Characterization and effects on cAMP accumulation of adrenomedullin and calcitonin gene-related peptide (CGRP) receptors in dissociated rat spinal cord cell culture

1 Adrenomedullin (AM) and calcitonin gene-related peptide (CGRP) have structural similarities, interact with each others receptors (calcitonin receptor-like receptor (CLR)/receptor-activity-modifying proteins (RAMPs)) and show overlapping biological activities. AM and CGRP receptors are chiefly coupled to cAMP production. In this study, a method of primary dissociated cell culture was used to investigate the presence of AM and CGRP receptors and their effects on cAMP production in embryonic spinal cord cells. 2 Both neuronal and non-neuronal CLR immunopositive cells were present in our model. 3 High affinity, specific [ 125I]-AM binding sites (K(d) 79±9 pM and B(max) 571±34 fmol mg -1 protein) were more abundant than specific [ 125I]-CGRP binding sites (K(d) 12±0.7 pM and B(max) 32±2 fmol mg -1 protein) in embryonic spinal cord cells. 4 Specific [ 125I]-AM binding was competed by related molecules with a ligand selectivity profile of rAM>hAM(22-52)>rCGRPα>CGRP(8-37) ≫[r-(r*,s*)]-N-[2-[[5-amino-1-[[4-(4-pyridinyl)-1-piperazinyl] carbonyl]pentyl]amino]-1-[(3,5-dibromo-4-hydroxyphenyl)methyl]-2-oxoethyl]-4-(1, 4-dihydro-2-oxo-3(2H)-quinazolinyl)-,1-piperidinecarboxamide (BIBN4096BS). 5 Specific [ 125I]-CGRP binding was competed by rCGRPα>rAM≥ CGRP(8-37)≥BIBN4096BS>hAM(22-52). 6 Cellular levels of cAMP were increased by AM (pEC"5"0 10.2±0.2) and less potently by rCGRPα (pEC"5"0 8.9±0.4). rCGRPα-induced cAMP accumulation was effectively inhibited by CGRP(8-37) (pA"2 7.63±0.44) and hAM(22-52) (pA"2 6.18±0.21) while AM-stimulation of cAMP levels was inhibited by CGRP(8-37) (pA"2 7.41±0.15) and AM(22-52) (pA"2 7.26±0.18). BIBN4096BS only antagonized the effects of CGRP (pA"2 8.40±0.30) on cAMP accumulation. 7 These pharmacological profiles suggest that effects of CGRP are mediated by the CGRP"1 (CLR/RAMP1) receptor in our model while those of AM are related to the activation of the AM"1 (CLR/RAMP2) receptor subtype. © 2006 Nature Publishing Group All rights reserved.

Safety culture during major organisational change

This research examines the effect of major changes, in the external context, on the safety culture of a UK generating company. It was focused on an organisation which was originally part of the state owned Central Electricity Generating Board and which, by the end of the research period, was a self-contained generating company, operating in a competitive market and a wholly owned subsidiary of a US utility. The research represents an attempt to identify the nature and culture of the original organisation and to identify, analyse and explain the effects of the forces of change in moulding the final organisation. The research framework employed a qualitative methodology to investigate the effects of change, supported by a safety culture questionnaire, based on factors identified in the third report of the ACSNI Human Factors Study Group; Organising for Safety, as being indicators of safety culture. An additional research objective was to assess the usefulness of the ACSNI factors as indicators of safety culture. Findings were that the original organisation was an engineering dominated technocracy with a technocentric safety culture. Values and beliefs were very strongly held and resistant to change and much of the original safety culture survived unchanged into the new organisation. The effects of very long periods of uncertainty about the future were damaging to management/worker relationships but several factors were identified which effectively insulated the organisation from any of the effects of change. The forces of change had introduced a beneficial appreciation of the crucial relationship between safety risk assessment and commercial risk assessment.Although the technical strength of the original safety culture survived, so did the essential weakness of a low level of appreciation of the human behavioural aspects of safety. This led to a limited, functionalist world view of safety culture, which assumed that cultural change was simpler to achieve than was the case and an inability to make progress in certain areas which were essentially behavioural problems.The factors identified by ACSNI provided a useful basis for the site research methodology and for identifying areas of relative strength and weakness in the site safety arrangements.