A novel diagnostic test detects a low frequency of the hemicentin Gln5345Arg variant among Northern Irish age related macular degeneration patients.

Purpose: Age related macular degeneration (AMD) is a common cause of severe vision loss. Identification of genes involved in AMD will facilitate early detection and ultimately help to identify pathways for treatment for this disorder. The A16,263G mutation in the HEMICENTIN-1 gene produces a non-conservative substitution of arginine for glutamine at codon 5345 which has been implicated in familial AMD. The aim of this study is to develop a rapid diagnostic assay for the detection of this mutation and to evaluate its frequency in a sample of AMD patients. Methods: A primer probe set was designed from exon 104 of the HEMICENTIN-1 gene to differentiate between mutant and wild type alleles. A region spanning the mutation was amplified by PCR using a LightCycler (Roche Diagnostic). The mutation was then detected by melt curve analysis of the hybrid formed between the PCR product and a specific fluorescent probe. The frequency of the mutation within the Northern Ireland population was evaluated by assaying 508 affected AMD patients, 25 possibly affected and 163 controls. Results: This assay clearly discriminates between the A16,263G mutant and wild type HEMICENTIN-1 alleles. The wild type sequence has a single base mismatch with the probe which decreases the stability of the hybrid, resulting in a lower TM (TM=51.27 °C) than that observed for the perfectly matched mutant allele (TM=59.9 °C). The mutant allele was detected in only one of the 696 subjects, an affected AMD patient. Conclusions: We describe a rapid assay for the genotyping of the Gln5345Arg mutation using real-time fluorescence PCR to facilitate rapid processing of samples through combined amplification and detection steps. These characteristics are suitable for a clinical setting where high throughput diagnostic procedures are required. The frequency of this mutation within the Northern Ireland population has been estimated at 0.2%, concurring with previous findings that this mutation is a rare variant associated with AMD. A rapid diagnostic assay will facilitate a reliable and convenient evaluation of the frequency of the Gln5345Arg mutation and its association with AMD within other populations.

Lack of MEF2A Δ7aa mutation in Irish families with early onset ischaemic heart disease, a family based study

Background: Ischaemic heart disease (IHD) is a complex disease due to the combination of environmental and genetic factors. Mutations in the MEF2A gene have recently been reported in patients with IHD. In particular, a 21 base pair deletion (Δ7aa) in the MEF2A gene was identified in a family with an autosomal dominant pattern of inheritance of IHD. We investigated this region of the MEF2A gene using an Irish family-based study, where affected individuals had early-onset IHD. Methods: A total of 1494 individuals from 580 families were included (800 discordant sib-pairs and 64 parent-child trios). The Δ7aa region of the MEF2A gene was investigated based on amplicon size. Results: The Δ7aa mutation was not detected in any individual. Variation in the number of CAG (glutamate) and CCG (proline) residues was detected in a nearby region. However, this was not found to be associated with IHD. Conclusion: The Δ7aa mutation was not detected in any individual within the study population and is unlikely to play a significant role in the development of IHD in Ireland. Using family-based tests of association the number of tri-nucleotide repeats in a nearby region of the MEF2A gene was not associated with IHD in our study group. © 2006 Horan et al; licensee BioMed Central Ltd.

Design of granular pavements

The use of high-quality quarried crushed rock aggregates is generally required to comply with current specifications for unbound granular materials (UGMs) in pavements. The source of these high-quality materials can be a long distance from the site, resulting in high transportation costs. The use of more local sources of marginal materials or the use of secondary aggregates is not allowed if they do not fully comply with existing specifications. These materials can, however, be assessed for their suitability for use in a pavement by considering performance criteria such as resistance to permanent deformation and degradation instead of relying on compliance with inflexible specifications. The final thickness of the asphalt cover and the pavement depth are governed by conventional pavement design methods, which consider the number of vehicle passes, subgrade strength, and some material property, commonly the California bearing ratio or resilient modulus. A pavement design method that includes as a design criterion an assessment of the resistance to deformation of a UGM in a pavement structure at a particular stress state is proposed. The particular stress state at which the aggregate is to perform in an acceptable way is related to the in situ stress, that is, the stress that the aggregate is anticipated to experience at a particular depth in the pavement. Because the stresses are more severe closer to the pavement surface, the aggregates should be better able to resist these stresses the closer they are laid to the surface in the pavement. This method was applied to two Northern Ireland aggregates of different quality (NI Good and NI Poor). The results showed that the NI Poor aggregate performed at an acceptable level with respect to permanent deformation, provided that a minimum of 70 mm of asphalt cover was provided. It was predicted that the NI Good material would require 60 mm of asphalt cover.