Recoupled separated-local-field experiments and applications to study intermediate-regime molecular motions

A specific separated-local-field NMR experiment, dubbed Dipolar-Chemical-Shift Correlation (DIPSHIFT) is frequently used to study molecular motions by probing reorientations through the changes in XH dipolar coupling and T-2. In systems where the coupling is weak or the reorientation angle is small, a recoupled variant of the DIPSHIFT experiment is applied, where the effective dipolar coupling is amplified by a REDOR-like pi-pulse train. However, a previously described constant-time variant of this experiment is not sensitive to the motion-induced T-2 effect, which precludes the observation of motions over a large range of rates ranging from hundreds of Hz to around a MHz. We present a DIPSHIFT implementation which amplifies the dipolar couplings and is still sensitive to T-2 effects. Spin dynamics simulations, analytical calculations and experiments demonstrate the sensitivity of the technique to molecular motions, and suggest the best experimental conditions to avoid imperfections. Furthermore, an in-depth theoretical analysis of the interplay of REDOR-like recoupling and proton decoupling based on Average-Hamiltonian Theory was performed, which allowed explaining the origin of many artifacts found in literature data. (C) 2012 Elsevier Inc. All rights reserved.

Slice: experiência de gestão de um processo de projeto paramétrico, com superfícies complexas

O artigo apresenta uma reflexão sobre a gestão do processo de projeto paramétrico, com superfícies complexas, baseado em duas experiências realizadas no Instituto de Arquitetura e Urbanismo, da USP de São Carlos, pelo grupo de pesquisa Nomads.usp. SLICE é o resultado parcial de um pavilhão temporário, com superfícies complexas, desenvolvido por meio de projeto paramétrico e técnicas de fabricação digital. Utilizou-se o procedimento de uma pesquisa-ação2 com alunos de graduação e recém-formados em arquitetura, arquitetos do Nomads.usp e a indústria. Os experimentos apontaram para reflexões sobre algumas mudanças na gestão destes processos, para facilitar a viabilidade técnico-construtiva e gerar conhecimento nas áreas de projeto paramétrico, fabricação digital e gestão. O artigo tem caráter exploratório e o desafio é entender e avaliar a gestão do projeto paramétrico em relação a restrição de material, custo de execução e tempo de produção.

Uma arqueologia de diagramas cibernéticos

This paper investigates the use of explicit structures of information in architectural design. Particularly, it approaches the use of diagrams related to cybernetics and information theory in experimental practices in the 1960’s and 1970’s. It analyses the diagram of cybernetic control proposed by the cybernetician Gordon Pask for the Fun Palace, the diagrams produced by the utopian architect Yona Friedman in the conceptual description of the Flatwriter program and Christopher Alexander’s diagrams and his theories of Synthesis of Form and Pattern Language. Finally it establishes a brief parallel between current domestication and use of dataflow programming with the cybernetic diagrams, highlighting differences in their complexity approach.

Endo-PDI is required for TNFα-induced angiogenesis

Protein disulfide isomerase (PDI) and its homologs are oxidoreductases facilitating protein folding in the ER. Endo-PDI (also termed ERp46) is highly expressed in endothelial cells. It belongs to the PDI family but its physiological function is largely unknown. We studied the role of Endo-PDI in endothelial angiogenic responses. Stimulation of human umbilical vein endothelial cells (with TNFα (10ng/ml) increased ERK1/2 phosphorylation. This effect was largely attenuated by Endo-PDI siRNA, whereas JNK and p38 MAP kinase phosphorylation was Endo-PDI independent. Similarly, TNFα-stimulated NF-κB signaling determined by IκBα degradation as well as TNFα-induced ICAM expression was unaffected by Endo-PDI siRNA. The action of Endo-PDI was not mediated by extracellular thiol exchange or cell surface PDI as demonstrated by nonpermeative inhibitors and PDI-neutralizing antibody. Moreover, exogenously added PDI failed to restore ERK1/2 activation after Endo-PDI knockdown. This suggests that Endo-PDI acts intracellularly potentially by maintaining the Ras/Raf/MEK/ERK pathway. Indeed, knockdown of Endo-PDI attenuated Ras activation measured by G-LISA and Raf phosphorylation. ERK activation influences gene expression by the transcriptional factor AP-1, which controls MMP-9 and cathepsin B, two proteases required for angiogenesis. TNFα-stimulated MMP-9 and cathepsin B induction was reduced by silencing of Endo-PDI. Accordingly, inhibition of cathepsin B or Endo-PDI siRNA blocked the TNFα-stimulated angiogenic response in the spheroid outgrowth assays. Moreover ex vivo tube formation and in vivo Matrigel angiogenesis in response to TNFα were attenuated by Endo-PDI siRNA. In conclusion, our study establishes Endo-PDI as a novel, important mediator of AP-1-driven gene expression and endothelial angiogenic function