Selected biochemical parameters and ultrastructural picture of pancreas due to Ulinastatin treatment of experimental acute pancreatitis.

Urinary trypsin inhibitor (UTI, Ulinastatin) is a protease inhibitor that has not been yet used in Europe in any experimental trial of severe acute pancreatitis. We have combined the experimental model of severe, hemorrhagic form of acute pancreatitis, and pharmacological treatment with a protease inhibitor. Male Wistar rats were divided into four experimental groups: healthy controls, operated, operated with experimentally induced acute pancreatitis, and animals with acute pancreatitis--treated with UTI preparations. Subjects in the last group were administered UTI intraperitoneally 1 h after pancreatitis induction in an average standard dose of 3000 units/animal. Additionally, four subgroups were created in this treated group, based on the UTI administration time--number of standard doses received: 2 h - 1 standard dose, 6 h - 5 standard doses, 12 h - 11 doses, 24 and 48 h - 15 doses. Statistically significant differences in the serum amylase and lipase activity between the UTI-treated and non-treated subjects were found. In the group of non-treated animals, there a profound destruction of cellular organelles was observed with a total degradation of nuceli, endoplasmatic reticulum and zymogen granules. However, in the UTI-treated subjects, pathological processes proceeded with the significantly slower pace and in much smaller quantities.

Parathyroid adenoma in a subject with familial hypocalciuric hypercalcemia: coincidence or causality?

A middle-aged woman presented with a history of constipation, easy fatigue, depressive mood, lassitude, polydipsia, and polyuria. The patient posed a challenging diagnostic dilemma due to the presence of persistent severe hypercalcemia and relative lack of clinically manifested symptoms. Clinical, biochemical, and genetic examinations confirmed the diagnosis of familial hypocalciuric hypercalcemia as a result of C562Y calcium-sensing receptor mutation, and a coexisting parathyroid adenoma. After adenectomy, the patient's clinical situation improved markedly, and a modest equilibrium hypercalcemia persisted. This case presents an unusual combination of two relatively common endocrine disorders.

Changes in glucose, cholesterol and serum lipid fraction levels in experimental diabetes.

The objective of this research was to trace the changes in serum glucose, cholesterol and lipid fractions during progression of diabetes in rabbits. 89 male rabbits, New Zealand breed were used in the experiment. Diabetes mellitus was induced by a single injection of alloxan. On day 7 the glucose level in the whole blood was measured by a glucometer to confirm the presence of diabetes. From this day the time of disease was counted. The rabbits were divided into the following groups: Group 1--controls (n = 18), Group 2-21 days diabetes mellitus (n = 18), Group 3-42 days diabetes mellitus (n = 17), group 4-90 days diabetes mellitus (n = 19), group 5-180 days diabetes mellitus (n = 17). After above-mentioned periods blood samples were taken and the rabbits were killed by decapitation. The final level of glucose in the sera was determined spectrophotometrically by enzymatic method. The method of cholesterol measurement was based on oxidation of free cholesterol to cholesterol releasing hydrogen peroxide. Measurement of lipid fractions was based on indirect methods consisting in precipitation of specific lipoprotein fractions. Control rabbits revealed highly significant (p < 0.01) or significant (p < 0.05) correlation between initial and final cholesterol and glucose levels. It was not observed in diabetic rabbits. Highly significant correlation (p < 0.01) was found between LDL and total cholesterol concentration in 21 and 42 day of diabetes. Similar correlation was observed between HDL and total cholesterol concentration on 90th day of the course of disease. We concluded that significant disorders of lipid metabolism occur in the course of alloxan-induced diabetes in rabbits, manifested by total cholesterol level increase and changes in proportions and levels of serum lipid fractions.

Lipid and exocrine pancreatic ultrastructural changes due to experimental diabetes.

The aim of this study was to establish if the changes in the ultrastructure of the exocrine part of the pancreas are correlated with changes in serum glucose, cholesterol and lipoprotein fractions during the progression of diabetes in rabbits. Diabetes mellitus was induced in male New Zealand rabbits by a single injection of alloxan into the auricular vein. On the day 7th the glucose level in the whole blood was measured and this day was designated as the first day of diabetes. Rabbits were divided into 5 groups: untreated control, 21-day diabetes, 42-day diabetes, 90-day diabetes and 180-day diabetes. The cholesterol, HDL (high-density lipoprotein) and LDL (low-density lipoprotein) levels were examined in the serum. The total pancreatic lipase activity was measured spectrophotometrically in the pancreatic homogenate. Histological specimens were examined under an electron microscopy. The glucose level increased significantly in all of the alloxan exposed animals. The significant elevation of cholesterol level was observed on day 21 and 180. The HDL level was increased (P<0.05) only on the day 21st. The LDL level and the total activity of pancreatic lysosomal lipase increased significantly on day 21, 42 and 90. Further dilation of granular endoplasmic reticular ducts and decrease in the number of zymogen granules were observed amongst exocrine cells. Fragmented mitochondrial and translucent matrix were also seen. Intensification of the pancreatic fibrosis was found on day 90. Microvascular changes were reported in exocrine cells after 180 days. Their nuclei were smaller with large bulges on the nuclear membrane, and the number of heterogeneous electron granules of zymogen further declined. We concluded that the intensification of ultrastructural changes of the exocrine part of the pancreas correlated with the changes of the pancreatic lipase activity, and glucose and lipoprotein levels.

Changes in pancreatic lysosomal enzymes activity as the potential factors leading to diabetic enteropathy.

The aim of this study was to establish and quantify changes in the activities of total, free and bound fractions of pancreatic lipase, galactoso-6-sulphatase, beta-D-galactosidase and N-acetyl-beta-D-glucosaminidase in the course of alloxan-induced diabetes mellitus. Rabbits were divided into a control group and groups injected with alloxan on the 21st, 42nd, 90th and the 180th day, after which blood samples were taken and the rabbits sacrificed by decapitation. The pancreas was removed and the glucose level measured. Enzyme activities were assayed by spectrophotometric methods. The total activities of N-acetyl-beta-D-glucosaminidase and beta-D-galactosidase were the lowest on day 42 of the test, and the total activity of lipase was the highest at this point of time, as compared to the other periods of the study. We conclude that in the course of alloxan-induce diabetes activities of pancreatic lipase and sulphatase were increasing following the levels of glucose, whilst activities of beta-D-galactosidase and N-acetyl-beta-D-glucosaminidase were declining, being inversely correlated to the level of glucose and activities of the first two mentioned enzymes. Above alterations in activity of lysosomal pancreatic enzymes of alloxan induced diabetic rabbits may be responsible for some aspects of previously reported diabetic enteropathy and chronic complications, or may provide a mechanism for the pancreatic beta-cells to moderate their insulin content.

Temporary elevation of pancreatic lysosomal enzymes, as a result of the omeprazole-induced peripancreatic inflammation in male Wistar rats.

Omeprazole is one of the substituted benzimidazoles, which is not free of side effects. The aim of the study was to evaluate the influence of omeprazole therapy on pancreas. Omeprazole was administered intraperitoneally, twice a day, for 3 days to the male rats in 0.571 mg/kg b.w. and 5.71 mg/kg b.w. doses. Half of animals were sacrificed in the 4th day of the experiment. The remaining rats were raised for another 6 weeks, without any xenobiotics, and sacrificed on the 47th day. The activity of acid phosphatase, beta-galactosidase, cathepsin B, and L, lipase, N-acetyl-glucosaminidase, and sulphatase was evaluated. The slides of the pancreas were examined in light microcopy in hematoxylin-eosin, asan, periodic acid-Schiff (paS) stains. Statistical increase in total activities of acid phosphatase, beta-galactosidase, lipase, N-acetyl-glucosaminidase, sulphatase, and acute inflammatory infiltration in peripancreatic fat tissue without histological pancreas impairment, were observed after the higher dose on the 4th day of experiment. Histological picture and enzymatic profiles were normalized during the next 6 weeks. We concluded that intraperitoneal administration of omeprazole causes tissue inflammation in the peripancreatic lipid tissue and reactive elevation of some pancreatic lysosomal enzymes.

Activity of cathepsins in rat's spleen due to experimentally induced pancreatitis.

The aim of this study was to establish and quantify the changes of the level of cathepsin B, D and L in the spleen during experimental pancreatitis. The experiment was carried out in 115 male Wistar rats, randomly divided into three groups: intact (n = 15), injected with 0.9% NaCl solution into the common bile pancreatic duct (n = 50) and injected with 5% sodium taurocholate into this duct to induce acute pancreatitis (n = 50). After 2, 6, 12, 24 and 48 hours rats were anaesthetised, and blood was taken for amylase determination from the heart, and the spleen was removed. Alpha-amylase level in the blood serum samples was measured by enzymatic method. Cathepsin activity was established by spectrophotometric methods using substrates which form coloured complexes when they react with these proteases. The specific free fraction activity of cathepsin B, D and L in the spleen changed during the course of experiment, but there was no correlation between their activity and the intensity of pancreatitis established by serum amylase level.