Focused molecular analysis of small cell lung cancer: feasibility in routine clinical practice.

Background: There is an urgent need to identify molecular signatures in small cell lung cancer (SCLC) that may select patients who are likely to respond to molecularly targeted therapies. In this study, we investigate the feasibility of undertaking focused molecular analyses on routine diagnostic biopsies in patients with SCLC.

Methods: A series of histopathologically confirmed formalin-fixed, paraffin-embedded SCLC specimens were analysed for epidermal growth factor receptors (EGFR), KRAS, NRAS and BRAF mutations, ALK gene rearrangements and MET amplification. EGFR and KRAS mutation testing was evaluated using real time polymerase chain reaction (RT-PCR cobas®), BRAF and NRAS mutations using multiplex PCR and capillary electrophoresis-single strand conformation analysis, and ALK and MET aberrations with fluorescent in situ hybridization. All genetic aberrations detected were validated independently.

Results: A total of 105 patients diagnosed with SCLC between July 1990 and September 2006 were included. 60 (57 %) patients had suitable tumour tissue for molecular testing. 25 patients were successfully evaluated for all six pre-defined molecular aberrations. Eleven patients failed all molecular analysis. No mutations in EGFR, KRAS and NRAS were detected, and no ALK gene rearrangements or MET gene amplifications were identified. A V600E substitution in BRAF was detected in a Caucasian male smoker diagnosed with SCLC with squamoid and glandular features.

Conclusion: The paucity of patients with sufficient tumour tissue, quality of DNA extracted and low frequency of aberrations detected indicate that alternative molecular characterisation approaches are necessary, such as the use of circulating plasma DNA in patients with SCLC.

β-Strand mimetic foldamers rigidified through dipolar repulsion

Many therapeutically relevant protein-protein interactions contain hot-spot regions on secondary structural elements, which contribute disproportionately to binding enthalpy. Mimicry of such α-helical regions has met with considerable success, however the analogous approach for the β-strand has received less attention. Presented herein is a foldamer for strand mimicry in which dipolar repulsion is a central determinant of conformation. Computation as well as solution- and solid-phase data are consistent with an ensemble weighted almost exclusively in favor of the desired conformation.

Field investigation of contactless displacement measurement using computer vision systems for civil engineering applications

Much of the bridge stock on major transport links in North America and Europe was constructed in the 1950s and 1960s and has since deteriorated or is carrying loads far in excess of the original design loads. Structural Health Monitoring Systems (SHM) can provide valuable information on the bridge capacity but the application of such systems is currently limited by access and bridge type. This paper investigates the use of computer vision systems for SHM. A series of field tests have been carried out to test the accuracy of displacement measurements using contactless methods. A video image of each test was processed using a modified version of the optical flow tracking method to track displacement. These results have been validated with an established measurement method using linear variable differential transformers (LVDTs). The results obtained from the algorithm provided an accurate comparison with the validation measurements. The calculated displacements agree within 2% of the verified LVDT measurements, a number of post processing methods were then applied to attempt to reduce this error.

A Loop-mediated Isothermal Amplification Method for Rapid Direct Detection and Differentiation of Non-pathogenic and Verocytotoxigenic Escherichia coli in Beef and Bovine Faeces

The aim of this study was to develop a multiplex loop-mediated isothermal amplification (LAMP) method capable of detecting Escherichia coli generally and verocytotoxigenic E. coli (VTEC) specifically in beef and bovine faeces. The LAMP assay developed was highly specific (100%) and able to distinguish between E. coli and VTEC based on the amplification of the phoA, and stx1 and/or stx2 genes, respectively. In the absence of an enrichment step, the limit of detection 50% (LOD50) of the LAMP assay was determined to be 2.83, 3.17 and 2.83-3.17 log CFU/g for E. coli with phoA, stx1 and stx2 genes, respectively, when artificially inoculated minced beef and bovine faeces were tested. The LAMP calibration curves generated with pure cultures, and spiked beef and faeces, suggested that the assay had good quantification capability. Validation of the assay, performed using retail beef and bovine faeces samples, demonstrated good correlation between counts obtained by the LAMP assay and by a conventional culture method, but suggested the possibility of false negative LAMP results for 12.5-14.7% of samples tested. The multiplex LAMP assay developed potentially represents a rapid alternative to culture for monitoring E.coli levels in beef or faeces and it would provide additional information on the presence of VTEC. However, some further optimisation is needed to improve detection sensitivity.

Direct Field Measurement of the Dynamic Amplification in a Bridge

In this paper, the level of dynamics, as described by the Assessment Dynamic Ratio (ADR), is measured directly through a field test on a bridge in the United Kingdom. The bridge was instrumented using fiber optic strain sensors and piezo-polymer weigh-in-motion sensors were installed in the pavement on the approach road. Field measurements of static and static-plus-dynamic strains were taken over 45 days. The results show that, while dynamic amplification is large for many loading events, these tend not to be the critical events. ADR, the allowance that should be made for dynamics in an assessment of safety, is small.

Laboratory investigation of contactless displacement measurement using computer vision systems

Much of the bridge stock on major transport links in North America and Europe was constructed in the 1950’s and 1960’s and has since deteriorated or is carrying loads far in excess of the original design loads. Structural Health Monitoring Systems (SHM) can provide valuable information on the bridge capacity but the application of such systems is currently limited by access and system cost. This paper investigates the development of a low cost portable SHM system using commercially available cameras and computer vision techniques. A series of laboratory tests have been carried out to test the accuracy of displacement measurements using contactless methods. The results from each of the tests have been validated with established measurement methods, such as linear variable differential transformers (LVDTs). A video image of each test was processed using two different digital image correlation programs. The results obtained from the digital image correlation methods provided an accurate comparison with the validation measurements. The calculated displacements agree within 4% of the verified measurements LVDT measurements in most cases confirming the suitability full camera based SHM systems