Mechanical and chemical unfolding of a single protein: A comparison

Is the mechanical unraveling of protein domains by atomic force microscopy (AFM) just a technological feat or a true measurement of their unfolding? By engineering a protein made of tandem repeats of identical Ig modules, we were able to get explicit AFM data on the unfolding rate of a single protein domain that can be accurately extrapolated to zero force. We compare this with chemical unfolding rates for untethered modules extrapolated to 0 M denaturant. The unfolding rates obtained by the two methods are the same. Furthermore, the transition state for unfolding appears at the same position on the folding pathway when assessed by either method. These results indicate that mechanical unfolding of a single protein by AFM does indeed reflect the same event that is observed in traditional unfolding experiments. The way is now open for the extensive use of AFM to measure folding reactions at the single-molecule level. Single-molecule AFM recordings have the added advantage that they define the reaction coordinate and expose rare unfolding events that cannot be observed in the absence of chemical denaturants.

Dynamic and quantitative Ca2+ measurements using improved cameleons

Cameleons are genetically-encoded fluorescent indicators for Ca2+ based on green fluorescent protein variants and calmodulin (CaM). Because cameleons can be targeted genetically and imaged by one- or two-photon excitation microscopy, they offer great promise for monitoring Ca2+ in whole organisms, tissues, organelles, and submicroscopic environments in which measurements were previously impossible. However, the original cameleons suffered from significant pH interference, and their Ca2+-buffering and cross-reactivity with endogenous CaM signaling pathways was uncharacterized. We have now greatly reduced the pH-sensitivity of the cameleons by introducing mutations V68L and Q69K into the acceptor yellow green fluorescent protein. The resulting new cameleons permit Ca2+ measurements despite significant cytosolic acidification. When Ca2+ is elevated, the CaM and CaM-binding peptide fused together in a cameleon predominantly interact with each other rather than with free CaM and CaM-dependent enzymes. Therefore, if cameleons are overexpressed, the primary effect is likely to be the unavoidable increase in Ca2+ buffering rather than specific perturbation of CaM-dependent signaling.

Avaliação dos impactos de políticas públicas de transferência de renda na qualidade de vida no semiárido nordestino face às mudanças climáticas

Introdução: Os impactos das alterações climáticas são e serão sentidos localmente, e irão variar de lugar para lugar. Nas regiões mais sensíveis à variabilidade climática, onde outros fatores e restrições afetam o desenvolvimento, a mudança climática terá um impacto significativamente maior. Uma da preocupações mais recentes da comunidade científica é que esses impactos negativos das alterações climáticas sobre a sociedade, saúde e o meio ambiente possa vir a restringir quaisquer ganhos realizados até o momento na melhora da qualidade de vida das populações, e em especial na região nordeste do Brasil. Neste contexto, as mudanças climáticas, bem como a variabilidade climática natural com seus extremos, podem vir a acentuar uma vulnerabilidade social já existente, fazendo com que a qualidade de vida das populações mais vulneráveis seja especialmente e desproporcionalmente afetada. Objetivo: Analisar como os impactos das políticas públicas de transferência de renda em especial o Programa Bolsa Família e a Aposentadoria Rural influênciam a qualidade de vida frente à variabilidade e mudanças climáticas de famílias pobres no nordeste brasileiro. Métodos: Fez-se uso de métodos qualitativos e quantitativos de coleta e análise de dados sobre as diferentes formas de pobreza e vulnerabilidade e a relação com a capacidade de adaptação à seca, e como isso afeta a percepção de qualidade de vida em famílias pobres rurais e urbanas em dois municípios do estado do Ceará (Parambu e Boa Viagem) sobre o estresse da seca. Realizou-se 221 entrevistas domiciliares (141 na zona rural e 80 na zona urbana). Complementarmente foram coletados dados qualitativos obtidos através de entrevistas semiestruturadas com informantes-chaves (gestores municipais e estaduais, chefes religiosos, presidentes de associações rurais e agentes de saúde). Os dados foram codificados e analisados com o auxílio do N-Vivo e do SPSS. Resultados: Os resultados foram separados e apresentados em formato de três manuscritos. Em geral, os resultados mostram que apesar dos programas de transferência de renda agirem positivamente na 16 melhora da qualidade de vida da famílias beneficiadas, durante o evento de seca as famílias não conseguem superar os fatores que as tornam vulneráveis, o que pode afetar sua percepção de bem-estar subjetivo. Ao mesmo tempo, devido às limitações e deficiências das intervenções políticas de intervenção de seca, as relações de dependência e clientelismo, característica da região, continuam a perpetuar. Conclusão: Os ganhos materiais na qualidade de vida, promovidos pelos programas sociais, como o Bolsa Família e a pensão rural, apesar de serem necessários, não são suficientes para promover uma transformação na vida dessas famílias

Involving Immigrants in Youth Sport Coaching: Part 1. A Literature Review and Quantitative Profile

Little, if any, previous research has investigated the involvement of immigrants in sport coaching. This is the first of a series of two articles that focus on this issue. In this article, following a general literature review, structured retrospective interviews (n = 29) are used to construct a profile of immigrant youth sport coaches. Two distinct profiles were identified: (a) leisure-oriented coaches, who had not coached prior to immigration; and (b) career-oriented coaches, who had coached prior to immigration Statistically significant differences (p < .05) were found between groups relating to athletic experience, sport-related education, time between immigration and initiation of coaching, and coaching of their own children Separate pathways to coaching involvement after immigration were identified for both profiles.

Application of the Hidden Markov Model for Innovative Projects "Viability" Analysis

This master thesis deals with determining of innovative projects "viability". "Viability" is the probability of innovative project being implemented. Hidden Markov Models are used for evaluation of this factor. The problem of determining parameters of model, which produce given data sequence with the highest probability, are solving in this research. Data about innovative projects contained in reports of Russian programs "UMNIK", "START" and additional data obtained during study are used as input data for determining of model parameters. The Baum-Welch algorithm which is one implementation of expectation-maximization algorithm is used at this research for calculating model parameters. At the end part of the master thesis mathematical basics for practical implementation are given (in particular mathematical description of the algorithm and implementation methods for Markov models).

Philosophical collections : containing an account of such physical, anatomical, chymical, mechanical, astronomical, optical, or other mathematical and philosophical experiments and observations as have lately come to the publishers hands, as also an account of some books of this kind lately published.

Mode of access: Internet.

Mathematics, from the points of view of the mathematician and of the physicist. An address delivered to the Mathematical and Physical Society of University College, London,

Cover title.

Mathematical and physical papers,

Vol III. Elasticity, heat, electro-magnetism; IV. Hydrodynamics and general dynamics; V. Thermodynamics, cosmical and geological physics, molecular and crystalline theory, electrodynamics; VI. Voltaic theory, radioactivity, electrons, navigation and tides, miscellaneous.

Plant analysis: qualitative and quantitative /

Mode of access: Internet.

Recent developments in physical chemistry : 3rd Mexican Meeting on Mathematical and Experimental Physics - México City, México - 10-14 September 2007 /

Mode of access: Internet.

Qualitative and quantitative determination of toxic materials in water supplies /

Mode of access: Internet.