Developmental Expression of Kv Potassium Channels at the Axon Initial Segment of Cultured Hippocampal Neurons

Axonal outgrowth and the formation of the axon initial segment (AIS) are early events in the acquisition of neuronal polarity. The AIS is characterized by a high concentration of voltage-dependent sodium and potassium channels. However, the specific ion channel subunits present and their precise localization in this axonal subdomain vary both during development and among the types of neurons, probably determining their firing characteristics in response to stimulation. Here, we characterize the developmental expression of different subfamilies of voltage-gated potassium channels in the AISs of cultured mouse hippocampal neurons, including subunits Kv1.2, Kv2.2 and Kv7.2. In contrast to the early appearance of voltage-gated sodium channels and the Kv7.2 subunit at the AIS, Kv1.2 and Kv2.2 subunits were tethered at the AIS only after 10 days in vitro. Interestingly, we observed different patterns of Kv1.2 and Kv2.2 subunit expression, with each confined to distinct neuronal populations. The accumulation of Kv1.2 and Kv2.2 subunits at the AIS was dependent on ankyrin G tethering, it was not affected by disruption of the actin cytoskeleton and it was resistant to detergent extraction, as described previously for other AIS proteins. This distribution of potassium channels in the AIS further emphasizes the heterogeneity of this structure in different neuronal populations, as proposed previously, and suggests corresponding differences in action potential regulation.

Colocalization of α-actinin and Synaptopodin in the Pyramidal Cell Axon Initial Segment

The cisternal organelle that resides in the axon initial segment (AIS) of neocortical and hippocampal pyramidal cells is thought to be involved in regulating the Ca(2+) available to maintain AIS scaffolding proteins, thereby preserving normal AIS structure and function. Through immunocytochemistry and correlative light and electron microscopy, we show here that the actin-binding protein ?-actinin is present in the typical cistenal organelle of rodent pyramidal neurons as well as in a large structure in the AIS of a subpopulation of layer V pyramidal cells that we have called the "giant saccular organelle." Indeed, this localization of ?-actinin in the AIS is dependent on the integrity of the actin cytoskeleton. Moreover, in the cisternal organelle of cultured hippocampal neurons, ?-actinin colocalizes extensively with synaptopodin, a protein that interacts with both actin and ?-actinin, and they appear concomitantly during the development of these neurons. Together, these results indicate that ?-actinin and the actin cytoskeleton are important components of the cisternal organelle that are probably required to stabilize the AIS.

System Engineering Approach to Initial Design of LEO Remote Sensing Missions

The satellite remote sensing missions are essential for long-term research around the condition of the earth resources and environment. On the other hand, in recent years the application of microsatellites is of interest in many space programs for their less cost and response time. In microsatellite remote sensing missions there are tight interrelations between different requirements such as orbital altitude, revisit time, mission life and spatial resolution. Also, all of these requirements can affect the whole system level design characteristics. In this work, the remote sensing microsatellite sizing process is divided into three major design disciplines; a) orbit design, b) payload sizing and c) bus sizing. Finally, some specific design cases are investigated inside the design space for evaluating the effect of different design variables on the satellite total mass. Considering the results of the work, it is concluded that applying a systematic approach at the initial design phase of such projects provides a good insight to the not clearly seen interactions inside their highly extended design space

Replication Data Management,needs and solutions: an initial evaluation of conceptual approaches for integrating heterogeneous replication study data

Replication Data Management (RDM) aims at enabling the use of data collections from several iterations of an experiment. However, there are several major challenges to RDM from integrating data models and data from empirical study infrastructures that were not designed to cooperate, e.g., data model variation of local data sources. [Objective] In this paper we analyze RDM needs and evaluate conceptual RDM approaches to support replication researchers. [Method] We adapted the ATAM evaluation process to (a) analyze RDM use cases and needs of empirical replication study research groups and (b) compare three conceptual approaches to address these RDM needs: central data repositories with a fixed data model, heterogeneous local repositories, and an empirical ecosystem. [Results] While the central and local approaches have major issues that are hard to resolve in practice, the empirical ecosystem allows bridging current gaps in RDM from heterogeneous data sources. [Conclusions] The empirical ecosystem approach should be explored in diverse empirical environments.

Molecular dynamics modeling and simulation of void growth in two dimensions

The mechanisms of growth of a circular void by plastic deformation were studied by means of molecular dynamics in two dimensions (2D). While previous molecular dynamics (MD) simulations in three dimensions (3D) have been limited to small voids (up to ≈10 nm in radius), this strategy allows us to study the behavior of voids of up to 100 nm in radius. MD simulations showed that plastic deformation was triggered by the nucleation of dislocations at the atomic steps of the void surface in the whole range of void sizes studied. The yield stress, defined as stress necessary to nucleate stable dislocations, decreased with temperature, but the void growth rate was not very sensitive to this parameter. Simulations under uniaxial tension, uniaxial deformation and biaxial deformation showed that the void growth rate increased very rapidly with multiaxiality but it did not depend on the initial void radius. These results were compared with previous 3D MD and 2D dislocation dynamics simulations to establish a map of mechanisms and size effects for plastic void growth in crystalline solids.

Simulación de la rotura de barras de acero perlítico sometidas a tracción mediante un modelo de fisura cohesiva. Validación experimental

Las probetas cilíndricas fabricadas con materiales metálicos de elevada ductilidad, como el aluminio o el cobre, sometidas a tracción suelen presentar una rotura comúnmente denominada rotura en copa y cono, debido a su geometría. Este tipo de rotura se reproduce numéricamente con éxito mediante el modelo de Gurson-Tvergaard- Needleman, cuya formulación matemática se basa en el fenómeno físico de nucleación, crecimiento y coalescencia de microhuecos. A diferencia de dichos materiales, las barras de acero perlítico, material con una ductilidad apreciable, presentan un frente de rotura plano que no puede simularse correctamente con los modelos antes mencionados, apareciendo una región interior de daño que, en principio, también puede atribuirse a un fenómeno de nucleación y crecimiento de microhuecos, mientras que en el exterior aparece una zona cuya micrografía permite asociar su rotura a un mecanismo de clivaje. En trabajos anteriores los autores han presentado un elemento de intercara cohesivo dependiente de la triaxialidad de tensiones que, incorporado a un código de elementos finitos, permite reproducir de forma razonable el daño que se desarrolla en la región interior mencionada. En este trabajo se presentan los resultados de una campaña experimental que permite validar el modelo desarrollado. Para ello, se ensayan probetas de diferentes diámetros y se comparan los resultados con los obtenidos numéricamente, empleando tres bases extensométricas diferentes en cada uno de los diámetros. Los resultados numéricos se ajustan razonablemente bien a los obtenidos experimentalmente.The cylindrical specimens made of high-ductility metallic materials, such as aluminium and copper, usually fail showing a fracture surface commonly known as cup-cone fracture because of its shape. This type of fracture is successfully reproduced using the Gurson-Tvergaard-Needleman model, which is based on the physical process of nucleation, growth and coalescence of microvoids. Unlike these materials, pearlitic steel bars, which are considerably ductile, show a flat fracture surface that cannot be correctly reproduced with the aforementioned models. In this flat fracture surface, a dark region can be observed in the centre of the specimen, which is the result of a process of nucleation and growth of microvoids, while in the rest of the fracture surface a different region can be identified, which a micrographic study reveals to be the result of a process of cleavage. In previous works, the authors presented a triaxiality-dependent cohesive interface element that, implemented in a finite element code, can reproduce in a reasonably accurate manner the damage that takes place in the dark region mentioned before. The results of an experimental campaign designed to validate the model are presented in this paper. For it, different diameter specimens are tested and these results are compared to those obtained with the numerical models, using three different initial lengths for the strain. Numerical results agree reasonably well with those obtained experimentally.

Simulación de la rotura de barras de acero perlítico sometidas a tracción mediante un modelo de fisura cohesiva. Validación experimental

Las probetas cilíndricas fabricadas con materiales metálicos de elevada ductilidad, como el aluminio o el cobre, sometidas a tracción suelen presentar una rotura comúnmente denominada rotura en copa y cono, debido a su geometría. Este tipo de rotura se reproduce numéricamente con éxito mediante el modelo de Gurson-Tvergaard- Needleman, cuya formulación matemática se basa en el fenómeno físico de nucleación, crecimiento y coalescencia de microhuecos. A diferencia de dichos materiales, las barras de acero perlítico, material con una ductilidad apreciable, presentan un frente de rotura plano que no puede simularse correctamente con los modelos antes mencionados, apareciendo una región interior de daño que, en principio, también puede atribuirse a un fenómeno de nucleación y crecimiento de microhuecos, mientras que en el exterior aparece una zona cuya micrografía permite asociar su rotura a un mecanismo de clivaje. En trabajos anteriores los autores han presentado un elemento de intercara cohesivo dependiente de la triaxialidad de tensiones que, incorporado a un código de elementos finitos, permite reproducir de forma razonable el daño que se desarrolla en la región interior mencionada. En este trabajo se presentan los resultados de una campaña experimental que permite validar el modelo desarrollado. Para ello, se ensayan probetas de diferentes diámetros y se comparan los resultados con los obtenidos numéricamente, empleando tres bases extensométricas diferentes en cada uno de los diámetros. Los resultados numéricos se ajustan razonablemente bien a los obtenidos experimentalmente.The cylindrical specimens made of high-ductility metallic materials, such as aluminium and copper, usually fail showing a fracture surface commonly known as cup-cone fracture because of its shape. This type of fracture is successfully reproduced using the Gurson-Tvergaard-Needleman model, which is based on the physical process of nucleation, growth and coalescence of microvoids. Unlike these materials, pearlitic steel bars, which are considerably ductile, show a flat fracture surface that cannot be correctly reproduced with the aforementioned models. In this flat fracture surface, a dark region can be observed in the centre of the specimen, which is the result of a process of nucleation and growth of microvoids, while in the rest of the fracture surface a different region can be identified, which a micrographic study reveals to be the result of a process of cleavage. In previous works, the authors presented a triaxiality-dependent cohesive interface element that, implemented in a finite element code, can reproduce in a reasonably accurate manner the damage that takes place in the dark region mentioned before. The results of an experimental campaign designed to validate the model are presented in this paper. For it, different diameter specimens are tested and these results are compared to those obtained with the numerical models, using three different initial lengths for the strain. Numerical results agree reasonably well with those obtained experimentally.

Discussion of “Initial Pore Pressure from Vertical Surface Loads” by Jacobo Bielak (September, 1982)

The author presents a very interesting application of the ideas developed by Scott to determine the initial pore pressure in excess of the hydrostatic pore pressure in linear, elastic, homogeneous and isotropic soil-skeleton. Scott demonstrates that under vertical surface loads the problem is governed by Laplace's equation. Nevertheless the writers' think that it could be interesting to state clearly the conditions under which this analogy can be applied.

A novel scanning lens instrument for evaluating Fresnel lens performance: equipment development and initial results

A system dedicated to the optical transmittance characterization of Fresnel lenses has been developed at NREL, in collaboration with the UPM. The system quantifies the optical efficiency of the lens by generating a performance map. The shape of the focused spot may also be analyzed to understand change in the lens performance. The primary instrument components (lasers and CCD detector) have been characterized to confirm their capability for performing optical transmittance measurements. Measurements performed on SoG and PMMA lenses subject to a variety of indoor conditions (e.g., UV and damp heat) identified differences in the optical efficiency of the evaluated lenses, demonstrating the ability of the Scanning Lens Instrument (SLI) to distinguish between the aged lenses.

An initial study on atmospheric pressure ion transport by laser ionization and electrostatic fields.

Laser ionization of mixtures of gases at atmospheric pressure and the subsequent transport through electrostatic field is studied. A prototype is designed to perform the transport and detection of the ions. Relevance of the composition of the mixture of gases and ionization parameters is shown

Low initial aspect-ratio direct-drive target designs for shock- or self-ignition in the context of the laser Megajoule

Analysis of low initial aspect ratio direct-drive target designs is carried out by varying the implosion velocity and the fuel mass. Starting from two different spherical targets with a given 300?g-DT mass, optimization of laser pulse and drive power allows to obtain a set of target seeds referenced by their peak implosion velocities and initial aspect ratio (A = 3 and A = 5). Self-ignition is achieved with higher implosion velocity for A = 5-design than for A = 3-design. Then, rescaling is done to extend the set of designs to a huge amount of mass, peak kinetic energies and peak areal densities. Self-ignition kinetic energy threshold Ek is characterized by a dependance of Ek ? v? with ?-values which depart from self-ignition models. Nevertheless, self-ignition energy is seen lower for smaller initial aspect ratio. An analysis of Two-Plasmons Decay threshold and Rayleigh?Taylor instability e-folding is carried out and it is shown that two-plasmon decay threshold is always overpassed for all designs. The hydrodynamic stability analysis is performed by embedded models to deal with linear and non-linear regime. It is found that the A = 5-designs are always at the limit of disruption of the shell.

Initial design with L2 Monge-Kantorovich theory for the Monge–Ampère equation method of freeform optics

The Monge–Ampère (MA) equation arising in illumination design is highly nonlinear so that the convergence of the MA method is strongly determined by the initial design. We address the initial design of the MA method in this paper with the L2 Monge-Kantorovich (LMK) theory, and introduce an efficient approach for finding the optimal mapping of the LMK problem. Three examples, including the beam shaping of collimated beam and point light source, are given to illustrate the potential benefits of the LMK theory in the initial design. The results show the MA method converges more stably and faster with the application of the LMK theory in the initial design.

Ultrafast signals in protein folding and the polypeptide contracted state

To test the significance of ultrafast protein folding signals (≪1 msec), we studied cytochrome c (Cyt c) and two Cyt c fragments with major C-terminal segments deleted. The fragments remain unfolded under all conditions and so could be used to define the unfolded baselines for protein fluorescence and circular dichroism (CD) as a function of denaturant concentration. When diluted from high to low denaturant in kinetic folding experiments, the fragments readjust to their new baseline values in a “burst phase” within the mixing dead time. The fragment burst phase reflects a contraction of the polypeptide from a more extended unfolded condition at high denaturant to a more contracted unfolded condition in the poorer, low denaturant solvent. Holo Cyt c exhibits fluorescence and CD burst phase signals that are essentially identical to the fragment signals over the whole range of final denaturant concentrations, evidently reflecting the same solvent-dependent, relatively nonspecific contraction and not the formation of a specific folding intermediate. The significance of fast folding signals in Cyt c and other proteins is discussed in relation to the hypothesis of an initial rate-limiting search-nucleation-collapse step in protein folding [Sosnick, T. R., Mayne, L. & Englander, S. W. (1996) Proteins Struct. Funct. Genet. 24, 413–426].

Surgical trial in traumatic intracerebral haemorrhage (STITCH) : A randomised controlled trial of early surgery compared with Initial conservative treatment

Funding This project was funded by the National Institute for Health Research (NIHR) Health Technology Assessment programme.

Flickering fusion pores comparable with initial exocytotic pores occur in protein-free phospholipid bilayers

For the act of membrane fusion, there are two competing, mutually exclusive molecular models that differ in the structure of the initial pore, the pathway for ionic continuity between formerly separated volumes. Because biological “fusion pores” can be as small as ionic channels or gap junctions, one model posits a proteinaceous initial fusion pore. Because biological fusion pore conductance varies widely, another model proposes a lipidic initial pore. We have found pore opening and flickering during the fusion of protein-free phospholipid vesicles with planar phospholipid bilayers. Fusion pore formation appears to follow the coalescence of contacting monolayers to create a zone of hemifusion where continuity between the two adherent membranes is lipidic, but not aqueous. Hypotonic stress, causing tension in the vesicle membrane, promotes complete fusion. Pores closed soon after opening (flickering), and the distribution of fusion pore conductance appears similar to the distribution of initial fusion pores in biological fusion. Because small flickering pores can form in the absence of protein, the existence of small pores in biological fusion cannot be an argument in support of models based on proteinaceous pores. Rather, these results support the model of a lipidic fusion pore developing within a hemifused contact site.