Reference gene selection in human periodontal ligament cells

Background: Real-time quantitative PCR (qPCR) is a highly sensitive and specific method which is used extensively for determining gene expression profiles in a variety of cell and tissue types. In order to obtain accurate and reliable gene expression quantification, qPCR data are generally normalised against so-called reference or housekeeping genes. Ideally, reference genes should have abundant and stable RNA transcriptomes under the experimental conditions employed. However, reference genes are often selected rather arbitrarily and indeed some have been shown to have variable expression in a variety of in vitro experimental conditions.
Objective: The objective of the current study was to investigate reference gene expression in human periodontal ligament (PDL) cells in response to treatment with lipopolysaccharide (LPS).
Method: Primary human PDL cells were grown in Dulbecco’s Modified Eagle Medium with L-glutamine supplemented with 10% fetal bovine serum, 100UI/ml penicillin and 100µg/ml streptomycin. RNA was isolated using the RNeasy Mini Kit (Qiagen) and reverse transcribed using the QuantiTect Reverse Transcription Kit (Qiagen). The expression of a total of 19 reference genes was studied in the presence and absence of LPS treatment using the Roche Reference Gene Panel. Data were analysed using NormFinder and Bestkeeper validation programs.
Results: Treatment of human PDL cells with LPS resulted in changes in expression of several commonly used reference genes, including GAPDH. On the other hand the reference genes β-actin, G6PDH and 18S were identified as stable genes following LPS treatment.
Conclusion: Many of the reference genes studied were robust to LPS treatment (up to 100 ng/ml). However several commonly employed reference genes, including GAPDH varied with LPS treatment, suggesting they would not be ideal candidates for normalisation in qPCR gene expression studies.

Gendered Candidate Selection and the Representation of Women in Northern Ireland

In an attempt to account for the exceptionally low levels of female representation in Northern Ireland, this paper provides an analysis of the contemporary candidate selection procedures of the region's five main political parties. Drawing on evidence gathered from 29 elite interviews, plus official internal party documents, the study finds that the localised nature of the parties' selection procedures may disadvantage women aspirants. Also important are ‘supply-side’ factors influencing legislative recruitment and female participation rates, namely the strongly embedded social norm of female domestic responsibility, a masculinised political culture and the lack of confidence of potential female candidates.

The Selection of Party Leaders in Northern Ireland

This article provides an analysis of the leadership selection methods adopted by Northern Ireland's five main parties. Drawing on data from interviews with party elites and internal party documents, it sheds light on an important element of intra-party organisation in the region and constitutes a rare case-study of leadership selection in a consociational democracy. By accounting for instances of organisational reform, this article also reveals the extent to which Northern Ireland's parties align with the wider comparative trend of leadership ‘democratisation’. In terms of ‘who’ selects party leaders, the analysis finds a substantial degree of organisational heterogeneity and a reasonably high rate of democratisation. Northern Ireland's parties also prove rather exceptional in their universal adoption of short fixed terms for party leaders and, in the case of three of the parties, their preference for high candidacy thresholds.

Selection of the most appropriate roadside vehicle restraint system – the SAVeRS project

Run Off Road (ROR) crashes are road accidents that often result in severe injuries or fatalities. To reduce the severity of ROR crashes, “forgiving roadsides” need to be designed and this includes identifying situations where there is a need for a Vehicle Restraint System (VRS) and what appropriate VRS should be selected for a specific location and traffic condition. Whilst there are standards covering testing, evaluation and classification of VRS within Europe (EN1317 parts 1 to 8), their selection, location and installation requirements are typically based upon national guidelines and standards, often produced by National Road Authorities (NRA) and/or overseeing organisations. Due to local conditions, these national guidelines vary across Europe.
The European SAVeRS project funded by CEDR has developed a practical and readily understandable VRS guidance document and a user-friendly software tool which allow designers and road administrations to select the most appropriate solution in different road and traffic conditions.
This paper describes the main outcomes of the project, the process to select the most appropriate roadside barrier, and the user friendly SAVeRS tool.