FASTlab Radiosynthesis of the ¹⁸F-labelled HER2-binding Affibody molecule [¹⁸F]GE-226

An 18F‐labelled human epidermal growth factor receptor (HER2) receptor binding radiotracer is a potential tool to non‐invasively identify HER2 positive tumour lesions in subjects with recurrent metastatic breast cancer. Having explored the manual radiochemistry to conjugate the Affibody molecule ZHER2:2891 with [18F]4‐fluorobenzaldehyde, we have developed and optimised a full protocol for the automated GE FASTlab synthesiser. Our chemometric model predicted the best radiochemical purity for a short conjugation time (2.8 minutes), a low temperature (65°C), and a medium Affibody molecule precursor amount (5.5 mg). Under these optimised conditions, [18F]GE‐226 was produced after solid‐phase extraction purification with activity yield of 30% ± 7 (n = 18) and a radiochemical purity of 94% ± 2 (n = 18). The synthesis and purification was complete after 43 minutes and provided apparent molar activities of 12 to 30 GBq/μmol (n = 12) at the end of synthesis

Differential genetic interactions of yeast stress response MAPK pathways.

Genetic interaction screens have been applied with great success in several organisms to study gene function and the genetic architecture of the cell. However, most studies have been performed under optimal growth conditions even though many functional interactions are known to occur under specific cellular conditions. In this study, we have performed a large-scale genetic interaction analysis in Saccharomyces cerevisiae involving approximately 49 × 1,200 double mutants in the presence of five different stress conditions, including osmotic, oxidative and cell wall-altering stresses. This resulted in the generation of a differential E-MAP (or dE-MAP) comprising over 250,000 measurements of conditional interactions. We found an extensive number of conditional genetic interactions that recapitulate known stress-specific functional associations. Furthermore, we have also uncovered previously unrecognized roles involving the phosphatase regulator Bud14, the histone methylation complex COMPASS and membrane trafficking complexes in modulating the cell wall integrity pathway. Finally, the osmotic stress differential genetic interactions showed enrichment for genes coding for proteins with conditional changes in phosphorylation but not for genes with conditional changes in gene expression. This suggests that conditional genetic interactions are a powerful tool to dissect the functional importance of the different response mechanisms of the cell

High-throughput, quantitative analyses of genetic interactions in E. coli.

Large-scale genetic interaction studies provide the basis for defining gene function and pathway architecture. Recent advances in the ability to generate double mutants en masse in Saccharomyces cerevisiae have dramatically accelerated the acquisition of genetic interaction information and the biological inferences that follow. Here we describe a method based on F factor-driven conjugation, which allows for high-throughput generation of double mutants in Escherichia coli. This method, termed genetic interaction analysis technology for E. coli (GIANT-coli), permits us to systematically generate and array double-mutant cells on solid media in high-density arrays. We show that colony size provides a robust and quantitative output of cellular fitness and that GIANT-coli can recapitulate known synthetic interactions and identify previously unidentified negative (synthetic sickness or lethality) and positive (suppressive or epistatic) relationships. Finally, we describe a complementary strategy for genome-wide suppressor-mutant identification. Together, these methods permit rapid, large-scale genetic interaction studies in E. coli

Comedy in Unfunny Times: News Parody and Carnival after 9/11

Comedy has a special role in helping societies manage crisis moments, and the U.S. media paid considerable attention to the proper role of comedy in public culture after the 9/11 tragedies. As has been well documented, many popular U.S. comic voices were paralyzed in trying to respond to 9/11 or disciplined by audiences when they did. Starting with these obstacles in mind, this essay analyzes early comic responses to 9/11, and particularly those of the print and online news parody The Onion, as an example of how “fake” news discourse could surmount the rhetorical chill that fell over public culture after the tragedies. By exposing the news as “mere” production and by setting an agenda for learning about Islamic culture and Middle East politics, The Onion avoided violating decorum and invited citizen participation. This kind of meta-discourse was crucial after 9/11, when shifting rules for decorum created controversy and as official voices in government and media honed frames and narratives for talking about the attacks

Genetic interaction mapping informs integrative structure determination of protein complexes

Determining structures of protein complexes is crucial for understanding cellular functions. Here, we describe an integrative structure determination approach that relies on in vivo measurements of genetic interactions. We construct phenotypic profiles for point mutations crossed against gene deletions or exposed to environmental perturbations, followed by converting similarities between two profiles into an upper bound on the distance between the mutated residues. We determine the structure of the yeast histone H3-H4 complex based on similar to 500,000 genetic interactions of 350 mutants. We then apply the method to subunits Rpb1-Rpb2 of yeast RNA polymerase II and subunits RpoB-RpoC of bacterial RNA polymerase. The accuracy is comparable to that based on chemical cross-links; using restraints from both genetic interactions and cross-links further improves model accuracy and precision. The approach provides an efficient means to augment integrative structure determination with in vivo observations

Growth of microalgae with increased calorific values in a tubular bioreactor

In order to use microalgae as a fuel the algae should be of high calorific value and must be capable of growth in large volumes. Chlorella vulgaris and C. emersonii have been shown to grow in a 230 1 pumped tubular photobioreactor in Watanabe's medium and a low nitrogen medium. The low nitrogen medium induces higher lipid accumulation in both algae, which increased their calorific value. The highest calorific value was obtained with C. vulgaris (28 kJ g-1) grown in low nitrogen medium. However, the biomass productivity was 24 mg dry wt l-1 d-1 in the low nitrogen medium which was lower than in Watanabe's medium (40 mg dry wt l-1 d-1) and represents a reduced energy recovery. © 2002 Elsevier Science Ltd. All rights reserved