21 resultados para Cytoplasmic filaments

em Cambridge University Engineering Department Publications Database


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A model to describe the cavitation-induced breakage of nanofilaments during their sonication in solution is proposed. The model predicts a limiting length below which scission no longer occurs, and accurately describes experimental results for materials ranging from carbon nanotubes to protein fibrils. Sonication-induced breakage can now be used as a probe for the strength of nanostructures. © 2009 WILEY-VCH Verlag GmbH & Co. KGaA.

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Self-assembly processes resulting in linear structures are often observed in molecular biology, and include the formation of functional filaments such as actin and tubulin, as well as generally dysfunctional ones such as amyloid aggregates. Although the basic kinetic equations describing these phenomena are well-established, it has proved to be challenging, due to their non-linear nature, to derive solutions to these equations except for special cases. The availability of general analytical solutions provides a route for determining the rates of molecular level processes from the analysis of macroscopic experimental measurements of the growth kinetics, in addition to the phenomenological parameters, such as lag times and maximal growth rates that are already obtainable from standard fitting procedures. We describe here an analytical approach based on fixed-point analysis, which provides self-consistent solutions for the growth of filamentous structures that can, in addition to elongation, undergo internal fracturing and monomer-dependent nucleation as mechanisms for generating new free ends acting as growth sites. Our results generalise the analytical expression for sigmoidal growth kinetics from the Oosawa theory for nucleated polymerisation to the case of fragmenting filaments. We determine the corresponding growth laws in closed form and derive from first principles a number of relationships which have been empirically established for the kinetics of the self-assembly of amyloid fibrils.

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The operation on how high quality single-mode operation can be readily attained on etching circles in multimode devices is discussed. Arrays of such spots can also be envisaged. Control of the polarization state is also achieved by use of deep line etches. The output filaments and beam shapes of the conventional multimode vertical cavity surface emitting lasers (VCSEL) is shown to be engineered in terms of their positions, widths, and polarizations by use of focused ion beam etching (FIBE). Several GaAs quantum well top-emitting devices with cavity diameters of 10 μm and 18 μm were investigated.

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Although there have been great advances in our understanding of the bacterial cytoskeleton, major gaps remain in our knowledge of its importance to virulence. In this study we have explored the contribution of the bacterial cytoskeleton to the ability of Salmonella to express and assemble virulence factors and cause disease. The bacterial actin-like protein MreB polymerises into helical filaments and interacts with other cytoskeletal elements including MreC to control cell-shape. As mreB appears to be an essential gene, we have constructed a viable ΔmreC depletion mutant in Salmonella. Using a broad range of independent biochemical, fluorescence and phenotypic screens we provide evidence that the Salmonella pathogenicity island-1 type three secretion system (SPI1-T3SS) and flagella systems are down-regulated in the absence of MreC. In contrast the SPI-2 T3SS appears to remain functional. The phenotypes have been further validated using a chemical genetic approach to disrupt the functionality of MreB. Although the fitness of ΔmreC is reduced in vivo, we observed that this defect does not completely abrogate the ability of Salmonella to cause disease systemically. By forcing on expression of flagella and SPI-1 T3SS in trans with the master regulators FlhDC and HilA, it is clear that the cytoskeleton is dispensable for the assembly of these structures but essential for their expression. As two-component systems are involved in sensing and adapting to environmental and cell surface signals, we have constructed and screened a panel of such mutants and identified the sensor kinase RcsC as a key phenotypic regulator in ΔmreC. Further genetic analysis revealed the importance of the Rcs two-component system in modulating the expression of these virulence factors. Collectively, these results suggest that expression of virulence genes might be directly coordinated with cytoskeletal integrity, and this regulation is mediated by the two-component system sensor kinase RcsC.

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One-cell-thick monolayers are the simplest tissues in multicellular organisms, yet they fulfill critical roles in development and normal physiology. In early development, embryonic morphogenesis results largely from monolayer rearrangement and deformation due to internally generated forces. Later, monolayers act as physical barriers separating the internal environment from the exterior and must withstand externally applied forces. Though resisting and generating mechanical forces is an essential part of monolayer function, simple experimental methods to characterize monolayer mechanical properties are lacking. Here, we describe a system for tensile testing of freely suspended cultured monolayers that enables the examination of their mechanical behavior at multi-, uni-, and subcellular scales. Using this system, we provide measurements of monolayer elasticity and show that this is two orders of magnitude larger than the elasticity of their isolated cellular components. Monolayers could withstand more than a doubling in length before failing through rupture of intercellular junctions. Measurement of stress at fracture enabled a first estimation of the average force needed to separate cells within truly mature monolayers, approximately ninefold larger than measured in pairs of isolated cells. As in single cells, monolayer mechanical properties were strongly dependent on the integrity of the actin cytoskeleton, myosin, and intercellular adhesions interfacing adjacent cells. High magnification imaging revealed that keratin filaments became progressively stretched during extension, suggesting they participate in monolayer mechanics. This multiscale study of monolayer response to deformation enabled by our device provides the first quantitative investigation of the link between monolayer biology and mechanics.

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Cyanobacteria perform photosynthesis and respiration in the thylakoid membrane, suggesting that the two processes are interlinked. However, the role of the respiratory electron transfer chain under natural environmental conditions has not been established. Through targeted gene disruption, mutants of Synechocystis sp. PCC 6803 were generated that lacked combinations of the three terminal oxidases: the thylakoid membrane-localized cytochrome c oxidase (COX) and quinol oxidase (Cyd) and the cytoplasmic membrane-localized alternative respiratory terminal oxidase. All strains demonstrated similar growth under continuous moderate or high light or 12-h moderate-light/dark square-wave cycles. However, under 12-h high-light/dark square-wave cycles, the COX/Cyd mutant displayed impaired growth and was completely photobleached after approximately 2 d. In contrast, use of sinusoidal light/dark cycles to simulate natural diurnal conditions resulted in little photobleaching, although growth was slower. Under high-light/dark square-wave cycles, the COX/Cyd mutant suffered a significant loss of photosynthetic efficiency during dark periods, a greater level of oxidative stress, and reduced glycogen degradation compared with the wild type. The mutant was susceptible to photoinhibition under pulsing but not constant light. These findings confirm a role for thylakoid-localized terminal oxidases in efficient dark respiration, reduction of oxidative stress, and accommodation of sudden light changes, demonstrating the strong selective pressure to maintain linked photosynthetic and respiratory electron chains within the thylakoid membrane. To our knowledge, this study is the first to report a phenotypic difference in growth between terminal oxidase mutants and wild-type cells and highlights the need to examine mutant phenotypes under a range of conditions.

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Jets from drop-on-demand inkjet print-heads consist of a main drop with a trailing filament, which either condenses into the main drop, or breaks up into satellite drops. Filament behaviour is quantitatively similar to that of larger, free symmetrical filamentscan be predicted from the aspect ratio and Ohnesorge number. Symmetrical filaments generated from inkjet print-heads show the same behaviour. A simple model, based on competition between the processes of axial shortening and radial necking, predicts the critical aspect ratio below which the jet condenses into a single drop. The success of this simple criterion supports the underlying physical model. © 2013 American Institute of Physics.

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In this communication, we report on the anisotropy of the superconducting properties of multifilamentary Bi-based tapes experimentally investigated by AC magnetic susceptibility measurements. The susceptibility $\chi= \chi' - j \chi''$ was measured using a commercial system and a couple of orthogonal pick-up coils. The $\chi''$ vs. temperature curves were shown to exhibit two peaks. The smaller of the peaks, occurring near T = 72K, was only visible for particular field directions and within a given frequency window. Such results point out the role played by the phase difference between the applied magnetic field and the internal magnetic field seen by the filaments.

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Several experimental techniques have been used in order to characterize the properties of multifilamentary Bi-2223 / Ag tapes. Pristine samples were investigated by electrical resistivity, current-voltage characteristics and DC magnetic moment measurements. Much emphasis is placed on comparing transport (direct) and magnetic (indirect) methods for determining the critical current density as well as the irreversibility line and resolving usual lacks of consistency due to the difference in measurement techniques and data analysis. The effect of an applied magnetic field, with various strengths and directions, is also studied and discussed. Next, the same combination of experiments was performed on bent tapes in order to bring out relevant information regarding the intergranular coupling. A modified Brandt model taking into account different types of defects within the superconducting filaments is proposed to reconciliate magnetic and transport data.