperative Complications; Trypsin; Trypsinogen

Topics: Adult; Base Sequence; Calcinosis; Chronic Disease; Diabetes Complications; DNA Mutational Analysis; Female; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Mutation; Pancreatitis; Phenotype; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen

Nontoxic heat shock protein (HSP) inducer compounds open up promising therapeutic possibilities by activating one of the natural and highly conserved defense mechanisms of the organism.. In the present experiments, we examined the effects of a HSP coinducer drug-candidate, BRX-220, on the cholecystokinin-octapeptide (CCK)-induced acute pancreatitis in rats.. Male Wistar rats weighing 240 to 270 g were divided into two groups. In group B, 20 mg/kg BRX-220 was administered orally, followed by 75 microg/kg CCK subcutaneously three times, after 1, 3, and 5 h. This whole procedure was repeated for 5 d. The animals in group slashed circleB received physiological saline orally instead of BRX-220, but otherwise the protocol was the same as in group B. The rats were exsanguinated through the abdominal aorta 12 h after the last administration of CCK. We determined the serum amylase activity, the plasma trypsinogen activation peptide concentration, the pancreatic weight/body weight ratio, the DNA and total protein contents of the pancreas, the levels of pancreatic HSP60 and HSP72, the activities of pancreatic amylase, lipase, trypsinogen, and free radical scavenger enzymes (superoxide dismutase, catalase, and glutathione peroxidase), the degree of lipid peroxidation, protein oxidation, and the reduced glutathione level. Histopathological investigation of the pancreas was also performed in all cases.. Repeated CCK treatment resulted in the typical laboratory and morphological changes of experimentally induced pancreatitis. The pancreatic levels of HSP60 and HSP72 were significantly increased in the animals treated with BRX-220. In group B, the pancreatic total protein content and the amylase and trypsinogen activities were significantly higher vs. group slashed circleB. The plasma trypsinogen activation peptide concentration, and the pancreatic lipid peroxidation, protein oxidation, and the activity of Cu/Zn-superoxide dismutase were significantly decreased in group B vs. group slashed circleB, whereas the glutathione peroxidase activity was increased. The morphological damage in group B was significantly lower than that in group slashed circleB.. The HSP coinducer BRX-220, administered for 5 d, has a protective effect against CCK-induced acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Blotting, Western; Catalase; Chaperonin 60; Glutathione; Glutathione Peroxidase; Heat-Shock Proteins; HSP72 Heat-Shock Proteins; Hydroxylamines; Lipase; Lipid Peroxidation; Male; Pancreas; Pancreatitis; Rabbits; Rats; Rats, Wistar; Sincalide; Superoxide Dismutase; Trypsinogen

Hereditary pancreatitis (HP) was defined on a clinical basis alone until the first cationic trypsinogen gene (PRSS1) mutation was discovered through the initial phase of the current Pittsburgh Midwest Multi-Center Pancreatic Study Group (MMPSG) HP study in 1996, making genetic testing available.. To evaluate the regional distribution of HP in the United States, and to compare the study's gene mutation database with the pedigree databases to determine whether family history alone predicts the likelihood of detecting mutations in the cationic trypsinogen gene.. Probands of families with HP, familial pancreatitis and idiopathic chronic pancreatitis were recruited through referrals from MMPSG collaborating centers, other physicians and self-referral of patients who had learned of the study through the World Wide Web (www.pancreas.org). Pedigrees were constructed, detailed questionnaires were completed and a blood sample was drawn for each proband and participating family members. The birthplace and current location of each patient was recorded, DNA was analyzed for known mutations and the pattern of phenotype inheritance was determined from analysis of each pedigree.. A total of 717 individuals were ascertained; 368 (51%) had clinical pancreatitis confirmed and the rest were primarily unaffected family members used for linkage studies. Forty-six clinically unaffected individuals were silent mutation carriers (11% of mutation-positive individuals). HP was most common in Minnesota, New York and the central mid-Atlantic states plus Kentucky and Ohio. One hundred and fifteen of 150 kindreds fulfilled the strict definition of an HP family, and 60 (52%) had PRSS1 mutations. Of the families with a detected mutation, 11% did not fulfill the clinical definition of an HP kindred.. The distribution of HP within the United States shows major regional differences. The etiology of HP can be identified in a small majority of HP families through genetic testing. However, family history alone is not a good predictor of finding a mutation in the cationic trypsinogen (PRSS1) gene.

Topics: Databases, Factual; Genetic Testing; Humans; Multicenter Studies as Topic; Mutation; Pancreatitis; Pedigree; Trypsinogen; United States

Three-point mutations (R117H, N211, A16V) within the cationic trypsinogen gene have been identified in patients with hereditary pancreatitis (HP). A genetic background has also been discussed for idiopathic juvenile chronic pancreatitis (IJCP), which closely mimicks the clinical pattern of HP, and alcoholic chronic pancreatitis because only a small number of heavy drinkers develop pancreatitis. This prompted us to screen 104 patients in our well-defined pancreatitis cohort for the currently known cationic trypsinogen gene mutations. The R117H mutation was detected in seven patients (six patients of two clinically classified HP families, one patient with clinically classified IJCP) and the A16V mutation in one IJCP patient. No cationic trypsinogen gene mutations were found in the remaining 96 patients with chronic and recurrent acute pancreatitis of various etiologies. Our results demonstrate the need for genetic testing to exclude HP, particularly in the presence of an atypical or unknown family history. In addition, cationic trypsinogen gene mutations are no predisposing factor in patients with chronic and recurrent acute pancreatitis of different etiologies.

Topics: Acute Disease; Adult; Child; Chronic Disease; DNA Mutational Analysis; DNA Primers; Female; Humans; Male; Middle Aged; Mutation; Pancreatitis; Pedigree; Polymerase Chain Reaction; Recurrence; Trypsin; Trypsinogen

Rapid determination of the etiology of acute pancreatitis (AP) enables institution of appropriate treatment. We evaluated the ability of trypsinogen-1, trypsinogen-2, trypsin-1-alpha(1)-antitrypsin (AAT), and trypsin-2-AAT in serum to identify the etiology of AP.. The study consisted of 67 consecutive patients with AP admitted to Helsinki University Central Hospital. Forty-two had alcohol-induced AP, 16 had biliary AP, and 9 had unexplained etiology. Serum samples were drawn within 12 h after admission. Trypsinogen-1, trypsinogen-2, trypsin-1-AAT, and trypsin-2-AAT were determined by time-resolved immunofluorometric assays. Logistic regression was used to estimate the ability of the serum analytes to discriminate between alcohol-induced and biliary AP. The validity of the tests was evaluated by ROC curve analysis.. Patients with alcohol-induced AP had higher median values of trypsin-1-AAT (P = 0.065), trypsinogen-2 (P = 0.034), and trypsin-2-AAT (P <0.001) than those with biliary AP, who had higher values of amylase (P = 0.002), lipase (P = 0.012), and alanine aminotransferase (P = 0.036). The ratios of trypsin-2-AAT to trypsinogen-1, lipase, or amylase efficiently discriminated between biliary and alcohol-induced AP (areas under ROC curves, 0.92-0.96).. Trypsinogen-2 and trypsin-2-AAT are markedly increased in AP of all etiologies, whereas trypsinogen-1 is increased preferentially in biliary AP. The trypsin-2-AAT/trypsinogen-1 ratio is a promising new marker for discrimination between biliary and alcohol-induced AP.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Alcohol Drinking; alpha 1-Antitrypsin; Biliary Tract Diseases; Female; Fluoroimmunoassay; Humans; Isoenzymes; Male; Middle Aged; Pancreatitis; ROC Curve; Trypsin; Trypsinogen

Early prediction of severity is important in the management of patients with acute pancreatitis. The presence of activation peptides and certain pancreatic proenzymes in plasma and urine has been shown to correlate with severity. This study was designed to assess the value of measuring levels of the activation peptide of carboxypeptidase B (CAPAP) and of anionic trypsinogen.. Concentrations of CAPAP and anionic trypsinogen were measured in the urine and serum in 60 patients with acute pancreatitis. Preset cut-off levels were used to analyse the accuracy of the tests. Severity was classified retrospectively according to the Atlanta classification.. Concentrations of CAPAP in urine and serum and of anionic trypsinogen in urine correlated with the severity of the pancreatitis. CAPAP in urine showed the highest accuracy. The overall accuracy was 90 per cent, with a positive predictive value of 69 per cent and a negative predictive value of 98 per cent.. In this study, measurement of CAPAP in urine was an accurate way to predict the severity of acute pancreatitis, and was superior to assay of anionic trypsinogen in urine and serum. Measurement of CAPAP in urine may be of value in the management of individual patients with pancreatitis and in the selection of patients for therapeutic trials.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Analysis of Variance; Enzyme-Linked Immunosorbent Assay; Female; Humans; Male; Middle Aged; Pancreatitis; Peptides; Radioimmunoassay; Sensitivity and Specificity; Trypsin; Trypsinogen

The aim of the study was to compare the recently introduced laboratory markers trypsinogen-2 and trypsin-2-alpha1 antitrypsin complex (trypsin-2-AAT) in serum with lipase and amylase in the diagnostic and prognostic evaluation of patients with acute pancreatitis (AP).. The analytes were measured on admission in 64 consecutive patients with AP and in 30 controls with acute abdominal disease of extrapancreatic origin. Twenty-one patients had severe and 43 mild AP. As reference methods we used serum amylase and C-reactive protein.. In subjects with AP, elevated trypsinogen-2 values (> or = 90 microg/L) were observed in 63 patients (98%), trypsin-2-AAT values (> or = 12 microg/L) in 64 patients (100%), lipase values (> or = 200 U/L) in 64 patients (100%), and amylase values (> or = 300 IU/L) in 62 patients (97%). The diagnostic accuracy of the markers was evaluated by receiver operating characteristic (ROC) analysis. On admission, trypsinogen-2, trypsin-2-AAT, lipase, and amylase differentiated patients with AP from controls with high accuracy and ROC analyses showed similar areas under the ROC curves (AUC) for trypsinogen-2 (AUC 0.960), trypsin-2-AAT (0.948), lipase (AUC 0.947), and amylase (AUC 0.930). For differentiation between severe and mild AP, trypsin-2-AAT (AUC 0.805) was slightly better than trypsinogen-2 (AUC 0.792), and they were both clearly better than lipase (AUC 0.583), C-reactive protein (AUC 0.519), or amylase (AUC 0.632) (p < 0.05).. All the markers studied showed high accuracy for differentiating between AP and extrapancreatic diseases. However, trypsinogen-2 and trypsin-2-AAT displayed the best accuracy for predicting a severe AP already at admission, which makes these markers superior for clinical purposes.

Topics: Acute Disease; alpha 1-Antitrypsin; Amylases; Case-Control Studies; Child; Female; Humans; Lipase; Middle Aged; Pancreatitis; Predictive Value of Tests; Prognosis; ROC Curve; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen

Several missense mutations, including R122H, N29I, K23R, A16V and D22G, in the cationic trypsinogen gene (PRSS1), have been associated with certain forms of hereditary pancreatitis (HP). Their occurrence in the idiopathic chronic pancreatitis (ICP) and whether novel mutations could be identified in PRSS1 remain to be further evaluated. These were addressed by the mutational screening of the entire coding sequence and the intronic/exonic boundaries of the PRSS1 gene in 221 ICP subjects, using a previously established denaturing gradient gel electrophoresis technique. Among the known PRSS1 mutations, only the R122H was detected in a single subject and the A16V in two subjects in the cohort, strengthening that HP-associated PRSS1 mutations are rare in ICP. Additional missense mutations, including P36R, E79K, G83E, K92N and V123M, were identified once separately. By analogy with the known PRSS1 mutations, predisposition to pancreatitis by some of them, particularly the V123M autolysis cleavage site mutation, is suspected. Functional analysis is expected to clarify their possible medical consequences.

Topics: Amino Acid Sequence; Chronic Disease; Cohort Studies; DNA Mutational Analysis; Humans; Molecular Sequence Data; Mutation; Mutation, Missense; Pancreatitis; Trypsin; Trypsinogen

Hyperthermia, raising the body temperature from normal to above 40 degrees C, has been shown to prevent pancreatitis in an experimental animal model of the disease, but the underlying cellular mechanisms of this protection remain unknown. We induced controlled hyperthermia in either laboratory rats and isolated pancreatic acini or, alternatively, raised the temperature of pancreatic homogenates in vitro from 37 to 41 degrees C. In vitro controlled hyperthermia of up to 41 degrees C increased the autoactivation-induced and enterokinase-induced trypsinogen activation as well as free trypsin activity. Conversely, in whole animal studies and in living acinar cells hyperthermia reduced or abolished premature intracellular trypsinogen activation in a time- and temperature-dependent manner and this protective effect was independent of either de novo protein synthesis, interference with acinar cell signal transduction, or confirmational changes in pancreatic trypsinogen. We conclude that hyperthermia, in a manner that is independent of the synthesis of pancreatic chaperone or heat shock proteins, can directly abolish the earliest initiating event involved in the onset of pancreatitis, namely the premature and intracellular activation of digestive zymogens.

Topics: Animals; Enzyme Activation; Hyperthermia, Induced; Male; Pancreas; Pancreatitis; Rats; Rats, Wistar; Trypsin; Trypsinogen

Urine trypsinogen-2 has been suggested as a marker of damage due to acute pancreatitis. Our aim was to assess the time-course and the clinical value of this test in acute pancreatitis.. A urine trypsinogen-2 dipstick test was performed on 30 patients with acute pancreatitis upon admission to the emergency room, as well as on 30 patients with non-pancreatic acute abdominal pain, and in 30 healthy subjects.. In 53.3% of the patients with acute pancreatitis the dipstick test showed abnormal urine trypsinogen-2 whereas this test gave negative results in all patients with non-pancreatic acute abdomen and in all healthy subjects. Patients with severe acute pancreatitis had a frequency of abnormal results of urine trypsinogen-2 (8/9, 88.9%; 95% CI, 51.8-99.7%) significantly higher (P = 0.031) than those with the mild disease (8/21, 38.1%; 95% CI, 18.1 -61.6%), while no significant differences were found in the urine trypsinogen-2 results between patients with biliary acute pancreatitis and those with non-biliary acute pancreatitis. Regarding the time-course of urine trypsinogen-2, there were no significant differences during the three days of the study.. The specificity of urine trypsinogen-2 in the diagnosis of acute pancreatitis is good however its sensitivity is low.

Topics: Acute Disease; Adult; Aged; Amylases; Female; Humans; Lipase; Male; Middle Aged; Pancreatitis; Prospective Studies; Sensitivity and Specificity; Trypsin; Trypsinogen

Authors report two cases of childhood chronic pancreatitis, causing severe symptoms and common bile duct stenosis with cholestasis. Both patients had to be operated on. Chronic pancreatitis with calcification led to significant common bile duct stenosis in a 13 years old girl. After ERCP a double bypass procedure was performed (Wirsungo-jejunostomy and hepatico-jejunostomy). During 42 months follow-up the patient remained pain- and symptom-free gaining 16 kilograms. In a 9 years old girl severe stenosis of the intrapancreatic common bile duct and a small duct type chronic pancreatitis with extensive fibrosis was found. Treatment was Roux-en-Y hepatico-jejunostomy. Thirty-four months after the operation she is symptom-free with normal enzyme parameters. Authors report results of genetic investigations performed on registered chronic pancreatitis children and their families in Hungary, including the two operated cases. Two of the 5 patients were hereditary type, despite negative family history. Cationic trypsinogen gene R122H (R117H) mutation were detected in both patients. Chronic non-hereditary pancreatitis is a very rare disease in childhood but may cause severe secondary conditions requiring surgery.

Topics: Adolescent; Biliary Tract Surgical Procedures; Child; Cholestasis; Chronic Disease; Common Bile Duct; Constriction, Pathologic; Female; Fibrosis; Humans; Hungary; Male; Mutation; Pancreatitis; Trypsinogen

Hereditary pancreatitis, an autosomal dominant disease is believed to be caused by mutation in the human trypsinogen gene. The role of mutations has been investigated by in vitro studies using recombinant rat and human trypsinogen (TG). In this study we compare the enzymatic properties and inhibition by human pancreatic secretory trypsin inhibitor (hPSTI) of the native, postsynthetically modified and recombinant cationic trypsin, and found these values practically identical. We also determined the autolytic stability of recombinant wild type (Hu1Asn21) and pancreatitis-associated (Hu1Ile21) trypsin. Both forms were equally stable. Similarly, we found no difference in the rate of activation of the two zymogens by human cationic and anionic trypsin. Mesotrypsin did not activate either form. The rate of autocatalytic activation of Hu1Asn21 TG and Hu1Ile21 TG was also identical at pH 8 both in the presence and absence of Ca2+. At pH 5 Hu1Ile21 TG autoactivated about twice as fast as Hu1Asn21 TG. The presence of physiological amount of hPSTI completely prevented autoactivation of both zymogens at pH 8 and at pH 5 as well. Cathepsin B readily activated both zymogens although Hu1Ile21 TG was activated about 2.5-3 times as fast as Hu1Asn21 TG. The presence of hPSTI did not prevent the activation of zymogens by cathepsin B. Our results underlie the central role of cathepsin B in the development of different forms of pancreatitis.

Topics: Amino Acid Substitution; Asparagine; Catalysis; Cathepsin B; Cloning, Molecular; Enzyme Activation; Escherichia coli; Humans; Hydrogen-Ion Concentration; Isoleucine; Pancreatitis; Recombinant Proteins; Trypsin; Trypsin Inhibitor, Kazal Pancreatic; Trypsinogen; Tumor Cells, Cultured

Hereditary, chronic pancreatitis is an autosomal dominant genetic disorder, frequently associated with two point mutations in the cationic trypsinogen gene. The mutations result in characteristic changes in the amino-acid sequence of trypsinogen: an arginine residue at position 117 is changed to histidine (Arg117-->His) or an asparagine residue at position 21 is replaced by isoleucine (Asn21-->Ile). Current opinion on the pathogenesis of hereditary pancreatitis suggests that the mutations lead to increased trypsin activity in the pancreatic tissue as a result of enhanced autoactivation of trypsinogen or decreased autocatalytic degradation (autolysis) of trypsin. To investigate the relationship between the altered properties of mutant trypsinogens and the pathomechanism of pancreatitis, wild-type and two mutant forms of recombinant human cationic trypsinogen were produced and autoactivation of trypsinogens and autolysis of trypsins were studied. The results indicate that trypsin stabilization (i.e. decreased autolysis) caused by the Arg117-->His mutation may contribute to the development of pancreatitis, however, the Asn21-->Ile mutation has no such effect. In contrast, enhanced autoactivation of mutant trypsinogens may contribute to the pathogenesis of both forms of hereditary pancreatitis. This notion is strongly supported by the clear correlation between the autoactivation rates of mutant trypsinogens and the severity of clinical symptoms.

Topics: Arginine; Asparagine; Chronic Disease; Enzyme Induction; Histidine; Humans; Isoleucine; Mutation; Pancreatitis; Trypsinogen

Mutations in the cationic trypsinogen gene have been detected in patients with hereditary pancreatitis (HP). This study investigates the prevalence of the R122H, N29I, A16V and -28delTCC mutations in the common, non-hereditary forms of chronic pancreatitis and in a HP family.. DNA was prepared from blood samples of 53 patients with chronic pancreatitis (36 alcoholic, 14 idiopathic and 3 hereditary), 20 alcoholic controls and 20 healthy, ethnically matched controls. The R122H and A16V mutations were identified by the polymerase chain reaction (PCR) and restriction enzyme digestion. A nested-PCR was used to identify the N29I mutation. The -28delTCC deletion and the C133807T polymorphism were sought by direct sequencing.. The R122H mutation was detected in 1 patient with alcoholic chronic pancreatitis and all 3 affected members of a HP family. The N29I, A16V and -28delTCC mutations were not detected in any of the study subjects. At the C133807T polymorphism, the C allele and C/C genotype were significantly increased in alcoholic chronic pancreatitis (p = 0.001 and p = 0.0004, respectively) while the T allele and CT genotype were significantly reduced (p = 0.001 and p = 0.004, respectively) compared to healthy controls.. Mutations of the cationic trypsinogen gene are rarely found in chronic pancreatitis patients of typical aetiology. Screening for these mutations should be considered in those with a family history consistent with hereditary pancreatitis but may also be appropriate in a well-defined subgroup of patients with non-hereditary chronic pancreatitis, i.e. those who have developed the disease before the age of 30.

Topics: Adult; Aged; Aged, 80 and over; Chronic Disease; DNA; Female; Humans; Male; Middle Aged; Mutation; Pancreatitis; Polymerase Chain Reaction; Trypsin; Trypsinogen

Intra-acinar cell activation of digestive enzyme zymogens including trypsinogen is generally believed to be an early and critical event in acute pancreatitis. We have found that the phosphatidylinositol 3-kinase inhibitor wortmannin can reduce the intrapancreatic activation of trypsinogen that occurs during two dissimilar experimental models of rodent acute pancreatitis, secretagogue- and duct injection-induced pancreatitis. The severity of both models was also reduced by wortmannin administration. In contrast, the NF-kappa B activation that occurs during the early stages of secretagogue-induced pancreatitis is not altered by administration of wortmannin. Ex vivo, caerulein-induced trypsinogen activation is inhibited by wortmannin and LY294002. However, the cytoskeletal changes induced by caerulein were not affected by wortmannin. Concentrations of caerulein that induced ex vivo trypsinogen activation do not significantly increase phosphatidylinositol-3,4-bisphosphate or phosphatidylinositol 3,4,5-trisphosphate levels or induce phosphorylation of Akt/PKB, suggesting that class I phosphatidylinositol 3-kinases are not involved. The concentration of wortmannin that inhibits trypsinogen activation causes a 75% decrease in phosphatidylinositol 3-phosphate, which is implicated in vesicle trafficking and fusion. We conclude that a wortmannin-inhibitable phosphatidylinositol 3-kinase is necessary for intrapancreatic activation of trypsinogen and regulating the severity of acute pancreatitis. Our observations suggest that phosphatidylinositol 3-kinase inhibition might be of benefit in preventing acute pancreatitis.

Topics: Acute Disease; Androstadienes; Animals; Cells, Cultured; Ceruletide; Chromones; Cytoskeleton; Dose-Response Relationship, Drug; Enzyme Activation; Enzyme Inhibitors; Lysosomes; Male; Mice; Morpholines; Necrosis; NF-kappa B; Pancreatitis; Phosphatidylinositol 3-Kinases; Phosphatidylinositol Phosphates; Phosphorylation; Rats; Time Factors; Trypsinogen; Wortmannin

Over the past 5 years, several gain-of-function missense mutations in the human cationic trypsinogen gene (PRSS1, OMIM 276000) have been associated with hereditary and/or sporadic pancreatitis. This study reports a new pancreatitis-associated mutation--R116C (CGT > TGT: c.346C > T)--in the gene.

Topics: Base Sequence; DNA; DNA Mutational Analysis; Humans; Mutation, Missense; Pancreatitis; Trypsin; Trypsinogen

Early identification of patients at risk of developing a severe attack of acute pancreatitis (AP) is of great importance because rapid therapeutic interventions improve outcome. At a cutoff of 50 microg/L, trypsinogen-2 measured by a rapid urinary dipstick is a sensitive and specific diagnostic test in AP. The trypsinogen-2 concentration correlates with the severity of the disease, and a test with a higher cutoff might therefore be useful for prediction of disease severity.. We increased the detection limit of the urinary trypsinogen-2 test strip (Actim Pancreatitis) from 50 microg/L to 2000 microg/L and evaluated the prognostic value of this test. The results were compared with those obtained with serum C-reactive protein and the acute physiology and chronic health evaluation II (APACHE II) score. The study population consisted of 150 consecutive patients with AP (42 with severe disease).. The sensitivity of the rapid urinary test strip (detection limit, 2000 microg/L) for prediction of severe AP, both on admission and at 24 h, was 62%; specificities were 87% and 85%, respectively, positive predictive values were 65% and 62%, and negative predictive values were 85% and 85%. C-Reactive protein had a sensitivity of only 38% on admission, but at 24 h, it was 83%; specificities were 90% and 70%, respectively, whereas positive predictive values were 59% and 52%, and NPVs were 79% and 91%, respectively. On admission the positive-likelihood ratio for the urinary trypsinogen-2 test strip was 4.8, and at 24 h it was 4.2; for C-reactive protein, the values were 3.7 and 2.7, respectively.. The urinary trypsinogen-2 dipstick is a simple and rapid method for prediction of severe acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; C-Reactive Protein; Chromatography; Female; Humans; Immunoassay; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Prognosis; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen

Nonalcoholic chronic pancreatitis is usually idiopathic and often associated with cystic fibrosis gene (CFTR) mutations. It is unknown whether pancreatitis risk correlates with having 1 or 2 CFTR mutations, abnormal epithelial ion transport, or mutations of other genes.. We tested 39 patients with idiopathic chronic pancreatitis (mean age at diagnosis, 33 years) for common mutations of CFTR and of genes encoding a trypsin inhibitor (PSTI) and trypsinogen (PRSS1). To exclude hereditary pancreatitis, we initially relied on family history and subsequently tested for PRSS1 mutations. Twenty subjects were tested for rare CFTR mutations (DNA sequencing) and 11 were tested for extrapancreatic CFTR function (clinical and physiologic evaluation).. Mutations were identified in 24 of 39 subjects. Nine patients had cystic fibrosis-causing mutations, 8 of whom also had mild-variable mutations. Eight others had only mild-variable mutations. Nine subjects had the N34S PSTI mutation and 1 had hereditary pancreatitis (R122H, PRSS1). Pancreatitis risk was increased approximately 40-fold by having 2 CFTR mutations (P < 0.0001), 20-fold by having N34S (P < 0.0001), and 900-fold by having both (P < 0.0001). Subjects with 2 CFTR mutations had abnormal nasal epithelial ion transport and clinical findings suggesting residual CFTR function between that in cystic fibrosis and in carriers. By contrast, subjects with only PSTI mutations had normal CFTR function.. CFTR-related pancreatitis risk correlates with having 2 CFTR mutations and reduced extrapancreatic CFTR function. The N34S PSTI mutation increased risk separately. Testing for pancreatitis-associated CFTR and PSTI genotypes may be useful in nonalcoholic pancreatitis.

Topics: Adolescent; Adult; Alleles; alpha-Amylases; Child; Chlorides; Cystic Fibrosis; Epithelium; Female; Gene Frequency; Genetic Predisposition to Disease; Genotype; Humans; Ion Transport; Male; Middle Aged; Mutation; Pancreatitis; Plant Proteins; Polymorphism, Genetic; Sweat; Trypsin Inhibitors; Trypsinogen

R122, the primary autolysis site of the human cationic trypsinogen (PRSS1), constitutes an important "self-destruct" or "fail-safe" defensive mechanism against premature trypsin activation within the pancreas. Disruption of this site by a missense mutation, R122H, was found to cause hereditary pancreatitis. In addition to a c.365G>A (CGC>CAC) single nucleotide substitution, a c.365 through 366GC>AT (CGC>CAT) gene conversion event in exon 3 of PRSS1 was also found to result in a R122H mutation. This imposes a serious concern on the genotyping of pancreatitis by a widely used polymerase chain reaction-restriction fragment length polymorphism assay, which could only detect the commonest c.365G>A variant.. DNA samples containing either the known c.365G>A or c.365 through 366GC>AT variant in exon 3 of PRSS1 were used as positive controls to establish a denaturing high performance liquid chromatography (DHPLC) assay.. DHPLC could readily discriminate the two known different mutational events resulting in the R122H mutation. More importantly, under the same experimental conditions, it identified a further mutational event that also occurs in the R122 primary autolysis site but results in a different amino acid substitution: c.364C>T (CGC>TGC; R122C).. A rapid, simple, and low-cost assay for detecting both the known and new mutations occuring in the R122 primary autolysis site of PRSS1 was established. In addition, the newly found R122C variant represents a likely pancreatitis-predisposing mutation.

Topics: Amino Acid Substitution; Chromatography, High Pressure Liquid; DNA; DNA Mutational Analysis; Gene Conversion; Genetic Predisposition to Disease; Humans; Mutation, Missense; Nucleic Acid Denaturation; Pancreatitis; Trypsin; Trypsinogen

Summary. The understanding of the pathogenesis of chronic pancreatitis is limited. Several theories (i. e. obstruction hypothesis) were suggested in the past but could not be confirmed by experimental data. As a formal description of the course of the disease, the necrosis-fibrosis concept seems to be very attractive. According to this theory, there is no significant difference in the pathogenesis of acute and chronic pancreatitis. A major step was the identification of mutations of the cationic trypsinogen, the secretory trypsin inhibitor (SPINK 1) and the cystic-fibrosis protein (CFTR) in some patients. Investigation of these mutations may significantly contribute to a better understanding of the pathogenesis of chronic pancreatitis.

Topics: Adult; Chronic Disease; Cystic Fibrosis Transmembrane Conductance Regulator; Fibrosis; Humans; Mutation; Necrosis; Pancreas; Pancreatic Neoplasms; Pancreatitis; Risk Factors; Trypsin Inhibitors; Trypsinogen

Hereditary pancreatitis is an autosomal dominant disease. Recently, the genetic defect has been mapped to chromosome 7q35 and consists mainly of a point mutation in exon 3 of the cationic trypsinogen gene which causes an Arg(CGC)-His(CAC) substitution at residue 117. In patients with hereditary pancreatitis the estimated cumulative risk for pancreatic carcinoma to age 70 approaches 40 %. Thus, the role of hereditary pancreatitis in the pathogenesis of pancreatic carcinoma is of interest.. DNA was extracted from peripheral blood (n = 16), fresh tumor tissue (n = 29) and formalin fixed and paraffin embedded tumor tissue (n = 5) of 50 patients with ductal adenocarcinoma of the pancreas. We specifically amplified exon 3 and the intronic flanking sequences of the cationic trypsinogen gene by nested PCR and performed restriction fragment length polymorphism analysis using the restriction enzyme Afl III. In patients with hereditary pancreatitis the G : A point mutation creates a recognition site for Afl III which is not present in unaffected individuals.. None of the 50 patients with ductal adenocarcinoma of the pancreas revealed the G : A point mutation in exon 3 of the cationic trypsinogen gene which is characteristic of hereditary pancreatitis. In addition sequencing of exon 3 did not reveal any other mutations in the DNA of patients with pancreatic adenocarcinoma.. Although hereditary pancreatitis markedly increases the risk for pancreatic cancer, it is rare and probably of little significance with respect to the pathogenesis of the majority of pancreatic adenocarcinomas.

Topics: Adenocarcinoma; Adolescent; Aged; Child; Child, Preschool; DNA; DNA Primers; DNA, Neoplasm; Exons; Humans; Infant; Nucleic Acid Amplification Techniques; Pancreatic Neoplasms; Pancreatitis; Point Mutation; Polymerase Chain Reaction; Risk Factors; Trypsinogen

We determined the clinical manifestations of hereditary pancreatitis in nearly 30 families.. The two trypsinogen mutations N29I and R122H were identified in a group of 550 patients with chronic pancreatitis of unclear origin. The following criteria were used to characterize the severity of chronic pancreatitis (one point each): calcifications, cysts, dilation of the pancreatic duct, diabetes, hospital treatment, and operation. Stages were defined as stage 0 (no points), stage 1 (one to two points), stage 2 (three to four points), and stage 3 (more than four points). Smoking and drinking habits were also recorded.. Six families with the N29I mutation (25 subjects with the mutation) and 21 families with the R122H mutation (76 subjects with the mutation) were identified. The median ages for the onset of disease were 11 years in N29I and 10 years in R122H patients. The severity of chronic pancreatitis and symptoms were similar for both mutations. About 26% (n = 26) of the 101 subjects carrying a mutation were asymptomatic, and 42% (n = 42) had mild disease (stage 1). Twenty-nine percent (n = 29) had moderate disease (stage 2), and only 4% (n = 4) had severe disease (stage 3).. Symptoms of patients with the N29I or R122H trypsinogen mutation were generally similar. The majority of subjects with trypsinogen mutations had mild disease or was asymptomatic.

Topics: Adolescent; Adult; Age Factors; Aged; Child; Child, Preschool; Chronic Disease; Health Behavior; Humans; Middle Aged; Mutation; Pancreas; Pancreatitis; Polymerase Chain Reaction; Severity of Illness Index; Trypsin; Trypsinogen

Hereditary pancreatitis is due to heterozygosity for gain-of-function mutations in the cationic trypsinogen gene which result in increased levels of active trypsin within pancreatic acinar cells and autodigestion of the pancreas. The number of disease-causing defects is generally considered to be low. To gain further insight into the molecular basis of this disorder, DNA sequence analysis of all five exons was performed in 109 unrelated patients with idiopathic chronic pancreatitis in order to determine the variability of the underlying mutations. Two German females and one German male were carriers of the most common N29I and R122H mutations (trypsinogen numbering system). In a Turkish proband, an arginine (CGT) to cysteine (TGT) substitution at amino acid position 116 was identified. Family screening demonstrated that the patient had inherited the mutation from his asymptomatic father and that he had transmitted it to both of his children, his daughter being symptomatic since the age of 3 years. In addition, a German male was found to be a heterozygote for a D100H (GAC-->CAC) amino acid replacement. Our data provide evidence for genetic heterogeneity of hereditary pancreatitis. The growing number of cationic trypsinogen mutations is expected to change current mutation screening practices for this disease.

Topics: Adult; Arginine; Chronic Disease; Cysteine; Exons; Germany; Humans; Male; Pancreatitis; Point Mutation; Polymerase Chain Reaction; Recurrence; Sequence Analysis, DNA; Trypsinogen; Turkey

We report a 25-year-old male with hemosuccus pancreaticus associated with hereditary pancreatitis. He was originally diagnosed as having familial chronic pancreatitis at the age of 12, because his brother was also diagnosed as having pancreatitis. No history of pancreatitis was found in their parents. The patient was admitted because of a growing pancreatic pseudocyst. While he had undergone conservative treatment for the pseudocyst, computed tomography incidentally revealed a pancreatic pseudoaneurysm. Endoscopic examination revealed spontaneous bleeding from the major papilla. Interventional embolization was successfully performed. An R122H mutation in the cationic trypsinogen gene was identified in this patient, his brother, and his mother, indicating that they have hereditary pancreatitis. To our knowledge, this is the first report of hemosuccus pancreaticus associated with hereditary pancreatitis. Mutational screening is useful for the diagnosis of hereditary pancreatitis, especially in patients whose diagnosis is inconclusive based on the traditional clinical criteria.

Topics: Adult; Amino Acid Substitution; Aneurysm, False; Chronic Disease; Exons; Female; Humans; Male; Mutation, Missense; Pancreas; Pancreatitis; Pedigree; Tomography, X-Ray Computed; Trypsinogen

This study was designed to evaluate the validity of a new rapid urinary trypsinogen-2 test strip (Actim Pancreatitis) for detection of acute pancreatitis in patients with acute abdominal pain.. A total of 525 consecutive patients presenting with abdominal pain at two emergency units was included prospectively and tested with the Actim Pancreatitis test strip. Urine trypsinogen-2 concentrations were also determined by a quantitative method. The diagnosis and assessment of severity of acute pancreatitis was based on raised serum and urinary amylase levels, clinical features and findings on dynamic contrast-enhanced computed tomography.. In 45 patients the diagnosis of acute pancreatitis could be established. The Actim Pancreatitis test strip result was positive in 43 of them resulting in a sensitivity of 96 per cent. Thirty-seven false-positive Actim Pancreatitis test strips were obtained in patients with non-pancreatic abdominal pain resulting in a specificity of 92 per cent. Nine patients with severe acute pancreatitis were all detected by the dipstick.. A negative Actim Pancreatitis strip result excludes acute pancreatitis with high probability. Positive results indicate the need for further evaluation, i.e. other enzyme measurements and/or radiological examinations. The test is easy and rapid to perform, unequivocal in its interpretation and can be used in healthcare units lacking laboratory facilities.

Topics: Abdominal Pain; Acute Disease; Adult; Aged; Aged, 80 and over; Biomarkers; Clinical Enzyme Tests; Female; Humans; Male; Middle Aged; Pancreatitis; Prospective Studies; Sensitivity and Specificity; Trypsin; Trypsinogen

Topics: Amino Acid Sequence; Cations; Chymotrypsin; Codon, Initiator; Genes, Dominant; Humans; Molecular Sequence Data; Mutation; Pancreatitis; Sequence Alignment; Terminology as Topic; Trypsin; Trypsinogen

Hereditary pancreatitis (HP) is an autosomal dominant disorder characterized by recurrent acute attacks of severe abdominal pain with an onset in early childhood. Many HP patients progress to complicated chronic pancreatitis and/or pancreatic cancer. Initially, a single mutation R117H in the cationic trypsinogen gene was detected in all affected members of five unrelated HP families. Further studies identified a second mutation (N21L) in two HP families without the R117H mutation. Before the association between cationic trypsinogen and HP was found, we detected a cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation (L327R) in all affected individuals of a family with HP. We therefore performed a mutational analysis for R117H and N21L in cationic trypsinogen in this and three additional unrelated families with HP. The R117H mutation was detected in all 9 affected members of three HP families and in 3 asymptomatic but at-risk relatives. However, neither the R117H nor the N21L mutation in the cationic trypsinogen were found in the HP family with the L327R alteration in CFTR. The L327R allele segregates with the disease within this HP family and was not detected on 360 unrelated Caucasian non-CF chromosomes. Although close to 800 different mutations have been detected in the CF gene of cystic fibrosis patients, L327R is a new alteration, not yet reported in connection with CF. The results of this study indicate that the CFTR gene may play a role in the etiology of minority of cases with HP and suggest that hereditary pancreatitis is genetically heterogeneous disease.

Topics: Chromosome Mapping; Cystic Fibrosis Transmembrane Conductance Regulator; Genetic Variation; Heterozygote; Humans; Mutation; Pancreatitis; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Trypsinogen

Trypsinogen and amylase content has been analysed by flow cytometry in individual pancreatic cells from rats with acute pancreatitis induced by pancreatic duct obstruction, from the earliest stages to 48 h after obstruction. Parallel morphological studies of the pancreas by electron microscopy and analysis of various parameters for the diagnosis of pancreatitis will allow research into the possible relationship between intracellular enzyme load and the severity of pancreatitis. Progressive increases in amylase activity in ascites and plasma, the volume of ascites, haematocrit, vacuolization, oedema and macrophage infiltration were observed between 1.5 h and 12 h after duct obstruction. A progressive increase in enzyme content was also observed in individual acinar cells at this stage. Interestingly, the larger increase was for trypsinogen, so that the trypsinogen/amylase ratio was significantly increased in all acinar cells by 12 h after duct obstruction. This represents a risk factor for the development of pancreatitis. Sections of pancreas taken from rats that had duct obstruction for 48 h showed massive dilatation and disorganization of the endoplasmic reticulum, focal apoptosis and necrosis. These severe alterations would affect enzyme synthesis, as reflected by the significant decrease in the intracellular enzyme load observed at this stage. However, not all acinar cells were affected equally by the damage induced by pancreatitis: R(1) cells appeared to be more sensitive than R(2) cells. In conclusion, intracellular accumulation of digestive enzymes occurs at early stages of pancreatitis, and this effect is proportionally greater for trypsinogen, a finding that could explain the degree of severity achieved in the course of pancreatitis.

Topics: Acute Disease; Amylases; Animals; Constriction; Flow Cytometry; Humans; Male; Microscopy, Electron; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

Hereditary pancreatitis has been shown to be caused by one of two mutations (R117H and N21I) of the cationic trypsinogen gene (PRSS1). Families with hereditary pancreatitis in the north of England were investigated for these mutations. The clinical features associated with each mutation were compared.. In individuals from nine families with hereditary pancreatitis, DNA was screened for the R117H and N21I mutations. All five exons of the cationic trypsinogen gene were also sequenced to search for additional mutations. Haplotype analysis was carried out to identify common ancestors. Clinical data were collected.. The R117H mutation was identified in three families and N21I in a further five. The R117H mutation was associated with a more severe phenotype than N21I in terms of mean(s.d.) age of onset of symptoms (8.4(7.2) versus 16. 5(7.1) years; P = 0.007) and requirement for surgical intervention (eight of 12 versus four of 17 patients respectively; P = 0.029). Haplotype analysis suggested that each mutation had arisen more than once.. Two mutations in the cationic trypsinogen gene cause hereditary pancreatitis in eight of nine families originating in this region. The R117H mutation is associated with a more severe form of the disease in terms of age at onset of symptoms and requirement for surgical intervention.

Topics: Age of Onset; Chronic Disease; DNA; Female; Haplotypes; Humans; Male; Mutation; Pancreatitis; Pedigree; Polymerase Chain Reaction; Trypsinogen

Topics: Animals; Ceruletide; Enteropeptidase; Pancreas; Pancreatitis; Rats; Trypsinogen

We investigated whether the timing of administration of contrast medium after onset of acute pancreatitis is critical in determining the magnitude of microcirculatory derangement.. An acute pancreatitis model in male Sprague-Dawley rats (225-275 g) was established by continuous infusion of cerulein (15 mg/kg per hour). The mean arterial pressure was monitored continuously by means of a femoral artery catheter. Diatrizoate (Hypaque-76), a water-soluble contrast medium, was delivered through a femoral vein catheter at doses corresponding to those given to humans, either 1, 2, or 3 hours after pancreatitis induction. In vivo microscopy and laser-Doppler flowmetry were used to investigate microcirculatory derangement. The water contents of the pancreas and lung, the malondialdehyde levels of the pancreas, and the trypsinogen activation peptide levels in the serum were measured at the end of the experiment (8 hours after infusion of cerulein).. Early administration of contrast medium (1 hour after pancreatitis induction) resulted in significantly greater changes in microcirculation and mean arterial pressure than did late administration (2 or 3 hours after pancreatitis induction). Rats given contrast medium 1 hour after induction also had highest pancreas and lung water contents, the highest pancreas malondialdehyde levels, and the highest serum trypsinogen activation peptide levels.. These results show that a water soluble contrast medium that is often used for computed tomographic imaging of the pancreas can adversely affect the pancreatic microcirculatory parameters, such as tissue perfusion and leukocyte sticking, and hemodynamics in a cerulein-induced model of acute pancreatitis. Early administration seems to cause more severe derangement of the pancreatic microcirculation.

Topics: Acute Disease; Animals; Blood Pressure; Ceruletide; Contrast Media; Diatrizoate; Laser-Doppler Flowmetry; Male; Malondialdehyde; Microcirculation; Microscopy; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsinogen

Mutation Asn-21 --> Ile in human cationic trypsinogen (Tg-1) has been associated with hereditary pancreatitis. Recent studies with rat anionic Tg (Tg-2) indicated that the analogous Thr-21 --> Ile mutation stabilizes the zymogen against autoactivation, whereas it has no effect on catalytic properties or autolytic stability of trypsin (Sahin-Tóth, M. (1999) J. Biol. Chem. 274, 29699-29704). In the present paper, human cationic Tg (Asn-21-Tg) and mutants Asn-21 --> Ile (Ile-21-Tg) and Asn-21 --> Thr (Thr-21-Tg) were expressed in Escherichia coli, and zymogen activation, zymogen degradation, and trypsin autolysis were studied. Enterokinase activated Asn-21-Tg approximately 2-fold better than Ile-21-Tg or Thr-21-Tg, and catalytic parameters of trypsins were comparable. At 37 degrees C, in 5 mm Ca(2+), all three trypsins were highly stable. In the absence of Ca(2+), Asn-21- and Ile-21-trypsins suffered autolysis in an indistinguishable manner, whereas Thr-21-trypsin exhibited significantly increased stability. In sharp contrast to observations with the rat proenzyme, at pH 8.0, 37 degrees C, autoactivation kinetics of Asn-21-Tg and Ile-21-Tg were identical; however, at pH 5. 0, Ile-21-Tg autoactivated at an enhanced rate relative to Asn-21-Tg. Remarkably, at both pH values, Thr-21-Tg showed markedly higher autoactivation rates than the two other zymogens. Finally, autocatalytic proteolysis of human zymogens was limited to cleavage at Arg-117, and no digestion at Lys-188 was detected. The observations indicate that zymogen stabilization by Ile-21 as observed in rat Tg-2 is not characteristic of human Tg-1. Instead, an increased propensity to autoactivation under acidic conditions might be relevant to the pathomechanism of the Asn-21 --> Ile mutation in hereditary pancreatitis. In the same context, faster autoactivation and increased trypsin stability caused by the Asn-21 --> Thr mutation in human Tg-1 might provide a rationale for the evolutionary divergence from Thr-21 found in other mammalian trypsinogens.

Topics: Animals; Asparagine; Catalysis; Cations; Enteropeptidase; Enzyme Activation; Enzyme Precursors; Enzyme Stability; Humans; Hydrogen-Ion Concentration; Hydrolysis; Mutagenesis, Site-Directed; Pancreatitis; Rats; Trypsinogen

Plasma and urine levels of trypsinogen activation peptides (TAP) reflect the severity of acute pancreatitis in experimental and clinical acute pancreatitis. In trypsin-taurocholate-induced pancreatitis in rats, the extrinsic bovine trypsin used for the induction of pancreatitis might influence on the TAP levels after induction of pancreatitis. The aim of the present study was to elucidate whether infused trypsin itself affects TAP levels in trypsin-taurocholate-induced pancreatitis. Rats were divided into three groups. In the pancreatitis group, acute pancreatitis was induced by a retrograde infusion of bovine trypsin and sodium taurocholate into the pancreatic duct. In the duct infusion group and peritoneal injection group, a mixture of bovine trypsin and trypsin inhibitor, ONO-3403, was infused into the pancreatic duct or the peritoneal cavity. Plasma and urine TAP concentration significantly increased in trypsin-taurocholate-induced pancreatitis but not in the duct infusion and peritoneal injection groups for 6 hours after the infusion of trypsin. Serum rat immunoreactive trypsin (IRT) and amylase significantly increased in the pancreatitis and duct infusion groups but not in the peritoneal injection group. Serum levels of bovine IRT in the pancreatitis group was significantly lower than those in duct infusion and peritoneal injection groups. In conclusion, an intraductal infusion of bovine trypsin itself into pancreatic duct does not influence the levels of plasma and urine TAP in trypsin-taurocholate-induced pancreatitis.

Topics: Acute Disease; Amylases; Animals; Cattle; Enzyme Activation; Injections, Intraperitoneal; Male; Pancreatic Ducts; Pancreatitis; Peptides; Rats; Rats, Wistar; Taurocholic Acid; Trypsin; Trypsinogen

Topics: Animals; Ceruletide; Enzyme Activation; Pancreas; Pancreatitis; Rats; Trypsinogen

Historically, trypsinogens/trypsins have been one of the most extensively studied enzyme models of protein structure and function. They have received renewed attention after the identification of mutations in the cationic trypsinogen gene as being associated with hereditary pancreatitis. A survey of the literature revealed five cloned cDNAs, but only three protein products of human trypsinogens, and their nomenclature has been confusing. The availability of the complete genomic sequencing of the human trypsinogen gene family made it possible to provide a systematic review of the genes, cDNAs, and protein products of human trypsinogens and to clarify some controversial issues. Further, the confusing coexistence of two systems for naming the cationic trypsinogen mutations is addressed.

Topics: Alternative Splicing; Amino Acid Sequence; Amino Acid Substitution; Chromosomes, Human, Pair 7; Chronic Disease; Cloning, Molecular; DNA, Complementary; Exons; Humans; Isoenzymes; Molecular Sequence Data; Multigene Family; Mutation; Pancreatic Juice; Pancreatitis; Pseudogenes; Sequence Alignment; Sequence Homology, Amino Acid; Trypsinogen

To evaluate serum feline trypsin-like immunoreactivity (fTLI) concentration and results of abdominal ultrasonography, CBC, and serum biochemical analyses for diagnosis of pancreatitis in cats.. Prospective study.. 28 cats with clinical signs compatible with pancreatitis.. Serum fTLI concentrations were determined, and abdominal ultrasonography, CBC, and serum biochemical analyses were performed prior to histologic evaluation of pancreatic, hepatic, and intestinal specimens. On the basis of histologic results, cats were categorized as having a normal pancreas (n = 10), pancreatic fibrosis with ongoing inflammation (9), pancreatic fibrosis without inflammation (4), and acute necrotizing pancreatitis (5). Serum fTLI concentrations and results of CBC, serum biochemical analyses, and histologic evaluation of hepatic and intestinal specimens were compared among groups.. Significant differences in serum fTLI concentrations or any hematologic or biochemical variable were not detected among the 4 groups of cats. Median serum fTLI concentrations were 51 micrograms/L (range, 18 to 200 micrograms/L) in cats with a normal pancreas, 32 micrograms/L (range, 12 to > 200 micrograms/L) in cats with pancreatic fibrosis and ongoing inflammation, 124 micrograms/L (range, 36 to > 200 micrograms/L) in cats with pancreatic fibrosis without ongoing inflammation, and 30 micrograms/L (range, 24 to 84 micrograms/L) in cats with acute necrotizing pancreatitis. We detected a high prevalence of concurrent hepatic and intestinal tract disease in cats with pancreatitis.. In cats with clinical signs of pancreatitis, serum fTLI concentration is poorly associated with histopathologic diagnosis.

Topics: Acute Disease; Animals; Cat Diseases; Cats; Chronic Disease; Female; Male; Pancreas; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Topics: Family Health; Humans; Mutation; Pancreatitis; Terminology as Topic; Trypsinogen

Mutations of the cationic trypsinogen have been described in hereditary pancreatitis. We report a new trypsinogen mutation in the activation peptide of the proenzyme in a family with chronic pancreatitis.. The coding region of the cationic trypsinogen gene was sequenced after polymerase chain reaction amplification. The following peptides homologous to the N-terminal end of cationic trypsinogen were synthesized (one-letter code, mutated amino acid underlined): wild-type peptide, APFDDDDKIVGG; pD22G, APFDDDGKIVGG; pK23R, APFDDDDRIVGG. The sequences of pD22G and pK23R correspond to the recently identified mutation K23R and to the mutation described here (D22G). To mimic trypsinogen activation, these peptides were digested with trypsin for 30 minutes at pH 5.0-8. 0, and the fragments were analyzed by high-performance liquid chromatography.. In a family with clinical evidence of hereditary chronic pancreatitis, a missense mutation of codon 22 (GAC-->GGC) of the cationic trypsinogen was found. This mutation results in a substitution of aspartic acid by glycine; therefore, the mutation was called D22G. Chromatographic analysis of tryptic digests of the peptides pD22G and pK23R showed hydrolysis rates of 22% and 75%, respectively, whereas the wild-type peptide was hydrolyzed at only 6%. The cleavage rates were reduced at lower pH, and no hydrolysis occurred without trypsin.. The activation peptides of the trypsinogen variants D22G and K23R could be released at a higher rate than in wild-type trypsinogen, resulting in increased amounts of trypsin in the pancreas, which could initiate pancreatitis.

Topics: Adult; Cations; Chronic Disease; DNA Mutational Analysis; Enzyme Activation; Family Health; Female; Gene Expression; Humans; Hydrogen-Ion Concentration; Male; Mutation, Missense; Oligopeptides; Pancreatitis; Pedigree; Trypsin; Trypsinogen

To establish whether serum pancreatic enzyme determination is useful in the identification of patients with chronic pancreatitis and in revealing the presence of exocrine pancreatic insufficiency. A total of 50 patients with chronic pancreatitis were included in the investigation: 19 were studied during a painful attack of the disease and 31 were observed during clinical remission of the disease and underwent a secretin-caerulein test; 21 of the 31 patients had severe pancreatic insufficiency. Thirty patients with non-pancreatic digestive diseases were also studied. Serum amylase, pancreatic isoamylase, lipase, trypsinogen and elastase- were determined in all patients.. Serum levels of the 5 enzymes studied were significantly higher in patients with pancreatic pain than in those studied during a clinical remission of the disease, and in those with non-pancreatic digestive diseases. In patients with chronic pancreatitis studied during clinical remission of the disease serum levels of pancreatic isoamylase and trypsinogen were significantly lower than in those patients with non-pancreatic digestive diseases. Considering only low serum concentrations of the five enzymes studied in diagnosing chronic pancreatitis, trypsinogen showed a sensitivity of 28%, specificity of 100%, a predictive value of a positive test of 100% and a predictive value of a negative test of 96.4%. In the 21 patients with severe pancreatic insufficiency, abnormally low serum concentrations of trypsinogen were found in 12 patients (57%), of lipase and elastase-1 in 6 (29%), of pancreatic isoamylase in 5 (24%), and of amylase in 3 (14%).. Serum pancreatic enzymes can not be considered a useful tool to identify patients with pancreatic insufficiency. However, of the five enzymes studied, serum trypsinogen appears to be a useful marker in the diagnostic work-up of chronic pancreatitis.

Topics: Adult; Aged; Aged, 80 and over; Amylases; Biomarkers; Chronic Disease; Female; Humans; Isoamylase; Lipase; Male; Middle Aged; Pancreatic Elastase; Pancreatitis; Prognosis; Sensitivity and Specificity; Severity of Illness Index; Trypsinogen

The N21I missense mutation in the cationic trypsinogen gene is the second most frequent mutation in hereditary pancreatitis (HP). In this article, we suggest that the N21I mutation most likely arose as a gene conversion event in which the functional anionic trypsinogen gene acted as the donor sequence. This hypothesis is supported by the unique presence of Ile at residue 21 of the anionic gene amongst the several highly homologous trypsinogen genes; a single unbroken tract of nucleotides of up to 113 bp flanking the I21 residue in the anionic trypsinogen gene; and the presence of a chi-like sequence in the 5' proximity and a palindromic sequence in the 3' vicinity of the N21I mutation. Furthermore, a multiple alignment of the partial amino acid sequence of vertebrate trypsins around residue 21 indicated that N21 and I21 may represent advantageously selected mutations of the two functional human trypsinogen genes in evolutionary history. These observations, which are complementary to the previous findings, provide further insights into the genetic mechanism and pathogenic role of the N21I mutation in HP.

Topics: Amino Acid Sequence; Animals; Base Sequence; Cattle; Dogs; Evolution, Molecular; Gene Conversion; Humans; Mice; Molecular Sequence Data; Multigene Family; Mutation, Missense; Pancreatitis; Rats; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Swine; Trypsin; Trypsinogen; Xenopus

Cationic trypsinogen and cystic fibrosis mutations have been identified in pancreatitis patients, although no study has looked for mutations in both genes in the same patient. Pancreatitis can be induced by alcohol, although not all alcoholics develop pancreatitis. We hypothesize that this phenomenon is due to a genetic predisposition in persons with alcohol-related pancreatitis. We performed sequence analysis of the cationic trypsinogen-coding region in 46 alcohol-related pancreatitis patients and 16 patients with pancreatitis due to causes other than alcohol. We also screened for 40 cystic fibrosis mutations including the 5T allele. No cationic trypsinogen mutations were identified. Cystic fibrosis mutation screening identified the DeltaF508 mutation in two Caucasian alcoholic patients (P<0.025). The cystic fibrosis mutation carrier frequency in African-American alcoholic patients was 3%, which was not significantly increased compared with the normal carrier frequency. The frequency of the 5T allele was not significantly increased compared with the normal population carrier frequency in either racial group. These results may suggest a role for the cystic fibrosis gene in alcohol-related pancreatitis but indicate that cationic trypsinogen mutations are not a common predisposing risk factor for alcohol-related pancreatitis. A multicenter study is necessary to attain sufficient numbers to come to a conclusion.

Topics: Adult; Aged; Alcohol-Related Disorders; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; DNA Mutational Analysis; Female; Genetic Counseling; Genetic Predisposition to Disease; Humans; Male; Middle Aged; Pancreatitis; Risk Factors; Trypsin; Trypsinogen

Autodigestion of the pancreas by its own prematurely activated digestive proteases is thought to be an important event in the onset of acute pancreatitis. The mechanism responsible for the intrapancreatic activation of digestive zymogens is unknown, but a recent hypothesis predicts that a redistribution of lysosomal cathepsin B (CTSB) into a zymogen-containing subcellular compartment triggers this event. To test this hypothesis, we used CTSB-deficient mice in which the ctsb gene had been deleted by targeted disruption. After induction of experimental secretagogue-induced pancreatitis, the trypsin activity in the pancreas of ctsb(-/-) animals was more than 80% lower than in ctsb(+/+) animals. Pancreatic damage as indicated by serum activities of amylase and lipase, or by the extent of acinar tissue necrosis, was 50% lower in ctsb(-/-) animals. These experiments provide the first conclusive evidence to our knowledge that cathepsin B plays a role in intrapancreatic trypsinogen activation and the onset of acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Apoptosis; Cathepsin B; Ceruletide; Disease Models, Animal; Edema; Enzyme Activation; Gene Deletion; Gene Targeting; Humans; Lipase; Mice; Mice, Knockout; Necrosis; Pancreas; Pancreatitis; Phenotype; Trypsinogen

Topics: Abdominal Pain; Acute Disease; Diagnosis, Differential; Evidence-Based Medicine; Female; Humans; Middle Aged; Pancreatitis; Sensitivity and Specificity; Trypsinogen

Activation of trypsinogen and phospholipase A(2) is an early event in pancreatic inflammation, but little is known about zymogen activation and the severity of human pancreatitis.. Using a new fluoroimmunoassay we measured trypsinogen activation peptide (TAP) and phospholipase A(2) activation peptide (PROP) in plasma and ascites in 25 patients with acute pancreatitis. TAP, PROP, Pro-PROP and pancreatic PLA(2)-I were measured in plasma for 14 days and in pancreatic necroses, ascitic fluid and pleural effusions.. All 16 patients with severe acute pancreatitis (SAP) had pancreatic necrosis, 10 developed systemic complications like sepsis, pulmonary or renal failure, 6 had infected necrosis, and 4 died. All 9 patients with mild pancreatitis (MAP) survived. Plasma TAP on admission was higher in patients with SAP than in those with MAP and increased in infected necroses. It did not correlate with systemic complications. Systemic PROP was not increased in complicated courses but was significantly higher in patients with MAP than in those with SAP on admission. Pro-PROP was higher in patients with SAP than in those with MAP but was not correlated with systemic complications. Plasma pancreatic PLA(2)-I was increased but not different in patients with SAP and those with MAP. In patients with pancreatic necrosis, TAP and PROP were highest, while in those with post-acute pancreatic abscess, only PROP and Pro-PROP were high. In patients with pleural effusion, TAP was low and PROP/ Pro-PROP were high.. Trypsinogen and PLA(2)-I activation are early events in acute pancreatitis and the activation peptides can be detected in plasma. In the pancreas, trypsinogen activation is accompanied by PLA(2)-I activation in patients with pancreatic necrosis. However, in our study, organ complications in SAP patients was not associated with increased plasma PROP.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Ascites; Enzyme Induction; Female; Humans; Male; Middle Aged; Necrosis; Oligopeptides; Pancreatitis; Phospholipases A; Pleural Effusion; Proteins; Trypsinogen

Topics: Amino Acid Substitution; Base Sequence; Chronic Disease; DNA; DNA Mutational Analysis; Genotype; Molecular Sequence Data; Mutation; Pancreatitis; Sequence Homology, Nucleic Acid; Trypsin; Trypsinogen

Hereditary pancreatitis (HP), an autosomal dominant disorder, has been associated with mutations in the cationic trypsinogen gene. Here we demonstrate that the two most frequent HP mutations, Arg117 --> His and Asn21 --> Ile, significantly enhance autoactivation of human cationic trypsinogen in vitro, in a manner that correlates with the severity of clinical symptoms in HP. In addition, mutation Arg117 --> His inhibits autocatalytic inactivation of trypsin, while mutation Asn21 --> Ile has no such effect. The findings strongly argue that increased trypsinogen activation in the pancreas is the common initiating step in both forms of HP, whereas trypsin stabilization might also contribute to HP associated with the Arg117 --> His mutation.

Topics: Catalysis; Enteropeptidase; Enzyme Activation; Humans; Mutation; Pancreatitis; Recombinant Proteins; Trypsinogen

Trypsinogen-2 and the trypsin-2-alpha1-antitrypsin complex are recently introduced new laboratory markers for acute pancreatitis. They show high sensitivity and specificity for acute pancreatitis on admission, but little is known on their time course profiles.. The serum concentrations of trypsinogen-2 and trypsin-2-alpha1-antitrypsin were monitored in 92 patients with verified acute pancreatitis. The follow-up period was 42 days in patients with severe acute pancreatitis (N = 73) and 9 days in mild disease (N = 19).. On admission the mean serum concentration of trypsinogen-2 was 2880 microg/l in severe and 920 microg/l in mild acute pancreatitis. These values were 32- and 10-fold the upper reference limit, respectively. Trypsin-2-alpha1-antitrypsin concentrations were 1250 microg/l (100-fold the upper reference limit) and 635 microg/l (52-fold), respectively. The differences were statistically significant (P = 0.026-0.001). The concentrations of trypsinogen-2 and trypsin-2-alpha1-antitrypsin decreased gradually during the follow-up period, but they remained elevated for the entire study period in patients with severe and mild disease.. The time course profile of trypsinogen-2 and trypsin-2-alpha1-antitrypsin is favorable for diagnosing acute pancreatitis. The elevation starts within hours after the onset of the disease and it is very steep. Both markers remain elevated longer than amylase and the magnitude of the elevation correlates with the severity of the disease. This is further evidence to support the use of trypsinogen-2 and trypsin-2-alpha1-antitrypsin for the evaluation of patients suspected of having acute pancreatitis.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin; Female; Humans; Male; Middle Aged; Pancreatitis; Trypsin; Trypsinogen

We recently experienced a rare case of chronic pancreatitis in a 13-year-old Japanese boy. Recently, in hereditary pancreatitis patients, some mutations have been identified in the trypsinogen gene. The purpose of this study was to investigate whether the same mutations could also be found in this patient. Polymerase chain reaction (PCR)-amplified products of his cationic and anionic trypsinogen genes were examined by direct sequence analysis. The gene analysis failed to show any mutation in any exons and their flanking intronic sequences of his trypsinogen genes. These findings indicate that the chronic calcifying pancreatitis in the present patient is "idiopathic", and thus a rare case of juvenile pancreatitis.

Topics: Adolescent; Calcinosis; Chronic Disease; DNA Mutational Analysis; Humans; Japan; Male; Mutation; Pancreatitis; Trypsinogen

Topics: Acute Disease; Chronic Disease; Humans; Mutation; Pancreatitis; Trypsinogen

Single-point mutations in the cationic trypsinogen gene have been reported in hereditary pancreatitis kindreds in the white population. The aim of the present study was to investigate whether similar gene mutations are present in Japanese hereditary pancreatitis kindreds.. All five exons of the cationic trypsinogen gene were amplified by polymerase chain reaction and sequenced in six Japanese families with hereditary pancreatitis.. Two types of single-point mutation in the cationic trypsinogen gene, which were identical with those reported in white families with hereditary pancreatitis, were observed in separate Japanese families with hereditary pancreatitis: 21Asn (AAC) to Ile (ATC) (N21I) in exon 2 and 117Arg (CGC) to His (CAC) (R117H) in exon 3. Pancreatitis occurred at more advanced ages in patients with the N21I mutation than in those with the R117H mutation. Besides normal polymorphisms in exons 4 and 5, no mutation was found in patients in the remaining four families with hereditary pancreatitis, 21 patients with sporadic chronic pancreatitis, or five normal subjects.. These results show heterogeneity, but no racial specificity, in the cationic trypsinogen gene mutations in hereditary pancreatitis kindreds. A distinctive clinical feature for each of the mutation types is suggested: adult onset for the N21I mutation and childhood onset for the R117H mutation.

Topics: Adolescent; Adult; Age of Onset; Amino Acid Sequence; Child; Child, Preschool; Chronic Disease; Exons; Female; Humans; Japan; Male; Molecular Sequence Data; Pancreatitis; Pedigree; Point Mutation; Polymerase Chain Reaction; Sequence Analysis, DNA; Trypsinogen

Topics: Acute Disease; Animals; Ceruletide; Disease Models, Animal; Enzyme Activation; Oligopeptides; Pancreatitis; Rats; Trypsinogen

Hereditary pancreatitis (HP) is a rare inherited disorder, characterised by recurrent episodes of pancreatitis often beginning in early childhood. The mode of inheritance suggests an autosomal dominant trait with incomplete penetrance. The gene, or at least one of the genes, responsible for hereditary pancreatitis has been mapped to the long arm of chromosome 7 and a missense mutation, an arginine to histidine substitution at residue 117 in the trypsinogen cationic gene (try4) has been shown to segregate with the HP phenotype. The aim of this work was to investigate the molecular basis of hereditary pancreatitis. This study was performed on 14 HP families. The five exons of the trypsinogen cationic gene were studied using a specific gene amplification assay combined with denaturing gradient gel electrophoresis (DGGE). The present paper describes three novel mutations, namely K23R and N29I and a deletion -28delTCC in the promoter region. We also found a polymorphism in exon 4, D162D. In eight of these families we found a mutation which segregates with the disease. A segregation analysis using microsatellite markers carried out on the other families suggests genetic heterogeneity in at least one of them. Our findings confirm the implication of the cationic trypsinogen gene in HP and highlight allelic diversity associated with this phenotype. We also show that the pattern of inheritance of HP is probably complex and that other genes may be involved in this genetic disease.

Topics: Cations; Exons; Female; Genetic Diseases, Inborn; Genetic Heterogeneity; Humans; Male; Mutation; Pancreatitis; Pedigree; Trypsinogen

Hereditary pancreatitis is a rare form of chronic recurrent pancreatitis. A family, in which 11 members had chronic pancreatitis, five had diabetes, and two had pancreatic cancer, was studied, and hereditary pancreatitis was diagnosed in all patients by demonstrating the mutation in exon 3 of the cationic trypsinogen gene (R117H). The clinical implications of genotypic analysis in hereditary pancreatitis are discussed.

Topics: Adolescent; Chronic Disease; Female; Humans; Mutation; Pancreatitis; Pedigree; Trypsinogen

The amount of enzymes stored in individual zymogen granules and the glycosylation of their membrane have been analysed in rats with acute pancreatitis induced by caerulein after hydrocortisone treatment. The consequences of prolonging hydrocortisone administration after pancreatitis and the use of the cholecystokinin (CCK) receptor antagonist, L-364,718, have also been evaluated.. Analysis was performed using flow cytometry.. Caerulein-induced pancreatitis in rats previously treated for 7 days with hydrocortisone (10 mg kg-1 per day) revealed alterations in enzyme storage in the pancreas. Significant increases in amylase and trypsinogen contents in zymogen granules were observed, an effect associated with a reduction in L-fucose glycoconjugates. Pancreatitis persists 7 days later if hydrocortisone treatment is prolonged. At this stage, a reduced granule fucosylation was still observed, and a significant decrease in the amount of trypsinogen stored in the granules was found. However, hydrocortisone administration led to an increase in intragranular amylase quantities up to normal values, even when L-364,718 was simultaneously administered, but it reverted to plasma as a consequence of pancreatitis. The amount of N-acetyl D-glucosamine in the zymogen granule membrane was not altered by caerulein acute pancreatitis induced under continuous hydrocortisone treatment, but it was decreased by the administration of L-364,718 over 7 days after pancreatitis induction.. The administration of hydrocortisone after the development of pancreatitis prevented recurrence of the disease. L-364,718 proved to be detrimental, not only failing to reduce the symptoms of pancreatitis but also altering the glycoproteins of zymogen granule membrane.

Topics: Acetylglucosamine; Acute Disease; Amylases; Animals; Ceruletide; Cytoplasmic Granules; Devazepide; Enzyme Precursors; Hematocrit; Hydrocortisone; Intracellular Membranes; Male; Pancreatitis; Rats; Rats, Wistar; Receptors, Cholecystokinin; Trypsinogen

In pancreatitis, a key role has been attributed to the inappropriate conversion of trypsinogen to trypsin. Recently, two mutations of the cationic trypsinogen gene were found in families with hereditary pancreatitis. This study was conducted to determine the spectrum and frequency of cationic trypsinogen mutations in unrelated patients with idiopathic or hereditary chronic pancreatitis (CP).. DNA samples from 44 unrelated children and adolescents with CP (30 patients with idiopathic CP and 14 with hereditary CP) and from 56 family members were investigated. The cationic trypsinogen gene was screened for mutations by single-strand conformation polymorphism analysis and DNA sequencing.. A mutation in the cationic trypsinogen gene was detected in 5 patients: in 2 patients with a family history of CP and in 3 patients with idiopathic CP. In 1 patient the formerly described R122H mutation was detected. In 4 patients a hitherto unknown mutation was found at the signal peptide cleavage site leading to an alanine to valine exchange in codon 16. The mutations were inherited in all cases. In 95 unrelated control individuals the A16V mutation was not found.. Heterozygosity for the A16V mutation is strongly associated with CP. These results indicate that a significant percentage of patients with idiopathic CP may have a genetic basis for their disorder; therefore, genetic testing should be included in the diagnostic evaluation of these patients.

Topics: Adolescent; Amino Acid Sequence; Cations; Child; Child, Preschool; Chronic Disease; Female; Humans; Infant; Male; Molecular Sequence Data; Mutation; Pancreatitis; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Protein Sorting Signals; Trypsinogen

Nine out of 48 (19%) patients referred to a pancreatic clinic with a presumed diagnosis of idiopathic chronic pancreatitis have been shown to have mutations in the cationic trypsinogen gene (PRSSI), consistent with a previously unsuspected diagnosis of hereditary pancreatitis.

Topics: Adult; Chromosome Aberrations; Chromosome Disorders; Chromosome Mapping; Chronic Disease; Diagnosis, Differential; DNA Mutational Analysis; Humans; Pancreatitis; Pancreatitis, Alcoholic; Polymerase Chain Reaction; Risk Factors; Trypsinogen

Topics: Acute Disease; Cathepsin B; Cysteine Proteinase Inhibitors; Dipeptides; Enzyme Activation; Humans; Leucine; Pancreatitis; Piperazines; Protease Inhibitors; Trypsinogen

Acute pancreatitis (AP) results in elevated concentrations of trypsinogen (T) isoenzymes in serum. Immunoreactive anionic trypsinogen in urin (irAT/u) is elevated in AP, and has recently been proposed as a rapid diagnostic instrument and severity predictor. These results have not been confirmed by other groups, and irAT/u has not been further characterized. The concentration of immunoreactive cationic trypsinogen in urine (irCT/u) and the serum irAT/irCT ratio in AP have not been extensively examined.. Levels of irAT and irCT were studied in urine and serum from 50 AP patients and in urine from 41 non-AP patients. Severity was assessed according to the Atlanta classification. irAT/u was characterized by gel filtration.. Gel filtration revealed only AT in the urine. Highly significant differences in irAT/u were seen between AP/non-AP (p < 0.0001) and mild/severe disease (p = 0.0012). The irAT/irCT ratio in serum changed from normal 0.8 to 1.3 in AP.. IrAT and only traces of irCT were found in the urine in AP. IrAT/u was higher in AP than in other acute abdominal disorders (non-AP) and also higher in severe than in mild AP. IrAT in serum (irAT/s) increased proportionally more than irCT/s in AP, but did not discriminate mild from severe forms. High levels of irAT/u in some non-AP cases and a wide range in AP cases make the clinical value of the test questionable.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; Digestive System Diseases; Enzyme-Linked Immunosorbent Assay; Female; Humans; Male; Middle Aged; Pancreatitis; Sensitivity and Specificity; Trypsin; Trypsinogen

Intracellular activation of trypsinogen is currently believed to initiate pancreatitis. Factors responsible for the progression of mild to necrotizing pancreatitis are poorly understood. This study evaluated the significance of interstitial protease release and activation in this process.. In rats with cerulein-induced pancreatitis, concentrations of trypsinogen and its activation peptide TAP were measured in lymph and blood, and pancreatic injury was determined. Activation of extracellular trypsinogen was induced by intravenous infusion of enterokinase, which does not enter the acinar cell. Gabexate mesilate (acinar cell permeable) or soybean trypsin inhibitor (acinar cell nonpermeable) was administered to distinguish the effects of intracellular or extracellular protease activation.. In cerulein pancreatitis, trypsinogen levels increased prominently and were highest in lymph and portal vein blood, whereas TAP increments were modest. Combined cerulein/enterokinase infusions resulted in marked TAP increases in lymph and blood and in severe necrohemorrhagic pancreatitis. Gabexate mesilate as well as soybean trypsin inhibitor significantly decreased TAP levels in both lymph and blood and reduced pancreatic injury, with no significant differences between groups.. In secretagogue-induced pancreatitis, large amounts of trypsinogen are present in the interstitium and drain via the portal and lymphatic circulation. Activation of this extracellular trypsinogen induces hemorrhagic necrosis in a setting of mild edematous pancreatitis. This phenomenon may be the central event in the progression to fulminant necrotizing pancreatitis.

Topics: Animals; Ceruletide; Enteropeptidase; Enzyme-Linked Immunosorbent Assay; Male; Oligopeptides; Pancreatitis; Pancreatitis, Acute Necrotizing; Rats; Rats, Sprague-Dawley; Trypsinogen

Topics: Humans; Mutation; Pancreatitis; Trypsinogen

The pancreatitis-associated protein (PAP) was investigated in patients with hereditary and chronic alcoholic pancreatitis. Blood levels of pancreatic enzymes and PAP were measured in nine families with hereditary pancreatitis; in three of them, the mutation N21I, and in six, the R117H variant of the cationic trypsinogen were present. In all family members, similar to controls, only normal values of the PAP were found. There was no evidence for polymorphism of the PAP gene in patients with hereditary or alcoholic pancreatitis. Immunohistochemically PAP was detected in the apical parts of the acinar cells but not in ducts, interstitial tissue, islets, or blood vessels. Intensity of PAP labeling was directly related to the deterioration of the acinar units, and its concentration was inversely related to chymotrypsinogen immunoreactivity in the same tissue. Similar immunohistochemical findings were present in chronic alcoholic and hereditary pancreatitis. We conclude that there is a lack of PAP polymorphism in hereditary and alcoholic pancreatitis and that expression of the PAP in both groups of patients is related to the degree of cellular damage of the pancreas.

Topics: Acute-Phase Proteins; Amino Acid Substitution; Amylases; Antigens, Neoplasm; Biomarkers, Tumor; C-Reactive Protein; Child; Chymotrypsinogen; DNA Mutational Analysis; Humans; Immunohistochemistry; Inflammation; Lectins, C-Type; Lipase; Male; Mutation; Pancreatitis; Pancreatitis-Associated Proteins; Pancreatitis, Alcoholic; Polymerase Chain Reaction; Trypsinogen

Mutations Arg(117) --> His and Asn(21) --> Ile in human trypsinogen-I have been recently associated with hereditary pancreatitis (HP). The Arg(117) --> His substitution is believed to cause pancreatitis by stabilizing trypsin against autolytic degradation, while the mechanism of action of Asn(21) --> Ile has been unknown. In an effort to understand the effect(s) of this mutation, Thr(21) in the highly homologous rat trypsinogen-II was replaced with Asn or Ile, and the recombinant zymogens and their active trypsin forms were studied. Kinetic parameters of all three trypsins were comparable, and the active enzymes suffered autolysis at similar rates, indicating that neither catalytic properties nor proteolytic stability of trypsin are influenced by mutations at position 21. When incubated at pH 8.0, 37 degrees C, pure zymogens underwent autoactivation with concomitant trypsinolytic degradation in a Ca(2+)-dependent fashion. Thus, in the presence of 5 mM Ca(2+), autoactivation and digestion of the zymogens after Arg(117) and Lys(188) were observed, while in the presence of 1 mM EDTA autoactivation and cleavage at Lys(188) were reduced, and zymogenolysis at the Arg(117) site was enhanced. Overall rates of zymogen degradation in [Asn(21)]- and [Ile(21)]trypsinogens were higher in Ca(2+) than in EDTA, while [Thr(21)]trypsinogen demonstrated inverse characteristics. Remarkably, both in the presence and absence of Ca(2+), [Ile(21)]trypsinogen exhibited significantly higher stability against autoactivation and proteolysis than zymogens with Asn(21) or Thr(21). The observations suggest that autocatalytic trypsinogen degradation may be an important defense mechanism against excessive trypsin generation in the pancreas, and trypsinogen stabilization by the Asn(21) --> Ile mutation plays a role in the pathogenesis of HP.

Topics: Animals; Asparagine; Calcium; Catalysis; Enteropeptidase; Enzyme Activation; Humans; Isoleucine; Pancreatitis; Point Mutation; Rats; Recombinant Proteins; Trypsinogen

The aim of this study was to evaluate pancreatic function in total parenteral nutrition (TPN)-dependent children with permanent intestinal failure by measuring immunoreactive trypsinogen (IRT) levels. Between 1992 and 1996, 105 pediatric patients with permanent intestinal failure were referred to the Children's Hospital of Pittsburgh for small intestinal transplant evaluation. Serum samples were available from 55 of them. Ten suffered from intestinal pseudo-obstruction or microvillus inclusion disease, while 45 had short bowel syndrome (SBS). IRT levels were significantly higher (p < 0.001) in SBS patients (89.4 +/- 9.2 ng mL) compared to controls (43.4 +/- 5.6 ng/ nL) without liver, gastrointestinal, or kidney disease. IRT levels did not correlate with liver injury, length of bowel, or the cause of SBS. Five of 20 patients who underwent intestinal transplantation developed pancreatitis during a median post-operative follow up 15.4 months later. IRT levels failed to predict who would develop pancreatitis post-transplant. The data suggest that elevated plasma IRT levels are common among children with intestinal failure, but fail to identify patients at risk for pancreatitis post-transplant.

Topics: Adolescent; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Humans; Infant; Intestinal Diseases; Intestinal Pseudo-Obstruction; Intestines; Male; Pancreatitis; Parenteral Nutrition, Total; Risk Factors; Short Bowel Syndrome; Transplantation, Homologous; Trypsinogen

Topics: Child; Chronic Disease; Humans; Molecular Biology; Mutation; Pancreatitis; Trypsinogen

Mutations Arg117-->His and Asn21-->Ile of the human cationic trypsinogen have been recently identified in patients affected by hereditary pancreatitis (HP). The Arg117-->His substitution is believed to cause pancreatitis by eliminating an essential autolytic cleavage site in trypsin, thereby rendering the protease resistant to inactivation through autolysis. Here we demonstrate that the Arg117-->His mutation also significantly inhibits autocatalytic trypsinogen breakdown under Ca(2+)-free conditions and stabilizes the zymogen form of rat trypsin. Taken together with recent findings demonstrating that the Asn21-->Ile mutation stabilizes rat trypsinogen against autoactivation and consequent autocatalytic degradation, the observations suggest a unifying molecular pathomechanism for HP in which zymogen stabilization plays a central role.

Topics: Animals; Arginine; Binding Sites; Electrophoresis, Polyacrylamide Gel; Enteropeptidase; Enzyme Activation; Enzyme Stability; Humans; Mutagenesis, Site-Directed; Mutation; Pancreatitis; Rats; Trypsin; Trypsinogen

Hereditary pancreatitis is associated with at least 2 mutations in the cationic trypsinogen gene. The purpose of the present study is to test the utility of T4 endonuclease VII for the detection of cationic trypsinogen R117H mutations. In addition, the possibility of screening for R117H, N21I, and A8V mutations in a single 2.2-kb polymerase chain reaction (PCR) product using T4 endonuclease VII was investigated.. Twenty-nine DNAs from control patients and patients with known cationic trypsinogen R117H, A8V, or N21I mutations were selected from the ongoing hereditary pancreatitis study of the Midwest Multicenter Pancreatic Study Group. The samples were coded and randomized, and a 911-bp sequence containing exon 3 or a 2,212- bp sequence containing exons 2 and 3 were amplified by PCR using fluorescent- labeled primers. The PCR products were digested with T4 endonuclease VII and screened for mutations on an automated DNA sequencer.. In all cases with a mutation, a cleavage fragment on the direct and/or complementary DNA strand could easily be visualized, and its approximate size correlated with the predicted location of the known mutations within the PCR product. When the code for affected status was broken, there was 100% correlation between previous DNA sequence or restriction fragment length polymorphism findings and the T4 endonuclease VII digestion results for all 29 DNAs.. T4 endonuclease VII accurately identified the known cationic trypsinogen gene mutations in exons 2 and 3. Enzymatic mutation detection appears to be an accurate and useful method for screening individuals for known trypsinogen gene mutations and may be useful in identifying previously unidentified mutations within large regions of interest.

Topics: Acute Disease; Amino Acid Substitution; Child; Chromosomes, Human, Pair 7; DNA; DNA Mutational Analysis; Endodeoxyribonucleases; Exons; Genes, Dominant; Genetic Predisposition to Disease; Humans; Isoenzymes; Pancreatitis; Point Mutation; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Recurrence; Trypsinogen

Pancreatic proteases are secreted in acute pancreatitis, but their contribution to associated lung injury is unclear. Applying models of mild edematous (intravenous caerulein) and severe necrotizing (intraductal glycodeoxycholic acid) pancreatitis in rats, we showed that both trypsinogen and trypsin concentrations in peripheral blood, as well as lung injury, correlate with the severity of the disease. To isolate the potential contribution of proteases to lung injury, trypsin or trypsinogen was injected into healthy rats or trypsinogen secreted in caerulein pancreatitis was activated by intravenous enterokinase. Pulmonary injury induced by protease infusions was dose dependent and was ameliorated by neutrophil depletion. Trypsinogen activation worsened lung injury in mild pancreatitis. In vitro incubation of leukocytes with trypsinogen showed that stimulated leukocytes can convert trypsinogen to trypsin. In conclusion, this study demonstrates that the occurrence and severity of pancreatitis-associated lung injury (PALI) corresponds to the levels of circulating trypsinogen and its activation to trypsin. Neutrophils are involved in both protease activation and development of pulmonary injury.

Topics: Acute Disease; Animals; Carcinogens; Ceruletide; Detergents; Endopeptidases; Enteropeptidase; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Glycodeoxycholic Acid; Leukocytes; Lung; Lung Diseases; Male; Oligopeptides; Pancreas; Pancreatitis; Peroxidase; Rats; Rats, Sprague-Dawley; Tetradecanoylphorbol Acetate; Trypsin; Trypsinogen

Little is known about the changes in pancreatic enzyme storage in acute pancreatitis. We have performed flow cytometric studies of zymogen granules from rats with acute pancreatitis induced by hyperstimulation with caerulein. A comparison was made with rats treated with hydrocortisone (10 mg/kg/day) over 7 days before inducing pancreatitis in order to find out whether the amount of enzymes stored in the pancreas plays a key role in the development of pancreatitis. The potentially therapeutic effect of L-364,718 (0.1 mg/kg/day, for 7 days), a CCK receptor antagonist, was assayed in the rats with caerulein-induced pancreatitis which had previously received the hydrocortisone treatment. A significant increase in the intragranular enzyme content was observed 5 h after hyperstimulation with caerulein. The highest values were reached in the rats previously treated with hydrocortisone. The greatest pancreatic enzyme load was parallel to the highest values in plasma amylase, edema and haematocrit observed. Acute pancreatitis was reversed seven days later. At this stage smaller granules appeared in the pancreas whose enzyme content was similar to that of controls when no treatment was applied after pancreatitis. In contrast, L-364,718 administration prevented the favourable evolution of pancreatitis since the antagonism exerted on CCK receptors induced a blockade of secretion of the large amounts of enzymes stored in the pancreas. Moreover, the enzyme content in zymogen granules was below normal values since the stimulatory CCK action on enzyme synthesis can be inhibited by L-364,718. Our results suggest that the efficiency of CCK antagonists, as potential therapy, would also depend on the load of enzymes in the pancreas when acute pancreatitis is produced.

Topics: Acute Disease; Amylases; Animals; Ceruletide; Cytoplasmic Granules; Devazepide; Enzyme Precursors; Hormone Antagonists; Hydrocortisone; Male; Pancreatitis; Rats; Rats, Wistar; Receptor, Cholecystokinin A; Receptors, Cholecystokinin; Trypsinogen

Topics: Alanine; Amino Acid Substitution; Chronic Disease; Humans; Mutation; Pancreatitis; Trypsinogen; Valine

Topics: Alanine; Amino Acid Sequence; Amino Acid Substitution; Biological Transport; Chronic Disease; Genetic Testing; Humans; Molecular Sequence Data; Mutation; Pancreatitis; Protein Sorting Signals; Sequence Homology, Amino Acid; Trypsinogen; Valine

Topics: Chronic Disease; Humans; Mutation; Pancreatitis; Trypsinogen

Topics: Acute Disease; Amylases; Biomarkers; Humans; Pancreatitis; Reagent Kits, Diagnostic; ROC Curve; Trypsin; Trypsinogen

To investigate the debated role of intracellular trypsinogen activation and its relation to lysosomal enzyme redistribution in the pathogenesis of acute pancreatitis, rats were infused with the cholecystokinin analog caerulein at 5 micrograms.kg-1.h-1 for intervals up to 3 h, and the changes were contrasted with those in animals receiving saline or 0.25 microgram.kg-1.h-1 caerulein. Saline or 0.25 microgram.kg-1.h-1 caerulein did not induce significant changes. In contrast, 5 micrograms.kg-1.h-1 caerulein caused significant hyperamylasemia and pancreatic edema within 30 min. Pancreatic content of trypsinogen activation peptide (TAP) increased continuously (significant within 15 min). TAP generation was predominantly located in the zymogen fraction during the first hour but expanded to other intracellular compartments thereafter. Cathepsin B activity in the zymogen compartment increased continuously throughout the experiments and correlated significantly with TAP generation in the same compartment. Total trypsinogen content increased to 143% with marked interstitial trypsinogen accumulation after 3 h. Supramaximal caerulein stimulation causes trypsinogen activation by 15 min that originates in the zymogen compartment and is associated with increasing cathepsin B activity in this subcellular compartment. However, a much larger pool of trypsinogen survives and accumulates in the extracellular space and may become critical in the evolution of necrotizing pancreatitis.

Topics: Acute Disease; Amylases; Animals; Cathepsin B; Cell Fractionation; Ceruletide; Edema; Enzyme Activation; Kinetics; Male; Oligopeptides; Pancreatitis; Rats; Rats, Sprague-Dawley; Subcellular Fractions; Trypsinogen

Topics: Acute Disease; Biomarkers; C-Reactive Protein; Carboxypeptidase B; Carboxypeptidases; Clinical Enzyme Tests; Enzyme Activation; Humans; Oligopeptides; Pancreatitis; Peptides; Sensitivity and Specificity; Trypsin; Trypsinogen

Hereditary pancreatitis (HP) is the most common form of chronic relapsing pancreatitis in childhood, and may account for approximately 25% of adult cases with chronic idiopathic pancreatitis. Recently, an arginine-histidine (R117H) mutation within the cationic trypsinogen gene was found in 5/5 families studied with HP. In this study we report on the results of linkage and direct mutational analysis for the common R117H mutation examined in 8 nonrelated families with hereditary pancreatitis. Two-point linkage analysis with the 7q35 marker D7S676, done initially in 4 families, yielded lod scores that were positive in 2, negative in one, and weakly positive in one. Direct mutational analysis of exon 3 of the cationic trypsinogen gene in 6 families showed that all symptomatic individuals tested were heterozygous for the R117H mutation. Also, several asymptomatic but at-risk relatives were found to be heterozygous for this mutation. Affected individuals in the remaining 2 families did not have the mutation. Radiation hybrid mapping using the Genebridge 4 panel assigned the trypsinogen gene to chromosome region 7q35, 2.9 cR distal to ETS WI-9353 and 3.8 cR proximal the dinucleotide repeat marker D7S676. The negative linkage and absence of the trypsinogen mutation in 2/8 families suggest locus heterogeneity in HP. Analysis of the R117H mutation is useful in identifying presymptomatic "at-risk" relatives and in genetic counseling. Also, it can be useful in identifying children and adults with isolated chronic idiopathic pancreatitis.

Topics: Chromosome Mapping; DNA Mutational Analysis; Female; Genetic Linkage; Genetic Markers; Humans; Hybrid Cells; Karyotyping; Male; Pancreatitis; Pedigree; Polymerase Chain Reaction; Trypsinogen

A 71-year-old woman was admitted with the suspected diagnosis of pancreatic carcinoma. As a child she had had repeated attacks of abdominal pain of undetermined cause. When aged 48 years she had developed diabetes mellitus. Her now 42-year-old daughter had from the age of 9 years suffered from repeated attacks of acute pancreatitis that had finally led to chronic pancreatitis. The patient's 15-year-old grandchild was having recurrent bouts of abdominal pain.. Imaging procedures revealed calcifications in the pancreas and an infiltrating space-occupying lesion, about 3 cm in diameter, in the head of the pancreas with lymph node and liver metastases. Cytological analysis of material aspirated from the space-occupying mass showed typical findings of ductal pancreatic carcinoma. FURTHER TESTS, TREATMENT AND COURSE: At first the patient's course was not typical for a genetically-determined disease, but the family history raised the suspicion of hereditary pancreatitis. A genetic test (Afl-III-RFLP test) demonstrated the mutation Arg 117 His in the cationic trypsinogen gene in all diseased or symptomatic family members. The patient died of the complications of the pancreatic cancer.. Genetic tests are valuable in the diagnosis of hereditary pancreatitis, because the increased cancer risk can be met by frequent examinations in affected family members.

Topics: Abdominal Pain; Acute Disease; Adolescent; Adult; Aged; Calcinosis; Chronic Disease; Family; Fatal Outcome; Female; Humans; Pancreas; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Point Mutation; Polymorphism, Restriction Fragment Length; Recurrence; Risk Factors; Tomography, X-Ray Computed; Trypsinogen; Ultrasonography

Topics: Acute Disease; alpha 1-Antitrypsin; Cholangiopancreatography, Endoscopic Retrograde; Humans; Pancreatitis; Sensitivity and Specificity; Severity of Illness Index; Trypsin; Trypsinogen

Topics: Acute Disease; Humans; Pancreatic Function Tests; Pancreatitis; Sensitivity and Specificity; Trypsin; Trypsinogen

1. After monitoring the changes associated with necrotizing acute pancreatitis in rats from early stages to 24 h after infusion of 5% sodium taurocholate in the choledocus, we characterized by flow cytometry the zymogen granules that still remained in the pancreas 18 h after sodium taurocholate infusion in order to explore whether alterations in the enzyme content and/or in the composition of the granule membrane could be related to the intracellular mechanisms involved in the development of necrotizing acute pancreatitis. 2. Significant increases in the haematocrit, plasma and peritoneal exudate amylase levels and oedema were observed from the third hour after 5% sodium taurocholate infusion onwards. Additionally, cell alterations such as hypergranulation, dilatation of the endoplasmic reticulum and autophagic vacuoles were found 3 and 6 h after infusion. DNA decrease, degranulation and necrosis were observed from 12 h after sodium taurocholate infusion onwards. 3. Flow cytometric measurements of zymogen granules isolated from rat pancreas 18 h after 5% sodium taurocholate infusion revealed a significant decrease in their internal complexity without major changes in their size. Double staining of granules with Tetragonolobus purpureus lectin, which specifically binds L-fucose and specific anti-trypsinogen or anti-amylase antisera, showed that rats with induced pancreatitis have decreased amounts of L-fucose in the membrane glycoconjugates and lower enzyme content (70% and 30% less for trypsinogen and amylase respectively). 4. A decrease in L-fucose in the membrane together with membrane abnormalities observed by electron microscopy in zymogen granules isolated 18 h after sodium taurocholate infusion indicate an altered synthesis of new granules or lysis of preformed zymogen granules which would favour differential loss of granular enzymes, mainly trypsinogen, which in turn could increase the severity of disease.

Topics: Acute Disease; Amylases; Animals; Cholagogues and Choleretics; Cytoplasmic Granules; Enzyme Precursors; Fucose; Hematocrit; Male; Microscopy, Electron; Pancreas; Pancreatitis; Rats; Rats, Wistar; Taurocholic Acid; Trypsinogen

Hereditary pancreatitis (OMIM 167800) is thought to be associated with a mutation of the exon 3 of cationic trypsinogen (Nature Genet (1996): 14:141-145). This paper reports sequence data of two independent families suffering from this disease. PCR amplificates from leukocyte or buccal swab DNA showed no mutation of exon 3 of cationic trypsinogen. Instead, in exon 2, an A-to-T tranversion was found that led to the substitution of Asn by Ile in the sixth amino acid of the active trypsin. In exons 4 and 5, silent mutations were found. In the other expressed trypsinogens, several homozygous alterations not associated to hereditary pancreatitis were identified. As a model of pathogenesis, we hypothesize that mutation of trypsinogen in exon 2 could lead to premature cleavage of the activation peptide of trypsinogen or to altered intracellular transport.

Topics: Base Sequence; Cations; DNA Primers; Female; Humans; Male; Mutation; Pancreatitis; Pedigree; Polymerase Chain Reaction; Trypsinogen

Although it is widely accepted that trypsinogen activation is an initiating event in the development of acute pancreatitis, its location inside the pancreas is not known. In our studies, acute edematous pancreatitis was induced in rats by one or two intraperitoneal injections of 50 microg cerulein/kg body weight. The pancreas was removed for examination 1 or 2 h after the first and the second cerulein injection, respectively. The cleavage product of trypsinogen activation, trypsinogen activation peptide, was specifically labeled on pancreatic tissue sections by a corresponding antibody, the signal enhanced by a biotin-avidin conjugate, and the site then visualized by coupled peroxidase activity on diaminobenzidine. The sections were examined by light microscopy. Trypsinogen activation peptide, reflecting activation of the pancreatic digestive enzyme trypsinogen, was detected inside pancreatic acinar cells in this animal model of acute pancreatitis. As early as 1 h after the first injection of cerulein, protease activation was seen within the apical pole of acinar cells. Protease activation was increased 2 h after the latter of two injections of cerulein and more evenly distributed within the cells. For the first time morphologic evidence confirms that the activation originates within the acinar cell, rather than from the interstitium or the duct lumen. The location of this activation at the apical site of the acinar cell indicates its origin from subcellular compartments involving the late steps in the secretory pathway.

Topics: Animals; Ceruletide; Disease Models, Animal; Enzyme Activation; Female; Immunoenzyme Techniques; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsin; Trypsinogen

Supramaximal stimulation of the pancreas with the CCK analog caerulein causes acute edematous pancreatitis. In this model, active trypsin can be detected in the pancreas shortly after the start of supramaximal stimulation. Incubation of pancreatic acini in vitro with a supramaximally stimulating caerulein concentration also results in rapid activation of trypsinogen. In the current study, we have used the techniques of subcellular fractionation and both light and electron microscopy immunolocalization to identify the site of trypsinogen activation and the subsequent fate of trypsin during caerulein-induced pancreatitis. We report that trypsin activity and trypsinogen-activation peptide (TAP), which is released on activation of trypsinogen, are first detectable in a heavy subcellular fraction. This fraction is enriched in digestive enzyme zymogens and lysosomal hydrolases. Subsequent to trypsinogen activation, both trypsin activity and TAP move to a soluble compartment. Immunolocalization studies indicate that trypsinogen activation occurs in cytoplasmic vacuoles that contain the lysosomal hydrolase cathepsin B. These observations suggest that, during the early stages of pancreatitis, trypsinogen is activated in subcellular organelles containing colocalized digestive enzyme zymogens and lysosomal hydrolases and that, subsequent to its activation, trypsin is released into the cytosol.

Topics: Acute Disease; Animals; Cathepsin B; Cell Fractionation; Ceruletide; Edema; Enzyme Activation; Immunohistochemistry; Kinetics; Lysosomes; Male; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Wistar; Subcellular Fractions; Time Factors; Trypsin; Trypsinogen; Vacuoles

Hereditary pancreatitis is an autosomal dominant disorder with incomplete penetrance. It is characterised by recurring episodes of severe abdominal pain and often presents in childhood. Recently, a mutation in the cationic trypsinogen gene was identified in this disease. Previously, only one mutation at residue 117 of the trypsinogen gene has been found in the five separate hereditary pancreatitis families, four from the USA and one from Italy. Alteration of the Arg117 site is believed to disrupt a fail-safe mechanism for the inactivation of trypsin, leading to autodigestion of the pancreas under certain conditions. Molecular analysis of the trypsinogen gene was carried out on a hereditary pancreatitis family from the UK. The same G to A mutation at residue 117 was identified in this family, suggesting that this is a common mutation in hereditary pancreatitis.

Topics: Chronic Disease; Female; Humans; Male; Mutation; Pancreatitis; Pedigree; Polymerase Chain Reaction; Recurrence; Trypsinogen

Topics: Acute Disease; Animals; Chronic Disease; Dogs; Humans; Pancreas; Pancreatitis; Point Mutation; Trypsin; Trypsinogen

Topics: Acute Disease; Adolescent; Biomarkers; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Humans; Infant; Male; Pancreas; Pancreatitis; Patient Selection; Reagent Kits, Diagnostic; Reference Values; Retrospective Studies; Short Bowel Syndrome; Trypsinogen

The etiology of nonalcoholic chronic pancreatitis, occurring in tropical regions, is unknown. Although environmental factors may play a role in its pathogenesis, a specific genetic predisposition may be necessary. The genetic mutation responsible for hereditary pancreatitis was described recently. Unlike in patients with hereditary pancreatitis, we found a lack of the R117H mutation in the cationic trypsinogen gene in all patients with tropical pancreatitis from Bangladesh.

Topics: Adolescent; Adult; Bangladesh; Chronic Disease; DNA; Female; Humans; Male; Pancreatitis; Point Mutation; Tropical Climate; Trypsinogen

The pathological activation of zymogens within the pancreatic acinar cell plays a role in acute pancreatitis. To identify the processing site where activation occurs, antibodies to the trypsinogen activation peptide (TAP) were used in immunofluorescence studies using frozen sections from rat pancreas. Saline controls or animals receiving caerulein in amounts producing physiological levels of pancreatic stimulation demonstrated little or no TAP immunoreactivity. However, after caerulein hyperstimulation (5 micrograms. kg-1. h-1) for 30 min and the induction of pancreatitis, TAP immunoreactivity appeared in a vesicular, supranuclear compartment that demonstrated no overlap with zymogen granules. The number of vesicles and their size increased with time. After 60 min of hyperstimulation with caerulein, most of the TAP reactivity was localized within vacuoles >/=1 micrometer that demonstrated immunoreactivity for the granule membrane protein GRAMP-92, a marker for lysosomes and recycling endosomes. Pretreatment with the protease inhibitor FUT-175 blocked the appearance of TAP after hyperstimulation. These studies provide evidence that caerulein hyperstimulation stimulates trypsinogen processing to trypsin in distinct acinar cell compartments in a time-dependent manner.

Topics: Acute Disease; Animals; Biomarkers; Ceruletide; Endosomes; Lysosomes; Male; Membrane Proteins; Oligopeptides; Organelles; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Time Factors; Trypsinogen; Vacuoles

Hereditary pancreatitis (HP) is the second most common cause of chronic childhood pancreatitis in the United States. Mutations in the cationic trypsinogen gene on chromosome 7 are known to cause HP. We identified four families in West Virginia with symptoms consistent with HP. To determine whether members of these families had defects in the trypsinogen gene, we tested for linkage between the HP gene and simple tandem repeat markers on chromosome 7q and screened for a specific mutation in the cationic trypsinogen gene. Two-point linkage analysis indicated that the disease gene is closely linked to three 7q markers (D7S661, D7S2511, and D7S1805). Restriction fragment length polymorphism analysis showed that all clinically affected members and nonpenetrant carriers from the four families carried a G to A mutation in the third exon of the trypsinogen gene. These findings indicate that this mutation is the cause of HP in the families in our study. The observation that most individuals who carry the mutation have symptoms of HP is consistent with the high but incomplete penetrance of the trait. The presence of a single mutation and a common linked haplotype indicates that the defective allele arose in an ancestor common to all four families.

Topics: Child; Chromosomes, Human, Pair 7; Female; Genetic Linkage; Genetic Markers; Haplotypes; Humans; Male; Mutation; Pancreatitis; Pedigree; Polymorphism, Restriction Fragment Length; Trypsinogen; West Virginia

In models of biliary acute pancreatitis, which might resemble the situation in humans, premature activation of trypsinogen inside the pancreas ("autodigestion") occurs and is correlated with the extent of ductal and parenchymal injury. It is accompanied by a critical spending of protease inhibitors and glutathione, compromising important acinar cell defense and maintenance mechanisms.. Premature activation of pancreatic digestive enzymes and profound changes of levels of certain biochemical compounds have been implicated in the pathophysiology of acute pancreatitis. Hitherto, little information on their role in biliary acute pancreatitis has been available.. Three types of injury to the pancreaticobiliary duct system of various severity were induced in rats--ligation of the common bile-pancreatic duct, retrograde infusion of electrolyte, or retrograde infusion of taurocholate solution--and were compared to sham-operated animals. Trypsin, trypsin inhibitory capacity (TIC), reduced glutathione (GSH), and other compounds were measured in pancreatic tissue. Histopathology, as well as serum amylase, lipase, and gamma-glutamyl transferase (gamma GT) were assessed.. Histopathology and elevated activity of gamma GT in the serum revealed increasing severity of pancreatic injury from sham operation through retrograde duct infusion with taurocholate. GSH was diminished even in macroscopically normal-appearing tissue, but significantly lower in altered (hemorrhagic)-looking sections. Conversely, tissue levels of trypsin were significantly increased. TIC was elevated only in the duct obstruction model, whereas it was reduced in the retrograde duct infusion models.

Topics: Acute Disease; Adenosine Triphosphate; Animals; Biliary Tract Diseases; Disease Models, Animal; Enzyme Activation; Glutathione; Humans; Male; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Serine Endopeptidases; Trypsin Inhibitors; Trypsinogen

Despite the fact that alcoholism is one of the major causes of pancreatitis, the pathogenesis of this disorder remains obscure. Factors such as the pattern of ethanol consumption, diet, and genetic predisposition may be contributing factors. The failure to produce alcoholic pancreatitis in experimental animals suggests that experimental provision of ethanol may only increase the predisposition to pancreatitis. To test this possibility, we developed an assay system using the in vitro model of cerulein-induced pancreatitis. In this system, pancreatic lobules were first exposed to a supraphysiologic concentration (10(-6) M) of the cholecystokinin analogue, cerulein, after which homogenates were incubated for up to 6 h. Activation of trypsinogen and chymotrypsinogen was observed only in cerulein-treated preparations. We then investigated the effects of the duration of ethanol feeding on cerulein-induced changes in rat pancreas. The pancreata from rats fed ethanol for 9-12 months were more susceptible to cerulein-induced activation of chymotrypsinogen compared to the pancreata from pair-fed control animals. This susceptibility also paralleled morphologic changes, such as dilatation of endoplasmic reticulum, only in the ethanol-fed group. In contrast, during the early stages (up to 3 months) of ethanol consumption, there was resistance (p < 0.01) to cerulein-induced changes. These results suggest that long-term ethanol consumption increases susceptibility to pancreatitis and raises the possibility that a similar mechanism may operate in human alcoholics.

Topics: Alcoholism; Animals; Ceruletide; Chymotrypsinogen; Endoplasmic Reticulum; Enzyme Activation; Ethanol; Male; Microscopy, Electron; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsin; Trypsinogen

The pathogenic role of acute ethanol abuse in acute pancreatitis (AP) is still obscure. The aim of the study was to evaluate the effect of antecedent intake of a high dose of 40% ethanol (5 g/kg body wt.), on trypsinogen activation, pancreatic lysosomal membrane labilization, and activities of phospholipase A2 and lipase in taurocholate AP in rats. In 80 male Wistar rats, AP or sham operation (SO) was produced 6 hr after intragastric saline (S) or ethanol (E) administration, and animals were sacrificed after 6, 12, and 18 hr. Free active trypsin (FAT) and total potential trypsin (TPT) were assayed in the pancreatic homogenate. Percentage free activity (%F/T) of cathepsin B was determined as an index of lysosomal membrane fragility. The most evident activation of trypsin occured at 6 hr AP (11.6% of TPT in S group and 16.4% in E group). Antecedent ethanol increased FAT 18 hr after SO from 0.105 +/- 0.048 microg/g protein to 0.258 +/- 0.054 and AP lasting 18 hr from 0.331 +/- 0.072 to 0.695 +/- 0.110. The %F/T of cathepsin B was highest at 18 hr of AP, suggesting maximal labilization of lysosomal membranes at this time. This labilization occurred earlier (at 12 hr of AP) in E group. The increasing effect of antecedent E on lipolytic enzymes was evident after 6 hr of AP. In conclusion, the antecedent intake of high dose of ethanol significantly promoted the conversion of trypsinogen to trypsin in taurocholate acute pancreatitis, whereas its additional effect toward labilization of pancreatic lysosomal membranes and the increase of lipolytic enzymes activities was less evident. Therefore, the promoting impact of acute ethanol intake in the development of acute pancreatitis could be mainly dependent on its increasing effect on trypsinogen activation.

Topics: Acute Disease; Animals; Cathepsin B; Ethanol; Lipase; Lysosomes; Male; Pancreatitis; Pancreatitis, Alcoholic; Phospholipases A; Phospholipases A2; Rats; Rats, Wistar; Taurocholic Acid; Trypsin; Trypsinogen

Acute pancreatitis can be difficult to diagnose. We developed a rapid dipstick screening test for pancreatitis, based on the immunochromatographic measurement of urinary trypsinogen-2.. We prospectively compared the urinary trypsinogen-2 dipstick test with a quantitative urinary trypsinogen-2 assay, a urinary dipstick test for amylase, and serum and urinary amylase assays in 500 consecutive patients with acute abdominal pain at two emergency departments. Acute pancreatitis was diagnosed according to standardized criteria.. The urinary trypsinogen-2 dipstick test was positive in 50 of the 53 patients with acute pancreatitis (sensitivity, 94 percent), including all 7 with severe pancreatitis. Two patients with urinary trypsinogen-2 concentrations below the sensitivity threshold of the test (50 ng per milliliter) and one with a very high concentration had false negative results. The test was also positive in 21 of the 447 patients without pancreatitis (specificity, 95 percent), including 7 with abdominal cancers, 3 with cholangitis, and 2 with chronic pancreatitis. The sensitivity and specificity of the dipstick test were similar to those of the quantitative urinary trypsinogen-2 assay and higher than those of the urinary amylase dipstick test. The serum amylase assay had a sensitivity of 85 percent (with a cutoff value of 300 U per liter for the upper reference limit) and a specificity of 91 percent. The sensitivity and specificity of the urinary amylase assay (cutoff value, 2000 U per liter) were 83 and 88 percent, respectively.. In patients with acute abdominal pain seen in the emergency department, a negative dipstick test for urinary trypsinogen-2 rules out acute pancreatitis with a high degree of probability. A positive test usually identifies patients in need of further evaluation.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Amylases; Female; Fluoroimmunoassay; Humans; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Prospective Studies; Reagent Strips; ROC Curve; Sensitivity and Specificity; Trypsin; Trypsinogen

Urinary TAP obtained within the first 48 h of the onset of symptoms can distinguish patients with severe acute pancreatitis.. Urinary trypsinogen activation peptide (TAP) has recently been described as an early marker of severity in acute pancreatitis.. In a multicenter study, urine samples were collected for TAP concentration at 6-12, 24, and 48 h after admission from 139 patients with acute pancreatitis (99 with mild disease, 40 with severe disease) and from 50 control patients. Severity of acute pancreatitis was defined by the presence of organ failure and/ or pancreatic necrosis on dynamic contrast-enhanced computed tomography.. Median urinary TAP in the 139 patients with acute pancreatitis compared to the 50 control patients was significantly higher at admission, 4.6 vs 0.8 ng/mL (p < 0.001), and 6-12 h, 1.9 vs 0.55 ng/mL (p = 0.04). Among patients who presented within 48 h of the onset of symptoms, the median urinary TAP for severe pancreatitis (9 patients) compared to mild pancreatitis (40 patients) was significantly higher at admission, 29.6 vs. 3.6 ng/mL (p = 0.001). Also, when obtained within 48 h of the onset of symptoms, all patients with severe pancreatitis had an admission urinary TAP level > 10 ng/mL. The sensitivity and specificity of an admission urinary TAP > or = 10 for severe pancreatitis was 100 and 85%, respectively. Given a cutoff of 10 ng/mL for an admission urinary TAP obtained within 48 h of the onset of symptoms, the negative predictive value was 100% for mild pancreatitis.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Biomarkers; Case-Control Studies; Female; Humans; Male; Middle Aged; Oligopeptides; Pancreatitis; Prognosis; Trypsinogen

The promoting effect of acute ethanol (E) abuse and protective effect of prostaglandin derivatives in acute pancreatitis (AP) remain obscure. The aim of this study was to assess the effect of previous intake of high-dose E on trypsinogen (Tn) activation and labilization of pancreatic lysosomal membranes (PLM), in taurocholate AP in rats, considering treatment with stable beta-thia-iminoprostacyclin (T). In 60 male Wistar rats taurocholate AP was induced or a sham operation was performed. Half of them received 40% E (5 g/kg body weight), 6 h earlier. T (0.3 mg/kg body weight i.g.) was applied before E or before the induction of AP. Free active (FAT) and total potential (TPT) trypsin, free (F) and total (T) cathepsin B, phospholipase A2 (PLA2), and lipase (L) activities were assayed. Percentage FAT/TPT was an index of Tn activation and fractional free (% F/T) activity of cathepsin B was an index of PLM fragility. FAT increased after 12 h of AP, and in E rats this increase was even more evident. Pretreatment and treatment with T partly prevented this increase, however, this effect was abolished or limited in rats previously given E-the changes were not effected by T. PLA2 and L activities in AP were not diminished after T. The promoting effect of acute E abuse prior to AP could be dependent on augmented activation of Tn and labilization of PLM. The protective effect of T seems to be dependent on the decrease in Tn activation in pancreatitic tissue. The potential therapeutic effect of this drug in AP could be limited by previous acute E intake, as evidenced by differences in histopathological changes.

Topics: Alcohol Drinking; Animals; Cathepsin B; Central Nervous System Depressants; Cholagogues and Choleretics; Epoprostenol; Ethanol; Lipase; Male; Pancreas; Pancreatitis; Phospholipases A; Phospholipases A2; Platelet Aggregation Inhibitors; Rats; Rats, Wistar; Taurocholic Acid; Trypsin; Trypsinogen

Topics: Chromosome Aberrations; Chromosome Disorders; Chromosome Mapping; Chromosomes, Human, Pair 7; Chronic Disease; Genes, Dominant; Genetic Markers; Humans; Pancreatitis; Trypsinogen

We recently identified a single R117H mutation in the cationic trypsinogen gene in several kindreds with an inherited form of acute and chronic pancreatitis (HP1), providing strong evidence that trypsin plays a central role in premature zymogen activation and pancreatitis. However, not all families studied have this mutation. The aim of this study was to determine the disease-causing mutation in kindreds with hereditary pancreatitis that lack the previously identified mutation.. Clinical features of the HP1 kindreds were compared with those of the new kindreds (HP2), and genetic linkage analysis, screening for mutations through DNA sequencing, and screening an unaffected population were performed.. The onset of symptoms was delayed and hospitalizations were fewer in HP2 compared with HP1 (P < 0.05). Linkage of the disease gene to chromosome 7q35 was established (logarithm of the odds, 3.73). Mutational screening identified a single A to T mutation resulting in an asparagine to isoleucine transition mutation at position 21 (N21I) in cationic trypsinogen. The mutation was absent in 94 unrelated individuals, representing 188 unique chromosomes.. The identification of a second mutation in the cationic trypsinogen gene (HP2) suggests a dominant role of trypsin in premature protease activation-mediated forms of acute pancreatitis. The pathogenesis of hereditary pancreatitis also suggests that chronic pancreatitis may result from recurrent acute pancreatitis.

Topics: Acute Disease; Adenine; Asparagine; Chromosome Mapping; Chromosomes, Human, Pair 7; Chronic Disease; Female; Genetic Linkage; Hospitalization; Humans; Isoleucine; Male; Pancreatitis; Pedigree; Point Mutation; Recurrence; Thymine; Trypsinogen

Acute pancreatitis associated with hypercalcaemia has been described in humans and experimental animals. It has been demonstrated that calcium dose dependently accelerates trypsinogen activation, and it is generally believed that ectopic activation of digestive enzymes is an early event in the pathophysiology of acute pancreatitis.. Trypsinogen activation peptide (TAP) was measured in isolated rat pancreatic acini exposed to elevated extracellular calcium in order to investigate the association between calcium and trypsinogen activation in living cells. TAP was determined in the culture medium either before (extracellular compartment) or after (intracellular compartment) cell homogenisation.. Neither secretory stimulation nor elevated calcium alone caused an increase in TAP levels. Maximal cerulein or carbachol stimulation superimposed on high medium calcium, however, significantly increased intracellular trypsinogen activation twofold. This increase was inhibited by either NG-monomethyl-L-arginine (L-NMMA) or verapamil. Acinar cell morphology and function remained intact as demonstrated by electron microscopy and secretagogue dose-response studies.. These results support the hypothesis that increased intracellular trypsinogen activation is an early step in the pathogenesis of hypercalcaemia induced pancreatitis. The model may have a bearing on other types of pancreatitis as elevated cytosolic calcium is thought to be an early event in the pathogenesis of acute pancreatitis in general.

Topics: Acute Disease; Animals; Calcium; Carbachol; Ceruletide; Culture Techniques; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Gastrointestinal Agents; Male; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

Topics: Acute Disease; Amylases; Humans; Lipase; Pancreatitis; Sensitivity and Specificity; Trypsinogen

To evaluate the clinical utility of two new tests for serum trypsinogen 2 and trypsin 2-alpha 1 antitrypsin complex (trypsin 2-AAT) in diagnosing and assessing the severity of acute pancreatitis (AP) induced by endoscopic retrograde cholangiopancreatography (ERCP).. Three hundred and eight consecutive patients undergoing ERCP at Helsinki University Central Hospital in 1994 and 1995.. Patients were followed prospectively for pancreatitis and clinical outcome. They were tested for serum trypsinogen 2, trypsin 2-AAT, and amylase in samples obtained before and one, six, and 24 hours after ERCP.. Pancreatitis developed in 31 patients (10%). Their median serum trypsinogen 2 increased 26-fold to 1401 micrograms/l at six hours after the procedure and trypsin 2-AAT showed an 11-fold increase to 88 micrograms/l at 24 hours. The increase in both markers was stronger in severe than in mild pancreatitis, and in patients without pancreatitis there was no significant increase. Baseline trypsinogen 2 and trypsin 2-AAT concentrations were elevated in 29% and 32% of patients, respectively. The diagnostic accuracy of a threefold elevation over the baseline value was therefore analysed. The sensitivity and specificity of these parameters in the diagnosis of post-ERCP pancreatitis was 93% and 91%, respectively, for serum trypsinogen 2 at six hours after the examination, and 93% and 90%, for trypsin 2-AAT at 24 hours.. Serum trypsinogen 2 and trypsin 2-AAT reflect pancreatic injury after ERCP. High concentrations are associated with severe pancreatic damage. The delayed increase in trypsin 2-AAT compared with trypsinogen 2 appears to reflect the pathophysiology of AP. A greater than threefold increase in trypsinogen 2 six hours after ERCP is an accurate indicator of pancreatitis.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; alpha 1-Antitrypsin; Amylases; Cholangiopancreatography, Endoscopic Retrograde; Female; Humans; Male; Middle Aged; Pancreatitis; Predictive Value of Tests; Prospective Studies; Statistics, Nonparametric; Trypsin; Trypsinogen

Topics: Animals; Cats; Guinea Pigs; Hypercalcemia; Pancreatitis; Rats; Trypsinogen

Ligation of the common bile-pancreatic duct induces hyperamylasemia and acute pancreatitis in rats. Pancreatic morphologic changes include edema, acinar cell damage, and mild inflammation. The pathogenesis of acute pancreatitis in this model is not understood, but may involve altered secretion and intrapancreatic activation of acinar proteases. We hypothesized that trypsinogen activation, measured by the production of plasma and pancreatic trypsinogen activation peptides (TAP), occurs early in this model. We performed the following experiments: rats were prepared with (1) bile-pancreatic ducts ligated and (2) ducts dissected but not ligated (sham). Rats were killed after 6, 24, and 48 hr. Serum amylase was measured and histologic sections were analyzed for morphologic changes. TAP was measured in both serum and pancreatic tissue homogenates using a specific polyclonal. anti-TAP antibody in an enzyme-linked immunosorbant assay. After 6, 24, and 48 hr of bile-pancreatic duct ligation, hyperamylasemia and acute morphologic changes of acute pancreatitis were observed. Evidence of acinar cell destruction was not evident until 48 hr after ligation. Levels of serum and pancreatic tissue TAP were significantly elevated at both 24 and 48 hr after ligation compared to those of sham. We conclude that increased intrapancreatic trypsinogen activation occurs early in this form of experimental acute pancreatitis and that it occurs prior to evidence of acinar cell destruction. These data and observations support the possibility that intrapancreatic protease activation contributes to the pathogenesis of ligation-induced acute pancreatitis.

Topics: Acute Disease; Animals; Ligation; Male; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsinogen

We examined the clinical utility of urine trypsinogen-2 as a marker of acute pancreatitis (AP). Fifty-nine patients with AP, 42 with acute abdominal diseases of extrapancreatic origin, and 63 without evidence of acute abdominal disease were studied. Urine trypsinogen-2 was determined by a time-resolved immunofluorometric assay. As reference methods we used serum trypsinogen-2, urine amylase, and serum amylase. The diagnostic accuracy of the markers was evaluated by receiver-operating characteristic (ROC) analysis. At admission, urine trypsinogen-2 differentiated patients with AP from controls with high accuracy. The area under the ROC curve (AUC) was 0.978, which was equal to that of serum trypsinogen-2 (0.998) and serum amylase (0.969) and significantly larger than that of urine amylase. For differentiation between severe and mild AP, urine trypsinogen-2 (0.730) was equal to serum trypsinogen-2 (0.721), and clearly better than amylase in serum and urine. These results suggest that determination of urine trypsinogen-2 is a useful test to detect AP and to evaluate disease severity.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Amylases; Female; Fluoroimmunoassay; Humans; Male; Middle Aged; Pancreatitis; Reference Values; Trypsin; Trypsinogen

The concentrations of trypsinogen-1 and -2 in serum samples from patients who have undergone pancreatectomy were measured by highly sensitive and specific time-resolved immunofluorometric assays. The isoenzyme pattern was determined by ion-exchange chromatography and determination of immunoreactivity in the fractions. All samples contained trypsinogen-2, the mean level being one fifth of that in healthy controls. Trypsinogen-1 was detected in one of nine samples. In addition to the main trypsinogen isoenzymes, we observed in normal serum two trypsinogen isoenzymes previously found in mucinous ovarian cyst fluid. Our results suggest that trypsinogen is not exclusively expressed by the pancreas and certain tumors but that it also may be produced by normal extrapancreatic tissues. This should be considered when an assay of trypsinogen in serum is used for clinical purposes.

Topics: Adenocarcinoma; Adult; Aged; Chromatography, Ion Exchange; Colonic Neoplasms; Culture Media, Conditioned; Female; Humans; Immunoradiometric Assay; Isoenzymes; Male; Middle Aged; Osmolar Concentration; Ovarian Cysts; Pancreatectomy; Pancreatitis; Postoperative Period; Reference Values; Trypsinogen; Tumor Cells, Cultured

Acute pancreatitis is characterized by hyperamylasemia, pancreatic edema, and the presence of activated digestive enzymes within the pancreas. The secretagogue-induced model of acute pancreatitis is also characterized by pancreatic acinar cell vacuolation, subcellular redistribution of lysosomal hydrolases, and a fall in pancreatic glutathione levels. We have performed time-dependence studies to determine the sequence with which these phenomena appear and to establish their cause-and-effect relationship. Evidence of lysosomal enzyme redistribution and trypsinogen activation within the pancreas could be detected within 10-15 min of the onset of supramaximal secretagogue stimulation, while hyperamylasemia (30 min), pancreatic edema (60 min), and acinar cell vacuolation (60 min) occurred at later times. Pancreatic glutathione levels were either unchanged (15 and 30 min) or elevated (60 min) during the early times of supramaximal stimulation and were only noted to be decreased at a later time. These results support the conclusion that intrapancreatic digestive enzyme activation, possibly occurring by a mechanism involving lysosomal hydrolase redistribution, is an early and likely a critical event in the evolution of secretagogue-induced pancreatitis but that glutathione depletion is neither early nor critical to the evolution of this model of pancreatitis.

Topics: Amylases; Animals; Cathepsin B; Ceruletide; Edema; Enzyme Activation; Glutathione; Male; Pancreas; Pancreatic Diseases; Pancreatitis; Rats; Rats, Wistar; Subcellular Fractions; Trypsinogen

Acute pancreatitis following endoscopic retrograde cholangiopancreatography (ERCP) occurs in 3-18% of patients undergoing either diagnostic or therapeutic ERCP. We prospectively measured urinary trypsinogen activation peptides (TAP) by an automated anti-TAP enzyme-linked immunoassay among 107 patients 4 h after ERCP to determine whether this measurement helps in the early diagnosis of ERCP-induced pancreatitis. Pancreatitis was documented in 10 of 107 patients (9.3%). All episodes were graded as mild. Urinary TAP was not significantly increased. We conclude that measurement of urinary TAP 4 h after ERCP is not helpful in documenting mild ERCP-induced acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Case-Control Studies; Female; Humans; Male; Middle Aged; Oligopeptides; Pancreatitis; Prospective Studies; Trypsinogen

Hereditary pancreatitis (HP) is a rare, early-onset genetic disorder characterized by epigastric pain and often more serious complications. We now report that an Arg-His substitution at residue 117 of the cationic trypsinogen gene is associated with the HP phenotype. This mutation was observed in all HP affected individuals and obligate carriers from five kindreds, but not in individuals who married into the families nor in 140 unrelated individuals. X-ray crystal structure analysis, molecular modelling, and protein digest data indicate that the Arg 117 residue is a trypsin-sensitive site. Cleavage at this site is probably part of a fail-safe mechanism by which trypsin, which is activated within the pancreas, may be inactivated; loss of this cleavage site would permit autodigestion resulting in pancreatitis.

Topics: Arginine; Chromosomes, Human, Pair 7; DNA Mutational Analysis; Enzyme Activation; Exons; Female; Genes; Heterozygote; Humans; Male; Models, Molecular; Pancreatitis; Pedigree; Point Mutation; Polymorphism, Restriction Fragment Length; Protein Conformation; Protein Structure, Tertiary; Trypsin; Trypsinogen

Topics: Acute Disease; Animals; Male; Pancreas; Pancreatitis; Rats; Rats, Wistar; Taurocholic Acid; Time Factors; Trypsin; Trypsinogen

The purpose of this study was to compare a time course of ultrastructural changes of secretory compartment of acinar cells in the pancreas, and a pattern of trypsinogen activation during the course of taurocholate acute pancreatitis (AP) in rats. Acute pancreatitis was induced in 21 rats by injection 0.2 ml of 5% natrium taurocholate into the biliopancreatic duct. Control rats (n = 18) were sham operated (SO). The ultrastructural and biochemical (trypsinogen activation, free active [FAT] and total potential trypsin [TPT]) examinations were performed after 6, 12 and 18 h of AP or SO. Ultrastructural lesions of acinar cells comprised of disorganization of RER, enlargement of Golgi apparatus, changes in size, shape and number of zymogen granules. These alterations were most conspicuous after 6 h of AP and they were associated with maximal activation of trypsinogen. Biochemical changes gradually normalized at 12 to 18 h of AP, however the morphological lesions persisted at these intervals of time.

Topics: Acute Disease; alpha-Amylases; Animals; Cytoplasmic Granules; Enzyme Precursors; Male; Organelles; Pancreas; Pancreatitis; Rats; Rats, Wistar; Taurocholic Acid; Trypsinogen

Dextrans improve pancreatic microcirculation in acute experimental pancreatitis. They could therefore facilitate the transport of a protease inhibitor to ischemic areas of tissue injury and be of additional benefit.. To compare the effects of dextrans with and without aprotinin, necrotizing pancreatitis was induced in 33 male dextran-resistant Wistar rats by intraductal infusion of low-dose glycodeoxycholic acid (10 mmol/l) followed by intravenous cerulein (5 micrograms/kg/h) for 6 h. Three and four hours after the start of the cerulein infusion the animals received infusions of either Ringer's lactate (RL) (12 ml/kg), 70,000 Da dextran (10%) (DEX-70) (4 ml/kg) alone, or DEX-70 (4 ml/kg) with aprotinin (5000 IU/kg) (DEX-70/A).. The death rate was 60% within 9 h in the RL group (6 of 10) but only 10% in the DEX-70 group (1 of 10) (p < 0.03; Fisher's exact test) and 23% in the DEX-70/A group (3 of 13). Histomorphometry demonstrated a significant reduction of acinar necrosis in both treatment groups compared with control animals (p < 0.014; t test). Total amounts of trypsinogen activation peptides (TAP) in ascites were also significantly lower in these groups (p < 0.05; t test).. DEX-70 given 3 h and 4 h after induction of pancreatitis significantly reduced the levels of TAP, limited acinar necrosis, and improved survival rate in acute necrotizing rodent pancreatitis. There was no additional benefit from the combination with aprotinin.

Topics: Acute Disease; Animals; Aprotinin; Ceruletide; Dextrans; Glycodeoxycholic Acid; Hemodilution; Male; Necrosis; Oligopeptides; Pancreas; Pancreatitis; Plasma Substitutes; Rats; Rats, Wistar; Serine Proteinase Inhibitors; Time Factors; Trypsinogen

The potential of pancreatic ischemia to cause acute pancreatitis as indicated by morphologic changes and ectopic trypsinogen activation was investigated.. Experimental evidence has shown that pancreatic ischemia is important in the evolution of severe pancreatitis, but whether ischemia can initiate pancreatitis has been disputed.. Pancreatic ischemia was induced in rats by hemorrhagic hypotension (30 mm Hg for 30 min; n = 64). Changes of pancreatic microcirculatory perfusion were studied using diffuse reflectance spectroscopy. Serum amylase, trypsinogen activation peptide (TAP) in serum and pancreatic tissue, wet/dry weight ratio, and histology were determined over 24 hours and compared with sham-operated control subjects (n = 35).. In control animals, serum amylase (47.9 +/- 2.1 units/L), serum (7.9 +/- 0.7 nmol/L) and tissue TAP (63.0 +/- 5.4 nmol/L x g), wet/dry weight ratio (2.8 +/- 0.1), and histology remained unchanged. Temporary hypotension markedly decreased pancreatic perfusion with incomplete recovery after reperfusion. Pancreatic isoamylase activity increased within 1 hour (110 +/- 5 units/L, p < 0.05) and further to 151 +/- 18 units/L at 24 hours. Tissue TAP was elevated at 1 hour (134 +/- 16 nmol/L x g, p < 0.05) and increased to 341 +/- 43 nmol/L x g (p < 0.001) after 24 hours, whereas serum TAP remained unchanged (8.3 +/- 0.5 nmol/L). Morphologic alterations included elevated wet/dry weight ratio (4.1 +/- 0.3, p < 0.01) and increased histologic scores for edema (p < 0.05) and acinar necrosis (p < 0.05) at 24 hours. Trypsinogen activation preceded the development of pancreatic necrosis.. In addition to its potentiating role, severe pancreatic ischemia can play a pathogenetic role in the initiation of acute pancreatitis.

Topics: Amylases; Animals; Ischemia; Isoamylase; Male; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Time Factors; Trypsinogen

It has been suggested that the severity of an attack of acute pancreatitis is related to the presence of intraglandular trypsinogen activation and that disease severity is also reflected by the degree of the acute-phase protein response. In this study we examine the relationships among amylase release, the degree of trypsinogen and prophospholipase A2 activation [as measured by urinary trypsinogen activation peptide (TAP) and prophospholipase A2 activation peptide (PLAP) concentrations], and the serum concentrations of the acute phase-protein C-reactive protein (CRP) and the principal mediator of the acute-phase protein response, interleukin-6 (IL-6). Twenty-four patients (14 mild and 10 severe attacks) were studied. Peak serum amylase concentrations were seen within 12 h and peak urinary TAP/creatinine (Cr) and PLAP/Cr ratios between 12 and 24 h after the onset of symptoms, preceding those of IL-6 and CRP. The integrated TAP/Cr and PLAP/Cr responses were significantly greater in those with severe disease [95% confidence internal (CI) = 106-259.6 pmol/mmol/h, p < 0.0008; and 95.1% CI = 462.2-3887 pmol/mmol/h, p < 0.003, respectively]. The integrated amylase response was not significantly greater in those with severe disease (95.6% CI = -415 to 832 IU/L/h, p < 0.14). There was a strong correlation among the integrated IL-6, TAP/Cr (r = 0.63, p < 0.01), and PLAP/Cr (r = 0.64, p < 0.01) responses but a poor correlation with the integrated amylase response (r = 0.19, NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acute Disease; Adult; Aged; Amylases; C-Reactive Protein; Enzyme Activation; Female; Humans; Interleukin-6; Male; Middle Aged; Oligopeptides; Pancreas; Pancreatitis; Phospholipases A; Proteins; Trypsinogen

Clinical and experimental observations have associated acute and chronic hypercalcemia with pancreatitis. The aim of this study was to determine whether acute hypercalcemia can induce acute pancreatitis and, if so, whether the pathogenesis involves premature protease activation.. Rats given bolus infusions of CaCl2 (200 mg/kg; n = 76) were compared with saline-treated controls (n = 40). Serum [Ca2+], serum amylase activity, trypsinogen activation peptide (TAP) concentration in serum and pancreatic tissue, pancreatic wet/dry weight ratio, and histology were assessed for 24 hours. For dose-response analysis, CaCl2 was injected at a dose of 50-200 mg/kg, and the aforementioned indices were assayed for 1 hour (n = 5 each).. There were no significant changes in the controls. Calcium infusion increased serum [Ca2+] 3-fold after 5 minutes (P < 0.001). Within 1 hour, serum amylase (2.5-fold) and tissue TAP (3-fold) levels increased along with macroscopic and microscopic edema formation and leukocytic infiltration. The extent of the changes at 1 hour correlated with the calcium dose. Amylase and tissue TAP concentrations remained elevated until 24 hours when serum TAP concentration had increased (P < 0.001) and focal acinar necrosis became evident.. Acute experimental hypercalcemia induces dose-dependent morphological alterations characteristic of acute pancreatitis, acute hyperamylasemia, and early ectopic trypsinogen activation. This supports the pathophysiological relevance of excess calcium and offers a possible pathogenetic mechanism for its association with clinical pancreatitis.

Topics: Acute Disease; Amylases; Animals; Calcium; Calcium Chloride; Chi-Square Distribution; Dose-Response Relationship, Drug; Enzyme Activation; Hypercalcemia; Linear Models; Male; Necrosis; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsinogen

Trypsinogen activation peptide (TAP) concentration and alpha 2-macroglobulin-trypsin complex (alpha 2M-T) activity were measured in two experimental models of acute pancreatitis in rats to evaluate the significance of activation of trypsinogen in acute pancreatitis. TAP concentration and alpha 2M-T activity in serum rose significantly in trypsin-taurocholate-induced hemorrhagic acute pancreatitis, while in cerulein-induced edematous acute pancreatitis they did not rise in spite of a similar increase in immunoreactive trypsin. When rats in trypsin-taurocholate-induced pancreatitis were treated by protease inhibitor (FUT-175; nafamostat mesilate; FUT group), alpha 2M-T activity in serum was significantly lower than that in nontreated controls (mean +/- SEM, 20.8 +/- 1.43 U/L in the FUT group vs 79.1 +/- 24.5 in controls; p < 0.01). The survival rate at 24 h was significantly improved in the FUT group compared with the controls (70 vs 43%; p < 0.05). The increase in TAP concentration in the FUT group was similar to that in controls. The TAP concentration in pancreatic tissue at 24 h was significantly (p < 0.01) lower in the survival group (7.8 +/- 0.8 ng/ml) than in the lethal group (25.9 +/- 3.7 ng/ml). Activation of trypsinogen and its subsequent enzyme activity play an important role in the evolution of severe acute pancreatitis.

Topics: Acute Disease; alpha-Macroglobulins; Animals; Benzamidines; Ceruletide; Disease Models, Animal; Guanidines; Male; Oligopeptides; Pancreatitis; Proglumide; Protease Inhibitors; Rats; Rats, Wistar; Receptors, Cholecystokinin; Taurocholic Acid; Trypsin; Trypsinogen

To study the early pathogenesis of acute edematous pancreatitis in dogs, we examined the relationship of pancreatic hyperstimulation with cholecystokinin-8 (10 micrograms/kg/hr intravenously for 6 hr) to alterations in circulating pancreatic enzymes and pancreatic morphology with special reference to trypsinogen activation. Cholecystokinin-8 infusion was associated with increases in plasma amylase, lipase, trypsin-like immunoreactivity, and plasma and urine trypsinogen activation peptide. Pancreatic parenchymal swelling and interlobular and subcapsular fluid accumulations were detected ultrasonographically within 2 hr of cholecystokinin-8. Circulating trypsin-like immunoreactivity and trypsinogen activation peptide in urine reached a peak at 2 and 4 hr, respectively, then declined despite progressive increases in circulating amylase and lipase and intrapancreatic fluid. No significant changes were observed in dogs receiving a saline infusion. This study illustrates that cholecystokinin-8 induces edematous pancreatitis in dogs that is associated with a short-lived burst of trypsinogen activation.

Topics: Acute Disease; Animals; Dogs; Edema; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Female; Oligopeptides; Pancreas; Pancreatitis; Sincalide; Time Factors; Trypsinogen; Ultrasonography

Extraintestinal trypsinogen activation peptides (TAP) have been shown to correlate with severity of acute pancreatitis in humans as well as in various animal models. Ischemia superimposed on experimental pancreatitis, however, increases acinar cell injury without increasing TAP in plasma. We speculated that TAP generated in the pancreas might not reach the circulation in necrotizing pancreatitis due to decreased pancreatic perfusion. To test the hypothesis that generation of TAP in plasma is related to pancreatic perfusion and that plasma TAP may therefore underestimate acinar cell injury in necrotizing disease, we correlated TAP in pancreatic tissue and body fluids with capillary pancreatic blood flow in necrotizing and edematous pancreatitis. The ratio between necrosis and TAP in tissue was similar in both models; the ratio between TAP in plasma and tissue, however, was significantly lower in necrotizing pancreatitis, indicating that a certain amount of TAP generated in the pancreas did not reach the circulation. Decreased pancreatic perfusion found in necrotizing pancreatitis was consistent with this finding. Our data suggest that TAP in tissue is most reliable to indicate severity of acute pancreatitis, whereas plasma TAP may underestimate pancreatic injury in necrotizing disease due to decreased pancreatic perfusion.

Topics: Acute Disease; Animals; Ceruletide; Disease Models, Animal; Edema; Glycodeoxycholic Acid; Male; Microcirculation; Necrosis; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsinogen

Previous reports demonstrated that radiographic contrast medium, as used in contrast-enhanced computed tomography, increases acinar necrosis and mortality in experimental pancreatitis. The authors studied the possibility that these changes may be related to an additional impairment of pancreatic microcirculation.. Fifty Wistar rats had acute pancreatitis induced by intraductal glycodeoxycholic acid (10 mmol/L for 10 min) and intravenous cerulein (5 micrograms/kg/hr for 6 hrs). After rehydration (16 mL/kg), pancreatic capillary perfusion was quantified by means of intravital microscopy at baseline before intravenous infusion of contrast medium (n = 25) or saline (n = 25), and 30 and 60 minutes thereafter. In addition to total capillary flow, capillaries were categorized as high- or low-flow (> or < 1.6 nL/min).. Pancreatic capillary flow did not change in either high- or low-flow capillaries after saline infusion. However, contrast medium infusion induced a significant decrease of total capillary flow (p < 0.001). Analysis according to the relative flow rate revealed that this was primarily because of a significant additional reduction of perfusion in low-flow capillaries (p < 0.0001). Furthermore, complete capillary stasis was observed in 15.9 +/- 3.4% after contrast medium as compared with 3.2 +/- 1.2% after saline infusion (p < 0.006).. Radiographic contrast medium aggravates the impairment of pancreatic microcirculation in experimental necrotizing pancreatitis.

Topics: Acute Disease; Animals; Capillaries; Contrast Media; Enzyme Activation; Iopamidol; Male; Necrosis; Pancreas; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

A porcine pancreatic transplantation model was used to investigate possible protease activation in the pancreatic graft during preservation. After perfusion with Perfadex and cold ischemia for 24 h, but prior to reperfusion, activated carboxypeptidase B was demonstrated in tissue samples from the graft parenchyma with a Western blot technique, indicating that graft pancreatitis may already be initiated during the preservation phase. A higher degree of carboxypeptidase B activation was observed in grafts perfused at a pressure of 130 cm H20 than after perfusion at 70 cm H20. During reperfusion, the fraction of activated carboxypeptidase B gradually declined but was still detectable after 2 h. One group of pigs received aprotinin intravenously during reperfusion, but the protease inhibitor did not influence the degree of carboxypeptidase B activation in the biopsy specimen. Immunoblotting against cationic trypsinogen/trypsin was also performed. When activated trypsin was detectable, it never presented more than a few percent of the total amount of uncomplexed immunoreactive trypsinogen/trypsin.

Topics: Animals; Aprotinin; Blotting, Western; Carboxypeptidase B; Carboxypeptidases; Disease Models, Animal; Enzyme Activation; Organ Preservation; Pancreas Transplantation; Pancreatitis; Perfusion; Reperfusion; Serine Proteinase Inhibitors; Swine; Trypsinogen

We developed a sensitive time-resolved immunofluorometric assay (IFMA) for trypsin-2 complexed with alpha 1-antitrypsin (AAT). We used a trypsin-2-specific monoclonal antibody on the solid phase and a europium-labeled polyclonal antibody to AAT as tracer. The detection limit is 0.05 microgram/L and the range of linearity extends to 100 micrograms/L. We compared the clinical utility of the trypsin-2-AAT assay with that of free trypsinogen-2 and amylase in serum by studying 120 healthy subjects, 29 patients with acute pancreatitis, 11 with extrahepatic biliary obstruction, and 34 with acute abdominal disorders of extrapancreatic origin. In patients with acute pancreatitis the median concentration of trypsin-2-AAT in serum was 59-fold that in healthy controls, 42-fold that in patients with biliary obstruction, and 33-fold that in patients with acute abdominal disorders of extrapancreatic origin. These differences are greater than those for trypsinogen-2 (19-, 20-, and 28-fold, respectively) and amylase (5.4-, 6.5-, and 5.4-fold, respectively). Compared with the assays of free trypsinogen-2 and amylase, our assay of trypsin-2-AAT improved the clinical specificity for acute pancreatitis by eliminating false-positive results in our control groups. Increased concentrations of trypsin-2-AAT and trypsinogen-2 were also observed in patients with chronic renal failure undergoing dialysis.

Topics: Abdomen; Acute Disease; alpha 1-Antitrypsin; Amylases; Cholestasis, Extrahepatic; Chromatography, Gel; Fluorescent Antibody Technique; Humans; Pancreatitis; Time Factors; Trypsin; Trypsinogen

Pancreatic enzyme secretion is inhibited during acute pancreatitis, resulting in an increase in acinar zymogen content. Since the premature activation of zymogens has been assigned a central role in the pathogenesis of acute pancreatitis, minimizing the amount of stored zymogens might lead to less severe acute pancreatitis. Inhibition of enzyme synthesis or stimulation of enzyme secretion would result in reduction of zymogen stores. Opiates have a varying effect on pancreatic secretion, depending on the dosage, site of administration, and presence of pancreatic stimulants. The effect of opiates and acute pancreatitis on individual pancreatic enzyme synthesis is unknown. The following study was undertaken in order to examine the effects of an opiate on pancreatic enzyme secretion and synthesis during experimental acute pancreatitis. Four groups of rats were studied. Group I received cerulein (25 micrograms/kg); group II received an opiate, buprenorphine (BPN, 0.5 mg/kg); and group III received cerulein and BPN. Drugs were dissolved in gelatin/saline and injected subcutaneously. A control group (group IV) received only gelatin/saline. Rats were sacrificed 4 hr after injection, and pancreatic mass was measured. Pancreatic acini were prepared and assayed for amylase and DNA content. Amylase, trypsinogen, chymotrypsinogen and lipase synthesis, and amylase secretion were measured for 2 hr. Results showed that, compared to controls, acini of rats with AP had increased amylase content, a finding consistent with decreased in vivo amylase secretion. Total protein and individual enzyme synthesis rates were significantly lower in the acini of the rats with AP than in those of the controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acute Disease; Amylases; Animals; Buprenorphine; Ceruletide; Chymotrypsinogen; Edema; Lipase; Male; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Trypsinogen

Although intracellular protease activation is thought to be an early event in acute pancreatitis, factors determining progression from edematous to necrotizing pancreatitis are largely unknown. With enterokinase as a probe and an immunoassay quantifying free trypsinogen activation peptides (TAP), we sought evidence for the presence of interstitial trypsinogen in edematous pancreatitis and documented the effects of its ectopic activation.. Edematous pancreatitis in the rat was induced by supramaximal stimulation with cerulein (5 micrograms/kg/hr) and coupled with enterokinase infused into the pancreatic duct at 30 mm Hg. Blue dextran infusion at this pressure corroborated interstitial delivery. Rats with no stimulation, maximal physiologic stimulation (0.25 microgram/kg/hr of cerulein), or intraductal saline infusion served as controls. TAP levels measured by enzyme-linked immunosorbent assay, 6-hour survival, and histopathology were used as end points.. Intraductal enterokinase infusion alone or in combination with maximal physiologic stimulation generated only slight increases in TAP level and no or minimal pancreatic injury. In contrast, enterokinase superimposed on edematous pancreatitis (supramaximal cerulein stimulation) produced fulminant pancreatitis and rapid death of all animals within 6 hours. Pancreatic histopathology showed severe intrapancreatic hemorrhage, acinar inflammation, and necrosis. TAP levels were significantly higher in plasma (p = 0.02), urine (p = 0.05), and ascites (p < 0.001) when compared with all other groups.. In edematous pancreatitis a large pool of trypsinogen accumulates in the interstitial space. Activation of these proenzymes leads to catastrophic consequences and may underlie progression from mild to necrotizing pancreatitis.

Topics: Animals; Ceruletide; Edema; Enteropeptidase; Enzyme Activation; Male; Necrosis; Oligopeptides; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsinogen

Trypsinogen activation peptides (TAP) can be found in patients with acute pancreatitis (AP) as well as in different models of experimental AP. First experience has suggested that early elevation of TAP indicates development of necrotizing AP and that the amount of TAP correlates with the extent of acinar cell necrosis and mortality. It is however unknown whether TAP similarly assess severity of AP in the later course of the disease. The present study monitores TAP in plasma, urine and ascites during the initial development of pancreatic injury and correlates the amount of TAP and the extent of pancreatic necrosis over 48 h in a rodent model of AP. While there was no elevation of TAP in control animals or animals with edemantous AP, significant amounts of TAP im plasma were found as early as 30 min following induction of severe necrotizing AP. Serum amylase returned to normal values within 24 h, TAP maintained at high levels until the end of the 48 h observation period. During the first 24 h TAP paralleled the development of acinar cell necrosis, but decreased thereafter despite further progression of pancreatic injury. Our results provide further evidence suggesting that TAP initially precede morphological changes in AP. Early in the course of the disease TAP parallel development of pancreatic injury.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acute Disease; Animals; Male; Necrosis; Oligopeptides; Pancreas; Pancreatitis; Rats; Rats, Sprague-Dawley; Trypsinogen

Inappropriate extraluminal activation of trypsin is assumed to play a part in the pathogenesis of acute pancreatitis (AP), but proof has been elusive because active trypsin is transient and difficult to measure. We have previously shown increased levels of trypsinogen activation peptides (TAP), a direct measure of trypsin activation, to correlate with severity of AP, tissue necrosis, and survival in a rodent model induced by cerulein hyperstimulation and bile salt infusion. The present study seeks to show that increased trypsinogen activation also characterizes three other models of experimental AP in rodents to give credence to the generality of the phenomenon and to its potential relevance to human AP.. Experimental AP was induced in mice by a choline-deficient diet supplemented with ethionine and in rats by creation of a closed duodenal loop or by ligation of the biliopancreatic duct plus physiologic stimulation. TAP were quantified by an immunoassay in tissue and plasma at various time points after onset of AP.. In the group with choline-deficient diet supplemented with ethionine a significant increase in tissue and plasma TAP was found at 48 and 72 hours, respectively. In the group with closed duodenal loop significant TAP elevations were found in plasma as early as 6 hours and in the group with ligation of the biliopancreatic duct plus physiologic stimulation at 24 hours.. These experiments provide further evidence that extraluminal protease activation is a pathophysiologic event common to the evolution of various models of experimental acute pancreatitis and therefore increase the likelihood that this phenomenon is important in the human disease as well.

Topics: Acute Disease; Animals; Choline Deficiency; Diet; Disease Models, Animal; Duodenum; Enzyme Activation; Ethionine; Female; Ligation; Male; Mice; Necrosis; Oligopeptides; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Trypsinogen

Protein analysis of intraductal precipitates and calculi is important to elucidate the mechanism of stone formation in chronic pancreatitis. We revealed human cationic trypsin immunoreactivity in protein extracts of pancreatic stones from 11 of 13 patients with chronic calcified pancreatitis, ranging from 0 to 42.3 ng/micrograms protein. On gel filtration the immunoreactivity eluted as one peak, which is identical to that of human cationic trypsinogen. On immunostaining of pancreatic stone, using an immunogold technic and scanning electron microscopy, the immunoreactivity was observed more densely in the amorphous portion of the center of the stones than in the concentric laminar layer of the periphery. Only negligible activity was detected for elastase 1 or amylase in the stone extracts. These results suggest that the presence of trypsinogen in pancreatic stone is not due to coprecipitation or adsorption of pancreatic enzymes but that trypsinogen is more likely involved in an initial step of intraductal precipitate formation than in a subsequent step of stone formation. However, the absence of trypsinogen in the stones from two of the 13 patients also suggests that trypsinogen is not the sole protein initiating precipitate formation.

Topics: Adult; Aged; Calculi; Chronic Disease; Female; Humans; Immunohistochemistry; Male; Microscopy, Electron, Scanning; Middle Aged; Pancreatic Diseases; Pancreatitis; Proteins; Trypsin; Trypsinogen

Contrast-enhanced computed tomography (CECT) is used to show areas of decreased pancreatic perfusion in severe acute pancreatitis (AP). To evaluate possible adverse effects of the contrast medium (CM) on the course of AP, the impact of intravenous CM in AP of graded severity in the rat was studied.. Pancreatitis of three levels of severity was induced in Sprague-Dawley rats with intravenous cerulein hyperstimulation plus time- and pressure-controlled intraductal infusion of saline or glycodeoxycholic acid. At 7 hours, control and pancreatitis animals received intravenous ionic CM, nonionic CM, or saline. The principal outcome measures were 24-hour survival, trypsinogen activation peptides (TAP) in ascites, and histological acinar necrosis score.. There was no measurable effect of CM on the index features in control animals or animals with mild or moderate AP. In severe AP, CM caused a significant increase in mortality, ascites TAP, and necrosis score.. Intravenous CM increases pancreatic injury when administered early in the course of severe experimental AP. Because CM may convert borderline ischemia to irreversible necrosis, CECT performed early in pancreatitis to show poor perfusion and predict areas of necrosis may depict a self-fulfilling prophecy. Early CECT should be reconsidered and perhaps avoided.

Topics: Acute Disease; Animals; Ascites; Ceruletide; Contrast Media; Glycodeoxycholic Acid; Injections; Injections, Intravenous; Male; Necrosis; Pancreatic Ducts; Pancreatitis; Peptides; Rats; Rats, Sprague-Dawley; Sodium Chloride; Survival Analysis; Time Factors; Trypsinogen

Acinar necrosis in patients with acute pancreatitis can be due to enzymatic injury, ischemia, or both. We hypothesized that novel therapy aimed at an improvement of pancreatic microcirculation early in the course of pancreatitis may reduce the lethality and acinar damage. Forty-six dextran-resistant rats received controlled intraductal infusion of glycodeoxycholic acid (10 mmol/L), followed by intravenous cerulein (5 micrograms/kg/h) for 6 hours. Beginning 30 minutes after the induction of pancreatitis, all animals were resuscitated with Ringer's lactate (RL) (8 mL/kg/h intravenously for 9 hours). In addition, they were given intra-aortic bolus infusions (2 mL/kg at 30, 60, 90, and 150 minutes) of either RL, sodium chloride (NaCl) (7.5%) and dextran 60,000 (10%) (HHS-60), NaCl (7.5%) and dextran 500,000 (10%) (HHS-500), or NaCl (0.9%) and dextran 500,000 (10%) (DEX-500). Despite high-volume fluid resuscitation in the groups that received RL and HHS-60, 70% of the animals in each of these groups died within 24 hours. In contrast, the mortality rates in the groups of animals that received HHS-500 and DEX-500 were dramatically reduced to 0% and 10%, respectively (p = 0.005, p = 0.02). Histopathologic scores for acinar necrosis were significantly lower in the group of animals that received DEX-500 (p < 0.009) compared with those that received RL and HHS-60. Finally, total amounts of trypsinogen activation peptides in ascites were significantly lower in the animals that received HHS-500 (p < 0.004) and DEX-500 (p < 0.02) compared with those that received RL and HHS-60. Rapid bolus infusion of hyperoncotic ultrahigh molecular weight dextran solution with or without hypertonic saline but not RL or hypertonic-hyperoncotic saline-dextran significantly reduced pathologic trypsinogen activation, prevented acinar necrosis, and improved survival in acute experimental pancreatitis. We speculate that a sustained improvement of pancreatic microcirculation by ultrahigh molecular weight dextran is the mechanism of action.

Topics: Animals; Dextrans; Fluid Therapy; Isotonic Solutions; Male; Microcirculation; Necrosis; Pancreas; Pancreatitis; Rats; Rats, Wistar; Ringer's Lactate; Saline Solution, Hypertonic; Trypsinogen

The effects of single and repeated short-term (4 hr) obstruction of pancreaticobiliary duct (PBDO), with or without exocrine stimulation (intraductal hypertension) by cerulein infusion (0.2 micrograms/kg.hr), on the exocrine pancreas were evaluated in the rat. Single blockage of pancreaticobiliary duct for 4 hr caused a significant rise in serum amylase levels, pancreatic water content, and redistribution of lysosomal enzyme, cathepsin B from the lysosomal fraction to the zymogen fraction, which was considered to mean the colocalization of lysosomal enzymes with pancreatic digestive enzymes in the same subcellular compartment in acinar cells. In addition, the accelerated lysosomal and mitochondrial fragility was observed in the single pancreaticobiliary-duct-obstructed animals. Moreover, the repeated PBDO for 4 hr (2 hr in each obstruction and 1 hr of free flowing of pancreaticobiliary juice between two obstructions) caused more marked changes in almost the all parameters, and the repeated PBDO with intraductal hypertension caused an activation of trypsinogen in the pancreas, making more marked changes in almost the all parameters than the repeated PBDO only group. These results indicate that the present model of repeated PBDO with exocrine stimulation seems to be a pertinent model for gallstone pancreatitis in humans, and that redistribution of lysosomal enzymes and subcellular organellar fragility seem to play an important role in the pathogenesis of pancreatic injuries induced by PBDO, particularly by repeated PBDO with exocrine stimulation, probably via activation of trypsinogen to trypsin by lysosomal enzyme, cathepsin B.

Topics: Acute Disease; Amylases; Animals; Body Water; Cathepsin B; Ceruletide; Cholecystitis; Cholelithiasis; Male; Pancreas; Pancreatic Ducts; Pancreatitis; Rats; Rats, Wistar; Trypsin; Trypsinogen

The redistribution of cathepsin B, a representative lysosomal enzyme, from the lysosomal pellet to the zymogen pellet in subcellular fractions and the colocalization of cathepsin B with digestive enzymes within acinar cells have been found during the early stage of caerulein-induced acute pancreatitis in the rat. This study investigated the protective effects of prostaglandins E1 and E2 on the exocrine pancreas in this experimental pancreatitis. Prostaglandin E2, but not E1, prevented the redistribution of cathepsin B along with the hyperamylasemia, and the increase in amylase and trypsinogen in the acinar cells in almost a dose-dependent manner, particularly at a dose of 100 micrograms/kg.hr of continuous infusion. These results suggest that subcellular organelle fragility is closely related to the pathogenesis of acute pancreatitis, and that prostaglandin E2 has an important cytoprotective effect on biological membranes as a stabilizer of lysosomal membrane.

Topics: Alprostadil; Amylases; Animals; Cathepsin B; Ceruletide; Dinoprostone; Dose-Response Relationship, Drug; Lysosomes; Male; Pancreas; Pancreatitis; Rats; Rats, Wistar; Trypsinogen

Topics: Humans; Lupus Erythematosus, Systemic; Pancreas; Pancreatitis; Trypsinogen

Recent experimental findings have suggested that activation of trypsinogen by cathepsin B within acidic pancreatic acinar cell cytoplasmic vacuoles may be a critical early event in both secretagogue and diet-induced pancreatitis. The weak base chloroquine accumulates within acidic intracellular compartments, raises their pH, and can inhibit proteolysis as well as cathepsin B.. We have investigated the effect of in vivo chloroquine administration on both secretagogue and diet-induced experimental pancreatitis to determine if raising the pH of cytoplasmic vacuoles in these models of pancreatitis would have a protective effect.. Infusion of chloroquine (5 mg.kg-1.h-1) resulted in the uptake and concentration of chloroquine in the pancreas, an increase in the pH of acinar cell acidic compartments, and interference with the pH-dependent sorting of lysosomal hydrolases from digestive enzyme zymogens. However, chloroquine administration did not have a protective effect against the hyperamylasemia, the pancreatic edema, the morphological changes or the mortality that is associated with these models of pancreatitis.. These observations lead us to conclude that raising the pH of acinar cell acidic compartments by in vivo administration of chloroquine does not prevent either secretagogue or diet-induced pancreatitis.

Topics: Acute Disease; Animals; Cathepsin B; Chloroquine; Enzyme Activation; Hydrogen-Ion Concentration; Male; Pancreatitis; Rats; Rats, Wistar; Trypsinogen; Vacuoles

The redistribution of cathepsin B, a lysosomal enzyme, from the lysosomal pellet to the zymogen pellet in the subcellular fractionation, the colocalization of cathepsin B with digestive enzyme, and increased cellular, lysosomal, and mitochondrial fragility within acinar cells have been found during the early stages of caerulein-induced acute pancreatitis in rats. In the present study, the authors investigated the protective effects of prostaglandin E1 and E2, a combined therapy of these prostaglandins, and a new, synthetic, low molecular weight protease inhibitor, ONO3307, on the exocrine pancreas in this noninvasive model of experimental pancreatitis in vivo and in vitro. Prostaglandin E2, but not E1, prevented hyperamylasemia, congestion of amylase and trypsinogen in the acinar cells, redistribution of cathepsin B, and amylase and lactate dehydrogenase discharge from the dispersed acini. It also prevented cathepsin B leakage from the lysosomes and malate dehydrogenase leakage from the mitochondria in an almost dose-dependent manner, particularly at the dose of 100 micrograms/kg/hr continuous infusion. Furthermore, the combined therapy of prostaglandin E2 with ONO3307 strongly inhibited all the parameters tested in this study. This combination therapy seems to be the most effective against secretagogue-induced pancreatic injuries. These results indicate that cellular and subcellular organellar fragility seem to be closely involved in the pathogenesis of acute pancreatitis. Prostaglandin E2 seems to have important cytoprotective effects on the biologic membranes, such as a stabilizer of lysosomal or mitochondrial membranes. In addition, these findings also suggest the crucial roles of some unknown proteases in the etiology of acute pancreatitis, and indicate the clinical effectiveness of prostaglandins and this type of low molecular weight protease inhibitor for acute pancreatitis.

Topics: Acute Disease; Alprostadil; Amylases; Animals; Body Water; Cathepsin B; Ceruletide; Dinoprostone; Guanidines; L-Lactate Dehydrogenase; Lysosomes; Male; Microsomes; Pancreas; Pancreatitis; Rats; Rats, Wistar; Serine Proteinase Inhibitors; Subcellular Fractions; Trypsinogen

In rats fed control and ethanol-containing Lieber-DeCarli diets for a period of 12 months, the bile did not contain any enterokinase, the pancreatic juice did not contain any plasmin or thrombin, but in animals fed high fat diet with ethanol, trypsinogen and chymotrypsinogen were significantly increased and trypsin inhibitor decreased. In the tissue, free trypsin and cathepsin B were increased. Composite profile of trypsinogen in gel segments obtained from the pancreatic juice and the tissue showed higher peaks of cationic and anionic variants of trypsinogen in animals fed ethanol. There was no evidence of mesotrypsinogen or of enzyme Y in the juice or the tissue. These studies show that serine proteases and cathepsin B may play a major role in the pathobiology of alcoholic pancreatitis.

Topics: Alcoholism; Animals; Bile; Chymotrypsinogen; Dietary Fats; Enzyme Activation; Ethanol; Pancreas; Pancreatic Juice; Pancreatitis; Rats; Trypsinogen

Trypsinogen activation peptides (TAP) were quantified by radioimmunoassay in blood, urine, and peritoneal exudate of rats with experimental pancreatitis. Forty-four animals were studied, comprising a control group and four different induction techniques (cerulein, cerulein plus either 2- or 10-min intraductal glycodeoxycholic acid [GDOC] infusion, and cerulein plus intraductal GDOC with enterokinase [EK]). Significantly higher TAP concentrations were found at 6 h (or at death) in plasma and ascites of all pancreatitis groups compared with controls. TAP quantitation in hourly urine samples demonstrated significantly higher concentrations from the third hour onward in the most severe groups and from the fourth hour onward in the cerulein-treated rats. All nonsurviving rats had a plasma TAP of greater than 2.5 nM/L, whereas only 1 of 34 surviving animals had such a concentration (p less than 0.001). A significant stepwise increase in total TAP in ascites was found when comparing the cerulein group, the two GDOC groups, and the EK group (p less than 0.001). Chromatography of samples with a high TAP content demonstrated comigration with synthetic TAP. We conclude that free TAP are present in blood, urine, and peritoneal exudate of rats with experimental pancreatitis of different pathogenesis and that the amount of TAP may be indicative of the severity of the disease process.

Topics: Acute Disease; Amino Acid Sequence; Amylases; Animals; Ascites; Male; Molecular Sequence Data; Pancreatitis; Peptide Biosynthesis; Peptides; Rats; Rats, Inbred Strains; Trypsinogen

We studied the effect of short-term (3 h) pancreatic duct obstruction (PDO) on the exocrine pancreas and on the secretion of lysosomal enzymes into the pancreatic juice of rabbits during stimulation by pancreatic secretagogues. The following evaluations were made: serum amylase levels, pancreatic water content, pancreatic amylase, trypsinogen and cathepsin B content, and output of pancreatic enzymes and lysosomal hydrolases when stimulated by secretin and caerulein as well as the distribution of cathepsin B in subcellular fraction. Cellular fragility (LDH leakage from dispersed acini) and subcellular organellar fragility (cathepsin B leakage from lysosomes and malate dehydrogenase leakage from mitochondria) were also evaluated. PDO for 3 h plus secretin infusion caused a significant rise in serum amylase levels, pancreatic water content, and pancreatic amylase and trypsinogen content due to congestion of digestive enzymes during PDO. There was also a redistribution of cathepsin B from the lysosomal fraction to the zymogen fraction and increased cellular and subcellular organellar fragility. In normal rabbits and in those with only secretin infusion, caerulein stimulated the secretion of cathepsin B into pancreatic juice. Just after PDO, the secretion of cathepsin B, amylase and trypsinogen significantly decreased. By 24 h after PDO, the output of cathepsin B stimulated by caerulein and secretin had increased significantly. Amylase and trypsinogen output were also significantly increased at this stage, in both the secretin and caerulein fractions. These results indicate that the secretion of lysosomal enzymes into pancreatic juice is stimulated by gut hormones, such as caerulein, in the normal physiological state and in pathological states, such as PDO. These results also show an important role of increased cellular and subcellular organellar fragility in the pathogenesis of pancreatic injuries induced by PDO and augmented secretion of both lysosomal enzymes and pancreatic digestive enzymes in the recovery stage after PDO and their important roles at this stage. Lysosome enzymes also seem to play some physiological roles in the pancreatic ductal system in normal physiological states as well as their roles in pathological states, because cathepsin B can activate trypsinogen, and trypsin can activate many other enzymes.

Topics: Amylases; Animals; Cathepsin B; Ceruletide; L-Lactate Dehydrogenase; Lysosomes; Male; Pancreas; Pancreatic Ducts; Pancreatitis; Rabbits; Secretin; Trypsinogen

Intrapancreatic activation of trypsinogens is believed to occur either as a cause or a consequence of acute pancreatitis and to be associated with the more severe forms of the disease. Trypsinogen-activation peptides (TAPs) were measured in plasma, urine, and ascites of rats (n = 54) assigned to different pancreatitis-inducing regimens reproducing the entire spectrum of severity. Compared with survivors, nonsurvivors at 9 hours had significantly higher TAP levels in plasma at 3 hours (P = 0.0001), urine (peak, 1-4 hours) (P = 0.004), and ascites (P = 0.0001) after death. Stepwise discriminant analysis showed that TAP in urine and plasma were the most accurate predictors of outcome (88.2% of animals) compared with other routine laboratory parameters. Morphometric analysis showed that the best histopathologic correlates of TAP elevation were acinar necrosis and intrapancreatic hemorrhage. In a second series of experiments using a homogeneous technique of induction producing pancreatitis with a mortality of 55% at 48 hours, plasma TAP level at 3 hours (cutoff, 0.5 nmol/L) and/or urinary TAP level (peak, 1-6 hours; cutoff, 25 nmol/L) accurately predicted outcome in 85% of animals. It is concluded that the TAP assay gives an accurate early prediction of outcome in different pancreatitis models and correlates best with acinar necrosis and hemorrhage.

Topics: Acute Disease; Animals; Ascites; Glycodeoxycholic Acid; Male; Pancreatitis; Peptides; Prognosis; Rats; Rats, Inbred Strains; Severity of Illness Index; Trypsinogen

Topics: Cardiopulmonary Bypass; Enzyme Activation; Humans; Hypercalcemia; Pancreatitis; Trypsinogen

We investigated pancreatic gene expression in the rat in response to taurocholate-induced acute pancreatitis. Concentrations of transcripts encoding pancreatic protein showed noncoordinated alterations. Contents in amylase, trypsinogen I, chymotrypsinogen B, elastase 1, and procarboxypeptidase A mRNAs decreased by greater than 50% during the acute phase (days 0-2), whereas actin and lithostathine mRNAs increased 5 and 0.6 times, respectively, and pancreatitis-associated protein (PAP) mRNA increased greater than 200 times, indicating redirection of the pattern of gene expression. Synthesis of pancreatic proteins was also altered in a noncoordinated manner. During the acute phase, it decreased more for trypsinogen I and chymotrypsinogen B than for amylase and lipase, whereas synthesis of the PAP increased dramatically. For amylase and chymotrypsinogen B, we compared the patterns of changes in mRNA concentrations, rates of synthesis, and pancreatic contents. Changes in enzyme contents and synthetic rates were temporally correlated during the acute phase. On the contrary, changes in mRNA concentrations and enzyme synthesis were not coordinated, suggesting that control of synthesis partly occurred at the posttranscriptional level. It was concluded that induction of pancreatitis is accompanied by transcriptional and posttranscriptional modifications resulting in rapid and massive rearrangement of the pattern of pancreatic protein gene expression.

Topics: Acute Disease; Amylases; Animals; Carboxypeptidases; Carboxypeptidases A; Chymotrypsinogen; DNA Probes; Enzyme Precursors; Gene Expression; Lipase; Male; Methionine; Pancreas; Pancreatitis; Pancreatitis-Associated Proteins; Rats; Rats, Inbred Strains; RNA, Messenger; Taurocholic Acid; Trypsinogen

A preliminary investigation of the role of ultrasound, including color and duplex Doppler, was performed in recipients of cadaveric pancreatico-duodenal transplants. Twenty such examinations were done on three patients. Three different complications were noted: rejection, pancreatitis, and peripancreatic abscess. The mean normal resistive index (RI) was 0.71 +/- 0.12. The normal allograft anteroposterior (AP) dimension ranged from 1.5 to 2.0 cm. Intraparenchymal and main feeding vessels were demonstrated easily. RI calculations alone were not helpful in diagnosing graft rejection. However, this diagnosis can be made using a new biochemical marker, serum anodal trypsinogen. We conclude that when used in conjunction with a reliable biochemical marker for rejection (serum anodal trypsinogen), ultrasound, including color and duplex Doppler, provides an important adjunct for the rapid, inexpensive, and complete evaluation of patients with pancreatico-duodenal transplants.

Topics: Cadaver; Diabetes Mellitus, Type 1; Duodenum; Graft Rejection; Humans; Pancreas Transplantation; Pancreatitis; Postoperative Complications; Trypsinogen; Ultrasonics; Ultrasonography

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Female; Humans; Male; Middle Aged; Pancreatitis; Peptides; Prognosis; Prospective Studies; Radioimmunoassay; Trypsinogen

Trypsinogen activation can be quantified by measurement of released activation peptides (TAP assay). TAP assay in urine was performed on admission for 55 patients with acute pancreatitis. TAP concentration correlated with subsequent disease severity in 87%, whereas C-reactive protein concentration, and multifactorial scoring at 48 h, were correct in 55% and 84%. Sensitivity and specificity for TAP assay were 80% and 90%, for C-reactive protein 53% and 55%, and for multifactorial scoring at 48 h, 60% and 93%. Urine TAP assay distinguishes acute pancreatitis without trypsinogen activation from acute pancreatitis with trypsinogen activation, and helps to identify patients who will progress to the severe acute disease. Use of the assay should allow early intensive treatment of those who need it.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; C-Reactive Protein; Evaluation Studies as Topic; Female; Humans; Length of Stay; Male; Middle Aged; Pancreatic Polypeptide; Pancreatitis; Peptides; Predictive Value of Tests; Prognosis; Radioimmunoassay; Time Factors; Trypsinogen

Investigations of pancreatic juice revealed new insights into the pathogenesis of chronic pancreatitis (cP). But many results are contradictory. In this paper pure human pancreatic juice from patients with cP (n = 14) was compared with results obtained from normal subjects (n = 22). The pancreatic juice was obtained endoscopically recording the absorption (280 nm) simultaneously. By means of this special technique 4 fractions could be exactly distinguished: 1. wash-out-period, 2. phase of secretin action, 3. phase of pancreozymin (CCK) action, and 4. post-CCK-phase. Total protein, trypsinogen, zinc sodium, and potassium were determined. In fraction 1 (wash-out-period) mean values of protein, trypsinogen and zinc are lower in patients with cP compared with control subjects. In case of zinc the difference is statistically significant. In fraction 2 (secretin-phase) no differences could be detected between cP and control subjects. In contrast in fraction 3 (CCK-phase) mean values of protein and trypsinogen are lower in control subjects than in patients with cP. But the standard deviations are so high that all differences are not statistically significant. The results indicate that under fasting conditions the pancreatic juice content of protein, trypsinogen and zinc is lower in patients with cP. But patients with cP can be stimulated much better with CCK than control subjects. Till now such a different behaviour during wash-out-period and CCK-stimulation is not reported in the literature.

Topics: Cholangiopancreatography, Endoscopic Retrograde; Cholecystokinin; Chronic Disease; Humans; Pancreatic Function Tests; Pancreatic Juice; Pancreatitis; Potassium; Proteins; Secretin; Sodium; Trypsinogen; Zinc

Intracellular localization and enzymatic activities of lysosomal enzymes (cathepsin B, N-acetyl-beta-glucosaminidase, and beta-glucuronidase) were studied in control rats and after induction of caerulein pancreatitis. In control rats high enzymatic activities were found in the postnuclear 1000 g fraction (purified zymogen granules). The corresponding subcellular fraction in pancreatitis animals additionally contained larger secretory vacuoles and autophagosomes and revealed a marked increase in lysosomal enzyme activities. Immunolabelling studies at the ultrastructural level for trypsinogen and cathepsin B demonstrated a colocalization of lysosomal and digestive enzymes in zymogen granules in healthy controls. After induction of pancreatitis immunolabelling still demonstrated a colocalisation of cathepsin B and trypsinogen in secretory granules and newly formed Golgi-derived secretory vacuoles. Concomitantly appearing autophagosomes were, however, only labelled for cathepsin B. It is concluded that segregation of lysosomal and digestive enzymes is incomplete in normal acinar cells resulting in a colocalization in zymogen granules. In pancreatitis colocalization in secretory granules is maintained, whereas only lysosomal enzymes were sufficiently transferred into autophagic vacuoles. No indication for impaired mechanisms of molecular sorting of lysosomal and digestive enzymes in caerulein-induced pancreatitis was found.

Topics: Animals; Ceruletide; Cytoplasmic Granules; Enzyme Precursors; Hydrolases; Immunohistochemistry; Lysosomes; Male; Pancreatitis; Rats; Rats, Inbred Strains; Trypsinogen

We have developed sensitive time-resolved immunofluorometric assays for the two trypsinogen isoenzymes, trypsinogen-1 and trypsinogen-2, which also are called cationic and anionic trypsinogen, respectively. The assays use monoclonal antibodies produced by immunization with tumor-associated trypsinogen that is isolated from mucinous ovarian cyst fluid. In each assay, one antibody is immobilized onto the walls of polystyrene microtiter strip wells and the other is labeled with an europium(III) chelate. The cross-reaction of each trypsinogen isoenzyme in the assay for the other isoenzyme is less than 1%. The detection limits are 0.1 micrograms/L for trypsinogen-1 and 0.3 micrograms/L for trypsinogen-2. In sera of healthy subjects and patients with extrapancreatic disease the concentration of trypsinogen-1 is higher (median, 21 micrograms/L) than that of trypsinogen-2 (median, 17 micrograms/L), but in acute pancreatitis the ratio is reversed. In acute pancreatitis the concentration of trypsinogen-2 is 50-fold higher than in controls, whereas the difference in trypsinogen-1 concentrations is only 15-fold. The corresponding difference in immunoreactive trypsin measured by a commercially available radioimmunoassay was also only 10-fold.

Topics: Acute Disease; Adult; Aged; Antibodies, Monoclonal; Biomarkers; Cross Reactions; Female; Fluoroimmunoassay; Humans; Isoenzymes; Male; Middle Aged; Ovarian Neoplasms; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

The optimal strategy for the diagnosis of acute pancreatitis with enzyme assay results as indicators was evaluated in 67 emergency cases in whom this condition was suspected. We measured urine amylase expressed as activity concentration (U/L), timed excretion (U/h), and amylase/creatinine clearance ratio, and also serum amylase, elastase, lipase, and trypsinogen, at admission and repeatedly during hospitalization. The receiver-operator characteristic function was used to evaluate the diagnostic discrimination of each variable among initial findings and among the highest individual findings established retrospectively. We applied the same treatment to multiple univariate discrimination, using the six possible pairs and the four possible triplets of serum indicators. The results suggest that such urine assays should be abandoned, that all individual serum assays combine about 0.9 sensitivity with 0.9 specificity, that pairing of two assays does not clearly enhance discrimination, and that triplets of tests may degrade discrimination. The trade-off between sensitivity and specificity is a function not only of the chosen decision threshold but also of the sampling strategy (initial vs highest values) and of the interpretation rule (Boolean "and" vs "or" strategy).

Topics: Acute Disease; Amylases; Clinical Enzyme Tests; Creatinine; False Negative Reactions; False Positive Reactions; Female; Humans; Lipase; Male; Pancreatic Elastase; Pancreatitis; Trypsinogen

The activation of zymogen proteases and lysosomal enzyme cathepsin B in the pancreas was investigated in cerulein-induced pancreatitis in rats. Acute pancreatitis was induced by two intraperitoneal injections of 40 micrograms/kg of body weight of cerulein at intervals of 1 h. After the first cerulein injection, the active trypsin and elastase contents in the pancreas tissues significantly increased, and reached the highest level at 3 h after the first injection, followed by peaks at 5 h in the serum amylase and lipase levels and the pancreas wet weight. Cathepsin B contents in pancreas tissues showed a parallel increase with active zymogen enzymes during the first 3 h of pancreatitis. These findings may suggest that the intracellular activation of trypsinogen is an important step in the development of cerulein-induced acute pancreatitis and that cathepsin B plays a role in the activation of trypsinogen in pancreatic acinar cells.

Topics: Amylases; Animals; Body Weight; Cathepsin B; Ceruletide; Enzyme Precursors; Lipase; Lysosomes; Male; Pancreas; Pancreatic Elastase; Pancreatitis; Peptide Hydrolases; Rats; Rats, Inbred Strains; Trypsin; Trypsinogen

A study on the effect of zinc feeding on the survival rate as well as the levels of trypsinogen, alpha 2-macroglobulin, zinc, calcium, and magnesium in the plasma, pancreata, and livers of BALB/c mice fed a choline-deficient diet supplemented with 0.5% DL-ethionine (CDE diet) was undertaken. Feeding them a zinc-excess diet significantly increased the survival rate of mice with pancreatitis induced by CDE diet feeding. Trypsinogen concentrations in plasma and pancreas increased in mice fed a CDE diet and further increased in mice fed a zinc-deficient diet. The plasma alpha 2-macroglobulin levels in mice fed a zinc-deficient diet decreased compared to those fed a zinc-adequate or a zinc-excess diet. In mice with pancreatitis, zinc and calcium concentrations of pancreata increased and magnesium concentrations decreased compared to those of normal controls. The calcium concentrations in both livers and pancreata increased, but magnesium concentrations in these tissues decreased. These results suggest that altered mineral metabolism in the pancreas may have contributed to the pathophysiology of the mice with acute pancreatitis and that zinc supplementation in the diet may be therapeutic for pancreatitis.

Topics: Acute Disease; alpha-Macroglobulins; Animals; Calcium; Choline Deficiency; Diet; Female; Magnesium; Male; Mice; Mice, Inbred BALB C; Pancreatitis; Survival Rate; Trypsinogen; Zinc

In the present study fine structural changes of acinar zymogen granules were investigated in human acute pancreatitis. Pancreatic tissue was obtained at surgery from 6 patients, prepared for ultrastructural analysis, and stained immunocytochemically for trypsinogen. Stereological parameters of zymogen granules were evaluated. The density of the immunocytochemical labelling for trypsinogen was estimated over zymogen granules, the rough endoplasmic reticulum, Golgi apparatus and the acinar lumina. In acute pancreatitis the number of zymogen granules was diminished and their size reduced. The density of the labelling for trypsinogen was unchanged over zymogen granules but showed a significant reduction over the rough endoplasmic reticulum, Golgi apparatus, and the acinar lumina. In general the integrity of zymogen granules was well preserved. Focally degenerative changes of zymogen granules and large autophagosomes were observed. From the immunogold labelling a disturbance of enzyme synthesis and secretion was suggested. Evidence is given that a disruption of the zymogen granule membranes and a fusion with lysosomal bodies might contribute to the pathogenesis of human acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Cathepsin B; Cytoplasmic Granules; Enzyme Precursors; Humans; Immunohistochemistry; Microscopy, Electron; Middle Aged; Pancreas; Pancreatitis; Trypsinogen

The pancreatic stone protein and its secretory form (PSP-S) are inhibitors of CaCO3 crystal growth, possibly involved in the stabilization of pancreatic juice. We have established the structure of PSP-S mRNA and monitored its expression in chronic calcifying pancreatitis (CCP). A cDNA encoding pre-PSP-S has been cloned from a human pancreatic cDNA library. Its nucleotide sequence revealed that it comprised all but the 5' end of PSP-S mRNA, which was obtained by sequencing the first exon of the PSP-S gene. The complete mRNA sequence is 775 nucleotides long, including 5'- and 3'- noncoding regions of 80 and 197 nucleotides, respectively, attached to a poly(A) tail of approximately 125 nucleotides. It encodes a preprotein of 166 amino acids, including a prepeptide of 22 amino acids. No overall sequence homology was found between PSP-S and other pancreatic proteins. Some homology with several serine proteases was observed in the COOH-terminal region, however. The mRNA levels of PSP-S, trypsinogen, chymotrypsinogen, and colipase in CCP and control pancreas were compared. PSP-S mRNA was three times lower in CCP than in control, whereas the others were not altered. It was concluded that PSP-S gene expression is specifically reduced in CCP patients.

Topics: Adolescent; Adult; Amino Acid Sequence; Bacteriophage lambda; Base Sequence; Blotting, Northern; Calcium-Binding Proteins; Chronic Disease; Chymotrypsinogen; Colipases; DNA; Female; Gene Expression Regulation; Humans; Lithostathine; Male; Molecular Sequence Data; Nerve Tissue Proteins; Nucleic Acid Hybridization; Pancreatic Juice; Pancreatitis; RNA, Messenger; Sequence Homology, Nucleic Acid; Trypsinogen

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.

Topics: Acute Disease; Amino Acid Sequence; Anions; Cations; Chromatography, Affinity; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Kidney Diseases; Molecular Sequence Data; Molecular Weight; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

Immunoreactive trypsin in serum can be divided into trypsinogen and trypsin-alpha 1-proteinase inhibitor (alpha 1PI) complexes. These were studied separately in serum from 204 patients with acute gastro-intestinal symptoms. Elevated levels of both trypsinogen and trypsin-alpha 1PI complexes were seen in patients with acute pancreatitis. Elevated levels of trypsinogen and normal or slightly elevated levels of trypsin-alpha 1PI complexes were seen in patients with biliary tract diseases. An isolated increase in the concentration of trypsin-alpha 1PI complexes with normal trypsinogen and amylase levels were seen in patients with perforated ulcer. This third cluster may result from an absorption of active trypsin from the peritoneal cavity. Small amounts of trypsin-alpha 1PI complexes were present also in serum from patients free from pancreatic disease. The results in this study show that high levels of trypsin-alpha 1PI complexes in serum are seen mainly in patients with acute pancreatitis. However, elevated levels are also seen in other pathological conditions in the upper gastrointestinal tract; therefore an assay for these complexes is not a specific diagnostic test for acute pancreatitis.

Topics: Abdomen, Acute; alpha 1-Antitrypsin; Biliary Tract Diseases; Gastritis; Humans; Pancreatic Neoplasms; Pancreatitis; Peptic Ulcer; Trypsinogen

Sixty-one patients (1 to 18 1/2 years of age) with acute pancreatitis were evaluated. In over one third of cases, acute pancreatitis was one feature of a multisystem disease (Reye syndrome, sepsis, shock, hemolytic-uremic syndrome, viral infections). Other common causes included blunt trauma (15%), acquired or congenital structural defects (10%), metabolic diseases (10%), and drug toxicity (3%). In 25% of cases, no cause was identified. All conscious patients complained of abdominal pain, but the location, severity, and duration of pain were extremely variable. Vomiting was a common symptom. Ultrasonography was helpful in establishing the diagnosis and for assessment of complications such as pseudocyst formation. Endoscopic retrograde cholangiopancreatography was used to identify structural or anatomic lesions in patients with recurrent acute pancreatitis. Serum cationic trypsin(ogen) was superior to amylase in the early diagnosis of acute pancreatitis, and was more consistently elevated during the first 5 days in the hospital. Patients were managed conservatively with complete bowel rest, gastric decompression, intravenous fluid therapy, and pain relief. Pancreatic pseudocysts occurred in 10% of patients. There were 13 fatalities, all in patients with a severe multisystem disorder. Recurrences of acute pancreatitis were noted only in certain diagnostic groups: idiopathic pancreatitis, structural anomalies of the pancreaticobiliary tree, metabolic disorders, and (in a single patient) familial pancreatitis.

Topics: Acute Disease; Adolescent; Adult; Amylases; Child; Child, Preschool; Combined Modality Therapy; Humans; Infant; Male; Pancreatitis; Radioimmunoassay; Retrospective Studies; Trypsinogen

Topics: Alcoholism; Amylases; Animals; Atrophy; Cholecystokinin; Dietary Fats; Male; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Receptors, Cholecystokinin; Trypsinogen

Acute pancreatitis (AP) is believed to result from intraparenchymal activation of trypsin and other digestive enzymes within the pancreas followed by autodigestion of the gland. Gabexate mesilate (FOY), a synthetic guanidino acid ester exhibiting potent and versatile inhibitory actions on a number of proteinases (e.g., trypsin, kallikrein, C1-r, C1 esterase, plasmin, thrombin, phospholipase A2), was examined for its ability to protect the rat pancreas against development of AP induced by pharmacological doses of ceruletide (CRT). Rats were i.v. infused for 6 h with either CRT (5 micrograms/kg/h) or CRT + FOY (50 mg/kg/h). In FOY-treated rats the serum amylase and trypsinogen concentrations were reduced by 60 and 80%, respectively, compared to rats infused with CRT alone. Histologically, the extent of acinar cell vacuolization in the pancreas was significantly reduced and interstitial edema, although not assessed by quantitative morphometric techniques, appeared to be qualitatively lessened in the FOY-treated rats. The ability of FOY to inhibit significantly AP produced by supramaximal doses of CRT, coupled with its inhibitory properties on components of the coagulation and complement cascades, stress the importance of continued research on this compound as a potential therapeutic agent for treatment of AP and its systemic sequelae.

Topics: Acute Disease; Amylases; Animals; Ceruletide; Disease Models, Animal; Gabexate; Guanidines; Infusions, Intravenous; Male; Pancreatitis; Protease Inhibitors; Rats; Rats, Inbred Strains; Trypsinogen

The plasma levels of certain digestive enzymes and protease inhibitors were determined in 40 patients with severe acute pancreatitis diagnosed as gallstone-induced (GP), alcoholic (AP), or idiopathic (IP). On admission, plasma levels of amylase and immunoreactive cationic trypsin(ogen) (IRCT) and elastase 2 (IRE 2) were found to be 50 +/- 10 ng/ml, 340 +/- 64 ng/ml, and 190 +/- 15 ng/ml, respectively, in all groups of patients. These enzymes levels remained high for the first 2 days following hospitalization and then decreased, although amylase and IRCT levels remained elevated above normal values throughout the study period (2 weeks). In general amylase and IRCT were lower in patients with concomitant ascites, pancreatic pseudocysts, or abscesses, and higher in patients who died, as compared to the pancreatitis group as a whole. In these patients, levels of immunoreactive alpha 1-protease inhibitor (alpha 1-PI) and alpha 2-macroglobulin (alpha 2-M) remained relatively constant at the lower end of the normal range throughout the study period. Inhibitor levels in plasma were unaffected by the etiology of pancreatitis or the occurrence of complications. A similar trend was seen with plasma levels of lysosomal hydrolases. The results indicate that in this group of patients, plasma levels of pancreatic digestive enzymes were reflective of the clinical features and severity of the disease, as well as the time following the acute attack that brought the patient to the hospital.

Topics: Acute Disease; Adult; Aged; alpha 1-Antitrypsin; Amylases; Blood Proteins; Female; Humans; Male; Middle Aged; Pancreatic Elastase; Pancreatitis; Protease Inhibitors; Trypsinogen

The variations of serum levels of amylase, pancreatic isoamylase, lipase, trypsinogen, and elastase 1 were evaluated in 21 patients with acute pancreatitis. The patients were studied for a mean period of 7 consecutive days (range 5-12 days) after admission to the hospital. On the day of onset of acute pancreatitis, all enzyme levels were abnormally high; pancreatic isoamylase showed the greatest increase compared with its upper normal limit, whereas the increase increment for elastase 1 was the lowest. Subsequently, all enzyme levels except elastase 1 decreased in a parallel fashion. On the eighth day of the study only elastase 1 levels were above normal values in all patients examined, while abnormally high values of lipase were found in 85% of the patients, trypsinogen in 58% of the patients, pancreatic isoamylase in 43%, and total amylase in 23%. These results indicate that, for the early diagnosis of acute pancreatitis, the determination of any of these enzymes is equally efficient, but that elastase 1 is the most sensitive marker of acute pancreatic damage in later stages of the disease.

Topics: Acute Disease; Adolescent; Adult; Aged; Aged, 80 and over; Amylases; Female; Humans; Isoamylase; Lipase; Male; Middle Aged; Pancreas; Pancreatic Elastase; Pancreatitis; Trypsinogen

Topics: Amylases; Female; Humans; Pancreatitis; Pre-Eclampsia; Pregnancy; Trypsinogen

In order to investigate the role of circulating free trypsinogen and renal tubular dysfunction in affecting trypsin plasma-urine transfer, serum immunoreactive trypsin (IRT), its urinary output, IRT molecular size distribution, filtrable immunoreactive trypsin, gamma-glutamyltransferase and alpha-glucosidase outputs were studied in 6 control subjects, 9 patients with pancreatic cancer and 15 with chronic pancreatitis. The majority of immunoreactivity was always eluted at a molecular weight of about 24,000 and might therefore be considered as free trypsinogen. Variable amounts of IRT at higher molecular weights, possibly represented by trypsin-inhibitor complexes, were also detected. Increasing IRT levels were generally accounted for by free trypsinogen, regardless of the nature of the disease. Unlike serum free trypsinogen levels, renal tubular damage, evaluated by means of the excretion of two high-molecular weight urinary enzymes, seems to play a prominent role in explaining trypsin plasma-urine transfer.

Topics: Adolescent; Adult; Aged; alpha-Glucosidases; Carcinoma, Intraductal, Noninfiltrating; Chronic Disease; Female; gamma-Glutamyltransferase; Humans; Kidney Diseases; Kidney Tubules; Male; Metabolic Clearance Rate; Middle Aged; Molecular Weight; Pancreatic Neoplasms; Pancreatitis; Trypsin; Trypsinogen

Ultrasonic monitoring of the pancreas following secretin stimulation has shown to cause a marked dilatation of Wirsung duct; whether this phenomenon is due to the stimulation of pancreatic secretion and/or to the effect of secretin on the sphincter of Oddi (SO) motility is unknown. In the present study pancreatic scan after secretin was performed in 11 patients with nonpancreatic diseases after premedication with glucagon (inhibition of both pancreatic secretion and SO motility) or tyropramide (inhibition of SO motor function) and in patients with different degrees of pancreatic insufficiency. Serum immunoreactive trypsinogen (IRT) levels were measured in all the subjects during the test. Premedication with glucagon completely abolished both Wirsung enlargement and serum IRT increase, while tyropramide significantly reduced, but did not abolish, the response to secretin. These results suggest that both stimulation of pancreatic secretion and the increase of SO pressure are prerequisites for a full-blown occurrence of the secretin-induced modifications of Wirsung. Within chronic pancreatitis patients, the response to secretin was exaggerated in those with a still preserved pancreatic function and it was lacking in those with severe pancreatic insufficiency.

Topics: Chronic Disease; Common Bile Duct Diseases; Dilatation, Pathologic; Humans; Pancreas; Pancreatic Diseases; Pancreatic Ducts; Pancreatitis; Secretin; Sphincter of Oddi; Trypsinogen; Ultrasonography

We evaluated the behavior of serum cationic trypsinogen (SCT), an enzyme of solely pancreatic origin, in 30 patients with chronic pancreatitis and 25 healthy subjects as a control, after secretin and bombesin stimulation. After both the stimulations, serum cationic trypsinogen is unable to distinguish between the healthy control subjects and the patients with chronic pancreatitis. On the other hand, after secretin, the enzyme is able to separate chronic pancreatitis patients with different levels of exocrine function insufficiency. It does so with a greater statistical significance than that obtained by the rapid injection of bombesin and equal to that of trypsin into the duodenal juice during duodenal intubation. For these reasons, as well as the absence of any side-effects, secretin is preferred to bombesin stimulation in the evaluation of the exocrine pancreatic function in patients with chronic pancreatitis.

Topics: Adult; Alcoholism; Bombesin; Cations; Chronic Disease; Female; Humans; Male; Middle Aged; Pancreatitis; Secretin; Trypsinogen

A rat model of taurocholate-induced acute pancreatitis has been employed to investigate the effect of heparin on the protease-antiprotease balance. Heparin was applied intraperitoneally at a dose of 6 mg/kg body weight during 24 hrs. At 24 and 48 hours of acute pancreatitis, heparin evidently diminished the consumption of trypsinogen in pancreatic tissue and decreased trypsin generation. The use of heparin prevented the consumption of alpha 1 anti-chymotrypsin, alpha 1-anti-trypsin and AT-III in pancreatic tissue, whereas in plasma the concentration of the mentioned inhibitors was restored or even increased. Heparin does not affect evidently lowered alpha 2-macroglobulin concentration, either in pancreatic tissue or in plasma. We conclude that heparin applied in acute pancreatitis markedly moderates the dysfunction of protease-antiprotease balance both in plasma and in pancreatic tissue.

Topics: Acute Disease; alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Antithrombin III; Heparin; Male; Pancreas; Pancreatitis; Peptide Hydrolases; Protease Inhibitors; Rats; Trypsin; Trypsinogen

Activation of trypsinogen in acute pancreatitis results in subsequent increases in plasma levels of trypsin bound to the inhibitors alpha 1-protease inhibitor (alpha 1-PI) and alpha-macroglobulin (alpha-M). It seems logical to speculate that plasma levels of these inhibitor-bound forms of trypsin may reflect the degree of intrapancreatic zymogen activation and that determination of such parameters may be of diagnostic and prognostic value. In order to test this hypothesis, the concentrations of trypsinogen and of trypsin bound to alpha 1-PI have been determined in serial plasma samples from rats who died (N = 7) and survived (N = 5) following induction of pancreatitis with taurocholate. Since the other major reaction product of active trypsin in plasma, alpha-macroglobulin-bound trypsin, cannot be measured directly, the plasma levels of trypsin-like amidase activity were determined to estimate the concentration of trypsin-alpha-M complex. Shortly after induction of pancreatitis, elevated levels of trypsinogen were present in plasma, but no alpha 1-PI-bound trypsin could be detected. Trypsin-alpha 1-PI complex continuously increased over the time course of pancreatitis in animals that died. In contrast, the plasma levels of trypsin-alpha 1-PI complex were lower in animals that survived, peaked around 15 hr postinduction at levels (182 +/- 53 ng/ml) significantly lower than those in dying animals (543 +/- 346 ng/ml), and fell during the following 48 hr. There was a significant correlation between plasma trypsin-like amidase activity and plasma alpha 1-PI-bound trypsin. Our data demonstrate that the concentration of activated forms of plasma trypsin in the bloodstream are correlated with mortality in experimental pancreatitis.

Topics: alpha 1-Antitrypsin; Animals; Enzyme Activation; Pancreatitis; Protein Binding; Rats; Rats, Inbred Strains; Trypsin; Trypsinogen

The currently available radioimmunoassays of trypsin measure total immunoreactive trypsin (EC 3.1.1.7), which includes both trypsinogen and alpha 1-protease inhibitor-bound trypsin. Hitherto, the only way to differentiate these two forms of trypsin has been to fractionate them on a gel-filtration column. We describe here a solid-phase immunoenzymometric assay that rapidly measures the amount of cationic trypsin bound to alpha 1-protease inhibitor in the plasma of rats with experimental pancreatitis. The assay specifically measures this complex within the range from 0.2 to 5.0 ng without interference by high concentrations of free alpha 1-protease inhibitor. The high correlation (r = 0.985) of the values obtained by size fractionation and by this assay demonstrates the accuracy of the assay, which is the first single-tube method for determining this form of activated cationic trypsin in plasma.

Topics: alpha 1-Antitrypsin; Animals; Blood Proteins; Immunoenzyme Techniques; Macromolecular Substances; Pancreatitis; Radioimmunoassay; Rats; Trypsin; Trypsinogen

Topics: Acute Disease; Animals; Ceruletide; Choline Deficiency; Disease Models, Animal; Duodenum; Enzymes; Ethionine; Humans; Lysosomes; Pancreas; Pancreatic Ducts; Pancreatitis; Trypsin; Trypsinogen

Topics: Acute Disease; Adult; Aged; Clinical Enzyme Tests; Female; Glycoside Hydrolases; Humans; Isoamylase; Male; Middle Aged; Pancreatitis; Trypsinogen

The levels of serum immunoreactive trypsinogen and P-isoamylase in response to Bombesin intravenous infusion were evaluated in 25 controls, 18 patients with documented chronic pancreatitis, and nine subjects with nonpancreatic gastroenterological diseases. Mean immunoreactive trypsinogen peak values were significantly higher in controls and gastroenterological diseases than in chronic pancreatitis, but there was marked overlap in individual values between the three groups. As for P-isoamylase, a statistical difference was detected only between mean peak concentrations of control versus chronic pancreatitis. Integrated responses for both enzymes did not result in a better discrimination between controls, chronic pancreatitis, and gastroenterological diseases. This study confirms that evocative tests are of limited value in the diagnosis of pancreatic disease.

Topics: Bombesin; Chronic Disease; Female; Glycoside Hydrolases; Humans; Isoamylase; Male; Pancreas; Pancreatic Function Tests; Pancreatitis; Trypsinogen

Topics: Acetaldehyde; Acute Disease; Amylases; Animals; Ethanol; Female; In Vitro Techniques; Mice; Pancreas; Pancreatitis; Proteins; Trypsinogen

The sensitivity and specificity of five assays used to diagnose acute pancreatitis were studied: two amylase assays; one lipase; one trypsinogen; and one pancreatic isoamylase. Thirty-nine patients with acute pancreatitis were compared to 127 controls with abdominal pain. Using the upper limit of normal both amylase assays appeared sensitive but somewhat nonspecific (specificities of 88.9% and 86%, respectively). The trypsinogen and pancreatic isoamylase assays were also relatively nonspecific (specificity of 82.8% and 85.1%). Most nonspecific elevations occurred between a one- and twofold elevation of each assay. Lipase, however, maintained excellent specificity (99%) at its upper limit of normal. If the level of best cutoff is used instead (the level that best enhances sensitivity and specificity), the specificities of both amylase assays, as well as the trypsinogen and pancreatic isoamylase assays, exceed 95%. At the best cutoff level, trypsinogen maintains a qualitative advantage in sensitivity over lipase or pancreatic isoamylase (97.4% as compared to 86.5% and 84.6%).

Topics: Acute Disease; Amylases; Clinical Enzyme Tests; Evaluation Studies as Topic; Humans; Isoamylase; Lipase; Pancreas; Pancreatitis; Radioimmunoassay; Trypsinogen

We employed a radioimmunoassay for human cationic trypsin to define the time course for alterations in the molecular forms of this enzyme in plasma from patients with pancreatitis. Six patients developed acute pancreatitis as a complication of a known disorder [three, Reye's syndrome; two, hemolytic uremic syndrome (HUS); one, choledochal cyst]. The immunoreactive forms of cationic trypsin were determined by gel filtration of each plasma sample followed by radioimmunoassay of the column fractions. Early in the course of the disease, predominantly free trypsinogen was released into the circulation in five patients. In the three patients with Reye's syndrome, subsequent plasma samples showed, in addition to free trypsinogen, increasing amounts of immunoreactive trypsin complexed to alpha 2-macroglobulin and alpha 1-protease inhibitor. In contrast, subsequent samples from the two patients with HUS contained little or no inhibitor-bound trypsin. The remaining patient had intermediate concentrations of cationic trypsin complexed to these two circulating protease inhibitors. Five patients died and postmortem studies showed a striking correlation between the histological severity of acute pancreatic inflammation and the amount of immunoreactive trypsin complexed to alpha 2-macroglobulin and alpha 1-protease inhibitor. This preliminary study suggests that measurement of alpha 2-macroglobulin or alpha 1-protease inhibitor-bound trypsin may be a useful method of monitoring the progression and severity of disease in patients with acute pancreatitis. Characterization of serial changes in the forms of circulating pancreatic proteases may enhance our understanding of time-dependent pathophysiologic events, possibly leading to improved forms of specific therapy.

Topics: Acute Disease; alpha 1-Antitrypsin; alpha-Macroglobulins; Amylases; Blood Proteins; Child; Chromatography, Gel; Female; Hemolytic-Uremic Syndrome; Humans; Male; Pancreatitis; Protease Inhibitors; Radioimmunoassay; Reye Syndrome; Time Factors; Trypsin; Trypsinogen

The cascade enterokinase-trypsinogen-prophospholipase A2 lecithin, generating trypsin, phospholipase A2 and lysolecithin, respectively, was studied in vitro using a novel phospholipase A2 assay. The rate of enterokinase catalysed activation of trypsinogen was maximal at 4 mmol/1 glycodeoxycholic acid; higher concentrations of bile salt progressively inhibited enterokinase activity. Net phospholipase A2 activity in reaction mixtures was critically dependent on the trypsin/prophospholipase A2 molar ratio. Lecithin hydrolysis by phospholipase A2 was dependent on the bile salt/lecithin molar ratio and was optimal at 1.25 to 1. The addition of enterokinase to lecithin and bile salt mixtures, containing trypsinogen and prophospholipase A2 at presumed pathophysiological concentrations, resulted in the generation of concentrations of lysolecithin lytic for pancreatic acinar cells within 5 min. These findings would support the concept that the entry of bile containing active enterokinase into the pancreatic duct system in vivo may in some cases be involved in the initiation of necrotising acute pancreatitis in man.

Topics: Acute Disease; Endopeptidases; Enteropeptidase; Enzyme Activation; Enzyme Precursors; Glycodeoxycholic Acid; Humans; Hydrolysis; Kinetics; Lysophosphatidylcholines; Necrosis; Pancreatitis; Phospholipases; Phospholipases A; Phospholipases A2; Trypsin; Trypsinogen

Previous studies, in selected populations, have determined that a low serum trypsinogen can be seen in chronic exocrine pancreatic disorders (CP) and primary diabetes mellitus (DM). In this study, we investigated the predictive value of a low serum trypsinogen. The study population consisted of 488 consecutive emergency room patients admitted to our hospital on whom a serum amylase was drawn by the emergency room staff. Of the sera drawn, 418 were saved and tested for immunoassayable trypsinogen. Ten of 418 (2.4%), had a low level of this marker (less than 10 ng/ml). Of these 10, four had obvious historical or clinical evidence of CP during their initial hospitalization. Six patients, however, had no initial evidence of CP. Follow-up was obtained in three of the six, and all three had evidence of CP despite absence of symptoms. Of the 418 patients, 37 had DM. A low trypsinogen was found in three of these 37, and all three had concomitant CP. We conclude that this new assay has excellent predictive value in diagnosing chronic exocrine pancreatic disorders.

Topics: Adult; Aged; Chronic Disease; Diabetes Mellitus; Emergency Service, Hospital; Female; Humans; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

Topics: Chronic Disease; Clinical Enzyme Tests; Humans; Pancreatitis; Trypsinogen

The levels of amylase, trypsinogen, and trypsin-alpha 1-protease inhibitor complexes, both in serum and in peritoneal fluid, were correlated to the severity and clinical course in 27 attacks of acute pancreatitis. Serum levels of trypsin-alpha 1-protease inhibitor complexes on admission correlated well with the severity and clinical course of the disease, whereas serum levels of amylase and trypsinogen did not. This may be of clinical importance in differentiating severe acute pancreatitis from milder and self-limiting forms of the disease.

Topics: Acute Disease; alpha 1-Antitrypsin; Amylases; Ascitic Fluid; Humans; Pancreatitis; Trypsinogen

The protective role of alpha 2-macroglobulin, alpha 1-antitrypsin and Aprotinin against trypsin-induced effects on C3 and kininogen was studied in a human in vitro model. When human cationic trypsin was added to human serum or plasma, there was a gradual saturation of alpha 2-macroglobulin and later of alpha 1-antitrypsin. When alpha 2-macroglobulin was 70% saturated, there was a prompt cleavage of both C3 and kininogen, in spite of 80% free and active alpha 1-antitrypsin. These biochemical changes and antiprotease levels are identical to our findings in patients with acute pancreatitis, especially in their peritoneal exudate. Very high concentrations of Aprotinin, 5-15 times higher than ever used clinically, blocked the cleavage of both C3 and kininogen, while doses commonly used clinically were without significant effect. The clinical implications are: A trypsin-induced activation of both the complement and kinin system with clinical consequence is possible in patients with acute pancreatitis because of very low alpha 2-macroglobulin levels. Aprotinin in adequate doses, 5-15 times higher than ever used clinically, seems to protect against activation of two systems.

Topics: Acute Disease; alpha 1-Antitrypsin; alpha-Macroglobulins; Blood Proteins; Complement C3; Humans; Immunoelectrophoresis, Two-Dimensional; Kinetics; Pancreatic Elastase; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

In short-term experiments (25 or 72 h) oral trypsin inhibitor administration to pancreatitic rats significantly decreased survival rate, whereas oral trypsin administration had no effect in this respect. Neither treatment influenced the activities of amylase in serum, pancreatic tissue or ascites. Trypsin given in excess together with the trypsin inhibitor abolished the deleterious effects on survival caused by the trypsin inhibitor. In a long-term experiment in healthy rats oral trypsin inhibitor ingestion caused a significant increase in pancreatic wet weight, protein concentration and activities of amylase, lipase and trypsinogen in pancreatic tissue; again, trypsin administration had no effect. The data support the idea that oral trypsin inhibitor administration causes release of cholecystokinin (CCK) or CCK-like factors from the intestine by interfering with the negative feedback regulation exerted by intraluminal trypsin. The results of the short-term experiments further indirectly suggest that even small amounts of trypsin within the intestine - as in acute pancreatitis - can exert the feedback regulation. Finally, the results of the long-term experiment suggest that oral administration of trypsin does not exert any suppressive effects on pancreatic wet weight and pancreatic enzyme content.

Topics: Acute Disease; Amylases; Animals; Aprotinin; Feedback; Lipase; Male; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Trypsin; Trypsinogen

A double antibody solid phase radioimmunoassay for the determination of cationic trypsin in complex with alpha 1-antitrypsin in human sera is described. No immunoreactive trypsin in complex with alpha 1-antitrypsin can be detected in normal sera. In sera from 8 patients with acute pancreatitis levels between 100 and 1000 micrograms/l are seen. The amount of trypsin in complex with alpha 1-antitrypsin in serum in acute pancreatitis equals or exceeds the amount of trypsinogen. Serum levels of trypsin in complex with alpha 1-antitrypsin also remain elevated longer than the trypsinogen levels in acute pancreatitis.

Topics: Acute Disease; alpha 1-Antitrypsin; Chromatography, Gel; Humans; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Enterokinase activates trypsinogen very rapidly and is itself resistant to proteolysis and endogenous inhibitors in blood and pancreas. Using a novel one-stage specific catalytic assay capable of detecting enterokinase in the presence of trypsin inhibitors, we have positively identified catalytically active enterokinase in human bile in each of 14 patients studied. Since the presence of active enterokinase in human bile was not explicable by duodeno-biliary reflux, enterokinase must have followed the pathway from gut to blood to liver to bile, previously identified in greater detail experimentally. We suggest that entry into the pancreatic duct system of bile-borne active enterokinase from the common bile duct may trigger necrotising acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Bile; Chromatography, Gel; Endopeptidases; Enteropeptidase; Female; Humans; Male; Methods; Middle Aged; Molecular Weight; Pancreatitis; Time Factors; Trypsinogen

A new radioimmunoassay to serum trypsinogen (Cis Trypsik) was tested in several patient populations. A low serum trypsinogen level (less than 10 ng/ml) was found in 69.2% of 13 patients with chronic pancreatic insufficiency (CPI), in 100% of 10 patients with 95-100% pancreatectomy but only in 14% of 14 patients with cancer of the pancreas. A low trypsinogen level was not found in any of 68 control subjects or 10 patients with nonpancreatic steatorrhea. Nine patients with CPI or 95% pancreatectomy were retested a mean of six months after initial testing. Four of these nine (44.4%) had a significant variation in serum trypsinogen which would have led to a different diagnostic interpretation (two went from low to normal levels and two from normal to low levels). A mixed meal had little effect on serum trypsinogen levels in five of six patients with CPI, and pancreatic enzyme replacement therapy had no consistent effect on the serum trypsinogen level in seven patients with CPI or 95% pancreatectomy. It is speculated that minor subclinical episodes of focal pancreatitis may effect the serum trypsinogen level. Although there can be considerable variability using this assay, it still offers important clinical utility. A low trypsinogen level points to a chronic pancreatic process with excellent specificity. A normal trypsinogen level is of no help and should be repeated if clinical suspicion of chronic pancreatitis remains high.

Topics: Chronic Disease; Female; Humans; Male; Pancreatitis; Trypsinogen

SDS electrophoresis on polyacrylamide gels of purified trypsinogen 1 has shown the occurrence of a proteolysis in some molecules during long storage at -20 degrees C. This proteolyzed trypsinogen gives a positive reaction with an antiserum directed against the precipitate protein, major protein of about 14 000 molecular weight extracted from precipitates present in the pancreatic juice of patients with chronic pancreatitis. The autoactivation of proteolyzed trypsinogen 1 liberates a polypeptide of 14 000 molecular weight which is immunologically identical to the precipitate protein. These results show that the major protein present in pancreatic precipitates (and pancreatic stones) of patients with chronic pancreatitis is a degradation product of trypsinogen 1 liberated by a proteolysis which necessarily requires a premature zymogen activation in the disease.

Topics: Chronic Disease; Electrophoresis, Polyacrylamide Gel; Humans; Immunoelectrophoresis; Isoenzymes; Molecular Weight; Pancreatic Juice; Pancreatitis; Trypsinogen

Topics: Celiac Disease; Clinical Enzyme Tests; Exocrine Pancreatic Insufficiency; Humans; Pancreatitis; Radioimmunoassay; Reagent Kits, Diagnostic; Trypsin; Trypsinogen

Topics: Chronic Disease; Humans; Pancreatic Juice; Pancreatitis; Trypsinogen

Unconscious rats given intravenous ceruletide (diethylamine salt of the decapeptide caerulein) in large pharmacologic doses consistently developed moderate acute pancreatitis by 3 h and florid pancreatitis by 6 h. Biochemical serum markers of acute pancreatitis tended to parallel the severity of the pancreatic damage. In 50% of the rats, mesenteric fat necrosis was present, free peritoneal fluid containing massive elevations of trypsinogen and amylase were noted in most animals. Intravenous secretion at a low dose given simultaneously with ceruletide exerted a variable protective effect on the pathological process. A high dose of secretin produced a striking macroscopic, microscopic, and biochemical protective effect on ceruletide-induced pancreatitis. High resolution light microscopy and electron microscopy showed a marked cellular disorganization in the acini of animals treated with ceruletide alone. By contrast, there was a striking apical redirection of zymogen granules in acini of the animals treated with secretin. The results of this study suggest that high dose intravenous secretin may exert a beneficial effect on acute pancreatitis.

Topics: Acute Disease; Amylases; Animals; Ceruletide; Edema; Fat Necrosis; Lipase; Male; Pancreas; Pancreatitis; Rats; Rats, Inbred Strains; Secretin; Trypsinogen

Immunoreactive trypsin was localized with the peroxidase-antiperoxidase technique in normal human pancreatic tissue and in the glands of patients suffering from acute or chronic pancreatitis. In the normal pancreas and in the histologically normal areas of the inflamed pancreas, trypsin was detected in the zymogen granules of acinar cells and in ductal secretory material. During acute pancreatitis, three characteristic changes were observed: (1) separate acinar cell fragments in early lesions; (2) decreased and evenly dispersed staining in necrotic acinar cells, and (3) intensive reaction in plugs in acinar lumina in advanced lesions. In chronic pancreatitis, the localization of trypsin in acinar cells was similar to that in normal pancreas. Some proteinaceous plugs in dilated pancreatic ducts were weakly immunoreactive. The results show that the tissue distribution of immunoreactive trypsin is altered in acute pancreatitis.

Topics: Acute Disease; Adult; Aged; Chronic Disease; Cytoplasmic Granules; Enzyme Precursors; Female; Humans; Immunoenzyme Techniques; Male; Middle Aged; Pancreas; Pancreatitis; Trypsin; Trypsinogen

Topics: Animals; Humans; Immunoenzyme Techniques; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Topics: Acute Disease; Adult; Animals; Cacodylic Acid; Callitrichinae; Cattle; Celiac Disease; Child; Chlorocebus aethiops; Chronic Disease; Dianisidine; Duodenum; Endopeptidases; Enteropeptidase; Guinea Pigs; Histocytochemistry; Humans; Intestine, Small; Mice; Mice, Inbred Strains; Oligopeptides; Pancreatitis; Rats; Swine; Swine, Miniature; Trypsinogen

This study was performed to determine the effects of exogenous prostaglandin and a prostaglandin synthetase inhibitor on experimental pancreatitis in mice. An ethionine-supplemented choline-deficient diet was used to induce pancreatitis in 4-6-wk-old Swiss Webster mice. Mice were injected subcutaneously with 16,16-dimethyl prostaglandin E2 (0.1, 1.0, 10 micrograms X kg-1 X day-1), indomethacin (0.05, 0.5, 5 mg X kg-1 X day-1), or saline for 7 days. The ethionine-supplemented choline-deficient diet was introduced 24 h after the first injection, and animals ate the test diet for 48 h. A 55% mortality was observed in control animals (n = 100) treated with carrier alone. Treatment with 10 micrograms X kg-1 X day-1 of 16,16-dimethyl prostaglandin E2 significantly decreased (p less than 0.01) mortality to 12% (n = 100). Improved survival was accompanied by a significant (p less than 0.05) decrease in the pancreatic content of free chymotrypsin and a decrease in histologic damage. Treatment with 5 mg X kg-1 X day-1 of indomethacin (n = 30) significantly (p less than 0.01) increased mortality in diet-treated rats from a control rate of 55% to 100%. These studies demonstrate a protective effect of prostaglandin on the pancreas and suggest a role for endogenous prostaglandins in the pathophysiology of pancreatitis.

Topics: 16,16-Dimethylprostaglandin E2; Acute Disease; Animals; Chymotrypsin; Diet; Female; Indomethacin; Mice; Pancreas; Pancreatitis; Prostaglandins E, Synthetic; Trypsin; Trypsinogen

The aim of the present study was to determine the effect of prolonged ethanol intake on the morphology and protein metabolism in the rat pancreatic acinar cells. Weight-matched triplets of Sprague-Dawley rats were fed Lieber-DeCarli diet containing 5% (wt/vol) concentration of ethanol, isocaloric amounts of Lieber-DeCarli diet, or rat chow ad libitum for 6, 12, and 18 mo. In the ethanol-fed group, histologic studies by light microscopy showed absence of protein plugs in the pancreatic ducts and/or pancreatitis, but electron-microscopic evaluation revealed progressive accumulation of lipid droplets in acinar and ductal cells. No definite changes in the mitochondria and endoplasmic reticulum were noticed. Biochemical studies revealed increased specific activity of trypsinogen, chymotrypsinogen, and lipase, and decreased specific activity of amylase. Trypsin-inhibiting capacity was decreased in the tissue and in the medium in a progressive fashion. There was no increase in the secretion of total protein. These data show a complex and a nonparallel alteration of specific digestive enzymes and trypsin inhibitor in this model of chronic ethanol intoxication that may be of relevance to occurrence of pancreatitis.

Topics: Alcoholism; Amylases; Animals; Chymotrypsinogen; Disease Models, Animal; Ethanol; Humans; Lipase; Male; Pancreas; Pancreatitis; Proteins; Rats; Rats, Inbred Strains; Trypsin Inhibitors; Trypsinogen

Acute hemorrhagic pancreatic necrosis (AHPN) is induced in young female mice fed fo 4 days a choline-deficient diet containing diet 0.5% DL-ethionine (CDE). Contrary to females, male mice do not develop AHPN when fed the same diet. For determination of whether estrogens are involved in the induction of AHPN, estradiol-treated male mice were fed the CDE diet. In such estrogen-treated male mice, the mortality rate, incidence of AHPN, and alterations in biochemical parameters of the pancreas and of serum were similar to those induced by the CDE diet in females.

Topics: Acute Disease; Amylases; Animals; Blood Proteins; Cathepsin B; Cathepsins; Choline Deficiency; Diet; Estradiol; Female; Male; Mice; Necrosis; Pancreas; Pancreatitis; Trypsin; Trypsinogen

Topics: Adult; Aged; Female; Gallstones; Hepatitis, Alcoholic; Humans; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Serum immunoreactive trypsinogen (IT) levels were measured in 479 normal controls and in 604 patients (510 with nonpancreatic diseases and 94 with pancreatic diseases) in order to evaluate the distribution of IT values in the control population and the accuracy of the assay in the diagnosis of pancreatic diseases. It concentrations were normally distributed in the healthy population; children showed mean IT values significantly lower than adults. The sensitivity, specificity, and predictive value of a positive and negative result in diagnosing acute or chronic pancreatitis were evaluated vs normals and vs normals plus all patients. With an IT value higher than 80 ng/ml, the likelihood that a patient is not affected by acute pancreatitis is less than 5%. An IT value lower than 9 ng/ml detected 44% of chronic pancreatitis and was related to a 52% probability of such a condition. The 48% false positive results also include patients with pancreatic tumor (31% of cases), so that the chance of finding reduced IT levels in subjects without pancreatic damage drops to 17%. In view of the low prevalence of pancreatic diseases, IT assay should not be taken into consideration as a diagnostic screening test in the general population and its use should be limited to a hospitalized population.

Topics: Adolescent; Adult; Aged; Cations; Child; Child, Preschool; False Positive Reactions; Female; Humans; Infant; Male; Middle Aged; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Radioimmunoassay; Trypsinogen

Topics: alpha 1-Antitrypsin; alpha-Macroglobulins; Carrier Proteins; Cell-Free System; Humans; Hypocalcemia; Pancreatitis; Parathyroid Hormone; Trypsin; Trypsinogen

A canine model of bile-induced pancreatitis has been employed to investigate time-dependent changes in the molecular forms of trypsin in blood and ascitic fluid in this disease. The distribution of immunoreactive trypsin as trypsinogen and trypsin bound to plasma inhibitors in ascitic fluid and plasma during the course of the disease has been investigated by means of a radioimmunoassay for canine pancreatic cationic trypsin. In addition, trypsinlike amidase activity was determined in plasma and ascitic fluid using Z-Gly-Gly-Arg-beta-Nap as substrate. Early plasma and ascitic fluid samples in four dogs that died contained primarily trypsinogen, while extensive activation of trypsinogen to alpha 2-macroglobulin and alpha 1-protease inhibitor-bound trypsin occurred in the course of the disease. A fifth dog survived and showed little activation of trypsinogen. In the four dogs that died, the levels of trypsinlike amidase activity in the ascitic fluid were substantial throughout the course of the disease. The plasma levels of trypsinlike activity in these animals were much lower, but increased during the disease process. The dog that survived had lower concentrations of trypsinlike activity in ascitic fluid and plasma. These results suggest that activation of trypsinogen resulting in inhibitor-bound forms of trypsin in ascitic fluid and plasma is important in the pathogenesis of acute pancreatitis.

Topics: alpha-Macroglobulins; Amidohydrolases; Animals; Ascitic Fluid; Bile; Cations; Chromatography, Gel; Dogs; Female; Pancreatitis; Protease Inhibitors; Radioimmunoassay; Time Factors; Trypsin; Trypsinogen

Prostaglandins have been noted to have a "protective" effect against gastrointestinal mucosal injury induced by a wide variety of agents although possible protective effects of prostaglandins on injury to other tissues have not been reported. We have tested the effect of prostaglandin E2 (PGE2) on acute experimental pancreatitis induced by feeding young female mice a choline-deficient ethionine-supplemented (CDE) diet for 24 hr. Administration of 0.05--0.20 microgram PGE2/g body wt 1 hr before and 4 hr after institution of the CDE diet lowered the mortality rate of diet-induced pancreatitis from 56% to 31%. Larger and smaller doses of PGE2 were without effect. Administration of PGE2 (0.10 microgram/g body weight) diminished the rise in in-vitro LDH discharge and the increase in "free" Cathepsin D activity which occur during diet-induced pancreatitis. Similarly, PGE2 (0.10 microgram/g body wt) diminished the magnitude of the increase in in-vitro protein discharge and the elevated concentrations of trypsinogen and chymotrypsinogen in pancreas fragments taken from mice given the CDE diet. These findings indicate the PGE2 has a protective effect against CDE diet-induced acute experimental pancreatitis. The Cathespin D and LDH changes noted during CDE diet-induced pancreatitis suggest that this diet may decrease membrane integrity and thus allow these enzymes to leak out of the lysosomes and acinar cell, respectively, during pancreatitis. Although the basis for the protective effect of PGE2 remains unclear, our observations suggest that the prostaglandin may act to reduce the alteration in membrane integrity which occurs during CDE-diet induced pancreatitis.

Topics: Amylases; Animals; Cathepsins; Choline Deficiency; Chymotrypsinogen; Diet; Ethionine; Female; In Vitro Techniques; L-Lactate Dehydrogenase; Mice; Pancreatitis; Prostaglandins E; Proteins; Trypsinogen

Radioimmunoassays for anionic and cationic dog trypsins are described. Characterization of the immunoreactivities in sera from fasting dogs demonstrated the presence of the two proenzymes only. Fasting sera from 10 dogs contained anionic and cationic trypsinogen in concentrations between 17-110 micrograms/l and 7-19 micrograms/l, respectively. Induction of experimental pancreatitis in dogs was accompanied by a large increase of immunoreactive anionic and cationic trypsins in the circulation. During the progress of the pancreatitis, immunoreactive trypsin with larger molecular weight than trypsinogen appeared. This high molecular weight immunoreactive trypsin was not seen in serum after intravenous injection of pancreatic juice in dogs. The high molecular weight immunoreactive trypsin probably consists of trypsin in complex with protease inhibitors. In vitro studies showed that the immunoreactivity of trypsin decreased considerably after binding to alpha 1-antitrypsin or alpha-macroglobulins.

Topics: alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Bile; Dogs; Molecular Weight; Pancreatic Juice; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

In acute sodium-taurocholate-induced pancreatitis in the rat, peritoneal dialysis reduced serum amylase levels and the amount of fat necrosis, but did not influence the damage to the pancreas itself. Pancreatic ascites obtained in the early course of the disease was found to have a hypotensive effect when given intraperitoneally to healthy rats. This effect vanished in the later course of acute experimental pancreatitis and was reduced by acidification of the ascites or by administration of an antihistaminic drug. Thus the beneficial effect of continuous peritoneal dialysis on survival time and mortality rate seems to be of systemic origin.

Topics: Acute Disease; Amylases; Animals; Antazoline; Ascites; Ascitic Fluid; Blood Pressure; Hydrogen-Ion Concentration; Injections, Intraperitoneal; Leukocyte Count; Male; Pancreatitis; Peritoneal Dialysis; Phospholipases; Rats; Taurocholic Acid; Trypsin; Trypsinogen

The molecular forms of immunoreactive pancreatic cationic trypsin in sera of patients with acute pancreatic inflammation have been characterized using a radioimmunoassay technique that is capable of detecting trypsinogen as well as trypsin bound to alpha 1-antitrypsin. Trypsin bound to alpha 2-macroglobulin is not immunoreactive under normal assay conditions. However, alpha 2-macroglobulin-bound trypsin can be detected after gel filtration of serum on Bio-Gel A-0.5 m and acid treatment of column fractions. The average serum level of immunoreactive cationic trypsin from 20 patients with acute pancreatic inflammation was 1,590 ng/ml. An average normal value of 26 ng/ml has been obtained previously. Serum samples from 14 patients with pancreatic inflammation were chromatographed under conditions that resolve trypsinogen, alpha 1-antitrypsin-bound trypsin, and alpha 2-macroglobulin-bound trypsin. In each case, the major portion of the immunoreactive material eluted at a position corresponding to free trypsinogen, while a minor fraction of the immunoreactive material appeared to be trypsin bound to alpha 1-antitrypsin. The zymogen nature of the major peak was confirmed in one case by activation with human enteropeptidase. In 11 of 14 patients, acid treatment of the alpha 2-macroglobulin peak yielded immunoreactive trypsin.

Topics: alpha 1-Antitrypsin; alpha-Macroglobulins; Cations; Enteropeptidase; Enzyme Activation; Humans; Molecular Weight; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

A specific radioimmunoassay has been developed for human pancreatic cationic trypsin. The assay has been employed for the determination of immunoreactive forms of pancreatic cationic trypsin in blood. The trypsin employed as radioiodinated tracer in the assay was inactivated with tosyl-L-lysine chloromethyl ketone (TLCK) to prevent binding of the tracer to the serum inhibitors while maintaining its immunoreactivity. The average normal serum level determined was 26 ng/ml, with a range of 12--41 ng/ml. Eight of nine patients with acute pancreatic inflammation had at least a 15-fold elevation of total serum immunoreactive cationic trypsin. Cationic trypsinogen and cationic trypsin bound to alpha1-antitrypsin cross-react strongly in the radioimmunoassay. Thus it is possible to measure these potential molecular forms of cationic trypsin in serum. When normal human serum was fractionated on Sephadex G-200, all of the immunoreactive material eluted as a single peak of approximately 23,000 mol wt. No cationic trypsin could be detected in association with alpha1-antitrypsin or alpha2-macroglobulin. The 23,000-mol-wt peak was definitively shown to contain trypsinogen by affinity chromatography and by activation with human enteropeptidase. The identification of cationic trypsinogen in blood implies that the zymogen is secreted into the circulation by the pancreas rather than entering the bloodstream via absorption from the intestine.

Topics: Enteropeptidase; Humans; Molecular Weight; Pancreas; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

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.

Topics: Adult; Alcoholism; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Pancreatic Juice; Pancreatic Neoplasms; Pancreatitis; Trypsinogen

The concentration in serum of cathodal trypsinogen has been studied in certain clinical and experimental situations. The concentration correlated with pancreatic amylase activity. Low levels were found in patients with malabsorption due to exocrine pancreatic insufficiency. The concentration rose after endoscopic retrograde cholangiopancreatographic examinations (ERCP). After ERCP, however, no trypsin was detected complexed with protease inhibitors, as is generally found in acute pancreatitis. The trypsinogen concentration in serum also rose in renal failure indicating a renal elimination route for the endogenous trypsinogen.

Topics: Amylases; Endoscopy; Humans; Kidney Failure, Chronic; Malabsorption Syndromes; Pancreas; Pancreatic Diseases; Pancreatic Juice; Pancreatitis; Radiography; Trypsinogen

Studies have been performed on pure pancreatic juice obtained by direct cannulation of the pancreatic duct in 2 patients with acute pancreatitis. The striking abnormalities observed, which were in marked contrast to our observations in 15 normal subjects, were high concentrations of protein throughout the period of secretin stimulation and the sporadic appearance of free proteolytic activity in many 1-min specimens throughout the collection period. In 1 subject repeat studies were performed after resolution of the pancreatitis when the profile observed was normal. Our findings are consistent with the hypothesis that obstruction of ductules and intraductal activation of zymogens may be important in the pathogenesis of acute pancreatitis.

Topics: Acute Disease; Adult; Alcoholism; Cholecystokinin; Chymotrypsinogen; Female; Humans; Male; Pancreatic Juice; Pancreatitis; Proteins; Secretin; Trypsin Inhibitors; Trypsinogen

The levels of the proenzymes trypsinogen and chymotrypsinogen were studied in guinea pigs with pancreatitis induced by injection of sodium taurocholate containing the antibiotic cephalothin. This treatment inhibited the enzyme activities and prolonged the activation times of the proenzymes. Both trypsinogen and chymotrypsinogen content decreased after induction of pancreatitis, but there were no significant changes in the proenzyme contents in relation to injection-to-excision times. Sodium taurocholate and cephalothin were cleared from the pancreas in 2 h. Administration of chlorophyll-a together with the inducer caused a slight increase in proenzyme levels.

Topics: Animals; Cephalothin; Chlorophyll; Chymotrypsinogen; Esterases; Guinea Pigs; Pancreas; Pancreatitis; Taurocholic Acid; Trypsinogen

Topics: Amylases; Child; Cystic Fibrosis; Enteropeptidase; Humans; Infant; Methods; Neutropenia; Pancreas; Pancreatic Cyst; Pancreatic Diseases; Pancreatic Neoplasms; Pancreatitis; Peptide Hydrolases; Syndrome; Trypsinogen

Three aspects of the pathophysiology of acute pancreatitis are discussed: 1. the initiating mechanisms, 2. the mechanisms of the fat necrosis, 3. the processes leading to shock phenomena. It is pointed out that the intraglandular activation of the precursors for both lipolytic and proteolytic enzymes seems to be essential for the initiating mechanisms of the disease. The role of the hormone dependent lipolytic enzyme of the fat tissue is discussed in relation to the occurrence of extrapancreatic fat necrosis. The role of the vaso-active compounds, such as plasma kinins and histamine for the occurrence of shock during acute hemorrhagic pancreatitis is pointed out.

Topics: Acute Disease; Chymotrypsin; Enzyme Activation; Enzyme Precursors; Fat Necrosis; Histamine Release; Humans; Kallikreins; Kinins; Pancreatic Elastase; Pancreatitis; Peptide Hydrolases; Phospholipases; Shock, Hemorrhagic; Triglycerides; Trypsin; Trypsinogen

Acute pancreatitis was produced in five dogs by injecting bile into the pancreatic duct. The capillary permeability effects of the exudate formed within the peritoneal cavity were studied by injecting the exudate intradermally into puppies. The amount of radioactively labeled albumin escaping from the circulation and appearing at the intradermal injection site was used as a measure of capillary permeability. It was observed that the peritoneal exudate, especially that produced in the early stage of bile induced pancreatitis, contains one or more substances which result in an increased capillary permeability when injected intradermally into puppies.

Topics: Amylases; Animals; Ascitic Fluid; Capillary Permeability; Dogs; Female; Lipase; Male; Pancreatic Elastase; Pancreatitis; Trypsin; Trypsinogen

Topics: Alcoholism; Humans; Pancreatic Juice; Pancreatitis; Trypsin; Trypsin Inhibitors; Trypsinogen

The trypsinogen and chymotrypsinogen contents of the pancreas were examined during acute experimental pacreatitiis of the rat. The proenzymes were activated with enterokinase and the amounts of active proteases were estimated with BAPNA (N-alfa-benzoyl-DL-arginin-4-nitroanilid hydrochlorid, Fluka AG) and SUPHEPA (succinyl-L-phenylalanine-p-nitroanilide, Schwarz/Mann, Division of Becton) as the substrates. The activation of chymotrypsinogen was more rapid than the activation of trypsinogen; maximal activation occurred in 3 hours. Under similar circumstances the activation of trypsinogen required 17 hours. Both trypsinogen and chymotrypsinogen content decreased significantly during the inflammation. In 8 hours the decline of trypsinogen content was 28.4 percent and that of chymotrypsinogen content 44.9 percent from the proenzyme content of the normal resting rat pancreas. This indicates that proenzymes and/or active proteases are liberated during the course of pancreatitis. No correlation was found between the trypsinogen and the chymotrypsinogen content of the normal pancreas, but during pancreatitis the proenzyme contents correlated clearly. The correlation during inflammation possibly reflects the amount of the viable pancreatic tissue and the rate of synthesis.

Topics: Acute Disease; Animals; Buffers; Chymotrypsinogen; Enteropeptidase; Enzyme Activation; Pancreatic Juice; Pancreatitis; Peptide Hydrolases; Rats; Stimulation, Chemical; Time Factors; Trypsinogen

The changes in elastase levels were studied in the plasma before and after bile-induced pancreatitis. Although the anylase and lipase levels increased markedly, there was no significant increase in the plasma elastase levels, as measured by enzymatic assay. On the other hand, the elastase levels of the blood in the femoral and pancreatic veins as measured by radioimmunoassay, increased to about ten times above the control levels. Similarly, in pancreatitis, large amounts of elastase, determined by radioimmunoassay, was found in the ascitic fluid. At that time, no elastase activity could be determined in the ascitic fluid. In this study, it is suggested that circulating inhibitors interfere with the determination of elastase enzymatic activity but do not interfere with the radioimmunoassay of elastase.

Topics: Acute Disease; Amylases; Animals; Ascitic Fluid; Bile; Dogs; Humans; Lipase; Pancreatic Elastase; Pancreatitis; Radioimmunoassay; Trypsin; Trypsinogen

Topics: Acute Disease; Adult; Aged; Catheterization; Chymotrypsin; Chymotrypsinogen; Enzyme Activation; Enzyme Precursors; Female; Humans; Hydrogen-Ion Concentration; Male; Middle Aged; Pancreatic Ducts; Pancreatic Elastase; Pancreatic Fistula; Pancreatic Juice; Pancreatitis; Proteins; Temperature; Thrombin; Time Factors; Trypsin; Trypsinogen

Topics: Angiotensin II; Arginine; Blood Coagulation Disorders; Blood Coagulation Tests; Blood Proteins; Caseins; Chromatography, Gel; Chymotrypsinogen; Enzyme Activation; Esters; Fibrin; Fibrinogen; Hemoglobins; Humans; Hydrolysis; Macroglobulins; Pancreatitis; Protein Binding; Protein Denaturation; Thrombin; Trypsin; Trypsin Inhibitors; Trypsinogen; Vasopressins

Topics: Amylases; Animals; Ascitic Fluid; Blood Pressure; Bradykinin; Dogs; Enzyme Induction; Enzymes; Hypotension; Kinins; Lipase; Pancreatic Juice; Pancreatitis; Regional Blood Flow; Trypsin; Trypsinogen

Topics: Amylases; Animals; Ascitic Fluid; Chymotrypsin; Dogs; Female; Immunoassay; Macroglobulins; Male; Pancreatitis; Protein Binding; Shock; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Amylases; Autopsy; Bone Marrow Diseases; Child, Preschool; Clinical Enzyme Tests; Copper; Endopeptidases; Female; Humans; Infant; Kwashiorkor; Lipase; Male; Metabolism, Inborn Errors; Pancreas; Pancreatic Diseases; Pancreatitis; Trypsinogen; Zinc

Topics: Amylases; Animals; Ascitic Fluid; Blood Glucose; Blood Pressure; Dogs; Female; Femoral Vein; Hyperglycemia; Insulin; Insulin Antagonists; Insulin Secretion; Lipase; Lymph; Male; Pancreatic Elastase; Pancreatitis; Peptide Hydrolases; Portal Vein; Radioimmunoassay; Time Factors; Trypsin; Trypsinogen

Topics: Animals; Bile Acids and Salts; Blood Coagulation; Cattle; Esterases; Fibrinogen; Fibrinolysin; Fibrinolysis; Hemostasis; Humans; Pancreatic Juice; Pancreatitis; Plasminogen; Platelet Adhesiveness; Protein Binding; Prothrombin; Surface Properties; Thrombelastography; Thrombin; Trypsin; Trypsinogen

Topics: Acute Disease; Adenoma; Amylases; Biliary Tract Surgical Procedures; Colon; Humans; Pancreatitis; Parathyroid Neoplasms; Rectum; Stomach; Thyroidectomy; Trypsin; Trypsinogen

Topics: Animals; Dogs; Ethanol; Pancreas; Pancreatic Juice; Pancreatitis; Perfusion; Secretin; Stimulation, Chemical; Time Factors; Trypsin Inhibitors; Trypsinogen

Topics: Animals; In Vitro Techniques; Male; Methods; Pancreas; Pancreatitis; Rats; Trypsin; Trypsinogen

Topics: Alcoholism; Amylases; Animals; Dietary Carbohydrates; Dietary Fats; Dietary Proteins; Ethanol; Humans; Lipase; Male; Pancreas; Pancreatic Juice; Pancreatitis; Rats; Trypsinogen

Topics: Acute Disease; Amylases; Animals; Dogs; Humans; Injections; Lipase; Pancreatic Juice; Pancreatitis; Phospholipases; Trypsin; Trypsinogen

Topics: Acute Disease; Humans; Pancreatic Juice; Pancreatitis; Trypsin; Trypsinogen

Topics: Acetates; Albumins; Amylases; Carboxypeptidases; Cellulose; Chymotrypsin; Duodenum; Electrophoresis; Enzyme Precursors; Humans; Intestinal Secretions; Lipase; Pancreatic Juice; Pancreatitis; Ribonucleases; Trypsin Inhibitors; Trypsinogen

Topics: Amylases; Animals; Cholecystokinin; Diet; Dogs; Electrophoresis, Disc; Lipase; Pancreas; Pancreatic Juice; Pancreatitis; Proteins; Secretin; Trypsinogen

Topics: Animals; Diet; Dogs; Pancreas; Pancreatitis; Time Factors; Trypsinogen

Topics: Acute Disease; Agar; Amino Acid Sequence; Animals; Aprotinin; Blood Protein Electrophoresis; Cats; Dogs; Female; Gels; Male; Models, Biological; Pancreatic Juice; Pancreatitis; Trypsinogen

Topics: Acute Disease; Amylases; Animals; Blood; Carboxypeptidases; Chymotrypsin; Culture Techniques; Dogs; Enzymes; Lipase; Pancreatic Juice; Pancreatitis; Phospholipases; Trypsin; Trypsinogen

Topics: Animals; Bile; Bile Acids and Salts; Dogs; Gallbladder; Goats; Metabolism; Pancreatic Juice; Pancreatitis; Physiology; Salts; Sheep; Trypsin; Trypsinogen

Topics: Animals; Cholecystokinin; Dogs; Enzyme Precursors; Gastrointestinal Hormones; Pancreas; Pancreatitis; Secretin; Trypsinogen

Topics: Aprotinin; Bile Acids and Salts; Enzyme Inhibitors; Kallikreins; Pancreatitis; Pharmacology; Rats; Research; Salts; Sodium Chloride; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Bile; Pancreas; Pancreatitis; Peptide Hydrolases; Trypsinogen

Topics: Diatrizoate; Disease; Humans; Hydrolases; Pancreas; Pancreatic Cyst; Pancreatic Ducts; Pancreatic Juice; Pancreatitis; Pharmacology; Radiography; Trypsin; Trypsinogen

Topics: Amylases; Aprotinin; Blood Chemical Analysis; Clinical Enzyme Tests; Kallikreins; Pancreatitis; Peptide Hydrolases; Rats; Research; Trypsin; Trypsinogen

Topics: Humans; Pancreatitis; Trypsin; Trypsinogen

Topics: Disease; Humans; Pancreatic Juice; Pancreatitis; Parathyroid Diseases; Parathyroid Glands; Trypsin; Trypsin Inhibitors; Trypsinogen

Topics: Enzyme Precursors; Pancreas; Pancreatitis; Trypsin; Trypsinogen