Thermal dissociation of multimeric protein complexes by using nanoelectrospray mass spectrometry

The behavior of macromolecular systems at different temperatures is often crucial to their biological activity and function. While heat-induced changes of individual proteins are readily monitored by a number of spectroscopic methods, changes in noncovalent complexes of biomolecules are more challenging to interpret. Nanoelectrospray mass spectrometry is becoming increasingly powerful in the study of large noncovalent complexes, and here we describe the design, characterization, and application of a novel probe that allows the thermocontrol of the solution in the electrospray capillary. The transition temperature for the unfolding of the protein lysozyme is readily obtained and correlates closely with that measured by fluorescence spectroscopy, thereby demonstrating the validity of this approach. We apply this technique to the study of the 200-kDa complex of the small heat shock protein TaHSP16.9, revealing both its dissociation into suboligomeric species and an increase in its size and polydispersity at elevated temperatures. In contrast, gas-phase activation of this complex is also carried out and yields a dissociation pathway fundamentally different from that observed for thermal activation in solution. As such, this probe allows the study of the reversible heat-induced changes of noncovalent complexes in a biologically relevant manner.     

Use of a microchip device coupled with mass spectrometry for ligand screening of a multi-protein target

Nanoflow electrospray mass spectrometry has been applied previously to investigate noncovalent protein-protein and protein-ligand interactions. Here we evaluate a commercial microchip device for this application. We show that the microchip can be used to obtain mass spectra of the noncovalent tetramer transthyretin. The device showed a 10-fold increase in signal stability compared with a nanoflow capillary and a high level of nozzle-to-nozzle reproducibility. Binding of the natural ligand thyroxine was clearly observed, and a range of small molecules proposed as inhibitors of transthyretin amyloidosis were shown to be effective in stabilizing the tetramer. We propose that measuring the ability of small molecules to stabilize protein complexes using this automated microchip technology will enable high-throughput screening of multi-protein complexes by mass spectrometry.     

Environmental stress and recovery: the geochemical record of human disturbance in New Bedford Harbor and Apponagansett Bay,Massachusetts (USA)

Sediments record the history of contamination to estuaries. Analysis of the concentrations of toxic organic compounds, contaminant and crustal metals, organic carbon content and isotopic composition in sediment cores from two estuarine systems in Buzzards Bay allowed reconstruction of human impacts over 350 years. Vertical distributions of the contaminants correlate with changes in the nature of watershed/estuarine activities. All contaminants were highly enriched (tens to hundreds times background) in modern New Bedford Harbor sediments. Enrichment began around the turn of the 20th century for all but PCBs, which were first synthesized in the 1930s. An increase in organic carbon content and a shift of carbon isotopes toward a more terrestrial signature illustrates increasing anthropogenic impact in New Bedford as population grew along with the industrial base. Institution of environmental protection measures in the late 20th century was reflected in decreased, although still substantially elevated, concentrations of contaminants. A lack of industrial development in Apponagansett Bay resulted in much lower concentrations of the same indicators, although specific contaminants related to the early whaling industry increased significantly above background as early as the late 18th century. The similarity of indicators in older portions of cores from NBH and unimpacted Apponagansett Bay demonstrates that cores can be used to establish reference conditions as successfully as using separate sites judged a priori to represent the reference state. The historical reconstruction approach provides the basis for establishing relationships between environmental stressors and factors that drive the stressors, as well as a framework for the assessment of ecological response(s) to environmental stressors over a range of time and/or exposure scales.     

Knowledge, strategy, and motivational contributions to preschool children's object recall.

This study examined the contributions of four individual difference predictors to preschool children's recall for object names. Measures of object knowledge (the Peabody Picture Vocabulary Test [PPVT]), conceptual tempo (the Kansas Reflection-Impulsivity Scale for Preschoolers [KRISP] error scores), strategic study period activities, and teacher ratings of children's mastery motivation were used to predict recall scores of 3- and 4-year-old children. KRISP-error scores, strategic study activities, and motivation ratings were found to be significant correlates of recall. Motivation ratings entered a regression prediction equation first, followed by the strategic study activity measure, yielding a multiple correlation of .62. These predictors were unrelated, suggesting that motivational factors contributed to young children's recall proficiency directly, perhaps through greater task involvement and analysis and more effortful retrieval activity, and do not mediate strategic study behavior. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Thick Film Patterned Ceramics Using UV-Curable Pastes

Screen-printed layers of Al₂O₃, BaTiO₃, 0.90Pb(Mg_1/3Nb_2/3)-0₃–0.10PbTiO₃, Pb(Zr,Ti)O₃, ZnO, and glass alumina pastes have been patterned using photoprinting techniques. Curability of each paste by ultraviolet light and formation of holes with various diameters were studied for application as a production method for very small-sized vias. The dependence of hole diameters on various experimental parameters is presented and discussed. Wall slopes were found to depend on the transmission of the powders used in the pastes.     

Gender differences in factors predicting unsafe crossing decisions in adult pedestrians across the lifespan: A simulation study

Adult pedestrian accident data has demonstrated that the risk of being killed or seriously injured varies with age and gender. A range of factors affecting road crossing choices of 218 adults aged 17-90+ were examined in a simulation study using filmed real traffic. With increasing age, women were shown to make more unsafe crossing decisions, to leave small safety margins and to become poorer at estimating their walking speed. However, the age effects on all of these were ameliorated by driving experience. Men differed from women in that age was not a major factor in predicting unsafe crossing decisions. Rather, reduced mobility was the key factor, leading them to make more unsafe crossings and delay longer in leaving the kerb. For men, driving experience did not predict unsafe road crossing decisions. Although male drivers were more likely to look both ways before crossing than male non-drivers, the impact of being a driver had a negative effect in terms of smaller safety margins and delay in leaving the kerb. The implications of the different predictor variables for men and women for unsafe road crossing are discussed and possible reasons for the differences explored.