Complications of endoscopic retrograde cholangiopancreatography (ERCP) and their risk factors.

PURPOSE: Determination of the type and frequency of complications developing after diagnostic and therapeutic endoscopic retrograde cholangiopancreatography (ERCP) as well as the risk factors predisposing to them. MATERIAL/METHODS: The retrospective study, including 734 ERCP performed in 550 patients, with 404 (55%) ES (endoscopic sphincterotomy) during a 4-year period. RESULTS: Among 734 ERCP procedures, 76.4% (561) had both diagnostic and therapeutic purpose, 15.2% (112) were only diagnostic. Complications developed after 26 procedures (3.5%): acute pancreatitis (AP) in 8 patients (1.09%), cholangitis in 7 (0.95%) and delayed bleeding in 11 (1.5%) patients. After 49 (6.7%) ES immediate bleeding was observed. The risk factors for AP were: unintentional pancreatic duct contrasting, mechanical lithotripsy, the use of the "pre-cut" technique and bile duct dilatation. Cholangitis was more common in cases with difficult cannulation at older age and with lower baseline bilirubin level. The risk factors for delayed bleeding were: location of the ampulla of Vater in the diverticulum and the use of the "precut" technique. Immediate bleeding was more frequent after revision of bile ducts with Dormia's basket or with balloon, after introduction of contrast medium to the pancreatic duct or in ductal cholelithiasis. CONCLUSIONS: ERCP performed in the endoscopy unit of a specialist hospital department is a relatively safe procedure, with a low burden of complications as compared to the benefits it provides to appropriately qualified patients.

Exocrine pancreatic function in biliary tract pathology treated with the endoscopic methods.

PURPOSE: Incidence of pancreatic exocrine insufficiency in biliary pathology is estimated for about 30%. The objective was to assess pancreatic exocrine function in biliary tract pathology (cholelithiasis, strictures) before and after endoscopic treatment. PATIENTS AND METHODS: Twenty-eight patients with choledocholithiasis and its complications (19F/9M; aging 31-90 years, median: 69 years) were evaluated. Fecal elastase 1 concentration was measured using ELISA, before, early, and 6-8 weeks after endoscopic treatment. The inflammatory response of pancreas to the treatment was also assessed. RESULTS: Initial fecal elastase 1 concentration in patients (median 454 microg/g) was not significantly different as compared to the control (median 357 microg/g). Nine patients (32%) had low fecal elastase 1 concentration (below 250 microg/g) and out of them 6 had the concentration below 200 microg/g, suggesting impairment of exocrine pancreatic function. Endoscopic treatment was successful in 82% of patients. Pancreatic inflammatory response was noted only in one patient. After 6-8 weeks fecal elastase 1 concentration in the whole group of patients did not significantly change in comparison to the initial level. However, out of 9 patients with initially low fecal elastase 1 concentration (median 191 microg/g) at least in 6 pancreatic function improved (median 310 microg/g), P < 0.001. CONCLUSION: One third of the patients with biliary pathology had a low fecal elastase 1 concentrations, suggesting pancreatic dysfunction. In at least 2/3 of these patients successful endoscopic treatment of biliary pathology resulted in the significant increase of fecal elastase 1 concentration. Therefore, an additional positive effect of such treatment in some patients, could be an improvement of the exocrine pancreatic dysfunction.

The relationship between masticatory efficiency and the state of dentition at patients with non rehabilitated partial lost of teeth.

UNLABELLED: Factors believed to affect masticatory efficiency include loss of postcanine teeth, bite force, severity of malocclusion, occlusal contact area, body size and oral motor function. THE AIM: to record if there is relationship between masticatory efficiency and the state of dentition at patients whose occlusion has never been rehabilitated. MATERIAL: The study was performed in 22 patients who were missing over 50% of their functional dental units and never used any prosthetic appliances and in 15 healthy completely dentate controls. METHODS: The masticatory efficiency was measured using Optosil test for 20 and 80 cycles of chewing. The occlusal conditions were analyzed by means of the computerized T-Scan II System which registered the maximal force of pressure during the maximal occlusal contacts, the time which passed between the first contact and the maximal force of pressure and the occlusal platform area. RESULTS: It was observed a considerable difference in the integrity of the masticatory system between both groups. The force of pressure on the indicator, chewing platform area and the time from the first contact to the maximal force calculated in T-Scan II System differs significantly between both groups. The value of X50 for 20 and 80 cycles of chewing estimated in Optosil test were statistically significant only for 80 cycles of chewing. CONCLUSION: The severe reduction of the number of functional dental units is caused of the impairment of chewing ability but prolongation of mastication could improve the comminution of hard food.

The cumulative effect of nuclear factor-kappaB (NF-kappaB) inhibition and endothelins in early cerulein-induced acute pancreatitis in rats.

PURPOSE: 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. MATERIAL AND METHODS: 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. RESULTS: %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. CONCLUSIONS: 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.

Effects of endothelin-1 or of its receptor A a selective antagonist, on histological and ultrastructural patterns in experimental acute pancreatitis in rats.

The role of endothelin-1 (ET-1) and of its receptor A (ETA) blockade in oedematous acute pancreatitis (AP) remains unclear. In 40 male Wistar rats with i.p. cerulein-induced AP, lasting 4 hours, ET-1 2x0.5 nmol/kg and 2x1.0 nmol/kg or selective ETA antagonist LU 302146, 10 mg/kg and 20 mg/kg was given i.p. simultaneously with cerulein. Histological and ultrastructural studies of pancreatic specimens were done. ET-1 decreased the inflammatory infiltration, but not the mean scores of necrosis and vacuolization in AP. The ultrastructural damage of acinar cells was less evident after ET-1 than in untreated AP. Selective ETA antagonist slightly aggravated the vacuolization and necrosis of acinar cells and some ultrastructural alterations in AP. In conclusion, ET-1, in contrast to selective ETA antagonist, exerts some protective effect in the early course of oedematous cerulein-induced acute pancreatitis in rats.

The effect of endothelin-1, endothelin-2 and endothelin-3 in early cerulein-induced acute pancreatitis in rats.

PURPOSE: 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. MATERIAL AND METHODS: 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. RESULTS: %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. CONCLUSIONS: 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.

The experimental distention of dissected bile duct for the restoration of its continuity in dogs using a device of own construction.

PURPOSE: The segmental resection of constricted bile duct and end-to-end biliary anastomosis could be an attractive alternative in the treatment of benign biliary tract stricture. The aim of this study was to restore the anatomical integrity of the hepatic-common bile duct after an artificially produced defect while maintaining the large duodenal papilla, using microsurgical technique. MATERIAL AND METHODS: The experiments were carried out on 25 mongrel dogs. The common bile duct was ligated in all of the animals during laparotomy, as a model of bile duct obstruction in humans. Relaparotomy was performed 3 days after the initial operation. The segment of bile duct, 4 cm in length was resected together with the ligature. The continuous bile flow into the duodenum was assured by a polyvinyl catheter introduced into both ends of dissected bile duct. The proximal end of the hepatic-common bile duct was fixed to a device constructed by us for the distention of the bile duct (DDBD). The anterior part of the device was exteriorized through a separate fistula and fixed to the abdominal wall. The hepatic-common bile duct distention was gradually continued during 18 days, by pulling out the mobile part of the device. After 18 days the device was removed and the distended proximal end of the hepatic-common bile duct was anastomosed end-to-end with its distal end. The sequels of this procedure were observed for up to 6 months. RESULTS: The hepatic-common bile duct was distended 4 cm within 18 days. The histopathological examination has shown partial damage of the duct framework due to the distention and tension. However the patency of the duct was preserved and the recovery of normal structures were observed after the device was removed and anastomosis fashioned. CONCLUSION: This method, developed by us, offers the possibility of restoring the integrity of injured extrahepatic bile ducts, allowing effective treatment of benign biliary strictures.

Lysosomal activity of pulmonary alveolar macrophages in acute experimental pancreatitis in rats with reference to positive PAF-antagonist (BN 52021) effect.

The activation of pulmonary alveolar macrophages (PAM's), might play an important role in severe complications of acute pancreatitis. The aim of our study was to assess the labilization of macrophage lysosomal membranes and release of lysosomal cathepsin B (CB) and N-acetyl-beta-D-hexosaminidase (NAH) into bronchoalveolar lavage fluid (BALF) during taurocholate acute pancreatitis (AP) in rats treated with PAF-antagonist--BN 52021. Total activity of CB increased by 374% after 6 h and by 237% after 12 h of AP in lysosomal enriched fraction of PAM's. Fractional free activity of CB increased to 40% after 6 h and to 38% after 12 h of AP. Free activity of CB was increased 5 fold in the supernatant of macrophage homogenate, and 10 fold in the supernatant of BALF after 6 h of AP. The values of NAH activity roughly paralleled that of CB. Treatment with BN 52021 (5 mg x kg(-1) every 6 h i.v.) partially normalized the measured parameters. Our results indicate that the PAF-antagonist BN 52021 reduced the increase of total and free activity of lysosomal hydrolases of PAM's and partly prevented the labilization of their lysosomal membranes. Therefore, an important mechanism of BN 52021 beneficial effect in pulmonary complications of acute pancreatitis could be dependent on the stabilization of PAM's lysosomes.

The effect of antecedent acute ethanol ingestion on the pancreas ultrastructure in taurocholate pancreatitis in rats.

The rat pancreas ultrastructure was examined 6, 12, and 18 h after (1) taurocholate-induced acute pancreatitis and after (2) pancreatitis preceded 6 h earlier by intragastric acute 40% ethanol ingestion (5 g/kg b.w.). Pancreatic specific trypsin activity and plasma alpha-amylase were assayed at the same time intervals. The antecedent acute ethanol ingestion resulted in the evident aggravation of pancreas ultrastructural alterations. Acute pancreatitis preceded by ethanol resulted in the increase of zymogen granules number, RER channels were more irregularly distributed, autophagosomes were more abundant and degeneration of mitochondria was more advanced when compared to acute pancreatitis without ethanol ingestion. Tryptic activity increased to higher degree in all pancreatitis groups preceded by ethanol, but this difference was statistically significant (P < 0.01) only after 18 h. These morphological (but not biochemical) differences progressed 12 h after pancreatitis induction. After 18 h of acute pancreatitis the number of zymogen granules decreased in previously alcoholized rats, but tryptic activity remained twofold higher that in animals not given ethanol. Other signs of cellular impairment were still more prominent in alcoholized rats. The obtained results suggest that even single acute ethanol abuse prior to acute pancreatitis does aggravate the morphological and biochemical lesions observed in this disease with possible negative consequences for the prognosis.

The liver ultrastructure in caerulein and taurocholate acute pancreatitis in the rats.

The purpose of the study was to compare the morphological alterations of the liver in two models of acute pancreatitis: caerulein-induced (edematous) and taurocholate-induced (necro haemorrhagic one). The experiments were performed on 24 male, Wistar rats, weighing 240-260 g. In group I (n = 8) the supramaximal stimulation with i.v. caerulein (5 micrograms/kg/h) during 12 h was applied (C-AP). Control animals (group II, n = 4) received i.v. saline (C-C). In group III (n = 8) 5% sodium taurocholate (0.2 ml/min) was injected into the bile-pancreatic duct during sterile laparotomy (T-AP). In group IV (n = 4) animals were sham operated (T-C). The specimens of the liver were excised after decapitation of rats at 12 h after beginning of caerulein infusion or intraductal injection of sodium taurocholate. The light and electron microscopy was performed. The marked hepatic lesion were found in both variants of experimental pancreatitis, however they were far more advanced in taurocholate pancreatitis. In light microscopy the dispersed foci of colliquative necrosis, degeneration of hepatocytes, swelling of Kupffer cells predominated in taurocholate pancreatitis. The glycogen deposits were depleted but lipid droplets were increased in size and number. The swelling of mitochondria, degeneration of their matrix and cristae, increase of autophagocytosis and numerous lysosomes, the lesions of sinusoids with increased activity of phagocytic cells were more evident in taurocholate pancreatitis--(more severe model of the disease). These findings document severe injury to the liver in acute pancreatitis depending on the severity of inflammatory process in pancreas. They also suggest that the liver could be not only passive target of pancreatogenic noxa in acute pancreatitis, but it could be also a defensive barrier against spreading of injuring agents on other system. This role seems to be especially evident in more severe form--taurocholate induced pancreatitis.

The effect of beta-thia-iminoprostacyclin in taurocholate acute pancreatitis in rats: the role of antecedent acute ethanol abuse.

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.

Does antecedent ethanol intake affect course of taurocholate pancreatitis in rats?

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.

The effect of prostacyclin analogue--iloprost on the pancreas regeneration after caerulein-induced acute pancreatitis in rats.

UNLABELLED: Prostacyclin (PGI2) and its stable analogue iloprost (I) exert beneficial effect in acute pancreatitis (AP). The aim of the study was to evaluate the role of iloprost in pancreas regeneration after AP in rats. METHODS: AP was induced in male Wistar rats by s.c. injections of caerulein 12 micrograms/kg t.i.d. for 2 days. Rats were divided into four groups: control + saline (C); control + iloprost (C/I); AP + saline (AP); AP + I (AP/I). Rats were treated for 7 or 14 days. I (Schering AG) was given at the dose I microgram/kg b.w., i.p., t.i.d. After the rats were killed, the pancreata were weighed and their protein, DNA, RNA, chymotrypsin, alpha-amylase contents were evaluated. Light microscopic examination of representative pieces of pancreas was performed. RESULTS: Acute pancreatitis resulted in pancreas destruction observed even 7 days after the onset of the disease. The significant decrease of pancreatic weight, RNA and chymotrypsin contents were observed in AP rats when compared to C. The improvement of pancreatic histology and significant increase of DNA content were found in I treated (during 7 days) AP rats in comparison to untreated AP group. Two weeks after pancreatitis induction the pancreas regeneration occurred in both pancreatitis groups and it was connected with pancreas hypertrophy. Treatment with I resulted in slight not significant increase of some of cellular hypertrophy indices when compared to AP untreated animals. Healthy rats injected with I during 7 days showed significant elevation of DNA content in comparison to C. When treatment with I was prolonged up to 14 days such hyperplastic effect was not observed. Our results suggest, that treatment with iloprost exerts temporary hyperplastic influence on the pancreas of healthy rats and pancreas regenerating after caerulein-induced pancreatitis.

The effect of tilsuprost on the liver mitochondria in taurocholate pancreatitis in rats with antecedent acute ethanol abuse.

The damage to the liver appears to be an important aspect of multisystem organ failure in acute pancreatitis with poor prognosis. The objective of this study was to evaluate the protective effect of stable prostacyclin analogue--tilsuprost on the liver energy metabolism in taurocholate pancreatitis in rats preceded by acute ethanol intake. The respiratory control ratio (RCR) and ADP/O ratio of liver mitochondria with glutamate+malate as substrates and mitochondrial DNP (uncoupler)-dependent ATPase activity were significantly depressed after 12 h of taurocholate pancreatitis-the effects that were not significantly aggravated by antecedent acute ethanol intake. Tilsuprost (0.3 mg/kg i.g.) given just before induction of pancreatitis partly prevented the impairment of mitochondrial oxidative and phosphorylative functions, however these positive effects were limited in acute pancreatitis preceded by acute ethanol intake. These results suggest that prostacyclin analogues could be effective in the treatment of hepatic complications in acute pancreatitis, however their effectiveness could be limited in the case of acute ethanol antecedent abuse.

Hepatic mitochondrial and lysosomal alterations in acute experimental pancreatitis with ethanolic coetiology in rats.

In order to assess the cumulative effects of antecedent acute ethanol intake and acute pancreatitis on the liver, the mitochondrial respiratory functions and lysosomal membrane integrity of the liver were evaluated in taurocholate pancreatitis (AP) in rats, induced 6 hr after intragastric ethanol 40% (5 g/kg body wt). The oxygen consumption rate, RCR (respiratory control ratio), and ADP/O ratio were measured according to Estabrook. Fractional free activity of lysosomal hydrolases was assayed. RCR with glutamate + malate was most decreased at 12 hr of AP with partial improvement after 18 hr. The ADP/O ratio dropped maximally after 18 hr of AP. The fragility of lysosomal membranes increased significantly at 18 hr of AP. The antecedent ethanol intake abolished the partial restoration of RCR after 18 hr; however, it did not affect the ADP/O ratio or the integrity of lysosomal membranes impaired in AP at this time. In conclusion, the antecedent acute ethanol abuse could aggravate the liver mitochondrial deterioration, but not the lysosomal membrane labilization seen in AP.

The time course of ultrastructural changes in the secretory components of pancreatic acinar cells and trypsinogen activation in taurocholate pancreatitis in rats.

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.

The lysosomal hydrolases in the rat pancreas after maximal or supramaximal stimulation with cerulein.

The decompartmentation of lysosomal compartment in pancreatic acinar cells with consecutive activation of zymogens might play an important role as a "trigger mechanism" in acute pancreatitis. The admixture of lysosomal hydrolases to secretory enzymes in pancreatic juice was found, but their role in pancreatic secretion remains obscure. The aim of the present study was to assess the fragility of pancreatic lysosomal structure after maximal (optimal) or supramaximal stimulation of rats with cerulein during 3, 6, 12 h, and after recovery. In the mitochondrial-lysosomal (M-L) and in the supernatant (S) of pancreases free (F) total (T), and fractional free (%F/T) activities of beta-glucuronidase (beta G), acid phosphatase (AcP), cathepsins (Cs), and beta-N-acetyl-hexosaminidase (NAH) were estimated. In edematous pancreatitis following supramaximal stimulation with cerulein, a significant increase of %F/T of beta G in whole homogenate began at 6 h of hyperstimulation in comparison to the control (93 vs 42% p < 0.01). This increment persisted until 12 h of hyperstimulation and declined after 24 and 48 h of recovery to 67-69%. The changes of %F/T of beta G in M-L followed those in whole homogenate, and additionally the increase free activity in S after 6 h of hyperstimulation and after 24 h recovery occurred. The respective activities of other hydrolases showed a similar pattern of changes. It is of interest that fragility of lysosomal membranes increases significantly also after maximal stimulation when inflammatory changes were absent. Our results suggest that the increase of lysosomal fragility of the pancreas is most unlikely pathological in itself, but also occurs during stimulated pancreatic secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Cathepsin B inhibition in two models of acute pancreatitis.

The possible role of cathepsin B in the pathogenesis of two forms of acute pancreatitis was studied using the cathepsin B inhibitor known as E-64. In an edematous, nonfatal pancreatitis induced by supramaximal doses of cerulein, increases in the serum amylase and lipase levels were less pronounced in rats pretreated with E-64. Other parameters of pancreatic injury were unaffected by inhibition of cathepsin B. In a necrohemorrhagic type of pancreatic injury induced by retrograde infusion of bile salts into the pancreatic duct, E-64 partially attenuated increases in serum levels of amylase and lipase, and in addition, reduced the activation of trypsinogen. However, the high mortality in this model of pancreatitis was not modified.

The role of lysosomal alterations in the damage to the pancreas and liver in acute experimental pancreatitis in dogs.

The role of lysosomal hydrolases in the pathogenesis of acute pancreatitis and secondary liver injury, as an important aspect of multisystem organ failure, remains unclear. The purpose of this study was to assess the lysosomal fragility in both organs in acute experimental pancreatitis (AEP) of graded severity in dogs. In 7 dogs, the moderate (M) and in 13 dogs severe (S) variant of bile--trypsin AEP--was induced; 6 dogs were in control group (C). The 24 h survival time was 6/7 and 6/13, respectively. After that time, the dogs were sacrificed and the lysosomal enriched subfraction (L) from both organs was isolated by ultracentrifugation. The total (T) and free (F) activities of beta-glucuronidase (beta G), cathepsins (Cs) and acid phosphatase (AcP) according to Gianetto and de Duve were assayed. The fractional free activity (% F/T) was adapted as and index of lysosomal stability. The %F/T of BG in the homogenate of the pancreas in AEP(S) was higher than that in AEP(M) (92% vs. 71%, p < 0.05, and vs. 37% in C, p < 0.005). The %F/T of Cs and AcP showed a similar pattern. The %F/T of beta G in L of the liver in AEP(S) was 38% vs. 29% in AEP(M), (p < 0.05), and vs. 20% in C (p < 0.05). In AEP in dogs the %F/T activities of lysosomal hydrolases in the pancreas and liver were increased, suggesting the labilization of lysosomal membranes in severe form of this disease. Our results support the pathogenic role of lysosomal hydrolases in the damage to the pancreas and liver in acute pancreatitis.

Severity of Helicobacter-induced gastric injury correlates with gastric juice ammonia.

We postulated that ammonia produced by Helicobacter pylori may contribute to gastric mucosal injury. This hypothesis was evaluated in Helicobacter-positive patients with chronic renal failure in whom a high urea concentration might amplify this phenomenon. Gastric urea and ammonia were measured, and the severity of gastritis was evaluated by counting mononuclear and polymorphonuclear cells. High gastric ammonia and low urea in Helicobacter-positive patients, and the converse in Helicobacter-negative subjects, were observed. There was a significant correlation between gastric ammonia and interstitial polymorphonuclear leukocytes infiltration (P less than 0.05), suggesting a causal link. Eradication of Helicobacter pylori was associated with a decrease of ammonia and an increase of urea (P less than 0.01). The significant correlation between the severity of gastric inflammation and the gastric juice ammonia concentration suggests that ammonia may play a pathogenic role in Helicobacter-associated gastric injury.