Quantitation of alternatively spliced NMDA receptor NR1 isoform mRNA transcripts in human brain by competitive RT-PCR

The N-methyl-D-aspartate (NMDA)-selective subtype of ionotropic glutamate receptor is of importance in neuronal differentiation and synapse consolidation, activity-dependent forms of synaptic plasticity, and excitatory amino acid-mediated neuronal toxicity [Neurosci. Res. Program, Bull. 19 (1981) 1; Lab. Invest. 68 (1993) 372]. NMDA receptors exist in vivo as tetrameric or pentameric complexes comprising proteins from two families of homologous subunits, designated NR1 and NR2(A-D) [Biochem. Biophys. Res. Commun. 185 (1992) 826]. The gene coding for the human NR1 subunit (hNR1) is composed of 21 exons, three of which (4, 20 and 21) can be differentially spliced to generate a total of eight distinct subunit variants. We detail here a competitive RT-PCR (cRT-PCR) protocol to quantify endogenous levels of hNR1 splice variants in autopsied human brain. Quantitation of each hNR1 splice variant is performed using standard curve methodology in which a known amount of synthetic ribonucleic acid competitor (internal standard) is co-amplified against total RNA. This method can be used for the quantitation of hNR1 mRNA levels in response to acute or chronic disease states, in particular in the glutamatergic-associated neuronal loss observed in Alzheimer's disease [J. Neurochem. 78 (2001) 175]. Furthermore, alterations in hNR1 mRNA expression may be reflected at the translational level, resulting in functional changes in the NMDA receptor. (C) 2003 Elsevier Science B.V. All rights reserved.

Quantitation of human brain GABAA receptor isoforms by competitive RT-PCR

We have developed a competitive RT-PCR assay, adapted from Lewohl et al. [Brain Res. Brain Res. Protoc. 1 (1997) 347]. for the quantitation of GABA, receptor beta isoforms in human brain using an internal standard that shares high sequence homology to the targets. The internal standard is identical to the beta(1) sequence except for a 61 bp deletion and the incorporation of a Hind III restriction enzyme site. Unlike traditional competitive RT-PCR, which requires a range of internal standard concentrations to be titrated against a constant amount of unknown, this method relies on a standard curve for quantitation of each sample and thus permits increased sample throughput. This method is suitable for the quantitation of beta(1), beta(2) and beta(3) isoforms of the GABA(A) receptor in human alcoholic and control brain. (C) 2003 Elsevier Science B.V. All rights reserved.

The case for emotional intelligence in organizational research

Since its popularization by Goleman (1995), the concept of emotional intelligence has been the subject of ongoing controversy, so it is understandable that the model we proposed, which includes emotional intelligence as a moderator variable, would attract its share of criticism.

Is emotional intelligence training for leaders justified?

In this paper, we present the results of a qualitative study of subordinate perceptions of leaders. The study represents a preliminary test of a model based on Affective Events Theory, which posits that leaders who are seen to be effective shape the affective events that determine employees' attitudes and behaviours in the workplace. Within this framework, we argue that effective leaders ameliorate employees' hassles by providing frequent, small emotional uplifts. The resulting positive affective states are then proposed to lead to more positive employee attitudes and behaviours, and more positive regard for the leader. Importantly, leaders who demonstrate these ameliorating behaviours are likely to require high levels of emotional intelligence, defined in terms of the ability to recognise, understand, and manage emotions in self and others. To investigate this model, we conducted interviews and focus groups with 10 leaders and 24 employees. Results confirmed that these processes do indeed exist in the workplace. In particular, leaders who were seen by employees to provide continuous small emotional uplifts were consistently held to be the most effective. Study participants were especially affected by negative events (or hassles). Leaders who failed to deal with hassles or, worse still, were the source of hassles, were consistently seen to be less effective. We conclude with a discussion of implications for practicing managers, and suggest that our exploratory findings provide justification for emotional intelligence training as a means to improve leader perceptions and effectiveness. [Abstract from author]