Cent Of Total Radioactivity Research Articles

Modulation of gastrin-releasing peptide (GRP) receptors in insulin secreting cells.

Gastrin-releasing peptide (GRP) receptors are present in pancreatic islets, though their regulation is unknown except for homologous desensitization. The modulation of binding of GRP to mouse pancreatic islets and INS-1 cells was studied. At 60 min (steady-state), total binding of [(125)I-Tyr(15)] GRP was 1.62 per cent of total radioactivity per 50 islets; non-specific binding (presence of 1 mM unlabelled GRP(1-27)) was 0.05 to 0.61 per cent of total radioactivity. A preincubation with 1000 nM cholecystokinin (CCK(8)) or with 1000 nM glucose-dependent insulinotropic peptide (GIP) augmented the number of GRP binding sites but not their affinity. [(125)I-Tyr(15)]GRP binding to INS-1 cells was saturable (90 min) and specific with respect to compounds that are not chemically related to GRP (e.g. calcitonin gene-regulated peptide-CGRP and atrial natriuretic peptide-ANP). Displacement studies showed one binding site with a K(d) of 0.39 nM and a B(max) of 13.2 fmoles mg(-1) protein. When the cells were pretreated for 24 h with 10 nM GIP or CCK(8), only GIP but not CCK(8) increased the B(max) of the GRP binding site. The affinity (K(d)) was not changed by either compound. This effect of GIP pretreatment was not affected by downregulating PKC by TPA (phorbol ester; long-term pretreatment). These data indicate that: (1) specific binding sites for GRP are present in mouse pancreatic islets and INS-1 cells; (2) the GRP binding is upregulated by GIP in both islets and INS-1 cells and additionally by CCK(8 ), albeit only in islets; and (3) PKC does not seem to be involved in the up-regulation process. Thus a positive interplay between both the incretins GIP and CCK(8) and the neurotransmitter GRP is obvious.

The metabolism of iodine-123 labelled 3-(5-cyclopropyl-1,2,4-oxadiazo-3-yl)-7-iodo-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]benzodiazepine (NNC 13-8241) measured in human plasma is only minor

The imidazobenzodiazepine derivate 123I-labelled 3-(5-cyclopropyl-1,2,4-oxadiazo-3-yl)-7-iodo-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a][1,4]-benzodiazepine (NNC 13–8241) is a partial benzodiazepine agonist and binds with a high affinity to the benzodiazepine receptor. The favourable kinetic properties indicate that [123I]NNC 13–8241 is a promising SPET ligand. In the present study, an extensive examination of the metabolite pattern of [123I]NNC 13–8241 in plasma and urine from seven healthy subjects was performed using gradient HPLC. After injection of [123I]NNC 13–8241 into human beings, only one radioactive metabolite was found in plasma 3–300 min post injection. This polar metabolite eluted together with the solvent front fraction. The proportion of unchanged [123I]NNC 13–8241 was 82–86 per cent during the entire study. In addition, two other radioactive metabolites were found in urine. The first metabolite was lipophilic and eluted slightly before the parent compound [123I]NNC 13–8241. The second metabolite eluted slightly after solvent front peak. The amount of unchanged [123I]NNC 13–8241 in human urine was 9 per cent and the solvent front fraction contained 72 per cent of total radioactivity. No detectable radioactivity appeared with the same retention time as synthetisized nor-NNC 13–8241 in plasma or urine, which excludes the possibility of in vivo demethylation of [123I]NNC 13–8241. In conclusion, the [123I]NNC 13–8241 was found to have only a minor metabolism, which favours its use as a SPET tracer for quantitation of the benzodiazepine receptor. © 1998 John Wiley & Sons, Ltd.

Studies on Cassava, Manihot utilissima. II. Biosynthesis of Asparagine‐14C from 14C‐labelled Hydrogen Cyanide and its Relations with Cyanogenesis

AbstractSeeds and seedlings of Manihot utilissima were analysed for cyanogenic glycosides und free amino acids, with special reference to valine and isoleucine which serve as precursors of the aglycone moieties of linamarin and lotaustralin. Seeds contained traces of valine and isoleucine but no glycosides, whereas seedlings contained high concentrations of these amino acids and glycosides. Illumination of seedlings led to a steep increase in the concentration of glycosides followed by a decrease without excretion of detectable HCN. Seeds accumulated asparagine, while seedlings accumulated both asparagine and glutamine in the storage and transport of nitrogen. Seedlings incorporated 13.2 per cent of label from valine‐14C(U) and 2.4 per cent of label from isoleucine‐14C(U)into linamarin and lotaustralin, respectively. In both cases, appreciable amounts of label were also incorporated into asparagine. 49 per cent of label from H14CN was incorporated inio asparagine in which ca. 98 per cent of total radioactivity was located in the amide‐carbon atom. The different patterns of labelling which occurred during the assimilation of H14CN and 14CO2 showed that cyanide metabolism did not proceed via CO2, and that M. utilissima contains an efficient enzyme‐system which catalyses the conversion on high concentrations of HCN into asparagine, which subsequently enters different metabolic pools involved with respiration, protein and carbohydrate syntheses. Cyanogenesis in M. utilissima appears lo be directly influenced by available pools of valine and isoleucine, and the metabolism of HCN released from linamarin and lotaustralin by the action of linamarase may be directly related to respiratory and synthetic processes by way of the incorporation of HCN as a unit into asparagine.