918 resultados para Laser scanning confocal microscope
Resumo:
The PITSLRE protein kinases are parts of the large family of p34cdc2-related kinases. During apoptosis induced by some stimuli, specific PITSLRE isoforms are cleaved by caspase to produce a protein that contains the C-terminal kinase domain of the PITSLRE proteins (p110C). The p110C induces apoptosis when it is ectopically expressed in Chinese hamster ovary cells. In our study, similar induction of this p110C was observed during anoikis in NIH3T3 cells. To investigate the molecular mechanism of apoptosis mediated by p110C, we used the yeast two-hybrid system to screen a human fetal liver cDNA library and identified p21-activated kinase 1 (PAK1) as an interacting partner of p110C. The association of p110C with PAK1 was further confirmed by in vitro binding assay, in vivo coimmunoprecipitation, and confocal microscope analysis. The interaction of p110C with PAK1 occurred within the residues 210-332 of PAK1. Neither association between p58PITSLRE or p110PITSLRE and PAK1 nor association between p110C and PAK2 or PAK3 was observed. Anoikis was increased and PAK1 activity was inhibited when NIH3T3 cells were transfected with p110C. Furthermore, the binding of p110C with PAK1 and inhibition of PAK1 activity were also observed during anoikis. Taken together, these data suggested that PAK1 might participate in the apoptotic pathway mediated by p110C.
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This paper reports the fabrication of SSOI (Silicon on Silicide On Insulator) substrates with active silicon regions only 0.5mum thick, incorporating LPCVD low resistivity tungsten silicide (WSix) as the buried layer. The substrates were produced using ion splitting and two stages of wafer bonding. Scanning acoustic microscope imaging confirmed that the bond interfaces are essentially void-free. These SSOI wafers are designed to be employed as substrates for mm-wave reflect-array diodes, and the required selective etch technology is described together with details of a suitable device.
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The process of making replicas of heritage has traditionally been developed by public agencies, corporations and museums and is not commonly used in schools. Currently there are technologies that allow creating cheap replicas. The new 3D reconstruction software, based on photographs and low cost 3D printers allow to make replicas at a cost much lower than traditional. This article describes the process of creating replicas of the sculpture Goslar Warrior of artist Henry Moore, located in Santa Cruz de Tenerife. To make this process, first, a digital model have been created using Autodesk Recap 360, Autodesk 123D Catch and Autodesk Meshmixer MarkerBot MakerWare applications. Physical replication, has been reproduced in polylactic acid (PLA) by MakerBot Replicator 2 3D printer. In addition, a cost analysis using, in one hand, the printer mentioned, and in the other hand, 3D printing services both online and local, is included. Finally, there has been a specific action with 141 students and 12 high school teachers, who filled a questionnary about the use of sculptural replicas in education.
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Purpose. Neovascularization occurs in response to tissue ischemia and growth factor stimulation. In ischemic retinopathies, however, new vessels fail to restore the hypoxic tissue; instead, they infiltrate the transparent vitreous. In a model of oxygen-induced retinopathy (OIR), TNFa and iNOS, upregulated in response to tissue ischemia, are cytotoxic and inhibit vascular repair. The aim of this study was to investigate the mechanism for this effect.
Methods. Wild-type C57/BL6 (WT) and TNFa-/- mice were subjected to OIR by exposure to 75% oxygen (postnatal days 7–12). The retinas were removed during the hypoxic phase of the model. Retinal cell death was determined by TUNEL staining, and the microglial cells were quantified after Z-series capture with a confocal microscope. In situ peroxynitrite and superoxide were measured by using the fluorescent dyes DCF and DHE. iNOS, nitrotyrosine, and arginase were analyzed by real-time PCR, Western blot analysis, and activity determined by radiolabeled arginine conversion. Astrocyte coverage was examined after GFAP immunostaining.
Results. The TNFa-/- animals displayed a significant reduction in TUNEL-positive apoptotic cells in the inner nuclear layer of the avascular retina compared with that in the WT control mice. The reduction coincided with enhanced astrocytic survival and an increase in microglial cells actively engaged in phagocytosing apoptotic debris that displayed low ROS, RNS, and NO production and high arginase activity.
Conclusions. Collectively, the results suggest that improved vascular recovery in the absence of TNFa is associated with enhanced astrocyte survival and that both phenomena are dependent on preservation of microglial cells that display an anti-inflammatory phenotype during the early ischemic phase of OIR.
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Polyamide and polystyrene particles were coated with titanium dioxide films by atomic layer deposition (ALD) and then melt-compounded to form polymer nanocomposites. The rheological properties of the ALD-created nanocomposite materials were characterized with a melt flow indexer, a melt flow spiral mould, and a rotational rheometer. The results suggest that the melt flow properties of polyamide nanocomposites were markedly better than those of pure polyamide and polystyrene nanocomposites. Such behavior was shown to originate in an uncontrollable decrease in the polyamide molecular weight, likely affected by a high thin-film impurity content, as shown in gel permeation chromatography (GPC) and scanning electron microscope (SEM) equipped with an energy-dispersive spectrometer. Transmission electron microscope image showed that a thin film grew on both studied polymer particles, and that subsequent melt-compounding was successful, producing well dispersed ribbon-like titanium dioxide with the titanium dioxide filler content ranging from 0.06 to 1.12wt%. Even though we used nanofillers with a high aspect ratio, they had only a minor effect on the tensile and flexural properties of the polystyrene nanocomposites. The mechanical behavior of polyamide nanocomposites was more complex because of the molecular weight degradation. Our approach here to form polymeric nanocomposites is one way to tailor ceramic nanofillers and form homogenous polymer nanocomposites with minimal work-related risks in handling powder form nanofillers. However, further research is needed to gauge the commercial potential of ALD-created nanocomposite materials. Copyright (C) 2011 John Wiley & Sons, Ltd.
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We simulate the localized surface plasmon resonances of an Au nanoparticle within tunnelling proximity of an Au substrate. The results demonstrate that the calculated resonance energies can be identified with those experimentally detected for light emission from the tip-sample junction of a scanning tunnelling microscope. Relative to the modes of an isolated nanoparticle these modes show significant red-shifting, extending further into the infrared with increasing radius, primarily due to a proximity-induced lowering of the effective bulk plasmon frequency. Spatial mapping of the field enhancement factor shows an oscillatory variation of the field, absent in the case of a dielectric substrate; also the degree of localization of the modes, and thus the resolution achievable electromagnetically, is shown to depend primarily on the nanoparticle radius, which is only weakly dependent on wavelength.
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In the present study we used a combination of patch clamping and fast confocal Ca2+ imaging to examine the effects of activators of the nitric oxide (NO)/cGMP pathway on pacemaker activity in freshly dispersed ICC from the rabbit urethra, using the amphotericin B perforated patch configuration of the patch-clamp technique. The nitric oxide donor, DEA-NO, the soluble guanylyl cyclase activator YC-1 and the membrane-permeant analogue of cGMP, 8-Br-cGMP inhibited spontaneous transient depolarizations (STDs) and spontaneous transient inward currents (STICs) recorded under current-clamp and voltage-clamp conditions, respectively. Caffeine-evoked Cl- currents were unaltered in the presence of SP-8-Br-PET-cGMPs, suggesting that activation of the cGMP/PKG pathway does not block Cl- channels directly or interfere with Ca2+ release via ryanodine receptors (RyR). However, noradrenaline-evoked Cl- currents were attenuated by SP-8-Br-PET-cGMPs, suggesting that activation of cGMP-dependent protein kinase (PKG) may modulate release of Ca2+ via IP3 receptors (IP3R). When urethral interstitial cells (ICC) were loaded with Fluo4-AM (2 microm), and viewed with a confocal microscope, they fired regular propagating Ca2+ waves, which originated in one or more regions of the cell. Application of DEA-NO or other activators of the cGMP/PKG pathway did not significantly affect the oscillation frequency of these cells, but did significantly reduce their spatial spread. These effects were mimicked by the IP3R blocker, 2-APB (100 microm). These data suggest that NO donors and activators of the cGMP pathway inhibit electrical activity of urethral ICC by reducing the spatial spread of Ca2+ waves, rather than decreasing wave frequency.
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We describe extensive studies on a family of perovskite oxides that are ferroelectric and ferromagnetic at ambient temperatures. The data include x-ray diffraction, Raman spectroscopy, measurements of ferroelectric and magnetic hysteresis, dielectric constants, Curie temperatures, electron microscopy
(both scanning electron microscope and transmission electron microscopy (TEM)) studies, and both longitudinal and transverse magnetoelectric constants a33 and a31. The study extends earlier work to lower Fe, Ta, and Nb concentrations at the B-site (from 15%–20% down to 5%). The magnetoelectric
constants increase supralinearly with Fe concentrations, supporting the earlier conclusions of a key role for Fe spin clustering. The room-temperature orthorhombic C2v point group symmetry inferred from earlier x-ray diffraction studies is confirmed via TEM, and the primitive unit cell size is found to be the basic perovskite Z¼1 structure of BaTiO3, also the sequence of phase transitions with increasing temperature from rhombohedral to orthorhombic to tetragonal to cubic mimics barium titanate.
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Purpose: The authors estimated the retinal nerve fiber layer height (RNFLH) measurements in patients with glaucoma compared with those in age-matched healthy subjects as obtained by the laser scanning tomography and assessed the relationship between RNFLH measurements and optic and visual field status. Methods: Parameters of optic nerve head topography and RNFLH were evaluated in 125 eyes of 21 healthy subjects and 104 patients with glaucoma using the Heidelberg Retina Tomograph ([HRT] Heidelberg Engineering GmbH, Heidelberg, Germany) for the entire disc area and for the superior 70°(50°temporal and 20°nasal to the vertical midline) and inferior 70°sectors of the optic disc. The mean deviation of the visual field, as determined by the Humphrey program 24-2 (Humphrey Instruments, Inc., San Leonardo, CA, U.S.A) was calculated in the entire field and in the superior and inferior Bjerrum area. Result: Retinal nerve fiber layer height parameters (mean RNFLH and RNFL cross-sectional area) were decreased significantly in patients with glaucoma compared with healthy individuals. Retinal nerve fiber layer height parameters was correlated strongly with rim volume, rim area, and cup/disc area ratio. Of the various topography measures, retinal nerve fiber layer (RNFL) parameters and cup/disc area ratio showed the strongest correlation with visual field mean deviation in patients with glaucoma. Conclusion: Retinal nerve fiber layer height measures were reduced substantially in patients with glaucoma compared with age-matched healthy subjects. Retinal nerve fiber layer height was correlated strongly with topographic optic disc parameters and visual field changes in patients with glaucoma.
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The structural, thermal, chemisorptive, and electronic properties of Ce on Pt{111} are studied by photoemission, Auger spectroscopy, scanning tunnel microscope (STM), and low-energy electron diffraction (LEED). Stranski-Krastanov-like growth of low-density Ce layers is accompanied by substantial valence charge transfer from Ce to Pt: in line with this, the measured dipole moment and polarizability of adsorbed Ce at low coverages are 7.2 x 10(-30) C m and similar to 1.3x10(-29) m(3), respectively. Pt-Ce intermixing commences at similar to 400 K and with increasing temperature a sequence of five different ordered surface alloys evolves. The symmetry, periodicities, and rotational epitaxy observed by LEED are in good accord with the STM data which reveal the true complexity of the system. The Various bimetallic surface phases are based on growth of crystalline Pt5Ce, a hexagonal layer structure consisting of alternating layers of Pt2Ce and Kagome nets of Pt atoms. This characteristic ABAB layered arrangement of the surface alloys is clearly imaged, and chemisorption data permit a distinction to be made between the more reactive Pt2Ce layer and the less reactive Pt Kagome net. Either type of layer can appear at the surface as the terminating structure, thicker films exhibiting unit mesh parameters characteristic of the bulk alloy.
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In this experimental study, diamond turning of single crystal 6H-SiC was performed at a cutting speed of 1 m/s on an ultra-precision diamond turning machine (Moore Nanotech 350 UPL) to elucidate the microscopic origin of ductile-regime machining. Distilled water (pH value 7) was used as a preferred coolant during the course of machining in order to improve the tribological performance. A high magnification scanning electron microscope (SEM FIB- FEI Quanta 3D FEG) was used to examine the cutting tool before and after the machining. A surface finish of Ra=9.2 nm, better than any previously reported value on SiC was obtained. Also, tremendously high cutting resistance was offered by SiC resulting in the observation of significant wear marks on the cutting tool just after 1 km of cutting length. It was found out through a DXR Raman microscope that similar to other classical brittle materials (silicon, germanium, etc.) an occurrence of brittle-ductile transition is responsible for the ductile-regime machining of 6H-SiC. It has also been demonstrated that the structural phase transformations associated with the diamond turning of brittle materials which are normally considered as a prerequisite to ductile-regime machining, may not be observed during ductile-regime machining of polycrystalline materials.
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The commonly used British Standard constant head triaxial permeability (BS) test, for permeability testing of fine grained soils, is known to have a relatively long test duration. Consequently, a reduction in the required time for permeability test provides potential cost savings, to the construction industry (specifically, for use during Construction Quality Control (CQA) of landfill mineral liners). The purpose of this article is to investigate and evaluate alternative short duration testing methods for the measurement of the permeability of fine grained soils.
As part of the investigation the feasibility of an existing method of short duration permeability test, known as the Accelerated Permeability (AP) test was assessed and compared with permeability measured using British Standard method (BS) and Ramp Accelerated Permeability (RAP). Four different fine grained materials, of a variety of physical properties were compacted at various moisture contents to produced analogous samples for testing using three the three different methodologies. Fabric analysis was carried out on specimens derived from post-test samples using Mercury Intrusion Porosimetry (MIP) and Scanning Electron Microscope (SEM) to assess the effects of testing methodology on soil structure. Results showed that AP testing in general under predicts permeability values derived from the BS test due to large changes in structure of the soil caused by AP test methodology, which is also validated using MIP and SEM observations. RAP testing, in general provides an improvement to the AP test but still under-predicts permeability values. The potential savings in test duration are shown to be relatively minimal for both the AP and RAP tests.
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The burial of objects (human remains, explosives, weapons) below or behind concrete, brick, plaster or tiling may be associated with serious crime and are difficult locations to search. These are quite common forensic search scenarios but little has been published on them to-date. Most documented discoveries are accidental or from suspect/witness testimony. The problem in locating such hidden objects means a random or chance-based approach is not advisable. A preliminary strategy is presented here, based on previous studies, augmented by primary research where new technology or applications are required. This blend allows a rudimentary search workflow, from remote desktop study, to non-destructive investigation through to recommendations as to how the above may inform excavation, demonstrated here with a case study from a homicide investigation. Published case studies on the search for human remains demonstrate the problems encountered when trying to find and recover sealed-in and sealed over locations. Established methods include desktop study, photography, geophysics and search dogs:these are integrated with new technology (LiDAR and laser scanning; photographic rectification; close quarter aerial imagery; ground-penetrating radar on walls and gamma-ray/neutron activation radiography) to propose this possible search strategy.
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This paper is an extension to an idea coined during the 13th EUSPEN Conference (P6.23) named "surface defect machining" (SDM). The objective of this work was to demonstrate how a conventional CNC turret lathe can be used to obtain ultra high precision machined surface finish on hard steels without recourse to a sophisticated ultra precision machine tool. An AISI 4340 hard steel (69 HRC) workpiece was machined using a CBN cutting tool with and without SDM. Post-machining measurements by a Form Talysurf and a Scanning Electron Microscope (FEI Quanta 3D) revealed that SDM culminates to several key advantages (i) provides better quality of the machined surface integrity and offers (ii) lowering feed rate to 5μm/rev to obtain a machined surface roughness of 30 nm (optical quality).
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A prism coupling arrangement is used to excite surface plasmons at the surface of a thin silver aim and a photon scanning tunnelling microscope is used to detect the evanescent field above the silver surface. Excitation of the silver/ air mode of interest is performed at lambda(1) = 632 . 8 nm using a tightly focused beam, while the control of the tip is effected by exciting a counter-propagating surface plasmon field at a different wavelength. lambda(2) = 543 . 5 nm, using an unfocused beam covering a macroscopic area. Propagation of the red surface plasmon is evidenced by an exponential tail extending away from the launch site, but this feature is abruptly truncated if the surface plasmon encounters the edge of the silver film - there is no specularly reflected 'beam'. Importantly, the radiative decay of the surface mode at the film edge is observable only at larger tip-sample separations, emphasizing the importance of accessing the mesoscopic regime.