Evidence-Based PET for Abdominal and Pelvic Tumours

Evidence-based data about the usefulness of positron emission tomography (PET) and hybrid imaging methods (PET/CT and PET/MRI) in abdominal and pelvic tumours have been collected and discussed in this chapter. These data were divided in three sections: (1) gastrointestinal tumours, (2) uro-genital tumours, (3) gynaecological tumours. Several pooled data (diagnostic and prognostic data), clinical settings (e.g. staging, restaging, treatment evaluation) and radiotracers as fluorine-18 fluorodeoxyglucose (18F-FDG), radiolabelled choline and prostate-specific membrane antigen (PSMA) were considered

Gene to Diagnostic: Self immobilizing protein for silica microparticle biosensor, modelled with sarcosine oxidase.

A rational design approach is proposed for a multifunctional enzyme reagent for point-of-care diagnostics. The biomaterial reduces downstream isolation steps and eliminates immobilization coupling chemicals for integration in a diagnostic platform. Fusion con-structs combined the central functional assay protein (e.g. monomeric sarcosine oxidase, mSOx, horseradish peroxidase, HRP), a visualizing protein (e.g. mCherry) and an in-built immobilization peptide (e.g. R5). Monitoring protein expression in E.coli was facilitated by following the increase in mCherry fluorescence, which could be matched to a color card, indicating when good protein expression has occurred. The R5 peptide (SSKKSGSYSGSKGSKRRIL) provided inbuilt affinity for silica and an immobilization capability for a silica based diagnostic, without requiring additional chemical coupling reagents. Silica particles extracted from beach sand were used to collect protein from crude protein extract with 85-95% selective uptake. The silica immobilized R5 pro-teins were stable for more than 2 months at room temperature. The Km for the silica-R52-mCh-mSOx-R5-6H was 16.5±0.9mM (com-pared with 16.5±0.4 mM, 16.3±0.3 mM, and 16.1±0.4 mM for R52-mCh-mSOx-R5-6H, mSOx-R5-6H and mSOx-6H respectively in solution). The use of the “silica-enzymes” in sarcosine and peroxide assays was shown, and a design using particle sedimentation through the sample was examined. Using shadowgraphy and particle image velocimetry the particle trajectory through the sample was mapped and an hourglass design with a narrow waist shown to give good control of particle position. The hourglass biosensor was demonstrated for sarcosine assay in the clinically useful range of 2.5 to 10 µM in both a dynamic and end point measurement regime

Research data supporting "Gene to Diagnostic: Self immobilizing protein for silica microparticle biosensor, modelled with sarcosine oxidase"

Analytical data supporting manuscript, including fluorescent data, laser scattering, protein sequence