Botulinum neurotoxin type-A enters a non-recycling pool of synaptic vesicles

Neuronal communication relies on synaptic vesicles undergoing regulated exocytosis and recycling for multiple rounds of fusion. Whether all synaptic vesicles have identical protein content has been challenged, suggesting that their recycling ability may differ greatly. Botulinum neurotoxin type-A (BoNT/A) is a highly potent neurotoxin that is internalized in synaptic vesicles at motor nerve terminals and induces flaccid paralysis. Recently, BoNT/A was also shown to undergo retrograde transport, suggesting it might enter a specific pool of synaptic vesicles with a retrograde trafficking fate. Using high-resolution microscopy techniques including electron microscopy and single molecule imaging, we found that the BoNT/A binding domain is internalized within a subset of vesicles that only partially co-localize with cholera toxin B-subunit and have markedly reduced VAMP2 immunoreactivity. Synaptic vesicles loaded with pHrodo-BoNT/A-Hc exhibited a significantly reduced ability to fuse with the plasma membrane in mouse hippocampal nerve terminals when compared with pHrodo-dextran-containing synaptic vesicles and pHrodo-labeled anti-GFP nanobodies bound to VAMP2-pHluorin or vGlut-pHluorin. Similar results were also obtained at the amphibian neuromuscular junction. These results reveal that BoNT/A is internalized in a subpopulation of synaptic vesicles that are not destined to recycle, highlighting the existence of significant molecular and functional heterogeneity between synaptic vesicles.

Vacancy-Engineered Nanoceria: Enzyme Mimetic Hotspots for the Degradation of Nerve Agents

Organophosphorus-based nerve agents, such as paraoxon, parathion, and malathion, inhibit acetylcholinesterase, which results in paralysis, respiratory failure, and death. Bacteria are known to use the enzyme phosphotriesterase (PTE) to break down these compounds. In this work, we designed vacancy-engineered nanoceria (VE CeO2 NPs) as PTE mimetic hotspots for the rapid degradation of nerve agents. We observed that the hydrolytic effect of the nano-material is due to the synergistic activity between both Ce3+ and Ce4+ ions located in the active site-like hotspots. Furthermore, the catalysis by nanoceria overcomes the product inhibition generally observed for PTE and small molecule-based PTE mimetics.

Vacancy-Engineered Nanoceria: Enzyme Mimetic Hotspots for the Degradation of Nerve Agents

Organophosphorus-based nerve agents, such as paraoxon, parathion, and malathion, inhibit acetylcholinesterase, which results in paralysis, respiratory failure, and death. Bacteria are known to use the enzyme phosphotriesterase (PTE) to break down these compounds. In this work, we designed vacancy-engineered nanoceria (VE CeO2 NPs) as PTE mimetic hotspots for the rapid degradation of nerve agents. We observed that the hydrolytic effect of the nano-material is due to the synergistic activity between both Ce3+ and Ce4+ ions located in the active site-like hotspots. Furthermore, the catalysis by nanoceria overcomes the product inhibition generally observed for PTE and small molecule-based PTE mimetics.

Strain Rate-Dependent Compressive Deformation Behavior of Nd-Based Bulk Metallic Glass

Compressive deformation behavior of the Nd60Fe20Co10Al10 bulk metallic glass was characterized over a wide strain rate range (6.0 x 10(-4) to 1.0x10(3) s(-1)) at room temperature. Fracture stress was found to increase and fracture strain decrease with increasing applied strain rate. Serrated flow and a large number of shear bands were observed at the quasi-static strain rate (6.0 x 10(-4)s(-1)). The results suggest that the appearance of a large number of shear bands is probably associated with flow serration observed during compression; and both shear banding and flow serration are a strain accommodation and stress relaxation process. At dynamic strain rates (1.0 x 10(3) s(-1)), the rate of shear band nucleation is not sufficient to accommodate the applied strain rate and thus causes an early fracture of the test sample. The fracture behavior of the Nd60Fe20Co10Al10 bulk metallic glass is sensitive to strain rate.

Strain-Induced Grain Refinement of Cobalt During Surface Mechanical Attrition Treatment

The microstructural evolution during surface mechanical attrition treatment of cobalt (a mixture of hexagonal close packed (hep) and face-centered cubic (fcc) phases) was investigated. In order to reveal the mechanism of grain refinement and strain accommodation. The microstructure was systematically characterized by both cross-sectional and planar-view transmission electron microscopy. In the hcp phase, the process of grain refinement. Accompanied by an increase in strain imposed in the surface layer. Involved: (1) the onset of 110 111 deformation twinning, (2) the operation of (1 120) 110 1 0} prismatic and (1 120) (000 1) basal slip, leading to the formation of low-angle dislocation boundaries, and (3) the successive subdivision of grains to a finer and finer scale. Ressulting in the formation of highly misoriented nanocrystalline grains. Moreover. The formation of nanocrystalliies at the grain boundary and triple junction was also observed to occur concurrently with straining. By contrast. The fec phase accommodated strain in a sequence as follows: (1) slip of dislocations by forming intersecting planar arrays of dislocations, (2) {1 1 1} deformation twinning, and (3) the gamma(fcc) --> epsilon(hcp) martensitic phase transformation. The mechanism of grain refinement was interpreted in terms of the structural subdivision of grains together with dynamic recrystallization occurring in the hep phase and the gamma --> E: martensitic transformation in the fcc phase as well.

La gestión del turismo en los programas de innovación rural de Castilla-La Mancha

[ES] Los programas de innovación rural han supuesto un verdadero revulsivo para determinadas regiones europeas, por cuanto han representado una cuantiosa inversión que ha necesitado de una adecuada gestión para dirigir el destino de unos fondos estructurales. En este sentido, el presente trabajo analiza la inversión de estos programas de desarrollo en medidas de apoyo al sector del turismo rural, que han supuesto un importante incremento de la oferta a diversos niveles: aumento de alojamientos rurales, del número de plazas disponibles para turistas, rehabilitación del patrimonio, puesta en valor de elementos históricos y recuperación, en definitiva, del patrimonio histórico, cultural, arquitectónico y natural del mundo rural.

Interações entre o Legislativo e o Executivo federal do Brasil na definição de políticas de interesse amplo: uma abordagem sistêmica, com aplicação na saúde

Aborda a relação entre o Legislativo e o Executivo na produção de políticas. Identifica os elementos do sistema de produção legislativa do Brasil (regras estruturantes, atores, recursos, instâncias de decisão e tipos de políticas produzidas) e propõe um modelo para o caso brasileiro de presidencialismo de coalizão, com base em estudos sobre a relação entre o presidente e o Congresso dos EUA e também na vasta produção existente sobre o contexto nacional. O sistema é estruturado pelo marco normativo de maior hierarquia, a Constituição, determinado historicamente, o qual privilegia a governabilidade com "accountability" e também orienta políticas segundo princípios de equidade, mas com responsabilidade orçamentária. O modelo considera que as agendas estratégicas dos atores são produto de variadas influências, incluindo o ¿status quo¿ (políticas existentes) e as demandas provenientes das conexões normativa e eleitoral. A partir desse modelo, o estudo analisa seus elementos e relações, aplicando-o a um conjunto abrangente de propostas legislativas (cerca de 21 mil proposições sobre todos os temas, apresentadas no Congresso entre 1999 e 2006, nas três vias).

O Congresso Nacional e a política externa brasileira : posicionamento dos senadores frente às negociações para a formação da ALCA (1994-2005)

Analisa o posicionamento dos integrantes do Congresso Nacional frente às negociações para a formação da Área de Livre Comércio das Américas (ALCA), a concentrar sua atenção sobre o Senado Federal, com eventuais incursões pela Câmara dos Deputados. O período por nós delimitado compreende, respectivamente, o lançamento da proposta pela integração continental, ocorrida na I Cúpula das Américas, nos EUA, em 1994, até o ano de 2005, em que se evidenciou a paralisação do processo negociador. Dois fatores estimularam-nos a realizar esta tarefa. Primeiro, colocar à prova a tese, disseminada pelo senso comum, de que o Parlamento não demonstra interesse por questões ou assuntos internacionais. Segundo, fornecer uma contribuição à escassa bibliografia disponível a respeito da relação entre os congressistas e a produção da política externa brasileira de integração regional, em particular. Apesar de não ignorarmos o papel predominante do Poder Executivo na formulação e condução da política externa nacional, e nem a comprovada carência de mecanismos participativos e decisórios formais à disposição do Parlamento para atuar nas diferentes facetas deste plano, procuramos aferir em que medida a atividade parlamentar se restringe ou não, na prática, ao seu exercício constitucional de aprovar ou rejeitar matérias sobre política exterior, conforme nos aponta o senso comum. O caráter polêmico das negociações e dos debates ocorridos no Brasil a respeito da conformação da ALCA, ao longo de mais de uma década, levou-nos a questionar qual teria sido o posicionamento dos congressistas com relação ao assunto, ou seja, se teriam manifestado interesse em se envolver mais intensamente com a questão, ou se reservado a debatê-la somente na ocasião em que fosse submetida, na forma de um acordo, à sua apreciação. A elaboração do presente trabalho orientou-se pela pretensão em alcançar respostas para duas perguntas formuladas por nós: a ausência de mecanismos formais de participação parlamentar nas negociações sobre a ALCA teria refletido um suposto desinteresse dos senadores pela questão? Em que medida esta referida ausência não teria impelido o Parlamento a buscar influenciar o processo por outros meios, ou até mesmo fomentado demandas pela ampliação de seu papel constitucional no tratamento de questões externas? Para tentar responder a tais perguntas, servimo-nos especialmente dos apanhamentos taquigráficos dos pronunciamentos realizados pelos senadores; de proposições elaboradas pelos congressistas e de atas e notas tquigráficas de audiências públicas promovidas por Comissões do Congresso Nacional.

Unlike Physical Exercise, Modified Environment Increases the Lifespan of SOD1(G93A) Mice However Both Conditions Induce Cellular Changes

8 p.

Designing Spaces for Effective Learning: A guide to 21st century learning space design

Increasing investment in estate and learning technologies, combined with the need for more cost-effective space utilisation, is making it increasingly important for senior managers to keep abreast of new thinking about the design of technology-rich learning spaces. Designing Spaces for Effective Learning, one of a series of guides, was launched at the JISC Conference 2006 which helped to meet this need. A visually-rich publication, it was designed to promote better understanding of what makes an effective design for the 21 century and to summarise the key points to consider when approaching a refurbishment or new-build project. The publication takes the reader on a ’walk through’ an educational institution, exploring the relationship between learning technologies and innovative examples of physical space design at each stage of the journey. Discussion of the key points is illustrated by ten case studies from further and higher education, and floor plans from AMA Alexi Marmot Associates, architects and space planners, which provide up-to-date guidelines on the integration of technologies into teaching and learning accommodation.

Nearly free molecular heat transfer from a sphere

Consider a sphere immersed in a rarefied monatomic gas with zero mean flow. The distribution function of the molecules at infinity is chosen to be a Maxwellian. The boundary condition at the body is diffuse reflection with perfect accommodation to the surface temperature. The microscopic flow of particles about the sphere is modeled kinetically by the Boltzmann equation with the Krook collision term. Appropriate normalizations in the near and far fields lead to a perturbation solution of the problem, expanded in terms of the ratio of body diameter to mean free path (inverse Knudsen number). The distribution function is found directly in each region, and intermediate matching is demonstrated. The heat transfer from the sphere is then calculated as an integral over this distribution function in the inner region. Final results indicate that the heat transfer may at first increase over its free flow value before falling to the continuum level.

Dramatické texty přeložené a cenzurované ve frankistickém Španělsku (šedesátá léta): divadlo a film jako kontrolovaná představení

Texto en checo. Editora de la obra Petra Vavroušová.

Cooperative properties of lipid bilayers : collective director fluctuations and the effects of hydrophobic mismatch in protein/lipid membrane systems

Cooperative director fluctuations in lipid bilayers have been postulated for many years. ^2H-NMR T_1^(-1), T_(1P)^(-1) , and T_2^(-1); measurements have been used identify these motions and to determine the origin of increased slow bilayer motion upon addition of unlike lipids or proteins to a pure lipid bilayer.

The contribution of cooperative director fluctuations to NMR relaxation in lipid bilayers has been expressed mathematically using the approach of Doane et al.^1 and Pace and Chan.^2 The T_2^(-1)’s of pure dimyristoyllecithin (DML) bilayers deuterated at the 2, 9 and 10, and all positions on both lipid hydrocarbon chains have been measured. Several characteristics of these measurements indicate the presence of cooperative director fluctuations. First of all, T_2^(-1) exhibits a linear dependence on S2/CD. Secondly, T_2^(-1) varies across the ^2H-NMR powder pattern as sin^2 (2, β), where , β is the angle between the average bilayer director and the external magnetic field. Furthermore, these fluctuations are restricted near the lecithin head group suggesting that the head group does not participate in these motions but, rather, anchors the hydrocarbon chains in the bilayer.

T_2^(-1)has been measured for selectively deuterated liquid crystalline DML hilayers to which a host of other lipids and proteins have been added. The T_2^(-1) of the DML bilayer is found to increase drastically when chlorophyll a (chl a) and Gramicidin A' (GA') are added to the bilayer. Both these molecules interfere with the lecithin head group spacing in the bilayer. Molecules such as myristic acid, distearoyllecithin (DSL), phytol, and cholesterol, whose hydrocarbon regions are quite different from DML but which have small,neutral polar head groups, leave cooperative fluctuations in the DML bilayer unchanged.

The effect of chl a on cooperative fluctuations in the DML bilayer has been examined in detail using ^2H-NMR T_1^(-1), T_(1P)^(-1) , and T_2^(-1); measurements. Cooperative fluctuations have been modelled using the continuum theory of the nematic state of liquid crystals. Chl a is found to decrease both the correlation length and the elastic constants in the DML bilayer.

A mismatch between the hydrophobic length of a lipid bilayer and that of an added protein has also been found to change the cooperative properties of the lecithin bilayer. Hydrophobic mismatch has been studied in a series GA' / lecithin bilayers. The dependence of 2H-NMR order parameters and relaxation rates on GA' concentration has been measured in selectively deuterated DML, dipalmitoyllecithin (DPL), and DSL systems. Order parameters, cooperative lengths, and elastic constants of the DML bilayer are most disrupted by GA', while the DSL bilayer is the least perturbed by GA'. Thus, it is concluded that the hydrophobic length of GA' best matches that of the DSL bilayer. Preliminary Raman spectroscopy and Differential Scanning Calorimetry experiments of GA' /lecithin systems support this conclusion. Accommodation of hydrophobic mismatch is used to rationalize the absence of H_(II) phase formation in GA' /DML systems and the observation of H_(II) phase in GA' /DPL and GA' /DSL systems.

1. J. W. Doane and D. L. Johnson, Chem. Phy3. Lett., 6, 291-295 (1970). 2. R. J. Pace and S. I. Chan, J. Chem. Phy3., 16, 4217-4227 (1982).

Molecular genetic studies on voltage-gated ion channels

Several different methods have been employed in the study of voltage-gated ion channels. Electrophysiological studies on excitable cells in vertebrates and molluscs have shown that many different voltage-gated potassium (K⁺) channels and sodium channels may coexist in the same organism. Parallel genetic studies in Drosophila have identified mutations in several genes that alter the properties of specific subsets of physiologically identified ion channels. Chapter 2 describes molecular studies that identify two Drosophila homologs of vertebrate sodium-channel genes. Mutations in one of these Drosophila sodium-channel genes are shown to be responsible for the temperature-dependent paralysis of a behavioural mutant para^ts. Evolutionary arguments, based on the partial sequences of the two Drosophila genes, suggest that subfamilies of voltage-gated sodium channels in vertebrates remain to be identified.

In Drosophila, diverse voltage-gated K⁺ channels arise from alternatively spliced mRNAs generated at the Shaker locus. Chapter 3 and the Appendices describe the isolation and characterization of several human K⁺-channel genes, similar in sequence to Shaker. Each of these human genes has a highly conserved homolog in rodents; thus, this K⁺-channel gene family probably diversified prior to the mammalian radiation. Functional K⁺ channels encoded by these genes have been expressed in Xenopus oocytes and their properties have been analyzed by electrophysiological methods. These studies demonstrate that both transient and noninactivating voltage-gated K⁺ channels may be encoded by mammalian genes closely related to Shaker. In addition, results presented in Appendix 3 clearly demonstrate that independent gene products from two K⁺-channel genes may efficiently co-assemble into heterooligomeric K⁺ channels with properties distinct from either homomultimeric channel. This finding suggests yet another molecular mechanism for the generation of K⁺-channel diversity.

A photographic record of a pre- and post-lake creation event from World War II at Valley, Isle of Anglesey, UK

The article compares a recent aerial photograph of the lowlands of the Isle of Anglesey area with a German surveillance photograph from 1941. The authors aim to infer the environmental changes made to this sand dune and lake system as a direct consequence of constructing the airfield. Part of Tywyn Trewan, the extensive sand dune system, was completely destroyed in order to create runways and the technical and domestic accommodation to house a strategic airfield. As part of the dredging, six new water bodies with a combined surface area of approximately 6 ha were created.