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The preparation of carbons for technical applications is typically based on a treatment of a precursor, which is transformed into the carbon phase with the desired structural properties. During such treatment the material passes through several different structural stages, for example, starting from precursor molecules via an amorphous phase into crystalline-like phases. While the structure of non-graphitic and graphitic carbon has been well studied, the transformation stages from molecular to amorphous and non-graphitic carbon are still not fully understood. Disordered carbon often contains a mixture of sp3-, sp2-and sp1-hybridized bonds, whose analysis is difficult to interpret. We systematically address this issue by studying the transformation of purely sp3-hybridized carbons, that is, nanodiamond and adamantane, into sp2-hybridized non-graphitic and graphitic carbon. The precursor materials are thermally treated at different temperatures and the transformation stages are monitored. We employ Raman spectroscopy, WAXS and TEM to characterize the structural changes. We correlate the intensities and positions of the Raman bands with the lateral crystallite size La estimated by WAXS analysis. The behavior of the D and G Raman bands characteristic for sp2-type material formed by transforming the sp3-hybridized precursors into non-graphitic and graphitic carbon agrees well with that observed using sp2-structured precursors.
NADP(H) is a central metabolic hub providing reducing equivalents to multiple biosynthetic, regulatory and antioxidative pathways in all living organisms. While biosensors are available to determine NADP+ or NADPH levels in vivo, no probe exists to estimate the NADP(H) redox status, a determinant of the cell energy availability. We describe herein the design and characterization of a genetically-encoded ratiometric biosensor, termed NERNST, able to interact with NADP(H) and estimate ENADP(H). NERNST consists of a redox-sensitive green fluorescent protein (roGFP2) fused to an NADPH-thioredoxin reductase C module which selectively monitors NADP(H) redox states via oxido-reduction of the roGFP2 moiety. NERNST is functional in bacterial, plant and animal cells, and organelles such as chloroplasts and mitochondria. Using NERNST, we monitor NADP(H) dynamics during bacterial growth, environmental stresses in plants, metabolic challenges to mammalian cells, and wounding in zebrafish. NERNST estimates the NADP(H) redox poise in living organisms, with various potential applications in biochemical, biotechnological and biomedical research.
