'Savoy Truffle': Love, Lust and Longing in a Box of Chocolates

This article re-reads The Beatles’ song, ‘Savoy Truffle’, not as an ode to Eric Clapton’s rotting teeth and chocolate consumption, but rather as a thinly veiled rock music metaphor reflecting the triptych love relationship between its composer, George Harrison, his wife, Pattie Boyd, and her lover and later husband, Eric Clapton. Re-reading ‘Savoy Truffle’ provides a valuable insight into the intricacies of how popular rock music communicates constructs of love within metaphor and how The Beatles integrated multiple meanings into lyrics conveying love and its contention. Such multiplicity aids explanations exploring the band’s extraordinary popularity by providing a socio-temporal insight into an extra-ordinary time: the 1960s. In re-reading ‘Savoy Truffle’, this article contextualizes the multiplicity of love itself within an exploration of the passion within lust, the longing of desire and the satisfaction of having the desired object. The desired object was Pattie Boyd, who Harrison projected within a box of ‘Good News’ chocolates

Recombinant human G6PD for quality control and quality assurance of novel point-of-care diagnostics for G6PD deficiency.

BACKGROUND:A large gap for the support of point-of-care testing is the availability of reagents to support quality control (QC) of diagnostic assays along the supply chain from the manufacturer to the end user. While reagents and systems exist to support QC of laboratory screening tests for glucose-6-phosphate dehydrogenase (G6PD) deficiency, they are not configured appropriately to support point-of-care testing. The feasibility of using lyophilized recombinant human G6PD as a QC reagent in novel point-of-care tests for G6PD deficiency is demonstrated. METHODS:Human recombinant G6PD (r-G6PD) was expressed in Escherichia coli and purified. Aliquots were stored at -80°C. Prior to lyophilization, aliquots were thawed, and three concentrations of r-G6PD (representing normal, intermediate, and deficient clinical G6PD levels) were prepared and mixed with a protective formulation, which protects the enzyme activity against degradation from denaturation during the lyophilization process. Following lyophilization, individual single-use tubes of lyophilized r-G6PD were placed in individual packs with desiccants and stored at five temperatures for one year. An enzyme assay for G6PD activity was used to ascertain the stability of r-G6PD activity while stored at different temperatures. RESULTS:Lyophilized r-G6PD is stable and can be used as a control indicator. Results presented here show that G6PD activity is stable for at least 365 days when stored at -80°C, 4°C, 30°C, and 45°C. When stored at 55°C, enzyme activity was found to be stable only through day 28. CONCLUSIONS:Lyophilized r-G6PD enzyme is stable and can be used as a control for point-of-care tests for G6PD deficiency

Performance of human recombinant glucose-6-phosphate dehydrogenase (r-G6PD) controls on qualitative tests for G6PD.

<p>Two lots of three concentrations (normal, intermediate, and deficient) of human r-G6PD were tested on two qualitative assays for G6PD: the fluorescent spot test (panels A and B) and a novel prototype rapid diagnostic test for G6PD (panels C and D). In panels A and B, the top row represents the Trinity normal controls for reference; the second, third, and fourth rows show the signals for the normal, intermediate, and deficient human r-G6PD controls, respectively. In panels C and D, the line intensity of test output correlates with expected enzyme activity, referring to normal, intermediate, and deficient activity. The signal for one lot of freshly lyophilized human r-G6PD (A and C) is compared to a second lot that had been stored at 4°C for more than one year (B and D).</p

Comparison of freshly lyophilized recombinant glucose-6-phosphate dehydrogenase (r-G6PD) versus year-old lyophilized r-G6PD and control.

<p>Human r-G6PD was lyophilized and assayed as “fresh” (grey bars) and compared to lyophilized r-G6PD samples from a lot that had been stored at 4°C for one year (striped bars). The Trinity controls were run at the same time for comparison of activity levels (black bars).</p

Stability of lyophilized human recombinant glucose-6-phosphate dehydrogenase (r-G6PD) reagent panel: Normal (blue), intermediate (green), and deficient (red).

<p>Lyophilized panel of three concentrations of human r-G6PD were stored for 12 months at five different temperatures: -80°C filled squares, 4°C filled circles, 30°C filled diamonds, 45°C filled triangles, and 55°C crosses. Activity was measured using the Trinity Biotech quantitative assay. The graph shows the stability of the panel over time. Stability starts to decline after approximately one month at a high temperature of 55°C. Thresholds are shown to indicate whether each measurement is still within an acceptable range of G6PD enzyme activity based on the ranges of Trinity controls: lower acceptable normal (blue dotted line), upper intermediate (black dotted line), lower intermediate (green dotted line), and upper deficient (red dotted line).</p