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E S Husebye

Publications and source records attributed to E S Husebye.

31 records · Page 2Linked to original sources
PubMed1997

Chromogranin A and chromogranin B are sensitive circulating markers for phaeochromocytoma.

Specific assays for measurements of circulating chromogranin (Cg) A, CgB, CgC and pancreastatin (Ps) have recently been developed. The aim of the present study was to investigate the usefulness of these markers in diagnosing and following the effects of treatment of patients with phaeochromocytoma, and to compare the results with those concerning other biochemical markers. CgA was elevated in 19/21 (90%), CgB in 17/21 (81%), Ps in 9/21 (43%) and neuropeptide Y in 9/21 (43%) of the patients. Urinary noradrenaline was increased in 19/21 (90%) and urinary adrenaline in 17/19 (89%) of the patients. All patients had increased levels of either urinary catecholamines or plasma chromogranins. In one patient levels of CgA, CgB and Ps were measured at frequent intervals before, during and after surgery. The CgA level fell to normal shortly after the tumour was removed, whereas the CgB level decreased towards normal over the course of several days. Significant correlation was observed between the contents of CgA and CgB in the tumour tissue and the plasma levels of CgA and CgB respectively. We conclude that CgA and CgB are sensitive circulating markers for phaeochromocytoma and that measurements of both urinary catecholamines and plasma chromogranins improve the diagnostic sensitivity. Furthermore, measurements of CgA may be useful in assessing the radicality of surgery in the early postoperative period.

Adolescent↗
PubMed1996

Adrenal autoantibodies and organ-specific autoimmunity in patients with Addison's disease.

OBJECTIVE: Autoimmune destruction of the adrenal gland is the major cause of Idiopathic Addison's disease, but the significance of 21-hydroxylase autoantibodies and their correlation with the presence of other autoantibodies have not so far been investigated in a larger population of patients with Addison's disease. We have now characterized a cohort of patients with idiopathic Addison's disease (n = 97) regarding the specificity of autoantibodies against the adrenal cortex and, as Addison's disease can be either an isolated condition or part of a polyendocrine disorder, we investigated the presence of organ-specific polyendocrine autoimmunity in this patient population. DESIGN: Cross-sectional study. MEASUREMENTS: Autoantibodies were analysed with indirect immunofluorescence (IF) on tissue preparations, ELISA and in Western blots using bacterially expressed proteins. RESULTS: Eighty-four per cent (81/97) of the patient sera recognized the steroid-producing cells of the adrenal cortex in indirect IF. The antigen was identified as 21-hydroxylase by 72% (70/97) of the patient sera in Western blots. Seven sera that were negative on adrenocortical IF identified 21-hydroxylase on Western blot, while eight IF-positive sera were 21-hydroxylase-negative. Five sera weakly recognized 17 alpha-hydroxylase in Western blots, but all of these were also positive for 21-hydroxylase. In 13 cases (12 women), the sera also reacted with testicular Leydig cells, and nine of these identified the side-chain cleavage (SCC) enzyme. Other clinically evident organ-specific autoimmune disorders were present in 40% of the 97 patients and abnormal titres of organ-specific antibodies were found in 60% of the patients. CONCLUSIONS: In idiopathic Addison's disease, auto-antibodies against 21-hydroxylase are found in a majority of cases and this represents an important diagnostic tool. The enzyme 17 alpha-hydroxylase does not seem to constitute a major autoantigen in Addison's disease. In a subgroup of patients with autoantibodies to gonads, antibodies to SCC are produced, often in parallel with antibodies to 21-hydroxylase. In yet another subgroup the specificity of autoantibodies giving positive immunofluorescence is still unknown. Three patients revealed a polyendocrine syndrome which clinically resembles autoimmune polyendocrine syndrome (APS) type I, but serologically corresponds to APS type II. Polyendocrine disorders are often associated with Addison's disease, and screening, including quantification of autoantibodies, may help to identify those at risk of developing associated autoimmune disorders.

Addison Disease↗
PubMed1996

Autoantibodies against adrenal medulla in type 1 and type 2 diabetes mellitus: no evidence for an association with autonomic neuropathy.

OBJECTIVE: To examine the role of autoimmunity in the development of autonomic neuropathy in diabetes mellitus. DESIGN: Retrospective cross-sectional study. SETTING: The Department of Medicine, University Hospital, Uppsala, and the Department of Endocrinology, University of Lund, Malmö General Hospital, Malmö, Sweden. SUBJECTS AND MAIN OUTCOME MEASURES: We examined 135 patients with type 1 (n = 96) or type 2 (n = 39) diabetes mellitus. Tests for cardiovascular autonomic functions were performed, and patient sera were analysed for the presence of autoantibodies against the adrenal medulla by indirect immunofluorescence, Western blot and immunoprecipitation techniques. RESULTS: Sera from 13% (12/96) of the type 1 and 20% (8/39) of the type 2 patients showed a positive cytoplasmic immunofluorescence (IF) staining of the adrenal medullary cells, as did 20% (30/151) of sera from healthy controls. Fifty-eight and 64% of type 1 and type 2 patients, respectively, had cardiovascular autonomic neuropathy, but no correlation between autonomic neuropathy and positive IF against the adrenal medulla was observed, with the exception of significant drops in diastolic blood pressure 8 min after tilt (P < 0.005) in type 1 patients. The various IF-positive sera reacted with several different proteins when analysed with Western blot technique using a total homogenate of the bovine adrenal medulla. These did not correspond to any of the proteins involved in the synthesis or storage of catecholamines which were considered as putative autoantigens. CONCLUSION: The finding of similar frequencies of immunoglobulins binding to adrenal medulla in both type 1 and 2 diabetic patients as well as in normal controls, argues against a role of anti-adreno-medullary antibodies in the pathogenesis of autonomic diabetic neuropathy.

Adrenal Medulla↗
PubMed1995

Aromatic-L-amino-acid decarboxylase, a pyridoxal phosphate-dependent enzyme, is a beta-cell autoantigen.

Different autoantigens are thought to be involved in the pathogenesis of insulin-dependent diabetes mellitus, and they may account for the variation in the clinical presentation of the disease. Sera from patients with autoimmune polyendocrine syndrome type I contain autoantibodies against the beta-cell proteins glutamate decarboxylase and an unrelated 51-kDa antigen. By screening of an expression library derived from rat insulinoma cells, we have identified the 51-kDa protein as aromatic-L-amino-acid decarboxylase (EC 4.1.1.28). In addition to the previously published full-length cDNA, forms coding for a truncated and an alternatively spliced version were identified. Aromatic L-amino acid decarboxylase catalyzes the decarboxylation of L-5-hydroxytryptophan to serotonin and that of L-3,4-dihydroxyphenylalanine to dopamine. Interestingly, pyridoxal phosphate is the cofactor of both aromatic L-amino acid decarboxylase and glutamate decarboxylase. The biological significance of the neurotransmitters produced by the two enzymes in the beta cells remains largely unknown.

Adolescent↗
PubMed1990

The effect of prenylamine and organic nitrates on the bioenergetics of bovine catecholamine storage vesicles.

We have compared the cardioprotective agents prenylamine and glyceryl trinitrate (GTN) with respect to their effects on the bioenergetics of catecholamine storage vesicles. Chromaffin granule ghosts, which have a well preserved ability to actively transport and store catecholamines, were used as a model for adrenergic synaptic vesicles due to their functional similarity. Prenylamine, which partially and reversibly deplete the endogenous stores of noradrenaline in adrenergic nerves and ganglia, was found to inhibit the generation of the transmembrane proton electrochemical gradient driven by a H(+)-ATPase, mainly by acting as an uncoupler of this ATPase. The inhibition of the energy dependent dopamine uptake (and noradrenaline biosynthesis) by prenylamine could be accounted for by its effect on the bioenergetics of the storage vesicles. The organic nitrates glyceryl trinitrate and isosorbide dinitrate also partly inhibited the catecholamine uptake in parallel with their effects on the proton electrochemical gradient. It is concluded that GTN is a weak catecholamine depletor. Experiments with 3-morpholinosydnonimin-hydrochloride, a source of nitric oxide (NO), opens up the possibility that the mechanism of inhibition of the bioenergetics of chromaffin granule ghosts by GTN is mediated by NO.

Animals↗
PubMed1990

Purification and kinetic properties of a membrane-bound phosphatidylinositol kinase of the bovine adrenal medulla.

Phosphatidylinositol (PI) kinase (EC 2.7.1.67), an integral membrane protein of chromaffin granule ghosts of the bovine adrenal medulla, was found to phosphorylate PI in the 4-position of the inositol ring. The PI kinase was purified about 200-fold from a membrane fraction containing chromaffin granules and microsomes by extraction with Triton X-114, followed by phase partition (clouding) and heparin Sepharose chromatography. The PI kinase preparation (specific activity of 5.1 nmol PIP/mg protein per min) was free from other enzymatic activities that metabolize polyphosphoinositides. Km values of 55 microM and 40 microM for ATP and PI, respectively, were estimated for the purified enzyme. Concentrations of Triton X-100 above the critical micellar concentration (0.01%, w/v) were necessary to support significant enzyme activity, which was optimal at about 0.1% (w/v). Its dependence of pH was similar to that of the membrane-bound enzyme, with a broad optimum around pH 7. Mes in the millimolar concentration range was found to strongly inhibit the activity of the purified PI kinase (I50 at about 4 mM). The enzyme was almost totally inhibited by low micromolar concentrations of free calcium, and stimulated by hydrophilic cations, e.g., Mg2+ and poly(L-lysine), with the same potencies as for the membrane-bound enzyme. The amphiphilic cation trifluoperazine, however, stimulated the activity of purified PI kinase less effectively than the membrane-bound enzyme (Husebye, E.S. and Flatmark, T. (1988) Biochem. Pharmacol. 37, 449-456), whereas the inhibitory effect of near millimolar concentrations of trifluoperazine was the same for the two forms of the enzyme. It is concluded that the membrane-bound PI kinase of this tissue is of type II according to the classification of Cantley and co-workers (Whitman et al. (1987) Biochem. J. 247, 165-174).

1-Phosphatidylinositol 4-Kinase↗
PubMed1989

Purification and kinetic properties of a soluble phosphatidylinositol-4-phosphate kinase of the bovine adrenal medulla with emphasis on its inhibition by calcium ions.

A phosphatidylinositol-4-phosphate kinase (PIP kinase, EC 2.7.1.68) was purified about 1000-fold from the cytosolic fraction of bovine adrenal medulla by ammonium sulfate precipitation, anion exchange, phosphocellulose and gel permeation chromatography. The PIP kinase preparation was free from other polyphosphoinositide metabolizing activities, and some of its catalytic properties were studied using both membrane-bound and liposomal/micellar phosphatidylinositol 4-phosphate (PIP) as the substrate. Using a native substrate (chromaffin granule ghosts) enriched in PIP, the Km value for MgATP was about 130 microM at 5 mM Mg2+, and optimum activity was obtained at pH 7.2. ADP inhibited the PIP kinase activity in a mixed non-competitive manner. The enzyme was stimulated by Mg2+ and spermidine, whereas free calcium effectively inhibited the PIP kinase activity (I50 at about 0.1 microM Ca2+). This inhibition was independent of calmodulin and CaATP (80 microM) was not a substrate for the PIP kinase. The presence of near intracellular concentrations of potassium decreased the sensitivity to calcium ions (I50 of about 50 microM), but the inhibitory effect was still evident in the physiologically interesting concentration range. The concentration for half-maximal inhibition was three orders of magnitude higher (I50 of about 200 microM) using a liposomal/micellar suspension of purified PIP as the substrate, which demonstrates the importance of using a native substrate in studying the regulatory properties of this enzyme. It is concluded that the high sensitivity of the PIP kinase activity to calcium ions is likely to be physiologically significant, as recently proposed for the phosphatidylinositol (PI) kinase (Husebye, E.S. and Flatmark, T. (1988) Biochim. Biophys. Acta 968, 261-265), and may represent a negative feedback control of the cytosolic calcium concentration. The cationic amphiphile trifluoperazine was a potent inhibitor of PIP kinase activity (I50 at about 15 microM), and may represent a useful tool to study PI kinase selectively in membranes or cells containing both kinases.

Adrenal Medulla↗
PubMed1988

Phosphatidylinositol kinase of bovine adrenal chromaffin granules. Modulation by hydrophilic and amphiphilic cations.

The effects of hydrophilic and amphiphilic cations on the activity of phosphatidylinositol (PI) kinase (EC 2.7.1.67) of chromaffin granule ghosts were investigated. The cations studied can be divided into two groups, i.e. (i) compounds with a biphasic response (stimulation and inhibition), and (ii) those with a selective stimulatory effect on the enzyme activity. The cationic amphiphile trifluoperazine belongs to the first group, and stimulated the enzyme activity, maximal at 80 microM (2-fold), with a progressive inhibition at higher concentrations. This biphasic response was shared by a number of structurally related cationic amphiphiles, i.e. the tricyclic antidepressants, imipramine and desipramine, the phenothiazine, chlorpromazine, the miconazole derivative, calmidazolium, the beta-adrenergic agonist, propranolol, compound 48/80, as well as by the hydrophilic cations neomycin and poly-L-lysine. On the other hand, a pure stimulatory effect was observed with the amphiphilic polypeptide mastoparan and the polycationic compound spermidine, whereas ACTH1-39 and ACTH1-24 (peptides structurally related to mastoparan) revealed a slight inhibitory effect. We conclude that all the cations tested including Mg2+, stimulate PI kinase activity rather unspecifically by binding of the positively charged groups to a membrane component, probably the PI kinase itself. This site is different from that mediating the specific inhibition by calcium (Husebye ES and Flatmark T, Biochim Biophys Acta 968: 261-265, 1988). The inhibitory effect of cationic amphiphiles is correlated to their lipid solubility, and represents a perturbation of the membrane structure, but not a solubilization of enzyme or phosphoinositide from the membrane. The inhibitory effect of hydrophilic cations is due to complexation of ATP.

1-Phosphatidylinositol 4-Kinase↗
PubMed1988

Phosphatidylinositol kinase of bovine adrenal chromaffin granules: kinetic properties and inhibition by low concentrations of Ca2+.

The kinetics and regulatory properties of phosphatidylinositol (PI) kinase were studied in chromaffin granule ghosts isolated from the bovine adrenal medulla. Phosphatidylinositol 4-phosphate (PIP) was the major 32P-labelled phospholipid formed when the isolated membranes were phosphorylated by [gamma-32P]ATP. The PI kinase activity was rather independent of pH, but highly dependent on Mg2+ with a maximal stimulation at 60 mM Mg2+. By contrast, KCl and NaCl had a slight inhibitory effect. The Km value for MgATP was 44 and 62 microM in the presence of 1 and 20 mM MgCl2, respectively. The PI kinase was almost fully and reversibly inhibited by free Ca2+ (calmodulin-independent) in the nanomolar and low micromolar range, depending on the concentration of Mg2+. The inhibition was not dependent on Ca2+-stimulated protein phosphorylation, and it could not be explained by a dephosphorylation of PIP.

1-Phosphatidylinositol 4-Kinase↗
PubMed1987

Characterization of phospholipase activities in chromaffin granule ghosts isolated from the bovine adrenal medulla.

Highly purified chromaffin granule membranes contain high levels (100 nmol/mg protein) of long-chain free fatty acids (Husebye, E.S. and Flatmark, T. (1984) J. Biol. Chem. 259, 15272-15276), as well as lysophosphatidylcholine (268 nmol/mg protein) and lysophosphatidylethanolamine (92 nmol/mg protein). The release of saturated and unsaturated long-chain fatty acids from endogenous phospholipids was 38 and 28 nmol/mg protein per h, respectively, at 37 degrees C and pH 7.5 (alkaline pH optimum). p-Bromophenacyl bromide inhibited the release of palmitate and oleate by 88 and 65%, respectively. The deacylation of membrane phospholipids was not significantly affected by micromolar free Ca2+. Based on experiments with pancreatic phospholipase A2, stearate and arachidonate were found to be suitable markers for deacylation at the sn-1 and sn-2 positions, respectively. Experiments with exogenously added labeled phosphatidylcholines confirmed that chromaffin granule ghosts contain a phospholipase A2 activity (alkaline pH optimum). The preparations also revealed a phospholipase A1 activity (acid pH optimum). Finally, the ghosts contain a lysophospholipase activity (alkaline pH optimum), that accounts for the major part of the deacylation of membrane phospholipids, notably the release of saturated fatty acids (stearate and palmitate). It is unlikely that the high content of lysophospholipids is an artifact of the procedure by which the granule ghosts are isolated.

Acylation↗
PubMed1984

The content of long-chain free fatty acids and their effect on energy transduction in chromaffin granule ghosts.

Highly purified chromaffin granule ghosts contain about 100 (89.1-110.2, n = 8) nmol of long-chain free fatty acids/mg of protein. About 40% of the fatty acids were mono- and polyunsaturated, of which arachidonate accounted for approximately 20%. Incubation with fatty acid-free bovine serum albumin selectively depleted the ghosts of unsaturated fatty acids. The effect of a series of long-chain fatty acids on energy coupling was studied in resealed chromaffin granule ghosts energized by MgATP. Their effectiveness in inhibiting the generation of the transmembrane proton concentration gradient and transmembrane potential was found to be correlated with their chain length and degree of unsaturation, all-cis-5,8,11,14-icosatetraenoate (arachidonate) and all-cis-5,8,11,14,-17-icosapentaenoate being most effective. Arachidonate was also found to dissipate an artificially generated pH gradient. Treatment of chromaffin granule ghosts with bovine serum albumin increased the transmembrane proton concentration gradient that could be generated by MgATP. This enhanced response to MgATP is presumably due to the removal of free fatty acids from the ghosts. Considering the potency of arachidonate and other unsaturated fatty acids, as well as their high endogenous content, these "uncoupling agents" should be accounted for when isolated chromaffin granules are used for in vitro studies on energy coupling.

Adenosine Triphosphatases↗