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J F Rehfeld

Publications and source records attributed to J F Rehfeld.

At least 73 records · Page 4Linked to original sources
PubMed1999

CCK stimulates growth of both the pancreas and the liver.

BACKGROUND: The pancreas is the main target of the trophic effect of cholecystokinin (CCK), with a transient increase in cell proliferation and persistent effect on DNA content and weight gain when given continuously. Whether CCK exerts similar effects on the liver and biliary tract is not known. Most studies on this subject have hitherto been done in the rat. The aim of the present experiments was therefore to study the possible concomitant trophic effects of CCK on these three organs in a gallbladder-bearing animal, the hamster. METHODS: CCK-8S or the CCK-A receptor antagonist devazepide were infused subcutaneously by osmotic minipumps for 4 weeks in 11 and 7 hamsters, respectively. Fifteen animals served as untreated controls. One hour before sacrifice tritiated thymidine was injected intraperitoneally. Plasma was collected for determination of CCK and the pancreas, liver, common bile duct and gallbladder were excised. The pancreas and liver were weighed and processed for the contents of protein, DNA and water. Tissue specimens were taken for autoradiography. RESULTS: The concentration of CCK in plasma increased fourfold during the infusion of CCK-8S. The pancreas and, to a lesser extent, the liver increased in weight, DNA and protein contents after infusion of CCK-8S, whereas the pancreas, but not the liver, decreased in weight and protein content after infusion of devazepide. Pancreatic amylase content was increased after CCK-8S treatment, but unaffected by devazepide. Labelling index of pancreatic acinar cells, hepatocytes and gallbladder epithelium was no different from that of controls after four weeks of infusion of CCK-8S or devazepide. A correlation was found between the concentration of plasma CCK on one hand and the weights and DNA contents in pancreas and liver on the other. CONCLUSION: The results suggest that CCK stimulates growth of both the pancreas and the liver in the hamster.

Animals↗
PubMed1998

How to measure cholecystokinin in tissue, plasma and cerebrospinal fluid.

This review examines a major problem for an old hormone. Hormones are defined by the ability to reach their targets via blood. Consequently, knowledge about a hormone requires measurement of its behaviour in blood. So far, however, it has proven exceptionally difficult to measure the classical gut hormone, cholecystokinin (CCK), in circulation. The review therefore describes the premises for reliable plasma CCK measurements as compared to the premises for measurement in tissue extracts and cerebrospinal fluid. The critical plasma premises comprise equimolar quantitation of the bioactive CCK peptides in circulation (CCK-83, -58, -33, -22 and -8) without interference from homologous gastrin peptides. The latter may appear nearly impossible, because the bioactive epitopes of CCK and gastrin are almost identical, and because the plasma concentrations of gastrin are more than tenfold above those of CCK. In comparison, measurement of CCK in tissue is considerably simpler, especially in extracts of the two main production sites, the brain and jejunoileal mucosa. For cerebrospinal fluid, degradation, low levels and shortage of material constitute major problems so that the molecular nature and biological/clinical relevance of CCK measurements in CSF still remain to be settled. The review finally enlists the reports on plasma CCK measurements published so far. A multitude of different immuno- and bioassays have been used with corresponding variation in the results. The theory for different types of assays in combination with general assay experience suggest that accurate CCK measurements require radioimmunoassay technology based on high-affinity antibodies. These antibodies have to be exquisitely specific for the 0-sulfated C-terminal heptapeptide amide of CCK without binding the similar gastrin epitope. Only few of such antibodies have been raised.

Animals↗
PubMed1998

Altered processing of procholecystokinin in carboxypeptidase E-deficient fat mice: differential synthesis in neurons and endocrine cells.

The fat mouse strain exhibits a late-onset obesity syndrome associated with a mutation in the gene encoding carboxypeptidase E (CPE). CPE plays a central role in the biosynthesis of many regulatory peptides. Therefore, we examined the processing of procholecystokinin (proCCK) in the brain (neurons) and small intestine (endocrine cells) of fat/fat mice. In the brain, bioactive CCK was markedly reduced (7.9+/-1.0 pmol/g in fat/fat mice vs. 82.5+/-11.2 pmol/g in controls), but the concentration of the CPE substrate, glycylarginine-extended CCK, was elevated 105-fold. In contrast, the concentration of bioactive CCK in intestinal endocrine cells was unaffected. Endocrine cell processing was, nevertheless, altered with a 33-fold increase in glycyl-arginine-extended CCK. Interestingly, although total proCCK products were normal in the brain they were elevated 3-fold in the intestine, indicating that biosynthesis is upregulated in endocrine cells but not neurons to compensate for the processing defect. These results demonstrate that the CPE mutation differentially affects CCK processing in these two cell types. Intestinal CCK synthesis more closely resembles progastrin processing, suggesting the presence of an endocrine-specific biosynthetic regulatory mechanism not present in neurons.

Animals↗
PubMed1998

Unique progastrin processing in equine G-cells suggests marginal tyrosyl sulfotransferase activity.

Previous studies have indicated that equine G-cell processing of progastrin differs from that of other species. Since the difference may be due to structural features, we have identified equine gastrin-17 and -34 (<ELGLQGSPHLVADLSKKQGPWLEKEEAAYGWMDF-NH2), and cloned a corresponding 451-bp cDNA that encodes a 107-amino-acid preprogastrin. Comparison with other mammalian gastrins shows a high degree of conservation, but instead of four or five acidic residues preceding the bioactive carboxyamidated C-terminal heptapeptide, equine gastrin contains the remarkable substitution of Lys for Glu in this presumed invariant region. In contrast with known mammalian gastrins, which are all significantly Tyr-sulphated, the equine antral gastrins are virtually non-sulphated. Transfection of the equine preprogastrin cDNA into an endocrine cell line resulted in highly sulphated gastrins, indicating that the absence of in situ sulphation is not due to the structure of gastrin, but occurs rather because the equine antral G-cells are unique with respect to tyrosyl sulfotransferase activity. Furthermore, the marginal sulphation may explain the high proportion of gastrin-34 versus gastrin-17 in the equine antrum, since tyrosyl sulphation has been shown to promote the endoproteolytic processing of prohormones.

Amino Acid Sequence↗
PubMed1998

The effect of intermittent injections of CCK-8S and the CCK-A receptor antagonist devazepide on cell proliferation in exocrine rat pancreas.

CONCLUSION: Intermittent injections of sulfated cholecystokinin-8 (CCK-8S) or devazepide caused long-lasting effects on cell proliferation in exocrine pancreas in contrast to continuous infusion. The acinar cells responded to both treatments with changes in the labeling index (LI) during the whole study period. When studying the influence of stimulation and inhibition of the CCK-A receptor on cell proliferation in the exocrine pancreas, not only are the drugs and doses of importance but also the mode of administration. BACKGROUND: Continuous infusion of CCK-8S or the CCK-A receptor antagonist devazepide induces transient changes in acinar cell proliferation in rat pancreas. The aim of the present experiments was to study whether intermittent administration of CCK-8S or devazepide prevents receptor desensitization and thereby affects exocrine pancreatic cell proliferation persistently. METHODS: Male Sprague-Dawley rats were injected subcutaneously twice daily with CCK-8S (6 micrograms), devazepide (240 micrograms) or bovine serum albumin (BSA). The rats were sacrificed after 18 and 36 h and 3 and 7 d. One hour before sacrifice, the rats were injected intraperitoneally with 1 mCi/kg of tritiated thymidine. The pancreatic weight and the contents of water, protein, and DNA were determined. The LI (number of labeled cells/100 cells) of exocrine pancreatic cells was determined microscopically after autoradiography. RESULTS: The concentration of plasma CCK was slightly increased by devazepide, but the increase was more pronounced by CCK-8S. The pancreatic wet weight was transiently increased 18 h after the start of CCK-8S injections (+14%), whereas devazepide caused a reduction after 7 d (-22%). The protein content was uninfluenced and the DNA content was decreased at 36 h with either treatment. CCK-8S increased the LI in acinar and centroacinar cells throughout the study period, but the ductal cell LI was increased only after 18 and 36 h. Injection of devazepide was followed by decreased LI of acinar cells throughout the study period. Also, the centroacinar and ductal cell LI decreased initially but returned to control values after 7 d.

Animals↗
PubMed1998

Processing of precursors of gastroenteropancreatic hormones: diagnostic significance.

The structures and posttranslational maturation patterns of pancreatic and gastrointestinal prohormones are reviewed with emphasis on both general and tumor-specific prohormone processing patterns. Since prohormone processing in neuroendocrine tumors is often attenuated, conventional assays that measure only the phenotypic endpoint of hormone gene expression (i.e., the bioactive hormone) do not quantitate tumor activity accurately. In contrast, measurements that also include prohormones and processing intermediates provide more accurate data on hormone synthesis in gastroenteropancreatic neuroendocrine tumors. To meet such demands we have developed an analytical principle (processing-independent analysis) which quantitates the entire translation product irrespective of the degree of processing. Prospective evaluation in routine diagnostics has shown that processing-independent analysis contributes to ensuring correct diagnosis when gastroenteropancreatic tumors are still small and tumor symptoms uncharacteristic (borderline cases).

Gastrointestinal Hormones↗
PubMed1998

Inhibition of human gastric lipase secretion by glucagon-like peptide-1.

Glucagon-like peptide-1 (GLP-1) may be one of the enterogastrone hormones of the ileal brake mechanism. We therefore studied its effects on gastric lipase secretion in healthy volunteers and vagotomized patients during infusion of pentagastrin. The intestinal incretin hormone GLP-1 (glucagon-like peptide-1, 7-36 amide) was investigated because of its inhibitory effects on gastric acid secretion and motility. GLP-1 infused intravenously in amounts corresponding to the postprandial release significantly inhibited pentagastrin-stimulated gastric lipase secretion and lipolytic activity. The inhibitory effect of GLP-1 persisted in vagotomized patients, suggesting that fundic chief cells, from which gastric lipase is released, or neighboring inhibitory cells could be equipped with GLP-1 receptors. Vagotomized patients had significantly higher plasma concentrations of gastrin and secretin. No significant changes of gastrin, secretin, and CCK secretion were seen during GLP-1 infusion in the vagotomized patients, whereas secretin decreased significantly in the healthy volunteers. GLP-1 seems to be a naturally occurring inhibitor of gastric lipase secretion acting via a nonvagal mechanism. Our results indicate that gastric lipase secretion is subject to hormonal stimulatory as well as inhibitory mechanisms.

Aged↗
PubMed1998

Fasting gall bladder volume and lithogenicity in relation to glucose tolerance, total and intra-abdominal fat masses in obese non-diabetic subjects.

OBJECTIVE: To investigate whether total body fat mass or fat distribution and associated metabolic disturbances in glucose and lipid metabolism influence the well known gallstone pathogenetic factors in obese subjects in order to explain why some obese subjects develop gallstones and some do not. DESIGN: Cross sectional study of gallstone pathogenetic factors, body composition, fat distribution, glucose and lipid metabolism. SUBJECTS: 57 healthy overweight subjects (aged 26-64y, body mass index (BMI) 30-45 kg/m2). MEASUREMENTS: Total and intra-abdominal fat masses were measured by dual X-ray absorptiometry and abdominal CT scanning, respectively. The lithogenic index was measured in aspirated bile. The gallbladder volume was determined by ultrasound and the gallbladder ejection fraction% by dynamic cholescintigraphy. Plasma cholecystokinin (CCK) concentrations during a meal were measured with a specific radioimmunoassay. Insulin sensitivity was measured by the Minimal Model and glucose tolerance by an oral glucose tolerance test (OGTT). Serum lipid concentrations were measured by standard methods. RESULTS: The gallbladder volume in the fasting state increased with increasing intra-abdominal fat mass (P=0.006) and was increased in subjects with impaired glucose tolerance (41 vs 27 ml, P=0.001). The lithogenic index was > 1 in all subjects and correlated with total fat mass (P=0.04). CONCLUSION: Gallstone pathogenesis in obesity seems to be influenced by the total body fat mass and its regional distribution possibly via mutual association with the glucose tolerance.

Abdomen↗
PubMed1998

Regulation of porcine biliary secretion by secretin.

The importance of physiological plasma levels of secretin in biliary bicarbonate secretion is not known. However, in anaesthetized pigs the substantial hepatic output of bicarbonate into the duodenum in response to low doses of secretin exceeds pancreatic bicarbonate output. The aim was therefore to study the relationship between duodenal acidification, secretin and hepatic biliary bicarbonate output in the conscious pig. Göttingen minipigs (n = 22) were cholecystectomized and the common bile duct catheterized. The biliary bicarbonate secretion in response to intraduodenal HCl, secretin or pentagastrin given intravenously, and to meal was studied. Intraduodenal HCl infusion, secretin and pentagastrin given intravenously augmented hepatic bicarbonate output and plasma secretin concentrations significantly. The secretin response to acidification was sufficient to explain the subsequent increase in biliary bicarbonate secretion. Hepatic bicarbonate secretion and concentrations of CCK and secretin in plasma increased postprandially. Exclusion of bile salts from the duodenum abolished postprandial increase in bile volume and increased release of CCK in fasting and fed pigs whereas secretin release was diminished. The results demonstrate that hepatic bicarbonate secretion is stimulated by endogenous secretin and therefore may have a physiological role in duodenal neutralization.

Animals↗
PubMed1998

Renal tubular transport and metabolism of carboxyamidated and glycine-extended gastrins in pigs.

Renal handling of postprandial and intravenously administered gastrin was investigated in anaesthetised pigs. The fractional extraction of postprandial carboxyamidated and glycine-extended gastrin in the kidneys was 0.21 +/- 0.01 and 0.16 +/- 0.02, but the respective urinary clearance comprised only 0.57 +/- 0.03 and 0.44 +/- 0.05% of the GFR (P < 0.02). The respective total body clearance of carboxyamidated and glycine-extended gastrin-17 (gastrin-17 and gastrin-17Gly) during continuous infusion was 22.9 +/- 1.5 and 19.6 +/- 1.4 mL kg-1 min-1 (NS), and the renal fractional extraction of the peptides was 0.31 +/- 0.03 and 0.29 +/- 0.05, respectively. The kidneys accounted for 8% of total body clearance of gastrin-17. Renal filtration rate of gastrin-17 exceeded renal extraction rate (9.739 +/- 0.487 vs. 6.407 +/- 0.321 pmol min-1). Urinary clearance of gastrin-17 and gastrin-17Gly amounted only 0.91 +/- 0.16 and 0.13 +/- 0.03%, respectively, of the GFR (P < 0.01), but urinary excretion rate correlated with the filtered amount of the peptides (r = 0.93, P < 0.01). Neither was a renal plasma threshold recorded nor was a Tm value for tubular uptake or degradation of gastrin achieved in spite of supraphysiological plasma levels of the peptides. The results indicate that filtered gastrin is almost completely removed in the renal tubules, primary by metabolism although part of the absorbed peptides may be returned to the circulation in intact form. The process for uptake or metabolism has a high capacity but varies with the molecular form of gastrin.

Animals↗
PubMed1998

Processing-independent analysis in the diagnosis of gastrinomas.

BACKGROUND: This study evaluates whether a new analytic principle, processing-independent analysis (PIA), offers better specificity and sensitivity than the conventional gastrin radioimmunoassay in the diagnosis of gastrinomas. METHODS: Plasma concentrations of alpha-amidated gastrins and the total progastrin product were measured with radioimmunoassay and with PIA, respectively, in 512 samples taken for gastrin measurement and in a selected group of gastrinoma patients (n=10). RESULTS: Among the 512 patients were 9 with gastrinomas. In plasma from these patients the median degree of amidation (ratio of alpha-amidated gastrins to total progastrin product) was 75% (range, 25-98%), whereas in the other groups the medians varied from 41% to 86%. In the second group of gastrinoma patients all had a degree of amidation of less than 50%. CONCLUSIONS: In screening for gastrinomas PIA offered no diagnostic advantages in comparison with conventional gastrin radioimmunoassay. However, in selected patients who in spite of normal or slightly increased concentrations of amidated gastrins were still suspected of having gastrinoma, additional measurement of the total progastrin product showed incomplete processing of progastrin and thus proved helpful in establishing the diagnosis.

Anti-Ulcer Agents↗
PubMed1998

Impaired gastric acid secretion in gastrin-deficient mice.

To further understand the role of the peptide hormone gastrin in the development and function of the stomach, we have generated gastrin-deficient mice by gene targeting in embryonic stem cells. Mutant mice were viable and fertile, without obvious visible abnormalities. However, gastric function was severely affected by the loss of gastrin. Basal gastric acid secretion was abolished and could not be induced by histamine, carbachol, or gastrin. Histological analysis revealed alterations in the two cell types primarily involved in acid secretion, parietal and enterochromaffin-like (ECL) cells. Parietal cells were reduced in number with an accumulation of immature cells lacking H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase). ECL cells were positioned closer to the base of the gastric glands, with markedly lower expression of histidine decarboxylase. Gastrin administration for 6 days reversed the effects of the gastrin deficiency, leading to an increase in the number of mature, H(+)-K(+)-ATPase-positive parietal cells and a partial restoration of acid secretion. The results show that gastrin is critically important for the function of the acid secretory system.

Animals↗
PubMed1998

The new biology of gastrointestinal hormones.

The classic concept of gastrointestinal endocrinology is that of a few peptides released to the circulation from endocrine cells, which are interspersed among other mucosal cells in the upper gastrointestinal tract. Today more than 30 peptide hormone genes are known to be expressed throughout the digestive tract, which makes the gut the largest endocrine organ in the body. Moreover, development in cell and molecular biology now makes it feasible to describe a new biology for gastrointestinal hormones based on five characteristics. 1) The structural homology groups the hormones into families, each of which is assumed to originate from a common ancestral gene. 2) The individual hormone gene is often expressed in multiple bioactive peptides due to tandem genes encoding different hormonal peptides, alternative splicing of the primary transcript, or differentiated processing of the primary translation product. By these mechanisms, more than 100 different hormonally active peptides are produced in the gastrointestinal tract. 3) In addition, gut hormone genes are widely expressed, also outside the gut. Some are expressed only in neuroendocrine cells, whereas others are expressed in a multitude of different cells, including cancer cells. 4) The different cell types often express different products of the same gene, "cell-specific expression." 5) Finally, gastrointestinal hormone-producing cells release the peptides in different ways, so the same peptide may act as an acute blood-borne hormone, as a local growth factor, as a neurotransmitter, and as a fertility factor. The new biology suggests that gastrointestinal hormones should be conceived as intercellular messengers of general physiological impact rather than as local regulators of the upper digestive tract.

Alternative Splicing↗
PubMed1998

An evaluation of the influence of devazepide and CCK-8S on the intact and resected rat liver.

BACKGROUND: Recently, it has been shown that infusion of the CCK-A receptor antagonist devazepide induced proliferation of hepatocytes and bile duct epithelium in the rat liver. The aim of this study was to further evaluate the influence of devazepide and sulfated cholecystokinin-8 (CCK-8S) on the intact rat liver and rat liver after resection. METHODS: In the first experiment, either saline or devazepide was injected subcutaneously twice daily to rats for 18 and 36 h and 3 and 7 days. In the second experiment, a 70% liver resection was followed by infusion of either DMSO, devazepide, saline or CCK-8S for 2 or 7 days. Prior to sacrifice, all rats received 1 mCi/kg of tritiated thymidine intraperitoneally. The liver was excised and the contents of protein, DNA and water and incorporation of tritiated thymidine were measured. RESULTS: Intermittent injections of devazepide increased the liver protein content after 36 h, followed by a decrease after 7 days. The weight, DNA content or cell proliferation was not affected. Two days after liver resection hyperCCKemia was evoked, which was less prominent after 7 days. Devazepide lowered the plasma concentration of CCK, while the infusion of CCK-8S resulted in extremely high concentrations at both time points. The DNA synthesis measured by thymidine incorporation was increased by devazepide on day 2, whereas the weight or protein and DNA contents of the liver were not influenced. CCK-8S infusion decreased the body and liver weight throughout the study, and the protein and DNA contents after 7 days. CONCLUSIONS: Intermittent devazepide treatment did not affect the intact liver. Devazepide increased the DNA synthesis 2 days after liver resection but was without other influences on the liver regeneration. CCK-8S induced decreased body weight with ensuing negative effects on the liver regeneration. Neither devazepide nor CCK seem to be of any therapeutic use after liver resection or liver failure.

Animals↗
PubMed1998

Accurate measurement of cholecystokinin in plasma.

Shortage of reliable plasma assays has hampered studies of cholecystokinin (CCK). The assay problems are low plasma concentrations, extensive molecular heterogeneity, and close homology of CCK to gastrin, which circulates in higher concentrations. To develop an accurate CCK RIA, antibodies were raised in rabbits, guinea pigs, and mice in titers from 200 to 4000000. The specificity of the antisera was tested with homologous peptides, and tissue and plasma extracts. Rabbit 92128 produced antibodies in high titer (> or =500000) with sufficient avidity (K(o)eff > or = 10(12) mol(-1)) and the desired specificity. The antiserum binds the bioactive forms of CCK with equimolar potency and displays no reactivity with gastrin. CCK concentrations in plasma from healthy humans rose from 1.13 +/- 0.10 pmol/L (mean +/- SE, n = 26) to 4.92 +/- 0.34 pmol/L after a mixed meal. Chromatography of human plasma revealed traces of CCK-58, a predominance of CCK-33 and CCK-22, and moderate amounts of CCK-8. The results show that it is possible to produce specific CCK-antisera using a sulfated CCK-12 analog.

Adult↗