Insights into DNA platination within unusual structural settings

2D [1H, 15N] HSQC NMR spectroscopy has been used to monitor reaction and product formation between [Pt(15NH3)2I2] and nucleic acids possessing irregular topologies and containing site specific phosphorothioate substitution in the phosphodiester backbone. Comparison of the reaction profiles of dimer nucleic acids with and without phosphorothioate substitution is made with their short nucleic acid counterparts containing the key dimer components. Whereas d(GpA) is relatively unreactive towards [Pt(15NH3)2I2], NMR evidence suggests that the tandem sheared mismatch duplex d(GCG3pAGC)2 reacts to form the head-to-tail inter-strand G3-N7-Pt-G3-N7 cross-link. The equivalent phosphorothioate R,S-d(GsA) reacts to form a mono-iodo, mono-sulphur adduct, whereas the tandem sheared mismatch phosphorothioate duplex d(GCGsAG5C)2 (VIs) reacts to form the unusual intra-strand macrochelate [Pt(15NH3)2{d(VIs-G5-N7)},S]2+ in which platinum is attached at both sulphur and G5-N7. Experimental evidence supports the formation of a stabilized mismatch duplex in which platinum is attached to two nitrogen centres in the sequence d(CGCGpTGCG) in contrast to R,S-d(CGCGsT5GCG) for which NMR evidence supports macrochelate-stabilized hairpin loop formation cross-linked at both phosphorothioate sulphur and T5-N

Diisopropylamide and TMP turbo-grignard reagents : a structural rationale for their contrasting reactivities

A neutral dimeric molecule in crystal form, the diisopropylamido turbo-Grignard reagent "(iPr2N)MgCl⋅LiCl" (see structure; blue N, red O, green Mg, yellow Cl, black C) separates into several charged ate species in dynamic exchange with each other in THF solution as determined by a combination of EXSY and DOSY NMR studies

Standardless, automated determination of Chlorine-35 by 35Cl nuclear magnetic resonance

A robust, fully automated, walk-up method is reported to quantify chloride in samples using 35Cl nuclear magnetic resonance. Minimal user input is required, no standards are acquired at the time of analysis; and the submission, acquisition, processing, and production of results are seamlessly integrated within existing software. The method demonstrated good linearity with R2 = 0.999 over three orders of magnitude of analyte concentration. The results were highly independent of analyte functionality, and the stability of instrument response was sufficient that analyses of additional standards were not required for a period of several months. At a nominal sample concentration of 10 mg/ml in D2O at 400 MHz, detection and quantitation limits of 0.1 and 0.5% (w/w) were achieved in a 1-h analysis time. Robust methodology was achieved by applying a rigorous approach to method development and validation to determine and evaluate fully the time- and sample-dependent factors that affect quantitation in these measurements

NMR analysis of Nile Blue (C. I. Basic Blue 12) and Thionine (C. I. 52000) in solution

The dyes Nile Blue (C. I. Basic Blue 12, NB) and Thionine (C. I. 52000, TH) were examined in both ionic and neutral forms in different solvents using NMR and UV-visible Spectroscopy to firmly establish the structures of the molecules and to assess the nature and extent of their aggregating characteristics. 1H and 13C NMR assignments and chemical shift data were used along with (for NB) nuclear Overhauser effect information enabling a structure for self-assembly to be proposed. In both cases these data were supplemented with variable temperature, dilution and diffusion-based experimental results using 1H NMR spectroscopy thereby enabling the extended aggregate structures to be assessed in terms of the relative strength of self-association and the extent to which extended aggregates could form. Full and detailed solution phase NMR analysis of such dyes, especially the two studied in this context, have either not been widely reported or have not been studied to the depth presented here. The data and their interpretation form an important addition to the analysis of this class of dye compounds and provide additional insight into the effects of self-assembly on the behaviour of such molecules in various solution-phase environments

Effects of motives on reactions to safe sun messages

We investigated whether appearance motive for sun exposure, which strongly predicts exposure behaviour, would predict reactions to safe sun messages. In a survey with an embedded experiment, 245 individuals completed measures of motives, read a safe sun message framed by incentive (appearance/health), tone (directive/nondirective) and valence (gain/loss), then completed measures of reactions. For participants high in appearance motive, an appearance-nondirective message was most persuasive. Regardless of individual's appearance motive, appearance messages produced lower reactance if phrased using nondirective language. To maximise persuasion and minimise reactance in individuals most motivated to sun expose, safe sun messages should focus on appearance using nondirective language

Closing the gap:how psychological distance influences willingness to engage in risky COVID behavior

Pandemics, and other risk-related contexts, require dynamic changes in behavior as situations develop. Human behavior is influenced by both explicit (cognitive) and implicit (intuitive) factors. In this study, we used psychological distance as a lens to understand what influences our decision-making with regard to risk in the context of COVID-19. This study was based on the rationale that our relational needs are more concrete to us than the risk of the virus. First, we explored the impact of social–psychological distance on participants’ risk perceptions and behavioral willingness. As hypothesized, we found that close social relationships of agents promoted willingness to engage in risky behavior. In the second phase, we tested an intervention designed to increase the concreteness of information about virus transmission as a mechanism to mitigate the bias of social influence. We found that the concreteness intervention resulted in significantly reduced willingness to engage in risky behavior. As such, communications aimed at changing the behavior of citizens during times of increased risk or danger should consider conceptually concrete messaging when communicating complex risk, and hence may provide a valuable tool in promoting health-related behavior

Structure and dynamics of the E. coli chemotaxis core signaling complex by cryo-electron tomography and molecular simulations

To enable the processing of chemical gradients, chemotactic bacteria possess large arrays of transmembrane chemoreceptors, the histidine kinase CheA, and the adaptor protein CheW, organized as coupled core-signaling units (CSU). Despite decades of study, important questions surrounding the molecular mechanisms of sensory signal transduction remain unresolved, owing especially to the lack of a high-resolution CSU structure. Here, we use cryo-electron tomography and sub-tomogram averaging to determine a structure of the Escherichia coli CSU at sub-nanometer resolution. Based on our experimental data, we use molecular simulations to construct an atomistic model of the CSU, enabling a detailed characterization of CheA conformational dynamics in its native structural context. We identify multiple, distinct conformations of the critical P4 domain as well as asymmetries in the localization of the P3 bundle, offering several novel insights into the CheA signaling mechanism

A detailed binding free energy study of 2 : 1 ligand–DNA complex formation by experiment and simulation

In 2004, we used NMR to solve the structure of the minor groove binder thiazotropsin A bound in a 2 : 1 complex to the DNA duplex, d(CGACTAGTCG)2. In this current work, we have combined theory and experiment to confirm the binding thermodynamics of this system. Molecular dynamics simulations that use polarizable or non-polarizable force fields with single and separate trajectory approaches have been used to explore complexation at the molecular level. We have shown that the binding process invokes large conformational changes in both the receptor and ligand, which is reflected by large adaptation energies. This is compensated for by the net binding free energy, which is enthalpy driven and entropically opposed. Such a conformational change upon binding directly impacts on how the process must be simulated in order to yield accurate results. Our MM-PBSA binding calculations from snapshots obtained from MD simulations of the polarizable force field using separate trajectories yield an absolute binding free energy (-15.4 kcal mol-1) very close to that determined by isothermal titration calorimetry (-10.2 kcal mol-1). Analysis of the major energy components reveals that favorable non-bonded van der Waals and electrostatic interactions contribute predominantly to the enthalpy term, whilst the unfavorable entropy appears to be driven by stabilization of the complex and the associated loss of conformational freedom. Our results have led to a deeper understanding of the nature of side-by-side minor groove ligand binding, which has significant implications for structure-based ligand development

Aerodynamic and Hydrodynamic Tests of a Family of Models of Flying Hulls Derived from a Streamline Body -- NACA Model 84 Series

A series of related forms of flying-boat hulls representing various degrees of compromise between aerodynamic and hydrodynamic requirements was tested in Langley Tank No. 1 and in the Langley 8-foot high-speed tunnel. The purpose of the investigation was to provide information regarding the penalties in water performance resulting from further aerodynamic refinement and, as a corollary, to provide information regarding the penalties in range or payload resulting from the retention of certain desirable hydrodynamic characteristics. The information should form a basis for over-all improvements in hull form