Cathepsin B-Mediated Activation of Trypsinogen in Endocytosing Macrophages Increases Severity of Pancreatitis in Mice.

BACKGROUND & AIMS: Acute pancreatitis is characterized by premature intracellular activation of digestive proteases within pancreatic acini and a consecutive systemic inflammatory response. We investigated how these processes interact during severe pancreatitis in mice. METHODS: Pancreatitis was induced in C57Bl/6 wild-type (control), cathepsin B (CTSB)-knockout, and cathepsin L-knockout mice by partial pancreatic duct ligation with supramaximal caerulein injection, or by repetitive supramaximal caerulein injections alone. Immune cells that infiltrated the pancreas were characterized by immunofluorescence detection of Ly6g, CD206, and CD68. Macrophages were isolated from bone marrow and incubated with bovine trypsinogen or isolated acinar cells; the macrophages were then transferred into pancreatitis control or cathepsin-knockout mice. Activities of proteases and nuclear factor (NF)-κB were determined using fluorogenic substrates and trypsin activity was blocked by nafamostat. Cytokine levels were measured using a cytometric bead array. We performed immunohistochemical analyses to detect trypsinogen, CD206, and CD68 in human chronic pancreatitis (n = 13) and acute necrotizing pancreatitis (n = 15) specimens. RESULTS: Macrophages were the predominant immune cell population that migrated into the pancreas during induction of pancreatitis in control mice. CD68-positive macrophages were found to phagocytose acinar cell components, including zymogen-containing vesicles, in pancreata from mice with pancreatitis, as well as human necrotic pancreatic tissues. Trypsinogen became activated in macrophages cultured with purified trypsinogen or co-cultured with pancreatic acini and in pancreata of mice with pancreatitis; trypsinogen activation required macrophage endocytosis and expression and activity of CTSB, and was sensitive to pH. Activation of trypsinogen in macrophages resulted in translocation of NF-kB and production of inflammatory cytokines; mice without trypsinogen activation (CTSB-knockout mice) in macrophages developed less severe pancreatitis compared with control mice. Transfer of macrophage from control mice to CTSB-knockout mice increased the severity of pancreatitis. Inhibition of trypsin activity in macrophages prevented translocation of NF-κB and production of inflammatory cytokines. CONCLUSIONS: Studying pancreatitis in mice, we found activation of digestive proteases to occur not only in acinar cells but also in macrophages that infiltrate pancreatic tissue. Activation of the proteases in macrophage occurs during endocytosis of zymogen-containing vesicles, and depends on pH and CTSB. This process involves macrophage activation via NF-κB-translocation, and contributes to systemic inflammation and severity of pancreatitis.

Inhibition of 5-lipoxygenase represses neutrophils activation and activates apoptosis in pancreatic tissues during acute necrotizing pancreatitis.

Pancreatic glandular necrosis is rapid inflammation of the pancreas and contributes to severe acute pancreatitis in humans. The pathogenesis of pancreatic tissue inflammation during acute pancreatitis is still largely unknown. Recent studies suggest that 5-lipoxygenase (5-LOX) is an essential mediator in modulating cell death pathways in human diseases. In this study, we aimed to evaluate the effects of a 5-LOX inhibitor, zileuton, on tissue apoptosis and neutrophils activation in pancreatic tissues during acute necrotizing pancreatitis (ANP) in a rat model. In this present study, both mRNA and protein levels of 5-LOX are upregulated during ANP and zileuton treatment is shown to repress ANP-induced upregulation of 5-LOX levels. In addition, zileuton treatment is found to repress blood biomarkers of neutrophils activation such as soluble intercellular adhesive molecular 1 (ICAM-1), soluble E-selectin (E-selectin), soluble P-selectin (P-selectin), leukotriene B4 (LTB4), and myeloperoxidase (MPO). Also, zileuton treatment attenuates pancreatic tissue pathology, upregulates caspase-3, downregulates B-cell lymphoma 2 (Bcl-2), and activates tissue apoptosis evaluated by TUNEL staining. Our results show that 5-LOX plays an important role in activating apoptosis and repressing neutrophils activation during ANP. The current study suggests that 5-LOX can be used as a potential target for the treatment of ANP.

Underexpression of Receptor for Activated C Kinase 1 (RACK1) in Leukocytes from Patients with Severe Acute Pancreatitis.

Receptor for activated C kinase 1 (RACK1) plays an important role in regulating the immune response and cytokine expression. However, little is known about its role in acute pancreatitis (AP). We therefore investigated the role of RACK1 in AP and explored its relationship with interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), both of which are related to AP severity. Two rat models of chemically induced AP with different severities were used: acute edematous pancreatitis (AEP) and acute necrotizing pancreatitis (ANP). The expression levels of IL-6 and TNF-α mRNAs and proteins were significantly increased in leukocytes from AEP and ANP rats, compared with the levels in the control animals, while the expression levels of RACK1 mRNA and protein were significantly decreased in leukocytes from these AP rats. Moreover, the RACK1 levels in leukocytes were significantly lower in ANP rats than those in AEP rats. Consequently, AP patients and healthy volunteers (HVs) were enrolled in this study. Compared with the HVs (n = 5), the expression levels of IL-6 and TNF-α mRNAs and proteins were significantly higher in leukocytes from 15 AP patients, including patients with mild AP (n = 5). By contrast, the expression levels of RACK1 mRNA and protein in leukocytes were significantly lower among patients with severe AP (n = 5) and with moderately severe AP (n = 5), compared with the HVs. The expression levels of RACK1 mRNA were negatively correlated with the IL-6 and TNF-α mRNA levels. Thus, RACK1 may alleviate the severity of AP.

Serum C-reactive protein, procalcitonin, and lactate dehydrogenase for the diagnosis of pancreatic necrosis.

BACKGROUND: The treatment of people with pancreatic necrosis differs from that of people with oedematous pancreatitis. It is important to know the diagnostic accuracy of serum C-reactive protein (CRP), serum procalcitonin, and serum lactate dehydrogenase (LDH) as a triage test for the detection of pancreatic necrosis in people with acute pancreatitis, so that an informed decision can be made as to whether the person with pancreatic necrosis needs further investigations such as computed tomography (CT) scan or magnetic resonance imaging (MRI) scan and treatment for pancreatic necrosis started. There is currently no standard clinical practice, although CRP, particularly an increasing trend of CRP, is often used as a triage test to determine whether the person requires further imaging. There is also currently no systematic review of the diagnostic test accuracy of CRP, procalcitonin, and LDH for the diagnosis of pancreatic necrosis in people with acute pancreatitis. OBJECTIVES: To compare the diagnostic accuracy of CRP, procalcitonin, or LDH (index test), either alone or in combination, in the diagnosis of necrotising pancreatitis in people with acute pancreatitis and without organ failure. SEARCH METHODS: 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. SELECTION CRITERIA: We included all studies that evaluated the diagnostic test accuracy of CRP, procalcitonin, and LDH for the diagnosis of pancreatic necrosis in people with acute pancreatitis using the following reference standards, either alone or in combination: radiological features of pancreatic necrosis (contrast-enhanced CT or MRI), surgeon's judgement of pancreatic necrosis during surgery, or histological confirmation of pancreatic necrosis. Had we found case-control studies, we planned to exclude them because they are prone to bias; however, we did not locate any. Two review authors independently identified the relevant studies from the retrieved references. DATA COLLECTION AND ANALYSIS: Two review authors independently extracted data, including methodological quality assessment, from the included studies. As the included studies reported CRP, procalcitonin, and LDH on different days of admission and measured at different cut-off levels, it was not possible to perform a meta-analysis using the bivariate model as planned. We have reported the sensitivity, specificity, post-test probability of a positive and negative index test along with 95% confidence interval (CI) on each of the different days of admission and measured at different cut-off levels. MAIN RESULTS: A total of three studies including 242 participants met the inclusion criteria for this review. One study reported the diagnostic performance of CRP for two threshold levels (> 200 mg/L and > 279 mg/L) without stating the day on which the CRP was measured. One study reported the diagnostic performance of procalcitonin on day 1 (1 day after admission) using a threshold level of 0.5 ng/mL. One study reported the diagnostic performance of CRP on day 3 (3 days after admission) using a threshold level of 140 mg/L and LDH on day 5 (5 days after admission) using a threshold level of 290 U/L. The sensitivities and specificities varied: the point estimate of the sensitivities ranged from 0.72 to 0.88, while the point estimate of the specificities ranged from 0.75 to 1.00 for the different index tests on different days of hospital admission. However, the confidence intervals were wide: confidence intervals of sensitivities ranged from 0.51 to 0.97, while those of specificities ranged from 0.18 to 1.00 for the different tests on different days of hospital admission. Overall, none of the tests assessed in this review were sufficiently accurate to suggest that they could be useful in clinical practice. AUTHORS' CONCLUSIONS: The paucity of data and methodological deficiencies in the studies meant that it was not possible to arrive at any conclusions regarding the diagnostic test accuracy of the index test because of the uncertainty of the results. Further well-designed diagnostic test accuracy studies with prespecified index test thresholds of CRP, procalcitonin, LDH; appropriate follow-up (for at least two weeks to ensure that the person does not have pancreatic necrosis, as early scans may not indicate pancreatic necrosis); and clearly defined reference standards (of surgical or radiological confirmation of pancreatic necrosis) are important to reliably determine the diagnostic accuracy of CRP, procalcitonin, and LDH.

Predicting severe acute pancreatitis in children based on serum lipase and calcium: A multicentre retrospective cohort study.

OBJECTIVE: This study aims to identify predictors of severe paediatric AP based on laboratory trends and peak/trough values on day 2 (D2) after presentation. The performance of identified predictors was first assessed and then combined with the previously validated sensitive predictor serum lipase ≥7 times the upper limit of normal (× ULN) on day 1 (D1). METHODS: A retrospective review of children with AP (January 2000-July 2011) was performed at three tertiary referral hospitals (two in Australia, one in the Netherlands). Trends of candidate predictors were analysed using the percentage change from D1 to D2 or peak/trough values within 48 h after presentation. RESULTS: 175 AP episodes (including 50 severe episodes [29%]) were identified. Serum lipase ≥50% decrease on D2 (sensitivity 73%, specificity 54%) and calcium trough ≤2.15 mmol/L within 48 h (sensitivity 59%, specificity 81%) were identified as statistically significant predictors for severe AP. By combining the newly identified predictors with the previously validated predictor serum lipase ≥7× ULN on D1 (sensitivity 82%, specificity 53%), specificity improved to predict severe AP on D2 with the addition of: (i) serum lipase ≥50% decrease (sensitivity 67%, specificity 79%), or (ii) trough calcium ≤2.15 mmol/L (sensitivity 46%, specificity 89%). CONCLUSIONS: Serum lipase and calcium, may be helpful in predicting severity of paediatric AP. There may be a clinical role on D1 for using serum lipase ≥7× ULN (high sensitivity), and on D2 for combining D1 serum lipase ≥7× ULN with calcium trough ≤2.15 mmol/L within 48 h (high specificity) to help predict severe paediatric AP.

Effect of Chaiqinchengqi decoction on inositol requiring enzyme 1α in alveolar macrophages of dogs with acute necrotising pancreatitis induced by sodium taurocholate.

OBJECTIVE: To investigate the effect of Chaiqinchengqi decoction (CQCQD) on inositol requiring enzyme lα (IRElα) in alveolar macrophages (AMs) of the dog model of acute necrotising pancreatitis (ANP) induced by sodium taurocholate. METHODS: Fifteen beagle dogs were randomised into a control group, ANP group and CQCQD group (n = 5 per group). ANP was induced by a retrograde duct injection of 50 mg/kg of 5% sodium taurocholate. The dogs in the control group received injections of the same volume of saline as the sodium taurocholate. After the models were induced, the dogs in the CQCQD group were administered 10 mL/kg CQCQD every 2 h for 6 h. Two hours after the last administration of either CQCQD or saline, they were sacrificed by anaesthesia. AMs were collected to determine the IRElα and interleukin-1ß (IL-1ß) mRNA and protein expression, and pancreatic tissues were collected for histopathology analysis. RESULTS: Compared with the ANP group, the mRNA and protein expression of IREl a and the protein expression of IL-1ß of AMs in the CQCQD group were significantly down-regulated, and the pancreatic histopathology score of the CQCQD group also was lower. There was no significant difference in the mRNA expression of IL-1ß of AMs between the two groups. CONCLUSION: The CQCQD-induced down-regulation of the IL-1ß protein expression may involve the down-regulation of the mRNA and protein expression of IRElα in AMs.

Tumour necrosis factor α secretion induces protease activation and acinar cell necrosis in acute experimental pancreatitis in mice.

BACKGROUND: Acute pancreatitis has long been considered a disorder of pancreatic self-digestion, in which intracellular activation of digestive proteases induces tissue injury. Chemokines, released from damaged pancreatic cells then attract inflammatory cells, whose systemic action ultimately determines the disease severity. In the present work the opposite mechanism is investigated; that is, whether and how inflammatory cells can activate intracellular proteases. DESIGN: Using mice either deficient for the CD18-α subunit of the membrane attack complex-1 (MAC-1) complex or tumour necrosis factor (TNF)α, as well as after depletion of leucocyte subpopulations, pancreatitis was induced by 7-hourly caerulein injections (50 µg/kg, intraperitoneally). Pancreatic acini were coincubated in vitro from wild-type and cathepsin-B-deficient animals with phorbol-12-myristate-13-acetate (PMA)-activated neutrophils and macrophages, caerulein or TNFα, and activities of trypsin, cathepsin-B and caspase-3 were measured, as well as necrosis using fluorogenic substrates. TNFα was inhibited with monospecific antibodies. RESULTS: Deletion of CD18 prevented transmigration of leucocytes into the pancreas during pancreatitis, greatly reduced disease severity and abolished digestive protease activation. Depletion of neutrophils and macrophages equally reduced premature trypsinogen activation and disease severity. In vitro activated neutrophils and macrophages directly induced premature protease activation and cell death in pancreatic acini and stimulation of acini with TNFα induced caspase-3 activation and necrosis via a cathepsin-B and calcium-dependent mechanism. Neutralising antibodies against TNFα and genetic deletion of TNFα prevented leucocyte-induced trypsin activity and necrosis in isolated acini. CONCLUSIONS: The soluble inflammatory cell mediator TNFα directly induces premature protease activation and necrosis in pancreatic acinar cells. This activation depends on calcium and cathepsin-B activity. The findings from the present work further suggest that targeting TNFα, for which pharmaceutical agents are readily available, could be an effective treatment strategy that directly addresses the cellular causes of pancreatitis.

The role of apigenin in an experimental model of acute pancreatitis.

AIM: The aim of the present study is to evaluate pathologic changes in the pancreatic parenchyma in an experimental model of acute pancreatitis (AP) following bilio-pancreatic duct ligation. An effort was made to clarify the role of apigenin, a substance that is well-known for its antioxidant and anti-inflammatory role and its likely beneficial activity to the pancreatic parenchyma following AP in rats. MATERIAL AND METHOD: One hundred twenty-six male Wistar rats 3-4 mo old and weighing 220-350 g were used. At time 0, the following groups were randomly assigned: group sham: rats were subjected to virtual surgery; group control: rats were subjected to surgery for induction of AP, by ligation of the bilio-pancreatic duct; group apigenin: rats were subjected to surgery for induction of AP and enteral feeding with apigenin. Pathologic changes of the pancreatic parenchymal and myeloperoxidase activity were measured at predetermined time intervals 6, 12, 24, 48, and 72 h. RESULT: From the pathologic reports, by comparing the control group with the apigenin group, an improvement of pancreatic tissue architecture following apigenin administration was observed. Inflammatory infiltration, edema, ductal dilation, and necrosis were reduced following apigenin administration over time (P = 0.049, P = 0.228, P = 0.387, P = 0.046). Treatment with apigenin significantly reduced the bilio-pancreatic duct ligation and evoked an increase in pancreatic myeloperoxidase activity (P = 0.030). CONCLUSION: Oral apigenin administration in rats, following experimentally induced pancreatitis, seems to protect the pancreatic tissue. Thus, apigenin administration to humans could potentially ameliorate the damages to the pancreas.

The role of neutral endopeptidase in caerulein-induced acute pancreatitis.

Substance P (SP) is well known to promote inflammation in acute pancreatitis (AP) by interacting with neurokinin-1 receptor. However, mechanisms that terminate SP-mediated responses are unclear. Neutral endopeptidase (NEP) is a cell-surface enzyme that degrades SP in the extracellular fluid. In this study, we examined the expression and the role of NEP in caerulein-induced AP. Male BALB/c mice (20-25 g) subjected to 3-10 hourly injections of caerulein (50 µg/kg) exhibited reduced NEP activity and protein expression in the pancreas and lungs. Additionally, caerulein (10(-7) M) also downregulated NEP activity and mRNA expression in isolated pancreatic acinar cells. The role of NEP in AP was examined in two opposite ways: inhibition of NEP (phosphoramidon [5 mg/kg] or thiorphan [10 mg/kg]) followed by 6 hourly caerulein injections) or supplementation with exogenous NEP (10 hourly caerulein injections, treatment of recombinant mouse NEP [1 mg/kg] during second caerulein injection). Inhibition of NEP raised SP levels and exacerbated inflammatory conditions in mice. Meanwhile, the severity of AP, determined by histological examination, tissue water content, myeloperoxidase activity, and plasma amylase activity, was markedly better in mice that received exogenous NEP treatment. Our results suggest that NEP is anti-inflammatory in caerulein-induced AP. Acute inhibition of NEP contributes to increased SP levels in caerulein-induced AP, which leads to augmented inflammatory responses in the pancreas and associated lung injury.

Activity of neutrophil elastase reflects the progression of acute pancreatitis.

OBJECTIVE: Neutrophil elastase (NE) concentration is associated with progression of acute pancreatitis (AP), but measuring total NE concentration includes biologically inactive NE. This study aims to investigate the relationship between NE activity and the aetiology and severity of AP and associated organ failure. METHODS: Seventy-five patients admitted to our surgery department with a first episode of AP during 2004-2005 were age- and sex-matched to 20 healthy volunteers (controls). NE activity was assessed using venous blood samples obtained on patient admission and after 1, 2 and 14 days. One sample was also taken from each control. ANOVA was used for statistical comparison between groups. RESULTS: Baseline NE activity (geometric mean; 95% confidence intervals) differed between patients (58.6 nM of substrate 7-amino-4-methylcoumarin [AMC]/hour; 48.52-70.72) and controls (31.5 nM AMC/hour; 25.5-39.0) (p = 0.0003), and did not correlate with time between symptom onset and admission. Patients with alcohol-induced AP demonstrated higher mean activity (59.1 nM AMC/h; 44.7-78.2) than those with gallstone-induced AP (41.7 nM AMC/h; 33.9-51.4) (p = 0.0496). NE activity was higher overall in patients with predicted severe AP (60.9 nM AMC/h; 48.0-77.2) than in those with predicted mild AP (42.1 nM AMC/h; 34.9-50.8) (p = 0.027). Patients with respiratory failure had higher NE activity (82.5 nM AMC/h; 57.5-118.4) than those without (43.9 nM AMC/h; 37.6-51.3) (p = 0.0024). CONCLUSIONS: NE activity was associated with predicted severity of AP and AP-associated respiratory failure. Specific NE inhibitors may have therapeutic potential in acute pancreatitis.

Prospective, randomized, double-blind, placebo-controlled trial of ulinastatin for prevention of hyperenzymemia after double balloon endoscopy via the antegrade approach.

BACKGROUND: Double balloon endoscopy (DBE) allows the entire small intestine to be viewed using a combination of antegrade and retrograde approaches. Acute pancreatitis is a serious complication of antegrade DBE with no effective prophylactic treatment currently available. Ulinastatin has been shown to be effective for the prevention of pancreatitis following endoscopic retrograde cholangiopancreatography. We therefore assessed the efficacy of ulinastatin for hyperenzymemia after antegrade DBE. PATIENTS AND METHODS: Forty-four patients were enrolled in this prospective, randomized, double-blind, placebo-controlled trial. Patients in the ulinastatin group received 150 000 U ulinastatin by i.v. drip infusion for 2 h from the start of the procedure. Serum concentrations of pancreatic amylase and lipase were measured before and 3 and 18 h after antegrade DBE. RESULTS: The study was terminated after interim analysis. Of the 44 patients, 23 were randomized to ulinastatin and 21 to placebo.The groups were similar with regard to sex ratio, age, type of endoscope, insertion time, total procedure time, number of endoscope pull-back procedures, and baseline pancreaticamylase and lipase concentrations. Post-DBE hyperenzymemia was observed in 35.0% and 47.8% of patients in the placebo and ulinastatin groups, respectively. The higher frequency of hyperenzymemia in the ulinastatin group was unexpected, but the difference was not statistically significant. One patient in the placebo group (5.0%) and none in the ulinastatin group experienced acute pancreatitis, but the difference was not statistically significant. CONCLUSION: The results of this trial suggest that ulinastatin does not prevent hyperenzymemia following antegrade DBE.

[The efficacy of the acute pancreatitis' surgical treatment].

The comparative analysis of blood levels of leukocytes, lymphocytes, the leukocytic intoxication index, amylase, lipase, lactatdehydrogenase and creatinphosphokinase, measured in operated patients with the acute pancreatitis, demonstrated the general positive dynamics of the patients condition. The higher blood levels of the substances in died patients demonstrate the important prognostic value of them. The higher levels of amylase, lipase, lactatdehydrogenase and creatinphosphokinase by the end of the clinical treatment together with the normalization of the rest laboratory data may witness the higher risk of the chronisation of the pancreatitis.

[Some of criteria in the evaluation of severity and prognosis with different forms of acute pancreatitis].

UNLABELLED: The aim of the research is to improve diagnosis and assessment of severity and prognosis with different forms of acute pancreatitis. In 79 patients were studied levels of leukocytes, lymphocytes, leucocyte intoxication index (LII), the content of lactate dehydrogenase (LDG), creatinephosphokinase (CPK), amylase, aspartate aminotransferase (AST, alanine aminotransferase (ALT). RESULTS: The level of leukocytes reflects the severity of the disease, but had no prognostic value. The level of lymphocytes, LII. LDG and lipase reflect the severity, of the disease and have prognostic value. The level of amylase, AST, ALT, CPK not always reflect the severity of the disease, but had prognostic value. CONCLUSION: The most readily available to assess the severity and prognosis in acute pancreatitis are the level of blood lymphocytes and LII. Indicators LDG. CPK, amylase, lipase, AST and ALT also reflect the course and prognosis of the disease.

Intracellular activation of trypsinogen in transgenic mice induces acute but not chronic pancreatitis.

BACKGROUND AND AIMS: Premature intra-acinar activation of trypsinogen is widely considered key for both the initiation of acute pancreatitis and the development of chronic pancreatitis. However, the biological consequences of intracellular trypsinogen activation have not been directly examined. To do so, a new mouse model was developed. METHODS: Mice were engineered to conditionally express an endogenously activated trypsinogen within pancreatic acinar cells (PACE-tryp(on)). Hallmarks of pancreatitis were determined and findings were correlated to the level (zygosity) and extent (temporal and spatial) of conditional PACE-tryp(on) expression. Furthermore, the impact of acinar cell death in PACE-tryp(on) mice was assessed and compared with a model of selective diphtheria toxin (DT)-mediated induction of acinar apoptosis. RESULTS: Initiation of acute pancreatitis was observed with high (homozygous), but not low (heterozygous) levels of PACE-tryp(on) expression. Subtotal (maximal-rapid induction) but not limited (gradual-repetitive induction) conditional PACE-tryp(on) expression was associated with systemic complications and mortality. Rapid caspase-3 activation and apoptosis with delayed necrosis was observed, and loss of acinar cells led to replacement with fatty tissue. Chronic inflammation or fibrosis did not develop. Selective depletion of pancreatic acinar cells by apoptosis using DT evoked similar consequences. CONCLUSIONS: Intra-acinar activation of trypsinogen is sufficient to initiate acute pancreatitis. However, the primary response to intracellular trypsin activity is rapid induction of acinar cell death via apoptosis which facilitates resolution of the acute inflammation rather than causing chronic pancreatitis. This novel model provides a powerful tool to improve our understanding of basic mechanisms occurring during pancreatitis.

Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) activity in the sera of patients with acute and chronic pancreatitis.

OBJECTIVE: Acute and chronic pancreatitis is a major complication of alcohol abuse. The pancreas can metabolize ethanol via oxidative pathway involving the enzymes - alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) as well as the nonoxidative pathway. Human pancreas tissue contains various ADH isoenzymes and possesses also ALDH activity. In this paper we have measured the activity of alcohol dehydrogenase isoenzymes, and aldehyde dehydrogenase in the sera of patients with acute and chronic pancreatitis. METHODS: Serum samples were taken for routine biochemical investigation from 46 patients suffering from acute pancreatitis and 32 patients with chronic pancreatitis. Total ADH activity was measured by photometric method with p-nitrosodimethylaniline (NDMA) as a substrate and ALDH activity by the fluorometric method with 6-methoxy-2-naphtaldehyde as a substrate. For the measurement of the activity of class I isoenzymes we employed the fluorometric methods, with class-specific fluorogenic substrates. The activity of class III alcohol dehydrogenase was measured by the photometric method with n-octanol and class IV with m-nitrobenzaldehyde as a substrate. RESULTS: A statistically significant increase of class III alcohol dehydrogenase isoenzymes was found in the sera of patients with acute and chronic pancreatitis. The median activity of this class isoenzyme in the patients group increased about 35% in the comparison to the control level. The total alcohol dehydrogenase activity was also significantly higher (23.5%) among patients with pancreatitis than healthy ones. The activities of other tested ADH isoenzymes and total ALDH were unchanged. The activity of the class I ADH isoenzyme was significantly higher in the sera of heavy drinkers with pancreatitis. CONCLUSION: We can state that the increase of the activity of class III alcohol dehydrogenase isoenzyme in the sera of pancreatitis patients seems to be caused by the release of this isoenzyme from damaged pancreatic cells.

[Criteria determining indications to operative treatment of purulent complications of acute pancreatitis in patients who had endured endovideosurgical interventions at the enzymatic phase].

An analysis of 72 criteria was made which include 5 main (hectic fever, tachycardia, leukocytosis with a neutrophilic shift to the left, decreased index of the relation of total protein to the amount of values of urea and creatinin less than 0.94 c.u., male sex), 3 additional (anemization, hypoproteinemia, decreased index of the relation of temperature to pulse rate less than 0.5 c.u.) and 1 determinant (the presence of liquid in the abdominal cavity and/or in the omental bursa and/or in the retroperitoneal space. In the presence of three of the main and one of additional signs the indications to operation are determined with probability 90.2%, and in the presence of three main and one additional and determinant--100%.

Inflammatory rather than infectious insults play a role in exocrine tissue damage in a mouse model for coxsackievirus B4-induced pancreatitis.

Infection with coxsackievirus B4 (CVB4) may result in an acute severe necrotizing pancreatitis that mostly remains restricted to the acini of the exocrine parenchyma. The mechanisms responsible for this tissue damage, however, remain poorly understood. We here report that COAM, a polyanionic carboxylic acid, provides marked protection against CVB4-induced pancreatitis in a mouse model. Despite the fact that COAM largely reduced disease severity, as detected by serum amylase and lipase levels as well as histologically, titres of replicating CVB4 in the pancreas were virtually unaffected. COAM treatment diminished the infection-associated MMP-9 levels and also resulted in a decreased influx of neutrophils into the infected pancreas. Moreover, we demonstrate that titres of replicating virus in the pancreas did not directly correlate with the severity of disease. In conclusion, our data suggest that immunopathological effects, rather than direct virus-induced destruction, are responsible for the damage to acinar tissue in CVB4-induced pancreatitis.

Serum fetuin A concentrations in patients with acute pancreatitis.

BACKGROUND: Acute pancreatitis (AP) is a mild and self-limiting disease in most patients, but necrotizing pancreatitis develops in up to 20 - 30% of the cases. Early recognition of severe AP has been considered as a key determinant of successful therapy. The aim of this study was to evaluate the clinical value of fetuin A as the new predictor of complications and fatal outcome during acute pancreatitis (AP). METHODS: The study included 40 patients with AP of diverse severity (28 mild, 12 severe), assessed during the early phase of AP (1st - 7th day of hospitalization). Fetuin A level was measured by ELISA kit (BioVendor). RESULTS: Median serum fetuin A concentrations had a tendency to decrease during examination from 0,371 g/L at admission to 0,288 g/L on the 7th day of hospitalization. In each of 7 days of observation, correlation between the increase in fetuin A and the absolute number of RBC was found (R = 0,34: 1st day R = 0,35: 3rd day, p < 0,05; R = 0,57: 5th day, p < 0,001; R = 0,65: 7th day, p < 0,01). Additionally, we observed the reverse relationship between the decrease in fetuin A and the increase in some inflammatory markers (IL-6: R = -0,61, p < 0,0001; SAA: R = -0,58, p < 0,001; HGF: R = -0,60, p < 0,01; PCT: R = -0,475, p < 0,01). The strongest positive correlation was noticed on the 5th day of hospitalization between decreased levels of fetuin A and albumin (R = 0,83; p < 0,0001). CONCLUSIONS: Fetuin A level monitoring is potentially a new marker for non-invasive and accurate prediction status for the hospitalization of patients with AP similar to other negative acute phase proteins like albumin.

Acute pancreatitis due to rotavirus gastroenteritis in a child.

OBJECTIVES: Serum amylase can be elevated in various pathological conditions, but its primary application is as a biochemical test for the diagnosis of acute pancreatitis. Acute gastroenteritis has not been widely recognized as a cause for hyperamylasemia and pancreatitis. METHODS: We describe a case of acute pancreatitis after rotavirus gastroenteritis occurring in a previously healthy child. RESULTS: Rotavirus infection was demonstrated by detection of virus particles in stool culture and pancreatitis by raised serum amylase and lipase levels. No abnormalities were noticed during clinical follow-up. CONCLUSIONS: Further investigation to understand the role of rotavirus as a cause of pancreatitis in children may be helpful.

Pathogenic role of myeloperoxidase in acute pancreatitis.

BACKGROUND: Myeloperoxidase (MPO) has been implicated in promoting tissue damage in various inflammatory diseases. However, MPO blood levels in relation to the severity of acute pancreatitis (AP) and its time-course have not been studied. The present study aimed to determine the role of MPO in AP. METHODS: We studied 86 patients with AP (48 patients with mild and 38 with severe pancreatitis) and 18 controls (volunteers). The relations of serum MPO levels to cytokine level, severity, and time-course of pancreatitis were studied. The serum level of MPO and cytokines were measured by MPO-EIA and cytokines ELISA, respectively. RESULTS: The highest level of MPO was noted at the first day in patients with severe AP. A decrease of MPO blood level occurred during the first three days in all patients with necrotizing pancreatitis. The development of pancreatitis-associated lung injury and purulent complications was accompanied by increased MPO levels. Administration of pentoxifylline significantly reduced the MPO blood level, which was clearly correlated with the levels of proinflammatory cytokines in the two groups of patients. CONCLUSIONS: The results of the present study showed the MPO blood level is dependent on the severity of AP and on cytokine blood levels. Pentoxifylline in the complex management of severe AP may improve the results of treatment.