Prevalence of diabetes mellitus secondary to pancreatic diseases (type 3c).

BACKGROUND: Diabetes mellitus secondary to pancreatic diseases is a condition seldom thought of in clinical practice. Yet, a high percentage of exocrine pancreatic insufficiency has been reported for the general population and especially for diabetic subjects. Thus, we investigated the prevalence of diabetes mellitus due to pancreatic diseases. METHODS: In this study, we investigated 1868 patients diagnosed with diabetes mellitus who had been admitted to our hospital during the last 24 months. Patient data were diligently studied, and patients were reclassified according to the diabetes classification as proposed by the American Diabetes Association. RESULTS: Among 1868 subjects, 172 patients could be classified as type 3c diabetes mellitus (9.2%). Among these were 135 diagnosed with chronic pancreatitis (78.5%), 12 with hereditary haemochromatosis, 14 with pancreatic cancer and 7 with cystic fibrosis. Thus, diabetes mellitus due to chronic pancreatitis occurred in this collective in 7.2% of all diabetic subjects. Misclassification of these patients was very common. Only 51.2% (88/172) were initially classified correctly. Most type 3 diabetes patients were initially misclassified as type 2 diabetes (69/84). CONCLUSIONS: Diabetes mellitus secondary to pancreatic diseases (especially chronic pancreatitis) seems more common than generally believed with a prevalence of 9.2% among the subjects studied here. Because the awareness of this diabetes type is poor, misclassification is quite frequent. A common problem seems to be the differentiation between type 2 and type 3. Yet, the right classification of diabetes mellitus is important, because there are special therapeutic options and problems in patients with diabetes secondary to pancreatic diseases.

Determinants of Exocrine Pancreatic Function as Measured by Fecal Elastase-1 Concentrations (FEC) in Patients with Diabetes mellitus.

OBJECTIVE: Recently it has been shown that there is not only endocrine insufficiency in diabetic patients, but a frequent co-morbidity of both, the endocrine and exocrine pancreas. The present study was performed to further analyse the determinants of exocrine pancreatic function in patients with diabetes mellitus. METHODS: The records of 1992 patients with diabetes mellitus who had been treated in our hospital during a 2-year period were re-evaluated. Defined parameters were documented in standardized data sheets. Records were further checked for the results of imaging procedures of the pancreas. In 307 patients FEC had been performed and documented. Only these patients were included in further evaluation. RESULTS: FEC was inversely correlated with diabetes duration and HbA1c-levels but not with age. C-peptide levels correlated positively with FEC. BMI and FEC were also significantly correlated. There was no correlation between diabetes therapy and exocrine pancreatic function as there was no correlation with any concomitant medication. The presence of diabetes-associated antibodies was not related to FEC. According to the documented data 38 were classified as type-1 diabetes (12.4%), 167 as type-2 (54.4%), and 88 patients met the diagnostic criteria of type-3 (28.7%). Fourteen patients could not be classified because of lacking information (4.6%). CONCLUSIONS: Exocrine insufficiency might be explained as a complication of diabetes mellitus. However, it is more likely that type-3 diabetes is much more frequent than previously believed. Consequently the evaluation of exocrine function and morphology should be included into the clinical workup of any diabetic patient at least at the time of manifestation.

Modulation of lipoprotein lipase activity by apolipoproteins. Effect of apolipoprotein C-III.

From a total of 22 hypertriglyceridemic subjects tested, 14 subjects were selected on the basis of normal postheparin plasma lipoprotein lipase (LPL) levels and the presence of LPL inhibitory activity in their fasting plasma. The inhibitory activity was detected in both the lipoprotein fraction (d less than 1.25 g/ml) and the lipoprotein-deficient fraction (d greater than 1.25 g/ml). Correlational analyses of LPL inhibitory activity and apolipoprotein levels present in the lipoprotein fraction (d less than 1.25 g/ml) indicated that only apolipoprotein C-III (ApoC-III) was significantly correlated (r = 0.602, P less than 0.05) with the inhibition activity of the lipoprotein fraction. Furthermore, it was found that LPL-inhibitory activities of the plasma lipoprotein fraction and lipoprotein-deficient fraction were also correlated (r = 0.745, P less than 0.005), though the activity in the lipoprotein-deficient plasma was not related to the ApoC-III or apolipoprotein E levels. Additional correlational analyses indicated that the LPL levels in the postheparin plasma of these subjects were inversely related to the levels of plasma apolipoproteins C-II, C-III, and E. To explain some of these observations, we directly examined the in vitro effect of ApoC-III on LPL activity. The addition of ApoC-III-2 resulted in a decreased rate of lipolysis of human very low density lipoproteins by LPL. Kinetic analyses indicated that ApoC-III-2 was a noncompetitive inhibitor of LPL suggesting a direct interaction of the inhibitor with LPL. Results of these studies suggest that ApoC-III may represent a physiologic modulator of LPL activity levels and that the incidence of LPL inhibitory activity in the plasma of hypertriglyceridemic subjects is more common than previously recognized.

Kinetic properties of human pancreatic carboxylesterase.

Fractionation of pancreatic juice by heparin-Sepharose and cholate-Sepharose affinity chromatography indicated that pancreatic carboxylesterase can be separated from pancreatic lipase with the former retained and the latter unretained by both columns. The chromatographic behavior of pancreatic carboxylesterase was found to be similar to that of human milk bile salt-activated lipase. The partially purified pancreatic carboxylesterase had a specific activity of 30 mumol/min per mg protein when assayed with p-nitrophenyl acetate. The reaction mechanism of human pancreatic carboxylesterase was studied using p-nitrophenyl acetate as substrate and taurocholate as activator. The reaction of the enzyme was found to follow a rapid-equilibrium random mechanism. Because of the presence of basal activity, the role of taurocholate can be considered as a non-essential activator and the dissociation constant for the enzyme-taurocholate binary complex was determined to be 0.20 mM. The activation effect of taurocholate consists in increasing the affinity of the enzyme to the substrate (5.6-fold) and in increasing the Vmax (2.3-fold). Based on the kinetic property of human pancreatic carboxylesterase and human milk bile salt-activated lipase with p-nitrophenyl acetate, cholesterol oleate and triolein as substrate, we conclude that they share common substrate specificity but show minor differences in kinetic parameters. Fluorescence studies indicated that both enzymes showed a decreased intrinsic tryptophanyl fluorescence upon incubation with taurocholate. This indicates that bile salt caused a conformational change of the enzymes, with a resultant decreased hydrophobicity in the microenvironment of tryptophan residues.

Fish oil-enriched diet and reduction of low-dose streptozocin-induced hyperglycemia. Inhibition of macrophage activation.

Repeated low doses of streptozocin (STZ; 40 mg/kg, 5 injections/day) induce hyperglycemia in certain strains of mice after a latency of 1 wk. Omega-3 polyunsaturated fatty acids (omega 3FA) have been reported to suppress immune processes by blockade of the cyclooxygenase pathway of arachidonic acid metabolism. We investigated the effects of diets high in omega 3FA on the development of diabetes in the low-dose STZ-induced diabetes (LDSTZ-D) model. Male C57BL/6J mice were on a fish oil diet (FOD) as a source of omega 3FA 8 wk before STZ injection. Controls received laboratory chow only or a coconut oil diet (COD). Blood glucose levels in FOD mice were reduced (12.5 vs. 28 mM for COD mice, P less than .001) 60 days after STZ injection with a diet in which 20% of the calories were from fish oil. In FOD mice, immunohistology showed reduced numbers of class II antigen-expressing cells in pancreatic islets followed by a decreased extent of insulitis. FOD significantly decreased the number of Fc receptor-negative dendritic cells in cytospin preparations of islets isolated from diabetic mice. Interleukin 1-like activity of peritoneal exudate cell supernatants isolated from mice on FOD was reduced. FOD did not improve insulin secretion of isolated islets from LDSTZ-D mice. These data indicate a beneficial effect of FOD on the immune component of the mouse LDSTZ-D model.

Evaluation of serum levels of solubilized adhesion molecules and cytokine receptors in coronary heart disease.

OBJECTIVES: The diagnostic importance of circulating solubilized tumor necrosis factor-alpha receptor II (sTNF-alphaRII) and interleukin-2 receptor in coronary heart disease (CHD) was evaluated. In addition, these variables were correlated with solubilized adhesion molecule levels (i.e., intracellular adhesion molecule [ICAM], vascular cell adhesion molecule [VCAM], and E-selectin). BACKGROUND: Atherosclerosis is considered to be a chronic inflammatory process. Because the immunologic network presents an abundance of positive and negative feedback mechanisms, information obtained from different immunologic variables might be highly redundant. METHODS: In a cross-sectional study design, 60 patients with angiographically proven CHD were compared with 60 individuals who had undergone coronary angiography but in whom no evidence of stenosis could be found (control subjects). Angiography was performed at least one year before the study. Cytokine levels were determined by enzyme-linked immunosorbent assay technique and evaluated by univariate and multivariate statistical methods. RESULTS: All immunologic variables except E-selectin (p = 0.08) significantly discriminated between patients and control subjects. Coronary artery bypass graft surgery after angiography did not lead to significant differences in the variables investigated in the patients with bypass surgery as compared with the subjects without bypass surgery. Receiver-operating characteristics analysis showed comparable test accuracy for solubilized adhesion molecules and cytokine receptors. Multivariate logistic regression analysis, including age, revealed that only ICAM and sTNF-alphaRII were independently correlated with the presence of CHD. Patients belonging to the third tertile of at least one of these two variables demonstrated a 1.6- to 2-fold increased relative risk for the presence of CHD. CONCLUSIONS: We concluded that both circulating ICAM and TNF-alphaRII should be further evaluated as markers for atherosclerotic changes in the coronary system.

Prevalence of risk factors of coronary heart disease in Turks living in Germany: The Giessen Study.

Turkish people represent the majority of immigrants in Germany. Even though a high proportion of Turks has been living in Germany since about 20 years, little is known about risk factors of coronary heart disease (CHD) in this population. In this study a sample of 325 male and 155 female Turks are investigated, who voluntarily underwent a health check-up in Germany. Data about the presence of CHD, risk factors and blood parameters were collected. Mean residence time was 21 and 17 years (males/females). A low percentage of female participants was observed compared to the general Turkish population in Germany. Age adjusted prevalence of CHD reached 9.5% in males and 6.7% in females, respectively. Dyslipoproteinemia (DLP) showed the highest prevalence of all risk factors investigated in both genders. Total cholesterol (TC) levels were comparable to those of other western countries and remarkably higher than reported for the population in Turkey. Besides this, low high density lipoprotein-cholesterol (HDL-C) and apolipoprotein A-I (ApoA-I) levels could be found in the majority of the sample. The highest odds ratios for CHD were estimated for stress and hypertension in males and obesity in females. It is concluded that Turkish immigrants in Germany showed an assimilation of lipid pattern to western populations. However, reasons for low HDL-C levels remain unclear. Changes in the lipid metabolism chiefly seem to contribute to the risk factor pattern of Turkish immigrants in Germany.

Structure and biochemical effects of fenofibrate.

The structures of various fibric acid derivatives are compared. Fenofibrate inhibits de novo hepatic fatty acid synthesis and seems to inhibit hepatic very low-density lipoprotein synthesis, but it enhances mitochondrial and peroxisomal fatty acid oxidation and lipoprotein lipase activity. It produces a very significant reduction in the plasma triglyceride concentration. Fenofibrate also inhibits cholesterol synthesis prior to processing mevalonate, indirectly causing significant reduction of hydroxymethylglutaryl coenzyme A reductase activity. The drug may inhibit acyl-coenzyme A-cholesterol transferase activity, reducing cholesterol ester accumulation within cells. Fenofibrate significantly increases the fractional rate of lecithin-cholesterol acyltransferase activity in normolipidemic and hypercholesterolemic patients. This may explain the increase in cholesterol ester levels observed in high-density lipoproteins. It may stimulate bile acid synthesis from exogenous cholesterol. It causes a marked reduction of increased spontaneous platelet aggregation. Fenofibrate also markedly diminishes the effect of platelet-derived growth factor upon DNA synthesis in vitro, an effect that might impede a key event in early atherogenesis. Thus, fenofibrate has effects not directly related to its lipid- and lipoprotein-lowering action.

Effect of aging on pancreatic lipolytic enzymes.

Pancreatic lipolytic enzyme activities and plasma lipids were measured in three age groups of female Fisher 344 rats (3 months [young], 12 months [adult], and 27 months [old]) in order to evaluate age-related changes and a possible correlation of these parameters. Cholesterol esterase activity was measured in pancreas homogenate, while the lipase activity was further fractionated into heparin-Sepharose unretained (lipase I) and retained (lipase II) fractions. In analogy to the lipolytic enzymes of the human pancreas, lipase I corresponds to pancreatic lipase and lipase II corresponds to pancreatic carboxylesterase. Plasma triglyceride and cholesterol levels were significantly higher in old as compared with adult and young rats. There was no significant correlation between plasma lipid and pancreatic enzyme activity levels. Cholesterol esterase and pancreatic lipase (lipase I) activity did not show any consistent change with age. Pancreatic carboxylesterase (lipase II), on the other hand, was consistently lower in adult and old animals. Although the importance of pancreatic carboxylesterase for triglyceride hydrolysis remains to be established, our results suggest that this enzyme is under long-term metabolic control. We suggest that early in life, high levels of this enzyme may be needed to supplement pancreatic lipase in order to optimize digestion of dietary fat required for optimal growth.

Tumor M2-pyruvate kinase: a promising tumor marker in the diagnosis of gastro-intestinal cancer.

The diagnostic value of pyruvate-kinase type tumor M2 (Tumor M2-PK) has been investigated in different tumors, and showed interesting results in cases of renal cancer, pancreatic cancer, lung cancer and some cases of gastric cancer. In this study we investigated EDTA-plasma of 68 patients with gastrointestinal cancer, 22 patients with inflammatory bowel disease (IBD) and 60 healthy controls. Sensitivity of Tumor M2-PK was 70.6% for all GI-tumors, that of CA19-9 was 55.4% and that of CEA was 53.3%. In pancreatic cancer CA19-9 showed the best sensitivity. In oesophageal/gastric cancer Tumor M2-PK was most sensitive and in colorectal cancer CEA and Tumor M2-PK showed the best results. The specificity of Tumor M2-PK was 90-96, 7%. In IBD some individuals showed elevated Tumor M2-PK levels but there was no correlation to CRP or to the clinical activity score. The results indicated that Tumor M2-PK might be a valuable marker in gastrointestinal cancer.

Measurement of fecal pyruvate kinase type M2 (tumor M2-PK) concentrations in patients with gastric cancer, colorectal cancer, colorectal adenomas and controls.

BACKGROUND: The determination of Tumor M2 Pyruvate Kinase (Tumor M2-PK) in EDTA plasma has been described as a tumor marker in a variety of different tumor types. Since most GI tumors grow intraluminally, it appeared interesting to determine whether Tumor M2-PK is detectable in the feces of tumor patients. MATERIALS AND METHODS: Measurements were performed with a commercially available ELISA (ScheBo Biotech AG, Giessen, Germany) modified for fecal analysis. Samples of controls, GI cancer and adenoma patients were tested. RESULTS: Fecal Tumor M2-PK concentrations could be quantified. A significant difference between cancer patients and controls was found. The highest concentrations were observed in colorectal cancer. However, fecal Tumor M2-PK was not detectable in 5 tumor patients. CONCLUSION: The measurement of fecal Tumor M2-PK concentrations might provide an interesting screening tool for colorectal cancer. Further studies comparing the determination of Tumor M2-PK in stool to occult blood in stool are in progress.

Fecal pyruvate kinase concentrations (ELISA based on a combination of clone 1 and clone 3 antibodies) for gastric cancer screening.

BACKGROUND: Recently we were able to demonstrate that Tumor M2 Pyruvate Kinase (Tumor M2-PK) is detectable in the feces of patients with gastrointestinal (GI) cancer. In this study an ELISA based on a different combination of antibodies was used to investigate stool samples of GI cancer patients. PATIENTS AND METHODS: The ELISA (ScheBoBiotech AG, Giessen, Germany) was based on one antibody specific for Tumor M2-PK and a second antibody reacting with Tumor M2-PK, M1-PK and the tetrameric M2-PK. Stool samples of healthy controls, patients with inflammatory bowel disease (IBD) and different kinds of GI cancer were tested. RESULTS: Compared to controls, samples of IBD or different tumors did not show significant differences, except those patients with gastric cancer (elevated fecal PK in 80% (p = 0.005)). CONCLUSION: If the present results can be confirmed in larger patient samples, this new test might provide an excellent screening tool for gastric cancer.

Influence of cholesterol and fat in food on indirect parameters of exocrine pancreatic function.

Fecal elastase 1, chymotrypsin activity, and fat content in stool are clinical parameters of exocrine pancreatic function. The aim of this study was to clarify the possible impact of extreme changes in diet on fecal enzyme concentration/activity, since extreme diets may lead to wrong conclusions in the diagnosis of pancreatic insufficiency. Twelve healthy test persons followed 4 diet phases, each with a duration of 6 days. The 4 diet phases were (1) nearly fat-free with a low-cholesterol content; (2) high in fat and low in cholesterol; (3) high in cholesterol deriving from meat, and (4) high in cholesterol deriving from eggs. At the end of each diet phase, a 72-hour stool collection was carried out to measure fecal elastase 1, chymotrypsin and fecal fat content. The results showed no significant changes after each of the 4 diet phases. The clinical parameters of fecal elastase 1 and chymotrypsin activity in stool do not seem to be significantly influenced by fat and cholesterol deriving from food.

[Pyruvate kinase M2 (tumor M2-PK) as a screening tool for colorectal cancer (CRC). A review of current published data]. / Pyruvatkinase M2 (Tumor M2-PK) im Stuhl als Screeningparameter für kolorektale Neoplasien. Eine Ubersicht über bisher publizierte Daten.

INTRODUCTION: Colonoscopy is currently supposed to be the best screening tool for colorectal cancer. However, the acceptance of the population is very poor although it has been included in screening programs in the German health system since 2002. Therefore, evaluation of additional screening tools seems to be of great interest. Recently testing for fecal occult blood (FOBT), genetic alterations or alterations in tumor metabolism (e.g., tumor M2-PK) are under investigation. METHODS: The use of M2-PK measurement in the feces has been reported in 6 studies until today. The data of these studies were analyzed and critically reviewed. RESULTS: The overall sensitivity of M2-PK is 77.9% concerning CRC. Specificity ranges from 74.3-83.3%. Overall sensitivity for adenomas is 45.9%, increasing to 61.1% for adenomas > 1 cm. A high percentage of positive results (90.4%) was also observed in patients with chronic inflammatory bowel disease. CONCLUSIONS: Compared to FOBT or genetic testing the M2-PK test seems to be superior for CRC screening. Concerning handling, effectiveness and analysis, M2-PK seems to be a good possibility for large scale-screening of colorectal carcinoma. It might even be used to detect larger adenomas. Elevated levels of M2-PK in patients with acute and/or chronic inflammatory bowel diseases are probably due to proliferation of epithelial cells and leucocytes in the inflammatory area.

Purification of human lysozyme from milk and pancreatic juice.

Human milk lysozyme was purified by heparin-Sepharose affinity chromatography and Sepharose 4B gel-permeation chromatography. This procedure was also found applicable to the purification of human pancreatic juice lysozyme. Double-diffusion analyses indicated that human milk lysozyme was immunochemically identical to human saliva and human pancreatic juice lysozyme. Based on the identity of the N-terminal 10-amino-acid-residue sequence analyzed, it was suggested that human milk lysozyme and human pancreatic juice lysozyme are identical molecular entities.

Determination of apolipoprotein variants by isoelectric focusing in agarose.

Isoelectric focusing (IEF) of apolipoproteins is an important procedure for the isolation of apolipoprotein isoforms. In this paper we describe the use of agarose/urea as a focusing medium for IEF of apolipoproteins. This technique, which offers several advantages over the traditional polyacrylamide gel focusing, yields a further microheterogeneity of the known isoforms of apolipoproteins A-I and E. In addition, by subsequent immunoprecipitation, both specificity and sensitivity are enhanced. Using a monospecific antiserum against Apo A-I, a new minor isoform was detected at the basic end of the Apo A-I spectrum (pI 6.02) which has not yet been observed in normal plasma samples. Another advantage of this technique in combination with immunoprecipitation is the specific detection of Apo E phenotypes which is essential for the diagnosis of hyperlipoproteinemia type III.

CI-924 effects on plasma lipids in patients with type II and type IV hyperlipoproteinaemia.

CI-924 (CI), 5,5'-[[1,1'-biphenyl]-2,5-diylbis(oxy)]bis[2,2- dimethylpentanoic acid] is chemically similar to gemfibrozil. Patients with Type II (n = 13) and Type IV (n = 22) hyperlipoproteinaemia (HLP) were maintained 12 weeks on a baseline diet containing 55% sugar, 15% protein 30% fat and less than 300 mg cholesterol daily to stabilize weight and lipids. They were then entered in a parallel group double-blinded protocol and received 0, 300, 600, or 1200 mg CI p.o. daily for 12 weeks. CI consistently elevated anti-atherogenic HDL and lowered VLDL at 600 mg/day in both Type II and Type IV HPL at 8 weeks. In Type II patients, CI lowered cholesterol, decreased LDL/HDL and increased ApoA-I. In Type IV patients, CI also lowered TC while elevating LDL and ApoA-II. CI had no effect on Apo-B, LDL-ApoB, or Apo-E.

Comparative studies on the chemical and immunochemical properties of human milk, human pancreatic juice and bovine milk lactoferrin.

Lactoferrin from human milk, pancreatic juice and bovine milk were purified by heparin-Sepharose affinity chromatography procedure. Urea-sodium dodecylsulfate-polyacrylamide gel electrophoresis of these lactoferrin preparations indicated similar molecular weights (80,000). Metal analyses showed that lactoferrin of bovine milk contained the highest amount of iron while lactoferrin of human milk and human pancreatic juice were similar in content of iron, approximately four-fold lower than bovine milk. All these lactoferrin preparations were also found to contain minor amounts of copper and manganese. Double diffusion analyses indicated that lactoferrin of human milk was immunochemically identical to lactoferrin of human pancreatic juice. On the other hand, immunochemically, bovine milk lactoferrin was not identical to human milk lactoferrin. Sequence analyses of human milk and pancreatic juice lactoferrin indicated that they shared the same N-terminal sequence for the 16 residues analyzed. Although human milk and bovine milk lactoferrin had sequence homologies, bovine milk lactoferrin had a closer homology to ovotransferrin than to human milk lactoferrin.