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Sample records for actinic flux density

  1. Influence of clouds on the spectral actinic flux density in the lower troposphere (INSPECTRO): overview of the field campaigns

    NASA Astrophysics Data System (ADS)

    Thiel, S.; Ammannato, L.; Bais, A.; Bandy, B.; Blumthaler, M.; Bohn, B.; Engelsen, O.; Gobbi, G. P.; Gröbner, J.; Jäkel, E.; Junkermann, W.; Kazadzis, S.; Kift, R.; Kjeldstad, B.; Kouremeti, N.; Kylling, A.; Mayer, B.; Monks, P. S.; Reeves, C. E.; Schallhart, B.; Scheirer, R.; Schmidt, S.; Schmitt, R.; Schreder, J.; Silbernagl, R.; Topaloglou, C.; Thorseth, T. M.; Webb, A. R.; Wendisch, M.; Werle, P.

    2008-03-01

    Ultraviolet radiation is the key factor driving tropospheric photochemistry. It is strongly modulated by clouds and aerosols. A quantitative understanding of the radiation field and its effect on photochemistry is thus only possible with a detailed knowledge of the interaction between clouds and radiation. The overall objective of the project INSPECTRO was the characterization of the three-dimensional actinic radiation field under cloudy conditions. This was achieved during two measurement campaigns in Norfolk (East Anglia, UK) and Lower Bavaria (Germany) combining space-based, aircraft and ground-based measurements as well as simulations with the one-dimensional radiation transfer model UVSPEC and the three-dimensional radiation transfer model MYSTIC. During both campaigns the spectral actinic flux density was measured at several locations at ground level and in the air by up to four different aircraft. This allows the comparison of measured and simulated actinic radiation profiles. In addition satellite data were used to complete the information of the three dimensional input data set for the simulation. A three-dimensional simulation of actinic flux density data under cloudy sky conditions requires a realistic simulation of the cloud field to be used as an input for the 3-D radiation transfer model calculations. Two different approaches were applied, to derive high- and low-resolution data sets, with a grid resolution of about 100 m and 1 km, respectively. The results of the measured and simulated radiation profiles as well as the results of the ground based measurements are presented in terms of photolysis rate profiles for ozone and nitrogen dioxide. During both campaigns all spectroradiometer systems agreed within ±10% if mandatory corrections e.g. stray light correction were applied. Stability changes of the systems were below 5% over the 4 week campaign periods and negligible over a few days. The J(O1D) data of the single monochromator systems can be

  2. Influence of clouds on the spectral actinic flux density in the lower troposphere (INSPECTRO): overview of the field campaigns

    NASA Astrophysics Data System (ADS)

    Thiel, S.; Ammannato, L.; Bais, A.; Bandy, B.; Blumthaler, M.; Bohn, B.; Engelsen, O.; Gobbi, G. P.; Gröbner, J.; Jäkel, E.; Junkermann, W.; Kazadzis, S.; Kift, R.; Kjeldstad, B.; Kouremeti, N.; Kylling, A.; Mayer, B.; Monks, P. S.; Reeves, C. E.; Schallhart, B.; Scheirer, R.; Schmidt, S.; Schmitt, R.; Schreder, J.; Silbernagl, R.; Topaloglou, C.; Thorseth, T. M.; Webb, A. R.; Wendisch, M.; Werle, P.

    2007-09-01

    Ultraviolet radiation is the key factor driving tropospheric photochemistry. It is strongly modulated by clouds and aerosols. A quantitative understanding of the radiation field and its effect on photochemistry is thus only possible with a detailed knowledge of the interaction between clouds and radiation. The overall objective of the project INSPECTRO was the characterization of the three-dimensional actinic radiation field under cloudy conditions. This was achieved during two measurement campaigns in Norfolk (East Anglia, UK) and Lower Bavaria (Germany) combining space-based, aircraft and ground-based measurements as well as simulations with the one-dimensional radiation transfer model UVSPEC and the three-dimensional radiation transfer model MYSTIC. During both campaigns the spectral actinic flux density was measured at several locations at ground level and in the air by up to four different aircraft. This allows the comparison of measured and simulated actinic radiation profiles. In addition satellite data were used to complete the information of the three dimensional input data set for the simulation. A three-dimensional simulation of actinic flux density data under cloudy sky conditions requires a realistic simulation of the cloud field to be used as an input for the 3-D radiation transfer model calculations. Two different approaches were applied, to derive high- and low-resolution data sets, with a grid resolution of about 100 m and 1 km, respectively. The results of the measured and simulated radiation profiles as well as the results of the ground based measurements are presented in terms of photolysis rate profiles for ozone and nitrogen dioxide. During both campaigns all spectroradiometer systems agreed within ±10% if mandatory corrections e.g. stray light correction were applied. Stability changes of the systems were below 5% over the 4 week campaign periods and negligible over a few days. The J(O1D) data of the single monochromator systems can be

  3. Aerosol properties derived from spectral actinic flux measurements

    NASA Astrophysics Data System (ADS)

    Stark, H.; Schmidt, K. S.; Pilewskie, P.; Cozic, J.; Wollny, A. G.; Brock, C. A.; Baynard, T.; Lack, D.; Parrish, D. D.; Fehsenfeld, F. C.

    2008-12-01

    Measurement of aerosol properties is very important for understanding climate change. Aerosol optical properties influence solar radiation throughout the troposphere. According to the Working Group I report of the intergovernmental panel for climate change [IPCC, 2007], aerosols have a direct radiative forcing of - 0.5±0.4 W/m2 with a medium to low level of scientific understanding. This relatively large uncertainty indicates the need for more frequent and precise measurements of aerosol properties. We will show how actinic flux measurements can be used to derive important optical aerosol parameters such as aerosol optical thickness and depth, surface albedo, angstrom exponent, radiative forcing by clouds and aerosols, aerosol extinction, and others. The instrument used for this study is a combination of two spectroradiometers measuring actinic flux in the ultraviolet and visible radiation range from 280 to 690 nm with a resolution of 1 nm. Actinic flux is measured as the radiation incident on a spherical surface with sensitivity independent of direction. In contrast, irradiance is measured as the radiation incident on a plane surface, which depends on the cosine of the incident angle. Our goal is to assess the capabilities of using spectral actinic flux measurements to derive various aerosol properties. Here we will compare 1) actinic flux measurements to irradiance measurements from the spectral solar flux radiometer (SSFR), 2) derived aerosol size distributions with measurements from a white light optical particle counter (WLOPC) and ultra high sensitivity aerosol size spectrometer (UHSAS), and 3) derived aerosol optical extinction with measurements from a cavity ringdown aerosol extinction spectrometer (CRD-AES). These comparisons will utilize data from three recent field campaigns over New England and the Atlantic Ocean (ICARTT 2004), Texas and the Gulf of Mexico during (TexAQS/GoMACCS 2006), and Alaska and the Arctic Ocean (ARCPAC 2008) when the instruments

  4. Investigation of the 3-D actinic flux field in mountainous terrain

    NASA Astrophysics Data System (ADS)

    Wagner, J. E.; Angelini, F.; Blumthaler, M.; Fitzka, M.; Gobbi, G. P.; Kift, R.; Kreuter, A.; Rieder, H. E.; Simic, S.; Webb, A.; Weihs, P.

    2011-11-01

    During three field campaigns spectral actinic flux was measured from 290-500 nm under clear sky conditions in Alpine terrain and the associated O3- and NO2-photolysis frequencies were calculated and the measurement products were then compared with 1-D- and 3-D-model calculations. To do this 3-D-radiative transfer model was adapted for actinic flux calculations in mountainous terrain and the maps of the actinic flux field at the surface, calculated with the 3-D-radiative transfer model, are given. The differences between the 3-D- and 1-D-model results for selected days during the campaigns are shown, together with the ratios of the modeled actinic flux values to the measurements. In many cases the 1-D-model overestimates actinic flux by more than the measurement uncertainty of 10%. The results of using a 3-D-model generally show significantly lower values, and can underestimate the actinic flux by up to 30%. This case study attempts to quantify the impact of snow cover in combination with topography on spectral actinic flux. The impact of snow cover on the actinic flux was ~ 25% in narrow snow covered valleys, but for snow free areas there were no significant changes due snow cover in the surrounding area and it is found that the effect snow-cover at distances over 5 km from the point of interest was below 5%. Overall the 3-D-model can calculate actinic flux to the same accuracy as the 1-D-model for single points, but gives a much more realistic view of the surface actinic flux field in mountains as topography and obstruction of the horizon are taken into account.

  5. Investigation of the 3-D actinic flux field in mountainous terrain

    PubMed Central

    Wagner, J.E.; Angelini, F.; Blumthaler, M.; Fitzka, M.; Gobbi, G.P.; Kift, R.; Kreuter, A.; Rieder, H.E.; Simic, S.; Webb, A.; Weihs, P.

    2011-01-01

    During three field campaigns spectral actinic flux was measured from 290–500 nm under clear sky conditions in Alpine terrain and the associated O3- and NO2-photolysis frequencies were calculated and the measurement products were then compared with 1-D- and 3-D-model calculations. To do this 3-D-radiative transfer model was adapted for actinic flux calculations in mountainous terrain and the maps of the actinic flux field at the surface, calculated with the 3-D-radiative transfer model, are given. The differences between the 3-D- and 1-D-model results for selected days during the campaigns are shown, together with the ratios of the modeled actinic flux values to the measurements. In many cases the 1-D-model overestimates actinic flux by more than the measurement uncertainty of 10%. The results of using a 3-D-model generally show significantly lower values, and can underestimate the actinic flux by up to 30%. This case study attempts to quantify the impact of snow cover in combination with topography on spectral actinic flux. The impact of snow cover on the actinic flux was ~ 25% in narrow snow covered valleys, but for snow free areas there were no significant changes due snow cover in the surrounding area and it is found that the effect snow-cover at distances over 5 km from the point of interest was below 5%. Overall the 3-D-model can calculate actinic flux to the same accuracy as the 1-D-model for single points, but gives a much more realistic view of the surface actinic flux field in mountains as topography and obstruction of the horizon are taken into account. PMID:26412915

  6. A phenomenological density-scaling approach to lamellipodial actin dynamics†

    PubMed Central

    Lewalle, Alexandre; Fritzsche, Marco; Wilson, Kerry; Thorogate, Richard; Duke, Tom; Charras, Guillaume

    2014-01-01

    The integration of protein function studied in vitro in a dynamic system like the cell lamellipodium remains a significant challenge. One reason is the apparent contradictory effect that perturbations of some proteins can have on the overall lamellipodium dynamics, depending on exact conditions. Theoretical modelling offers one approach for understanding the balance between the mechanisms that drive and regulate actin network growth and decay. Most models use a ‘bottom-up’ approach, involving explicitly assembling biochemical components to simulate observable behaviour. Their correctness therefore relies on both the accurate characterization of all the components and the completeness of the relevant processes involved. To avoid potential pitfalls due to this uncertainty, we used an alternative ‘top-down’ approach, in which measurable features of lamellipodium behaviour, here observed in two different cell types (HL60 and B16-F1), directly inform the development of a simple phenomenological model of lamellipodium dynamics. We show that the kinetics of F-actin association and dissociation scales with the local F-actin density, with no explicit location dependence. This justifies the use of a simplified kinetic model of lamellipodium dynamics that yields predictions testable by pharmacological or genetic intervention. A length-scale parameter (the lamellipodium width) emerges from this analysis as an experimentally accessible probe of network regulatory processes. PMID:25485077

  7. Investigation of the three-dimensional actinic flux field in mountainous terrain

    NASA Astrophysics Data System (ADS)

    Wagner, J. E.; Angelini, F.; Blumthaler, M.; Fitzka, M.; Gobbi, J. P.; Kift, R.; Kreuter, A.; Rieder, H. E.; Webb, A.; Weihs, P.

    2010-09-01

    Spectrally resolved high quality actinic flux measurements between 290 nm and 500 nm have been performed in complex Alpine terrain under clear sky conditions. A three-dimensional Monte Carlo radiative transfer model was adapted for actinic flux calculations in mountainous terrain. This model is used to study the impact of topography and surface albedo on surface spectral actinic flux and no2- and o3-photolysis rates. This approach leads to surface maps of actinic flux and photolysis rates. The typical high spatial variability due to altitude, snow cover and shading affects is very well reproduced in the model. By running the model in three modes (realistic, without topography, with albedo zero) one gets a good estimation of the impact of topography and surface albedo.

  8. Actinic flux measurements and photolysis frequencies enhancements near clouds during DC3 and TORERO

    NASA Astrophysics Data System (ADS)

    Hall, S. R.; Ullmann, K.; Schmidt, S.; Kindel, B. C.; Hair, J. W.

    2012-12-01

    Spectrally resolved up and down-welling actinic flux was measured from aircraft during the Deep Convective Clouds & Chemistry Experiment (DC3) and Tropical Ocean Troposphere Exchange of reactive halogen species and oxygenated VOC (TORERO) field campaigns. The measurements were made on the NASA DC-8 and NSF/NCAR G-V aircraft with the Charged coupled device Actinic Flux Spectroradiometer (CAFS)and the HIAPER Airborne Radiation Package (HARP), respectively. Improvements in this instrumentation and the data analysis provide for fast, accurate measurements from the aircraft. Photolysis frequencies calculated from the actinic flux show significant enhancements above clouds. The upwelling signal is enhanced by the high reflectivity of the cloud below. The downwelling is also enhanced due to backscatter of reflected light from the cloud top. Under specific conditions, including high sun and highly reflective clouds, upwelling actinic radiation may exceed the downwelling even with clear skies above. These conditions may have occurred during TORERO and DC3 resulting in regions of highly active photochemistry.

  9. Neuronal Actin Dynamics, Spine Density and Neuronal Dendritic Complexity Are Regulated by CAP2.

    PubMed

    Kumar, Atul; Paeger, Lars; Kosmas, Kosmas; Kloppenburg, Peter; Noegel, Angelika A; Peche, Vivek S

    2016-01-01

    Actin remodeling is crucial for dendritic spine development, morphology and density. CAP2 is a regulator of actin dynamics through sequestering G-actin and severing F-actin. In a mouse model, ablation of CAP2 leads to cardiovascular defects and delayed wound healing. This report investigates the role of CAP2 in the brain using Cap2(gt/gt) mice. Dendritic complexity, the number and morphology of dendritic spines were altered in Cap2(gt/gt) with increased number of excitatory synapses. This was accompanied by increased F-actin content and F-actin accumulation in cultured Cap2(gt/gt) neurons. Moreover, reduced surface GluA1 was observed in mutant neurons under basal condition and after induction of chemical LTP. Additionally, we show an interaction between CAP2 and n-cofilin, presumably mediated through the C-terminal domain of CAP2 and dependent on cofilin Ser3 phosphorylation. In vivo, the consequences of this interaction were altered phosphorylated cofilin levels and formation of cofilin aggregates in the neurons. Thus, our studies identify a novel role of CAP2 in neuronal development and neuronal actin dynamics. PMID:27507934

  10. Neuronal Actin Dynamics, Spine Density and Neuronal Dendritic Complexity Are Regulated by CAP2

    PubMed Central

    Kumar, Atul; Paeger, Lars; Kosmas, Kosmas; Kloppenburg, Peter; Noegel, Angelika A.; Peche, Vivek S.

    2016-01-01

    Actin remodeling is crucial for dendritic spine development, morphology and density. CAP2 is a regulator of actin dynamics through sequestering G-actin and severing F-actin. In a mouse model, ablation of CAP2 leads to cardiovascular defects and delayed wound healing. This report investigates the role of CAP2 in the brain using Cap2gt/gt mice. Dendritic complexity, the number and morphology of dendritic spines were altered in Cap2gt/gt with increased number of excitatory synapses. This was accompanied by increased F-actin content and F-actin accumulation in cultured Cap2gt/gt neurons. Moreover, reduced surface GluA1 was observed in mutant neurons under basal condition and after induction of chemical LTP. Additionally, we show an interaction between CAP2 and n-cofilin, presumably mediated through the C-terminal domain of CAP2 and dependent on cofilin Ser3 phosphorylation. In vivo, the consequences of this interaction were altered phosphorylated cofilin levels and formation of cofilin aggregates in the neurons. Thus, our studies identify a novel role of CAP2 in neuronal development and neuronal actin dynamics. PMID:27507934

  11. Fast two-stream method for computing diurnal-mean actinic flux in vertically inhomogeneous atmospheres

    NASA Technical Reports Server (NTRS)

    Filyushkin, V. V.; Madronich, S.; Brasseur, G. P.; Petropavlovskikh, I. V.

    1994-01-01

    Based on a derivation of the two-stream daytime-mean equations of radiative flux transfer, a method for computing the daytime-mean actinic fluxes in the absorbing and scattering vertically inhomogeneous atmosphere is suggested. The method applies direct daytime integration of the particular solutions of the two-stream approximations or the source functions. It is valid for any duration of period of averaging. The merit of the method is that the multiple scattering computation is carried out only once for the whole averaging period. It can be implemented with a number of widely used two-stream approximations. The method agrees with the results obtained with 200-point multiple scattering calculations. The method was also tested in runs with a 1-km cloud layer with optical depth of 10, as well as with aerosol background. Comparison of the results obtained for a cloud subdivided into 20 layers with those obtained for a one-layer cloud with the same optical parameters showed that direct integration of particular solutions possesses an 'analytical' accuracy. In the case of the source function interpolation, the actinic fluxes calculated above the one-layer and 20-layer clouds agreed within 1%-1.5%, while below the cloud they may differ up to 5% (in the worst case). The ways of enhancing the accuracy (in a 'two-stream sense') and computational efficiency of the method are discussed.

  12. 47 CFR 25.208 - Power flux density limits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Power flux density limits. 25.208 Section 25... COMMUNICATIONS Technical Standards § 25.208 Power flux density limits. (a) In the band 3650-4200 MHz, the power flux density at the Earth's surface produced by emissions from a space station for all conditions...

  13. The influence of the spatial resolution of topographic input data on the accuracy of 3-D UV actinic flux and irradiance calculations

    NASA Astrophysics Data System (ADS)

    Weihs, P.; Wagner, J. E.; Schreier, S. F.; Rieder, H. E.; Angelini, F.; Blumthaler, M.; Fitzka, M.; Gobbi, G. P.; Kift, R.; Kreuter, A.; Simic, S.; Webb, A. R.

    2012-03-01

    The aim of this study is to investigate the influence of the spatial resolution of a digital elevation map (DEM) on the three-dimensional (3-D) radiative transfer performance for both spectral ultraviolet (UV) irradiance and actinic flux at 305 nm. Model simulations were performed for clear sky conditions for three case studies: the first and second one using three sites in the Innsbruck area and the third one using three sites at the Sonnblick observatory and surrounding area. It was found that the DEM resolution may change the altitude at some locations by up to 500 m, resulting in changes in the sky obscured by the horizon of up to 15%. The geographical distribution of UV irradiance and actinic flux shows that with larger pixel size, uncertainties in UV irradiance and actinic flux determination of up to 100% are possible. These large changes in incident irradiance and actinic flux with changing pixel size are strongly connected to shading effects. The effect of the DEM pixel size on irradiance and actinic flux was studied at the six locations, and it was found that significant increases in irradiance and actinic flux with increasing DEM pixel size occurred at one valley location at high solar zenith angles in the Innsbruck area as well as for one steep valley location in the Sonnblick area. This increase in irradiance and actinic flux with increasing DEM resolution is most likely to be connected to shading effects affecting the reflections from the surroundings.

  14. The influence of the spatial resolution of topographic input data on the accuracy of 3-D UV actinic flux and irradiance calculations

    NASA Astrophysics Data System (ADS)

    Weihs, P.; Wagner, J. E.; Schreier, S. F.; Rieder, H. E.; Angelini, F.; Blumthaler, M.; Fitzka, M.; Gobbi, G. P.; Kift, R.; Kreuter, A.; Simic, S.; Webb, A. R.

    2011-10-01

    The aim of this study was to investigate the influence of the spatial resolution of a digital elevation map (DEM) on the three-dimensional (3-D) radiative transfer performance for both spectral ultraviolet (UV) irradiance and actinic flux at 305 nm. Model simulations were performed for clear sky conditions for three case studies: the first and second one using three sites in the Innsbruck area and the third one using three sites at the Sonnblick Observatory and surrounding area. It was found that DEM resolution may change the altitude at some locations by up to 500 m, resulting in changes in the sky obscured by the horizon of up to 15%. The geographical distribution of UV irradiance and actinic flux shows that with larger pixel size, uncertainties in UV irradiance and actinic flux determination of up to 100% are possible. These large changes in incident irradiance and actinic flux with changing pixel size are strongly connected to shading effects. The effect of DEM pixel size on irradiance and actinic flux was studied at the six locations, and it was found that significant increases in irradiance and actinic flux with increasing DEM pixel size occurred at one valley location at high solar zenith angles in the Innsbruck area as well as for one steep valley location in the Sonnblick area. This increase in irradiance and actinic flux with increasing DEM resolution is most likely to be connected to shading effects affecting the reflections from the surroundings.

  15. Association between tensin 1 and p130Cas at focal adhesions links actin inward flux to cell migration

    PubMed Central

    Zhao, Zhihai; Tan, Song Hui; Machiyama, Hiroaki; Kawauchi, Keiko; Araki, Keigo; Hirata, Hiroaki; Sawada, Yasuhiro

    2016-01-01

    ABSTRACT Cell migration is a highly dynamic process that plays pivotal roles in both physiological and pathological processes. We have previously reported that p130Cas supports cell migration through the binding to Src as well as phosphorylation-dependent association with actin retrograde flow at focal adhesions. However, it remains elusive how phosphorylated Cas interacts with actin cytoskeletons. We observe that the actin-binding protein, tensin 1, co-localizes with Cas, but not with its phosphorylation-defective mutant, at focal adhesions in leading regions of migrating cells. While a truncation mutant of tensin 1 that lacks the phosphotyrosine-binding PTB and SH2 domains (tensin 1-SH2PTB) poorly co-localizes or co-immunoprecitates with Cas, bacterially expressed recombinant tensin 1-SH2PTB protein binds to Cas in vitro in a Cas phosphorylation-dependent manner. Furthermore, exogenous expression of tensin 1-SH2PTB, which is devoid of the actin-interacting motifs, interferes with the Cas-driven cell migration, slows down the inward flux of Cas molecules, and impedes the displacement of Cas molecules from focal adhesions. Taken together, our results show that tensin 1 links inwardly moving actin cytoskeletons to phosphorylated Cas at focal adhesions, thereby driving cell migration. PMID:27029899

  16. Subsynaptic AMPA Receptor Distribution Is Acutely Regulated by Actin-Driven Reorganization of the Postsynaptic Density

    PubMed Central

    Kerr, Justin M.; Blanpied, Thomas A.

    2012-01-01

    AMPA receptors (AMPARs) mediate synaptic transmission and plasticity during learning, development, and disease. Mechanisms determining subsynaptic receptor position are poorly understood but are key determinants of quantal size. We used a series of live-cell, high-resolution imaging approaches to measure protein organization within single postsynaptic densities in rat hippocampal neurons. By photobleaching receptors in synapse subdomains, we found that most AMPARs do not freely diffuse within the synapse, indicating they are embedded in a matrix that determines their subsynaptic position. However, time lapse analysis revealed that synaptic AMPARs are continuously repositioned in concert with plasticity of this scaffold matrix rather than simply by free diffusion. Using a fluorescence correlation analysis, we found that across the lateral extent of single PSDs, component proteins were differentially distributed, and this distribution was continually adjusted by actin treadmilling. The C-terminal PDZ ligand of GluA1 did not regulate its mobility or distribution in the synapse. However, glutamate receptor activation promoted subsynaptic mobility. Strikingly, subsynaptic immobility of both AMPARs and scaffold molecules remained essentially intact even after loss of actin filaments. We conclude that receptors are actively repositioned at the synapse by treadmilling of the actin cytoskeleton, an influence which is transmitted only indirectly to receptors via the pliable and surprisingly dynamic internal structure of the PSD. PMID:22238102

  17. Measurement of Flux Density of Cas A at Low Frequencies

    NASA Astrophysics Data System (ADS)

    Patil, Ajinkya; Fisher, R.

    2012-01-01

    Cas A is used as a flux calibrator throughout the radio spectrum. Therefore it is important to know the spectral and secular variations in its flux density. Earlier observations by Scott et. al. (1969) and Baars et. al. (1972) suggested a secular decrease in flux density of Cas A at a rate of about 1% per year at all frequencies. However later observations by Erickson & Perley (1975) and Read (1977) indicated anomalously high flux from Cas A at 38 MHz. Also, these observations suggested that the original idea of faster decay of the flux density rate at low frequencies may be in error or that something more complex than simple decay is affecting the flux density at low frequencies. The source changes at 38 MHz still remains a mystery. We intend to present the results of follow up observations made from 1995 to 1998 with a three element interferometer in Green Bank operating in frequency range 30 to 120 MHz. We will discuss the problems at such low frequencies due to large beamwidth and unstable ionosphere. We will also discuss the strategies we have used so far to to find the flux density of Cas A by calculating the ratio of flux density of Cas A to that of Cyg A, assuming flux density of Cyg A to be constant. Above mentioned work was performed in summer student program sponsored by National Radio Astronomy Observatory.

  18. 47 CFR 25.208 - Power flux density limits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... citations affecting § 25.208, see the List of CFR Sections Affected, which appears in the Finding Aids... 47 Telecommunication 2 2013-10-01 2013-10-01 false Power flux density limits. 25.208 Section 25... COMMUNICATIONS Technical Standards § 25.208 Power flux density limits. (a) In the band 3650-4200 MHz, the...

  19. 47 CFR 25.208 - Power flux density limits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... propagation conditions. Editorial Note: For Federal Register citations affecting § 25.208, see the List of CFR... 47 Telecommunication 2 2014-10-01 2014-10-01 false Power flux density limits. 25.208 Section 25... COMMUNICATIONS Technical Standards § 25.208 Power flux density limits. (a) In the band 3650-4200 MHz, the...

  20. 47 CFR 25.208 - Power flux density limits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... dBW/m2/MHz. Editorial Note: For Federal Register citations affecting § 25.208, see the List of CFR... 47 Telecommunication 2 2012-10-01 2012-10-01 false Power flux density limits. 25.208 Section 25... COMMUNICATIONS Technical Standards § 25.208 Power flux density limits. (a) In the band 3650-4200 MHz, the...

  1. 47 CFR 25.208 - Power flux density limits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... dBW/m2/MHz. Editorial Note: For Federal Register citations affecting § 25.208, see the List of CFR... 47 Telecommunication 2 2011-10-01 2011-10-01 false Power flux density limits. 25.208 Section 25... COMMUNICATIONS Technical Standards § 25.208 Power flux density limits. (a) In the band 3650-4200 MHz, the...

  2. Correlation between -ray flux density and redshift for Fermi blazars

    NASA Astrophysics Data System (ADS)

    Xiao, Hu-Bing; Pei, Zhi-Yuan; Xie, Hong-Jing; Hao, Jing-Meng; Yang, Jiang-He; Yuan, Yu-Hai; Liu, Yi; Fan, Jun-Hui

    2015-09-01

    Blazars are strong -ray emitters, the -ray emissions are likely strongly beamed, therefore, one should use the intrinsic (de-beamed) emissions to investigate its emission nature. In this work, we compiled a sample of Fermi blazars with available beaming Doppler factors, , to investigate the correlation between -ray flux density, , and redshift, . The analysis shows that there is no correlation between and for the observed -ray flux density, but there is a clear strong correlation between the intrinsic flux densities, and . We also discussed the relationship of -ray luminosity and short time scale for the observed data and the intrinsic data. Our analysis suggests that the intrinsic -ray flux density obeys the flux density and redshift relation, and the jet in -rays maybe a continuous case. The intrinsic luminosity and the short time scales obey the Elliot and Shapiro relation and Abramowicz and Nobili relation as well.

  3. Refractive Interstellar Scintillation for Flux Density Variations of Two Pulsars

    NASA Astrophysics Data System (ADS)

    Zhou, Ai-Zhi; Wu, Xin-Ji; Esamdin, A.

    2003-08-01

    The flux density structure functions of PSRs B0525+21 and B2111+46 are calculated with the refractive interstellar scintillation (RISS) theory. The theoretical curves are in good agreement with observations [Astrophys. J. 539 (2000) 300] (hereafter S2000). The spectra of the electron density fluctuations both are of Kolmogorov spectra. We suggest that the flux density variations observed for these two pulsars are attributed to refractive interstellar scintillation, not to intrinsic variability.

  4. AN ACCURATE FLUX DENSITY SCALE FROM 1 TO 50 GHz

    SciTech Connect

    Perley, R. A.; Butler, B. J. E-mail: BButler@nrao.edu

    2013-02-15

    We develop an absolute flux density scale for centimeter-wavelength astronomy by combining accurate flux density ratios determined by the Very Large Array between the planet Mars and a set of potential calibrators with the Rudy thermophysical emission model of Mars, adjusted to the absolute scale established by the Wilkinson Microwave Anisotropy Probe. The radio sources 3C123, 3C196, 3C286, and 3C295 are found to be varying at a level of less than {approx}5% per century at all frequencies between 1 and 50 GHz, and hence are suitable as flux density standards. We present polynomial expressions for their spectral flux densities, valid from 1 to 50 GHz, with absolute accuracy estimated at 1%-3% depending on frequency. Of the four sources, 3C286 is the most compact and has the flattest spectral index, making it the most suitable object on which to establish the spectral flux density scale. The sources 3C48, 3C138, 3C147, NGC 7027, NGC 6542, and MWC 349 show significant variability on various timescales. Polynomial coefficients for the spectral flux density are developed for 3C48, 3C138, and 3C147 for each of the 17 observation dates, spanning 1983-2012. The planets Venus, Uranus, and Neptune are included in our observations, and we derive their brightness temperatures over the same frequency range.

  5. Electronic Flux Density beyond the Born-Oppenheimer Approximation.

    PubMed

    Schild, Axel; Agostini, Federica; Gross, E K U

    2016-05-19

    In the Born-Oppenheimer approximation, the electronic wave function is typically real-valued and hence the electronic flux density (current density) seems to vanish. This is unfortunate for chemistry, because it precludes the possibility to monitor the electronic motion associated with the nuclear motion during chemical rearrangements from a Born-Oppenheimer simulation of the process. We study an electronic flux density obtained from a correction to the electronic wave function. This correction is derived via nuclear velocity perturbation theory applied in the framework of the exact factorization of electrons and nuclei. To compute the correction, only the ground state potential energy surface and the electronic wave function are needed. For a model system, we demonstrate that this electronic flux density approximates the true one very well, for coherent tunneling dynamics as well as for over-the-barrier scattering, and already for mass ratios between electrons and nuclei that are much larger than the true mass ratios. PMID:26878256

  6. Correlated flux densities from VLBI observations with the DSN

    NASA Technical Reports Server (NTRS)

    Coker, R. F.

    1992-01-01

    Correlated flux densities of extragalactic radio sources in the very long baseline interferometry (VLBI) astrometric catalog are required for the VLBI tracking of Galileo, Mars Observer, and future missions. A system to produce correlated and total flux density catalogs was developed to meet these requirements. A correlated flux density catalog of 274 sources, accurate to about 20 percent, was derived from more than 5000 DSN VLBI observations at 2.3 GHz (S-band) and 8.4 GHz (X-band) using 43 VLBI radio reference frame experiments during the period 1989-1992. Various consistency checks were carried out to ensure the accuracy of the correlated flux densities. All observations were made on the California-Spain and California-Australia DSN baselines using the Mark 3 wideband data acquisition system. A total flux density catalog, accurate to about 20 percent, with data on 150 sources, was also created. Together, these catalogs can be used to predict source strengths to assist in the scheduling of VLBI tracking passes. In addition, for those sources with sufficient observations, a rough estimate of source structure parameters can be made.

  7. Magnetic Flux Density in the Heliosphere through Several Solar Cycles

    NASA Astrophysics Data System (ADS)

    Erdős, G.; Balogh, A.

    2014-01-01

    We studied the magnetic flux density carried by solar wind to various locations in the heliosphere, covering a heliospheric distance range of 0.3-5.4 AU and a heliolatitudinal range from 80° south to 80° north. Distributions of the radial component of the magnetic field, BR , were determined over long intervals from the Helios, ACE, STEREO, and Ulysses missions, as well as from using the 1 AU OMNI data set. We show that at larger distances from the Sun, the fluctuations of the magnetic field around the average Parker field line distort the distribution of BR to such an extent that the determination of the unsigned, open solar magnetic flux density from the average lang|BR |rang is no longer justified. We analyze in detail two methods for reducing the effect of fluctuations. The two methods are tested using magnetic field and plasma velocity measurements in the OMNI database and in the Ulysses observations, normalized to 1 AU. It is shown that without such corrections for the fluctuations, the magnetic flux density measured by Ulysses around the aphelion phase of the orbit is significantly overestimated. However, the matching between the in-ecliptic magnetic flux density at 1 AU (OMNI data) and the off-ecliptic, more distant, normalized flux density by Ulysses is remarkably good if corrections are made for the fluctuations using either method. The main finding of the analysis is that the magnetic flux density in the heliosphere is fairly uniform, with no significant variations having been observed either in heliocentric distance or heliographic latitude.

  8. Magnetic flux density in the heliosphere through several solar cycles

    SciTech Connect

    Erdős, G.; Balogh, A.

    2014-01-20

    We studied the magnetic flux density carried by solar wind to various locations in the heliosphere, covering a heliospheric distance range of 0.3-5.4 AU and a heliolatitudinal range from 80° south to 80° north. Distributions of the radial component of the magnetic field, B{sub R} , were determined over long intervals from the Helios, ACE, STEREO, and Ulysses missions, as well as from using the 1 AU OMNI data set. We show that at larger distances from the Sun, the fluctuations of the magnetic field around the average Parker field line distort the distribution of B{sub R} to such an extent that the determination of the unsigned, open solar magnetic flux density from the average (|B{sub R} |) is no longer justified. We analyze in detail two methods for reducing the effect of fluctuations. The two methods are tested using magnetic field and plasma velocity measurements in the OMNI database and in the Ulysses observations, normalized to 1 AU. It is shown that without such corrections for the fluctuations, the magnetic flux density measured by Ulysses around the aphelion phase of the orbit is significantly overestimated. However, the matching between the in-ecliptic magnetic flux density at 1 AU (OMNI data) and the off-ecliptic, more distant, normalized flux density by Ulysses is remarkably good if corrections are made for the fluctuations using either method. The main finding of the analysis is that the magnetic flux density in the heliosphere is fairly uniform, with no significant variations having been observed either in heliocentric distance or heliographic latitude.

  9. PHOTON FLUX DENSITY INFLUENCES GRASS RESPONSES TO EXTENDED PHOTOPERIOD

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant sensitivity to extended photoperiod has been well documented, with little attention to the possibility that quantum flux density used to extend photoperiod has an influence on the expression of photoperiod response. This study was undertaken with 4 grass species under field conditions to exami...

  10. Internal wave pressure, velocity, and energy flux from density perturbations

    NASA Astrophysics Data System (ADS)

    Allshouse, Michael R.; Lee, Frank M.; Morrison, Philip J.; Swinney, Harry L.

    2016-05-01

    Determination of energy transport is crucial for understanding the energy budget and fluid circulation in density varying fluids such as the ocean and the atmosphere. However, it is rarely possible to determine the energy flux field J =p u , which requires simultaneous measurements of the pressure and velocity perturbation fields p and u , respectively. We present a method for obtaining the instantaneous J (x ,z ,t ) from density perturbations alone: A Green's function-based calculation yields p ; u is obtained by integrating the continuity equation and the incompressibility condition. We validate our method with results from Navier-Stokes simulations: The Green's function method is applied to the density perturbation field from the simulations and the result for J is found to agree typically to within 1% with J computed directly using p and u from the Navier-Stokes simulation. We also apply the Green's function method to density perturbation data from laboratory schlieren measurements of internal waves in a stratified fluid and the result for J agrees to within 6 % with results from Navier-Stokes simulations. Our method for determining the instantaneous velocity, pressure, and energy flux fields applies to any system described by a linear approximation of the density perturbation field, e.g., to small-amplitude lee waves and propagating vertical modes. The method can be applied using our matlab graphical user interface EnergyFlux.

  11. Critical current density and flux pinning in an unconventional superconductor

    NASA Astrophysics Data System (ADS)

    Kumar, S.; Kaul, S. N.; Rodríguez Fernández, J.; Fernández Barquín, L.

    2009-12-01

    The functional dependence of the critical current density on magnetic field, J(H), observed at fixed temperatures in the unconventional type-II superconductor, LaAgMn ( c=0.1,0.2,0.3) alloys, but not the relative magnitude of J in different alloy compositions at any given temperature and field, is adequately described by the exponential-decay critical state model. In accordance with the predictions of the Kramer's flux-pinning model, the peak value of the pinning force density FPmax∝( with the exponent 1.7⩽m⩽2.8 and F/FPmax scales with h=H/Hc_2, where Hc_2 is the upper critical field. Irrespective of sample composition and temperature in the superconducting state, the pinning of the flux line lattice (FLL) dominates over the plastic FLL shear.

  12. Flux density calibration in diffuse optical tomographic systems

    NASA Astrophysics Data System (ADS)

    Biswas, Samir Kumar; Rajan, Kanhirodan; Vasu, Ram M.

    2013-02-01

    The solution of the forward equation that models the transport of light through a highly scattering tissue material in diffuse optical tomography (DOT) using the finite element method gives flux density (Φ) at the nodal points of the mesh. The experimentally measured flux (U) on the boundary over a finite surface area in a DOT system has to be corrected to account for the system transfer functions (R) of various building blocks of the measurement system. We present two methods to compensate for the perturbations caused by R and estimate true flux density (Φ) from Umeasuredcal. In the first approach, the measurement data with a homogeneous phantom (Umeasuredhomo) is used to calibrate the measurement system. The second scheme estimates the homogeneous phantom measurement using only the measurement from a heterogeneous phantom, thereby eliminating the necessity of a homogeneous phantom. This is done by statistically averaging the data (Umeasuredhetero) and redistributing it to the corresponding detector positions. The experiments carried out on tissue mimicking phantom with single and multiple inhomogeneities, human hand, and a pork tissue phantom demonstrate the robustness of the approach.

  13. Actinic Keratosis

    MedlinePlus

    ... rashes clinical tools newsletter | contact Share | Actinic Keratosis (Solar Keratosis) Information for adults A A A Actinic ... the touch. Overview Actinic keratoses, also known as solar keratoses, are small rough or scaly areas of ...

  14. Method for determining transport critical current densities and flux penetration depth in bulk superconductors

    NASA Technical Reports Server (NTRS)

    Israelsson, Ulf E. (Inventor); Strayer, Donald M. (Inventor)

    1992-01-01

    A contact-less method for determining transport critical current density and flux penetration depth in bulk superconductor material. A compressor having a hollow interior and a plunger for selectively reducing the free space area for distribution of the magnetic flux therein are formed of superconductor material. Analytical relationships, based upon the critical state model, Maxwell's equations and geometrical relationships define transport critical current density and flux penetration depth in terms of the initial trapped magnetic flux density and the ratio between initial and final magnetic flux densities whereby data may be reliably determined by means of the simple test apparatus for evaluating the current density and flux penetration depth.

  15. Spillage and flux density on a receiver aperture lip. [of solar thermal collector

    NASA Technical Reports Server (NTRS)

    Jaffe, L. D.

    1985-01-01

    In a dish-type point-focusing solar thermal collector, the spillage and the flux density on the receiver aperture lip are related in a very simple way, if the aperture is circular and centered on the optical axis. Specifically, the flux density on the lip is equal to the spillage times the peak flux density in the plane of the lip.

  16. Electromagnetic potentials basis for energy density and power flux

    NASA Astrophysics Data System (ADS)

    Puthoff, H. E.

    2016-09-01

    In rounding out the education of students in advanced courses in applied electromagnetics it is incumbent on us as mentors to raise issues that encourage appreciation of certain subtle aspects that are often overlooked during first exposure to the field. One of these has to do with the interplay between fields and potentials, with the latter often seen as just a convenient mathematical artifice useful in solving Maxwell’s equations. Nonetheless, to those practiced in application it is well understood that various alternatives in the use of fields and potentials are available within electromagnetic (EM) theory for the definitions of energy density, momentum transfer, EM stress–energy tensor, and so forth. Although the various options are all compatible with the basic equations of electrodynamics (e.g., Maxwell’s equations, Lorentz force law, gauge invariance), nonetheless certain alternative formulations lend themselves to being seen as preferable to others with regard to the transparency of their application to physical problems of interest. Here we argue for the transparency of an energy density/power flux option based on the EM potentials alone.

  17. Actinic keratosis

    MedlinePlus

    Solar keratosis; Sun-induced skin changes - keratosis; Keratosis - actinic (solar) ... Some actinic keratoses become squamous cell skin cancer . Have your health care provider look at all skin growths as soon as you find them. Your provider will ...

  18. Self-Organized Gels in DNA/F-Actin Mixtures without Crosslinkers: Networks of Induced Nematic Domains with Tunable Density

    NASA Astrophysics Data System (ADS)

    Lai, Ghee Hwee; Butler, John C.; Zribi, Olena V.; Smalyukh, Ivan I.; Angelini, Thomas E.; Purdy, Kirstin R.; Golestanian, Ramin; Wong, Gerard C. L.

    2008-11-01

    We examine mixtures of DNA and filamentous actin (F-actin) as a model system of like-charged rigid rods and flexible chains. Confocal microscopy reveals the formation of elongated nematic F-actin domains reticulated via defect-free vertices into a network embedded in a mesh of random DNA. Synchrotron x-ray scattering results indicate that the DNA mesh squeezes the F-actin domains into a nematic state with an interactin spacing that decreases with increasing DNA concentration as dactin∝ρDNA-1/2. Interestingly, the system changes from a counterion-controlled regime to a depletion-controlled regime with added salt, with drastic consequences for the osmotic pressure induced phase behavior.

  19. Calibration system for measuring the radon flux density.

    PubMed

    Onishchenko, A; Zhukovsky, M; Bastrikov, V

    2015-06-01

    The measurement of radon flux from soil surface is the useful tool for the assessment of radon-prone areas and monitoring of radon releases from uranium mining and milling residues. The accumulation chambers with hollow headspace and chambers with activated charcoal are the most used devices for these purposes. Systematic errors of the measurements strongly depend on the geometry of the chamber and diffusion coefficient of the radon in soil. The calibration system for the attestation of devices for radon flux measurements was constructed. The calibration measurements of accumulation chambers and chambers with activated charcoal were conducted. The good agreement between the results of 2D modelling of radon flux and measurements results was observed. It was demonstrated that reliable measurements of radon flux can be obtained by chambers with activated charcoal (equivalent volume ~75 l) or by accumulation chambers with hollow headspace of ~7-10 l and volume/surface ratio (height) of >15 cm. PMID:25977351

  20. Multispecies Density and Temperature Gradient Dependence of Quasilinear Particle and Energy Fluxes

    SciTech Connect

    G. Rewoldt; R.V. Budny; W.M. Tang

    2004-08-09

    The variations of the normalized quasilinear particle and energy fluxes with artificial changes in the density and temperature gradients, as well as the variations of the linear growth rates and real frequencies, for ion temperature gradient and trapped-electron modes, are calculated. The quasilinear fluxes are normalized to the total energy flux, summed over all species. Here, realistic cases for tokamaks and spherical torii are considered which have two impurity species. For situations where there are substantial changes in the normalized fluxes, the ''diffusive approximation,'' in which the normalized fluxes are taken to be linear in the gradients, is seen to be inaccurate. Even in the case of small artificial changes in density or temperature gradients, changes in the fluxes of different species (''off-diagonal'') generally are significant, or even dominant, compared to those for the same species (''diagonal'').

  1. Modeling of Fluctuating Mass Flux in Variable Density Flows

    NASA Technical Reports Server (NTRS)

    So, R. M. C.; Mongia, H. C.; Nikjooy, M.

    1983-01-01

    The approach solves for both Reynolds and Favre averaged quantities and calculates the scalar pdf. Turbulent models used to close the governing equations are formulated to account for complex mixing and variable density effects. In addition, turbulent mass diffusivities are not assumed to be in constant proportion to turbulent momentum diffusivities. The governing equations are solved by a combination of finite-difference technique and Monte-Carlo simulation. Some preliminary results on simple variable density shear flows are presented. The differences between these results and those obtained using conventional models are discussed.

  2. Gas Flux and Density Surrounding a Cylindrical Aperture in the Free Molecular Flow Regime

    NASA Technical Reports Server (NTRS)

    Soulas, George C.

    2011-01-01

    The equations for rigorously calculating the particle flux and density surrounding a cylindrical aperture in the free molecular flow regime are developed and presented. The fundamental equations for particle flux and density from a reservoir and a diffusely reflecting surface will initially be developed. Assumptions will include a Maxwell-Boltzmann speed distribution, equal particle and wall temperatures, and a linear flux distribution along the cylindrical aperture walls. With this information, the equations for axial flux and density surrounding a cylindrical aperture will be developed. The cylindrical aperture will be divided into multiple volumes and regions to rigorously determine the surrounding axial flux and density, and appropriate limits of integration will be determined. The results of these equations will then be evaluated. The linear wall flux distribution assumption will be assessed. The axial flux and density surrounding a cylindrical aperture with a thickness-to-radius ratio of 1.25 will be presented. Finally, the equations determined in this study will be verified using multiple methods.

  3. Rapid millimeter and centimeter band flux density increase in the gamma-ray blazar BL Lacertae

    NASA Astrophysics Data System (ADS)

    Hovatta, Talvikki; Richards, Joseph L.; Max-Moerbeck, Walter; Pearson, Timothy J.; Readhead, Anthony C. S.

    2012-11-01

    We have observed a rapid flux density increase of BL Lacertae (2200+420) at 15 GHz (2 cm) and 95 GHz (3 mm) following the report on highest millimeter flux density ever observed at the SMA (ATel #4557). Since 2009, BL Lacertae has been observed approximately twice per week at 15 GHz with the Owens Valley Radio Observatory (OVRO) 40m Telescope as part of our gamma-ray blazar monitoring program (Richards et al....

  4. Abnormal changes in the density of thermal neutron flux in biocenoses near the earth surface.

    PubMed

    Plotnikova, N V; Smirnov, A N; Kolesnikov, M V; Semenov, D S; Frolov, V A; Lapshin, V B; Syroeshkin, A V

    2007-04-01

    We revealed an increase in the density of thermal neutron flux in forest biocenoses, which was not associated with astrogeophysical events. The maximum spike of this parameter in the biocenosis reached 10,000 n/(sec x m2). Diurnal pattern of the density of thermal neutron flux depended only on the type of biocenosis. The effects of biomodulation of corpuscular radiation for balneology are discussed. PMID:18214289

  5. Extension of the absolute flux density scale to 22.285 GHz. [radio astronomy

    NASA Technical Reports Server (NTRS)

    Janssen, M. A.; Golden, L. M.; Welch, W. J.

    1974-01-01

    Extending the absolute flux density scale at microwave wavelengths, the absolute flux densities at 22.285 GHz of several standard sources were determined using the absolute calibrations of the 6.1 meter antenna of the Hat Creek Observatory. Interpolation formulas for each nonthermal standard source have been derived by combining these data with those determined at lower frequencies. The suitability of employing the standard sources for calibrating other antennas is discussed.

  6. Identification of the monitoring point density needed to reliably estimate contaminant mass fluxes

    NASA Astrophysics Data System (ADS)

    Liedl, R.; Liu, S.; Fraser, M.; Barker, J.

    2005-12-01

    Plume monitoring frequently relies on the evaluation of point-scale measurements of concentration at observation wells which are located at control planes or `fences' perpendicular to groundwater flow. Depth-specific concentration values are used to estimate the total mass flux of individual contaminants through the fence. Results of this approach, which is based on spatial interpolation, obviously depend on the density of the measurement points. Our contribution relates the accurracy of mass flux estimation to the point density and, in particular, allows to identify a minimum point density needed to achieve a specified accurracy. In order to establish this relationship, concentration data from fences installed in the coal tar creosote plume at the Borden site are used. These fences are characterized by a rather high density of about 7 points/m2 and it is reasonable to assume that the true mass flux is obtained with this point density. This mass flux is then compared with results for less dense grids down to about 0.1points/m2. Mass flux estimates obtained for this range of point densities are analyzed by the moving window method in order to reduce purely random fluctuations. For each position of the moving window the mass flux is estimated and the coefficient of variation (CV) is calculated to quantify variablity of the results. Thus, the CV provides a relative measure of accurracy in the estimated fluxes. By applying this approach to the Borden naphthalene plume at different times, it is found that the point density changes from sufficient to insufficient due to the temporally decreasing mass flux. By comparing the results of naphthalene and phenol at the same fence and at the same time, we can see that the same grid density might be sufficient for one compound but not for another. If a rather strict CV criterion of 5% is used, a grid of 7 points/m2 is shown to allow for reliable estimates of the true mass fluxes only in the beginning of plume development when

  7. Changes in magnetic flux density around fatigue crack tips of carbon tool steels

    NASA Astrophysics Data System (ADS)

    Honda, Takashi; Kida, Katsuyuki; Santos, Edson C.; Tanabe, Hirotaka

    2010-03-01

    Fatigue failure of steel occurs when small cracks form in a component and then continue to grow to a size large enough to cause failure. In order to understand the strength of steel components it is important to find the cracks which eventually grow to cause failures. However, at present, it is not easy to distinguish, in the early stages of growth, the cracks which will grow fast and cause failure. We hypothesized that it may be possible to distinguish them by comparing changes in the magnetic flux density around the tips of those cracks that grew large enough to cause failure. In order to measure these changes in magnetic flux density, we developed a scanning Hall probe microscope and observed the fatigue cracks growing from artificial slits in carbon tool steels (JIS SKS93). We also compared the changes in magnetic flux density around crack tips which grew under different loads and found that there is a strong correlation between the magnetic flux density, crack growth and stress intensity factors. In order to understand this relation, we measured the changes in the magnetic flux density and residual tensile stress by using an X-ray system, and found that the magnetic flux density changes not only in the plastic deformation area but also in the area of elastic stress field with increased stress.

  8. Changes in magnetic flux density around fatigue crack tips of carbon tool steels

    NASA Astrophysics Data System (ADS)

    Honda, Takashi; Kida, Katsuyuki; Santos, Edson C.; Tanabe, Hirotaka

    2009-12-01

    Fatigue failure of steel occurs when small cracks form in a component and then continue to grow to a size large enough to cause failure. In order to understand the strength of steel components it is important to find the cracks which eventually grow to cause failures. However, at present, it is not easy to distinguish, in the early stages of growth, the cracks which will grow fast and cause failure. We hypothesized that it may be possible to distinguish them by comparing changes in the magnetic flux density around the tips of those cracks that grew large enough to cause failure. In order to measure these changes in magnetic flux density, we developed a scanning Hall probe microscope and observed the fatigue cracks growing from artificial slits in carbon tool steels (JIS SKS93). We also compared the changes in magnetic flux density around crack tips which grew under different loads and found that there is a strong correlation between the magnetic flux density, crack growth and stress intensity factors. In order to understand this relation, we measured the changes in the magnetic flux density and residual tensile stress by using an X-ray system, and found that the magnetic flux density changes not only in the plastic deformation area but also in the area of elastic stress field with increased stress.

  9. Actinic keratosis

    MedlinePlus

    ... example, if you work outdoors) Had many severe sunburns early in life Are older Symptoms Actinic keratosis ... and tanning salons. Other things to know about sun exposure: Sun exposure is stronger in or near surfaces ...

  10. Actinic Cheilitis

    MedlinePlus

    ... is a precancerous condition related to cumulative lifetime sun exposure. The lower lip is most often affected. Individuals ... Wearing barrier clothing (eg, wide-brimmed hats) and sunscreen-containing lip balms can aid in preventing actinic ...

  11. 3D density imaging with muons flux measurements from underground galleries

    NASA Astrophysics Data System (ADS)

    Lesparre, Nolwenn; Cabrera, Justo; Marteau, Jacques

    2016-04-01

    Atmospheric muons flux measurements provide information on sub-surface density distribution, giving insights on the medium structure. We measured the muons flux from the underground galleries of the Tournemire experimental platform to image the medium between the galleries and the surface. The experiment aimed at evaluating the capacity of the method to detect the presence of discontinuities produced either by secondary strike-slip faults that present small vertical displacements or by a karstic network may be present at the level of an upper aquifer. Measurements were performed from three different sites so the trajectories of detected muons paths intersect in the medium. Such a configuration provided complementary information on the density distribution, offering the possibility to seek density variations at different depths. A specific calibration method was applied in order to interpolate the data acquired at different times with the same muons sensor. Muons flux measurements variations were then processed through a non-linear inversion, producing a 3D image of the density together with an evaluation of the different distinguished targets reliability. The density distribution showed the presence of a very low density region at the level of the upper aquifer, suggesting the presence of a karstic network hosting locally cavities. The trace of secondary strike-slip faults did not appear clearly on the image as the density contrast they produce might be too low compared to the signal to noise ratio present in the muons flux data. We propose different strategies to improve the density image accuracy.

  12. MAGNETIC FLUX DENSITY MEASURED IN FAST AND SLOW SOLAR WIND STREAMS

    SciTech Connect

    Erdos, G.; Balogh, A.

    2012-07-10

    The radial component of the heliospheric magnetic field vector is used to estimate the open magnetic flux density of the Sun. This parameter has been calculated using observations from the Ulysses mission that covered heliolatitudes from 80 Degree-Sign S to 80 Degree-Sign N, from 1990 to 2009 and distances from 1 to 5.4 AU, the Advanced Composition Explorer mission at 1 AU from 1997 to 2010, the OMNI interplanetary database from 1971, and the Helios 1 and 2 missions that covered the distance range from 0.3 to 1 AU. The flux density was found to be much affected by fluctuations in the magnetic field which make its calculated value dependent on heliospheric location, type of solar wind (fast or slow), and the level of solar activity. However, fluctuations are distributed symmetrically perpendicular to the average Parker direction. Therefore, distributions of the field vector in the two-dimensional plane defined by the radial and azimuthal directions in heliospheric coordinates provide a way to reduce the effects of the fluctuations on the measurement of the flux density. This leads to a better defined flux density parameter; the distributions modified by removing the effects of fluctuations then allow a clearer assessment of the dependence of the flux density on heliospheric location, solar wind type, and solar activity. This assessment indicates that the flux density normalized to 1 AU is independent of location and solar wind type (fast or slow). However, there is a residual dependence on solar activity which can be studied using the modified flux density measurements.

  13. Flux Density Variations in the Parkes Pulsar Timing Array Millisecond Pulsars

    NASA Astrophysics Data System (ADS)

    Spiewak, Renée; Shannon, Ryan; Hobbs, George; Kerr, Matthew

    2015-01-01

    Precise timing of an ensemble of pulsars spread across the sky (a pulsar timing array, PTA) can be used to search for gravitational waves. The Parkes Pulsar Timing Array project (PPTA) currently observes 23 pulsars with the Parkes Radio Telescope, largely in the southern sky, with the primary goal of searching for gravitational waves. The pulsars in the sample show large variations in flux density due to refractive scintillation in the interstellar medium (ISM). These flux variations cause timing uncertainty to vary by more than an order of magnitude. A better understanding of flux-density variations associated with the interstellar medium (ISM) is crucial for optimizing observing strategy and increase the sensitivity of the PPTA to gravitational waves. Flux-density variations can also potentially be caused by magnetospheric state changes. We use flux density time series and structure functions to examine both the properties of the ISM and search for intrinsic flux variation in these pulsars. We present intriguing features of the datasets and general implications of the results.

  14. Cluster electric current density measurements within a magnetic flux rope in the plasma sheet

    NASA Technical Reports Server (NTRS)

    Slavin, J. A.; Lepping, R. P.; Gjerloev, J.; Goldstein, M. L.; Fairfield, D. H.; Acuna, M. H.; Balogh, A.; Dunlop, M.; Kivelson, M. G.; Khurana, K.

    2003-01-01

    On August 22, 2001 all 4 Cluster spacecraft nearly simultaneously penetrated a magnetic flux rope in the tail. The flux rope encounter took place in the central plasma sheet, Beta(sub i) approx. 1-2, near the leading edge of a bursty bulk flow. The "time-of-flight" of the flux rope across the 4 spacecraft yielded V(sub x) approx. 700 km/s and a diameter of approx.1 R(sub e). The speed at which the flux rope moved over the spacecraft is in close agreement with the Cluster plasma measurements. The magnetic field profiles measured at each spacecraft were first modeled separately using the Lepping-Burlaga force-free flux rope model. The results indicated that the center of the flux rope passed northward (above) s/c 3, but southward (below) of s/c 1, 2 and 4. The peak electric currents along the central axis of the flux rope predicted by these single-s/c models were approx.15-19 nA/sq m. The 4-spacecraft Cluster magnetic field measurements provide a second means to determine the electric current density without any assumption regarding flux rope structure. The current profile determined using the curlometer technique was qualitatively similar to those determined by modeling the individual spacecraft magnetic field observations and yielded a peak current density of 17 nA/m2 near the central axis of the rope. However, the curlometer results also showed that the flux rope was not force-free with the component of the current density perpendicular to the magnetic field exceeding the parallel component over the forward half of the rope, perhaps due to the pressure gradients generated by the collision of the BBF with the inner magnetosphere. Hence, while the single-spacecraft models are very successful in fitting flux rope magnetic field and current variations, they do not provide a stringent test of the force-free condition.

  15. Bidirectional actin transport is influenced by microtubule and actin stability.

    PubMed

    Chetta, Joshua; Love, James M; Bober, Brian G; Shah, Sameer B

    2015-11-01

    Local and long-distance transport of cytoskeletal proteins is vital to neuronal maintenance and growth. Though recent progress has provided insight into the movement of microtubules and neurofilaments, mechanisms underlying the movement of actin remain elusive, in large part due to rapid transitions between its filament states and its diverse cellular localization and function. In this work, we integrated live imaging of rat sensory neurons, image processing, multiple regression analysis, and mathematical modeling to perform the first quantitative, high-resolution investigation of GFP-actin identity and movement in individual axons. Our data revealed that filamentous actin densities arise along the length of the axon and move short but significant distances bidirectionally, with a net anterograde bias. We directly tested the role of actin and microtubules in this movement. We also confirmed a role for actin densities in extension of axonal filopodia, and demonstrated intermittent correlation of actin and mitochondrial movement. Our results support a novel mechanism underlying slow component axonal transport, in which the stability of both microtubule and actin cytoskeletal components influence the mobility of filamentous actin. PMID:26043972

  16. Joint Measurements of Flare Flux Densities at 210 - 212 GHz by Two Different Radio Telescopes

    NASA Astrophysics Data System (ADS)

    Raulin, J.-P.; Trottet, G.; Giménez de Castro, G.; Lüthi, T.; Kaufmann, P.

    2014-04-01

    Multiple-beam observations of solar flares at submillimeter wavelengths need detection with at least four beams to derive the flux density of the emitting source, its size, and centroid position. When this condition is not fulfilled, the assumptions on the location and/or size of the emitting source have to be made in order to compute . Otherwise, only a flux density range can be estimated. We report on simultaneous flare observations at 212 and 210 GHz obtained by the Solar Submillimeter Telescope (SST) and the Bernese Multibeam Radiometer for Kosma (BEMRAK), respectively, during two solar events on 28 October 2003. For both events, BEMRAK utilized four beam information to calculate the source flux density F 210, its size and position. On the other hand, the SST observed the events with only one beam, at low solar elevation angles and during high atmospheric attenuation. Therefore, because of these poor observing conditions at 212 GHz, only a flux density range Δ F 212 could be estimated. The results show that Δ F 212 is within a factor of 2.5 of the flux density F 210. This factor can be significantly reduced ( e.g. 1.4 for one of the studied events) by an appropriate choice of the 212 GHz source position using flare observations at other wavelengths. By adopting the position and size of the 210 GHz source measured by BEMRAK, the flux density at 212 GHz, F 212b, is comparable to F 210 within the uncertainties, as expected. Therefore our findings indicate that even during poor observing conditions, the SST can provide an acceptable estimate of the flux density at 212 GHz. This is a remarkable fact since the SST and BEMRAK use quite different procedures for calibration and flux density determination. We also show that the necessary assumptions made on the size of the emitting source at 212 GHz in order to estimate its flux density are not critical, and therefore do not affect the conclusions of previous studies at this frequency.

  17. Actinic reticuloid

    SciTech Connect

    Marx, J.L.; Vale, M.; Dermer, P.; Ragaz, A.; Michaelides, P.; Gladstein, A.H.

    1982-09-01

    A 58-year-old man has his condition diagnosed as actinic reticuloid on the basis of clinical and histologic findings and phototesting data. He had clinical features resembling mycosis fungoides in light-exposed areas. Histologic findings disclosed a bandlike infiltrate with atypical mononuclear cells in the dermis and scattered atypical cells in the epidermis. Electron microscopy disclosed mononuclear cells with bizarre, convoluted nuclei, resembling cerebriform cells of Lutzner. Phototesting disclosed a diminished minimal erythemal threshold to UV-B and UV-A. Microscopic changes resembling actinic reticuloid were reproduced in this patient 24 and 72 hours after exposure to 15 minimal erythemal doses of UV-B.

  18. Concerning factors which determine whether flux-lattice shear or pin breaking limits the critical current density of superconductors

    SciTech Connect

    Welch, D.O.

    1992-10-01

    An elementary model is presented which illustrates the conditions under which flux-lattice shear, rather than pin breaking, limits the critical current density. An expression for the shear strength of the flux-lattice, based on the plasticity of metals and alloys, is used to derive the critical current density, including the effect of thermal activation in the flux creep regime.

  19. Concerning factors which determine whether flux-lattice shear or pin breaking limits the critical current density of superconductors

    SciTech Connect

    Welch, D.O.

    1992-01-01

    An elementary model is presented which illustrates the conditions under which flux-lattice shear, rather than pin breaking, limits the critical current density. An expression for the shear strength of the flux-lattice, based on the plasticity of metals and alloys, is used to derive the critical current density, including the effect of thermal activation in the flux creep regime.

  20. Effect of the Heat Flux Density on the Evaporation Rate of a Distilled Water Drop

    NASA Astrophysics Data System (ADS)

    Ponomarev, Konstantin; Orlova, Evgeniya; Feoktistov, Dmitry

    2016-02-01

    This paper presents the experimental dependence of the evaporation rate of a nondeaerated distilled water drop from the heat flux density on the surfaces of non-ferrous metals (copper and brass). A drop was placed on a heated substrate by electronic dosing device. To obtain drop profile we use a shadow optical system; drop symmetry was controlled by a high-speed video camera. It was found that the evaporation rate of a drop on a copper substrate is greater than on a brass. The evaporation rate increases intensively with raising volume of a drop. Calculated values of the heat flux density and the corresponding evaporation rates are presented in this work. The evaporation rate is found to increase intensively on the brass substrate with raising the heat flux density.

  1. Evaluation of soil heat flux density as a function of soil management practices

    NASA Astrophysics Data System (ADS)

    Moratiel Yugueros, R.; García Moreno, R.

    2012-04-01

    Soil energy is an important parameter in order to understand the flux of energy between the plant and the soil. This parameter could determine the potential for future production of soil. Pattern of surface energy flux varies depending on several factors, mainly on coverage. Also, this behaviour is strongly conditioned by the physical condition of soil. In order to evaluate the trend and behaviour of soil energy depending on soil coverage the aim of the present study was to evaluate soil heat flux density (G) in three different soil conditions depending on seasonal weather temperatures. Therefore, the authors monitored soil energy every half hour from soil located on bare soil, on soil covered by crops at root level and in between crop rows. The selected crop was corn. Soil heat flux density was measured with a heat flux plate sensor buried at a depth of 0.05 m in experimental sites. The change in heat storage in the soil layer above the heat flux plates was measured by inserting temperature sensors at an angle from near the bottom to near the top of the soil layer (above the plate sensor). The results indicated that the soil energy flux depends mainly on radiation and soil conditions. Although net radiation (Rn) was the same for all the sites, the evolution for G is different. Greater G fluctuation is produced in bared soils and decreases as soil is covered by the crops, especially at root level.

  2. DETECTION AND FLUX DENSITY MEASUREMENTS OF THE MILLISECOND PULSAR J2145–0750 BELOW 100 MHz

    SciTech Connect

    Dowell, J.; Taylor, G. B.; Craig, J.; Henning, P. A.; Schinzel, F.; Ray, P. S.; Blythe, J. N.; Clarke, T.; Helmboldt, J. F.; Ellingson, S. W.; Wolfe, C. N.; Lazio, T. J. W.; Stovall, K.

    2013-09-20

    We present flux density measurements and pulse profiles for the millisecond pulsar PSR J2145–0750 spanning 37 to 81 MHz using data obtained from the first station of the Long Wavelength Array. These measurements represent the lowest frequency detection of pulsed emission from a millisecond pulsar to date. We find that the pulse profile is similar to that observed at 102 MHz. We also find that the flux density spectrum between ≈40 MHz to 5 GHz is suggestive of a break and may be better fit by a model that includes spectral curvature with a rollover around 730 MHz rather than a single power law.

  3. Absolute Calibration of the Radio Astronomy Flux Density Scale at 22 to 43 GHz Using Planck

    NASA Astrophysics Data System (ADS)

    Partridge, B.; López-Caniego, M.; Perley, R. A.; Stevens, J.; Butler, B. J.; Rocha, G.; Walter, B.; Zacchei, A.

    2016-04-01

    The Planck mission detected thousands of extragalactic radio sources at frequencies from 28 to 857 GHz. Planck's calibration is absolute (in the sense that it is based on the satellite’s annual motion around the Sun and the temperature of the cosmic microwave background), and its beams are well characterized at sub-percent levels. Thus, Planck's flux density measurements of compact sources are absolute in the same sense. We have made coordinated Very Large Array (VLA) and Australia Telescope Compact Array (ATCA) observations of 65 strong, unresolved Planck sources in order to transfer Planck's calibration to ground-based instruments at 22, 28, and 43 GHz. The results are compared to microwave flux density scales currently based on planetary observations. Despite the scatter introduced by the variability of many of the sources, the flux density scales are determined to 1%–2% accuracy. At 28 GHz, the flux density scale used by the VLA runs 2%–3% ± 1.0% below Planck values with an uncertainty of +/- 1.0%; at 43 GHz, the discrepancy increases to 5%–6% ± 1.4% for both ATCA and the VLA.

  4. Dynamics of photosynthetic photon flux density (PPFD) and estimates in coastal northern California

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The seasonal trends and diurnal patterns of Photosynthetically Active Radiation (PAR) were investigated in the San Francisco Bay Area of Northern California from March through August in 2007 and 2008. During these periods, the daily values of PAR flux density (PFD), energy loading with PAR (PARE), a...

  5. Accurate Relations Between the Neutron Current Densities and the Neutron Fluxes

    SciTech Connect

    Ronen, Yigal

    2004-02-15

    Accurate relations between neutron current densities and neutron flux are obtained using the integral transport equation. Using these relations and Fick's Law, diffusion constants can be calculated. These diffusion constants are better than those usually used for the cases in which {sigma}{sub a}/{sigma}{sub s} is not small.

  6. Three-dimensional observations of magnetic flux density around fatigue crack tips of bearing steels

    NASA Astrophysics Data System (ADS)

    Kida, Katsuyuki; Santos, Edson C.; Honda, Takashi; Tanabe, Hirotaka

    2009-12-01

    Fatigue failure of steel occurs when small cracks form in a component and then continue to grow to a size large enough to cause failure. In order to understand the strength of steel components it is important to find these cracks. However, at present, it is not easy to distinguish the cracks that will grow fast and cause failure. We developed a three-dimensional scanning Hall probe microscope (3D-SHPM) and observed fatigue cracks at room temperature while they were growing. Four-point-bending fatigue tests were carried out using pre-cracked specimens (JIS-SUJ2, bearing steel). We observed the two-dimensional magnetic flux density distributions around the crack tips and found that there is a strong correlation between the changes in the magnetic flux densities and the crack growth. In order to understand this, we looked into all the three components of the magnetic flux densities, and found that they shape an arched bridge around a crack. We also found that the magnetic flux density moves in front of the crack tip along the crack growth direction.

  7. Three-dimensional observations of magnetic flux density around fatigue crack tips of bearing steels

    NASA Astrophysics Data System (ADS)

    Kida, Katsuyuki; Santos, Edson C.; Honda, Takashi; Tanabe, Hirotaka

    2010-03-01

    Fatigue failure of steel occurs when small cracks form in a component and then continue to grow to a size large enough to cause failure. In order to understand the strength of steel components it is important to find these cracks. However, at present, it is not easy to distinguish the cracks that will grow fast and cause failure. We developed a three-dimensional scanning Hall probe microscope (3D-SHPM) and observed fatigue cracks at room temperature while they were growing. Four-point-bending fatigue tests were carried out using pre-cracked specimens (JIS-SUJ2, bearing steel). We observed the two-dimensional magnetic flux density distributions around the crack tips and found that there is a strong correlation between the changes in the magnetic flux densities and the crack growth. In order to understand this, we looked into all the three components of the magnetic flux densities, and found that they shape an arched bridge around a crack. We also found that the magnetic flux density moves in front of the crack tip along the crack growth direction.

  8. Effect of Thermospheric Neutral Density upon Inner Trapped-belt Proton Flux

    NASA Technical Reports Server (NTRS)

    Wilson, Thomas L.; Lodhi, M. A. K.; Diaz, Abel B.

    2007-01-01

    We wish to point out that a secular change in the Earth's atmospheric neutral density alters charged-particle lifetime in the inner trapped radiation belts, in addition to the changes recently reported as produced by greenhouse gases. Heretofore, changes in neutral density have been of interest primarily because of their effect on the orbital drag of satellites. We extend this to include the orbital lifetime of charged particles in the lower radiation belts. It is known that the charged-belt population is coupled to the neutral density of the atmosphere through changes induced by solar activity, an effect produced by multiple scattering off neutral and ionized atoms along with ionization loss in the thermosphere where charged and neutral populations interact. It will be shown here that trapped-belt flux J is bivariant in energy E and thermospheric neutral density , as J(E,rho). One can conclude that proton lifetimes in these belts are also directly affected by secular changes in the neutral species populating the Earth s thermosphere. This result is a consequence of an intrinsic property of charged-particle flux, that flux is not merely a function of E but is dependent upon density rho when a background of neutrals is present.

  9. Long-term flux density variations of pulsars: Theoretical structure functions and comparisons with observations

    NASA Astrophysics Data System (ADS)

    Zhou, A. Z.; Wu, X. J.; Esamdin, A.

    2003-06-01

    By means of the refractive interstellar scintillation theory (RISS), the flux density structure functions of PSRs B1642-03, B0736-40, B0740-28 and B0329+54 are calculated and compared with the observations at 610 MHz by Stinebring et al. (\\cite{Stinebring00}, hereafter S2000). The theoretical results are in good agreement with observations and the spectra of the electron density fluctuation are all consistent with the Kolmogorov spectra. The theoretical modulation indices m are comparatively less sensitive to the distance H from the observer to the scattering screen but critically depend on the scattering strength line CN2. The structure function does not change remarkably with the variation of H if the scattering screen is closer to the pulsar than to the observer. The results in this paper indicate that the flux density variations observed for these four pulsars are due to a propagation effect (refractive scintillation), not to the intrinsic variability.

  10. Multiple-capillary measurement of RBC speed, flux, and density with optical coherence tomography

    PubMed Central

    Lee, Jonghwan; Wu, Weicheng; Lesage, Frederic; Boas, David A

    2013-01-01

    As capillaries exhibit heterogeneous and fluctuating dynamics even during baseline, a technique measuring red blood cell (RBC) speed and flux over many capillaries at the same time is needed. Here, we report that optical coherence tomography can capture individual RBC passage simultaneously over many capillaries located at different depths. Further, we demonstrate the ability to quantify RBC speed, flux, and linear density. This technique will provide a means to monitor microvascular flow dynamics over many capillaries at different depths at the same time. PMID:24022621

  11. Optimization of multiply acquired magnetic flux density Bz using ICNE-Multiecho train in MREIT

    NASA Astrophysics Data System (ADS)

    Nam, Hyun Soo; In Kwon, Oh

    2010-05-01

    The aim of magnetic resonance electrical impedance tomography (MREIT) is to visualize the electrical properties, conductivity or current density of an object by injection of current. Recently, the prolonged data acquisition time when using the injected current nonlinear encoding (ICNE) method has been advantageous for measurement of magnetic flux density data, Bz, for MREIT in the signal-to-noise ratio (SNR). However, the ICNE method results in undesirable side artifacts, such as blurring, chemical shift and phase artifacts, due to the long data acquisition under an inhomogeneous static field. In this paper, we apply the ICNE method to a gradient and spin echo (GRASE) multi-echo train pulse sequence in order to provide the multiple k-space lines during a single RF pulse period. We analyze the SNR of the measured multiple Bz data using the proposed ICNE-Multiecho MR pulse sequence. By determining a weighting factor for Bz data in each of the echoes, an optimized inversion formula for the magnetic flux density data is proposed for the ICNE-Multiecho MR sequence. Using the ICNE-Multiecho method, the quality of the measured magnetic flux density is considerably increased by the injection of a long current through the echo train length and by optimization of the voxel-by-voxel noise level of the Bz value. Agarose-gel phantom experiments have demonstrated fewer artifacts and a better SNR using the ICNE-Multiecho method. Experimenting with the brain of an anesthetized dog, we collected valuable echoes by taking into account the noise level of each of the echoes and determined Bz data by determining optimized weighting factors for the multiply acquired magnetic flux density data.

  12. The 3D heat flux density distribution on a novel parabolic trough wavy absorber

    NASA Astrophysics Data System (ADS)

    Demagh, Yassine; Kabar, Yassine; Bordja, Lyes; Noui, Samira

    2016-05-01

    The non-uniform concentrated solar flux distribution on the outer surface of the absorber pipe can lead to large circumferential gradient temperature and high concentrated temperature of the absorber pipe wall, which is one of the primary causes of parabolic trough solar receiver breakdown. In this study, a novel shape of the parabolic trough absorber pipe is proposed as a solution to well homogenize the solar flux distribution, as well as, the temperature in the absorber wall. The conventional straight absorber located along the focal line of the parabola is replaced by wavy one (invention patent by Y. Demagh [1]) for which the heat flux density distribution on the outer surface varies in both axial and azimuthal directions (3D) while it varies only in the azimuthal direction on the former (2D). As far as we know, there is not previous study which has used a longitudinally wavy pipe as an absorber into the parabolic trough collector unit.

  13. Simplified Solar Modulation Model of Inner Trapped Belt Proton Flux As a Function of Atmospheric Density

    NASA Technical Reports Server (NTRS)

    Wilson, Thomas L.; Lodhi, M. A. K.; Diaz, Abel B.

    2005-01-01

    No simple algorithm seems to exist for calculating proton fluxes and lifetimes in the Earth's inner, trapped radiation belt throughout the solar cycle. Most models of the inner trapped belt in use depend upon AP8 which only describes the radiation environment at solar maximum and solar minimum in Cycle 20. One exception is NOAAPRO which incorporates flight data from the TIROS/NOAA polar orbiting spacecraft. The present study discloses yet another, simple formulation for approximating proton fluxes at any time in a given solar cycle, in particular between solar maximum and solar minimum. It is derived from AP8 using a regression algorithm technique from nuclear physics. From flux and its time integral fluence, one can then approximate dose rate and its time integral dose. It has already been published in this journal that the absorbed dose rate, D, in the trapped belts exhibits a power law relationship, D = A(rho)(sup -n), where A is a constant, rho is the atmospheric density, and the index n is weakly dependent upon shielding. However, that method does not work for flux and fluence. Instead, we extend this idea by showing that the power law approximation for flux J is actually bivariant in energy E as well as density rho. The resulting relation is J(E,rho)approx.(sum of)A(E(sup n))rho(sup -n), with A itself a power law in E. This provides another method for calculating approximate proton flux and lifetime at any time in the solar cycle. These in turn can be used to predict the associated dose and dose rate.

  14. The brightness temperature of Venus and the absolute flux-density scale at 608 MHz.

    NASA Technical Reports Server (NTRS)

    Muhleman, D. O.; Berge, G. L.; Orton, G. S.

    1973-01-01

    The disk temperature of Venus was measured at 608 MHz near the inferior conjunction of 1972, and a value of 498 plus or minus 33 K was obtained using a nominal CKL flux-density scale. The result is consistent with earlier measurements, but has a much smaller uncertainty. Our theoretical model prediction is larger by a factor of 1.21 plus or minus 0.09. This discrepancy has been noticed previously for frequencies below 1400 MHz, but was generally disregarded because of the large observational uncertainties. No way could be found to change the model to produce agreement without causing a conflict with well-established properties of Venus. Thus it is suggested that the flux-density scale may require an upward revision, at least near this frequency, in excess of what has previously been considered likely.

  15. Double-cavity radiometer for high-flux density solar radiation measurements.

    PubMed

    Parretta, A; Antonini, A; Armani, M; Nenna, G; Flaminio, G; Pellegrino, M

    2007-04-20

    A radiometric method has been developed, suitable for both total power and flux density profile measurement of concentrated solar radiation. The high-flux density radiation is collected by a first optical cavity, integrated, and driven to a second optical cavity, where, attenuated, it is measured by a conventional radiometer operating under a stationary irradiation regime. The attenuation factor is regulated by properly selecting the aperture areas in the two cavities. The radiometer has been calibrated by a pulsed solar simulator at concentration levels of hundreds of suns. An optical model and a ray-tracing study have also been developed and validated, by which the potentialities of the radiometer have been largely explored. PMID:17415384

  16. Estimation of localized current anomalies in polymer electrolyte fuel cells from magnetic flux density measurements

    NASA Astrophysics Data System (ADS)

    Nara, Takaaki; Koike, Masanori; Ando, Shigeru; Gotoh, Yuji; Izumi, Masaaki

    2016-05-01

    In this paper, we propose novel inversion methods to estimate defects or localized current anomalies in membrane electrode assemblies (MEAs) in polymer electrolyte fuel cells (PEFCs). One method is an imaging approach with L1-norm regularization that is suitable for estimation of focal anomalies compared to Tikhonov regularization. The second is a complex analysis based method in which multiple pointwise current anomalies can be identified directly and algebraically from the measured magnetic flux density.

  17. A LOFAR census of non-recycled pulsars: average profiles, dispersion measures, flux densities, and spectra

    NASA Astrophysics Data System (ADS)

    Bilous, A. V.; Kondratiev, V. I.; Kramer, M.; Keane, E. F.; Hessels, J. W. T.; Stappers, B. W.; Malofeev, V. M.; Sobey, C.; Breton, R. P.; Cooper, S.; Falcke, H.; Karastergiou, A.; Michilli, D.; Osłowski, S.; Sanidas, S.; ter Veen, S.; van Leeuwen, J.; Verbiest, J. P. W.; Weltevrede, P.; Zarka, P.; Grießmeier, J.-M.; Serylak, M.; Bell, M. E.; Broderick, J. W.; Eislöffel, J.; Markoff, S.; Rowlinson, A.

    2016-06-01

    We present first results from a LOFAR census of non-recycled pulsars. The census includes almost all such pulsars known (194 sources) at declinations Dec > 8° and Galactic latitudes |Gb| > 3°, regardless of their expected flux densities and scattering times. Each pulsar was observed for ≥20 min in the contiguous frequency range of 110-188 MHz. Full-Stokes data were recorded. We present the dispersion measures, flux densities, and calibrated total intensity profiles for the 158 pulsars detected in the sample. The median uncertainty in census dispersion measures (1.5 × 10-3 pc cm-3) is ten times smaller, on average, than in the ATNF pulsar catalogue. We combined census flux densities with those in the literature and fitted the resulting broadband spectra with single or broken power-law functions. For 48 census pulsars such fits are being published for the first time. Typically, thechoice between single and broken power-laws, as well as the location of the spectral break, were highly influenced by the spectral coverage of the available flux density measurements. In particular, the inclusion of measurements below 100 MHz appears essential for investigating the low-frequency turnover in the spectra for most of the census pulsars. For several pulsars, we compared the spectral indices from different works and found the typical spread of values to be within 0.5-1.5, suggesting a prevailing underestimation of spectral index errors in the literature. The census observations yielded some unexpected individual source results, as we describe in the paper. Lastly, we will provide this unique sample of wide-band, low-frequency pulse profiles via the European Pulsar Network Database. Tables B.1-B.4 are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/591/A134

  18. Spatial and temporal variations in sap flux density in Japanese cedar (Cryptomeria japonica) trees, central Taiwan

    NASA Astrophysics Data System (ADS)

    Tseng, Han; Chiu, Chen-Wei; Wey, Tsong-Huei; Kume, Tomonori

    2013-04-01

    Sap flow measurement method is a technique widely used for measuring forest transpiration. However, variations in sap flow distribution can make accurately estimating individual tree-scale transpiration difficult. Significant spatial variations in sap flow across the sapwood within tree have been reported in many studies. In contrast, few studies have discussed azimuthal variations in sap flow, and even fewer have examined their seasonal change characteristics. This study was undertaken to clarify within-tree special and temporal variations in sap flow, and to propose an appropriate design for individual-tree scale transpiration estimates for Japanese cedar trees. The measurement was conducted in a Japanese cedar plantation located in Central Taiwan. Spatial distribution of sap flux density through the sapwood cross-section was measured using Granier's thermal dissipation technique. Sensors were installed at 1.3 m high on the east, west, north and south sides of the stem at 0-2 cm in 8 trees, and at 2-4 cm in the 6 larger trees. We found, in radial profile analysis, that sap flux densities measured at the depth of 2-4 cm were 50 % in average of those measured at depth of 0-2 cm. In azimuthal profile analysis, we found significant azimuthal variations in sap flux density. In one individual tree, the ratio of sap flux density on one aspect to another could be approximately 40-190 %, with no dependency on directions. Both radial and azimuthal profiles in most sample trees were fairly consistent throughout the measurement period. We concluded that radial and azimuthal variations in sap flow across sapwood might introduce significant errors in individual tree-scale transpiration estimations based on single point sap flow measurement, and seasonal change of within-tree spatial variations in sap flow could have insignificant impacts on accuracy of long-term individual tree-scale transpiration estimates. Keywords: transpiration, sap flow measurement, scaling up, sap flow

  19. The Importance of the Spatial Density of Satellite Measurements for the Retrieval of Spatial Flux Patterns

    NASA Astrophysics Data System (ADS)

    Marshall, J.; Nuñez Ramirez, T. G.; Kiemle, C.; Butz, A.; Hasekamp, O. P.; Ehret, G.; Heimann, M.

    2014-12-01

    Initial results from GOSAT flux inversions of column-integrated carbon dioxide suggest a significant redistribution of surface fluxes compared to inversions using only surface-based inversions as an observational constraint. New evidence suggests that this redistribution of fluxes is a robust feature, and is related to the increased spatial density of the measurements made available by remote sensing. However GOSAT's rather large measurement footprint and sparse sampling still provide poor coverage over many areas of the globe, particularly regions characterized by consistent cloud cover, such as the tropics, and all passive near-infrared sensors suffer from a seasonal sampling bias due to limited sunlight during high latitude winter. As such, errors in the pattern of retrieved fluxes may still be significant. Active sensors based on lidar do not suffer from the same seasonal (or diurnal) sampling biases, and their exceptionally small instantaneous field of view (~150 m) promises to greatly improve the spatial coverage of the measurements over partially cloudy regions. Using the case of MERLIN, a planned joint French-German lidar mission designed to measure XCH4, the implications of this increased spatial coverage is considered in an inverse modelling framework, and compared to presently available measurement coverage from the surface-based network and GOSAT. The gain in knowledge about the absolute size of the regional methane fluxes, particularly in currently undersampled regions such as the Arctic permafrost zones and tropical wetlands, is quantified.

  20. Uncertainty and Sensitivity of Alternative Rn-222 Flux Density Models Used in Performance Assessment

    SciTech Connect

    Greg J. Shott, Vefa Yucel, Lloyd Desotell Non-Nstec Authors: G. Pyles and Jon Carilli

    2007-06-01

    Performance assessments for the Area 5 Radioactive Waste Management Site on the Nevada Test Site have used three different mathematical models to estimate Rn-222 flux density. This study describes the performance, uncertainty, and sensitivity of the three models which include the U.S. Nuclear Regulatory Commission Regulatory Guide 3.64 analytical method and two numerical methods. The uncertainty of each model was determined by Monte Carlo simulation using Latin hypercube sampling. The global sensitivity was investigated using Morris one-at-time screening method, sample-based correlation and regression methods, the variance-based extended Fourier amplitude sensitivity test, and Sobol's sensitivity indices. The models were found to produce similar estimates of the mean and median flux density, but to have different uncertainties and sensitivities. When the Rn-222 effective diffusion coefficient was estimated using five different published predictive models, the radon flux density models were found to be most sensitive to the effective diffusion coefficient model selected, the emanation coefficient, and the radionuclide inventory. Using a site-specific measured effective diffusion coefficient significantly reduced the output uncertainty. When a site-specific effective-diffusion coefficient was used, the models were most sensitive to the emanation coefficient and the radionuclide inventory.

  1. Actinic Prurigo.

    PubMed

    Rodríguez-Carreón, Alma Angélica; Rodríguez-Lobato, Erika; Rodríguez-Gutiérrez, Georgina; Cuevas-González, Juan Carlos; Mancheno-Valencia, Alexandra; Solís-Arias, Martha Patricia; Vega-Memije, María Elisa; Hojyo-Tomoka, María Teresa; Domínguez-Soto, Luciano

    2015-01-01

    Actinic prurigo is an idiopathic photodermatosis that affects the skin, as well as the labial and conjunctival mucosa in indigenous and mestizo populations of Latin America. It starts predominantly in childhood, has a chronic course, and is exacerbated with solar exposure. Little is known of its pathophysiology, including the known mechanisms of the participation of HLA-DR4 and an abnormal immunologic response with increase of T CD4+ lymphocytes. The presence of IgE, eosinophils, and mast cells suggests that it is a hypersensitivity reaction (likely type IVa or b). The diagnosis is clinical, and the presence of lymphoid follicles in the mucosal histopathologic study of mucosa is pathognomonic. The best available treatment to date is thalidomide, despite its secondary effects. PMID:26861426

  2. [Actinic Keratosis].

    PubMed

    Dejaco, D; Hauser, U; Zelger, B; Riechelmann, H

    2015-07-01

    Actinic keratosis is a cutaneous lesion characterized by proliferation of atypical epidermal keratinocytes due to prolonged exposure to exogenous factors such as ultraviolet radiation. AKs are in-situ-squamous cell carcinomas (PEC) of the skin. AK typically presents as erythematous, scaly patch or papule (classic AK), occasionally as thick, adherent scale on an erythematous base. Mostly fair-skinned adults are affected. AKs typically occur in areas of frequent sun exposure (balding scalp, face, "H-region", lateral neck, décolleté, dorsum of the hand and lower extremities). Actinic Cheilitis is the term used for AKs appearing on the lips. The diagnosis of AK is based on clinical examination including inspection and palpation. The typical palpable rough surface of AK often precedes a visible lesion. Dermoscopy may provide additional information. If diagnosis is uncertain and invasion suspected, biopsy and histopathologic evaluation should be performed. The potential for progression to invasive PECs mandates therapeutic intervention. Treatment options include topical and systemic therapies. Topical therapies are classified into physical, medical and combined physical-chemical approaches and a sequential combination of treatment modalities is possible. Topical-physical cryotherapy is the treatment of choice for isolated, non-hypertrophic AK. Topical-medical treatment, e. g. 5-fluoruracil (5FU) cream or Imiquomod or Ingenolmebutat application is used for multiple, non-hypertrophic AKs. For hypertrophic AKs, a dehorning pretreatment with salicinated vaseline is recommended. Isolated hypertrophic AKs often need cryotherapy with prolonged freezing time or several consecutive applications. Sequentially combined approaches are recommended for multiple, hypertrophic AKs. Photodynamic therapy (PDT) as example for a combined physical-chemical approach is an established treatment for multiple, non-hypertrophic and hypertrophic AKs. Prevention includes avoidance of sun and

  3. Coupled-channels quantum theory of electronic flux density in electronically adiabatic processes: fundamentals.

    PubMed

    Diestler, D J

    2012-03-22

    The Born-Oppenheimer (BO) description of electronically adiabatic molecular processes predicts a vanishing electronic flux density (j(e)), =1/2∫dR[Δ(b) (x;R) - Δ(a) (x;R)] even though the electrons certainly move in response to the movement of the nuclei. This article, the first of a pair, proposes a quantum-mechanical "coupled-channels" (CC) theory that allows the approximate extraction of j(e) from the electronically adiabatic BO wave function . The CC theory is detailed for H(2)(+), in which case j(e) can be resolved into components associated with two channels α (=a,b), each of which corresponds to the "collision" of an "internal" atom α (proton a or b plus electron) with the other nucleus β (proton b or a). The dynamical role of the electron, which accommodates itself instantaneously to the motion of the nuclei, is submerged in effective electronic probability (population) densities, Δ(α), associated with each channel (α). The Δ(α) densities are determined by the (time-independent) BO electronic energy eigenfunction, which depends parametrically on the configuration of the nuclei, the motion of which is governed by the usual BO nuclear Schrödinger equation. Intuitively appealing formal expressions for the electronic flux density are derived for H(2)(+). PMID:22103768

  4. Models for the probability densities of the turbulent plasma flux in magnetized plasmas

    NASA Astrophysics Data System (ADS)

    Bergsaker, A. S.; Fredriksen, Å; Pécseli, H. L.; Trulsen, J. K.

    2015-10-01

    Observations of turbulent transport in magnetized plasmas indicate that plasma losses can be due to coherent structures or bursts of plasma rather than a classical random walk or diffusion process. A model for synthetic data based on coherent plasma flux events is proposed, where all basic properties can be obtained analytically in terms of a few control parameters. One basic parameter in the present case is the density of burst events in a long time-record, together with parameters in a model of the individual pulse shapes and the statistical distribution of these parameters. The model and its extensions give the probability density of the plasma flux. An interesting property of the model is a prediction of a near-parabolic relation between skewness and kurtosis of the statistical flux distribution for a wide range of parameters. The model is generalized by allowing for an additive random noise component. When this noise dominates the signal we can find a transition to standard results for Gaussian random noise. Applications of the model are illustrated by data from the toroidal Blaamann plasma.

  5. Enhanced magnetic flux density mapping using coherent steady state equilibrium signal in MREIT

    NASA Astrophysics Data System (ADS)

    Jeong, Woo Chul; Lee, Mun Bae; Sajib, Saurav Z. K.; Kim, Hyung Joong; Kwon, Oh In; Woo, Eung Je

    2016-03-01

    Measuring the z-component of magnetic flux density B = (Bx, By, Bz) induced by transversally injected current, magnetic resonance electrical impedance tomography (MREIT) aims to visualize electrical property (current density and/or conductivity distribution) in a three-dimensional imaging object. For practical implementations of MREIT technique, it is critical to reduce injection of current pulse within safety requirements. With the goal of minimizing the noise level in measured Bz data, we propose a new method to enhance the measure Bz data using steady-state coherent gradient multi-echo (SSC-GME) MR pulse sequence combining with injection current nonlinear encoding (ICNE) method in MREIT, where the ICNE technique injects current during a readout gradient to maximize the signal intensity of phase signal including Bz. The total phase offset in SSC-GME includes additional magnetic flux density due to the injected current, which is different from the phase signal for the conventional spoiled MR pulse sequence. We decompose the magnetization precession phase from the total phase offset including Bz and optimize Bz data using the steady-state equilibrium signal. Results from a real phantom experiment including different kinds of anomalies demonstrated that the proposed method enhanced Bz comparing to a conventional spoiled pulse sequence.

  6. Creation of a high density, high flux target plasmoid for magneto-inertial fusion

    NASA Astrophysics Data System (ADS)

    Weber, Thomas; Intrator, Thomas; Sears, Jason

    2011-10-01

    Magneto-inertial fusion utilizes embedded magnetic fields to reduce thermal transport and enhance alpha particle heating during an implosion reducing the required areal density, implosion speed, and convergence for fusion ignition. This enables the use of efficient inexpensive pulsed power, reducing the gain required for breakeven (e.g. ηG = 0 . 5 * 10 (MIF), = 0 . 05 * 100 (ICF)). The FRX-L and FRCHX experiments at Los Alamos National Laboratory and the Air Force Research Laboratory at Kirtland AFB are investigating a subset of MIF called Magnetized Target Fusion (MTF) in which a Field Reversed Configuration (FRC) plasmoid is injected into a converging solid, conductive liner and compressed to fusion conditions. Traditional FRC formation techniques utilizing ringing- θ pre-ionization have proved to be incapable of forming target plasmoids with enough density and magnetic flux, limiting the particle inventory, confinement, and lifetime. An alternative formation technique utilizing magnetoplasmadynamic arc sources has been developed to increase the density and trapped flux of the target plasmoid. Plasma source technology and operation are presented, as well as changes to the target formation process, plasmoid characteristics, and implications to MTF. Work supported by the DOE, OFES, under LANS Contract No. DE-AC52-06NA25369. Public release number LA-UR 11-03950.

  7. The Design of MACs (Minimal Actin Cortices)

    PubMed Central

    Vogel, Sven K; Heinemann, Fabian; Chwastek, Grzegorz; Schwille, Petra

    2013-01-01

    The actin cell cortex in eukaryotic cells is a key player in controlling and maintaining the shape of cells, and in driving major shape changes such as in cytokinesis. It is thereby constantly being remodeled. Cell shape changes require forces acting on membranes that are generated by the interplay of membrane coupled actin filaments and assemblies of myosin motors. Little is known about how their interaction regulates actin cell cortex remodeling and cell shape changes. Because of the vital importance of actin, myosin motors and the cell membrane, selective in vivo experiments and manipulations are often difficult to perform or not feasible. Thus, the intelligent design of minimal in vitro systems for actin-myosin-membrane interactions could pave a way for investigating actin cell cortex mechanics in a detailed and quantitative manner. Here, we present and discuss the design of several bottom-up in vitro systems accomplishing the coupling of actin filaments to artificial membranes, where key parameters such as actin densities and membrane properties can be varied in a controlled manner. Insights gained from these in vitro systems may help to uncover fundamental principles of how exactly actin-myosin-membrane interactions govern actin cortex remodeling and membrane properties for cell shape changes. © 2013 Wiley Periodicals, Inc. PMID:24039068

  8. A methodology for mapping forest latent heat flux densities using remote sensing

    NASA Technical Reports Server (NTRS)

    Pierce, Lars L.; Congalton, Russell G.

    1988-01-01

    Surface temperatures and reflectances of an upper elevation Sierran mixed conifer forest were monitored using the Thematic Mapper Simulator sensor during the summer of 1985 in order to explore the possibility of using remote sensing to determine the distribution of solar energy on forested watersheds. The results show that the method is capable of quantifying the relative energy allocation relationships between the two cover types defined in the study. It is noted that the method also has the potential to map forest latent heat flux densities.

  9. Effects of molecular symmetry on the directions of nuclear flux densities during tunnelling in double well potentials

    NASA Astrophysics Data System (ADS)

    Grohmann, Thomas; Manz, Jörn; Schild, Axel

    2013-08-01

    Coherent tunnelling in molecular systems with cyclic and non-cyclic symmetric double well potentials may proceed with similar nuclear densities, but with entirely different flux densities. For sufficiently high potential barriers, the nuclear densities may even become indistinguishable, whereas the patterns of the flux densities at a given time remain pincer-motion type for the cyclic systems, but unidirectional for the non-cyclic one. This effect is traced back to symmetry breaking of the cyclic to the non-cyclic model. Accordingly, nuclear flux densities are much more sensitive to symmetry breaking than nuclear densities. For a proof of principle, the phenomenon is demonstrated by means of three one-dimensional models. The cyclic model I represents torsion in oriented B2Cl2F2, the non-cyclic model II is constructed from I by symmetry breaking and the non-cyclic model III represents tunnelling by inversion of oriented NH3.

  10. The relationship between proton temperature and momentum flux density in the solar wind

    NASA Technical Reports Server (NTRS)

    Lopez, R. E.; Freeman, J. W.; Roelof, E. C.

    1986-01-01

    The relationship between proton temperature and momentum flux density/unit mass at 1 AU is examined using Helios 1 solar wind data from 1974 to 1980. In high-speed plasma (V greater than 500 km/s) T(p) increases with increasing n(0) V-squared, where n(0) and T(p) are the density and proton temperature at 1 AU and V is the flow speed. In lowspeed plasma (V less than 500 km/s), T(p) does not increase with increasing n(0) V-squared, and perhaps tends to decrease slightly. These basic relationships between T(p) and n(0) V-squared are not significantly affected by stream interactions. A qualitative explanation of these results is offered in the context of a solar wind model that includes deposition of momentum and energy extending well outward into the interplanetary medium.

  11. Thermally activated flux creep and critical current densities in high temperature superconductors

    NASA Astrophysics Data System (ADS)

    Matsushita, Teruo

    The effect of flux creep is discussed for projected strongly pinned oxide superconductors. It is determined, that if a superconducting wire with a critical current density higher than 10-billion A/sq m at 77 K and 5 T can be produced, the wire will be able to be applied to equipment at high fields; nonzero critical density will be obtained even at 77 K and high fields. The decay of persistent current is expected to be noticeable even in such strongly pinned superconductors, when those are used at 77 K. Although this will be managed in power equipment by lowering the operating current; variation in the magnetic field due to the variation in the current distribution inside superconducting wires appears to be unavoidable. It is suggested that an effort should be made to reduce the variation by reducing the diameter of the superconducting filaments.

  12. A turnkey data logger program for field-scale energy flux density measurements using eddy covariance and surface renewal

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Micrometeorological methods and ecosystem-scale energy and mass flux density measurements have become increasingly important in soil, agricultural, and environmental sciences. For many scientists without formal training in atmospheric science, these techniques are relatively inaccessible. Eddy cov...

  13. Arrangement Analysis of Leaves Optimized on Photon Flux Density or Photosynthetic Rate

    NASA Astrophysics Data System (ADS)

    Obara, Shin'ya; Tanno, Itaru

    By clarifying a plant evolutive process, useful information may be obtained on engineering. Consequently, an analysis algorithm that investigates the optimal arrangement of plant leaves was developed. In the developed algorithm, the Monte Carlo method is introduced and sunlight is simulated. Moreover, the arrangement optimization of leaves is analyzed using a Genetic Algorithm (GA). The number of light quanta (photon flux density) that reaches leaves, or the average photosynthetic rate of the same was set as the objective function, and leaf models of a dogwood and a ginkgo tree were analyzed. The number of leaf models was set between two to four, and the position of the leaf was expressed in terms of the angle of direction, elevation angle, rotation angle, and the representative length of the branch of a leaf. The chromosome model introduced into GA consists of information concerning the position of the leaf. Based on the analysis results, the characteristics of the leaf of an actual plant could be simulated by ensuring the algorithm had multiple constrained conditions. The optimal arrangement of leaves differs in maximization of the photon flux density, and that of the average value of a photosynthetic rate. Furthermore, the leaf form affecting the optimal arrangement of leave and also having a significant influence also on a photosynthetic rate was shown.

  14. Far scrape-off layer particle and heat fluxes in high density - High power scenarios

    NASA Astrophysics Data System (ADS)

    Müller, H. W.; Bernert, M.; Carralero, D.; Kallenbach, A.; Kurzan, B.; Scarabosio, A.; Sieglin, B.; Tophøj, L.; Vianello, N.; Wolfrum, E.

    2015-08-01

    The far scrape-off layer transport is studied in ASDEX Upgrade H-mode discharges with high divertor neutral density N0,div, high power across the separatrix Psep and nitrogen seeding to control the divertor temperature. Such conditions are expected for ITER but usually not investigated in terms of turbulent SOL transport. At high N0,div and Psep the H-mode discharges enter a regime of high cross-field particle and power transport in the SOL which is accompanied by a significant change of the turbulence characteristic analogous to the transition from conductive to convective transport in L-mode. Parallel particle and power flux densities of several 1023 m-2 s-1 and 10 MW m-2 have been detected about ∼40 to 45 mm outside the separatrix mapped to the outer mid-plane. The particle flux fall-off length reached up to 45 mm. This paper presents for the first time an empirical condition to enter the high transport regime in H-mode and the relation of this regime to changes in the filamentary transport.

  15. Microsystem for remote sensing of high energy radiation with associated extremely low photon flux densities

    NASA Astrophysics Data System (ADS)

    Otten, A.; Jain, V. K.

    2015-08-01

    This paper presents a microsystem for remote sensing of high energy radiation in extremely low flux density conditions. With wide deployment in mind, potential applications range from nuclear non-proliferation, to hospital radiation-safety. The daunting challenge is the low level of photon flux densities - emerging from a Scintillation Crystal (SC) on to a ~1 mm-square detector, which are a factor of 10000 or so lower than those acceptable to recently reported photonic chips (including `single-photon detection' chips), due to a combination of low Lux, small detector size, and short duration SC output pulses - on the order of 1 μs. These challenges are attempted to be overcome by the design of an innovative `System on a Chip' type microchip, with high detector sensitivity, and effective coupling from the SC to the photodetector. The microchip houses a tiny n+ diff p-epi photodiode (PD) as well as the associated analog amplification and other related circuitry, all fabricated in 0.5micron, 3-metal 2-poly CMOS technology. The amplification, together with pulse-shaping of the photocurrent-induced voltage signal, is achieved through a tandem of two capacitively coupled, double-cascode amplifiers. Included in the paper are theoretical estimates and experimental results.

  16. QUENCHING STAR FORMATION AT INTERMEDIATE REDSHIFTS: DOWNSIZING OF THE MASS FLUX DENSITY IN THE GREEN VALLEY

    SciTech Connect

    Goncalves, Thiago S.; Menendez-Delmestre, Karin; Martin, D. Christopher; Wyder, Ted K.; Koekemoer, Anton

    2012-11-01

    The bimodality in galaxy properties has been observed at low and high redshifts, with a clear distinction between star-forming galaxies in the blue cloud and passively evolving objects in the red sequence; the absence of galaxies with intermediate properties indicates that the quenching of star formation and subsequent transition between populations must happen rapidly. In this paper, we present a study of over 100 transiting galaxies in the so-called green valley at intermediate redshifts (z {approx} 0.8). By using very deep spectroscopy with the DEIMOS instrument at the Keck telescope we are able to infer the star formation histories of these objects and measure the stellar mass flux density transiting from the blue cloud to the red sequence when the universe was half its current age. Our results indicate that the process happened more rapidly and for more massive galaxies in the past, suggesting a top-down scenario in which the massive end of the red sequence is forming first. This represents another aspect of downsizing, with the mass flux density moving toward smaller galaxies in recent times.

  17. Density effect on critical current density and flux pinning properties of polycrystalline SmFeAsO1 - xFx superconductor

    NASA Astrophysics Data System (ADS)

    Ding, Y.; Sun, Y.; Zhuang, J. C.; Cui, L. J.; Shi, Z. X.; Sumption, M. D.; Majoros, M.; Susner, M. A.; Kovacs, C. J.; Li, G. Z.; Collings, E. W.; Ren, Z. A.

    2011-12-01

    A series of polycrystalline SmFeAs1 - xOx bulks was prepared to systematically investigate the influence of sample density on flux pinning properties. Different sample densities were achieved by controlling the pelletizing pressure. The superconducting volume fraction, the critical current densities Jcm and the flux pinning force densities Fp were estimated from the magnetization measurements. Experimental results show that: (1) the superconducting volume fraction increases with the increasing of sample density; (2) the Jcm values have a similar trend except for the sample with very high density due to different connectivity and pinning mechanisms, moreover, the Jcm(B) curve develops a peak effect at approximately the same field at which the high density sample shows a kink; (3) the Fp(B) curve of the high density sample shows a low-field peak and a high-field peak at several temperatures, which can be explained by improved intergranular current, while only one peak can be observed in Fp(B) of the low density samples. Based on the scaling behaviour of flux pinning force densities, the main intragranular pinning is normal point pinning.

  18. Photospheric Vertical Current Density and Overlying Atmospheric Activity in an Emerging Flux Region

    NASA Astrophysics Data System (ADS)

    Georgoulis, M. K.; Rust, D. M.; Bernasconi, P. N.; Schmieder, B.

    2002-05-01

    Using high-resolution vector magnetograms obtained by the balloon-borne Flare Genesis Experiment (FGE), we construct maps of the vertical current density in the emerging flux region NOAA 8844. The vertical current density has been decomposed into components that are field-aligned and perpendicular to the magnetic field, thus allowing a straightforward identification of force-free areas, as well as of areas where the force-free approximation breaks down. Small-scale chromospheric activity, such as H α Ellerman bombs and Ultraviolet bright points in 1600 Åshow a remarkable correlation with areas of strong current density. Simultaneous data of overlying coronal loops, observed by TRACE in the Extreme Ultraviolet (171 Åand 195 Å), have been carefully co-aligned with the FGE photospheric maps. We find that the footpoints of the TRACE loops always coincide with strong vertical currents and enhancements of the current helicity density. We also investigate whether the force-free approximation is valid on the photosphere during various evolutionary stages of the active region.

  19. Integration of linear and dendritic actin nucleation in Nck-induced actin comets

    PubMed Central

    Borinskaya, Sofya; Velle, Katrina B.; Campellone, Kenneth G.; Talman, Arthur; Alvarez, Diego; Agaisse, Hervé; Wu, Yi I.; Loew, Leslie M.; Mayer, Bruce J.

    2016-01-01

    The Nck adaptor protein recruits cytosolic effectors such as N-WASP that induce localized actin polymerization. Experimental aggregation of Nck SH3 domains at the membrane induces actin comet tails—dynamic, elongated filamentous actin structures similar to those that drive the movement of microbial pathogens such as vaccinia virus. Here we show that experimental manipulation of the balance between unbranched/branched nucleation altered the morphology and dynamics of Nck-induced actin comets. Inhibition of linear, formin-based nucleation with the small-molecule inhibitor SMIFH2 or overexpression of the formin FH1 domain resulted in formation of predominantly circular-shaped actin structures with low mobility (actin blobs). These results indicate that formin-based linear actin polymerization is critical for the formation and maintenance of Nck-dependent actin comet tails. Consistent with this, aggregation of an exclusively branched nucleation-promoting factor (the VCA domain of N-WASP), with density and turnover similar to those of N-WASP in Nck comets, did not reconstitute dynamic, elongated actin comets. Furthermore, enhancement of branched Arp2/3-mediated nucleation by N-WASP overexpression caused loss of the typical actin comet tail shape induced by Nck aggregation. Thus the ratio of linear to dendritic nucleation activity may serve to distinguish the properties of actin structures induced by various viral and bacterial pathogens. PMID:26609071

  20. The revised electromagnetic fields directive and worker exposure in environments with high magnetic flux densities.

    PubMed

    Stam, Rianne

    2014-06-01

    Some of the strongest electromagnetic fields (EMF) are found in the workplace. A European Directive sets limits to workers' exposure to EMF. This review summarizes its origin and contents and compares magnetic field exposure levels in high-risk workplaces with the limits set in the revised Directive. Pubmed, Scopus, grey literature databases, and websites of organizations involved in occupational exposure measurements were searched. The focus was on EMF with frequencies up to 10 MHz, which can cause stimulation of the nervous system. Selected studies had to provide individual maximum exposure levels at the workplace, either in terms of the external magnetic field strength or flux density or as induced electric field strength or current density. Indicative action levels and the corresponding exposure limit values for magnetic fields in the revised European Directive will be higher than those in the previous version. Nevertheless, magnetic flux densities in excess of the action levels for peripheral nerve stimulation are reported for workers involved in welding, induction heating, transcranial magnetic stimulation, and magnetic resonance imaging (MRI). The corresponding health effects exposure limit values for the electric fields in the worker's body can be exceeded for welding and MRI, but calculations for induction heating and transcranial magnetic stimulation are lacking. Since the revised European Directive conditionally exempts MRI-related activities from the exposure limits, measures to reduce exposure may be necessary for welding, induction heating, and transcranial nerve stimulation. Since such measures can be complicated, there is a clear need for exposure databases for different workplace scenarios with significant EMF exposure and guidance on good practices. PMID:24557933

  1. The Revised Electromagnetic Fields Directive and Worker Exposure in Environments With High Magnetic Flux Densities

    PubMed Central

    Stam, Rianne

    2014-01-01

    Some of the strongest electromagnetic fields (EMF) are found in the workplace. A European Directive sets limits to workers’ exposure to EMF. This review summarizes its origin and contents and compares magnetic field exposure levels in high-risk workplaces with the limits set in the revised Directive. Pubmed, Scopus, grey literature databases, and websites of organizations involved in occupational exposure measurements were searched. The focus was on EMF with frequencies up to 10 MHz, which can cause stimulation of the nervous system. Selected studies had to provide individual maximum exposure levels at the workplace, either in terms of the external magnetic field strength or flux density or as induced electric field strength or current density. Indicative action levels and the corresponding exposure limit values for magnetic fields in the revised European Directive will be higher than those in the previous version. Nevertheless, magnetic flux densities in excess of the action levels for peripheral nerve stimulation are reported for workers involved in welding, induction heating, transcranial magnetic stimulation, and magnetic resonance imaging (MRI). The corresponding health effects exposure limit values for the electric fields in the worker’s body can be exceeded for welding and MRI, but calculations for induction heating and transcranial magnetic stimulation are lacking. Since the revised European Directive conditionally exempts MRI-related activities from the exposure limits, measures to reduce exposure may be necessary for welding, induction heating, and transcranial nerve stimulation. Since such measures can be complicated, there is a clear need for exposure databases for different workplace scenarios with significant EMF exposure and guidance on good practices. PMID:24557933

  2. Pushing with actin: from cells to pathogens.

    PubMed

    Small, J Victor

    2015-02-01

    Actin polymerization is harnessed by cells to generate lamellipodia for movement and by a subclass of pathogens to facilitate invasion of their infected hosts. Using electron tomography (ET), we have shown that lamellipodia are formed via the generation of subsets of actin filaments joined by branch junctions. Image averaging produced a 2.9 nm resolution model of branch junctions in situ and revealed a close fit to the electron density map of the actin-related protein 2/3 (Arp2/3)-actin complex in vitro. Correlated live-cell imaging and ET was also used to determine how actin networks are created and remodelled during the initiation and inhibition of protrusion in lamellipodia. Listeria, Rickettsia and viruses, such as vaccinia virus and baculovirus, exploit the actin machinery of host cells to generate propulsive actin comet tails to disseminate their infection. By applying ET, we have shown that baculovirus generates at its rear a fishbone-like array of subsets of branched actin filaments, with an average of only four filaments engaged in pushing at any one time. In both of these studies, the application of ET of negatively stained cytoskeletons for higher filament resolution and cryo-ET for preserving overall 3D morphology was crucial for obtaining a complete structure-function analysis of actin-driven propulsion. PMID:25619250

  3. First-flight escape from spheres with R(-2) density distribution. [particle flux from comets, stars and unconfined plasmas

    NASA Technical Reports Server (NTRS)

    Huebner, W. F.; Keady, J. J.

    1984-01-01

    Energy-independent first-flight transport kernels are evaluated for a spherical region with an R(-2) density distribution. The uncollided angular-flux distribution is obtained and integrated for a source distribution that is proportional to the density to give the uncollided emitted particle flux and current density. These are useful for the calculation of mass, energy, and momentum carried away by fast particles born in the medium. The data are relevant to estimate escape from weakly bound atmospheres such as comet comae, dilute circumstellar envelopes, and some unconfined laboratory plasmas.

  4. Bacterial lipopolysaccharide induces actin reorganization, intercellular gap formation, and endothelial barrier dysfunction in pulmonary vascular endothelial cells: concurrent F-actin depolymerization and new actin synthesis.

    PubMed

    Goldblum, S E; Ding, X; Brann, T W; Campbell-Washington, J

    1993-10-01

    Bacterial lipopolysaccharide (LPS) influences pulmonary vascular endothelial barrier function in vitro. We studied whether LPS regulates endothelial barrier function through actin reorganization. Postconfluent bovine pulmonary artery endothelial cell monolayers were exposed to Escherichia coli 0111:B4 LPS 10 ng/ml or media for up to 6 h and evaluated for: 1) transendothelial 14C-albumin flux, 2) F-actin organization with fluorescence microscopy, 3) F-actin quantitation by spectrofluorometry, and 4) monomeric G-actin levels by the DNAse 1 inhibition assay. LPS induced increments in 14C-albumin flux (P < 0.001) and intercellular gap formation at > or = 2-6 h. During this same time period the endothelial F-actin pool was not significantly changed compared to simultaneous media controls. Mean (+/- SE) G-actin (micrograms/mg total protein) was significantly (P < 0.002) increased compared to simultaneous media controls at 2, 4, and 6 h but not at 0.5 or 1 h. Prior F-actin stabilization with phallicidin protected against the LPS-induced increments in G-actin (P = 0.040) as well as changes in barrier function (P < 0.0001). Prior protein synthesis inhibition unmasked an LPS-induced decrement in F-actin (P = 0.0044), blunted the G-actin increment (P = 0.010), and increased LPS-induced changes in endothelial barrier function (P < 0.0001). Therefore, LPS induces pulmonary vascular endothelial F-actin depolymerization, intercellular gap formation, and barrier dysfunction. Over the same time period, LPS increased total actin (P < 0.0001) and new actin synthesis (P = 0.0063) which may be a compensatory endothelial cell response to LPS-induced F-actin depolymerization. PMID:8408232

  5. An Approximate Analytic Expression for the Flux Density of Scintillation Light at the Photocathode

    SciTech Connect

    Braverman, Joshua B; Harrison, Mark J; Ziock, Klaus-Peter

    2012-01-01

    The flux density of light exiting scintillator crystals is an important factor affecting the performance of radiation detectors, and is of particular importance for position sensitive instruments. Recent work by T. Woldemichael developed an analytic expression for the shape of the light spot at the bottom of a single crystal [1]. However, the results are of limited utility because there is generally a light pipe and photomultiplier entrance window between the bottom of the crystal and the photocathode. In this study, we expand Woldemichael s theory to include materials each with different indices of refraction and compare the adjusted light spot shape theory to GEANT 4 simulations [2]. Additionally, light reflection losses from index of refraction changes were also taken into account. We found that the simulations closely agree with the adjusted theory.

  6. ATLBS EXTENDED SOURCE SAMPLE: THE EVOLUTION IN RADIO SOURCE MORPHOLOGY WITH FLUX DENSITY

    SciTech Connect

    Saripalli, L.; Subrahmanyan, R.; Thorat, K.; Ekers, R. D.; Hunstead, R. W.; Johnston, H. M.; Sadler, E. M.

    2012-04-01

    Based on the Australia Telescope Low Brightness Survey (ATLBS) we present a sample of extended radio sources and derive morphological properties of faint radio sources. One hundred nineteen radio galaxies form the ATLBS Extended Source Sample (ATLBS-ESS) consisting of all sources exceeding 30'' in extent and integrated flux densities exceeding 1 mJy. We give structural details along with information on galaxy identifications and source classifications. The ATLBS-ESS, unlike samples with higher flux-density limits, has almost equal fractions of FR-I and FR-II radio galaxies, with a large fraction of the FR-I population exhibiting 3C31-type structures. Significant asymmetry in lobe extents appears to be a common occurrence in the ATLBS-ESS FR-I sources compared with FR-II sources. We present a sample of 22 FR-Is at z > 0.5 with good structural information. The detection of several giant radio sources, with size exceeding 0.7 Mpc, at z > 1 suggests that giant radio sources are not less common at high redshifts. The ESS also includes a sample of 28 restarted radio galaxies. The relative abundance of dying and restarting sources is indicative of a model where radio sources undergo episodic activity in which an active phase is followed by a brief dying phase that terminates with restarting of the central activity; in any massive elliptical a few such activity cycles wherein adjacent events blend may constitute the lifetime of a radio source and such bursts of blended activity cycles may be repeated over the age of the host. The ATLBS-ESS includes a 2 Mpc giant radio galaxy with the lowest surface brightness lobes known to date.

  7. The effect of an on-orbit near encounter on the number flux density of micron sized particles

    NASA Technical Reports Server (NTRS)

    Maag, Carl R.; Tanner, William G.; Stevenson, Tim J.; Borg, Janet; Bibring, Jean-Pierre; Alexander, W. Merle; Maag, Andrew J.

    1993-01-01

    Many materials and techniques have been developed by the authors to sample the flux of particles in Low Earth Orbit (LEO), and through regular insitu sampling of the flux in LEO, the materials and techniques have produced data which complement the data now being amassed by the Long Duration Exposure Facility (LDEF) research activities. Recent flight experiments on STS-32, STS-44, STS-46, and STS-52 have been conducted to develop an understanding of the spatial density as a function of size (mass) for particle sizes 1x10(exp -6) cm and larger. In addition to the enumeration of particle impacts, it was also the intent of these experiments that hypervelocity particles be captured and returned intact. Measurements were performed post-flight to determine the flux density, diameters, and subsequent effects on various optical, thermal control, and structural materials. During the course of the STS-44 mission, the Space Shuttle corrected its altitude by 26 km to evade a spent upper stage. The results of this near encounter suggests that a cloud of micron sized particles exist in the vicinity of the object. Data also suggest that the flux density is nearly two (2) orders of magnitude higher than background flux. A comparison of the number flux density along with microphotographs of the captured particles will be presented for the referenced shuttle flights.

  8. A model of heat transfer in sapwood and implications for sap flux density measurements using thermal dissipation probes

    SciTech Connect

    Wullschleger, Stan D; Childs, Kenneth W; King, Anthony Wayne; Hanson, Paul J

    2011-01-01

    A variety of thermal approaches are used to estimate sap flux density in stems of woody plants. Models have proven valuable tools for interpreting the behavior of heat pulse, heat balance, and heat field deformation techniques, but have seldom been used to describe heat transfer dynamics for the heat dissipation method. Therefore, to better understand the behavior of heat dissipation probes, a model was developed that takes into account the thermal properties of wood, the physical dimensions and thermal characteristics of the probes, and the conductive and convective heat transfer that occurs due to water flow in the sapwood. Probes were simulated as aluminum tubes 20 mm in length and 2 mm in diameter, whereas sapwood, heartwood, and bark each had a density and water fraction that determined their thermal properties. Base simulations assumed a constant sap flux density with sapwood depth and no wounding or physical disruption of xylem beyond the 2 mm diameter hole drilled for probe installation. Simulations across a range of sap flux densities showed that the dimensionless quantity k defined as ( Tm T)/ T where Tm is the temperature differential ( T) between the heated and unheated probe under zero flow conditions was dependent on the thermal conductivity of the sapwood. The relationship between sap flux density and k was also sensitive to radial gradients in sap flux density and to xylem disruption near the probe. Monte Carlo analysis in which 1000 simulations were conducted while simultaneously varying thermal conductivity and wound diameter revealed that sap flux density and k showed considerable departure from the original calibration equation used with this technique. The departure was greatest for abrupt patterns of radial variation typical of ring-porous species. Depending on the specific combination of thermal conductivity and wound diameter, use of the original calibration equation resulted in an 81% under- to 48% over-estimation of sap flux density at

  9. Density of Gadolinium Nitrate Solutions for the High Flux Isotope Reactor

    SciTech Connect

    Taylor, Paul Allen; Lee, Denise L

    2009-05-01

    In late 1992, the High Flux Isotope Reactor (HFIR) was planning to switch the solution contained in the poison injection tank from cadmium nitrate to gadolinium nitrate. The poison injection system is an emergency system used to shut down the reactor by adding a neutron poison to the cooling water. This system must be able to supply a minimum of 69 pounds of gadolinium to the reactor coolant system in order to guarantee that the reactor would become subcritical. A graph of the density of gadolinium nitrate solutions over a concentration range of 5 to 30 wt% and a temperature range of 15 to 40{sup o}C was prepared. Routine density measurements of the solution in the poison injection tank are made by HFIR personnel, and an adaptation of the original graph is used to determine the gadolinium nitrate concentration. In late 2008, HFIR personnel decided that the heat tracing that was present on the piping for the poison injection system could be removed without any danger of freezing the solution; however, the gadolinium nitrate solution might get as cold as 5{sup o}C. This was outside the range of the current density-concentration correlation, so the range needed to be expanded. This report supplies a new density-concentration correlation that covers the extended temperature range. The correlation is given in new units, which greatly simplifies the calculation that is required to determine the pounds of gadolinium in the tank solution. The procedure for calculating the amount of gadolinium in the HFIR poison injection system is as follows: (1) Calculate the usable volume in the system; (2) Measure the density of the solution; (3) Calculate the gadolinium concentration using the following equation: Gd(lb/ft{sup 3}) = measured density (g/mL) x 34.681 - 34.785; (4) Calculate the amount of gadolinium in the system using the following equation: Amount of Gd(lb) = Gd concentration (lb/ft{sup 3}) x usable volume (ft{sup 3}). The equation in step 3 is exact for a temperature of

  10. An investigation into the torque density capabilities of flux-focusing magnetic gearboxes

    NASA Astrophysics Data System (ADS)

    Uppalapati, Krishna Kiran

    Wind and many rotary based ocean energy conversion devices rely on a mechanical gearbox to increase their speed so as to match the requirements of the electromagnetic generator. However, mechanical gearboxes have a number of disadvantages such as the need for gear lubrication, no overload protection and the creation of acoustic noise. Frequently direct-drive generators are employed to overcome these issues, wherein the gearbox is removed and the shaft of the turbine is directly connected to the synchronous generator, either with an electrically excited or permanent magnet rotor. If the input speed to the generator is very low the torque must be very high in order to generate the necessary power. However, as the electrical loading of a synchronous generator is thermally limited, the size of the generator will become excessively large at high power levels. An alternative to these technologies is to consider replacing the mechanical gearbox with a magnetic gear. A magnetic gear can create speed change without any physical contact. It has inherent overload protection, and its non-contact operation offers the potential for high reliability. Despite significant progress, existing magnetic gear designs do not achieve torque densities that are competitive with mechanical gearboxes. This research has focused on designing a coaxial magnetic gear that can operate at a volumetric torque density that is comparable to a mechanical gearbox. A flux-focusing rotor topology also called spoke-type rotor magnet arrangement was adopted to improve the air-gap magnetic flux density which in turn improves the torque transferred between the rotors. Finite element analysis was utilized to conduct a parameter sweep analysis of the different geometric parameters of the magnetic gear. A sub-scale magnetic gear with a diameter of 110 mm and a scaled-up magnetic gear with a diameter of 228 mm was designed, constructed and experimentally evaluated. The torque and torque density of sub

  11. Dynamics of photosynthetic photon flux density (PPFD) and estimates in coastal northern California

    NASA Astrophysics Data System (ADS)

    Ge, Shaokui; Smith, Richard G.; Jacovides, Constantinos P.; Kramer, Marc G.; Carruthers, Raymond I.

    2011-08-01

    Plants require solar radiation for photosynthesis and their growth is directly related to the amount received, assuming that other environmental parameters are not limiting. Therefore, precise estimation of photosynthetically active radiation (PAR) is necessary to enhance overall accuracies of plant growth models. This study aimed to explore the PAR radiant flux in the San Francisco Bay Area of northern California. During the growing season (March through August) for 2 years 2007-2008, the on-site magnitudes of photosynthetic photon flux densities (PPFD) were investigated and then processed at both the hourly and daily time scales. Combined with global solar radiation ( R S) and simulated extraterrestrial solar radiation, five PAR-related values were developed, i.e., flux density-based PAR (PPFD), energy-based PAR (PARE), from-flux-to-energy conversion efficiency (fFEC), and the fraction of PAR energy in the global solar radiation (fE), and a new developed indicator—lost PARE percentages (LPR)—when solar radiation penetrates from the extraterrestrial system to the ground. These PAR-related values indicated significant diurnal variation, high values occurring at midday, with the low values occurring in the morning and afternoon hours. During the entire experimental season, the overall mean hourly value of fFEC was found to be 2.17 μmol J-1, while the respective fE value was 0.49. The monthly averages of hourly fFEC and fE at the solar noon time ranged from 2.15 in March to 2.39 μmol J-1 in August and from 0.47 in March to 0.52 in July, respectively. However, the monthly average daily values were relatively constant, and they exhibited a weak seasonal variation, ranging from 2.02 mol MJ-1 and 0.45 (March) to 2.19 mol MJ-1 and 0.48 (June). The mean daily values of fFEC and fE at the solar noon were 2.16 mol MJ-1 and 0.47 across the entire growing season, respectively. Both PPFD and the ever first reported LPR showed strong diurnal patterns. However, they had

  12. Low Temperature Nitriding of 304 Austenitic Stainless Steel Using RF-ICP Method: the Role of Ion Beam Flux Density

    NASA Astrophysics Data System (ADS)

    Wang, Qing; Ba, Dechun; Ming, Yue; Xu, Lin; Guo, Deyu

    2014-10-01

    The significant role of ion beam flux during nitriding 304 austenitic stainless steel has been investigated by using a radio frequency inductively-coupled plasma reactor into which a sample with negative bias voltage was inserted. A milliammeter is used to detect the current of ions which collide with the sample and optical emission spectroscopy is used to discern the reactive species included in the nitrogen plasma. The nitriding efficiency is indicated by X-ray diffraction and the microhardness test. The reported data reveal that the ion beam flux density as well as the deposition pressure, bias voltage and time can strongly affect the nitriding of stainless steel via the expanded multiphase microstructure inside the nitrided layer. The increase in the density of ion flux results in an ascent in the intensity of the expanded peak and a simultaneous decline in the intensity of the γ austenite peak. The evolution trend of ion beam flux density is described as a function of the operating pressure and the bias voltage. The maximum ion flux density has been achieved at 10 Pa pressure and -500 V bias voltage. A reasonable nitriding region has been, consequently, suggested after comparing this work with previously reported results.

  13. VizieR Online Data Catalog: PKS 0405-385 flux-density monitoring (Kedziora-Chudczer+, 2006)

    NASA Astrophysics Data System (ADS)

    Kedziora-Chudczer, L.

    2007-01-01

    The monitoring of PKS 0405-385 commenced in 1993 November, as part of the ATCA IDV Survey (Kedziora-Chudczer et al., 2001, Cat. ) with the Australia Telescope Compact Array.1 The source showed rapid variability in total flux density at 8.6, 4.8 and 2.4GHz during these observations. The time-scale of variability, defined throughout this paper as HWHP of auto-correlation function of flux density, was faster than 2h (the frequency of sampling used in the survey). (2 data files).

  14. Analysis of the relationship between photosynthetic photon flux density and natural Taxus baccata seedlings occurrence

    NASA Astrophysics Data System (ADS)

    Iszkuło, Grzegorz; Boratyński, Adam

    2006-01-01

    The aim of the present work was to analyse the relationship of seedlings and saplings of Taxus baccata to the photosynthetic photon flux density (PPFD) reaching the forest floor under natural conditions. Two permanent plots, subdivided into 1 × 1 m square plots, were established in a naturally regenerating population of T. baccata formed during last decades in the Kórnik Arboretum, Poland. All seedlings in every 1 × 1 m plots were counted. Relative PPFD was measured for every plot at the canopy height of the yew seedlings. The dependence of seedling density upon PPFD was examined. We found, that the frequency of the smallest seedlings (to 6.0 cm tall) was highest in the most shaded plots and decreased in plots with increasing PPFD. Thus, the youngest yew seedlings can germinate and grow in very shady conditions. However, the older seedlings (6.1-25.0 and 25.1-100.0 classes) were observed most frequently in 2-7% PPFD. The small numbers of older, taller seedlings in deep shade likely indicate a higher mortality rate of seedlings less than 6 cm in height without promotion to the next height class. Probably the low value of PPFD under the canopy of the stand significantly reduces the competition of other plants with the youngest yew seedlings. At higher light levels they may not be able to compete with more light-demanding plants, such as herbs and seedlings of broad-leaved trees. The seedlings of the second (6.1-25.0 cm) and third (25.1-100.0 cm) height classes were observed most frequently in the plots with 2-7% PPFD ( Fig. 1b and c).

  15. Observations of Poynting fluxes, ion temperatures and neutral densities during the March 2015 magnetic storm

    NASA Astrophysics Data System (ADS)

    Huang, Y.; Su, Y. J.; Huang, C. Y.; Hairston, M. R.; Sutton, E. K.

    2015-12-01

    We will present various observations regarding the geomagnetic energy input and the response of Ionosphere-Thermosphere (IT) system during the March 17, 2015 storm, the largest one in solar cycle 24. The Poynting fluxes measured by Defense Meteorological Satellite Program (DMSP) satellites (F16, F17 and F18) show significant enhancements in the auroral oval and at high latitudes poleward of the auroral oval. Moreover, the ion temperatures observed by DMSP satellites (F16, F17 and F19) at magnetic latitudes greater than 80° are higher than those in the auroral oval, and the their averaged increases are 316K in the northern hemisphere and 248 K in the southern hemisphere, respectively. In addition, the neutral density residuals measured by the Gravity Recovery and Climate Experiment (GRACE) satellite indicate the largest values at the highest orbital latitudes. The wave-like perturbations originating at high latitudes move equatorward with decreasing amplitudes along GRACE orbits, implying a source region for Traveling Atmospheric Disturbances (TADs) at polar latitudes.

  16. Industrialization of nanocrystalline Fe-Si-B-P-Cu alloys for high magnetic flux density cores

    NASA Astrophysics Data System (ADS)

    Takenaka, Kana; Setyawan, Albertus D.; Sharma, Parmanand; Nishiyama, Nobuyuki; Makino, Akihiro

    2016-03-01

    Nanocrystalline Fe-Si-B-P-Cu alloys exhibit high saturation magnetic flux density (Bs) and extremely low magnetic core loss (W), simultaneously. Low amorphous-forming ability of these alloys hinders their application potential in power transformers and motors. Here we report a solution to this problem. Minor addition of C is found to be effective in increasing the amorphous-forming ability of Fe-Si-B-P-Cu alloys. It allows fabrication of 120 mm wide ribbons (which was limited to less than 40 mm) without noticeable degradation in magnetic properties. The nanocrystalline (Fe85.7Si0.5B9.5P3.5Cu0.8)99C1 ribbons exhibit low coercivity (Hc)~4.5 A/m, high Bs~1.83 T and low W~0.27 W/kg (@ 1.5 T and 50 Hz). Success in fabrication of long (60-100 m) and wide (~120 mm) ribbons, which are made up of low cost elements is promising for mass production of energy efficient high power transformers and motors

  17. Excessive magnetic field flux density distribution from overhead isolated powerline conductors due to neutral line current.

    PubMed

    Netzer, Moshe

    2013-06-01

    Overhead isolated powerline conductors (hereinafter: "OIPLC") are the most compact form for distributing low voltage currents. From the known physics of magnetic field emission from 3-phase power lines, it is expected that excellent symmetry of the 120° shifted phase currents and where compact configuration of the 3-phase+neutral line exist, the phase current vectorial summation of the magnetic field flux density (MFFD) is expected to be extremely low. However, despite this estimation, an unexpectedly very high MFFD was found in at least three towns in Israel. This paper explains the reasons leading to high MFFD emissions from compact OIPLC and the proper technique to fix it. Analysis and measurement results had led to the failure hypothsis of neutral line poor connection design and poor grounding design of the HV-LV utility transformers. The paper elaborates on the low MFFD exposure level setup by the Israeli Environmental Protection Office which adopted a rather conservative precaution principal exposure level (2 mG averaged over 24 h). PMID:23675630

  18. Absolute flux density calibrations of radio sources: 2.3 GHz

    NASA Technical Reports Server (NTRS)

    Freiley, A. J.; Batelaan, P. D.; Bathker, D. A.

    1977-01-01

    A detailed description of a NASA/JPL Deep Space Network program to improve S-band gain calibrations of large aperture antennas is reported. The program is considered unique in at least three ways; first, absolute gain calibrations of high quality suppressed-sidelobe dual mode horns first provide a high accuracy foundation to the foundation to the program. Second, a very careful transfer calibration technique using an artificial far-field coherent-wave source was used to accurately obtain the gain of one large (26 m) aperture. Third, using the calibrated large aperture directly, the absolute flux density of five selected galactic and extragalactic natural radio sources was determined with an absolute accuracy better than 2 percent, now quoted at the familiar 1 sigma confidence level. The follow-on considerations to apply these results to an operational network of ground antennas are discussed. It is concluded that absolute gain accuracies within + or - 0.30 to 0.40 db are possible, depending primarily on the repeatability (scatter) in the field data from Deep Space Network user stations.

  19. Influence of stem temperature changes on heat pulse sap flux density measurements.

    PubMed

    Vandegehuchte, Maurits W; Burgess, Stephen S O; Downey, Alec; Steppe, Kathy

    2015-04-01

    While natural spatial temperature gradients between measurement needles have been thoroughly investigated for continuous heat-based sap flow methods, little attention has been given to how natural changes in stem temperature impact heat pulse-based methods through temporal rather than spatial effects. By modelling the theoretical equation for both an ideal instantaneous pulse and a step pulse and applying a finite element model which included actual needle dimensions and wound effects, the influence of a varying stem temperature on heat pulse-based methods was investigated. It was shown that the heat ratio (HR) method was influenced, while for the compensation heat pulse and Tmax methods changes in stem temperatures of up to 0.002 °C s(-1) did not lead to significantly different results. For the HR method, rising stem temperatures during measurements led to lower heat pulse velocity values, while decreasing stem temperatures led to both higher and lower heat pulse velocities, and to imaginary results for high flows. These errors of up to 40% can easily be prevented by including a temperature correction in the data analysis procedure, calculating the slope of the natural temperature change based on the measured temperatures before application of the heat pulse. Results of a greenhouse and outdoor experiment on Pinus pinea L. show the influence of this correction on low and average sap flux densities. PMID:25145698

  20. Leaf photosynthetic and solar-tracking responses of mallow, Malva parviflora, to photon flux density.

    PubMed

    Greer, Dennis H; Thorpe, Michael R

    2009-10-01

    Malva parviflora L. (mallow) is a species that occupies high-light habitats as a weedy invader in orchards and vineyards. Species of the Malvaceae are known to solar track and anecdotal evidence suggests this species may also. How M. parviflora responds physiologically to light in comparison with other species within the Malvaceae remains unknown. Tracking and photosynthetic responses to photon flux density (PFD) were evaluated on plants grown in greenhouse conditions. Tracking ability was assessed in the growth conditions and by exposing leaves to specific light intensities and measuring changes in the angle of the leaf plane. Light responses were also determined by photosynthesis and chlorophyll fluorescence. Leaves followed a heliotropic response which was highly PFD-dependent, with tracking rates increasing in a curvilinear pattern. Maximum tracking rates were up to 20 degrees h(-1) and saturated for light above 1,300 micromol (photons) m(-2) s(-1). This high-light saturation, both for tracking (much higher than the other species), and for photosynthesis, confirmed mallow as a high-light demanding species. Further, because there was no photoinhibition, the leaves could capture the potential of an increased carbon gain in higher irradiance by resorting to solar tracking. Modelling suggested the tracking response could increase the annual carbon gain by as much as 25% compared with leaves that do not track the sun. The various leaf attributes associated with solar tracking, therefore, help to account for the success of this species as a weed in many locations worldwide. PMID:19576789

  1. Photosynthetic photon flux density, carbon dioxide concentration, and vapor pressure deficit effects on photosynthesis in cacao seedlings

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cacao (Theobroma cacao) is a shade plant, native to the under-story of the evergreen rain forest of the Amazon basin and adapted to low levels of photosynthetic photon flux density (PPFD). The influence of PPFD, leaf to air water vapor pressure deficit (VPD) and external carbon dioxide concentration...

  2. Actin in Herpesvirus Infection

    PubMed Central

    Roberts, Kari L.; Baines, Joel D.

    2011-01-01

    Actin is important for a variety of cellular processes, including uptake of extracellular material and intracellular transport. Several emerging lines of evidence indicate that herpesviruses exploit actin and actin-associated myosin motors for viral entry, intranuclear transport of capsids, and virion egress. The goal of this review is to explore these processes and to highlight potential future directions for this area of research. PMID:21994736

  3. Evidence of Short Timescale Flux Density Variations of UC HII Regions in Sgr B2 Main and North

    NASA Astrophysics Data System (ADS)

    De Pree, C. G.; Peters, T.; Mac Low, M. M.; Wilner, D. J.; Goss, W. M.; Galván-Madrid, R.; Keto, E. R.; Klessen, R. S.; Monsrud, A.

    2015-12-01

    We have recently published observations of significant flux density variations at 1.3 cm in H ii regions in the star-forming regions Sgr B2 Main and North. To further study these variations, we have made new 7 mm continuum and recombination line observations of Sgr B2 at the highest possible angular resolution of the Karl G. Jansky Very Large Array (VLA). We have observed Sgr B2 Main and North at 42.9 GHz and at 45.4 GHz in the BnA configuration (Main) and the A configuration (North). We compare these new data to archival VLA 7 mm continuum data of Sgr B2 Main observed in 2003 and Sgr B2 North observed in 2001. We find that 1 of the 41 known ultracompact and hypercompact H ii regions in Sgr B2 (K2-North) has decreased ∼27% in flux density from 142 ± 14 to 103 ± 10 mJy (2.3σ) between 2001 and 2012. A second source, F3c-Main, has increased ∼30% in flux density from 82 ± 8 to 107 ± 11 mJy (1.8σ) between 2003 and 2012. F3c-Main was previously observed to increase in flux density at 1.3 cm over a longer time period between 1989 and 2012. An observation of decreasing flux density, such as that observed in K2-North, is particularly significant since such a change is not predicted by the classical hypothesis of steady expansion of H ii regions during massive star accretion. Our new observations at 7 mm, along with others in the literature, suggest that the formation of massive stars occurs through time-variable and violent accretion.

  4. The turbulent heat flux in low Mach number flows with large density variations

    NASA Technical Reports Server (NTRS)

    Orourke, Peter J.; Collins, Lance R.

    1988-01-01

    A transport equation has been derived which is the difference between the volume- and mass-averaged velocities and is simply related to the turbulent heat flux phi sup h. Using this equation and an assumption analogous to the drift flux approximation of two-phase flow modeling, an algebraic closure relation for phi sup h that exibits fluxes due to directed transport proportional to -del anti p and due to gradient transport proportional to -del tau has been obtained.

  5. Actin from Saccharomyces cerevisiae.

    PubMed Central

    Greer, C; Schekman, R

    1982-01-01

    Inhibition of DNase I activity has been used as an assay to purify actin from Saccharomyces cerevisiae (yeast actin). The final fraction, obtained after a 300-fold purification, is approximately 97% pure as judged by sodium dodecyl sulfate-gel electrophoresis. Like rabbit skeletal muscle actin, yeast actin has a molecular weight of about 43,000, forms 7-nm-diameter filaments when polymerization is induced by KCl or Mg2+, and can be decorated with a proteolytic fragment of muscle myosin (heavy meromyosin). Although heavy meromyosin ATPase activity is stimulated by rabbit muscle and yeast actins to approximately the same Vmax (2 mmol of Pi per min per mumol of heavy meromyosin), half-maximal activation (Kapp) is obtained with 14 micro M muscle actin, but requires approximately 135 micro M yeast actin. This difference suggests a low affinity of yeast actin for muscle myosin. Yeast and muscle filamentous actin respond similarly to cytochalasin and phalloidin, although the drugs have no effect on S. cerevisiae cell growth. Images PMID:6217414

  6. Actin Rings of Power.

    PubMed

    Schwayer, Cornelia; Sikora, Mateusz; Slováková, Jana; Kardos, Roland; Heisenberg, Carl-Philipp

    2016-06-20

    Circular or ring-like actin structures play important roles in various developmental and physiological processes. Commonly, these rings are composed of actin filaments and myosin motors (actomyosin) that, upon activation, trigger ring constriction. Actomyosin ring constriction, in turn, has been implicated in key cellular processes ranging from cytokinesis to wound closure. Non-constricting actin ring-like structures also form at cell-cell contacts, where they exert a stabilizing function. Here, we review recent studies on the formation and function of actin ring-like structures in various morphogenetic processes, shedding light on how those different rings have been adapted to fulfill their specific roles. PMID:27326928

  7. Data-Model Comparison Investigations of Thermospheric Density and Composition Influences on High-Altitude Photoelectron Fluxes at Mars

    NASA Astrophysics Data System (ADS)

    Xu, S.; Liemohn, M. W.; Bougher, S. W.; Mitchell, D. L.

    2014-12-01

    We present results from a superthermal electron transport code adapted for the Mars environment to study the controlling factors of high-altitude/escaped photoelectron fluxes at this planet. In addition to numerical checks of the code, we investigate the influences of the following effects: magnetic field configuration, solar EUV flux input, and atmospheric density/temperature profiles. In particular, we explore the causes of the extremely high photoelectron fluxes, resulting into two linear dependent trends on solar EUV proxy, measured by the Mars Global Surveyor MAGnetometer/Electron Reflector (MGS MAG/ER) in late 2001-early 2002 (Mars year 25). Studies have shown the relation between these high fluxes and the global dust storm that occurred in the same time period. This modeling work further explores the physical explanations of this relation. Our preliminary results suggest that an increase in CO2 density in the upper thermosphere (150-300 km altitude) is necessary to match the observed changes in photoelectron flux at the MGS altitude of ~400 km.

  8. A dynamo theory prediction for solar cycle 22: Sunspot number, radio flux, exospheric temperature, and total density at 400 km

    NASA Technical Reports Server (NTRS)

    Schatten, K. H.; Hedin, A. E.

    1986-01-01

    Using the dynamo theory method to predict solar activity, a value for the smoothed sunspot number of 109 + or - 20 is obtained for solar cycle 22. The predicted cycle is expected to peak near December, 1990 + or - 1 year. Concommitantly, F(10.7) radio flux is expected to reach a smoothed value of 158 + or - 18 flux units. Global mean exospheric temperature is expected to reach 1060 + or - 50 K and global total average total thermospheric density at 400 km is expected to reach 4.3 x 10 to the -15th gm/cu cm + or - 25 percent.

  9. A dynamo theory prediction for solar cycle 22 - Sunspot number, radio flux, exospheric temperature, and total density at 400 km

    NASA Technical Reports Server (NTRS)

    Schatten, K. H.; Hedin, A. E.

    1984-01-01

    Using the 'dynamo theory' method to predict solar activity, a value for the smoothed sunspot number of 109 + or - 20 is obtained for solar cycle 22. The predicted cycle is expected to peak near December, 1990 + or - 1 year. Concommitantly, F(10.7) radio flux is expected to reach a smoothed value of 158 + or - 18 flux units. Global mean exospheric temperature is expected to reach 1060 + or - 50 K and global total average total thermospheric density at 400 km is expected to reach 4.3 x 10 to the -15th gm/cu cm + or - 25 percent.

  10. Effect of a magnetic flux line on the quantum beats in the Henon-Heiles level density.

    PubMed

    Brack, M.; Bhaduri, R. K.; Law, J.; Maier, Ch.; Murthy, M. V. N.

    1995-03-01

    The quantum density of states of the Henon-Heiles potential displays a pronounced beating pattern. This has been explained by the interference of three isolated classical periodic orbits with nearby actions and periods. A singular magnetic flux line, passing through the origin, drastically alters the beats even though the classical Lagrangian equations of motion remain unchanged. Some of the changes can be easily understood in terms of the Aharonov-Bohm effect. However, we find that the standard periodic orbit theory does not reproduce the diffraction-like quantum effects on those classical orbits which intersect the singular flux line, and argue that corrections of relative order variant Planck's over 2pi are necessary to describe these effects. We also discuss the changes in the distribution of nearest-neighbor spacings in the eigenvalue spectrum, brought about by the flux line. (c) 1995 American Institute of Physics. PMID:12780185

  11. Direct dynamin–actin interactions regulate the actin cytoskeleton

    PubMed Central

    Gu, Changkyu; Yaddanapudi, Suma; Weins, Astrid; Osborn, Teresia; Reiser, Jochen; Pollak, Martin; Hartwig, John; Sever, Sanja

    2010-01-01

    The large GTPase dynamin assembles into higher order structures that are thought to promote endocytosis. Dynamin also regulates the actin cytoskeleton through an unknown, GTPase-dependent mechanism. Here, we identify a highly conserved site in dynamin that binds directly to actin filaments and aligns them into bundles. Point mutations in the actin-binding domain cause aberrant membrane ruffling and defective actin stress fibre formation in cells. Short actin filaments promote dynamin assembly into higher order structures, which in turn efficiently release the actin-capping protein (CP) gelsolin from barbed actin ends in vitro, allowing for elongation of actin filaments. Together, our results support a model in which assembled dynamin, generated through interactions with short actin filaments, promotes actin polymerization via displacement of actin-CPs. PMID:20935625

  12. Effects of water salinity on the correlation scale of Root density and Evapotranspiration fluxes

    NASA Astrophysics Data System (ADS)

    Ajeel, Ali; Saeed, Ali; Dragonetti, Giovanna; Comegna, Alessandro; Lamaddalena, Nicola; Coppola, Antonio

    2015-04-01

    Spatial pattern and the correlation of different soil and plant parameters were examined in a green bean field experiment carried out at the Mediterranean Agronomic Institute of Bari, Italy. The experiment aimed to evaluate the role of local processes of salt accumulation and transport which mainly influences the evapotranspiration (and thus the root uptake) processes under different water salinity levels. The experiment consisted of three transects of 30m length and 4.2 m width, irrigated with three different salinity levels (1dSm-1, 3dSm-1, 6dSm-1). Soil measurements (electrical conductivity and soil water content) were monitored along transects in 24 sites, 1 m apart by using TDR probes and Diviner 2000. Water storage measured by TDR and Diviner sensor were coupled for calculating directly the evapotranspiration fluxes along the whole soil profile under the different salinity levels imposed during the experiment. In the same sites, crop monitoring involved measurements of Leaf Area Index (LAI), Osmotic Potential (OP), Leaf Water Potential (LWP), and Root length Density (RlD). Soil and plant properties were analyzed by classical statistics, geostatistics methods and spectral analysis. Results indicated moderate to large spatial variability across the field for soil and plant parameters under all salinity treatments. Furthermore, cross-semivariograms exhibited a strong positive spatial interdependence between electrical conductivity of soil solution ECw with ET and RlD in transect treated with 3dSm-1 as well as with LAI in transect treated with 6dSm-1 at all 24 monitoring sites. Spectral analysis enabled to identify the observation window to sample the soil salinity information responsible for a given plant response (ET, OP, RlD). It is also allowed a clear identification of the spatial scale at which the soil water salinity level and distribution and the crop response in terms of actual evapotranspiration ET, RlD and OP, are actually correlated. Additionally

  13. Carbon Gain and Photosynthetic Response of Chrysanthemum to Photosynthetic Photon Flux Density Cycles 1

    PubMed Central

    Stoop, Johan M. H.; Willits, Dan H.; Peet, Mary M.; Nelson, Paul V.

    1991-01-01

    Most models of carbon gain as a function of photosynthetic irradiance assume an instantaneous response to increases and decreases in irradiance. High- and low-light-grown plants differ, however, in the time required to adjust to increases and decreases in irradiance. In this study the response to a series of increases and decreases in irradiance was observed in Chrysanthemum × morifolium Ramat. “Fiesta” and compared with calculated values assuming an instantaneous response. There were significant differences between high- and low-light-grown plants in their photosynthetic response to four sequential photosynthetic photon flux density (PPFD) cycles consisting of 5-minute exposures to 200 and 400 micromoles per square meter per second (μmol m−2s−1). The CO2 assimilation rate of high-light-grown plants at the cycle peak increased throughout the PPFD sequence, but the rate of increase was similar to the increase in CO2 assimilation rate observed under continuous high-light conditions. Low-light leaves showed more variability in their response to light cycles with no significant increase in CO2 assimilation rate at the cycle peak during sequential cycles. Carbon gain and deviations from actual values (percentage carbon gain over- or underestimation) based on assumptions of instantaneous response were compared under continuous and cyclic light conditions. The percentage carbon gain overestimation depended on the PPFD step size and growth light level of the leaf. When leaves were exposed to a large PPFD increase, the carbon gain was overestimated by 16 to 26%. The photosynthetic response to 100 μmol m−2 s−1 PPFD increases and decreases was rapid, and the small overestimation of the predicted carbon gain, observed during photosynthetic induction, was almost entirely negated by the carbon gain underestimation observed after a decrease. If the PPFD cycle was 200 or 400 μmol m−2 s−1, high- and low-light leaves showed a carbon gain overestimation of 25

  14. Reverse propagation and negative angular momentum density flux of an optical nondiffracting nonparaxial fractional Bessel vortex beam of progressive waves.

    PubMed

    Mitri, F G

    2016-09-01

    Energy and angular momentum flux density characteristics of an optical nondiffracting nonparaxial vector Bessel vortex beam of fractional order are examined based on the dual-field method for the generation of symmetric electric and magnetic fields. Should some conditions determined by the polarization state, the half-cone angle as well as the beam-order (or topological charge) be met, the axial energy and angular momentum flux densities vanish (representing Poynting singularities), before they become negative. These negative counterintuitive properties suggest retrograde (negative) propagation as well as a rotation reversal of the angular momentum with respect to the beam handedness. These characteristics of nondiffracting nonparaxial Bessel fractional vortex beams of progressive waves open new capabilities in optical tractor beam tweezers, optical spanners, invisibility cloaks, optically engineered metamaterials, and other applications. PMID:27607486

  15. Milliarcsecond Change of IM Pegasi Radio Position in 1 Hour Coincident with Sharp Rise in Flux Density

    NASA Astrophysics Data System (ADS)

    Lebach, D. E.; Ratner, M. I.; Shapiro, I. I.; Ransom, R. R.; Bietenholz, M. F.; Bartel, N.; Lestrade, J.-F.

    1999-05-01

    Continuum VLBI observations at 3.6 cm of the RS CVn binary star IM Pegasi (HR 8703) for ~16 hr beginning on 1997 January 16 revealed an apparent motion of the star's radio position that coincided temporally with a large relative change in its flux density. Specifically, a rise in flux density from 18 to 46 mJy in 1.4 hr coincided with a detected position change over that interval of (Δα, Δδ)=(-0.68+/-0.15, 0.55+/-0.20) mas. The magnitude of this position change is much larger than can be explained by parallax, proper motion, and orbital motion and is about two-thirds the estimated angular diameter of the primary component of the binary.

  16. Spectrum and density of neutron flux in the irradiation beam line no. 3 of the IBR-2 reactor

    NASA Astrophysics Data System (ADS)

    Shabalin, E. P.; Verkhoglyadov, A. E.; Bulavin, M. V.; Rogov, A. D.; Kulagin, E. N.; Kulikov, S. A.

    2015-03-01

    Methodology and results of measuring the differential density of the neutron flux in irradiation beam line no. 3 of the IBR-2 reactor using neutron activation analysis (NAA) are presented in the paper. The results are compared to the calculation performed on the basis of the 3D MCNP model. The data that are obtained are required to determine the integrated radiation dose of the studied samples at various distances from the reactor.

  17. Measurement of the light flux density patterns from luminaires proposed as photon sources for photosynthesis during space travel

    NASA Technical Reports Server (NTRS)

    Walker, Paul N.

    1989-01-01

    Two luminaires were evaluated to determine the light flux density pattern on a horizontal plane surface. NASA supplied both luminaires; one was made by NASA and the other is commercially available. Tests were made for three combinations of luminaire height and luminaire lens material using the NASA luminaire; only one configuration of the commercial luminaire was tested. Measurements were made using four sensors with different wavelength range capabilities. The data are presented in graphical and tabular formats.

  18. A new measurement method of magnetic flux density using magnetorheological fluid characteristics and a variable resistor circuit

    NASA Astrophysics Data System (ADS)

    Kim, Hwan-Choong; Han, Chulhee; Kim, Pyunghwa; Choi, Seung-Bok

    2015-08-01

    This work proposes a new approach with which to measure the magnetic flux density using the characteristics of magnetorheological fluid (MRF) that is integrated with a variable resistor. For convenience, it is called a magnetorheological fluid variable resistor (MRF-VR) system in this study. The mechanism of the MRF-VR is based on the interaction between ferromagnetic iron particles of the MRF due to an external magnetic field, which causes its electrical resistance to be field dependent. Using this salient principle, the proposed MRF-VR system is constructed with electrodes and MRF, and its performance is demonstrated by evaluating its electrical resistive characteristics such as dimensional influence, response time, hysteresis and frequency response. After evaluating the performance characteristics, a feedback control system with a proportional-integral-derivative (PID) controller is established, and resistance-trajectory control experiments are carried out. Based on this MRF-VR system, a magnetic field-sensing system is constructed using a Wheatstone bridge circuit, and a polynomial model for calculating the magnetic flux density is formulated from the measured voltage. Finally, the accuracy and effectiveness of the proposed sensing system associated with the empirical polynomial model is successfully verified by comparing the calculated values of magnetic flux density with those measured by a commercial tesla meter.

  19. Actin Mechanics and Fragmentation*

    PubMed Central

    De La Cruz, Enrique M.; Gardel, Margaret L.

    2015-01-01

    Cell physiological processes require the regulation and coordination of both mechanical and dynamical properties of the actin cytoskeleton. Here we review recent advances in understanding the mechanical properties and stability of actin filaments and how these properties are manifested at larger (network) length scales. We discuss how forces can influence local biochemical interactions, resulting in the formation of mechanically sensitive dynamic steady states. Understanding the regulation of such force-activated chemistries and dynamic steady states reflects an important challenge for future work that will provide valuable insights as to how the actin cytoskeleton engenders mechanoresponsiveness of living cells. PMID:25957404

  20. Solar Modulation of Inner Trapped Belt Radiation Flux as a Function of Atmospheric Density

    NASA Technical Reports Server (NTRS)

    Lodhi, M. A. K.

    2005-01-01

    No simple algorithm seems to exist for calculating proton fluxes and lifetimes in the Earth's inner, trapped radiation belt throughout the solar cycle. Most models of the inner trapped belt in use depend upon AP8 which only describes the radiation environment at solar maximum and solar minimum in Cycle 20. One exception is NOAAPRO which incorporates flight data from the TIROS/NOAA polar orbiting spacecraft. The present study discloses yet another, simple formulation for approximating proton fluxes at any time in a given solar cycle, in particular between solar maximum and solar minimum. It is derived from AP8 using a regression algorithm technique from nuclear physics. From flux and its time integral fluence, one can then approximate dose rate and its time integral dose.

  1. A direct measurement of the energy flux density in plasma surface interaction

    NASA Astrophysics Data System (ADS)

    Dussart, Remi; Thomann, Anne-Lise; Semmar, Nadjib; Pichon, Laurianne; Bedra, Larbi; Mathias, Jacky; Tessier, Yves; Lefaucheux, Philippe

    2008-10-01

    The energy flux transferred from a plasma to a surface is a key issue for materials processing (sputtering, etching). We present direct measurements made with a Heat Flux Microsensor (HFM) in an Ar plasma interacting with the surface of the sensor. The HFM is a thermopile of about one thousand metal couples mounted in parallel. An Inductively Coupled Plasma in Argon was used to make the experiments. Langmuir probe and tuneable laser diode absorption measurements were carried out to estimate the contribution of ions, neutrals (conduction) and metastables. In order to evaluate the ability of the HFM to measure the part due to chemical reactions, a Si surface in contact with the HFM was submitted to an SF6 plasma. The direct measurements are in good agreement with the estimation we made knowing the etch rate and the enthalpy of the reaction. Finally, tests were performed on a sputtering reactor. Additional energy flux provided by condensing atoms (Pt) was also measured.

  2. Actin Polymerization is Stimulated by Actin Crosslinking Protein Palladin

    PubMed Central

    Gurung, Ritu; Yadav, Rahul; Brungardt, Joseph G.; Orlova, Albina; Egelman, Edward H.; Beck, Moriah R.

    2016-01-01

    The actin scaffold protein palladin regulates both normal cell migration and invasive cell motility, processes that require the coordinated regulation of actin dynamics. However, the potential effect of palladin on actin dynamics has remained elusive. Here we show that the actin binding immunoglobulin-like domain of palladin, which is directly responsible for both actin binding and bundling, also stimulates actin polymerization in vitro. Palladin eliminated the lag phase that is characteristic of the slow nucleation step of actin polymerization. Furthermore, palladin dramatically reduced depolymerization, slightly enhanced the elongation rate, and did not alter the critical concentration. Microscopy and in vitro crosslinking assays reveal differences in actin bundle architecture when palladin is incubated with actin before or after polymerization. These results suggest a model whereby palladin stimulates a polymerization-competent form of G-actin, akin to metal ions, either through charge neutralization or conformational changes. PMID:26607837

  3. Structure and Mechanics of Actin Cortex Contained in Vesicles

    NASA Astrophysics Data System (ADS)

    Limozin, Laurent; Roth, Alexander; Sackmann, Erich

    2003-03-01

    We designed giant phospholipid vesicles containing actin filaments as an elementary mechanical cell model. G-actin is polymerized inside the vesicles through ionophore-mediated Mg++ entry and the filaments are bound electrostatically to the membrane through lipids with amino-polyethyleneglycol (PEG) headgroups forming a shell beneath the membrane. The density of this cortex is varied by changing the initial actin concentration. A magnetic micrometric bead attached on the top of a sedimented vesicle is pulled vertically while horizontal and vertical displacements of the bead are simulatenously tracked by microscopy. Linear response allows to determine the bending and shear moduli of the actin-membrane complexe.

  4. Inverse method for simultaneous determination of soil water flux density and thermal properties with a penta-needle heat pulse probe

    NASA Astrophysics Data System (ADS)

    Yang, Changbing; Sakai, Masaru; Jones, Scott B.

    2013-09-01

    An accurate method for determination of in situ soil water flux density continues to be the most sought after and yet elusive hydrologic measurement. The penta-needle heat pulse probe (PHPP) employs a central heater needle surrounded by an orthogonal arrangement of four thermistor needles for two-component water flux density estimation. An analytical solution and inverse fitting method are presented for simultaneous estimation of thermal properties and soil water flux density using PHPP measurements. The approach yields estimates of both components of the flux in a plane normal to the axis of the PHPP needles. The method was evaluated using data measured by PHPPs in a laboratory experiment using a wide range of saturated water fluxes ranging from 1.2 to 33,200 cm d-1. Improved water flux density determination was achieved from zero-flux adjusted estimates of the apparent heater-thermistor radii, radj, which were used in the inverse analysis. Thermal diffusivity and conductivity were estimated with coefficients of variation less than 1.35%, indicating that the inverse problem is well posed and yields unique parameter estimates when water flux is less than 2000 cm d-1. Estimates of the x and y components of water flux density agreed well with measured water fluxes up to 7000 cm d-1 exhibiting R2 values greater than 0.976. Estimation of water flow direction based on 2-D water flux density was in good agreement with installation angle for water fluxes ranging from 10 to 7000 cm d-1.

  5. Actin Automata with Memory

    NASA Astrophysics Data System (ADS)

    Alonso-Sanz, Ramón; Adamatzky, Andy

    Actin is a globular protein which forms long polar filaments in eukaryotic. The actin filaments play the roles of cytoskeleton, motility units, information processing and learning. We model actin filament as a double chain of finite state machines, nodes, which take states “0” and “1”. The states are abstractions of absence and presence of a subthreshold charge on actin units corresponding to the nodes. All nodes update their state in parallel to discrete time. A node updates its current state depending on states of two closest neighbors in the node chain and two closest neighbors in the complementary chain. Previous models of actin automata consider momentary state transitions of nodes. We enrich the actin automata model by assuming that states of nodes depend not only on the current states of neighboring node but also on their past states. Thus, we assess the effect of memory of past states on the dynamics of acting automata. We demonstrate in computational experiments that memory slows down propagation of perturbations, decrease entropy of space-time patterns generated, transforms traveling localizations to stationary oscillators, and stationary oscillations to still patterns.

  6. Demonstartion of density dependence of x-ray flux in a laser-driven hohlraum

    SciTech Connect

    Young, P E; Rosen, M D; Hammer, J H; Hsing, W S; Glendinning, S G; Turner, R E; Kirkwood, R; Schein, J; Sorce, C; Satcher, J; Hamza, A; Reibold, R A; Hibbard, R; Landen, O; Reighard, A; McAlpin, S; Stevenson, M; Thomas, B

    2008-02-11

    Experiments have been conducted using laser-driven cylindrical hohlraums whose walls are machined from Ta{sub 2}O{sub 5} foams of 100 mg/cc and 4 g/cc densities. Measurements of the radiation temperature demonstrate that the lower density walls produce higher radiation temperatures than the high density walls. This is the first experimental demonstration of the prediction that this would occur [M. D. Rosen and J. H. Hammer, Phys. Rev. E 72, 056403 (2005)]. For high density walls, the radiation front propagates subsonically, and part of the absorbed energy is wasted by the flow kinetic energy. For the lower wall density, the front velocity is supersonic and can devote almost all of the absorbed energy to heating the wall.

  7. Capping Protein Modulates the Dynamic Behavior of Actin Filaments in Response to Phosphatidic Acid in Arabidopsis[C][W

    PubMed Central

    Li, Jiejie; Henty-Ridilla, Jessica L.; Huang, Shanjin; Wang, Xia; Blanchoin, Laurent; Staiger, Christopher J.

    2012-01-01

    Remodeling of actin filament arrays in response to biotic and abiotic stimuli is thought to require precise control over the generation and availability of filament ends. Heterodimeric capping protein (CP) is an abundant filament capper, and its activity is inhibited by membrane signaling phospholipids in vitro. How exactly CP modulates the properties of filament ends in cells and whether its activity is coordinated by phospholipids in vivo is not well understood. By observing directly the dynamic behavior of individual filament ends in the cortical array of living Arabidopsis thaliana epidermal cells, we dissected the contribution of CP to actin organization and dynamics in response to the signaling phospholipid, phosphatidic acid (PA). Here, we examined three cp knockdown mutants and found that reduced CP levels resulted in more dynamic activity at filament ends, and this significantly enhanced filament-filament annealing and filament elongation from free ends. The cp mutants also exhibited more dense actin filament arrays. Treatment of wild-type cells with exogenous PA phenocopied the actin-based defects in cp mutants, with an increase in the density of filament arrays and enhanced annealing frequency. These cytoskeletal responses to exogenous PA were completely abrogated in cp mutants. Our data provide compelling genetic evidence that the end-capping activity of CP is inhibited by membrane signaling lipids in eukaryotic cells. Specifically, CP acts as a PA biosensor and key transducer of fluxes in membrane signaling phospholipids into changes in actin cytoskeleton dynamics. PMID:22960908

  8. The Molecular Evolution of Actin

    PubMed Central

    Hightower, Robin C.; Meagher, Richard B.

    1986-01-01

    We have investigated the molecular evolution of plant and nonplant actin genes comparing nucleotide and amino acid sequences of 20 actin genes. Nucleotide changes resulting in amino acid substitutions (replacement substitutions) ranged from 3–7% for all pairwise comparisons of animal actin genes with the following exceptions. Comparisons between higher animal muscle actin gene sequences and comparisons between higher animal cytoplasmic actin gene sequences indicated <3% divergence. Comparisons between plant and nonplant actin genes revealed, with two exceptions, 11–15% replacement substitution. In the analysis of plant actins, replacement substitution between soybean actin genes SAc1, SAc3, SAc4 and maize actin gene MAc1 ranged from 8–10%, whereas these members within the soybean actin gene family ranged from 6–9% replacement substitution. The rate of sequence divergence of plant actin sequences appears to be similar to that observed for animal actins. Furthermore, these and other data suggest that the plant actin gene family is ancient and that the families of soybean and maize actin genes have diverged from a single common ancestral plant actin gene that originated long before the divergence of monocots and dicots. The soybean actin multigene family encodes at least three classes of actin. These classes each contain a pair of actin genes that have been designated kappa (SAc1, SAc6), lambda (SAc2, SAc4) and mu (SAc3, SAc7). The three classes of soybean actin are more divergent in nucleotide sequence from one another than higher animal cytoplasmic actin is divergent from muscle actin. The location and distribution of amino acid changes were compared between actin proteins from all sources. A comparison of the hydropathy of all actin sequences, except from Oxytricha, indicated a strong similarity in hydropathic character between all plant and nonplant actins despite the greater number of replacement substitutions in plant actins. These protein sequence

  9. Observation of the X-ray source Sco X-1 from Skylab. [radiant flux density

    NASA Technical Reports Server (NTRS)

    Wilson, R. M.

    1977-01-01

    An attempt to observe the discrete X-ray source Sco X-1 on 20 September 1973 between 0856 and 0920 UT is reported. Data obtained with the ATM/S-056 X-ray event analyzer, in particular the flux observed with the 1.71 to 4.96 KeV counter, is analyzed. No photographic image of the source was obtained because Sco X-1 was outside the field of view of the X-ray telescope.

  10. Optimization of Magneto-Rheological Damper for Maximizing Magnetic Flux Density in the Fluid Flow Gap Through FEA and GA Approaches

    NASA Astrophysics Data System (ADS)

    Krishna, Hemanth; Kumar, Hemantha; Gangadharan, Kalluvalappil

    2016-06-01

    A magneto rheological (MR) fluid damper offers cost effective solution for semiactive vibration control in an automobile suspension. The performance of MR damper is significantly depends on the electromagnetic circuit incorporated into it. The force developed by MR fluid damper is highly influenced by the magnetic flux density induced in the fluid flow gap. In the present work, optimization of electromagnetic circuit of an MR damper is discussed in order to maximize the magnetic flux density. The optimization procedure was proposed by genetic algorithm and design of experiments techniques. The result shows that the fluid flow gap size less than 1.12 mm cause significant increase of magnetic flux density.

  11. A state-space modeling approach to estimating canopy conductance and associated uncertainties from sap flux density data.

    PubMed

    Bell, David M; Ward, Eric J; Oishi, A Christopher; Oren, Ram; Flikkema, Paul G; Clark, James S

    2015-07-01

    Uncertainties in ecophysiological responses to environment, such as the impact of atmospheric and soil moisture conditions on plant water regulation, limit our ability to estimate key inputs for ecosystem models. Advanced statistical frameworks provide coherent methodologies for relating observed data, such as stem sap flux density, to unobserved processes, such as canopy conductance and transpiration. To address this need, we developed a hierarchical Bayesian State-Space Canopy Conductance (StaCC) model linking canopy conductance and transpiration to tree sap flux density from a 4-year experiment in the North Carolina Piedmont, USA. Our model builds on existing ecophysiological knowledge, but explicitly incorporates uncertainty in canopy conductance, internal tree hydraulics and observation error to improve estimation of canopy conductance responses to atmospheric drought (i.e., vapor pressure deficit), soil drought (i.e., soil moisture) and above canopy light. Our statistical framework not only predicted sap flux observations well, but it also allowed us to simultaneously gap-fill missing data as we made inference on canopy processes, marking a substantial advance over traditional methods. The predicted and observed sap flux data were highly correlated (mean sensor-level Pearson correlation coefficient = 0.88). Variations in canopy conductance and transpiration associated with environmental variation across days to years were many times greater than the variation associated with model uncertainties. Because some variables, such as vapor pressure deficit and soil moisture, were correlated at the scale of days to weeks, canopy conductance responses to individual environmental variables were difficult to interpret in isolation. Still, our results highlight the importance of accounting for uncertainty in models of ecophysiological and ecosystem function where the process of interest, canopy conductance in this case, is not observed directly. The StaCC modeling

  12. Spatial variations in xylem sap flux density in the trunk of orchard-grown, mature mango trees under changing soil water conditions.

    PubMed

    Lu, Ping; Müller, Warren J.; Chacko, Elias K.

    2000-05-01

    Circumferential and radial variations in xylem sap flux density in trunks of 13-year-old mango (Mangifera indica L.) trees were investigated with Granier sap flow sensor probes under limiting and non-limiting soil water conditions. Under non-limiting soil water conditions, circumferential variation was substantial, but there was no apparent relationship between sap flux density and aspect (i.e., the radial position of the sensor probes on the trunk relative to the compass). Hourly sap flux densities over 24 hours at different aspects were highly pair-wise correlated. The relationships between different aspects were constant during well-watered periods but highly variable under changing soil water conditions. Sap flux density showed marked radial variation within the trunk and a substantial flux was observed at the center of the trunk. For each selected aspect on each tree, changes in sap flux densities over time at different depths were closely correlated, so flux at a particular depth could be extrapolated as a multiple of flux from 0 to 2 cm beneath the cambium. However, depth profiles of sap flux density differed between trees and even between aspects within a tree, and also varied in an unpredictable manner as soil water conditions changed. Nevertheless, over a period of non-limiting soil water conditions, depth profiles remained relatively constant. Based on the depth profiles obtained during these periods, a method is described for calculating total sap flow in a mango tree from sap flux density at 0-2 cm beneath the cambium. Total daily sap flows obtained were consistent with water use estimated from soil water balance. PMID:12651518

  13. An improved multiphase lattice Boltzmann flux solver for three-dimensional flows with large density ratio and high Reynolds number

    NASA Astrophysics Data System (ADS)

    Wang, Y.; Shu, C.; Yang, L. M.

    2015-12-01

    An improved multiphase lattice Boltzmann flux solver (MLBFS) is proposed in this work for effective simulation of three-dimensional (3D) multiphase flows with large density ratio and high Reynolds number. As a finite volume scheme, the MLBFS originally proposed in [27] applies the finite volume method to solve for macroscopic flow variables directly. The fluxes are reconstructed locally at each cell interface by using the standard LBM solutions. Due to the modeling error of the standard LBM, the reconstructed fluxes deviate from those in the Navier-Stokes equations; and to compensate this error, a complex tensor is introduced in the original MLBFS. However, the computation of the tensor introduces additional complexity and usually needs a relatively thicker interface thickness to maintain numerical stability, which makes the solver be complex and inefficient in the 3D case. To remove this drawback, in this work, a theoretical analysis to the formulations obtained from the Chapman-Enskog expansion is conducted. It is shown that the modeling error can be effectively removed by modifying the computation of the equilibrium density distribution function. With this improvement, the proposed 3D MLBFS not only avoids the calculation of the compensation tensor but also is able to maintain numerical stability with very thin interface thickness. Several benchmark cases, including the challenging droplet impacting on a dry surface, head-on collisions of binary droplets and droplet splashing on a thin film with density ratio 1000 and Reynolds number up to 3000, are studied to validate the proposed solver. The obtained results agree well with the published data.

  14. Lateral Membrane Diffusion Modulated by a Minimal Actin Cortex

    PubMed Central

    Heinemann, Fabian; Vogel, Sven K.; Schwille, Petra

    2013-01-01

    Diffusion of lipids and proteins within the cell membrane is essential for numerous membrane-dependent processes including signaling and molecular interactions. It is assumed that the membrane-associated cytoskeleton modulates lateral diffusion. Here, we use a minimal actin cortex to directly study proposed effects of an actin meshwork on the diffusion in a well-defined system. The lateral diffusion of a lipid and a protein probe at varying densities of membrane-bound actin was characterized by fluorescence correlation spectroscopy (FCS). A clear correlation of actin density and reduction in mobility was observed for both the lipid and the protein probe. At high actin densities, the effect on the protein probe was ∼3.5-fold stronger compared to the lipid. Moreover, addition of myosin filaments, which contract the actin mesh, allowed switching between fast and slow diffusion in the minimal system. Spot variation FCS was in accordance with a model of fast microscopic diffusion and slower macroscopic diffusion. Complementing Monte Carlo simulations support the analysis of the experimental FCS data. Our results suggest a stronger interaction of the actin mesh with the larger protein probe compared to the lipid. This might point toward a mechanism where cortical actin controls membrane diffusion in a strong size-dependent manner. PMID:23561523

  15. FeCo-Zr-O nanogranular soft-magnetic thin films with a high magnetic flux density

    NASA Astrophysics Data System (ADS)

    Ohnuma, S.; Fujimori, H.; Masumoto, T.; Xiong, X. Y.; Ping, D. H.; Hono, K.

    2003-02-01

    Soft-magnetic thin films with high magnetic flux densities of about 23 kG have been fabricated in the (Fe-Co)-Zr-O nanogranular system. The films were prepared by reactive sputtering under an oxygen-argon atmosphere using a target of Fe-Co-Zr alloys. The microstructure was composed of base-centered-cubic Fe-Co nanograins, where nanoparticles of amorphous Zr oxide are dispersed. These Zr-oxide nanoparticles are thought to hinder the growth of Fe-Co grains during the film deposition, causing low coercivity.

  16. Nuclear and cytoplasmic actin in dinoflagellates.

    PubMed

    Soyer-Gobillard, M O; Ausseil, J; Géraud, M L

    1996-01-01

    Experiments using monoclonal and polyclonal anti-actin antibodies allowed us to demonstrate the presence of F- or G-actin in original protists, dinoflagellates, either by biochemistry, immunofluorescence and in TEM. SDS-PAGE electrophoresis and immunoblottings made either from total or nuclear protein extracts revealed the presence of a 44-kDa band reacting with monoclonal anti-actin antibody in two species, Prorocentrum micans and Crypthecodinium cohnii, and thus demonstrated the presence of actin in nuclear and cytoplasmic fractions. After squash preparation of P micans cells, actin was identified within the nucleus and in some regions of the cytoplasm by immunofluorescence microscopy. Labelling of both the nucleolus and the centrosome region was evident together with amorphous nucleoplasmic material surrounding the chromosomes. The use of cryosections of intact P micans and C cohnii cells for immunofluorescence along with staining with DAPI to delineate the chromosomes themselves, yielded finer resolution of the intranuclear network labelling pattern and allowed us to complete our observations, in particular on the cytoplasmic labelling. In P micans, in addition to the centrosome region, the cytoplasmic channels passing through the nucleus in dividing cells are labelled. In C cohnii, the cortex, the centrosome region, the cytoplasmic channels, the region surrounding the nucleus, the filaments linking it to the cortex and the cleavage furrow are also labelled. In the nucleus of the two species, there is a prominent "weft' of fine actin filaments in the nucleoplasm forming a matrix of varying density around the persistent chromosomes. This actin matrix, of unknown function, is most conspicuous at the end of the S-phase of the cell cycle. Fluorescent derivatives of phalloidin, used as diagnostic cytochemical probes for polymeric actin (F-actin), gave similar results. Positive TEM immunolabelling of intranuclear actin confirms its presence in the nucleoplasm, in the

  17. Intranuclear Actin Regulates Osteogenesis

    PubMed Central

    Sen, Buer; Xie, Zhihui; Uzer, Gunes; Thompson, William R.; Styner, Maya; Wu, Xin; Rubin, Janet

    2016-01-01

    Depolymerization of the actin cytoskeleton induces nuclear trafficking of regulatory proteins and global effects on gene transcription. We here show that in mesenchymal stem cells (MSCs), cytochalasin D treatment causes rapid cofilin-/importin-9-dependent transfer of G-actin into the nucleus. The continued presence of intranuclear actin, which forms rod-like structures that stain with phalloidin, is associated with induction of robust expression of the osteogenic genes osterix and osteocalcin in a Runx2-dependent manner, and leads to acquisition of osteogenic phenotype. Adipogenic differentiation also occurs, but to a lesser degree. Intranuclear actin leads to nuclear export of Yes-associated protein (YAP); maintenance of nuclear YAP inhibits Runx2 initiation of osteogenesis. Injection of cytochalasin into the tibial marrow space of live mice results in abundant bone formation within the space of 1 week. In sum, increased intranuclear actin forces MSC into osteogenic lineage through controlling Runx2 activity; this process may be useful for clinical objectives of forming bone. PMID:26140478

  18. Simulation study of geometric shape factor approach to estimating earth emitted flux densities from wide field-of-view radiation measurements

    NASA Technical Reports Server (NTRS)

    Weaver, W. L.; Green, R. N.

    1980-01-01

    A study was performed on the use of geometric shape factors to estimate earth-emitted flux densities from radiation measurements with wide field-of-view flat-plate radiometers on satellites. Sets of simulated irradiance measurements were computed for unrestricted and restricted field-of-view detectors. In these simulations, the earth radiation field was modeled using data from Nimbus 2 and 3. Geometric shape factors were derived and applied to these data to estimate flux densities on global and zonal scales. For measurements at a satellite altitude of 600 km, estimates of zonal flux density were in error 1.0 to 1.2%, and global flux density errors were less than 0.2%. Estimates with unrestricted field-of-view detectors were about the same for Lambertian and non-Lambertian radiation models, but were affected by satellite altitude. The opposite was found for the restricted field-of-view detectors.

  19. Structure of the F-actin-tropomyosin complex.

    PubMed

    von der Ecken, Julian; Müller, Mirco; Lehman, William; Manstein, Dietmar J; Penczek, Pawel A; Raunser, Stefan

    2015-03-01

    Filamentous actin (F-actin) is the major protein of muscle thin filaments, and actin microfilaments are the main component of the eukaryotic cytoskeleton. Mutations in different actin isoforms lead to early-onset autosomal dominant non-syndromic hearing loss, familial thoracic aortic aneurysms and dissections, and multiple variations of myopathies. In striated muscle fibres, the binding of myosin motors to actin filaments is mainly regulated by tropomyosin and troponin. Tropomyosin also binds to F-actin in smooth muscle and in non-muscle cells and stabilizes and regulates the filaments there in the absence of troponin. Although crystal structures for monomeric actin (G-actin) are available, a high-resolution structure of F-actin is still missing, hampering our understanding of how disease-causing mutations affect the function of thin muscle filaments and microfilaments. Here we report the three-dimensional structure of F-actin at a resolution of 3.7 Å in complex with tropomyosin at a resolution of 6.5 Å, determined by electron cryomicroscopy. The structure reveals that the D-loop is ordered and acts as a central region for hydrophobic and electrostatic interactions that stabilize the F-actin filament. We clearly identify map density corresponding to ADP and Mg(2+) and explain the possible effect of prominent disease-causing mutants. A comparison of F-actin with G-actin reveals the conformational changes during filament formation and identifies the D-loop as their key mediator. We also confirm that negatively charged tropomyosin interacts with a positively charged groove on F-actin. Comparison of the position of tropomyosin in F-actin-tropomyosin with its position in our previously determined F-actin-tropomyosin-myosin structure reveals a myosin-induced transition of tropomyosin. Our results allow us to understand the role of individual mutations in the genesis of actin- and tropomyosin-related diseases and will serve as a strong foundation for the targeted

  20. A Dual-Mode Actinic EUV Mask Inspection Tool

    SciTech Connect

    Liu, Y; Barty, A; Gullikson, E; S.Taylor, J; Liddle, J A; Wood, O

    2005-03-21

    To qualify the performance of non-actinic inspection tools, a novel EUV mask inspection system has been installed at the Advanced Light Source (ALS) synchrotron facility at Lawrence Berkeley National Laboratory. Similar to the older generation actinic mask inspection tool, the new system can operate in scanning mode, when mask blanks are scanned for defects using 13.5-nm in-band radiation to identify and map all locations on the mask that scatter a significant amount of EUV light. By modifying and optimizing beamline optics (11.3.2 at ALS) and replacing K-B focusing mirrors with a high quality Schwarzschild illuminator, the new system achieves an order of magnitude improvement on in-band EUV flux density at the mask, enabling faster scanning speed and higher sensitivity to smaller defects. Moreover, the system can also operate in imaging mode, when it becomes a zone-plate-based full-field EUV microscope with spatial resolution better than 100 nm. The microscope utilizes an off-axis setup, making it possible to obtain bright field images over a field-of-view of 5 x 5 {micro}m.

  1. Myosins, Actin and Autophagy.

    PubMed

    Kruppa, Antonina J; Kendrick-Jones, John; Buss, Folma

    2016-08-01

    Myosin motor proteins working together with the actin cytoskeleton drive a wide range of cellular processes. In this review, we focus on their roles in autophagy - the pathway the cell uses to ensure homeostasis by targeting pathogens, misfolded proteins and damaged organelles for degradation. The actin cytoskeleton regulated by a host of nucleating, anchoring and stabilizing proteins provides the filament network for the delivery of essential membrane vesicles from different cellular compartments to the autophagosome. Actin networks have also been implicated in structurally supporting the expanding phagophore, moving autophagosomes and enabling efficient fusion with the lysosome. Only a few myosins have so far been shown to play a role in autophagy. Non-muscle myosin IIA functions in the early stages delivering membrane for the initial formation of the autophagosome, whereas myosin IC and myosin VI are involved in the final stages providing specific membranes for autophagosome maturation and its fusion with the lysosome. PMID:27146966

  2. BOREAS RSS-17 Xylem Flux Density Measurements at the SSA-OBS Site

    NASA Technical Reports Server (NTRS)

    Zimmerman, Reiner; Way, JoBea; McDonald, Kyle; Nickeson, Jaime (Editor); Hall, Forrest G. (Editor); Smith, David E. (Technical Monitor)

    2000-01-01

    As part of its efforts to determine environmental and phenological states from radar imagery, the Boreal Ecosystem-Atmosphere Study (BOREAS) Remote Sensing Science (RSS)-17 team collected in situ tree xylem flow measurements for one growing season on five Picea mariana (black spruce) trees. The data were collected to obtain information on the temporal and spatial variability in water uptake by trees in the Southern Study Area-Old Black Spruce (SSA-OBS) stand in the BOREAS SSA. Temporally, the data were collected in 30-minute intervals for 120 days from 31 May 1994 until 27 September 1994. The data are stored in tabular ASCII files. The xylem flux data are available from the Earth Observing System Data and Information System (EOSDIS) Oak Ridge National Laboratory (ORNL) Distributed Active Archive Center (DAAC). The data files are available on a CD-ROM (see document number 20010000884).

  3. Emergence flux declines disproportionately to larval density along a stream metals gradient

    USGS Publications Warehouse

    Schmidt, Travis S.; Kraus, Johanna M.; Walters, David M.; Wanty, Richard B.

    2013-01-01

    Effects of contaminants on adult aquatic insect emergence are less well understood than effects on insect larvae. We compared responses of larval density and adult emergence along a metal contamination gradient. Nonlinear threshold responses were generally observed for larvae and emergers. Larval densities decreased significantly at low metal concentrations but precipitously at concentrations of metal mixtures above aquatic life criteria (Cumulative Criterion Accumulation Ratio (CCAR) ≥ 1). In contrast, adult emergence declined precipitously at low metal concentrations (CCAR ≤ 1), followed by a modest decline above this threshold. Adult emergence was a more sensitive indicator of the effect of low metals concentrations on aquatic insect communities compared to larvae, presumably because emergence is limited by a combination of larval survival and other factors limiting successful emergence. Thus effects of exposure to larvae are not manifest until later in life (during metamorphosis and emergence). This loss in emergence reduces prey subsidies to riparian communities at concentrations considered safe for aquatic life. Our results also challenge the widely held assumption that adult emergence is a constant proportion of larval densities in all streams.

  4. A Mechanism for Actin Filament Severing by Malaria Parasite Actin Depolymerizing Factor 1 via a Low Affinity Binding Interface*

    PubMed Central

    Wong, Wilson; Webb, Andrew I.; Olshina, Maya A.; Infusini, Giuseppe; Tan, Yan Hong; Hanssen, Eric; Catimel, Bruno; Suarez, Cristian; Condron, Melanie; Angrisano, Fiona; NebI, Thomas; Kovar, David R.; Baum, Jake

    2014-01-01

    Actin depolymerizing factor (ADF)/cofilins are essential regulators of actin turnover in eukaryotic cells. These multifunctional proteins facilitate both stabilization and severing of filamentous (F)-actin in a concentration-dependent manner. At high concentrations ADF/cofilins bind stably to F-actin longitudinally between two adjacent actin protomers forming what is called a decorative interaction. Low densities of ADF/cofilins, in contrast, result in the optimal severing of the filament. To date, how these two contrasting modalities are achieved by the same protein remains uncertain. Here, we define the proximate amino acids between the actin filament and the malaria parasite ADF/cofilin, PfADF1 from Plasmodium falciparum. PfADF1 is unique among ADF/cofilins in being able to sever F-actin but do so without stable filament binding. Using chemical cross-linking and mass spectrometry (XL-MS) combined with structure reconstruction we describe a previously overlooked binding interface on the actin filament targeted by PfADF1. This site is distinct from the known binding site that defines decoration. Furthermore, total internal reflection fluorescence (TIRF) microscopy imaging of single actin filaments confirms that this novel low affinity site is required for F-actin severing. Exploring beyond malaria parasites, selective blocking of the decoration site with human cofilin (HsCOF1) using cytochalasin D increases its severing rate. HsCOF1 may therefore also use a decoration-independent site for filament severing. Thus our data suggest that a second, low affinity actin-binding site may be universally used by ADF/cofilins for actin filament severing. PMID:24371134

  5. Topside equatorial ionospheric density, temperature, and composition under equinox, low solar flux conditions

    NASA Astrophysics Data System (ADS)

    Hysell, D. L.; Milla, M. A.; Rodrigues, F. S.; Varney, R. H.; Huba, J. D.

    2015-05-01

    We present observations of the topside ionosphere made at the Jicamarca Radio Observatory in March and September 2013, made using a full-profile analysis approach. Recent updates to the methodology employed at Jicamarca are also described. Measurements of plasma number density, electron and ion temperatures, and hydrogen and helium ion fractions up to 1500 km altitude are presented for 3 days in March and September. The main features of the observations include a sawtooth-like diurnal variation in ht, the transition height where the O+ ion fraction falls to 50%, the appearance of weak He+ layers just below ht, and a dramatic increase in plasma temperature at dawn followed by a sharp temperature depression around local noon. These features are consistent from day to day and between March and September. Coupled Ion Neutral Dynamics Investigation data from the Communication Navigation Outage Forecast System satellite are used to help validate the March Jicamarca data. The SAMI2-PE model was able to recover many of the features of the topside observations, including the morphology of the plasma density profiles and the light-ion composition. The model, forced using convection speeds and meridional thermospheric winds based on climatological averages, did not reproduce the extreme temperature changes in the topside between sunrise and noon. Some possible causes of the discrepancies are discussed.

  6. Secure Mass Measurements from Transit Timing: 10 Kepler Exoplanets between 3 and 8 M⊕ with Diverse Densities and Incident Fluxes

    NASA Astrophysics Data System (ADS)

    Jontof-Hutter, Daniel; Ford, Eric B.; Rowe, Jason F.; Lissauer, Jack J.; Fabrycky, Daniel C.; Van Laerhoven, Christa; Agol, Eric; Deck, Katherine M.; Holczer, Tomer; Mazeh, Tsevi

    2016-03-01

    We infer dynamical masses in eight multiplanet systems using transit times measured from Kepler's complete data set, including short-cadence data where available. Of the 18 dynamical masses that we infer, 10 pass multiple tests for robustness. These are in systems Kepler-26 (KOI-250), Kepler-29 (KOI-738), Kepler-60 (KOI-2086), Kepler-105 (KOI-115), and Kepler-307 (KOI-1576). Kepler-105 c has a radius of 1.3 R⊕ and a density consistent with an Earth-like composition. Strong transit timing variation (TTV) signals were detected from additional planets, but their inferred masses were sensitive to outliers or consistent solutions could not be found with independently measured transit times, including planets orbiting Kepler-49 (KOI-248), Kepler-57 (KOI-1270), Kepler-105 (KOI-115), and Kepler-177 (KOI-523). Nonetheless, strong upper limits on the mass of Kepler-177 c imply an extremely low density of ˜0.1 g cm-3. In most cases, individual orbital eccentricities were poorly constrained owing to degeneracies in TTV inversion. For five planet pairs in our sample, strong secular interactions imply a moderate to high likelihood of apsidal alignment over a wide range of possible eccentricities. We also find solutions for the three planets known to orbit Kepler-60 in a Laplace-like resonance chain. However, nonlibrating solutions also match the transit timing data. For six systems, we calculate more precise stellar parameters than previously known, enabling useful constraints on planetary densities where we have secure mass measurements. Placing these exoplanets on the mass-radius diagram, we find that a wide range of densities is observed among sub-Neptune-mass planets and that the range in observed densities is anticorrelated with incident flux.

  7. Phototactic number-density flux in the localized bioconvection of Euglena gracilis

    NASA Astrophysics Data System (ADS)

    Shoji, Erika; Suematsu, Nobuhiko; Nishimori, Hiraku; Awazu, Akinori; Izumi, Shunsuke; Iima, Makoto

    2014-11-01

    Euglena gracilis is a unicellular phototactic flagellate; it escapes from light sources if the light intensity is higher than 200 W/m2 (negative phototaxis). When the suspension of E. gracilis is illuminated from the bottom by strong light, bioconvection patterns are generated. In the case of E. gracilis, the patterns can be spatially localized. The localization mechanism has not been clarified. We report experimental results related to the localization mechanism. In particular, we experimentally measured the strength of the phototaxis in the lateral direction as well as vertical direction. We prepared a thin container in which the suspension is included, and gave the linearly-changing light intensity. We found the number density gets a peak at a particular light intensity, which never happens if the suspension has the vertical phototaxis only. Further, we succeeded in getting the function representing lateral phototaxis. The relationship between the measured functions and the localized convection cells will be also reported.

  8. Curved trajectories of actin-based motility in two dimensions

    NASA Astrophysics Data System (ADS)

    Wen, Fu-Lai; Leung, Kwan-tai; Chen, Hsuan-Yi

    2012-05-01

    Recent experiments have reported fascinating geometrical trajectories for actin-based motility of bacteria Listeria monocytogenes and functionalized beads. To understand the physical mechanism for these trajectories, we constructed a phenomenological model to study the motion of an actin-propelled disk in two dimensions. In our model, the force and actin density on the surface of the disk are influenced by the translation and rotation of the disk, which in turn is induced by the asymmetric distributions of those densities. We show that this feedback can destabilize a straight trajectory, leading to circular, S-shape and other geometrical trajectories observed in the experiments through bifurcations in the distributions of the force and actin density. The relation between our model and the models for self-propelled deformable particles is emphasized and discussed.

  9. Wavelet and Fractal Analysis of Remotely Sensed Surface Temperature with Applications to Estimation of Surface Sensible Heat Flux Density

    NASA Technical Reports Server (NTRS)

    Schieldge, John

    2000-01-01

    Wavelet and fractal analyses have been used successfully to analyze one-dimensional data sets such as time series of financial, physical, and biological parameters. These techniques have been applied to two-dimensional problems in some instances, including the analysis of remote sensing imagery. In this respect, these techniques have not been widely used by the remote sensing community, and their overall capabilities as analytical tools for use on satellite and aircraft data sets is not well known. Wavelet and fractal analyses have the potential to provide fresh insight into the characterization of surface properties such as temperature and emissivity distributions, and surface processes such as the heat and water vapor exchange between the surface and the lower atmosphere. In particular, the variation of sensible heat flux density as a function of the change In scale of surface properties Is difficult to estimate, but - in general - wavelets and fractals have proved useful in determining the way a parameter varies with changes in scale. We present the results of a limited study on the relationship between spatial variations in surface temperature distribution and sensible heat flux distribution as determined by separate wavelet and fractal analyses. We analyzed aircraft imagery obtained in the thermal infrared (IR) bands from the multispectral TIMS and hyperspectral MASTER airborne sensors. The thermal IR data allows us to estimate the surface kinetic temperature distribution for a number of sites in the Midwestern and Southwestern United States (viz., San Pedro River Basin, Arizona; El Reno, Oklahoma; Jornada, New Mexico). The ground spatial resolution of the aircraft data varied from 5 to 15 meters. All sites were instrumented with meteorological and hydrological equipment including surface layer flux measuring stations such as Bowen Ratio systems and sonic anemometers. The ground and aircraft data sets provided the inputs for the wavelet and fractal analyses

  10. GaN Etch Rates Compared with Atomic Chlorine Density and Ion Flux in an Argon/Chlorine Inductively Coupled Plasma

    NASA Astrophysics Data System (ADS)

    Mahony, C. M. O.; Rizvi, S. A.; Maguire, P. D.; Garcia, F.; Graham, W. G.

    2004-09-01

    We present GaN etch rates (maximum 700nm/min), atomic chlorine densities (via Laser Induced Fluorescence at 200W RF power), positive ion densities (Langmuir probe) and positive ion wall flux (capacitive planar probe) using an Inductively Coupled Plasma as a function of chlorine in argon gas fraction from 0% to 100% at maximum RF power and pressure of 400 W and 20 mTorr respectively. In general, with chlorine addition, etch rates rise initially then tend to saturate at fractions above 50% Cl_2. Wall flux and n^+ approximate the inverse of this behaviour. The atomic chlorine density at 200W RF power rises monotonically with a pronounced inflection near 50% Cl_2. The positive ion wall flux - atomic chlorine density product strongly correlates with etch rate suggesting physical etching dominates below 50% Cl2 and chemical processes above. This is reflected in changes of the Ga/N surface stoichiometry, determined by XPS analysis.

  11. Scanning micro-Hall probe mapping of magnetic flux distributions and current densities in YBa2Cu3O7 thin films

    NASA Technical Reports Server (NTRS)

    Xing, W.; Heinrich, B.; Zhou, HU; Fife, A. A.; Cragg, A. R.; Grant, P. D.

    1995-01-01

    Mapping of the magnetic flux density B(sub z) (perpendicular to the film plane) for a YBa2Cu3O7 thin-film sample was carried out using a scanning micro-Hall probe. The sheet magnetization and sheet current densities were calculated from the B(sub z) distributions. From the known sheet magnetization, the tangential (B(sub x,y)) and normal components of the flux density B were calculated in the vicinity of the film. It was found that the sheet current density was mostly determined by 2B(sub x,y)/d, where d is the film thickness. The evolution of flux penetration as a function of applied field will be shown.

  12. Scanning micro-Hall probe mapping of magnetic flux distributions and current densities in YBa{sub 2}Cu{sub 3}O{sub 7}

    SciTech Connect

    Xing, W.; Heinrich, B.; Zhou, H.

    1994-12-31

    Mapping of the magnetic flux density B{sub z} (perpendicular to the film plane) for a YBa{sub 2}Cu{sub 3}O{sub 7} thin-film sample was carried out using a scanning micro-Hall probe. The sheet magnetization and sheet current densities were calculated from the B{sub z} distributions. From the known sheet magnetization, the tangential (B{sub x,y}) and normal components of the flux density B were calculated in the vicinity of the film. It was found that the sheet current density was mostly determined by 2B{sub x,y}/d, where d is the film thickness. The evolution of flux penetration as a function of applied field will be shown.

  13. Actin dynamics and the evolution of the memory trace.

    PubMed

    Rudy, Jerry W

    2015-09-24

    The goal of this essay is to link the regulation of actin dynamics to the idea that the synaptic changes that support long-term potentiation and memory evolve in temporally overlapping stages-generation, stabilization, and consolidation. Different cellular/molecular processes operate at each stage to change the spine cytoarchitecture and, in doing so, alter its function. Calcium-dependent processes that degrade the actin cytoskeleton network promote a rapid insertion of AMPA receptors into the post synaptic density, which increases a spine's capacity to express a potentiated response to glutamate. Other post-translation events then begin to stabilize and expand the actin cytoskeleton by increasing the filament actin content of the spine and reorganizing it to be resistant to depolymerizing events. Disrupting actin polymerization during this stabilization period is a terminal event-the actin cytoskeleton shrinks and potentiated synapses de-potentiate and memories are lost. Late-arriving, new proteins may consolidate changes in the actin cytoskeleton. However, to do so requires a stabilized actin cytoskeleton. The now enlarged spine has properties that enable it to capture other newly transcribed mRNAs or their protein products and thus enable the synaptic changes that support LTP and memory to be consolidated and maintained. This article is part of a Special Issue entitled SI: Brain and Memory. PMID:25498985

  14. Comparison of Uniform and Non-uniform Water Flux Density Approaches Applied on a Mathematical Model of Heat Transfer and Solidification for a Continuous Casting of Round Billets

    NASA Astrophysics Data System (ADS)

    Assuncao, Charles Sostenes; Tavares, Roberto Parreiras; Oliveira, Guilherme; Pereira, Luiz Carlos

    2015-02-01

    In the present work, the water flux densities of nozzles with flat jet and full cone jet were experimentally measured using an apparatus in industrial scale that reproduces the secondary cooling of the continuous casting of round billets of Vallourec Tubos do Brasil. A mathematical function was defined to express the water flux density in both longitudinal and angular directions of the strand. A mathematical model for heat transfer and solidification for the continuous casting of round billets was developed applying the experimental water flux density profile, establishing a non-uniform water distribution approach. The mathematical model was validated by experimental measurements of the billet superficial temperature, performed at the industrial plant. The results of the mathematical model using both uniform and non-uniform water flux density approaches were compared. The non-uniform water distribution approach enabled to identify important variations of the heat transfer coefficients and the billet temperatures, especially in the first cooling zones where the steel temperature is higher, and to assess more accurately the local effects of the water distribution on the thermal behavior of the strand. The non-uniform water flux density approach applied to the mathematical model was a useful and more accurate tool to improve the comprehension of the thermal behavior of the steel along the secondary cooling.

  15. Symmetry breaking in actin gels - Implications for cellular motility

    NASA Astrophysics Data System (ADS)

    John, Karin; Peyla, Philippe; Misbah, Chaouqi

    2007-03-01

    The physical origin of cell motility is not fully understood. Recently minimal model systems have shown, that polymerizing actin itself can produce a motile force, without the help of motor proteins. Pathogens like Shigella or Listeria use actin to propel themselves forward in their host cell. The same process can be mimicked with polystyrene beads covered with the activating protein ActA, which reside in a solution containing actin monomers. ActA induces the growth of an actin gel at the bead surface. Initially the gel grows symmetrically around the bead until a critical size is reached. Subsequently one observes a symmetry breaking and the gel starts to grow asymmetrically around the bead developing a tail of actin at one side. This symmetry breaking is accompanied by a directed movement of the bead, with the actin tail trailing behind the bead. Force generation relies on the combination of two properties: growth and elasticity of the actin gel. We study this phenomenon theoretically within the framework of a linear elasticity theory and linear flux-force relationships for the evolution of an elastic gel around a hard sphere. Conditions for a parity symmetry breaking are identified analytically and illustrated numerically with the help of a phasefield model.

  16. Actin Dynamics in Growth Cone Motility and Navigation

    PubMed Central

    Gomez, Timothy M.; Letourneau, Paul C.

    2014-01-01

    Motile growth cones lead growing axons through developing tissues to synaptic targets. These behaviors depend on the organization and dynamics of actin filaments that fill the growth cone leading margin (peripheral (P-) domain). Actin filament organization in growth cones is regulated by actin-binding proteins that control all aspects of filament assembly, turnover, interactions with other filaments and cytoplasmic components, and participation in producing mechanical forces. Actin filament polymerization drives protrusion of sensory filopodia and lamellipodia, and actin filament connections to the plasma membrane link the filament network to adhesive contacts of filopodia and lamellipodia with other surfaces. These contacts stabilize protrusions and transduce mechanical forces generated by actomyosin activity into traction that pulls an elongating axon along the path towards its target. Adhesive ligands and extrinsic guidance cues bind growth cone receptors and trigger signaling activities involving Rho GTPases, kinases, phosphatases, cyclic nucleotides and [Ca++] fluxes. These signals regulate actin binding proteins to locally modulate actin polymerization, interactions and force transduction to steer the growth cone leading margin towards the sources of attractive cues and away from repellent guidance cues. PMID:24164353

  17. Arp2/3 complex-dependent actin networks constrain myosin II function in driving retrograde actin flow.

    PubMed

    Yang, Qing; Zhang, Xiao-Feng; Pollard, Thomas D; Forscher, Paul

    2012-06-25

    The Arp2/3 complex nucleates actin filaments to generate networks at the leading edge of motile cells. Nonmuscle myosin II produces contractile forces involved in driving actin network translocation. We inhibited the Arp2/3 complex and/or myosin II with small molecules to investigate their respective functions in neuronal growth cone actin dynamics. Inhibition of the Arp2/3 complex with CK666 reduced barbed end actin assembly site density at the leading edge, disrupted actin veils, and resulted in veil retraction. Strikingly, retrograde actin flow rates increased with Arp2/3 complex inhibition; however, when myosin II activity was blocked, Arp2/3 complex inhibition now resulted in slowing of retrograde actin flow and veils no longer retracted. Retrograde flow rate increases induced by Arp2/3 complex inhibition were independent of Rho kinase activity. These results provide evidence that, although the Arp2/3 complex and myosin II are spatially segregated, actin networks assembled by the Arp2/3 complex can restrict myosin II-dependent contractility with consequent effects on growth cone motility. PMID:22711700

  18. Compact and high-particle-flux thermal-lithium-beam probe system for measurement of two-dimensional electron density profile

    SciTech Connect

    Shibata, Y. Manabe, T.; Ohno, N.; Takagi, M.; Kajita, S.; Tsuchiya, H.; Morisaki, T.

    2014-09-15

    A compact and high-particle-flux thermal-lithium-beam source for two-dimensional measurement of electron density profiles has been developed. The thermal-lithium-beam oven is heated by a carbon heater. In this system, the maximum particle flux of the thermal lithium beam was ∼4 × 10{sup 19} m{sup −2} s{sup −1} when the temperature of the thermal-lithium-beam oven was 900 K. The electron density profile was evaluated in the small tokamak device HYBTOK-II. The electron density profile was reconstructed using the thermal-lithium-beam probe data and this profile was consistent with the electron density profile measured with a Langmuir electrostatic probe. We confirm that the developed thermal-lithium-beam probe can be used to measure the two-dimensional electron density profile with high time and spatial resolutions.

  19. Simulations of the Cleft Ion Fountain outflows resulting from the passage of Storm Enhanced Density (SED) plasma flux tubes through the dayside cleft auroral processes region

    NASA Astrophysics Data System (ADS)

    Horwitz, James; Zeng, Wen

    2007-10-01

    Foster et al. [2002] reported elevated ionospheric density regions convected from subauroral plasmaspheric regions toward noon, in association with convection of plasmaspheric tails. These Storm Enhanced Density (SED) regions could supply cleft ion fountain outflows. Here, we will utilize our Dynamic Fluid Kinetic (DyFK) model to simulate the entry of a high-density ``plasmasphere-like'' flux tube entering the cleft region and subjected to an episode of wave-driven transverse ion heating. It is found that the O^+ ion density at higher altitudes increases and the density at lower altitudes decreases, following this heating episode, indicating increased fluxes of O^+ ions from the ionospheric source gain sufficient energy to reach higher altitudes after the effects of transverse wave heating. Foster, J. C., P. J. Erickson, A. J. Coster, J. Goldstein, and F. J. Rich, Ionospheric signatures of plasmaspheric tails, Geophys. Res. Lett., 29(13), 1623, doi:10.1029/2002GL015067, 2002.

  20. Solar flux-density distribution due to partially shaded/blocked mirrors using the separation of variables/superposition technique with polynomial and Gaussian sunshapes

    SciTech Connect

    Elsayed, M.; Fathalah, K.A.

    1996-05-01

    In a previous work, the separation of a variable/superposition technique was used to predict the flux density distribution on the receiver surfaces of solar central receiver plants. In this paper further developments of the technique are given. A numerical technique is derived to carry out the convolution of the sunshape and error density functions. Also, a simplified numerical procedure is presented to determine the basic flux density function on which the technique depends. The technique is used to predict the receiver solar flux distribution using two sunshapes, polynomial and Gaussian distributions. The results predicted with the technique are validated by comparison with experimental results from mirrors both with and without partial shading/blocking of their surfaces.

  1. Villin Severing Activity Enhances Actin-based Motility In Vivo

    PubMed Central

    Revenu, Céline; Courtois, Matthieu; Michelot, Alphée; Sykes, Cécile; Louvard, Daniel

    2007-01-01

    Villin, an actin-binding protein associated with the actin bundles that support microvilli, bundles, caps, nucleates, and severs actin in a calcium-dependant manner in vitro. We hypothesized that the severing activity of villin is responsible for its reported role in enhancing cell plasticity and motility. To test this hypothesis, we chose a loss of function strategy and introduced mutations in villin based on sequence comparison with CapG. By pyrene-actin assays, we demonstrate that this mutant has a strongly reduced severing activity, whereas nucleation and capping remain unaffected. The bundling activity and the morphogenic effects of villin in cells are also preserved in this mutant. We thus succeeded in dissociating the severing from the three other activities of villin. The contribution of villin severing to actin dynamics is analyzed in vivo through the actin-based movement of the intracellular bacteria Shigella flexneri in cells expressing villin and its severing variant. The severing mutations abolish the gain of velocity induced by villin. To further analyze this effect, we reconstituted an in vitro actin-based bead movement in which the usual capping protein is replaced by either the wild type or the severing mutant of villin. Confirming the in vivo results, villin-severing activity enhances the velocity of beads by more than two-fold and reduces the density of actin in the comets. We propose a model in which, by severing actin filaments and capping their barbed ends, villin increases the concentration of actin monomers available for polymerization, a mechanism that might be paralleled in vivo when an enterocyte undergoes an epithelio-mesenchymal transition. PMID:17182858

  2. Ratiometric Imaging of the T-Cell Actin Cytoskeleton Reveals the Nature of Receptor-Induced Cytoskeletal Enrichment

    PubMed Central

    Smoligovets, Alexander A.; Smith, Adam W.; Groves, Jay T.

    2013-01-01

    The T-cell actin cytoskeleton mediates adaptive immune system responses to peptide antigens by physically directing the motion and clustering of T-cell receptors (TCRs) on the cell surface. When TCR movement is impeded by externally applied physical barriers, the actin network exhibits transient enrichment near the trapped receptors. The coordinated nature of the actin density fluctuations suggests that they are composed of filamentous actin, but it has not been possible to eliminate de novo polymerization at TCR-associated actin polymerizing factors as an alternative cause. Here, we use a dual-probe cytoskeleton labeling strategy to distinguish between stable and polymerizing pools of actin. Our results suggest that TCR-associated actin consists of a relatively high proportion of the stable cytoskeletal fraction and extends away from the cell membrane into the cell. This implies that actin enrichment at mechanically trapped TCRs results from three-dimensional bunching of the existing filamentous actin network. PMID:23931330

  3. Correlated time derivatives of current, electric field intensity, and magnetic flux density for triggered lightning at 15 m

    NASA Astrophysics Data System (ADS)

    Uman, M. A.; Schoene, J.; Rakov, V. A.; Rambo, K. J.; Schnetzer, G. H.

    2002-07-01

    We present measured current and its time derivative correlated with the corresponding electric field intensity and magnetic flux density and their time derivatives measured at 15 m for two lightning return strokes triggered in 1999 at Camp Blanding, Florida. Lightning was triggered to a vertical 2-m rod mounted at the center of a 70 × 70 m buried metallic grid. The rocket-launching system was located underground at the center of the grid. The experiment was designed to minimize any influence of either the strike object or a finite-conducting Earth (ground surface arcing and propagation effects) on the fields and field derivatives. The measured current derivative waveform and the return stroke portion of the magnetic flux density derivative and electric field intensity derivative waveforms associated with the two strokes are observed to be essentially unipolar pulses that have similar waveshapes for the first 150 ns or so, including the initial rising portion, the peak, and about 50 ns after the peak. The current and magnetic field derivative waveshapes are very similar for their total duration, and both decay to near zero about 200 ns after the peak derivative is reached. The electric field derivative decays more slowly than the current derivative after about 150 ns, taking about 500 ns to decay to near zero. The transmission-line model, the simplest available and most used return stroke model, is employed to calculate the return stroke field derivatives, given the measured current derivative as a model input, for return stroke speeds of 1 × 108 m s-1, 2 × 108 m s-1, and 3 × 108 m s-1 (the speed of light). A reasonable match between calculated and measured field derivative waveshapes is achieved for both strokes for a return stroke speed between 2 × 108 m s-1 and 3 × 108 m s-1. Although the measured field and current derivatives have similar waveshapes for about 150 ns, which might appear to be consistent with the hypothesis that the radiation field component

  4. Calcium-actin waves and oscillations of cellular membranes.

    PubMed

    Veksler, Alex; Gov, Nir S

    2009-09-16

    We propose a mechanism for the formation of membrane oscillations and traveling waves, which arise due to the coupling between the actin cytoskeleton and the calcium flux through the membrane. In our model, the fluid cell membrane has a mobile but constant population of proteins with a convex spontaneous curvature, which act as nucleators of actin polymerization and adhesion. Such a continuum model couples the forces of cell-substrate adhesion, actin polymerization, membrane curvature, and the flux of calcium through the membrane. Linear stability analysis shows that sufficiently strong coupling among the calcium, membrane, and protein dynamics may induce robust traveling waves on the membrane. This result was checked for a reduced feedback scheme and is compared to the results without the effects of calcium, where permanent phase separation without waves or oscillations is obtained. The model results are compared to the published observations of calcium waves in cell membranes, and a number of testable predictions are proposed. PMID:19751660

  5. Origin of dc voltage in type II superconducting flux pumps: field, field rate of change, and current density dependence of resistivity

    NASA Astrophysics Data System (ADS)

    Geng, J.; Matsuda, K.; Fu, L.; Fagnard, J.-F.; Zhang, H.; Zhang, X.; Shen, B.; Dong, Q.; Baghdadi, M.; Coombs, T. A.

    2016-03-01

    Superconducting flux pumps are the kind of devices which can generate direct current into superconducting circuit using external magnetic field. The key point is how to induce a dc voltage across the superconducting load by ac fields. Giaever (1966 IEEE Spectr. 3 117) pointed out flux motion in superconductors will induce a dc voltage, and demonstrated a rectifier model which depended on breaking superconductivity. van de Klundert et al (1981 Cryogenics 21 195, 267) in their review(s) described various configurations for flux pumps all of which relied on inducing the normal state in at least part of the superconductor. In this letter, following their work, we reveal that a variation in the resistivity of type II superconductors is sufficient to induce a dc voltage in flux pumps and it is not necessary to break superconductivity. This variation in resistivity is due to the fact that flux flow is influenced by current density, field intensity, and field rate of change. We propose a general circuit analogy for travelling wave flux pumps, and provide a mathematical analysis to explain the dc voltage. Several existing superconducting flux pumps which rely on the use of a travelling magnetic wave can be explained using the analysis enclosed. This work can also throw light on the design and optimization of flux pumps.

  6. A novel approach to calculate inductance and analyze magnetic flux density of helical toroidal coil applicable to Superconducting Magnetic Energy Storage systems (SMES)

    NASA Astrophysics Data System (ADS)

    Alizadeh Pahlavani, M. R.; Shoulaie, A.

    2010-04-01

    In this paper, formulas are proposed for the self and mutual inductance calculations of the helical toroidal coil (HTC) by the direct and indirect methods at superconductivity conditions. The direct method is based on the Neumann’s equation and the indirect approach is based on the toroidal and the poloidal components of the magnetic flux density. Numerical calculations show that the direct method is more accurate than the indirect approach at the expense of its longer computational time. Implementation of some engineering assumptions in the indirect method is shown to reduce the computational time without loss of accuracy. Comparison between the experimental measurements and simulated results for inductance, using the direct and the indirect methods indicates that the proposed formulas have high reliability. It is also shown that the self inductance and the mutual inductance could be calculated in the same way, provided that the radius of curvature is >0.4 of the minor radius, and that the definition of the geometric mean radius in the superconductivity conditions is used. Plotting contours for the magnetic flux density and the inductance show that the inductance formulas of helical toroidal coil could be used as the basis for coil optimal design. Optimization target functions such as maximization of the ratio of stored magnetic energy with respect to the volume of the toroid or the conductor’s mass, the elimination or the balance of stress in some coordinate directions, and the attenuation of leakage flux could be considered. The finite element (FE) approach is employed to present an algorithm to study the three-dimensional leakage flux distribution pattern of the coil and to draw the magnetic flux density lines of the HTC. The presented algorithm, due to its simplicity in analysis and ease of implementation of the non-symmetrical and three-dimensional objects, is advantageous to the commercial software such as ANSYS, MAXWELL, and FLUX. Finally, using the

  7. Hourly estimation of soil heat flux density at the soil surface with three models and two field methods

    NASA Astrophysics Data System (ADS)

    Venegas, P.; Grandón, A.; Jara, J.; Paredes, J.

    2013-04-01

    Heat flux density at the soil surface ( G 0) was evaluated hourly on a vegetal cover 0.08 m high, with a leaf area index of 1.07 m2 m-2, during daylight hours, using Choudhury et al. (Agric For Meteorol 39:283-297, 1987) ( G_0^{{rn}} ), Santanello and Friedl (J Appl Meteorol 42:851-862, 2003) ( G_0^{{s}} ), and force-restore ( G_0^{{fr}} ) models and the plate calorimetry methodology ( G_0^{{pco}} ), where the gradient calorimetry methodology ( G 0 R ) served as a reference for determining G 0. It was found that the peak of G 0R was at 1 p.m., with values that ranged between 60 and 100 W m-2 and that the G 0/Rn relation varied during the day with values close to zero in the early hours of the morning and close to 0.25 in the last hours of daylight. The G_0^{{s}} model presented the best performance, followed by the G_0^{{rn}} and G_0^{{fr}} models. The plate calorimetry methodology showed a similar behavior to that of the gradient calorimetry referential methodology.

  8. Inverse relationship between photon flux densities and nanotesla magnetic fields over cell aggregates: Quantitative evidence for energetic conservation

    PubMed Central

    Persinger, Michael A.; Dotta, Blake T.; Karbowski, Lukasz M.; Murugan, Nirosha J.

    2015-01-01

    The quantitative relationship between local changes in magnetic fields and photon emissions within ∼2 mm of aggregates of 105–106 cells was explored experimentally. The vertical component of the earth’s magnetic field as measured by different magnetometers was ∼15 nT higher when plates of cells removed from incubation were measured compared to plates containing only medium. Additional experiments indicated an inverse relationship over the first ∼45 min between changes in photon counts (∼10−12 W·m−2) following removal from incubation and similar changes in magnetic field intensity. Calculations indicated that the energy within the aqueous volume containing the cells was equivalent for that associated with the flux densities of the magnetic fields and the photon emissions. For every approximately 1 nT increase in magnetic field intensity value there was a decrease of ∼2 photons (equivalent of 10−18 J). These results complement correlation studies and suggest there may be a conservation of energy between expression as magnetic fields that are subtracted or added to the adjacent geomagnetic field and reciprocal changes in photon emissions when aggregates of cells within a specific volume of medium (water) adapt to new environments. PMID:26005634

  9. Superfluid Density and Flux-Flow Resistivity Measurements of Multiple-Band Superconductor β-PdBi2

    NASA Astrophysics Data System (ADS)

    Okada, Tatsunori; Imai, Yoshinori; Maeda, Atsutaka

    β -PdBi2 (Tcmax = 5 . 4 K) is a newcomer of the multiple-band superconductors, revealed by the specific heat and the upper critical field measurements, and the angle-resolved photoemission spectroscopy. In addition, authors of ref. observed the spin-polarized band dispersion and proposed that β-PdBi2 is a candidate of topological superconductor. However, there is less information on superconducting properties so far. In order to clarify the superconducting gap function, we measured the temperature (T) and magnetic field (B) dependence of microwave complex conductivity of β-PdBi2 single crystals. We found that the superfluid density exhibits the thermally activated T dependence, manifesting the absence of nodes in the superconducting gaps. We also found that the flux-flow resistivity increased with B with downward-convex shape. Based on some theories, we considered that such a behavior originated from the backflow of supercurrents around vortices reflecting rather small Ginzburg-Landau parameter (κ ~= 5). This work was supported by the JSPS KAKENHI (Grant Numbers 15K17697 and 26-9315), and the JSPS Research Fellowship for Young Scientists.

  10. 5 to 160 keV continuous-wave x-ray spectral energy distribution and energy flux density measurements

    SciTech Connect

    Tallon, R.W.; Koller, D.C.; Pelzl, R.M.; Pugh, R.D.; Bellem, R.D. . Microelectronics and Photonics Research Branch)

    1994-12-01

    In 1991, the USAF Phillips Laboratory Microelectronics and Photonics Research Branch installed a low energy x-ray facility (LEXR) for use in microelectronics radiation-effects analysis and research. Techniques developed for measuring the x-ray spectral energy distribution (differential intensity) from a tungsten-target bremsstrahlung x-ray source are reported. Spectra with end-point energies ranging from 20 to 160 keV were recorded. A separate effort to calibrate the dosimetry for the Phillips Laboratory low-energy x-ray facility established a need to know the spectral energy distributions at some point within the facility (previous calibration efforts had relies on spectra obtained from computer simulations). It was discovered that the primary discrepancy between the simulated and measured spectra was in the L- K-line data. The associated intensity (energy flux density) of the measured distributions was found to be up to 30% higher. Based on the measured distributions, predicted device responses were within 10% of the measured response as compared to about 30% accuracy obtained with simulated distributions.

  11. EFFECTS OF ULTRAVIOLET-B IRRADIANCE ON SOYBEAN. V. THE DEPENDENCE OF PLANT SENSITIVITY ON THE PHOTOSYNTHETIC PHOTON FLUX DENSITY DURING AND AFTER LEAF EXPANSION

    EPA Science Inventory

    Soybeans (Glycine max (L.) Merr. cv Essex) were grown in a green house, and the first trifoliate leaf was either allowed to expand under a high photosynthetic photon flux density (PPFD) (1.4 millimoled per square meter per second) or a low PPFD (0.8 Millimoles per square meter pe...

  12. Plant lighting system with five wavelength-band light-emitting diodes providing photon flux density and mixing ratio control

    PubMed Central

    2012-01-01

    Background Plant growth and development depend on the availability of light. Lighting systems therefore play crucial roles in plant studies. Recent advancements of light-emitting diode (LED) technologies provide abundant opportunities to study various plant light responses. The LED merits include solidity, longevity, small element volume, radiant flux controllability, and monochromaticity. To apply these merits in plant light response studies, a lighting system must provide precisely controlled light spectra that are useful for inducing various plant responses. Results We have developed a plant lighting system that irradiated a 0.18 m2 area with a highly uniform distribution of photon flux density (PFD). The average photosynthetic PFD (PPFD) in the irradiated area was 438 micro-mol m–2 s–1 (coefficient of variation 9.6%), which is appropriate for growing leafy vegetables. The irradiated light includes violet, blue, orange-red, red, and far-red wavelength bands created by LEDs of five types. The PFD and mixing ratio of the five wavelength-band lights are controllable using a computer and drive circuits. The phototropic response of oat coleoptiles was investigated to evaluate plant sensitivity to the light control quality of the lighting system. Oat coleoptiles irradiated for 23 h with a uniformly distributed spectral PFD (SPFD) of 1 micro-mol m–2 s–1 nm–1 at every peak wavelength (405, 460, 630, 660, and 735 nm) grew almost straight upwards. When they were irradiated with an SPFD gradient of blue light (460 nm peak wavelength), the coleoptiles showed a phototropic curvature in the direction of the greater SPFD of blue light. The greater SPFD gradient induced the greater curvature of coleoptiles. The relation between the phototropic curvature (deg) and the blue-light SPFD gradient (micro-mol m–2 s–1 nm–1 m–1) was 2 deg per 1 micro-mol m–2 s–1 nm–1 m–1. Conclusions The plant lighting system, with a computer with a graphical user interface

  13. Assembly and Turnover of Short Actin Filaments by the Formin INF2 and Profilin*

    PubMed Central

    Gurel, Pinar S.; A, Mu; Guo, Bingqian; Shu, Rui; Mierke, Dale F.; Higgs, Henry N.

    2015-01-01

    INF2 (inverted formin 2) is a formin protein with unique biochemical effects on actin. In addition to the common formin ability to accelerate actin nucleation and elongation, INF2 can also sever filaments and accelerate their depolymerization. Although we understand key attributes of INF2-mediated severing, we do not understand the mechanism by which INF2 accelerates depolymerization subsequent to severing. Here, we show that INF2 can create short filaments (<60 nm) that continuously turn over actin subunits through a combination of barbed end elongation, severing, and WH2 motif-mediated depolymerization. This pseudo-steady state condition occurs whether starting from actin filaments or monomers. The rate-limiting step of the cycle is nucleotide exchange of ADP for ATP on actin monomers after release from the INF2/actin complex. Profilin addition has two effects: 1) to accelerate filament turnover 6-fold by accelerating nucleotide exchange and 2) to shift the equilibrium toward polymerization, resulting in longer filaments. In sum, our findings show that the combination of multiple interactions of INF2 with actin can work in concert to increase the ATP turnover rate of actin. Depending on the ratio of INF2:actin, this increased flux can result in rapid filament depolymerization or maintenance of short filaments. We also show that high concentrations of cytochalasin D accelerate ATP turnover by actin but through a different mechanism from that of INF2. PMID:26124273

  14. Photosynthetic response of Cannabis sativa L. to variations in photosynthetic photon flux densities, temperature and CO2 conditions.

    PubMed

    Chandra, Suman; Lata, Hemant; Khan, Ikhlas A; Elsohly, Mahmoud A

    2008-10-01

    Effect of different photosynthetic photon flux densities (0, 500, 1000, 1500 and 2000 μmol m(-2)s(-1)), temperatures (20, 25, 30, 35 and 40 °C) and CO2 concentrations (250, 350, 450, 550, 650 and 750 μmol mol(-1)) on gas and water vapour exchange characteristics of Cannabis sativa L. were studied to determine the suitable and efficient environmental conditions for its indoor mass cultivation for pharmaceutical uses. The rate of photosynthesis (PN) and water use efficiency (WUE) of Cannabis sativa increased with photosynthetic photon flux densities (PPFD) at the lower temperatures (20-25 °C). At 30 °C, PN and WUE increased only up to 1500 μmol m(-2)s(-1) PPFD and decreased at higher light levels. The maximum rate of photosynthesis (PN max) was observed at 30 °C and under 1500 μmol m(-2)s(-1) PPFD. The rate of transpiration (E) responded positively to increased PPFD and temperature up to the highest levels tested (2000 μmol m(-2)s(-1) and 40 °C). Similar to E, leaf stomatal conductance (gs) also increased with PPFD irrespective of temperature. However, gs increased with temperature up to 30 °C only. Temperature above 30 °C had an adverse effect on gs in this species. Overall, high temperature and high PPFD showed an adverse effect on PN and WUE. A continuous decrease in intercellular CO2 concentration (Ci) and therefore, in the ratio of intercellular CO2 to ambient CO2 concentration (Ci/Ca) was observed with the increase in temperature and PPFD. However, the decrease was less pronounced at light intensities above 1500 μmol m(-2)s(-1). In view of these results, temperature and light optima for photosynthesis was concluded to be at 25-30 °C and ∼1500 μmol m(-2)s(-1) respectively. Furthermore, plants were also exposed to different concentrations of CO2 (250, 350, 450, 550, 650 and 750 μmol mol(-1)) under optimum PPFD and temperature conditions to assess their photosynthetic response. Rate of photosynthesis, WUE and Ci decreased by 50 %, 53 % and 10

  15. Influence of soft ferromagnetic sections on the magnetic flux density profile of a large grain, bulk Y-Ba-Cu-O superconductor

    NASA Astrophysics Data System (ADS)

    Philippe, M. P.; Ainslie, M. D.; Wéra, L.; Fagnard, J.-F.; Dennis, A. R.; Shi, Y.-H.; Cardwell, D. A.; Vanderheyden, B.; Vanderbemden, P.

    2015-09-01

    Bulk, high temperature superconductors have significant potential for use as powerful permanent magnets in a variety of practical applications due to their ability to trap record magnetic fields. In this paper, soft ferromagnetic sections are combined with a bulk, large grain Y-Ba-Cu-O high temperature superconductor to form superconductor/ferromagnet hybrid structures. We study how the ferromagnetic sections influence the shape of the profile of the trapped magnetic induction at the surface of each structure and report the surface magnetic flux density measured by Hall probe mapping. These configurations have been modelled using a 2D axisymmetric finite element method based on the H -formulation and the results show excellent qualitative and quantitative agreement with the experimental measurements. The model has also been used to study the magnetic flux distribution and predict the behaviour for other constitutive laws and geometries. The results show that the ferromagnetic material acts as a magnetic shield, but the flux density and its gradient are enhanced on the face opposite to the ferromagnet. The thickness and saturation magnetization of the ferromagnetic material are important and a characteristic ferromagnet thickness d* is derived: below d*, saturation of the ferromagnet occurs, and above d*, a weak thickness-dependence is observed. The influence of the ferromagnet is observed even if its saturation magnetization is lower than the trapped flux density of the superconductor. Conversely, thin ferromagnetic discs can be driven to full saturation even though the outer magnetic field is much smaller than their saturation magnetization.

  16. Actin-based phagosome motility.

    PubMed

    Zhang, Fangliang; Southwick, Frederick S; Purich, Daniel L

    2002-10-01

    Despite abundant evidence of actin's involvement at the particle internalization stage of phagocytosis, little is known about whether phagosomes undergo the same type of actin-based motility as observed with endocytic vesicles or such intracellular pathogens as Listeria and Shigella. By employing video microscopy to follow the fate of latex bead-containing phagosomes within the cytoplasm of bone marrow macrophages, we have made the novel observation of actin-based phagosome motility. Immunofluorescence microscopy confirmed that phagosomes containing IgG-opsonized, bovine serum albumin (or BSA) -coated or uncoated latex beads all formed actin-rich rocket tails that persisted only during a brief, 1-2 min period of actin-based motility. Average speeds of actin-based phagosome motility were 0.13 +/- 0.06 microm/s for IgG-coated beads, 0.14 +/- 0.04 microm/s for BSA-coated beads, and 0.11+/- 0.03 microm/s for uncoated beads. Moreover, the speeds and motile-phase duration of each type of phagosome were comparable to the behavior of pinosomes [Merrifield et al., 1999: Nat. Cell Biol. 1:72-74.]. Determination of optimal conditions for observing and analyzing actin-based phagosome motility should facilitate future investigations of phagocytosis and phagosome maturation. PMID:12211106

  17. Formin' actin in the nucleus.

    PubMed

    Baarlink, Christian; Grosse, Robert

    2014-01-01

    Many if not most proteins can, under certain conditions, change cellular compartments, such as, for example, shuttling from the cytoplasm to the nucleus. Thus, many proteins may exert functions in various and very different subcellular locations, depending on the signaling context. A large amount of actin regulatory proteins has been detected in the mammalian cell nucleus, although their potential roles are much debated and are just beginning to emerge. Recently, members of the formin family of actin nucleators were also reported to dynamically localize to the nuclear environment. Here we discuss our findings that specific diaphanous-related formins can promote nuclear actin assembly in a signal-dependent manner. PMID:24637338

  18. Bacterial actins and their diversity

    PubMed Central

    Ozyamak, Ertan; Kollman, Justin M.; Komeili, Arash

    2015-01-01

    For many years bacteria were considered rather simple organisms, but the dogmatic notion that subcellular organization is a eukaryotic trait has been overthrown for more than a decade. The discovery of homologs of the eukaryotic cytoskeletal proteins actin, tubulin, and intermediate filaments in bacteria has been instrumental in changing this view. Over the recent years we gained an incredible level of insight into the diverse family of bacterial actins and their molecular workings. Here we review the functional, biochemical and structural features of the most well-studied bacterial actins. PMID:24015924

  19. Assessing variation in the radial profile of sap flux density in Pinus species and its effect on daily water use.

    PubMed

    Ford, Chelcy R; McGuire, Mary Anne; Mitchell, Robert J; Teskey, Robert O

    2004-03-01

    We monitored sap flux density (v) diurnally in nine mature southeastern pine (Pinus spp.) trees with a thermal dissipation probe that spanned the sapwood radius. We found the expected pattern of high v near the cambium and decreasing v with depth toward the center of the tree; however, the pattern was not constant within a day or between trees. Radial profiles of trees were steeper earlier in the day and became less steep later in the day. As a result, time-dependent changes in the shape of the radial profile of v were sometimes correlated with daily changes in evaporative demand. As the radial profile became less steep, the inner xylem contributed relatively more to total tree sap flow than it did earlier in the day. We present a 3-parameter Gaussian function that can be used to describe the radial distribution of v in trees. Parameters in the function represent depth in the xylem from the cambium, maximum v, depth in the xylem where maximum v occurs, and the rate of radial change in v with radial depth (beta). Values of beta varied significantly between trees and with time, and were sometimes correlated with air vapor pressure deficit (D). We hypothesize that this occurred during periods of high transpiration when the water potential gradient became great enough to move water in the inner sapwood despite its probable high hydraulic resistance. We examined discrepancies among estimates of daily water use based on single-point, two-point and multi-point (i.e., every 20 mm in the sapwood) measurements. When radial distribution of v was not considered, a single-point measurement resulted in errors as large as 154% in the estimate of daily water use relative to the estimate obtained from a multi-point measurement. Measuring v at two close sample points (10 and 30 mm) did not improve the estimate; however, estimates derived from v measured at two distant sample points (10 and 70 mm) significantly improved the estimate of daily water use, although errors were as great

  20. Comparison of the quantum and classical calculations of flux density of (220) channeled positrons in Si crystal

    NASA Astrophysics Data System (ADS)

    Korotchenko, K. B.; Tukhfatullin, TA; Pivovarov, Yu L.; Eikhorn, Yu L.

    2016-07-01

    Simulation of flux-peaking effect of the 255 MeV positrons channeled in (220) Si crystals is performed in the frame of classical and quantum mechanics. Comparison of the results obtained using both approaches shows relatively good agreement.

  1. Two new methods used to simulate the circumferential solar flux density concentrated on the absorber of a parabolic trough solar collector

    NASA Astrophysics Data System (ADS)

    Guo, Minghuan; Wang, Zhifeng; Sun, Feihu

    2016-05-01

    The optical efficiencies of a solar trough concentrator are important to the whole thermal performance of the solar collector, and the outer surface of the tube absorber is a key interface of energy flux. So it is necessary to simulate and analyze the concentrated solar flux density distributions on the tube absorber of a parabolic trough solar collector for various sun beam incident angles, with main optical errors considered. Since the solar trough concentrators are linear focusing, it is much of interest to investigate the solar flux density distribution on the cross-section profile of the tube absorber, rather than the flux density distribution along the focal line direction. Although a few integral approaches based on the "solar cone" concept were developed to compute the concentrated flux density for some simple trough concentrator geometries, all those integral approaches needed special integration routines, meanwhile, the optical parameters and geometrical properties of collectors also couldn't be changed conveniently. Flexible Monte Carlo ray trace (MCRT) methods are widely used to simulate the more accurate concentrated flux density distribution for compound parabolic solar trough concentrators, while generally they are quite time consuming. In this paper, we first mainly introduce a new backward ray tracing (BRT) method combined with the lumped effective solar cone, to simulate the cross-section flux density on the region of interest of the tube absorber. For BRT, bundles of rays are launched at absorber-surface points of interest, directly go through the glass cover of the absorber, strike on the uniformly sampled mirror segment centers in the close-related surface region of the parabolic reflector, and then direct to the effective solar cone around the incident sun beam direction after the virtual backward reflection. All the optical errors are convoluted into the effective solar cone. The brightness distribution of the effective solar cone is supposed

  2. Development of a vector-tensor system to measure the absolute magnetic flux density and its gradient in magnetically shielded rooms

    SciTech Connect

    Voigt, J.; Knappe-Grüneberg, S.; Gutkelch, D.; Neuber, S.; Schnabel, A.; Burghoff, M.; Haueisen, J.

    2015-05-15

    Several experiments in fundamental physics demand an environment of very low, homogeneous, and stable magnetic fields. For the magnetic characterization of such environments, we present a portable SQUID system that measures the absolute magnetic flux density vector and the gradient tensor. This vector-tensor system contains 13 integrated low-critical temperature (LTc) superconducting quantum interference devices (SQUIDs) inside a small cylindrical liquid helium Dewar with a height of 31 cm and 37 cm in diameter. The achievable resolution depends on the flux density of the field under investigation and its temporal drift. Inside a seven-layer mu-metal shield, an accuracy better than ±23 pT for the components of the static magnetic field vector and ±2 pT/cm for each of the nine components of the gradient tensor is reached by using the shifting method.

  3. Temperature and density characteristics of the Helicity Injected Torus-II spherical tokamak indicating closed flux sustainment using coaxial helicity injection

    SciTech Connect

    Hamp, W. T.; Jarboe, T. R.; Nelson, B. A.; O'Neill, R. G.; Raman, R.; Redd, A. J.; Stewart, B. T.; Mueller, D.

    2008-08-15

    The electron temperature and density profiles of plasmas in the Helicity Injected Torus [HIT-II: T. R. Jarboe et al., Phys. Plasmas 5, 1807 (1998)] experiment are measured by multipoint Thomson scattering (MPTS). The HIT-II device is a small low-aspect-ratio tokamak (major radius 0.3 m, minor radius 0.2 m, toroidal field of up to 0.5 T), capable of inductive ohmic (OH) current drive, Coaxial Helicity Injection (CHI) current drive, or combinations of both. The temperature and density characteristics have been characterized by a ruby laser MPTS diagnostic at up to six locations within the plasma for a single diagnostic time per discharge. Observed hollow temperature profiles of CHI discharges are inconsistent with open flux only predictions for CHI and indicate a closed flux region during CHI current drive.

  4. Force Feedback Controls Motor Activity and Mechanical Properties of Self-Assembling Branched Actin Networks.

    PubMed

    Bieling, Peter; Li, Tai-De; Weichsel, Julian; McGorty, Ryan; Jreij, Pamela; Huang, Bo; Fletcher, Daniel A; Mullins, R Dyche

    2016-01-14

    Branched actin networks--created by the Arp2/3 complex, capping protein, and a nucleation promoting factor--generate and transmit forces required for many cellular processes, but their response to force is poorly understood. To address this, we assembled branched actin networks in vitro from purified components and used simultaneous fluorescence and atomic force microscopy to quantify their molecular composition and material properties under various forces. Remarkably, mechanical loading of these self-assembling materials increases their density, power, and efficiency. Microscopically, increased density reflects increased filament number and altered geometry but no change in average length. Macroscopically, increased density enhances network stiffness and resistance to mechanical failure beyond those of isotropic actin networks. These effects endow branched actin networks with memory of their mechanical history that shapes their material properties and motor activity. This work reveals intrinsic force feedback mechanisms by which mechanical resistance makes self-assembling actin networks stiffer, stronger, and more powerful. PMID:26771487

  5. Ubiquitin ligase TRIM3 controls hippocampal plasticity and learning by regulating synaptic γ-actin levels

    PubMed Central

    Schreiber, Joerg; Végh, Marlene J.; Dawitz, Julia; Kroon, Tim; Loos, Maarten; Labonté, Dorthe; Li, Ka Wan; Van Nierop, Pim; Van Diepen, Michiel T.; De Zeeuw, Chris I.; Kneussel, Matthias; Meredith, Rhiannon M.; Smit, August B.

    2015-01-01

    Synaptic plasticity requires remodeling of the actin cytoskeleton. Although two actin isoforms, β- and γ-actin, are expressed in dendritic spines, the specific contribution of γ-actin in the expression of synaptic plasticity is unknown. We show that synaptic γ-actin levels are regulated by the E3 ubiquitin ligase TRIM3. TRIM3 protein and Actg1 transcript are colocalized in messenger ribonucleoprotein granules responsible for the dendritic targeting of messenger RNAs. TRIM3 polyubiquitylates γ-actin, most likely cotranslationally at synaptic sites. Trim3−/− mice consequently have increased levels of γ-actin at hippocampal synapses, resulting in higher spine densities, increased long-term potentiation, and enhanced short-term contextual fear memory consolidation. Interestingly, hippocampal deletion of Actg1 caused an increase in long-term fear memory. Collectively, our findings suggest that temporal control of γ-actin levels by TRIM3 is required to regulate the timing of hippocampal plasticity. We propose a model in which TRIM3 regulates synaptic γ-actin turnover and actin filament stability and thus forms a transient inhibitory constraint on the expression of hippocampal synaptic plasticity. PMID:26527743

  6. Tobacco Arp3 is localized to actin-nucleating sites in vivo

    PubMed Central

    Maisch, Jan; Fišerová, Jindřiška; Fischer, Lukáš; Nick, Peter

    2009-01-01

    The polarity of actin is a central determinant of intracellular transport in plant cells. To visualize actin polarity in living plant cells, the tobacco homologue of the actin-related protein 3 (ARP3) was cloned and a fusion with the red fluorescent protein (RFP) was generated. Upon transient expression of these fusions in the tobacco cell line BY-2 (Nicotiana tabacum L. cv. Bright Yellow 2), punctate structures were observed near the nuclear envelope and in the cortical plasma. These dots could be shown to decorate actin filaments by expressing RFP–ARP3 in a marker line, where actin was tagged by GFP (green fluorescent protein)–FABD (fimbrin actin-binding domain 2). When actin filaments were disrupted by latrunculin B or by prolonged cold treatment, and subsequently allowed to recover, the actin filaments reformed from the RFP–ARP3 structures, that therefore represented actin nucleation sites. The intracellular distribution of these sites was followed during the formation of pluricellular files, and it was observed that the density of RFP–ARP3 increased in the apex of the polarized, terminal cells of a file, whereas it was equally distributed in the central cells of a file. These findings are interpreted in terms of position-dependent differences of actin organization. PMID:19129161

  7. ACD toxin-produced actin oligomers poison formin-controlled actin polymerization

    PubMed Central

    Heisler, David B.; Kudryashova, Elena; Grinevich, Dmitry O.; Suarez, Cristian; Winkelman, Jonathan D.; Birukov, Konstantin G.; Kotha, Sainath R.; Parinandi, Narasimham L.; Vavylonis, Dimitrios; Kovar, David R.; Kudryashov, Dmitri S.

    2015-01-01

    The actin crosslinking domain (ACD) is an actin-specific toxin produced by several pathogens, including life-threatening spp. of Vibrio cholerae, Vibrio vulnificus, and Aeromonas hydrophila. Actin crosslinking by ACD is thought to lead to slow cytoskeleton failure owing to a gradual sequestration of actin in the form of nonfunctional oligomers. Here we found that ACD converted cytoplasmic actin into highly toxic oligomers that potently “poisoned” the ability of major actin assembly proteins, formins, to sustain actin polymerization. Thus, ACD can target the most abundant cellular protein by employing actin oligomers as secondary toxins to efficiently subvert cellular functions of actin while functioning at very low doses. PMID:26228148

  8. Final report on P1-APMP.EM-S9: VNIIM/KRISS bilateral comparison of DC magnetic flux density by means of a transfer standard coil

    NASA Astrophysics Data System (ADS)

    Shifrin, V. Ya; Park, P. G.

    2013-01-01

    The purpose of this bilateral comparison is to check the conformance of the base quantities of magnetic measurements, DC magnetic flux density and its ratio to a current, as reproduced at VNIIM and KRISS. In these institutes adequate conditions for precise measurements in low magnetic fields are provided and the appropriate equipment for attaining a high level of accuracy is available. The results in this report cover the comparisons of two units, T/A and T, reproduced by the two institutes. The experimental comparison data show good agreement within the estimated uncertainty components of the standards. The coordinated values of the unit of DC magnetic flux density and its ratios to DC current show a standard uncertainty at the level of 1 × 10-6 to 1.2 × 10-6 (k = 1) using the value of the gyromagnetic ratio of the shielded protons γp that was recommended by CODATA in 2010, the experimental determination of the ratio (γ4He/γp) of 4He atoms to protons, and the standards of the two institutes. The results give a basis for carrying out multilateral comparisons of standard quantum magnetometers of metrological institutes in the framework of APMP with participation of geomagnetic observatories, which require the establishment of a unified standard of the unit of DC magnetic flux density. They also show the possibility of decreasing the uncertainty of the determination of the unit of DC magnetic flux density from direct comparisons of standard quantum magnetometers. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by APMP, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  9. Epidemiology of actinic keratoses.

    PubMed

    Green, Adèle C

    2015-01-01

    The epidemiology of actinic keratoses (AKs) reflects their causation by cumulative sun exposure, with the highest prevalence seen in pale-skinned people living at low latitudes and on the most sun-exposed body sites, namely the hands, forearms and face. AKs are markers of increased risk of basal cell carcinoma, squamous cell carcinoma and melanoma, especially when they are numerous and have coalesced into an area of 'field cancerisation'. The major risk factors are male sex, advanced age, sun-sensitive complexion, high lifetime sun exposure and prolonged immunosuppression. Clinical counts of AKs enable the assessment and monitoring of AK burden, but accurate counting is notoriously difficult, especially when skin is severely sun damaged. AK counting has been repeatedly shown to be unreliable, even among expert dermatologists. Notwithstanding these challenges, qualitative assessment of the natural history of AKs shows a high turnover, with new lesions developing and with other lesions regressing. A very small proportion of AKs undergo malignant transformation, but the precise rate of transformation is unknown due to the inaccuracies in monitoring AK lesions over time. Primary prevention of AKs is achieved by limiting intense sun exposure through sun-protective behaviour, including seeking deep shade, wearing sun-protective clothing and applying sunscreen regularly to exposed skin, from an early age. PMID:25561199

  10. Chemotaxis and Actin Oscillations

    NASA Astrophysics Data System (ADS)

    Bodenschatz, Eberhard; Hsu, Hsin-Fang; Negrete, Jose; Beta, Carsten; Pumir, Alain; Gholami, Azam; Tarantola, Marco; Westendorf, Christian; Zykov, Vladimir

    Recently, self-oscillations of the cytoskeletal actin have been observed in Dictyostelium, a model system for studying chemotaxis. Here we report experimental results on the self-oscillation mechanism and the role of regulatory proteins and myosin II. We stimulate cells rapidly and periodically by using photo un-caging of the chemoattractant in a micro-fluidic device and measured the cellular responses. We found that the response amplitude grows with stimulation strength only in a very narrow region of stimulation, after which the response amplitude reaches a plateau. Moreover, the frequency-response is not constant but rather varies with the strength of external stimuli. To understand the underlying mechanism, we analyzed the polymerization and de-polymerization time in the single cell level. Despite of the large cell-to-cell variability, we found that the polymerization time is independent of external stimuli and the de-polymerization time is prolonged as the stimulation strength increases. Our conclusions will be summarized and the role of noise in the signaling network will be discussed. German Science Foundation CRC 937.

  11. Maximum limit to the number of myosin II motors participating in processive sliding of actin.

    PubMed

    Rastogi, Khushboo; Puliyakodan, Mohammed Shabeel; Pandey, Vikas; Nath, Sunil; Elangovan, Ravikrishnan

    2016-01-01

    In this work, we analysed processive sliding and breakage of actin filaments at various heavy meromyosin (HMM) densities and ATP concentrations in IVMA. We observed that with addition of ATP solution, the actin filaments fragmented stochastically; we then determined mean length and velocity of surviving actin filaments post breakage. Average filament length decreased with increase in HMM density at constant ATP, and increased with increase in ATP concentration at constant HMM density. Using density of HMM molecules and length of actin, we estimated the number of HMM molecules per actin filament (N) that participate in processive sliding of actin. N is solely a function of ATP concentration: 88 ± 24 and 54 ± 22 HMM molecules (mean ± S.D.) at 2 mM and 0.1 mM ATP respectively. Processive sliding of actin filament was observed only when N lay within a minimum lower limit (Nmin) and a maximum upper limit (Nmax) to the number of HMM molecules. When N < Nmin the actin filament diffused away from the surface and processivity was lost and when N > Nmax the filament underwent breakage eventually and could not sustain processive sliding. We postulate this maximum upper limit arises due to increased number of strongly bound myosin heads. PMID:27554800

  12. Maximum limit to the number of myosin II motors participating in processive sliding of actin

    PubMed Central

    Rastogi, Khushboo; Puliyakodan, Mohammed Shabeel; Pandey, Vikas; Nath, Sunil; Elangovan, Ravikrishnan

    2016-01-01

    In this work, we analysed processive sliding and breakage of actin filaments at various heavy meromyosin (HMM) densities and ATP concentrations in IVMA. We observed that with addition of ATP solution, the actin filaments fragmented stochastically; we then determined mean length and velocity of surviving actin filaments post breakage. Average filament length decreased with increase in HMM density at constant ATP, and increased with increase in ATP concentration at constant HMM density. Using density of HMM molecules and length of actin, we estimated the number of HMM molecules per actin filament (N) that participate in processive sliding of actin. N is solely a function of ATP concentration: 88 ± 24 and 54 ± 22 HMM molecules (mean ± S.D.) at 2 mM and 0.1 mM ATP respectively. Processive sliding of actin filament was observed only when N lay within a minimum lower limit (Nmin) and a maximum upper limit (Nmax) to the number of HMM molecules. When N < Nmin the actin filament diffused away from the surface and processivity was lost and when N > Nmax the filament underwent breakage eventually and could not sustain processive sliding. We postulate this maximum upper limit arises due to increased number of strongly bound myosin heads. PMID:27554800

  13. Simulation study of a geometric shape factor technique for estimating earth-emitted radiant flux densities from wide-field-of-view radiation measurements

    NASA Technical Reports Server (NTRS)

    Weaver, W. L.; Green, R. N.

    1980-01-01

    Geometric shape factors were computed and applied to satellite simulated irradiance measurements to estimate Earth emitted flux densities for global and zonal scales and for areas smaller than the detector field of view (FOV). Wide field of view flat plate detectors were emphasized, but spherical detectors were also studied. The radiation field was modeled after data from the Nimbus 2 and 3 satellites. At a satellite altitude of 600 km, zonal estimates were in error 1.0 to 1.2 percent and global estimates were in error less than 0.2 percent. Estimates with unrestricted field of view (UFOV) detectors were about the same for Lambertian and limb darkening radiation models. The opposite was found for restricted field of view detectors. The UFOV detectors are found to be poor estimators of flux density from the total FOV and are shown to be much better as estimators of flux density from a circle centered at the FOV with an area significantly smaller than that for the total FOV.

  14. Effect of texture and grain size on the magnetic flux density and core loss of cold-rolled high silicon steel sheets

    NASA Astrophysics Data System (ADS)

    Qin, Jing; Yang, Ping; Mao, Weimin; Ye, Feng

    2015-11-01

    The effects of texture and grain size on the magnetic flux density and core loss (50-20 kHz) of 0.23 mm-thick cold-rolled high silicon steel sheets are investigated by means of electron back-scattered diffraction (EBSD), loss separation, and anisotropy parameter (ε) calculation. A model of the hysteresis loss coefficient kh considering average grain size and ε is established. The magnetic flux density at 800 A/m (B8) is closely related to the volume fraction of η-fiber-oriented grains, while the magnetic flux density at 5000 A/m (B50) is closely related to the volume fractions of γ- and λ-fiber-oriented grains in high silicon steel. The hysteresis loss of high silicon steel can be greatly reduced by increasing the grain size and optimizing the texture of the sheets. Although increases in frequencies decrease the effect of texture on core loss, the effect cannot be ignored. As annealing temperature and time increase, the relative difference in core loss between the rolling direction (RD) and the transverse direction (TD) is maintained at higher frequencies because of increases in grain size, decreases in γ texture, and maintenance of a strong η texture. Texture and grain size jointly affect the high-frequency core loss of high silicon steel.

  15. Plant pathogenic bacteria target the actin microfilament network involved in the trafficking of disease defense components

    PubMed Central

    Jelenska, Joanna; Kang, Yongsung; Greenberg, Jean T

    2014-01-01

    Cells of infected organisms transport disease defense-related molecules along actin filaments to deliver them to their sites of action to combat the pathogen. To accommodate higher demand for intracellular traffic, plant F-actin density increases transiently during infection or treatment of Arabidopsis with pathogen-associated molecules. Many animal and plant pathogens interfere with actin polymerization and depolymerization to avoid immune responses. Pseudomonas syringae, a plant extracellular pathogen, injects HopW1 effector into host cells to disrupt the actin cytoskeleton and reduce vesicle movement in order to elude defense responses. In some Arabidopsis accessions, however, HopW1 is recognized and causes resistance via an actin-independent mechanism. HopW1 targets isoform 7 of vegetative actin (ACT7) that is regulated by phytohormones and environmental factors. We hypothesize that dynamic changes of ACT7 filaments are involved in plant immunity. PMID:25551177

  16. Identification of Arabidopsis Cyclase-associated Protein 1 as the First Nucleotide Exchange Factor for Plant Actin

    PubMed Central

    Chaudhry, Faisal; Guérin, Christophe; von Witsch, Matthias

    2007-01-01

    The actin cytoskeleton powers organelle movements, orchestrates responses to abiotic stresses, and generates an amazing array of cell shapes. Underpinning these diverse functions of the actin cytoskeleton are several dozen accessory proteins that coordinate actin filament dynamics and construct higher-order assemblies. Many actin-binding proteins from the plant kingdom have been characterized and their function is often surprisingly distinct from mammalian and fungal counterparts. The adenylyl cyclase-associated protein (CAP) has recently been shown to be an important regulator of actin dynamics in vivo and in vitro. The disruption of actin organization in cap mutant plants indicates defects in actin dynamics or the regulated assembly and disassembly of actin subunits into filaments. Current models for actin dynamics maintain that actin-depolymerizing factor (ADF)/cofilin removes ADP–actin subunits from filament ends and that profilin recharges these monomers with ATP by enhancing nucleotide exchange and delivery of subunits onto filament barbed ends. Plant profilins, however, lack the essential ability to stimulate nucleotide exchange on actin, suggesting that there might be a missing link yet to be discovered from plants. Here, we show that Arabidopsis thaliana CAP1 (AtCAP1) is an abundant cytoplasmic protein; it is present at a 1:3 M ratio with total actin in suspension cells. AtCAP1 has equivalent affinities for ADP– and ATP–monomeric actin (Kd ∼ 1.3 μM). Binding of AtCAP1 to ATP–actin monomers inhibits polymerization, consistent with AtCAP1 being an actin sequestering protein. However, we demonstrate that AtCAP1 is the first plant protein to increase the rate of nucleotide exchange on actin. Even in the presence of ADF/cofilin, AtCAP1 can recharge actin monomers and presumably provide a polymerizable pool of subunits to profilin for addition onto filament ends. In turnover assays, plant profilin, ADF, and CAP act cooperatively to promote flux of

  17. Regulation of water flow by actin-binding protein-induced actin gelatin.

    PubMed Central

    Ito, T; Suzuki, A; Stossel, T P

    1992-01-01

    Actin filaments inhibit osmotically driven water flow (Ito, T., K.S. Zaner, and T.P. Stossel. 1987. Biophys. J. 51: 745-753). Here we show that the actin gelation protein, actin-binding protein (ABP), impedes both osmotic shrinkage and swelling of an actin filament solution and reduces markedly the concentration of actin filaments required for this inhibition. These effects depend on actin filament immobilization, because the ABP concentration that causes initial impairment of water flow by actin filaments corresponds to the gel point measured viscometrically and because gelsolin, which noncovalently severs actin filaments, solates actin gels and restores water flow in a solution of actin cross-linked by ABP. Since ABP gels actin filaments in the periphery of many eukaryotic cells, such actin networks may contribute to physiological cell volume regulation. PMID:1318095

  18. Surface Renewal: An Advanced Micrometeorological Method for Measuring and Processing Field-Scale Energy Flux Density Data

    PubMed Central

    McElrone, Andrew J.; Shapland, Thomas M.; Calderon, Arturo; Fitzmaurice, Li; Paw U, Kyaw Tha; Snyder, Richard L.

    2013-01-01

    Advanced micrometeorological methods have become increasingly important in soil, crop, and environmental sciences. For many scientists without formal training in atmospheric science, these techniques are relatively inaccessible. Surface renewal and other flux measurement methods require an understanding of boundary layer meteorology and extensive training in instrumentation and multiple data management programs. To improve accessibility of these techniques, we describe the underlying theory of surface renewal measurements, demonstrate how to set up a field station for surface renewal with eddy covariance calibration, and utilize our open-source turnkey data logger program to perform flux data acquisition and processing. The new turnkey program returns to the user a simple data table with the corrected fluxes and quality control parameters, and eliminates the need for researchers to shuttle between multiple processing programs to obtain the final flux data. An example of data generated from these measurements demonstrates how crop water use is measured with this technique. The output information is useful to growers for making irrigation decisions in a variety of agricultural ecosystems. These stations are currently deployed in numerous field experiments by researchers in our group and the California Department of Water Resources in the following crops: rice, wine and raisin grape vineyards, alfalfa, almond, walnut, peach, lemon, avocado, and corn. PMID:24378712

  19. Surface renewal: an advanced micrometeorological method for measuring and processing field-scale energy flux density data.

    PubMed

    McElrone, Andrew J; Shapland, Thomas M; Calderon, Arturo; Fitzmaurice, Li; Paw U, Kyaw Tha; Snyder, Richard L

    2013-01-01

    Advanced micrometeorological methods have become increasingly important in soil, crop, and environmental sciences. For many scientists without formal training in atmospheric science, these techniques are relatively inaccessible. Surface renewal and other flux measurement methods require an understanding of boundary layer meteorology and extensive training in instrumentation and multiple data management programs. To improve accessibility of these techniques, we describe the underlying theory of surface renewal measurements, demonstrate how to set up a field station for surface renewal with eddy covariance calibration, and utilize our open-source turnkey data logger program to perform flux data acquisition and processing. The new turnkey program returns to the user a simple data table with the corrected fluxes and quality control parameters, and eliminates the need for researchers to shuttle between multiple processing programs to obtain the final flux data. An example of data generated from these measurements demonstrates how crop water use is measured with this technique. The output information is useful to growers for making irrigation decisions in a variety of agricultural ecosystems. These stations are currently deployed in numerous field experiments by researchers in our group and the California Department of Water Resources in the following crops: rice, wine and raisin grape vineyards, alfalfa, almond, walnut, peach, lemon, avocado, and corn. PMID:24378712

  20. Possible roles of actin and myosin during anaphase chromosome movements in locust spermatocytes.

    PubMed

    Fabian, Lacramioara; Forer, Arthur

    2007-01-01

    We tested whether the mechanisms of chromosome movement during anaphase in locust (Locusta migratoria L.) spermatocytes might be similar to those described for crane-fly spermatocytes. Actin and myosin have been implicated in anaphase chromosome movements in crane-fly spermatocytes, as indicated by the effects of inhibitors and by the localisations of actin and myosin in spindles. In this study, we tested whether locust spermatocyte spindles also utilise actin and myosin, and whether actin is involved in microtubule flux. Living locust spermatocytes were treated with inhibitors of actin (latrunculin B and cytochalasin D), myosin (BDM), or myosin phosphorylation (Y-27632 and ML-7). We added drugs (individually) during anaphase. Actin inhibitors alter anaphase: chromosomes either completely stop moving, slow, or sometimes accelerate. The myosin inhibitor, BDM, also alters anaphase: in most cases, the chromosomes drastically slow or stop. ML-7, an inhibitor of MLCK, causes chromosomes to stop, slow, or sometimes accelerate, similar to actin inhibitors. Y-27632, an inhibitor of Rho-kinase, drastically slows or stops anaphase chromosome movements. The effects of the drugs on anaphase movement are reversible: most of the half-bivalents resumed movement at normal speed after these drugs were washed out. Actin and myosin were present in the spindles in locations consistent with their possible involvement in force production. Microtubule flux along kinetochore fibres is an actin-dependent process, since LatB completely removes or drastically reduces the gap in microtubule acetylation at the kinetochore. These results suggest that actin and myosin are involved in anaphase chromosome movements in locust spermatocytes. PMID:17922265

  1. Boolean gates on actin filaments

    NASA Astrophysics Data System (ADS)

    Siccardi, Stefano; Tuszynski, Jack A.; Adamatzky, Andrew

    2016-01-01

    Actin is a globular protein which forms long polar filaments in the eukaryotic cytoskeleton. Actin networks play a key role in cell mechanics and cell motility. They have also been implicated in information transmission and processing, memory and learning in neuronal cells. The actin filaments have been shown to support propagation of voltage pulses. Here we apply a coupled nonlinear transmission line model of actin filaments to study interactions between voltage pulses. To represent digital information we assign a logical TRUTH value to the presence of a voltage pulse in a given location of the actin filament, and FALSE to the pulse's absence, so that information flows along the filament with pulse transmission. When two pulses, representing Boolean values of input variables, interact, then they can facilitate or inhibit further propagation of each other. We explore this phenomenon to construct Boolean logical gates and a one-bit half-adder with interacting voltage pulses. We discuss implications of these findings on cellular process and technological applications.

  2. A dichotomy in cortical actin and chemotactic actin activity between human memory and naive T cells contributes to their differential susceptibility to HIV-1 infection.

    PubMed

    Wang, Weifeng; Guo, Jia; Yu, Dongyang; Vorster, Paul J; Chen, WanJun; Wu, Yuntao

    2012-10-12

    Human memory and naive CD4 T cells can mainly be identified by the reciprocal expression of the CD45RO or CD45RA isoforms. In HIV-1 infection, blood CD45RO memory CD4 T cells are preferentially infected and serve as a major viral reservoir. The molecular mechanism dictating this differential susceptibility to HIV-1 remains largely obscure. Here, we report that the different susceptibility of memory and naive T cells to HIV is not determined by restriction factors such as Apobec3G or BST2. However, we observed a phenotypic distinction between human CD45RO and CD45RA resting CD4 T cells in their cortical actin density and actin dynamics. CD45RO CD4 T cells possess a higher cortical actin density and can be distinguished as CD45RO(+)Actin(high). In contrast, CD45RA T cells are phenotypically CD45RA(+)Actin(low). In addition, the cortical actin in CD45RO memory CD4 T cells is more dynamic and can respond to low dosages of chemotactic induction by SDF-1, whereas that of naive cells cannot, despite a similar level of the chemokine receptor CXCR4 present on both cells. We further demonstrate that this difference in the cortical actin contributes to their differential susceptibility to HIV-1; resting memory but not naive T cells are highly responsive to HIV-mediated actin dynamics that promote higher levels of viral entry and early DNA synthesis in resting memory CD4 T cells. Furthermore, transient induction of actin dynamics in resting naive T cells rescues HIV latent infection following CD3/CD28 stimulation. These results suggest a key role of chemotactic actin activity in facilitating HIV-1 latent infection of these T cell subsets. PMID:22879601

  3. Technical advance: identification of plant actin-binding proteins by F-actin affinity chromatography

    NASA Technical Reports Server (NTRS)

    Hu, S.; Brady, S. R.; Kovar, D. R.; Staiger, C. J.; Clark, G. B.; Roux, S. J.; Muday, G. K.

    2000-01-01

    Proteins that interact with the actin cytoskeleton often modulate the dynamics or organization of the cytoskeleton or use the cytoskeleton to control their localization. In plants, very few actin-binding proteins have been identified and most are thought to modulate cytoskeleton function. To identify actin-binding proteins that are unique to plants, the development of new biochemical procedures will be critical. Affinity columns using actin monomers (globular actin, G-actin) or actin filaments (filamentous actin, F-actin) have been used to identify actin-binding proteins from a wide variety of organisms. Monomeric actin from zucchini (Cucurbita pepo L.) hypocotyl tissue was purified to electrophoretic homogeneity and shown to be native and competent for polymerization to actin filaments. G-actin, F-actin and bovine serum albumin affinity columns were prepared and used to separate samples enriched in either soluble or membrane-associated actin-binding proteins. Extracts of soluble actin-binding proteins yield distinct patterns when eluted from the G-actin and F-actin columns, respectively, leading to the identification of a putative F-actin-binding protein of approximately 40 kDa. When plasma membrane-associated proteins were applied to these columns, two abundant polypeptides eluted selectively from the F-actin column and cross-reacted with antiserum against pea annexins. Additionally, a protein that binds auxin transport inhibitors, the naphthylphthalamic acid binding protein, which has been previously suggested to associate with the actin cytoskeleton, was eluted in a single peak from the F-actin column. These experiments provide a new approach that may help to identify novel actin-binding proteins from plants.

  4. Bacterial Actins? An Evolutionary Perspective

    NASA Technical Reports Server (NTRS)

    Doolittle, Russell F.; York, Amanda L.

    2003-01-01

    According to the conventional wisdom, the existence of a cytoskeleton in eukaryotes and its absence in prokaryotes constitute a fundamental divide between the two domains of life. An integral part of the dogma is that a cytoskeleton enabled an early eukaryote to feed upon prokaryotes, a consequence of which was the occasional endosymbiosis and the eventual evolution of organelles. Two recent papers present compelling evidence that actin, one of the principal components of a cytoskeleton, has a homolog in Bacteria that behaves in many ways like eukaryotic actin. Sequence comparisons reveml that eukaryotic actin and the bacterial homolog (mreB protein), unlike many other proteins common to eukaryotes and Bacteria, have very different and more highly extended evolutionary histories.

  5. Actin engine in immunological synapse.

    PubMed

    Piragyte, Indre; Jun, Chang-Duk

    2012-06-01

    T cell activation and function require physical contact with antigen presenting cells at a specialized junctional structure known as the immunological synapse. Once formed, the immunological synapse leads to sustained T cell receptor-mediated signalling and stabilized adhesion. High resolution microscopy indeed had a great impact in understanding the function and dynamic structure of immunological synapse. Trends of recent research are now moving towards understanding the mechanical part of immune system, expanding our knowledge in mechanosensitivity, force generation, and biophysics of cell-cell interaction. Actin cytoskeleton plays inevitable role in adaptive immune system, allowing it to bear dynamic and precise characteristics at the same time. The regulation of mechanical engine seems very complicated and overlapping, but it enables cells to be very sensitive to external signals such as surface rigidity. In this review, we focus on actin regulators and how immune cells regulate dynamic actin rearrangement process to drive the formation of immunological synapse. PMID:22916042

  6. The Actin Cytoskeleton as a Therapeutic Target for the Prevention of Relapse to Methamphetamine Use.

    PubMed

    Young, Erica J; Briggs, Sherri B; Miller, Courtney A

    2015-01-01

    A high rate of relapse is a defining characteristic of substance use disorder for which few treatments are available. Exposure to environmental cues associated with previous drug use can elicit relapse by causing the involuntary retrieval of deeply engrained associative memories that trigger a strong motivation to seek out drugs. Our lab is focused on identifying and disrupting mechanisms that support these powerful consolidated memories, with the goal of developing therapeutics. A particularly promising mechanism is regulation of synaptic dynamics by actin polymerization within dendritic spines. Emerging evidence indicates that memory is supported by structural and functional plasticity dendritic spines, for which actin polymerization is critical, and that prior drug use increases both spine and actin dynamics. Indeed we have found that inhibiting amygdala (AMY) actin polymerization immediately or twenty-four hours prior to testing disrupted methamphetamine (METH)-associated memories, but not food reward or fear memories. Furthermore, METH training increased AMY spine density which was reversed by actin depolymerization treatment. Actin dynamics were also shifted to a more dynamic state by METH training. While promising, actin polymerization inhibitors are not a viable therapeutic, as a multitude of peripheral process (e.g. cardiac function) rely on dynamic actin. For this reason, we have shifted our focus upstream of actin polymerization to nonmuscle myosin II. We and others have demonstrated that myosin IIb imparts a mechanical force that triggers spine actin polymerization in response to synaptic stimulation. Similar to an actin depolymerizing compound, pre-test inhibition of myosin II ATPase activity in the AMY produced a rapid and lasting disruption of drug-seeking behavior. While many questions remain, these findings indicate that myosin II represents a potential therapeutic avenue to target the actin cytoskeleton and disrupt the powerful, extinction

  7. Actin polymerization is stimulated by actin cross-linking protein palladin.

    PubMed

    Gurung, Ritu; Yadav, Rahul; Brungardt, Joseph G; Orlova, Albina; Egelman, Edward H; Beck, Moriah R

    2016-02-15

    The actin scaffold protein palladin regulates both normal cell migration and invasive cell motility, processes that require the co-ordinated regulation of actin dynamics. However, the potential effect of palladin on actin dynamics has remained elusive. In the present study, we show that the actin-binding immunoglobulin-like domain of palladin, which is directly responsible for both actin binding and bundling, also stimulates actin polymerization in vitro. Palladin eliminated the lag phase that is characteristic of the slow nucleation step of actin polymerization. Furthermore, palladin dramatically reduced depolymerization, slightly enhanced the elongation rate, and did not alter the critical concentration. Microscopy and in vitro cross-linking assays reveal differences in actin bundle architecture when palladin is incubated with actin before or after polymerization. These results suggest a model whereby palladin stimulates a polymerization-competent form of globular or monomeric actin (G-actin), akin to metal ions, either through charge neutralization or through conformational changes. PMID:26607837

  8. Hippocampal Dendritic Spines Are Segregated Depending on Their Actin Polymerization.

    PubMed

    Domínguez-Iturza, Nuria; Calvo, María; Benoist, Marion; Esteban, José Antonio; Morales, Miguel

    2016-01-01

    Dendritic spines are mushroom-shaped protrusions of the postsynaptic membrane. Spines receive the majority of glutamatergic synaptic inputs. Their morphology, dynamics, and density have been related to synaptic plasticity and learning. The main determinant of spine shape is filamentous actin. Using FRAP, we have reexamined the actin dynamics of individual spines from pyramidal hippocampal neurons, both in cultures and in hippocampal organotypic slices. Our results indicate that, in cultures, the actin mobile fraction is independently regulated at the individual spine level, and mobile fraction values do not correlate with either age or distance from the soma. The most significant factor regulating actin mobile fraction was the presence of astrocytes in the culture substrate. Spines from neurons growing in the virtual absence of astrocytes have a more stable actin cytoskeleton, while spines from neurons growing in close contact with astrocytes show a more dynamic cytoskeleton. According to their recovery time, spines were distributed into two populations with slower and faster recovery times, while spines from slice cultures were grouped into one population. Finally, employing fast lineal acquisition protocols, we confirmed the existence of loci with high polymerization rates within the spine. PMID:26881098

  9. Hippocampal Dendritic Spines Are Segregated Depending on Their Actin Polymerization

    PubMed Central

    Domínguez-Iturza, Nuria; Calvo, María; Benoist, Marion; Esteban, José Antonio; Morales, Miguel

    2016-01-01

    Dendritic spines are mushroom-shaped protrusions of the postsynaptic membrane. Spines receive the majority of glutamatergic synaptic inputs. Their morphology, dynamics, and density have been related to synaptic plasticity and learning. The main determinant of spine shape is filamentous actin. Using FRAP, we have reexamined the actin dynamics of individual spines from pyramidal hippocampal neurons, both in cultures and in hippocampal organotypic slices. Our results indicate that, in cultures, the actin mobile fraction is independently regulated at the individual spine level, and mobile fraction values do not correlate with either age or distance from the soma. The most significant factor regulating actin mobile fraction was the presence of astrocytes in the culture substrate. Spines from neurons growing in the virtual absence of astrocytes have a more stable actin cytoskeleton, while spines from neurons growing in close contact with astrocytes show a more dynamic cytoskeleton. According to their recovery time, spines were distributed into two populations with slower and faster recovery times, while spines from slice cultures were grouped into one population. Finally, employing fast lineal acquisition protocols, we confirmed the existence of loci with high polymerization rates within the spine. PMID:26881098

  10. Femtosecond pump-probe studies of actinic-wavelength dependence in aqueous chlorine dioxide photochemistry

    NASA Astrophysics Data System (ADS)

    Bixby, Teresa J.; Bolinger, Joshua C.; Patterson, Joshua D.; Reid, Philip J.

    2009-04-01

    The actinic or photolysis-wavelength dependence of aqueous chlorine dioxide (OClO) photochemistry is investigated using femtosecond pump-probe spectroscopy. Following photoexcitation at 310, 335, and 410 nm the photoinduced evolution in optical density is measured from the UV to the near IR. Analysis of the optical-density evolution illustrates that the quantum yield for atomic chlorine production (ΦCl) increases with actinic energy, with ΦCl=0.16±0.02 for 410 nm excitation and increasing to 0.25±0.01 and 0.54±0.10 for 335 and 310 nm excitations, respectively. Consistent with previous studies, the production of Cl occurs through two channels, with one channel corresponding to prompt (<5 ps) Cl formation and the other corresponding to the thermal decomposition of ClOO formed by OClO photoisomerization. The partitioning between Cl production channels is dependent on actinic energy, with prompt Cl production enhanced with an increase in actinic energy. Limited evidence is found for enhanced ClO production with an increase in actinic energy. Stimulated emission and excited-state absorption features associated with OClO populating the optically prepared A22 surface decrease with an increase in actinic energy suggesting that the excited-state decay dynamics are also actinic energy dependent. The studies presented here provide detailed information on the actinic-wavelength dependence of OClO photochemistry in aqueous solution.

  11. Femtosecond pump-probe studies of actinic-wavelength dependence in aqueous chlorine dioxide photochemistry

    SciTech Connect

    Bixby, Teresa J.; Bolinger, Joshua C.; Patterson, Joshua D.; Reid, Philip J.

    2009-04-21

    The actinic or photolysis-wavelength dependence of aqueous chlorine dioxide (OClO) photochemistry is investigated using femtosecond pump-probe spectroscopy. Following photoexcitation at 310, 335, and 410 nm the photoinduced evolution in optical density is measured from the UV to the near IR. Analysis of the optical-density evolution illustrates that the quantum yield for atomic chlorine production ({Phi}{sub Cl}) increases with actinic energy, with {Phi}{sub Cl}=0.16{+-}0.02 for 410 nm excitation and increasing to 0.25{+-}0.01 and 0.54{+-}0.10 for 335 and 310 nm excitations, respectively. Consistent with previous studies, the production of Cl occurs through two channels, with one channel corresponding to prompt (<5 ps) Cl formation and the other corresponding to the thermal decomposition of ClOO formed by OClO photoisomerization. The partitioning between Cl production channels is dependent on actinic energy, with prompt Cl production enhanced with an increase in actinic energy. Limited evidence is found for enhanced ClO production with an increase in actinic energy. Stimulated emission and excited-state absorption features associated with OClO populating the optically prepared {sup 2}A{sub 2} surface decrease with an increase in actinic energy suggesting that the excited-state decay dynamics are also actinic energy dependent. The studies presented here provide detailed information on the actinic-wavelength dependence of OClO photochemistry in aqueous solution.

  12. Follow-up observations at 16 and 33GHz of extragalactic sources from WMAP 3-yr data: II - Flux density variability

    NASA Astrophysics Data System (ADS)

    Franzen, Thomas M. O.; Davies, Matthew L.; Davies, Rod D.; Davis, Richard J.; Feroz, Farhan; Génova-Santos, Ricardo; Grainge, Keith J. B.; Green, David A.; Hobson, Michael P.; Hurley-Walker, Natasha; Lasenby, Anthony N.; López-Caniego, Marcos; Olamaie, Malak; Padilla-Torres, Carmen P.; Pooley, Guy G.; Rebolo, Rafael; Rodríguez-Gonzálvez, Carmen; Saunders, Richard D. E.; Scaife, Anna M. M.; Scott, Paul F.; Shimwell, Timothy W.; Titterington, David J.; Waldram, Elizabeth M.; Watson, Robert A.; Zwart, Jonathan T. L.

    2009-12-01

    Using the Arcminute Microkelvin Imager (AMI) at 16GHz and the Very Small Array (VSA) at 33GHz to make follow-up observations of sources in the New Extragalactic WMAP Point Source catalogue, we have investigated the flux density variability in a complete sample of 97 sources over time-scales of a few months to ~1.5yr. We find that 53 per cent of the 93 sources, for which we have multiple observations, are variable, at the 99 per cent confidence level, above the flux density calibration uncertainties of ~4 per cent at 16GHz the fraction of sources having varied by more than 20 per cent is 15 per cent at 16GHz and 20 per cent at 33GHz. Not only is this common occurrence of variability at high frequency of interest for source physics, but also strategies for coping with source contamination in cosmic microwave background work must take this variability into account. There is no strong evidence of a correlation between variability and flux density for the sample as a whole. For those sources classified as variable, the mean fractional rms variation in flux density increases significantly with the length of time separating observation pairs. Using a maximum likelihood method, we calculate the correlation in the variability at the two frequencies in a subset of sources classified as variable from both the AMI and VSA data and find the Pearson product-moment correlation coefficient to be very high (0.955 +/- 0.034). We also find the degree of variability at 16GHz (0.202 +/- 0.028) to be very similar to that at 33GHz (0.224 +/- 0.039). Finally, we have investigated the relationship between variability and spectral index, α33.7513.9 (where S ~ ν-α), and find a significant difference in the spectral indices of the variable sources (-0.06 +/- 0.05) and non-variable sources (0.13 +/- 0.04). We kindly request that any reference to this paper cites `AMI Consortium: Franzen et al. 2009'. Issuing author - email: t.franzen@mrao.cam.ac.uk ‡ E-mail: m.davies@mrao.cam.ac.uk

  13. Influence of the vapor flux on temperature, density, and abundance distributions in a multicomponent, porous, icy body

    NASA Technical Reports Server (NTRS)

    Benkhoff, J.; Huebner, W. F.

    1995-01-01

    We calculated the vapor flux of the icy components in the surface layer of a porous, short-period, Jupiter-class comet, in order to investigate the relationship of the observed relative molecular abundances in the coma with those in the nucleus. The model assumes a body containing one major ice component (H20) and up to three minor components of higher volatility (e.g., CO, CO2, CH3OH). The body's porous structure is modeled as a bundle of tubes with a given tortuosity and initially a constant pore diameter. The mass and energy equations for the different volatiles are solved simultaneously under appropiate boundary conditions. Heat is conducted by the matrix and carried by the vapors. The one-dimensional model includes radially inward and outward flowing vapor within the body, complete depletion of less volatile ices in outer layers, and recondensation of vapor in deeper, coller layers. As a result, we obtain the temperature and abundance distribution in the nucleus and the gas flux into the interior and into the coma for each of the volatiles at various positions in the orbit. The ratio of the gas flux of minor volatiles to that of H2) in the coma varies by several orders of magnitude throughout the orbit. Thus, the relative abundances of species observed in the coma are in most cases not the same as those in the nucleus. Results also indicate that it will be impossible to determine the relative abundances of ices more volatile than water from samples taken a few meters below the surface during a comet rendezvous mission. We made calculations for a wide range of different parameters, such as porosity, pore radius, and thermal conductivity of the matrix. To introduce the model we present typical results for a dust-free comet.

  14. Fluid-kinetic simulations of the passage of Storm Enhanced Density (SED) plasma flux tubes through the dayside cleft auroral processes region

    NASA Astrophysics Data System (ADS)

    Zeng, W.; Horwitz, J. L.

    2007-12-01

    Foster et al. [2002] and others have reported on elevated ionospheric density regions being convected from the subauroral plasmaspheric region toward noon, in association with convection of plasmaspheric tails in the dayside magnetosphere. It has been suggested that these so-called Storm Enhanced Density (SED) regions could serve as ionospheric plasma source populations for cleft ion fountain outflows. To investigate this scenario, we have used our Dynamic Fluid Kinetic (DyFK) model to simulate the entry of a high-density "plasmasphere-like" flux tube entering the cleft region and subjected to an episode of wave-driven transverse ion heating. We find that the O+ ion density at higher altitudes increases and the density at lower altitudes decreases, following this heating episode, indicating increased numbers of O+ ions from the ionospheric source gain sufficient energy to reach higher altitudes after the effects of transverse wave heating. We also find that O+- H+ crossing point in topside ionosphere moves upward as the wave heating continues. Foster, J. C., P. J. Erickson, A. J. Coster, J. Goldstein, and F. J. Rich, Ionospheric signatures of plasmaspheric tails, Geophys. Res. Lett., 29(13), 1623, doi:10.1029/2002GL015067, 2002.

  15. Effect of vertically aligned carbon nanotube density on the water flux and salt rejection in desalination membranes.

    PubMed

    Trivedi, Samarth; Alameh, Kamal

    2016-01-01

    In this paper, vertically aligned carbon nanotube (VACNT) membranes of different densities are developed and their performances are investigated. VACNT arrays of densities 5 × 10(9), 10(10), 5 × 10(10) and 10(11) tubes cm(-2), are initially grown on 1 cm × 1 cm silicon substrates using chemical vapour deposition. A VACNT membrane is realised by attaching a 300 μm-thick 1 cm × 1 cm VACNT array on silicon to a 4″ glass substrate, applying polydimethylsiloxane (PDMS) through spin coating to fill the gaps between the VACNTs, and using a microtome to slice the VACNT-PDMS composite into 25-μm-thick membranes. Experimental results show that the permeability of the developed VACNT membranes increases with the density of the VACNTs, while the salt rejection is almost independent of the VACNT density. The best measured permeance is attained with a VACNT membrane having a CNT density of 10(11) tubes cm(-2) is 1203 LMH at 1 bar. PMID:27504256

  16. Landscape Soil Respiration Fluxes are Related to Leaf Area Index, Stand Height and Density, and Soil Nitrogen in Rocky Mountain Subalpine Forests

    NASA Astrophysics Data System (ADS)

    Berryman, E.; Bradford, J. B.; Hawbaker, T. J.; Birdsey, R.; Ryan, M. G.

    2015-12-01

    There is a recent multi-agency push for accurate assessments of terrestrial carbon stocks and fluxes in the United States. Assessing the state of the carbon cycle in the US requires estimates of stocks and fluxes at large spatial scales. Such assessments are difficult, especially for soil respiration, which dominates ecosystem respiration and is notoriously highly variable over space and time. Here, we report three consecutive years of measurement of soil respiration fluxes in three 1 km2 subalpine forest landscapes: Fraser Experimental Forest (Colorado), Glacier Lakes Ecosystems Experimental Site ("GLEES", Wyoming), and Niwot Ridge (Colorado). Plots were established following the protocol of the US Forest Service's Forest Inventory and Analysis (FIA) Program. Clusters of plots were distributed across the landscape in a 0.25 km grid pattern. From 2004 through 2006, measurements of soil respiration were made once monthly during the growing season and twice during snowpack coverage for each year. Annual cumulative soil respiration was 6.10 (+/- 0.21) Mg ha-1y-1 for Fraser, 6.55 (+/- 0.27) Mg ha-1y-1 for GLEES, and 6.97 (+/- 0.20) Mg ha-1y-1 for Niwot. Variability in annual cumulative soil respiration varied by less than 20% among the three subalpine forests, despite differences in terrain, climate, disturbance history and anthropogenic nitrogen deposition. We quantified the relationship between respiration fluxes and commonly-measured forest properties and found that soil respiration was nonlinearly related to leaf area index, peaking around 2.5 m2m-2 then slowly declining. Annual litterfall (FA) was subtracted from soil respiration (FR) to calculate total belowground carbon flux (TBCF), which declined with increasing tree height, density and soil nitrogen. This landscape analysis of soil respiration confirmed experimentally-derived principles governing carbon fluxes in forests: as trees age and get taller, and in high-fertility areas, carbon flux to roots declines

  17. Magnetic-flux pump

    NASA Technical Reports Server (NTRS)

    Hildebrandt, A. F.; Elleman, D. D.; Whitmore, F. C. (Inventor)

    1966-01-01

    A magnetic flux pump is described for increasing the intensity of a magnetic field by transferring flux from one location to the magnetic field. The device includes a pair of communicating cavities formed in a block of superconducting material, and a piston for displacing the trapped magnetic flux into the secondary cavity producing a field having an intense flux density.

  18. Critical current densities and flux creep rates in near optimally doped BaFe2-xRuxAs2 (x≈0.7) single crystals

    NASA Astrophysics Data System (ADS)

    Haberkorn, N.; Eom, Man Jin; You, Jung Sang; Kim, Jeehoon; Kim, Jun Sung

    2016-04-01

    We present an investigation of the critical current densities Jc and flux creep rates in a near optimally doped BaFe2-xRuxAs2 (x≈0.7) single crystal by (measuring magnetization). The superconducting critical temperature is 18 K. The in-field dependences of the critical current density Jc are due to a mixed pinning scenario produced mainly by large precipitates and a less significant contribution of random disorder. Furthermore, a Maley analysis in the regime dominated by strong pinning centers (μ0H=0.1 T) is well described through a glassy exponent μ=1.9 and a collective pinning energy (U0) smaller than 100 K.

  19. Association of actin with alpha crystallins

    NASA Technical Reports Server (NTRS)

    Gopalakrishnan, S.; Boyle, D.; Takemoto, L.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    The alpha crystallins are cytosolic proteins that co-localize and co-purify with actin-containing microfilaments. Affinity column chromatography employing both covalently-coupled actin or alpha crystallin was used to demonstrate specific and saturable binding of actin with alpha crystallin. This conclusion was confirmed by direct visualization of alpha aggregates bound to actin polymerized in vitro. The significance of this interaction in relation to the functional properties of these two polypeptides will be discussed.

  20. An actin cytoskeleton with evolutionarily conserved functions in the absence of canonical actin-binding proteins

    PubMed Central

    Paredez, Alexander R.; Assaf, Zoe June; Sept, David; Timofejeva, Ljudmilla; Dawson, Scott C.; Wang, Chung-Ju Rachel; Cande, W. Z.

    2011-01-01

    Giardia intestinalis, a human intestinal parasite and member of what is perhaps the earliest-diverging eukaryotic lineage, contains the most divergent eukaryotic actin identified to date and is the first eukaryote known to lack all canonical actin-binding proteins (ABPs). We sought to investigate the properties and functions of the actin cytoskeleton in Giardia to determine whether Giardia actin (giActin) has reduced or conserved roles in core cellular processes. In vitro polymerization of giActin produced filaments, indicating that this divergent actin is a true filament-forming actin. We generated an anti-giActin antibody to localize giActin throughout the cell cycle. GiActin localized to the cortex, nuclei, internal axonemes, and formed C-shaped filaments along the anterior of the cell and a flagella-bundling helix. These structures were regulated with the cell cycle and in encysting cells giActin was recruited to the Golgi-like cyst wall processing vesicles. Knockdown of giActin demonstrated that giActin functions in cell morphogenesis, membrane trafficking, and cytokinesis. Additionally, Giardia contains a single G protein, giRac, which affects the Giardia actin cytoskeleton independently of known target ABPs. These results imply that there exist ancestral and perhaps conserved roles for actin in core cellular processes that are independent of canonical ABPs. Of medical significance, the divergent giActin cytoskeleton is essential and commonly used actin-disrupting drugs do not depolymerize giActin structures. Therefore, the giActin cytoskeleton is a promising drug target for treating giardiasis, as we predict drugs that interfere with the Giardia actin cytoskeleton will not affect the mammalian host. PMID:21444821

  1. Steady-state nuclear actin levels are determined by export competent actin pool.

    PubMed

    Skarp, Kari-Pekka; Huet, Guillaume; Vartiainen, Maria K

    2013-10-01

    A number of studies in the last decade have irrevocably promoted actin into a fully fledged member of the nuclear compartment, where it, among other crucial tasks, facilitates transcription and chromatin remodeling. Changes in nuclear actin levels have been linked to different cellular processes: decreased nuclear actin to quiescence and increased nuclear actin to differentiation. Importin 9 and exportin 6 transport factors are responsible for the continuous nucleocytoplasmic shuttling of actin, but the mechanisms, which result in modulated actin levels, have not been characterized. We find that in cells growing under normal growth conditions, the levels of nuclear actin vary considerably from cell to cell. To understand the basis for this, we have extensively quantified several cellular parameters while at the same time recording the import and export rates of green fluorescent protein (GFP)-tagged actin. Surprisingly, our dataset shows that the ratio of nuclear to cytoplasmic fluorescence intensity, but not nuclear shape, size, cytoplasm size, or their ratio, correlates negatively with both import and export rate of actin. This suggests that high-nuclear actin content is maintained by both diminished import and export. The high nuclear actin containing cells still show high mobility of actin, but it is not export competent, suggesting increased binding of actin to nuclear complexes. Creation of such export incompetent actin pool would ensure enough actin is retained in the nucleus and make it available for the various nuclear functions described for actin. PMID:23749625

  2. Actin Foci Adhesion of D. discoideum

    NASA Astrophysics Data System (ADS)

    Flanders, Bret; Paneru, Govind

    2014-03-01

    Amoeboid migration is a fast (10 μm min-1) integrin-independent mode of migration that is important with D. discoideum, leukocytes, and breast cancer cells. It is poorly understood, but depends on the establishment of adhesive contacts to the substrate where the cell transmits traction forces. In pre-aggregative D. discoideum, a model system for learning about amoeboid migration, these adhesive contacts are discrete complexes that are known as actin-foci. They have an area of ~ 0.5 μm2 and a lifetime of ~ 20 s. This talk will present measurements of the adhesive character of actin foci that have been obtained using a submicron force transducer that was designed for this purpose. Results on the rupture stresses and lifetimes of individual acting foci under nano-newton level forces will be described in the context of a general theory for cellular adhesion. This theory depends on, essentially, three cellular properties: the membrane-medium surface tension, the number density of adhesion receptors in the membrane, and the receptor-substrate potential energy surface. Therefore, the use of the transducer to determine the surface tension will be presented, as well.

  3. Actin dynamics: from nanoscale to microscale.

    PubMed

    Carlsson, Anders E

    2010-01-01

    The dynamic nature of actin in cells manifests itself constantly. Polymerization near the cell edge is balanced by depolymerization in the interior, externally induced actin polymerization is followed by depolymerization, and spontaneous oscillations of actin at the cell periphery are frequently seen. I discuss how mathematical modeling relates quantitative measures of actin dynamics to the rates of underlying molecular level processes. The dynamic properties addressed include the rate of actin assembly at the leading edge of a moving cell, the disassembly rates of intracellular actin networks, the polymerization time course in externally stimulated cells, and spontaneous spatiotemporal patterns formed by actin. Although several aspects of actin assembly have been clarified by increasingly sophisticated models, our understanding of rapid actin disassembly is limited, and the origins of nonmonotonic features in externally stimulated actin polymerization remain unclear. Theory has generated several concrete, testable hypotheses for the origins of spontaneous actin waves and cell-edge oscillations. The development and use of more biomimetic systems applicable to the geometry of a cell will be key to obtaining a quantitative understanding of actin dynamics in cells. PMID:20462375

  4. Effects of temperature, frequency, flux density, and excitation waveform on the core loss and dynamic B-H loops of Supermalloy

    SciTech Connect

    Schwarze, G.E.; Wieserman, W.R.; Niedra, J.M.

    1995-12-31

    The availability of experimental data which characterize the performance of soft magnetic materials for the combined conditions of temperature and frequency over a wide flux density range for different types of excitation is almost nonexistent. An experimental investigation of an 80-20 Ni-Fe alloy (Supermalloy) was conducted over the temperature (T) range of 23 to 300 C, frequency (f) range of 1 to 50 kHz, and maximum flux densities (B{sub M}) from 0.1 T up to 0.7 T for both sine and square wave voltage excitation. The investigation focused on the effects of B{sub M}, f, T, and excitation waveform on the specific core loss (SCL) and dynamic B-H loops. The results show that the ratio (R) of sine to square wave excitation specific core loss was always greater than unity for a given f and T and identical values of B{sub M}. The values of R ranged from 1.07 to 1.34. The classical theory of core loss separation into a hysteresis and eddy current loss component was used to theoretically determine the lower and upper bounds on R, against which the experimental R-values were compared. The experimental R-values were also used to make a comparison of the core loss of a sine and square wave voltage driven transformer.

  5. Effects of temperature, frequency, flux density, and excitation waveform on the core loss and dynamic B-H loops of supermalloy

    NASA Technical Reports Server (NTRS)

    Schwarze, Gene E.; Wieserman, William R.; Niedra, Janis M.

    1995-01-01

    The availability of experimental data which characterize the performance of soft magnetic materials for the combined conditions of temperature and frequency over a wide flux density range for different types of excitation is almost nonexistent. An experimental investigation of an 80-20 Ni-Fe alloy (Supermalloy) was conducted over the temperature (T) range of 23 to 300 C, frequency (f) range of 1 to 50 kHz, and maximum flux densities (B(sub M)) from 0.1 T up to 0.7 T for both sine and square wave voltage excitation. The investigation focused on the effects of (B(sub M)), f, T, and excitation waveform on the specific core loss (SCL) and dynamic B-H loops. The results show that the ratio (R) of sine to square wave excitation specific core loss was always greater than unity for a given f and T and identical values of B(sub M). The values of R ranged from 1.07 to 1.34. The classical theory of core loss separation into a hysteresis and eddy current loss component was used to theoretically determine the lower and upper bounds on R, against which the experimental R-values were compared. The experimental R-values were also used to make a comparison of the core loss of a sine and square wave voltage driven transformer.

  6. Yersinia effector YopO uses actin as bait to phosphorylate proteins that regulate actin polymerization

    PubMed Central

    Lee, Wei Lin; Grimes, Jonathan M; Robinson, Robert C

    2016-01-01

    Pathogenic Yersinia species evade host immune systems through the injection of Yersinia outer proteins (Yops) into phagocytic cells. One Yop, YopO, also known as YpkA, induces actin-filament disruption, impairing phagocytosis. Here we describe the X-ray structure of Yersinia enterocolitica YopO in complex with actin, which reveals that YopO binds to an actin monomer in a manner that blocks polymerization yet allows the bound actin to interact with host actin-regulating proteins. SILAC-MS and biochemical analyses confirm that actin-polymerization regulators such as VASP, EVL, WASP, gelsolin and the formin diaphanous 1 are directly sequestered and phosphorylated by YopO through formation of ternary complexes with actin. This leads to a model in which YopO at the membrane sequesters actin from polymerization while using the bound actin as bait to recruit, phosphorylate and misregulate host actin-regulating proteins to disrupt phagocytosis. PMID:25664724

  7. Calcium storage and release properties of F-actin: evidence for the involvement of F-actin in cellular calcium signaling.

    PubMed

    Lange, K; Brandt, U

    1996-10-21

    Preceding studies have shown that the bulk of the ATP-dependent, inositol 1,4,5-trisphosphate (IP3)-sensitive Ca2+ store of hamster insulinoma (HIT) cells is located in microvilli on the cell surface. Similar results were obtained with isolated rat hepatocytes. Moreover, in vesicles of microvillar origin, passive fluxes of Ca2+, ATP, and IP3 occur through cation and anion channels, respectively, suggesting that Ca2+ storage is due to ATP-dependent Ca2+ binding to an intravesicular component. Here we demonstrate that F-actin may be a possible candidate for this function. ATP-actin monomers bind Ca2+ with high affinity (Kd = 2-8 nM) to their divalent cation binding sites. Polymerization of actin monomers decreases the rate constant for divalent cation exchange at this binding site by more than 3 orders of magnitude rendering bound cations nearly unavailable. F-actin-bound Ca2+ can be released by depolymerization and dissociation from Ca(2+)-ADP-actin monomers (Kd = 375 nM). We now provide additional evidence for the possible involvement of actin in Ca2+ storage. (1) Preincubation of surface-derived Ca(2+)-storing vesicles from HIT cells with the F-actin stabilizer, phalloidin, strongly inhibited ATP-dependent Ca2+ uptake, reducing the IP3-sensitive Ca2+ pool by 70%. Phalloidin, when added after the loading process, affected neither the amount of stored Ca2+ nor IP3 action on the store. (2) F-actin polymerized in the presence of Mg2+ in nominally Ca(2+)-free buffer still contained about half of the high affinity sites occupied with Ca2+ (Mg/Ca-F-actin). (3) Using the fura-2 technique, we found that in the presence of ATP, Mg/Ca-F-actin incorporated free Ca2+ at a relatively low rate. Short pulses of ultrasound (3-10 s) strongly accelerated Ca2+ uptake, decreasing free Ca2+ from 500 nM to below 100 nM. (4) In the presence of physiological levels of Mg2+ (0.5 mM), sonication liberated large amounts of Ca2+ from Mg/Ca-F-actin. (5) Ca-F-actin released bound Ca2+ at a very

  8. Actin Interacting Protein1 and Actin Depolymerizing Factor Drive Rapid Actin Dynamics in Physcomitrella patens[W

    PubMed Central

    Augustine, Robert C.; Pattavina, Kelli A.; Tüzel, Erkan; Vidali, Luis; Bezanilla, Magdalena

    2011-01-01

    The remodeling of actin networks is required for a variety of cellular processes in eukaryotes. In plants, several actin binding proteins have been implicated in remodeling cortical actin filaments (F-actin). However, the extent to which these proteins support F-actin dynamics in planta has not been tested. Using reverse genetics, complementation analyses, and cell biological approaches, we assessed the in vivo function of two actin turnover proteins: actin interacting protein1 (AIP1) and actin depolymerizing factor (ADF). We report that AIP1 is a single-copy gene in the moss Physcomitrella patens. AIP1 knockout plants are viable but have reduced expansion of tip-growing cells. AIP1 is diffusely cytosolic and functions in a common genetic pathway with ADF to promote tip growth. Specifically, ADF can partially compensate for loss of AIP1, and AIP1 requires ADF for function. Consistent with a role in actin remodeling, AIP1 knockout lines accumulate F-actin bundles, have fewer dynamic ends, and have reduced severing frequency. Importantly, we demonstrate that AIP1 promotes and ADF is essential for cortical F-actin dynamics. PMID:22003077

  9. Computational Tension Mapping of Adherent Cells Based on Actin Imaging

    PubMed Central

    Manifacier, Ian; Milan, Jean-Louis; Jeanneau, Charlotte; Chmilewsky, Fanny; Chabrand, Patrick; About, Imad

    2016-01-01

    Forces transiting through the cytoskeleton are known to play a role in adherent cell activity. Up to now few approaches haves been able to determine theses intracellular forces. We thus developed a computational mechanical model based on a reconstruction of the cytoskeleton of an adherent cell from fluorescence staining of the actin network and focal adhesions (FA). Our custom made algorithm converted the 2D image of an actin network into a map of contractile interactions inside a 2D node grid, each node representing a group of pixels. We assumed that actin filaments observed under fluorescence microscopy, appear brighter when thicker, we thus presumed that nodes corresponding to pixels with higher actin density were linked by stiffer interactions. This enabled us to create a system of heterogeneous interactions which represent the spatial organization of the contractile actin network. The contractility of this interaction system was then adapted to match the level of force the cell truly exerted on focal adhesions; forces on focal adhesions were estimated from their vinculin expressed size. This enabled the model to compute consistent mechanical forces transiting throughout the cell. After computation, we applied a graphical approach on the original actin image, which enabled us to calculate tension forces throughout the cell, or in a particular region or even in single stress fibers. It also enabled us to study different scenarios which may indicate the mechanical role of other cytoskeletal components such as microtubules. For instance, our results stated that the ratio between intra and extra cellular compression is inversely proportional to intracellular tension. PMID:26812601

  10. Computational Tension Mapping of Adherent Cells Based on Actin Imaging.

    PubMed

    Manifacier, Ian; Milan, Jean-Louis; Jeanneau, Charlotte; Chmilewsky, Fanny; Chabrand, Patrick; About, Imad

    2016-01-01

    Forces transiting through the cytoskeleton are known to play a role in adherent cell activity. Up to now few approaches haves been able to determine theses intracellular forces. We thus developed a computational mechanical model based on a reconstruction of the cytoskeleton of an adherent cell from fluorescence staining of the actin network and focal adhesions (FA). Our custom made algorithm converted the 2D image of an actin network into a map of contractile interactions inside a 2D node grid, each node representing a group of pixels. We assumed that actin filaments observed under fluorescence microscopy, appear brighter when thicker, we thus presumed that nodes corresponding to pixels with higher actin density were linked by stiffer interactions. This enabled us to create a system of heterogeneous interactions which represent the spatial organization of the contractile actin network. The contractility of this interaction system was then adapted to match the level of force the cell truly exerted on focal adhesions; forces on focal adhesions were estimated from their vinculin expressed size. This enabled the model to compute consistent mechanical forces transiting throughout the cell. After computation, we applied a graphical approach on the original actin image, which enabled us to calculate tension forces throughout the cell, or in a particular region or even in single stress fibers. It also enabled us to study different scenarios which may indicate the mechanical role of other cytoskeletal components such as microtubules. For instance, our results stated that the ratio between intra and extra cellular compression is inversely proportional to intracellular tension. PMID:26812601

  11. Mechanism of Actin-Based Motility

    NASA Astrophysics Data System (ADS)

    Pantaloni, Dominique; Le Clainche, Christophe; Carlier, Marie-France

    2001-05-01

    Spatially controlled polymerization of actin is at the origin of cell motility and is responsible for the formation of cellular protrusions like lamellipodia. The pathogens Listeria monocytogenes and Shigella flexneri, which undergo actin-based propulsion, are acknowledged models of the leading edge of lamellipodia. Actin-based motility of the bacteria or of functionalized microspheres can be reconstituted in vitro from only five pure proteins. Movement results from the regulated site-directed treadmilling of actin filaments, consistent with observations of actin dynamics in living motile cells and with the biochemical properties of the components of the synthetic motility medium.

  12. Orientational order of the lamellipodial actin network as demonstrated in living motile cells.

    PubMed

    Verkhovsky, Alexander B; Chaga, Oleg Y; Schaub, Sébastien; Svitkina, Tatyana M; Meister, Jean-Jacques; Borisy, Gary G

    2003-11-01

    Lamellipodia of crawling cells represent both the motor for cell advance and the primary building site for the actin cytoskeleton. The organization of actin in the lamellipodium reflects actin dynamics and is of critical importance for the mechanism of cell motility. In previous structural studies, the lamellipodial actin network was analyzed primarily by electron microscopy (EM). An understanding of lamellipodial organization would benefit significantly if the EM data were complemented and put into a kinetic context by establishing correspondence with structural features observable at the light microscopic level in living cells. Here, we use an enhanced phase contrast microscopy technique to visualize an apparent long-range diagonal actin meshwork in the advancing lamellipodia of living cells. Visualization of this meshwork permitted a correlative light and electron microscopic approach that validated the underlying organization of lamellipodia. The linear features in the light microscopic meshwork corresponded to regions of greater actin filament density. Orientation of features was analyzed quantitatively and compared with the orientation of actin filaments at the EM level. We infer that the light microscopic meshwork reflects the orientational order of actin filaments which, in turn, is related to their branching angle. PMID:13679520

  13. Orientational Order of the Lamellipodial Actin Network as Demonstrated in Living Motile CellsV⃞

    PubMed Central

    Verkhovsky, Alexander B.; Chaga, Oleg Y.; Schaub, Sébastien; Svitkina, Tatyana M.; Meister, Jean-Jacques; Borisy, Gary G.

    2003-01-01

    Lamellipodia of crawling cells represent both the motor for cell advance and the primary building site for the actin cytoskeleton. The organization of actin in the lamellipodium reflects actin dynamics and is of critical importance for the mechanism of cell motility. In previous structural studies, the lamellipodial actin network was analyzed primarily by electron microscopy (EM). An understanding of lamellipodial organization would benefit significantly if the EM data were complemented and put into a kinetic context by establishing correspondence with structural features observable at the light microscopic level in living cells. Here, we use an enhanced phase contrast microscopy technique to visualize an apparent long-range diagonal actin meshwork in the advancing lamellipodia of living cells. Visualization of this meshwork permitted a correlative light and electron microscopic approach that validated the underlying organization of lamellipodia. The linear features in the light microscopic meshwork corresponded to regions of greater actin filament density. Orientation of features was analyzed quantitatively and compared with the orientation of actin filaments at the EM level. We infer that the light microscopic meshwork reflects the orientational order of actin filaments which, in turn, is related to their branching angle. PMID:13679520

  14. Quantifying actin wave modulation on periodic topography

    NASA Astrophysics Data System (ADS)

    Guven, Can; Driscoll, Meghan; Sun, Xiaoyu; Parker, Joshua; Fourkas, John; Carlsson, Anders; Losert, Wolfgang

    2014-03-01

    Actin is the essential builder of the cell cytoskeleton, whose dynamics are responsible for generating the necessary forces for the formation of protrusions. By exposing amoeboid cells to periodic topographical cues, we show that actin can be directionally guided via inducing preferential polymerization waves. To quantify the dynamics of these actin waves and their interaction with the substrate, we modify a technique from computer vision called ``optical flow.'' We obtain vectors that represent the apparent actin flow and cluster these vectors to obtain patches of newly polymerized actin, which represent actin waves. Using this technique, we compare experimental results, including speed distribution of waves and distance from the wave centroid to the closest ridge, with actin polymerization simulations. We hypothesize the modulation of the activity of nucleation promotion factors on ridges (elevated regions of the surface) as a potential mechanism for the wave-substrate coupling. Funded by NIH grant R01GM085574.

  15. Arabidopsis Microtubule-Destabilizing Protein 25 Functions in Pollen Tube Growth by Severing Actin Filaments[W

    PubMed Central

    Qin, Tao; Liu, Xiaomin; Li, Jiejie; Sun, Jingbo; Song, Leina; Mao, Tonglin

    2014-01-01

    The formation of distinct actin filament arrays in the subapical region of pollen tubes is crucial for pollen tube growth. However, the molecular mechanisms underlying the organization and dynamics of the actin filaments in this region remain to be determined. This study shows that Arabidopsis thaliana MICROTUBULE-DESTABILIZING PROTEIN25 (MDP25) has the actin filament–severing activity of an actin binding protein. This protein negatively regulated pollen tube growth by modulating the organization and dynamics of actin filaments in the subapical region of pollen tubes. MDP25 loss of function resulted in enhanced pollen tube elongation and inefficient fertilization. MDP25 bound directly to actin filaments and severed individual actin filaments, in a manner that was dramatically enhanced by Ca2+, in vitro. Analysis of a mutant that bears a point mutation at the Ca2+ binding sites demonstrated that the subcellular localization of MDP25 was determined by cytosolic Ca2+ level in the subapical region of pollen tubes, where MDP25 was disassociated from the plasma membrane and moved into the cytosol. Time-lapse analysis showed that the F-actin-severing frequency significantly decreased and a high density of actin filaments was observed in the subapical region of mdp25-1 pollen tubes. This study reveals a mechanism whereby calcium enhances the actin filament–severing activity of MDP25 in the subapical region of pollen tubes to modulate pollen tube growth. PMID:24424096

  16. Reactive oxygen species (ROS)-induced actin glutathionylation controls actin dynamics in neutrophils

    PubMed Central

    Sakai, Jiro; Li, Jingyu; Subramanian, Kulandayan K.; Mondal, Subhanjan; Bajrami, Besnik; Hattori, Hidenori; Jia, Yonghui; Dickinson, Bryan C.; Zhong, Jia; Ye, Keqiang; Chang, Christopher J; Ho, Ye-Shih; Zhou, Jun; Luo, Hongbo R.

    2012-01-01

    Summary The regulation of actin dynamics is pivotal for cellular processes such as cell adhesion, migration, and phagocytosis, and thus is crucial for neutrophils to fulfill their roles in innate immunity. Many factors have been implicated in signal-induced actin polymerization, however the essential nature of the potential negative modulators are still poorly understood. Here we report that NADPH oxidase-dependent physiologically generated reactive oxygen species (ROS) negatively regulate actin polymerization in stimulated neutrophils via driving reversible actin glutathionylation. Disruption of glutaredoxin 1 (Grx1), an enzyme that catalyzes actin deglutathionylation, increased actin glutathionylation, attenuated actin polymerization, and consequently impaired neutrophil polarization, chemotaxis, adhesion, and phagocytosis. Consistently, Grx1-deficient murine neutrophils showed impaired in vivo recruitment to sites of inflammation and reduced bactericidal capability. Together, these results present a physiological role for glutaredoxin and ROS- induced reversible actin glutathionylation in regulation of actin dynamics in neutrophils. PMID:23159440

  17. Model for adhesion clutch explains biphasic relationship between actin flow and traction at the cell leading edge

    NASA Astrophysics Data System (ADS)

    Craig, Erin M.; Stricker, Jonathan; Gardel, Margaret; Mogilner, Alex

    2015-05-01

    Cell motility relies on the continuous reorganization of a dynamic actin-myosin-adhesion network at the leading edge of the cell, in order to generate protrusion at the leading edge and traction between the cell and its external environment. We analyze experimentally measured spatial distributions of actin flow, traction force, myosin density, and adhesion density in control and pharmacologically perturbed epithelial cells in order to develop a mechanical model of the actin-adhesion-myosin self-organization at the leading edge. A model in which the F-actin network is treated as a viscous gel, and adhesion clutch engagement is strengthened by myosin but weakened by actin flow, can explain the measured molecular distributions and correctly predict the spatial distributions of the actin flow and traction stress. We test the model by comparing its predictions with measurements of the actin flow and traction stress in cells with fast and slow actin polymerization rates. The model predicts how the location of the lamellipodium-lamellum boundary depends on the actin viscosity and adhesion strength. The model further predicts that the location of the lamellipodium-lamellum boundary is not very sensitive to the level of myosin contraction.

  18. Hysteretic Dependence of Magnetic Flux Density on Primary AC Current in Flat-Type Inductive Fault Current Limiter with YBCO Thin Film Discs

    NASA Astrophysics Data System (ADS)

    Harada, Masayuki; Yokomizu, Yasunobu; Matsumura, Toshiro

    2014-05-01

    This paper focuses on a flat-type inductive superconducting FCL (FIS-FCL) consisting of a pancake coil and a YBCO thin layer disc. AC current injection experiments and magnetic field analysis were carried out for two kinds of FIS-FCL, single-disc model and double-discs model. In the former, the pancake coil was putted on the YBCO disc. In the latter, the pancake coil was sandwiched with two YBCO discs. The double-discs model cancels out the magnetic flux density more effectively than the single-disc model. In the double-discs model, the superconducting state period is longer than in the single-disc model. Thus, it may be concluded that the double-discs model is considered to be suitable for FIS-FCL.

  19. Continuum Contrast as a function of Magnetic Flux Density and Disk Position: Results from a full Solar Cycle of SOHO/MDI data

    NASA Astrophysics Data System (ADS)

    Soto, K.; Basri, G.; Ramos-Stierle, F.; Lewis, T.; Reiners, A.; Berger, T.

    2006-12-01

    The key to understanding total solar irradiance variability is to understand the mechanisms by continuum contrast is effected by the distribution of magnetic flux across the surface of the Sun. The Michelson Doppler Imager (MDI) instrument on the Solar and Heliospheric Observatory (SOHO) satellite has measured full-disk Stokes-V magnetograms along with cotemporal continuum images throughout Solar Cycle 23. We present the results of an examination of the relationship between corrected MDI continuum images with their cotemporal corrected MDI magnetograms. By separating the series of images into bins of Magnetic Flux Density (MFD) and limb angle, we perform a non-linear least squares fit to the data to produce a 3rd-order polynomial function of continuum contrast vs. MFD and disk position. Using the obtained function we can accurately predict continuum contrast at any given disk position as a function of MFD measured in a given magnetogram. The resulting predictions can be used to make simulated continuum images suitable for bolometric correction and input into total irradiance models. This research was partially supported by LWS TR&T Grant NNG05GK46G

  20. Upper critical field, critical current density and thermally activated flux flow in CaFFe0.9Co0.1As superconductor

    NASA Astrophysics Data System (ADS)

    Shekhar, Chandra; Srivastava, Amit; Kumar, Pramod; Srivastava, Pankaj; Srivastava, O. N.

    2012-04-01

    In this paper, we report the synthesis, structure, transition temperature, upper critical field, critical current density and thermally activated flux flow in the CaFFe0.9Co0.1As superconductor. Superconductivity arises at 23 K by Co substitution at the site of Fe atoms and the upper critical field is estimated as 102 T using the Werthamer-Helfand-Hohenberg formula. The flux-flow activation energy (U0/kB) varies from 3230 K and 4190 K in a field of 9 T and 1 T, respectively. At 2 K, the Jc is found to be approximately 4 × 103 A cm-2 and 0.3 × 103 A cm-2 in zero and 6 T field, respectively. Transmission electron microscopy analysis shows an amorphous region surrounding most of the grains which is likely to be present in the form of amorphous and weak link grain boundaries in this compound. It seems that most of the current is hindered by mis-aligned grains, amorphous grain boundaries and impurities, which are invariably found between the grains. The presence of the weakly linked granules and their weakly pinned intergranular Josephson vortices are responsible for both low Jc and the Arrhenius temperature dependence of resistivity.

  1. Mesoscopic model of actin-based propulsion.

    PubMed

    Zhu, Jie; Mogilner, Alex

    2012-01-01

    Two theoretical models dominate current understanding of actin-based propulsion: microscopic polymerization ratchet model predicts that growing and writhing actin filaments generate forces and movements, while macroscopic elastic propulsion model suggests that deformation and stress of growing actin gel are responsible for the propulsion. We examine both experimentally and computationally the 2D movement of ellipsoidal beads propelled by actin tails and show that neither of the two models can explain the observed bistability of the orientation of the beads. To explain the data, we develop a 2D hybrid mesoscopic model by reconciling these two models such that individual actin filaments undergoing nucleation, elongation, attachment, detachment and capping are embedded into the boundary of a node-spring viscoelastic network representing the macroscopic actin gel. Stochastic simulations of this 'in silico' actin network show that the combined effects of the macroscopic elastic deformation and microscopic ratchets can explain the observed bistable orientation of the actin-propelled ellipsoidal beads. To test the theory further, we analyze observed distribution of the curvatures of the trajectories and show that the hybrid model's predictions fit the data. Finally, we demonstrate that the model can explain both concave-up and concave-down force-velocity relations for growing actin networks depending on the characteristic time scale and network recoil. To summarize, we propose that both microscopic polymerization ratchets and macroscopic stresses of the deformable actin network are responsible for the force and movement generation. PMID:23133366

  2. Affinity chromatography of immobilized actin and myosin.

    PubMed Central

    Bottomley, R C; Trayer, I P

    1975-01-01

    Actin and myosin were immobilized by coupling them to agarose matrices. Both immobilized G-actin and immobilized myosin retain most of the properties of the proteins in free solution and are reliable over long periods of time. Sepharose-F-actin, under the conditions used in this study, has proved unstable and variable in its properties. Sepharose-G-actin columns were used to bind heavy meromyosin and myosin subfragment 1 specifically and reversibly. The interaction involved is sensitive to variation in ionic strength, such that myosin itself is not retained by the columns at the high salt concentration required for its complete solubilization. Myosin, rendered soluble at low ionic strength by polyalanylation, will interact successfully with the immobilized actin. The latter can distinguish between active and inactive fractions of the proteolytic and polyalanyl myosin derivatives, and was used in the preparation of these molecules. The complexes formed between the myosin derivatives and Sepharose-G-actin can be dissociated by low concentrations of ATP, ADP and pyrophosphate in both the presence and the absence of Mg2+. The G-actin columns were used to evaluate the results of chemical modifications of myosin subfragments on their interactions with actin. F-Actin in free solution is bound specifically and reversibly to columns of insolubilized myosin. Thus, with elution by either ATP or pyrophosphate, actin has been purified in one step from extracts of acetone-dried muscle powder. PMID:241335

  3. The interaction of vinculin with actin.

    PubMed

    Golji, Javad; Mofrad, Mohammad R K

    2013-04-01

    Vinculin can interact with F-actin both in recruitment of actin filaments to the growing focal adhesions and also in capping of actin filaments to regulate actin dynamics. Using molecular dynamics, both interactions are simulated using different vinculin conformations. Vinculin is simulated either with only its vinculin tail domain (Vt), with all residues in its closed conformation, with all residues in an open I conformation, and with all residues in an open II conformation. The open I conformation results from movement of domain 1 away from Vt; the open II conformation results from complete dissociation of Vt from the vinculin head domains. Simulation of vinculin binding along the actin filament showed that Vt alone can bind along the actin filaments, that vinculin in its closed conformation cannot bind along the actin filaments, and that vinculin in its open I conformation can bind along the actin filaments. The simulations confirm that movement of domain 1 away from Vt in formation of vinculin 1 is sufficient for allowing Vt to bind along the actin filament. Simulation of Vt capping actin filaments probe six possible bound structures and suggest that vinculin would cap actin filaments by interacting with both S1 and S3 of the barbed-end, using the surface of Vt normally occluded by D4 and nearby vinculin head domain residues. Simulation of D4 separation from Vt after D1 separation formed the open II conformation. Binding of open II vinculin to the barbed-end suggests this conformation allows for vinculin capping. Three binding sites on F-actin are suggested as regions that could link to vinculin. Vinculin is suggested to function as a variable switch at the focal adhesions. The conformation of vinculin and the precise F-actin binding conformation is dependent on the level of mechanical load on the focal adhesion. PMID:23633939

  4. An easy-to-use single-molecule speckle microscopy enabling nanometer-scale flow and wide-range lifetime measurement of cellular actin filaments.

    PubMed

    Yamashiro, Sawako; Mizuno, Hiroaki; Watanabe, Naoki

    2015-01-01

    Single-molecule speckle (SiMS) microscopy has been a powerful method to analyze actin dynamics in live cells by tracking single molecule of fluorescently labeled actin. Recently we developed a new SiMS method, which is easy-to-use for inexperienced researchers and achieves high spatiotemporal resolution. In this method, actin labeled with fluorescent DyLight dye on lysines is employed as a probe. Electroporation-mediated delivery of DyLight-actin (DL-actin) into cells enables to label cells with 100% efficiency at the optimal density. DL-actin labels cellular actin filaments including formin-based structures with improved photostability and brightness compared to green fluorescent protein-actin. These favorable properties of DL-actin extend time window of the SiMS analysis. Furthermore, the new SiMS method enables nanometer-scale displacement analysis with a low localization error of ±8-8.5 nm. With these advantages, our new SiMS microscopy method will help researchers to investigate various actin remodeling processes. In this chapter, we introduce the methods for preparation of DL-actin probes, electroporation to deliver DL-actin, the SiMS imaging and data analysis. PMID:25640423

  5. Mechanosensitive kinetic preference of actin-binding protein to actin filament

    NASA Astrophysics Data System (ADS)

    Inoue, Yasuhiro; Adachi, Taiji

    2016-04-01

    The kinetic preference of actin-binding proteins to actin filaments is altered by external forces on the filament. Such an altered kinetic preference is largely responsible for remodeling the actin cytoskeletal structure in response to intracellular forces. During remodeling, actin-binding proteins and actin filaments interact under isothermal conditions, because the cells are homeostatic. In such a temperature homeostatic state, we can rigorously and thermodynamically link the chemical potential of actin-binding proteins to stresses on the actin filaments. From this relationship, we can construct a physical model that explains the force-dependent kinetic preference of actin-binding proteins to actin filaments. To confirm the model, we have analyzed the mechanosensitive alternation of the kinetic preference of Arp2/3 and cofilin to actin filaments. We show that this model captures the qualitative responses of these actin-binding proteins to the forces, as observed experimentally. Moreover, our theoretical results demonstrate that, depending on the structural parameters of the binding region, actin-binding proteins can show different kinetic responses even to the same mechanical signal tension, in which the double-helix nature of the actin filament also plays a critical role in a stretch-twist coupling of the filament.

  6. Balloon-borne measurements of the ultraviolet flux in the Arctic stratosphere during winter

    NASA Technical Reports Server (NTRS)

    Schiller, Cornelius; Mueller, Martin; Klein, Erich; Schmidt, Ulrich; Roeth, Ernst-Peter

    1994-01-01

    Filter radiometers sensitive from 280 to 320 nm and from 280 to 400 nm, respectively, were used for measurements of the actinic flux in the stratosphere. Since the instruments are calibrated for absolute spectral sensitivity the data can be compared with model calculations of the actinic flux. Data were obtained during seven balloon flights during the European Arctic Stratospheric Ozone Experiment (EASOE).

  7. Elasticity, adhesion and actin based propulsion

    NASA Astrophysics Data System (ADS)

    Gopinathan, Ajay

    2006-03-01

    When a cells crawls, its shape re-organizes via polymerization and depolymerization of actin filaments. The growing ends of the filaments are oriented towards the outside of the cell, and their polymerization pushes the cell membrane forwards. The same mechanism comes into play when the bacterial pathogen Listeria monocytogenes infects a cell. The bacterium hijacks the host cell's actin machinery to create an actin network (the actin comet tail) that propels the bacterium through cells and into neighboring cells. We propose a mechanism for how polymerization gives rise to motility that incorporates the effects of inhomogeneous polymerization. We treat the actin comet tail as an elastic continuum tethered to the rear of the bacterium. The interplay of polymerization and tethering gives rise to inhomogeneous stresses calculated with a finite element analysis. We quantitatively reproduce many distinctive features of actin propulsion that have been observed experimentally, including stepped motion, hopping, tail shape and the propulsion of flat surfaces.

  8. Functional interdependence between septin and actin cytoskeleton

    PubMed Central

    Schmidt, Katja; Nichols, Benjamin J

    2004-01-01

    Background Septin2 is a member of a highly conserved GTPase family found in fungi and animals. Septins have been implicated in a diversity of cellular processes including cytokinesis, formation of diffusion barriers and vesicle trafficking. Septin2 partially co-localises with actin bundles in mammalian interphase cells and Septin2-filamentmorphology depends upon an intact actin cytoskeleton. How this interaction is regulated is not known. Moreover, evidence that Septin2 is remodelled or redistributed in response to other changes in actin organisation is lacking. Results Septin2 filaments are associated with actin fibres, but Septin2 is not associated with actin at the leading edge of moving cells or in ruffles where actin is highly dynamic. Rather, Septin2 is spatially segregated from these active areas and forms O- and C-shaped structures, similar to those previously observed after latrunculin treatment. FRAP experiments showed that all assemblies formed by Septin2 are highly dynamic with a constant exchange of Septin2 in and out of these structures, and that this property is independent of actin. A combination of RNAi experiments and expression of truncated forms of Septin2 showed that Septin2 plays a significant role in stabilising or maintaining actin bundles. Conclusion We show that Septin2 can form dynamic structures with differing morphologies in living cells, and that these morphologies are dependent on the functional state of the actin cytoskeleton. Our data provide a link between the different morphological states of Septin2 and functions of Septin2 in actin-dynamics, and are consistent with the model proposed by Kinoshita and colleagues, that Septin2 filaments play a role in stabilisation of actin stress fibres thus preventing actin turnover. PMID:15541171

  9. Rho, nuclear actin, and actin-binding proteins in the regulation of transcription and gene expression

    PubMed Central

    Rajakylä, Eeva Kaisa; Vartiainen, Maria K

    2014-01-01

    Actin cytoskeleton is one of the main targets of Rho GTPases, which act as molecular switches on many signaling pathways. During the past decade, actin has emerged as an important regulator of gene expression. Nuclear actin plays a key role in transcription, chromatin remodeling, and pre-mRNA processing. In addition, the “status” of the actin cytoskeleton is used as a signaling intermediate by at least the MKL1-SRF and Hippo-pathways, which culminate in the transcriptional regulation of cytoskeletal and growth-promoting genes, respectively. Rho GTPases may therefore regulate gene expression by controlling either cytoplasmic or nuclear actin dynamics. Although the regulation of nuclear actin polymerization is still poorly understood, many actin-binding proteins, which are downstream effectors of Rho, are found in the nuclear compartment. In this review, we discuss the possible mechanisms and key proteins that may mediate the transcriptional regulation by Rho GTPases through actin. PMID:24603113

  10. Rho, nuclear actin, and actin-binding proteins in the regulation of transcription and gene expression.

    PubMed

    Rajakylä, Eeva Kaisa; Vartiainen, Maria K

    2014-01-01

    Actin cytoskeleton is one of the main targets of Rho GTPases, which act as molecular switches on many signaling pathways. During the past decade, actin has emerged as an important regulator of gene expression. Nuclear actin plays a key role in transcription, chromatin remodeling, and pre-mRNA processing. In addition, the "status" of the actin cytoskeleton is used as a signaling intermediate by at least the MKL1-SRF and Hippo-pathways, which culminate in the transcriptional regulation of cytoskeletal and growth-promoting genes, respectively. Rho GTPases may therefore regulate gene expression by controlling either cytoplasmic or nuclear actin dynamics. Although the regulation of nuclear actin polymerization is still poorly understood, many actin-binding proteins, which are downstream effectors of Rho, are found in the nuclear compartment. In this review, we discuss the possible mechanisms and key proteins that may mediate the transcriptional regulation by Rho GTPases through actin. PMID:24603113

  11. Calcium control of Saccharomyces cerevisiae actin assembly.

    PubMed Central

    Greer, C; Schekman, R

    1982-01-01

    Low levels of Ca2+ dramatically influence the polymerization of Saccharomyces cerevisiae actin in KCl. The apparent critical concentration for polymerization (C infinity) increases eightfold in the presence of 0.1 mM Ca2+. This effect is rapidly reversed by the addition of ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid or of 0.1 mM Mg2+. Furthermore, the addition of Ca2+ to polymerized actin causes a reversible increase in the apparent C infinity. In the presence of Ca2+, at actin concentrations below the apparent C infinity, particles of 15 to 50 nm in diameter are seen instead of filaments. These particles are separated from soluble actin when Ca2+-treated filamentous actin is sedimented at high speed; both the soluble and particulate fractions retain Ca2+-sensitive polymerization. The Ca2+ effect is S. cerevisiae actin-specific: the C infinity for rabbit muscle actin is not affected by the presence of Ca2+ and S. cerevisiae actin. Ca2+ may act directly on S. cerevisiae actin to control the assembly state in vivo. Images PMID:6757718

  12. Architecture and Connectivity Govern Actin Network Contractility.

    PubMed

    Ennomani, Hajer; Letort, Gaëlle; Guérin, Christophe; Martiel, Jean-Louis; Cao, Wenxiang; Nédélec, François; De La Cruz, Enrique M; Théry, Manuel; Blanchoin, Laurent

    2016-03-01

    Actomyosin contractility plays a central role in a wide range of cellular processes, including the establishment of cell polarity, cell migration, tissue integrity, and morphogenesis during development. The contractile response is variable and depends on actomyosin network architecture and biochemical composition. To determine how this coupling regulates actomyosin-driven contraction, we used a micropatterning method that enables the spatial control of actin assembly. We generated a variety of actin templates and measured how defined actin structures respond to myosin-induced forces. We found that the same actin filament crosslinkers either enhance or inhibit the contractility of a network, depending on the organization of actin within the network. Numerical simulations unified the roles of actin filament branching and crosslinking during actomyosin contraction. Specifically, we introduce the concept of "network connectivity" and show that the contractions of distinct actin architectures are described by the same master curve when considering their degree of connectivity. This makes it possible to predict the dynamic response of defined actin structures to transient changes in connectivity. We propose that, depending on the connectivity and the architecture, network contraction is dominated by either sarcomeric-like or buckling mechanisms. More generally, this study reveals how actin network contractility depends on its architecture under a defined set of biochemical conditions. PMID:26898468

  13. Dynamic actin gene family evolution in primates.

    PubMed

    Zhu, Liucun; Zhang, Ying; Hu, Yijun; Wen, Tieqiao; Wang, Qiang

    2013-01-01

    Actin is one of the most highly conserved proteins and plays crucial roles in many vital cellular functions. In most eukaryotes, it is encoded by a multigene family. Although the actin gene family has been studied a lot, few investigators focus on the comparison of actin gene family in relative species. Here, the purpose of our study is to systematically investigate characteristics and evolutionary pattern of actin gene family in primates. We identified 233 actin genes in human, chimpanzee, gorilla, orangutan, gibbon, rhesus monkey, and marmoset genomes. Phylogenetic analysis showed that actin genes in the seven species could be divided into two major types of clades: orthologous group versus complex group. Codon usages and gene expression patterns of actin gene copies were highly consistent among the groups because of basic functions needed by the organisms, but much diverged within species due to functional diversification. Besides, many great potential pseudogenes were found with incomplete open reading frames due to frameshifts or early stop codons. These results implied that actin gene family in primates went through "birth and death" model of evolution process. Under this model, actin genes experienced strong negative selection and increased the functional complexity by reproducing themselves. PMID:23841080

  14. Stochastic model of profilin-actin polymerization

    NASA Astrophysics Data System (ADS)

    Horan, Brandon; Vavylonis, Dimitrios

    A driving factor in cell motility and other processes that involve changes of cell shape is the rapid polymerization of actin subunits into long filaments. This process is regulated by profilin, a protein which binds to actin subunits and regulates elongation of actin filaments. Whether profilin stimulates polymerization by coupling to hydrolysis of ATP-bound actin is debated. Previous studies have proposed indirect coupling to ATP hydrolysis using rate equations, but did not include the effects of fluctuations that are important near the critical concentration. We developed stochastic simulations using the Gillespie algorithm to study single filament elongation at the barbed end in the presence of profilin. We used recently measured rate constants and estimated the rate of profilin binding to the barbed end such that detailed balance is satisfied. Fast phosphate release at the tip of the filament was accounted for. The elongation rate and length diffusivity as functions of profilin and actin concentration were calculated and used to extract the critical concentrations of free actin and of total actin. We show under what conditions profilin leads to an increase in the critical concentration of total actin but a decrease in the critical concentration of free actin.

  15. F-actin waves, actin cortex disassembly and focal exocytosis driven by actin-phosphoinositide positive feedback.

    PubMed

    Masters, Thomas A; Sheetz, Michael P; Gauthier, Nils C

    2016-04-01

    Actin polymerization is controlled by the phosphoinositide composition of the plasma membrane. However, the molecular mechanisms underlying the spatiotemporal regulation of actin network organization over extended length scales are still unclear. To observe phosphoinositide-dependent cytoskeletal dynamics we combined the model system of frustrated phagocytosis, total internal reflection microscopy and manipulation of the buffer tonicity. We found that macrophages interacting with IgG-coated glass substrates formed circular F-actin waves on their ventral surface enclosing a region of plasma membrane devoid of cortical actin. Plasma membrane free of actin cortex was strongly depleted of PI(4,5)P2 , but enriched in PI(3,4)P2 and displayed a fivefold increase in exocytosis. Wave formation could be promoted by application of a hypotonic shock. The actin waves were characteristic of a bistable wavefront at the boundary between the regions of membrane containing and lacking cortical actin. Phosphoinositide modifiers and RhoGTPase activities dramatically redistributed with respect to the wavefronts, which often exhibited spatial oscillations. Perturbation of either lipid or actin cytoskeleton-related pathways led to rapid loss of both the polarized lipid distribution and the wavefront. As waves travelled over the plasma membrane, wavefront actin was seen to rapidly polymerize and depolymerize at pre-existing clusters of FcγRIIA, coincident with rapid changes in lipid composition. Thus the potential of receptors to support rapid F-actin polymerization appears to depend acutely on the local concentrations of multiple lipid species. We propose that interdependence through positive feedback from the cytoskeleton to lipid modifiers leads to coordinated local cortex remodeling, focal exocytosis, and organizes extended actin networks. PMID:26915738

  16. Statistics of actin-propelled trajectories in noisy environments.

    PubMed

    Wen, Fu-Lai; Chen, Hsuan-Yi; Leung, Kwan-Tai

    2016-06-01

    Actin polymerization is ubiquitously utilized to power the locomotion of eukaryotic cells and pathogenic bacteria in living systems. Inevitably, actin polymerization and depolymerization proceed in a fluctuating environment that renders the locomotion stochastic. Previously, we have introduced a deterministic model that manages to reproduce actin-propelled trajectories in experiments, but not to address fluctuations around them. To remedy this, here we supplement the deterministic model with noise terms. It enables us to compute the effects of fluctuating actin density and forces on the trajectories. Specifically, the mean-squared displacement (MSD) of the trajectories is computed and found to show a super-ballistic scaling with an exponent 3 in the early stage, followed by a crossover to a normal, diffusive scaling of exponent 1 in the late stage. For open-end trajectories such as straights and S-shaped curves, the time of crossover matches the decay time of orientational order of the velocities along trajectories, suggesting that it is the spreading of velocities that leads to the crossover. We show that the super-ballistic scaling of MSD arises from the initial, linearly increasing correlation of velocities, before time translational symmetry is established. When the spreading of velocities reaches a steady state in the long-time limit, short-range correlation then yields a diffusive scaling in MSD. In contrast, close-loop trajectories like circles exhibit localized periodic motion, which inhibits spreading. The initial super-ballistic scaling of MSD arises from velocity correlation that both linearly increases and oscillates in time. Finally, we find that the above statistical features of the trajectories transcend the nature of noises, be it additive or multiplicative, and generalize to other self-propelled systems that are not necessarily actin based. PMID:27415296

  17. Statistics of actin-propelled trajectories in noisy environments

    NASA Astrophysics Data System (ADS)

    Wen, Fu-Lai; Chen, Hsuan-Yi; Leung, Kwan-tai

    2016-06-01

    Actin polymerization is ubiquitously utilized to power the locomotion of eukaryotic cells and pathogenic bacteria in living systems. Inevitably, actin polymerization and depolymerization proceed in a fluctuating environment that renders the locomotion stochastic. Previously, we have introduced a deterministic model that manages to reproduce actin-propelled trajectories in experiments, but not to address fluctuations around them. To remedy this, here we supplement the deterministic model with noise terms. It enables us to compute the effects of fluctuating actin density and forces on the trajectories. Specifically, the mean-squared displacement (MSD) of the trajectories is computed and found to show a super-ballistic scaling with an exponent 3 in the early stage, followed by a crossover to a normal, diffusive scaling of exponent 1 in the late stage. For open-end trajectories such as straights and S-shaped curves, the time of crossover matches the decay time of orientational order of the velocities along trajectories, suggesting that it is the spreading of velocities that leads to the crossover. We show that the super-ballistic scaling of MSD arises from the initial, linearly increasing correlation of velocities, before time translational symmetry is established. When the spreading of velocities reaches a steady state in the long-time limit, short-range correlation then yields a diffusive scaling in MSD. In contrast, close-loop trajectories like circles exhibit localized periodic motion, which inhibits spreading. The initial super-ballistic scaling of MSD arises from velocity correlation that both linearly increases and oscillates in time. Finally, we find that the above statistical features of the trajectories transcend the nature of noises, be it additive or multiplicative, and generalize to other self-propelled systems that are not necessarily actin based.

  18. Effects of F/G-actin ratio and actin turn-over rate on NADPH oxidase activity in microglia

    PubMed Central

    2010-01-01

    Background Most in vivo studies that have addressed the role of actin dynamics in NADPH oxidase function in phagocytes have used toxins to modulate the polymerization state of actin and mostly effects on actin has been evaluated by end point measurements of filamentous actin, which says little about actin dynamics, and without consideration for the subcellular distribution of the perturbed actin cytoskeleton. Results Here, we in addition to toxins use conditional expression of the major actin regulatory protein LIM kinase-1 (LIMK1), and shRNA knock-down of cofilin to modulate the cellular F/G-actin ratio in the Ra2 microglia cell line, and we use Fluorescence Recovery after Photobleaching (FRAP) in β-actin-YFP-transduced cells to obtain a dynamic measure of actin recovery rates (actin turn-over rates) in different F/G-actin states of the actin cytoskeleton. Our data demonstrate that stimulated NADPH oxidase function was severely impaired only at extreme actin recovery rates and F/G-actin ratios, and surprisingly, that any moderate changes of these parameters of the actin cytoskeleton invariably resulted in an increased NADPH oxidase activity. Conclusion moderate actin polymerization and depolymerization both increase the FMLP and PMA-stimulated NADPH oxidase activity of microglia, which is directly correlated with neither actin recovery rate nor F/G- actin ratio. Our results indicate that NADPH oxidase functions in an enhanced state of activity in stimulated phagocytes despite widely different states of the actin cytoskeleton. PMID:20825680

  19. Force of an actin spring

    NASA Astrophysics Data System (ADS)

    Shin, Jennifer; Mahadevan, L.; Matsudaira, Paul

    2003-03-01

    The acrosomal process of the horseshoe crab sperm is a novel mechanochemical molecular spring that converts its elastic stain energy to mechanical work upon the chemical activation by Ca2+. Twisted and bent, the initial state of the acrosomal bundle features a high degree of complexity in its structure and the energy is believed to be stored in the highly strained actin filaments as an elastic potential energy. When activated, the bundle relaxes from the coil of the highly twisted and bent filaments to its straight conformation at a mean velocity of 15um/s. The mean extension velocity increases dramatically from 3um/s to 27um/s when temperature of the medium is changed from 9.6C to 32C (respective viscosities of 1.25-0.75cp), yet it exhibits a very weak dependence on changes in the medium viscosity (1cp-33cp). These experiments suggest that the uncoiling of the actin spring should be limited not by the viscosity of the medium but by the unlatching events of involved proteins at a molecular level. Unlike the viscosity-limited processes, where force is directly related to the rate of the reaction, a direct measurement is required to obtain the spring force of the acrosomal process. The extending acrosomal bundle is forced to push against a barrier and its elastic buckling response is analyzed to measure the force generated during the uncoiling.

  20. Dynamics of active actin networks

    NASA Astrophysics Data System (ADS)

    Koehler, Simone

    2014-03-01

    Local mechanical and structural properties of a eukaryotic cell are determined by its cytoskeleton. To adapt to their environment, cells rely on constant self-organized rearrangement processes of their actin cytoskeleton. To shed light on the principles underlying these dynamic self-organization processes we investigate a minimal reconstituted active system consisting of actin filaments, crosslinking molecules and molecular motor filaments. Using quantitative fluorescence microscopy and image analysis, we show, that these minimal model systems exhibit a generic structure formation mechanism. The competition between force generation by molecular motors and the stabilization of the network by crosslinking proteins results in a highly dynamic reorganization process which is characterized by anomalous transport dynamics with a superdiffusive behavior also found in intracellular dynamics. In vitro, these dynamics are governed by chemical and physical parameters that alter the balance of motor and crosslinking proteins, such as pH. These findings can be expected to have broad implications in our understanding of cytoskeletal regulation in vivo.

  1. Actin-Regulator Feedback Interactions during Endocytosis.

    PubMed

    Wang, Xinxin; Galletta, Brian J; Cooper, John A; Carlsson, Anders E

    2016-03-29

    Endocytosis mediated by clathrin, a cellular process by which cells internalize membrane receptors and their extracellular ligands, is an important component of cell signaling regulation. Actin polymerization is involved in endocytosis in varying degrees depending on the cellular context. In yeast, clathrin-mediated endocytosis requires a pulse of polymerized actin and its regulators, which recruit and activate the Arp2/3 complex. In this article, we seek to identify the main protein-protein interactions that 1) cause actin and its regulators to appear in pulses, and 2) determine the effects of key mutations and drug treatments on actin and regulator assembly. We perform a joint modeling/experimental study of actin and regulator dynamics during endocytosis in the budding yeast Saccharomyces cerevisiae. We treat both a stochastic model that grows an explicit three-dimensional actin network, and a simpler two-variable Fitzhugh-Nagumo type model. The models include a negative-feedback interaction of F-actin onto the Arp2/3 regulators. Both models explain the pulse time courses and the effects of interventions on actin polymerization: the surprising increase in the peak F-actin count caused by reduced regulator branching activity, the increase in F-actin resulting from slowing of actin disassembly, and the increased Arp2/3 regulator lifetime resulting from latrunculin treatment. In addition, they predict that decreases in the regulator branching activity lead to increases in accumulation of regulators, and we confirmed this prediction with experiments on yeast harboring mutations in the Arp2/3 regulators, using quantitative fluorescence microscopy. Our experimental measurements suggest that the regulators act quasi-independently, in the sense that accumulation of a particular regulator is most strongly affected by mutations of that regulator, as opposed to the others. PMID:27028652

  2. Three-dimensional structure of the complex of actin and DNase I at 4.5 A resolution.

    PubMed Central

    Kabsch, W; Mannherz, H G; Suck, D

    1985-01-01

    The shape of an actin subunit has been derived from an improved 6 A map of the complex of rabbit skeletal muscle actin and bovine pancreatic DNase I obtained by X-ray crystallographic methods. The three-dimensional structure of DNase I determined independently at 2.5 A resolution was compared with the DNase I electron density in the actin:DNase map. The two structures are very similar at 6 A resolution thus leading to an unambiguous identification of actin as well as DNase I electron density. Furthermore the correct hand of the actin structure is determined from the DNase I atomic structure. The resolution of the actin structure was extended to 4.5 A by using a single heavy-atom derivative and the knowledge of the atomic coordinates of DNase I. The dimensions of an actin subunit are 67 A X 40 A X 37 A. It consists of a small and a large domain, the small domain containing the N terminus. Actin is an alpha,beta-protein with a beta-pleated sheet in each domain. These sheets are surrounded by several alpha-helices, comprising at least 40% of the structure. The phosphate peak of the adenine nucleotide is located between the two domains. The complex of actin and DNase I as found in solution (i.e., the actin:DNase I contacts which do not depend on crystal packing) was deduced from a comparison of monoclinic with orthorhombic crystals. Residues 44-46, 51, 52, 60-62 of DNase I are close to a loop region in the small domain of actin. At a distance of approximately 15 A there is a second contact in the large domain in which Glu13 of DNase I is involved. A possible binding region for myosin is discussed. Images Fig. 1. Fig. 2. Fig. 3. PMID:4065103

  3. Effect of alpha-actinin on actin structure. Actin ATPase activity.

    PubMed

    Singh, I; Goll, D E; Robson, R M

    1981-08-28

    Alpha-Actinin increases the ATPase activity of actin by up to 84%, depending un pH, divalent cations present and the added Mg2+: ATP ratio. Dithiothreitol decreases actin ATPase activity approx. 20% but does not reduce the ability of alpha-actinin to increase actin ATP activity. Increasing amounts of added alpha-actinin up to 1 mos alpha-actinin to 49 mol actin cause in increasing increment in actin ATPase activity, but adding alpha-actinin beyond 1 mol alpha-actinin to 49 mol actin elicits only small additional increments in activity. Actin ATPase activity ranges from approx 100 nmol Pi/mg actin per h (4.3 mol Pi/mol actin per h) at high levels (10 mM) of ATP in the presence of lower amounts (1 mM) of added mg2+ to approx. 12.5 nmol Pi/mg actin per h (0.52 mol Pi/mol actin per h) at high pH (8.5) or at low levels (0.5-1.0 mM) of ATP in the presence of higher amounts (10 mM) of added Mg2+ ATp uncomplexed with Mg2+ inhibits the ability of alpha-actinin to increase F-actin ATPase activity. Activities with different divalent cations showed that the actin ATPase in these studies, which was 1/100 as great as Mg2+-modified actomyosin ATPase activity, was not due to trace amounts of myosin contaminating the actin preparations. The results are consistent with the concept that alpha-actinin can alter the structure of actin monomers. PMID:6456018

  4. Synthetic peptides that cause F-actin bundling and block actin depolymerization

    DOEpatents

    Sederoff, Heike; Huber, Steven C; Larabell, Carolyn A

    2011-10-18

    Synthetic peptides derived from sucrose synthase, and having homology to actin and actin-related proteins, sharing a common motif, useful for causing acting bundling and preventing actin depolymerization. Peptides exhibiting the common motif are described, as well as specific synthetic peptides which caused bundled actin and inhibit actin depolymerization. These peptides can be useful for treating a subject suffering from a disease characterized by cells having neoplastic growth, for anti-cancer therapeutics, delivered to subjects solely, or concomitantly or sequentially with other known cancer therapeutics. These peptides can also be used for stabilizing microfilaments in living cells and inhibiting growth of cells.

  5. Extracellular signaling cues for nuclear actin polymerization.

    PubMed

    Plessner, Matthias; Grosse, Robert

    2015-01-01

    Contrary to cytoplasmic actin structures, the biological functions of nuclear actin filaments remain largely enigmatic. Recent progress in the field, however, has determined nuclear actin structures in somatic cells either under steady state conditions or in response to extracellular signaling cues. These actin structures differ in size and shape as well as in their temporal appearance and dynamics. Thus, a picture emerges that suggests that mammalian cells may have different pathways and mechanisms to assemble nuclear actin filaments. Apart from serum- or LPA-triggered nuclear actin polymerization, integrin activation by extracellular matrix interaction was recently implicated in nuclear actin polymerization through the linker of nucleoskeleton and cytoskeleton (LINC) complex. Some of these extracellular cues known so far appear to converge at the level of nuclear formin activity and subsequent regulation of myocardin-related transcription factors. Nevertheless, as the precise signaling events are as yet unknown, the regulation of nuclear actin polymerization may be of significant importance for different cellular functions as well as disease conditions caused by altered nuclear dynamics and architecture. PMID:26059398

  6. Profilin connects actin assembly with microtubule dynamics.

    PubMed

    Nejedla, Michaela; Sadi, Sara; Sulimenko, Vadym; de Almeida, Francisca Nunes; Blom, Hans; Draber, Pavel; Aspenström, Pontus; Karlsson, Roger

    2016-08-01

    Profilin controls actin nucleation and assembly processes in eukaryotic cells. Actin nucleation and elongation promoting factors (NEPFs) such as Ena/VASP, formins, and WASP-family proteins recruit profilin:actin for filament formation. Some of these are found to be microtubule associated, making actin polymerization from microtubule-associated platforms possible. Microtubules are implicated in focal adhesion turnover, cell polarity establishment, and migration, illustrating the coupling between actin and microtubule systems. Here we demonstrate that profilin is functionally linked to microtubules with formins and point to formins as major mediators of this association. To reach this conclusion, we combined different fluorescence microscopy techniques, including superresolution microscopy, with siRNA modulation of profilin expression and drug treatments to interfere with actin dynamics. Our studies show that profilin dynamically associates with microtubules and this fraction of profilin contributes to balance actin assembly during homeostatic cell growth and affects micro-tubule dynamics. Hence profilin functions as a regulator of microtubule (+)-end turnover in addition to being an actin control element. PMID:27307590

  7. Actin cytoskeleton redox proteome oxidation by cadmium

    PubMed Central

    Go, Young-Mi; Orr, Michael

    2013-01-01

    Epidemiological studies associate environmental cadmium (Cd) exposure with the risk of lung diseases. Although mechanisms are not fully elucidated, several studies demonstrate Cd effects on actin and actin-associated proteins. In a recent study of Cd at concentrations similar to environmental exposures, we found that redox-dependent inflammatory signaling by NF-κB was sensitive to the actin-disrupting agent, cytochalasin D. The goal of the present study was to use mass spectrometry-based redox proteomics to investigate Cd effects on the actin cytoskeleton proteome and related functional pathways in lung cells at low environmental concentrations. The results showed that Cd under conditions that did not alter total protein thiols or glutathione redox state caused significant oxidation of peptidyl Cys of proteins regulating actin cytoskeleton. Immunofluorescence microscopy of lung fibroblasts and pulmonary artery endothelial cells showed that low-dose Cd exposure stimulated filamentous actin formation and nuclear localization of destrin, an actin-depolymerizing factor. Taken together, the results show that redox states of peptidyl Cys in proteins associated with actin cytoskeleton pathways are selectively oxidized in lung by Cd at levels thought to occur from environmental exposure. PMID:24077948

  8. Actin motility: formin a SCAry tail.

    PubMed

    Alberts, Art; Way, Michael

    2011-01-11

    A new biochemical analysis has revealed that the Rickettsia bacterial protein Sca2--recently shown to be essential for virulence and actin-dependent motility--assembles actin filaments using a mechanism that functionally resembles the processive elongation tactics used by formins. PMID:21215933

  9. Effect of ATP on actin filament stiffness.

    PubMed

    Janmey, P A; Hvidt, S; Oster, G F; Lamb, J; Stossel, T P; Hartwig, J H

    1990-09-01

    Actin is an adenine nucleotide-binding protein and an ATPase. The bound adenine nucleotide stabilizes the protein against denaturation and the ATPase activity, although not required for actin polymerization, affects the kinetics of this assembly Here we provide evidence for another effect of adenine nucleotides. We find that actin filaments made from ATP-containing monomers, the ATPase activity of which hydrolyses ATP to ADP following polymerization, are stiff rods, whereas filaments prepared from ADP-monomers are flexible. ATP exchanges with ADP in such filaments and stiffens them. Because both kinds of actin filaments contain mainly ADP, we suggest the alignment of actin monomers in filaments that have bound and hydrolysed ATP traps them conformationally and stores elastic energy. This energy would be available for release by actin-binding proteins that transduce force or sever actin filaments. These data support earlier proposals that actin is not merely a passive cable, but has an active mechanochemical role in cell function. PMID:2168523

  10. Myelination: actin disassembly leads the way

    PubMed Central

    Samanta, Jayshree; Salzer, James L.

    2016-01-01

    The mechanisms that drive the spiral wrapping of the myelin sheath around axons are poorly understood. Two papers in this issue of Developmental Cell demonstrate that actin disassembly, rather than actin assembly, predominates during oligodendrocyte maturation and is critical for the genesis of the central myelin sheath. PMID:26218317

  11. Colchicine activates actin polymerization by microtubule depolymerization.

    PubMed

    Jung, H I; Shin, I; Park, Y M; Kang, K W; Ha, K S

    1997-06-30

    Swiss 3T3 fibroblasts were treated with the microtubule-disrupting agent colchicine to study any interaction between microtubule dynamics and actin polymerization. Colchicine increased the amount of filamentous actin (F-actin), in a dose- and time-dependent manner with a significant increase at 1 h by about 130% over control level. Confocal microscopic observation showed that colchicine increased F-actin contents by stress fiber formation without inducing membrane ruffling. Colchicine did not activate phospholipase C and phospholipase D, whereas lysophosphatidic acid did, indicating that colchicine may have a different mechanism of actin polymerization regulation from LPA. A variety of microtubule-disrupting agents stimulated actin polymerization in Swiss 3T3 and Rat-2 fibroblasts as did colchicine, but the microtubule-stabilizing agent taxol inhibited actin polymerization induced by the above microtubule-disrupting agents. In addition, colchicine-induced actin polymerization was blocked by two protein phosphatase inhibitors, okadaic acid and calyculin A. These results suggest that microtubule depolymerization activates stress fiber formation by serine/threonine dephosphorylation in fibroblasts. PMID:9264034

  12. Docking, molecular dynamics and QM/MM studies to delineate the mode of binding of CucurbitacinE to F-actin.

    PubMed

    Kumar, R Pravin; Roopa, L; Nongthomba, Upendra; Sudheer Mohammed, M M; Kulkarni, Naveen

    2016-01-01

    CucurbitacinE (CurE) has been known to bind covalently to F-actin and inhibit depolymerization. However, the mode of binding of CurE to F-actin and the consequent changes in the F-actin dynamics have not been studied. Through quantum mechanical/molecular mechanical (QM/MM) and density function theory (DFT) simulations after the molecular dynamics (MD) simulations of the docked complex of F-actin and CurE, a detailed transition state (TS) model for the Michael reaction is proposed. The TS model shows nucleophilic attack of the sulphur of Cys257 at the β-carbon of Michael Acceptor of CurE producing an enol intermediate that forms a covalent bond with CurE. The MD results show a clear difference between the structure of the F-actin in free form and F-actin complexed with CurE. CurE affects the conformation of the nucleotide binding pocket increasing the binding affinity between F-actin and ADP, which in turn could affect the nucleotide exchange. CurE binding also limits the correlated displacement of the relatively flexible domain 1 of F-actin causing the protein to retain a flat structure and to transform into a stable "tense" state. This structural transition could inhibit depolymerization of F-actin. In conclusion, CurE allosterically modulates ADP and stabilizes F-actin structure, thereby affecting nucleotide exchange and depolymerization of F-actin. PMID:26615469

  13. Actin-binding proteins: the long road to understanding the dynamic landscape of cellular actin networks.

    PubMed

    Lappalainen, Pekka

    2016-08-15

    The actin cytoskeleton supports a vast number of cellular processes in nonmuscle cells. It is well established that the organization and dynamics of the actin cytoskeleton are controlled by a large array of actin-binding proteins. However, it was only 40 years ago that the first nonmuscle actin-binding protein, filamin, was identified and characterized. Filamin was shown to bind and cross-link actin filaments into higher-order structures and contribute to phagocytosis in macrophages. Subsequently many other nonmuscle actin-binding proteins were identified and characterized. These proteins regulate almost all steps of the actin filament assembly and disassembly cycles, as well as the arrangement of actin filaments into diverse three-dimensional structures. Although the individual biochemical activities of most actin-regulatory proteins are relatively well understood, knowledge of how these proteins function together in a common cytoplasm to control actin dynamics and architecture is only beginning to emerge. Furthermore, understanding how signaling pathways and mechanical cues control the activities of various actin-binding proteins in different cellular, developmental, and pathological processes will keep researchers busy for decades. PMID:27528696

  14. Xenopus egg cytoplasm with intact actin.

    PubMed

    Field, Christine M; Nguyen, Phuong A; Ishihara, Keisuke; Groen, Aaron C; Mitchison, Timothy J

    2014-01-01

    We report optimized methods for preparing Xenopus egg extracts without cytochalasin D, that we term "actin-intact egg extract." These are undiluted egg cytoplasm that contains abundant organelles, and glycogen which supplies energy, and represents the least perturbed cell-free cytoplasm preparation we know of. We used this system to probe cell cycle regulation of actin and myosin-II dynamics (Field et al., 2011), and to reconstitute the large, interphase asters that organize early Xenopus embryos (Mitchison et al., 2012; Wühr, Tan, Parker, Detrich, & Mitchison, 2010). Actin-intact Xenopus egg extracts are useful for analysis of actin dynamics, and interaction of actin with other cytoplasmic systems, in a cell-free system that closely mimics egg physiology, and more generally for probing the biochemistry and biophysics of the egg, zygote, and early embryo. Detailed protocols are provided along with assays used to check cell cycle state and tips for handling and storing undiluted egg extracts. PMID:24630119

  15. Quantification of Canopy Structure and its Implication on Radiative Transfer, Carbon Dioxide and Energy Flux Densities in a Heterogeneous Oak-Grass Savanna Ecosystem at the Landscape Scale

    NASA Astrophysics Data System (ADS)

    Sonnentag, O.; Ryu, Y.; Vargas, R.; Baldocchi, D.

    2008-12-01

    Oak-grass savanna ecosystems are characterized by pronounced heterogeneity in canopy structure at the landscape scale. Due to this heterogeneity the accurate quantification of canopy structure still remains a major challenge. The objectives of this study are to quantify clumping index, leaf area index (LAI) and the leaf inclination angle distribution function (LIADF) to describe the canopy structure of an oak-grass savanna ecosystem in California, USA. This information is critical for utilizing a radiative transfer model to compute CO2 and energy flux densities. We used four established techniques (LAI-2000 Plant Canopy Analyzer, digital hemispherical photography, the Tracing Radiation and Architecture of Canopies (TRAC) instrument, and a robotics railroad radiometer) to measure clumping index and LAI within a 300 x 300 m plot centered at an eddy covariance (EC) tower. Leaf inclination angle distributions were assessed from digital photographs at multiple height intervals through analysis with a public domain image processing software. Preliminary analysis of the data showed that mean values for clumping index and LAI obtained from the various instruments are in good agreement, thus reducing the uncertainty inherent in the measurements. Our leaf angle measurements revealed the canopy to be predominantly erectophile at all height intervals, an ecological consequence of the fact that oak leaves must be erect to reduce thermal load.

  16. Effects of magnetic flux density and substrate bias voltage on Ni films prepared on a flexible substrate material using unbalanced magnetron sputtering assisted by inductively coupled plasma

    SciTech Connect

    Koda, Tatsunori; Toyota, Hiroshi

    2014-03-15

    The authors fabricated Ni films on a flexible substrate material using unbalanced magnetron sputtering assisted by inductively coupled plasma. The effects of magnetic flux density B{sub C} and substrate DC bias voltage V{sub S} on the Ni film structures were investigated. For V{sub S} = −40 V, the average surface grain size D{sub G} measured by atomic force microscopy for B{sub C} = 0, 3, and 5 mT was 88.2, 95.4, and 104.4 nm, respectively. In addition, D{sub G} increased with V{sub S}. From x-ray diffraction measurements, the (111) and (200) peaks were clearly visible for the fabricated Ni films. The ratio of the integrated intensities of I(111)/I(200) increased with V{sub S}. For V{sub S} = −40 V and B{sub C} = 3 mT, a film resistivity ρ of 8.96 × 10{sup −6} Ω cm was observed, which is close to the Ni bulk value of 6.84 × 10{sup −6} Ω cm. From these results, the authors determined that the structure of the fabricated Ni films on the flexible substrate material was affected by the values of B{sub C} and V{sub S}.

  17. Determination of neutron flux density distribution in the core with LEU fuel IRT-4M at the training reactor VR-1

    SciTech Connect

    Huml, O.

    2008-07-15

    The objective of this work was to determine the neutron flux density distribution in various places of the training reactor VR-1 Sparrow. This experiment was performed on the new core design C1, composed of the new low-enriched uranium fuel cells IRT-4M (19.7 %). This fuel replaced the old high-enriched uranium fuel IRT-3M (36 %) within the framework of the RERTR Program in September 2005. The measurement used the neutron activation analysis method with gold wires. The principle of this method consists in neutron capture in a nucleus of the material forming the activation detector. This capture can change the nucleus in a radioisotope, whose activity can be measured. The absorption cross-section values were evaluated by MCNP computer code. The gold wires were irradiated in seven different positions in the core C1. All irradiations were performed at reactor power level 1E8 (1 kW{sub therm}). The activity of segments of irradiated wires was measured by special automatic device called 'Drat' (Wire in English). (author)

  18. Actin dynamics shape microglia effector functions.

    PubMed

    Uhlemann, Ria; Gertz, Karen; Boehmerle, Wolfgang; Schwarz, Tobias; Nolte, Christiane; Freyer, Dorette; Kettenmann, Helmut; Endres, Matthias; Kronenberg, Golo

    2016-06-01

    Impaired actin filament dynamics have been associated with cellular senescence. Microglia, the resident immune cells of the brain, are emerging as a central pathophysiological player in neurodegeneration. Microglia activation, which ranges on a continuum between classical and alternative, may be of critical importance to brain disease. Using genetic and pharmacological manipulations, we studied the effects of alterations in actin dynamics on microglia effector functions. Disruption of actin dynamics did not affect transcription of genes involved in the LPS-triggered classical inflammatory response. By contrast, in consequence of impaired nuclear translocation of phospho-STAT6, genes involved in IL-4 induced alternative activation were strongly downregulated. Functionally, impaired actin dynamics resulted in reduced NO secretion and reduced release of TNFalpha and IL-6 from LPS-stimulated microglia and of IGF-1 from IL-4 stimulated microglia. However, pathological stabilization of the actin cytoskeleton increased LPS-induced release of IL-1beta and IL-18, which belong to an unconventional secretory pathway. Reduced NO release was associated with decreased cytoplasmic iNOS protein expression and decreased intracellular arginine uptake. Furthermore, disruption of actin dynamics resulted in reduced microglia migration, proliferation and phagocytosis. Finally, baseline and ATP-induced [Ca(2+)]int levels were significantly increased in microglia lacking gelsolin, a key actin-severing protein. Together, the dynamic state of the actin cytoskeleton profoundly and distinctly affects microglia behaviours. Disruption of actin dynamics attenuates M2 polarization by inhibiting transcription of alternative activation genes. In classical activation, the role of actin remodelling is complex, does not relate to gene transcription and shows a major divergence between cytokines following conventional and unconventional secretion. PMID:25989853

  19. Probing actin incorporation into myofibrils using Asp11 and His73 actin mutants.

    PubMed

    Xia, D; Peng, B; Sesok, D A; Peng, I

    1993-01-01

    We used a cell free system Bouché et al.: J. Cell Biol. 107:587-596, 1988] to study the incorporation of actin into myofibrils. We used alpha-skeletal muscle actin and actins with substitutions of either His73 [Solomon and Rubenstein: J. Biol.Chem. 262:11382, 1987], or Asp11 [Solomon et al.: J. Biol. Chem. 263:19662, 1988]. Actins were translated in reticulocyte lysate and incubated with myofibrils. The incorporated wild type actin could be cross-linked into dimers using N,N'-1,4-phenylenebismaleimide (PBM), indicating that the incorporated actin is actually inserted into the thin filaments of the myofibril. The His73 mutants incorporated to the same extent as wild type actin and was also cross-linked with PBM. Although some of the Asp11 mutants co-assembled with carrier actin, only 1-3% of the Asp11 mutant actins incorporated after 2 min and did not increase after 2 hr. Roughly 17% of wild type actin incorporated after 2 min and 31% after 2 hr. ATP increased the release of wild type actin from myofibrils, but did not increase the release of Asp11 mutants. We suggest that (1) the incorporation of wild type and His73 mutant actins was due to a physiological process whereas association of Asp11 mutants with myofibrils was non-specific, (2) the incorporation of wild type actin involved a rapid initial phase, followed by a slower phase, and (3) since some of the Asp11 mutants can co-assemble with wild type actin, the ability to self-assemble was not sufficient for incorporation into myofibrils. Thus, incorporation probably includes interaction between actin and a thin filament associated protein. We also showed that incorporation occurred at actin concentrations which would cause disassembly of F-actin. Since the myofibrils did not show large scale disassembly but incorporated actin, filament stability and monomer incorporation are likely to be mediated by actin associated proteins of the myofibril. PMID:8287497

  20. Dynamics of an actin spring

    NASA Astrophysics Data System (ADS)

    Riera, Christophe; Mahadevan, L.; Shin, Jennifer; Matsudaira, Paul

    2003-03-01

    The acrosome of the sperm of the horseshoe crab (Limulus Polyphemus) is an unusual actin based system that shows a spectacular dynamical transition in the presence of Ca++ that is present in abundance in the neighborhood of the egg. During this process, the bundle, which is initially bent and twisted uncoils and becomes straight in a matter of a few seconds. Based on microstructural data, we propose a model for the dynamics of uncoiling that is best represented by a triple-well potential corresponding to the different structural arrangements of the supertwisted filaments. Each of the false, true and coiled states corresponds to a local minimum of the energy, with the true state being the one with the lowest energy. Using an evolution equation derived by balancing torques, we investigate the nucleation and propagation of the phase transition and compare the results with those of experiments. Our model quantifies the hypothesis that the acrosomal bundle behaves like a mechano-chemical spring.

  1. Binding of WIP to Actin Is Essential for T Cell Actin Cytoskeleton Integrity and Tissue Homing

    PubMed Central

    Massaad, Michel J.; Oyoshi, Michiko K.; Kane, Jennifer; Koduru, Suresh; Alcaide, Pilar; Nakamura, Fumihiko; Ramesh, Narayanaswamy; Luscinskas, Francis W.; Hartwig, John

    2014-01-01

    The Wiskott-Aldrich syndrome protein (WASp) is important for actin polymerization in T cells and for their migration. WASp-interacting protein (WIP) binds to and stabilizes WASp and also interacts with actin. Cytoskeletal and functional defects are more severe in WIP−/− T cells, which lack WASp, than in WASp−/− T cells, suggesting that WIP interaction with actin may be important for T cell cytoskeletal integrity and function. We constructed mice that lack the actin-binding domain of WIP (WIPΔABD mice). WIPΔABD associated normally with WASp but not F-actin. T cells from WIPΔABD mice had normal WASp levels but decreased cellular F-actin content, a disorganized actin cytoskeleton, impaired chemotaxis, and defective homing to lymph nodes. WIPΔABD mice exhibited a T cell intrinsic defect in contact hypersensitivity and impaired responses to cutaneous challenge with protein antigen. Adoptively transferred antigen-specific CD4+ T cells from WIPΔABD mice had decreased homing to antigen-challenged skin of wild-type recipients. These findings show that WIP binding to actin, independently of its binding to WASp, is critical for the integrity of the actin cytoskeleton in T cells and for their migration into tissues. Disruption of WIP binding to actin could be of therapeutic value in T cell-driven inflammatory diseases. PMID:25246631

  2. Actin-curcumin interaction: insights into the mechanism of actin polymerization inhibition.

    PubMed

    Dhar, Gopa; Chakravarty, Devlina; Hazra, Joyita; Dhar, Jesmita; Poddar, Asim; Pal, Mahadeb; Chakrabarti, Pinak; Surolia, Avadhesha; Bhattacharyya, Bhabatarak

    2015-02-01

    Curcumin, derived from rhizomes of the Curcuma longa plant, is known to possess a wide range of medicinal properties. We have examined the interaction of curcumin with actin and determined their binding and thermodynamic parameters using isothermal titration calorimetry. Curcumin is weakly fluorescent in aqueous solution, and binding to actin enhances fluorescence several fold with a large blue shift in the emission maximum. Curcumin inhibits microfilament formation, which is similar to its role in inhibiting microtubule formation. We synthesized a series of stable curcumin analogues to examine their affinity for actin and their ability to inhibit actin self-assembly. Results show that curcumin is a ligand with two symmetrical halves, each of which possesses no activity individually. Oxazole, pyrazole, and acetyl derivatives are less effective than curcumin at inhibiting actin self-assembly, whereas a benzylidiene derivative is more effective. Cell biology studies suggest that disorganization of the actin network leads to destabilization of filaments in the presence of curcumin. Molecular docking reveals that curcumin binds close to the cytochalasin binding site of actin. Further molecular dynamics studies reveal a possible allosteric effect in which curcumin binding at the "barbed end" of actin is transmitted to the "pointed end", where conformational changes disrupt interactions with the adjacent actin monomer to interrupt filament formation. Finally, the recognition and binding of actin by curcumin is yet another example of its unique ability to target multiple receptors. PMID:25564154

  3. Nuclear actin and myosins in adenovirus infection.

    PubMed

    Fuchsova, Beata; Serebryannyy, Leonid A; de Lanerolle, Primal

    2015-11-01

    Adenovirus serotypes have been shown to cause drastic changes in nuclear organization, including the transcription machinery, during infection. This ability of adenovirus to subvert transcription in the host cell facilitates viral replication. Because nuclear actin and nuclear myosin I, myosin V and myosin VI have been implicated as direct regulators of transcription and important factors in the replication of other viruses, we sought to determine how nuclear actin and myosins are involved in adenovirus infection. We first confirmed reorganization of the host's transcription machinery to viral replication centers. We found that nuclear actin also reorganizes to sites of transcription through the intermediate but not the advanced late phase of viral infection. Furthermore, nuclear myosin I localized with nuclear actin and sites of transcription in viral replication centers. Intriguingly, nuclear myosins V and VI, which also reorganized to viral replication centers, exhibited different localization patterns, suggesting specialized roles for these nuclear myosins. Finally, we assessed the role of actin in adenovirus infection and found both cytoplasmic and nuclear actin likely play roles in adenovirus infection and replication. Together our data suggest the involvement of actin and multiple myosins in the nuclear replication and late viral gene expression of adenovirus. PMID:26226218

  4. Erbium laser resurfacing for actinic cheilitis.

    PubMed

    Cohen, Joel L

    2013-11-01

    Actinic cheilitis is a precancerous condition characterized by grayish-whitish area(s) of discoloration on the mucosal lip, often blunting the demarcation between mucosa and cutaneous lip. Actinic cheilitis is considered to be an early part of the spectrum of squamous cell carcinoma. Squamous cell carcinoma specifically of the lip has a high rate of recurrence and metastasis through the oral cavity leading to a poor overall survival. Risk factors for the development of actinic cheilitis include chronic solar irradiation, increasing age, male gender, light skin complexion, immunosuppression, and possibly tobacco and alcohol consumption. Treatment options include topical pharmacotherapy (eg, fluorouracil, imiquimod) or procedural interventions (eg, cryotherapy, electrosurgery, surgical vermillionectomy, laser resurfacing), each with their known advantages and disadvantages. There is little consensus as to which treatment options offer the most clinical utility given the paucity of comparative clinical data. In my practice, laser resurfacing has become an important tool for the treatment of actinic cheilitis owing to its ease of use and overall safety, tolerability, and cosmetic acceptability. Herein the use of erbium laser resurfacing is described for three actinic cheilitis presentations for which I find it particularly useful: clinically prominent actinic cheilitis, biopsy-proven actinic cheilitis, and treatment of the entire lip following complete tumor excision of squamous cell carcinoma. All patients were treated with a 2940-nm erbium laser (Sciton Profile Contour Tunable Resurfacing Laser [TRL], Sciton, Inc., Palo Alto, CA). PMID:24196339

  5. Antibodies covalently immobilized on actin filaments for fast myosin driven analyte transport.

    PubMed

    Kumar, Saroj; ten Siethoff, Lasse; Persson, Malin; Lard, Mercy; te Kronnie, Geertruy; Linke, Heiner; Månsson, Alf

    2012-01-01

    Biosensors would benefit from further miniaturization, increased detection rate and independence from external pumps and other bulky equipment. Whereas transportation systems built around molecular motors and cytoskeletal filaments hold significant promise in the latter regard, recent proof-of-principle devices based on the microtubule-kinesin motor system have not matched the speed of existing methods. An attractive solution to overcome this limitation would be the use of myosin driven propulsion of actin filaments which offers motility one order of magnitude faster than the kinesin-microtubule system. Here, we realized a necessary requirement for the use of the actomyosin system in biosensing devices, namely covalent attachment of antibodies to actin filaments using heterobifunctional cross-linkers. We also demonstrated consistent and rapid myosin II driven transport where velocity and the fraction of motile actin filaments was negligibly affected by the presence of antibody-antigen complexes at rather high density (>20 µm(-1)). The results, however, also demonstrated that it was challenging to consistently achieve high density of functional antibodies along the actin filament, and optimization of the covalent coupling procedure to increase labeling density should be a major focus for future work. Despite the remaining challenges, the reported advances are important steps towards considerably faster nanoseparation than shown for previous molecular motor based devices, and enhanced miniaturization because of high bending flexibility of actin filaments. PMID:23056279

  6. Antibodies Covalently Immobilized on Actin Filaments for Fast Myosin Driven Analyte Transport

    PubMed Central

    Kumar, Saroj; ten Siethoff, Lasse; Persson, Malin; Lard, Mercy; te Kronnie, Geertruy; Linke, Heiner; Månsson, Alf

    2012-01-01

    Biosensors would benefit from further miniaturization, increased detection rate and independence from external pumps and other bulky equipment. Whereas transportation systems built around molecular motors and cytoskeletal filaments hold significant promise in the latter regard, recent proof-of-principle devices based on the microtubule-kinesin motor system have not matched the speed of existing methods. An attractive solution to overcome this limitation would be the use of myosin driven propulsion of actin filaments which offers motility one order of magnitude faster than the kinesin-microtubule system. Here, we realized a necessary requirement for the use of the actomyosin system in biosensing devices, namely covalent attachment of antibodies to actin filaments using heterobifunctional cross-linkers. We also demonstrated consistent and rapid myosin II driven transport where velocity and the fraction of motile actin filaments was negligibly affected by the presence of antibody-antigen complexes at rather high density (>20 µm−1). The results, however, also demonstrated that it was challenging to consistently achieve high density of functional antibodies along the actin filament, and optimization of the covalent coupling procedure to increase labeling density should be a major focus for future work. Despite the remaining challenges, the reported advances are important steps towards considerably faster nanoseparation than shown for previous molecular motor based devices, and enhanced miniaturization because of high bending flexibility of actin filaments. PMID:23056279

  7. Actinic Granuloma with Focal Segmental Glomerulosclerosis

    PubMed Central

    Phasukthaworn, Ruedee; Chanprapaph, Kumutnart; Vachiramon, Vasanop

    2016-01-01

    Actinic granuloma is an uncommon granulomatous disease, characterized by annular erythematous plaque with central clearing predominately located on sun-damaged skin. The pathogenesis is not well understood, ultraviolet radiation is recognized as precipitating factor. We report a case of a 52-year-old woman who presented with asymptomatic annular erythematous plaques on the forehead and both cheeks persisting for 2 years. The clinical presentation and histopathologic findings support the diagnosis of actinic granuloma. During that period of time, she also developed focal segmental glomerulosclerosis. The association between actinic granuloma and focal segmental glomerulosclerosis needs to be clarified by further studies. PMID:27293392

  8. Dynamic reorganization of the actin cytoskeleton

    PubMed Central

    Gressin, Laurène; Théry, Manuel; Blanchoin, Laurent

    2015-01-01

    Cellular processes, including morphogenesis, polarization, and motility, rely on a variety of actin-based structures. Although the biochemical composition and filament organization of these structures are different, they often emerge from a common origin. This is possible because the actin structures are highly dynamic. Indeed, they assemble, grow, and disassemble in a time scale of a second to a minute. Therefore, the reorganization of a given actin structure can promote the formation of another. Here, we discuss such transitions and illustrate them with computer simulations. PMID:26989473

  9. Binding of actin to lens alpha crystallins

    NASA Technical Reports Server (NTRS)

    Gopalakrishnan, S.; Takemoto, L.; Spooner, B. S. (Principal Investigator)

    1992-01-01

    Actin has been coupled to a cyanogen bromide-activated Sepharose 4B column, then tested for binding to alpha, beta, and gamma crystallin preparations from the bovine lens. Alpha, but not beta or gamma, crystallins bound to the actin affinity column in a time dependent and saturable manner. Subfractionation of the alpha crystallin preparation into the alpha-A and alpha-B species, followed by incubation with the affinity column, demonstrated that both species bound approximately the same. Together, these studies demonstrate a specific and saturable binding of lens alpha-A and alpha-B with actin.

  10. Adhesion controls bacterial actin polymerization-based movement.

    PubMed

    Soo, Frederick S; Theriot, Julie A

    2005-11-01

    As part of its infectious life cycle, the bacterial pathogen Listeria monocytogenes propels itself through the host-cell cytoplasm by triggering the polymerization of host-cell actin near the bacterial surface, harnessing the activity of several cytoskeletal proteins used during actin-based cell crawling. To distinguish among several classes of biophysical models of actin-based bacterial movement, we used a high-throughput tracking technique to record the movement of many individual bacteria during temperature shifts. The speed of each bacterium varied strongly with temperature, closely following the Arrhenius rate law. Among bacteria, the prefactor A of the Arrhenius dependence unexpectedly varied exponentially with apparent activation energy, E(a), over a wide range (8-21 kcal/mol), reminiscent of the "rate compensation effect" of classical catalytic reactions. Average E(a) were increased for mutant bacteria deficient in binding Ena/VASP proteins and bacteria moving in diluted extract. These two effects were additive. The observed temperature and rate compensation effects are consistent with a class of simple kinetic models in which the bacterium advances through the thermally driven, cooperative breakage of groups of adhesive bonds on its surface. The estimated number of coupled adhesive bonds N on the bacterial surface varies between 10 and 40 bonds. In contrast to other models, this model correctly predicts an experimentally observed negative correlation between bacterial speed and actin gel density. The idea that speed depends on adhesion, rather than polymerization, suggests several alternative mechanisms by which known cytoskeletal regulatory proteins could control cellular movement. PMID:16251274

  11. Actin Depletion Initiates Events Leading to Granule Secretion at the Immunological Synapse

    PubMed Central

    Ritter, Alex T.; Asano, Yukako; Stinchcombe, Jane C.; Dieckmann, N.M.G.; Chen, Bi-Chang; Gawden-Bone, C.; van Engelenburg, Schuyler; Legant, Wesley; Gao, Liang; Davidson, Michael W.; Betzig, Eric; Lippincott-Schwartz, Jennifer; Griffiths, Gillian M.

    2015-01-01

    Summary Cytotoxic T lymphocytes (CTLs) use polarized secretion to rapidly destroy virally infected and tumor cells. To understand the temporal relationships between key events leading to secretion, we used high-resolution 4D imaging. CTLs approached targets with actin-rich projections at the leading edge, creating an initially actin-enriched contact with rearward-flowing actin. Within 1 min, cortical actin reduced across the synapse, T cell receptors (TCRs) clustered centrally to form the central supramolecular activation cluster (cSMAC), and centrosome polarization began. Granules clustered around the moving centrosome within 2.5 min and reached the synapse after 6 min. TCR-bearing intracellular vesicles were delivered to the cSMAC as the centrosome docked. We found that the centrosome and granules were delivered to an area of membrane with reduced cortical actin density and phospholipid PIP2. These data resolve the temporal order of events during synapse maturation in 4D and reveal a critical role for actin depletion in regulating secretion. PMID:25992860

  12. Molecular Architecture of Synaptic Actin Cytoskeleton in Hippocampal Neurons Reveals a Mechanism of Dendritic Spine Morphogenesis

    PubMed Central

    Korobova, Farida

    2010-01-01

    Excitatory synapses in the brain play key roles in learning and memory. The formation and functions of postsynaptic mushroom-shaped structures, dendritic spines, and possibly of presynaptic terminals, rely on actin cytoskeleton remodeling. However, the cytoskeletal architecture of synapses remains unknown hindering the understanding of synapse morphogenesis. Using platinum replica electron microscopy, we characterized the cytoskeletal organization and molecular composition of dendritic spines, their precursors, dendritic filopodia, and presynaptic boutons. A branched actin filament network containing Arp2/3 complex and capping protein was a dominant feature of spine heads and presynaptic boutons. Surprisingly, the spine necks and bases, as well as dendritic filopodia, also contained a network, rather than a bundle, of branched and linear actin filaments that was immunopositive for Arp2/3 complex, capping protein, and myosin II, but not fascin. Thus, a tight actin filament bundle is not necessary for structural support of elongated filopodia-like protrusions. Dynamically, dendritic filopodia emerged from densities in the dendritic shaft, which by electron microscopy contained branched actin network associated with dendritic microtubules. We propose that dendritic spine morphogenesis begins from an actin patch elongating into a dendritic filopodium, which tip subsequently expands via Arp2/3 complex-dependent nucleation and which length is modulated by myosin II-dependent contractility. PMID:19889835

  13. Computational model of polarized actin cables and cytokinetic actin ring formation in budding yeast

    PubMed Central

    Tang, Haosu; Bidone, Tamara C.

    2015-01-01

    The budding yeast actin cables and contractile ring are important for polarized growth and division, revealing basic aspects of cytoskeletal function. To study these formin-nucleated structures, we built a 3D computational model with actin filaments represented as beads connected by springs. Polymerization by formins at the bud tip and bud neck, crosslinking, severing, and myosin pulling, are included. Parameter values were estimated from prior experiments. The model generates actin cable structures and dynamics similar to those of wild type and formin deletion mutant cells. Simulations with increased polymerization rate result in long, wavy cables. Simulated pulling by type V myosin stretches actin cables. Increasing the affinity of actin filaments for the bud neck together with reduced myosin V pulling promotes the formation of a bundle of antiparallel filaments at the bud neck, which we suggest as a model for the assembly of actin filaments to the contractile ring. PMID:26538307

  14. Electron Tomography of Cryofixed, Isometrically Contracting Insect Flight Muscle Reveals Novel Actin-Myosin Interactions

    PubMed Central

    Wu, Shenping; Liu, Jun; Tregear, Richard T.; Winkler, Hanspeter; Franzini-Armstrong, Clara; Sasaki, Hiroyuki; Lucaveche, Carmen; Goldman, Yale E.; Reedy, Michael K.; Taylor, Kenneth A.

    2010-01-01

    Background Isometric muscle contraction, where force is generated without muscle shortening, is a molecular traffic jam in which the number of actin-attached motors is maximized and all states of motor action are trapped with consequently high heterogeneity. This heterogeneity is a major limitation to deciphering myosin conformational changes in situ. Methodology We used multivariate data analysis to group repeat segments in electron tomograms of isometrically contracting insect flight muscle, mechanically monitored, rapidly frozen, freeze substituted, and thin sectioned. Improved resolution reveals the helical arrangement of F-actin subunits in the thin filament enabling an atomic model to be built into the thin filament density independent of the myosin. Actin-myosin attachments can now be assigned as weak or strong by their motor domain orientation relative to actin. Myosin attachments were quantified everywhere along the thin filament including troponin. Strong binding myosin attachments are found on only four F-actin subunits, the “target zone”, situated exactly midway between successive troponin complexes. They show an axial lever arm range of 77°/12.9 nm. The lever arm azimuthal range of strong binding attachments has a highly skewed, 127° range compared with X-ray crystallographic structures. Two types of weak actin attachments are described. One type, found exclusively in the target zone, appears to represent pre-working-stroke intermediates. The other, which contacts tropomyosin rather than actin, is positioned M-ward of the target zone, i.e. the position toward which thin filaments slide during shortening. Conclusion We present a model for the weak to strong transition in the myosin ATPase cycle that incorporates azimuthal movements of the motor domain on actin. Stress/strain in the S2 domain may explain azimuthal lever arm changes in the strong binding attachments. The results support previous conclusions that the weak attachments preceding force

  15. The Plant Actin Cytoskeleton Responds to Signals from Microbe-Associated Molecular Patterns

    PubMed Central

    Henty-Ridilla, Jessica L.; Shimono, Masaki; Li, Jiejie; Chang, Jeff H.; Day, Brad; Staiger, Christopher J.

    2013-01-01

    Plants are constantly exposed to a large and diverse array of microbes; however, most plants are immune to the majority of potential invaders and susceptible to only a small subset of pathogens. The cytoskeleton comprises a dynamic intracellular framework that responds rapidly to biotic stresses and supports numerous fundamental cellular processes including vesicle trafficking, endocytosis and the spatial distribution of organelles and protein complexes. For years, the actin cytoskeleton has been assumed to play a role in plant innate immunity against fungi and oomycetes, based largely on static images and pharmacological studies. To date, however, there is little evidence that the host-cell actin cytoskeleton participates in responses to phytopathogenic bacteria. Here, we quantified the spatiotemporal changes in host-cell cytoskeletal architecture during the immune response to pathogenic and non-pathogenic strains of Pseudomonas syringae pv. tomato DC3000. Two distinct changes to host cytoskeletal arrays were observed that correspond to distinct phases of plant-bacterial interactions i.e. the perception of microbe-associated molecular patterns (MAMPs) during pattern-triggered immunity (PTI) and perturbations by effector proteins during effector-triggered susceptibility (ETS). We demonstrate that an immediate increase in actin filament abundance is a conserved and novel component of PTI. Notably, treatment of leaves with a MAMP peptide mimic was sufficient to elicit a rapid change in actin organization in epidermal cells, and this actin response required the host-cell MAMP receptor kinase complex, including FLS2, BAK1 and BIK1. Finally, we found that actin polymerization is necessary for the increase in actin filament density and that blocking this increase with the actin-disrupting drug latrunculin B leads to enhanced susceptibility of host plants to pathogenic and non-pathogenic bacteria. PMID:23593000

  16. Electron Tomography of Cryofixed, Isometrically Contracting Insect Flight Muscle Reveals Novel Actin-Myosin Interactions

    SciTech Connect

    Wu, Shenping; Liu, Jun; Reedy, Mary C.; Tregear, Richard T.; Winkler, Hanspeter; Franzini-Armstrong, Clara; Sasaki, Hiroyuki; Lucaveche, Carmen; Goldman, Yale E.; Reedy, Michael K.; Taylor, Kenneth A.

    2010-10-22

    Isometric muscle contraction, where force is generated without muscle shortening, is a molecular traffic jam in which the number of actin-attached motors is maximized and all states of motor action are trapped with consequently high heterogeneity. This heterogeneity is a major limitation to deciphering myosin conformational changes in situ. We used multivariate data analysis to group repeat segments in electron tomograms of isometrically contracting insect flight muscle, mechanically monitored, rapidly frozen, freeze substituted, and thin sectioned. Improved resolution reveals the helical arrangement of F-actin subunits in the thin filament enabling an atomic model to be built into the thin filament density independent of the myosin. Actin-myosin attachments can now be assigned as weak or strong by their motor domain orientation relative to actin. Myosin attachments were quantified everywhere along the thin filament including troponin. Strong binding myosin attachments are found on only four F-actin subunits, the 'target zone', situated exactly midway between successive troponin complexes. They show an axial lever arm range of 77{sup o}/12.9 nm. The lever arm azimuthal range of strong binding attachments has a highly skewed, 127{sup o} range compared with X-ray crystallographic structures. Two types of weak actin attachments are described. One type, found exclusively in the target zone, appears to represent pre-working-stroke intermediates. The other, which contacts tropomyosin rather than actin, is positioned M-ward of the target zone, i.e. the position toward which thin filaments slide during shortening. We present a model for the weak to strong transition in the myosin ATPase cycle that incorporates azimuthal movements of the motor domain on actin. Stress/strain in the S2 domain may explain azimuthal lever arm changes in the strong binding attachments. The results support previous conclusions that the weak attachments preceding force generation are very

  17. The Tyrosine Kinase Activity of c-Src Regulates Actin Dynamics and Organization of Podosomes in Osteoclasts

    PubMed Central

    Destaing, Olivier; Sanjay, Archana; Itzstein, Cecile; Horne, William C.; Toomre, Derek

    2008-01-01

    Podosomes are dynamic actin-rich structures composed of a dense F-actin core surrounded by a cloud of more diffuse F-actin. Src performs one or more unique functions in osteoclasts (OCLs), and podosome belts and bone resorption are impaired in the absence of Src. Using Src−/− OCLs, we investigated the specific functions of Src in the organization and dynamics of podosomes. We found that podosome number and the podosome-associated actin cloud were decreased in Src−/− OCLs. Videomicroscopy and fluorescence recovery after photobleaching analysis revealed that the life span of Src−/− podosomes was increased fourfold and that the rate of actin flux in the core was decreased by 40%. Thus, Src regulates the formation, structure, life span, and rate of actin polymerization in podosomes and in the actin cloud. Rescue of Src−/− OCLs with Src mutants showed that both the kinase activity and either the SH2 or the SH3 binding domain are required for Src to restore normal podosome organization and dynamics. Moreover, inhibition of Src family kinase activities in Src−/− OCLs by Src inhibitors or by expressing dominant-negative SrcK295M induced the formation of abnormal podosomes. Thus, Src is an essential regulator of podosome structure, dynamics and organization. PMID:17978100

  18. Regulation of actin polymerization by tropomodulin-3 controls megakaryocyte actin organization and platelet biogenesis.

    PubMed

    Sui, Zhenhua; Nowak, Roberta B; Sanada, Chad; Halene, Stephanie; Krause, Diane S; Fowler, Velia M

    2015-07-23

    The actin cytoskeleton is important for platelet biogenesis. Tropomodulin-3 (Tmod3), the only Tmod isoform detected in platelets and megakaryocytes (MKs), caps actin filament (F-actin) pointed ends and binds tropomyosins (TMs), regulating actin polymerization and stability. To determine the function of Tmod3 in platelet biogenesis, we studied Tmod3(-/-) embryos, which are embryonic lethal by E18.5. Tmod3(-/-) embryos often show hemorrhaging at E14.5 with fewer and larger platelets, indicating impaired platelet biogenesis. MK numbers are moderately increased in Tmod3(-/-) fetal livers, with only a slight increase in the 8N population, suggesting that MK differentiation is not significantly affected. However, Tmod3(-/-) MKs fail to develop a normal demarcation membrane system (DMS), and cytoplasmic organelle distribution is abnormal. Moreover, cultured Tmod3(-/-) MKs exhibit impaired proplatelet formation with a wide range of proplatelet bud sizes, including abnormally large proplatelet buds containing incorrect numbers of von Willebrand factor-positive granules. Tmod3(-/-) MKs exhibit F-actin disturbances, and Tmod3(-/-) MKs spreading on collagen fail to polymerize F-actin into actomyosin contractile bundles. Tmod3 associates with TM4 and the F-actin cytoskeleton in wild-type MKs, and confocal microscopy reveals that Tmod3, TM4, and F-actin partially colocalize near the membrane of proplatelet buds. In contrast, the abnormally large proplatelets from Tmod3(-/-) MKs show increased F-actin and redistribution of F-actin and TM4 from the cortex to the cytoplasm, but normal microtubule coil organization. We conclude that F-actin capping by Tmod3 regulates F-actin organization in mouse fetal liver-derived MKs, thereby controlling MK cytoplasmic morphogenesis, including DMS formation and organelle distribution, as well as proplatelet formation and sizing. PMID:25964668

  19. Nuclear actin levels as an important transcriptional switch

    PubMed Central

    Huet, Guillaume; Skarp, Kari-Pekka; Vartiainen, Maria K.

    2012-01-01

    Nuclear actin levels have recently been linked to different cellular fates, suggesting that actin could act as a switch between altered transcriptional states. Here we discuss our latest results on the mechanisms by which nuclear actin levels are regulated and their implications to the functional significance of nuclear actin. PMID:22771994

  20. Genetics Home Reference: actin-accumulation myopathy

    MedlinePlus

    ... 7(3):160-8. Citation on PubMed Laing NG, Dye DE, Wallgren-Pettersson C, Richard G, Monnier ... Vigneron J, Wallgren-Pettersson C, Beggs AH, Laing NG. Mutations in the skeletal muscle alpha-actin gene ...

  1. Actin expression in trypanosomatids (Euglenozoa: Kinetoplastea).

    PubMed

    Souza, Ligia Cristina Kalb; Pinho, Rosana Elisa Gonçalves Gonçalves; Lima, Carla Vanessa de Paula; Fragoso, Stênio Perdigão; Soares, Maurilio José

    2013-08-01

    Heteroxenic and monoxenic trypanosomatids were screened for the presence of actin using a mouse polyclonal antibody produced against the entire sequence of the Trypanosoma cruzi actin gene, encoding a 41.9 kDa protein. Western blot analysis showed that this antibody reacted with a polypeptide of approximately 42 kDa in the whole-cell lysates of parasites targeting mammals (T. cruzi, Trypanosoma brucei and Leishmania major), insects (Angomonas deanei, Crithidia fasciculata, Herpetomonas samuelpessoai and Strigomonas culicis) and plants (Phytomonas serpens). A single polypeptide of approximately 42 kDa was detected in the whole-cell lysates of T. cruzi cultured epimastigotes, metacyclic trypomastigotes and amastigotes at similar protein expression levels. Confocal microscopy showed that actin was expressed throughout the cytoplasm of all the tested trypanosomatids. These data demonstrate that actin expression is widespread in trypanosomatids. PMID:23903980

  2. Actin expression in trypanosomatids (Euglenozoa: Kinetoplastea)

    PubMed Central

    Souza, Ligia Cristina Kalb; Pinho, Rosana Elisa Gonçalves Gonçalves; Lima, Carla Vanessa de Paula; Fragoso, Stênio Perdigão; Soares, Maurilio José

    2013-01-01

    Heteroxenic and monoxenic trypanosomatids were screened for the presence of actin using a mouse polyclonal antibody produced against the entire sequence of the Trypanosoma cruzi actin gene, encoding a 41.9 kDa protein. Western blot analysis showed that this antibody reacted with a polypeptide of approximately 42 kDa in the whole-cell lysates of parasites targeting mammals (T. cruzi, Trypanosoma brucei and Leishmania major), insects (Angomonas deanei, Crithidia fasciculata, Herpetomonas samuelpessoai and Strigomonas culicis) and plants (Phytomonas serpens). A single polypeptide of approximately 42 kDa was detected in the whole-cell lysates of T. cruzi cultured epimastigotes, metacyclic trypomastigotes and amastigotes at similar protein expression levels. Confocal microscopy showed that actin was expressed throughout the cytoplasm of all the tested trypanosomatids. These data demonstrate that actin expression is widespread in trypanosomatids. PMID:23903980

  3. [Actin in the wound healing process].

    PubMed

    Nowak, Dorota; Popow-Woźniak, Agnieszka; Raźnikiewicz, Linda; Malicka-Błaszkiewicz, Maria

    2009-01-01

    Wound healing is an important biological process of crucial value for organisms survival and retention of its proper functions. The recognition of molecular mechanisms of these phenomenon is still under investigation. The transition of mesenchymal fibroblasts to myofibroblasts is a key point in wound healing. The contraction ability of myofibroblast enables the shrinkage of a wound and closes its edges. Alpha smooth muscle actin (alpha-SMA), one of six actin isoforms, is a marker of compeletely differentiated myofibroblast. The regulation of differentiation process depends on many growth factors (especially TGF beta 1), the level of active thymosin beta 4, extracellular matrix proteins--including fibronectin, and also on specificity of microenvironment. Thymosin beta 4 is responsible for maintenance of pool of monomeric actin and actin filaments depolymerization. It can also act as a transcription factor, migration stimulator and immunomodulator, so this protein deserves for more attention in wound healing research field. PMID:19824469

  4. Mechanics model for actin-based motility

    NASA Astrophysics Data System (ADS)

    Lin, Yuan

    2009-02-01

    We present here a mechanics model for the force generation by actin polymerization. The possible adhesions between the actin filaments and the load surface, as well as the nucleation and capping of filament tips, are included in this model on top of the well-known elastic Brownian ratchet formulation. A closed form solution is provided from which the force-velocity relationship, summarizing the mechanics of polymerization, can be drawn. Model predictions on the velocity of moving beads driven by actin polymerization are consistent with experiment observations. This model also seems capable of explaining the enhanced actin-based motility of Listeria monocytogenes and beads by the presence of Vasodilator-stimulated phosphoprotein, as observed in recent experiments.

  5. Structural dynamics of an actin spring.

    PubMed

    Mahadevan, L; Riera, C S; Shin, Jennifer H

    2011-02-16

    Actin-based motility in cells is usually associated with either polymerization/depolymerization in the presence of cross-linkers or contractility in the presence of myosin motors. Here, we focus on a third distinct mechanism involving actin in motility, seen in the dynamics of an active actin spring that powers the acrosomal reaction of the horseshoe crab (Limulus polyphemus) sperm. During this process, a 60-μm bent and twisted bundle of cross-linked actin uncoils and becomes straight in a few seconds in the presence of Ca(2+). This straightening, which occurs at a constant velocity, allows the acrosome to forcefully penetrate the egg. Synthesizing ultrastructural information with the kinetics, energetics, and imaging of calcium binding allows us to construct a dynamical theory for this mechanochemical engine consistent with our experimental observations. It also illuminates the general mechanism by which energy may be stored in conformational changes and released cooperatively in ordered macromolecular assemblies. PMID:21320427

  6. A microstructurally informed model for the mechanical response of three-dimensional actin networks

    PubMed Central

    KWON, R.Y.; LEW, A.J.; JACOBS, C.R.

    2008-01-01

    We propose a class of microstructurally informed models for the linear elastic mechanical behavior of cross-linked polymer networks such as the actin cytoskeleton. Salient features of the models include the possibility to represent anisotropic mechanical behavior resulting from anisotropic filament distributions, and a power-law scaling of the mechanical properties with the filament density. Mechanical models within the class are parameterized by seven different constants. We demonstrate a procedure for determining these constants using finite element models of three-dimensional actin networks. Actin filaments and cross-links were modeled as elastic rods, and the networks were constructed at physiological volume fractions and at the scale of an image voxel. We show the performance of the model in estimating the mechanical behavior of the networks over a wide range of filament densities and degrees of anisotropy. PMID:18568835

  7. Actinic review of EUV masks

    NASA Astrophysics Data System (ADS)

    Feldmann, Heiko; Ruoff, Johannes; Harnisch, Wolfgang; Kaiser, Winfried

    2010-04-01

    Management of mask defects is a major challenge for the introduction of EUV for HVM production. Once a defect has been detected, its printing impact needs to be predicted. Potentially the defect requires some repair, the success of which needs to be proven. This defect review has to be done with an actinic inspection system that matches the imaging conditions of an EUV scanner. During recent years, several concepts for such an aerial image metrology system (AIMS™) have been proposed. However, until now no commercial solution exists for EUV. Today, advances in EUV optics technology allow envisioning a solution that has been discarded before as unrealistic. We present this concept and its technical cornerstones.While the power requirement for the EUV source is less demanding than for HVM lithography tools, radiance, floor space, and stability are the main criteria for source selection. The requirement to emulate several generations of EUV scanners demands a large flexibility for the ilumination and imaging systems. New critical specifications to the EUV mirrors in the projection microscope can be satisfied using our expertise from lithographic mirrors. In summary, an EUV AIMS™ meeting production requirements seems to be feasible.

  8. The actin cytoskeleton in presynaptic assembly.

    PubMed

    Nelson, Jessica C; Stavoe, Andrea K H; Colón-Ramos, Daniel A

    2013-01-01

    Dramatic morphogenetic processes underpin nearly every step of nervous system development, from initial neuronal migration and axon guidance to synaptogenesis. Underlying this morphogenesis are dynamic rearrangements of cytoskeletal architecture. Here we discuss the roles of the actin cytoskeleton in the development of presynaptic terminals, from the elaboration of terminal arbors to the recruitment of presynaptic vesicles and active zone components. The studies discussed here underscore the importance of actin regulation at every step in neuronal circuit assembly. PMID:23628914

  9. Mechanism of Actin Filament Bundling by Fascin

    SciTech Connect

    Jansen, Silvia; Collins, Agnieszka; Yang, Changsong; Rebowski, Grzegorz; Svitkina, Tatyana; Dominguez, Roberto

    2013-03-07

    Fascin is the main actin filament bundling protein in filopodia. Because of the important role filopodia play in cell migration, fascin is emerging as a major target for cancer drug discovery. However, an understanding of the mechanism of bundle formation by fascin is critically lacking. Fascin consists of four {beta}-trefoil domains. Here, we show that fascin contains two major actin-binding sites, coinciding with regions of high sequence conservation in {beta}-trefoil domains 1 and 3. The site in {beta}-trefoil-1 is located near the binding site of the fascin inhibitor macroketone and comprises residue Ser-39, whose phosphorylation by protein kinase C down-regulates actin bundling and formation of filopodia. The site in {beta}-trefoil-3 is related by pseudo-2-fold symmetry to that in {beta}-trefoil-1. The two sites are {approx}5 nm apart, resulting in a distance between actin filaments in the bundle of {approx}8.1 nm. Residue mutations in both sites disrupt bundle formation in vitro as assessed by co-sedimentation with actin and electron microscopy and severely impair formation of filopodia in cells as determined by rescue experiments in fascin-depleted cells. Mutations of other areas of the fascin surface also affect actin bundling and formation of filopodia albeit to a lesser extent, suggesting that, in addition to the two major actin-binding sites, fascin makes secondary contacts with other filaments in the bundle. In a high resolution crystal structure of fascin, molecules of glycerol and polyethylene glycol are bound in pockets located within the two major actin-binding sites. These molecules could guide the rational design of new anticancer fascin inhibitors.

  10. Actin filament curvature biases branching direction

    NASA Astrophysics Data System (ADS)

    Wang, Evan; Risca, Viviana; Chaudhuri, Ovijit; Chia, Jia-Jun; Geissler, Phillip; Fletcher, Daniel

    2012-02-01

    Actin filaments are key components of the cellular machinery, vital for a wide range of processes ranging from cell motility to endocytosis. Actin filaments can branch, and essential in this process is a protein complex known as the Arp2/3 complex, which nucleate new ``daughter'' filaments from pre-existing ``mother'' filaments by attaching itself to the mother filament. Though much progress has been made in understanding the Arp2/3-actin junction, some very interesting questions remain. In particular, F-actin is a dynamic polymer that undergoes a wide range of fluctuations. Prior studies of the Arp2/3-actin junction provides a very static notion of Arp2/3 binding. The question we ask is how differently does the Arp2/3 complex interact with a straight filament compared to a bent filament? In this study, we used Monte Carlo simulations of a surface-tethered worm-like chain to explore possible mechanisms underlying the experimental observation that there exists preferential branch formation by the Arp2/3 complex on the convex face of a curved filament. We show that a fluctuation gating model in which Arp2/3 binding to the actin filament is dependent upon a rare high-local-curvature shape fluctuation of the filament is consistent with the experimental data.

  11. The Bacterial Actin-Like Cytoskeleton

    PubMed Central

    Carballido-López, Rut

    2006-01-01

    Recent advances have shown conclusively that bacterial cells possess distant but true homologues of actin (MreB, ParM, and the recently uncovered MamK protein). Despite weak amino acid sequence similarity, MreB and ParM exhibit high structural homology to actin. Just like F-actin in eukaryotes, MreB and ParM assemble into highly dynamic filamentous structures in vivo and in vitro. MreB-like proteins are essential for cell viability and have been implicated in major cellular processes, including cell morphogenesis, chromosome segregation, and cell polarity. ParM (a plasmid-encoded actin homologue) is responsible for driving plasmid-DNA partitioning. The dynamic prokaryotic actin-like cytoskeleton is thought to serve as a central organizer for the targeting and accurate positioning of proteins and nucleoprotein complexes, thereby (and by analogy to the eukaryotic cytoskeleton) spatially and temporally controlling macromolecular trafficking in bacterial cells. In this paper, the general properties and known functions of the actin orthologues in bacteria are reviewed. PMID:17158703

  12. Structure of a Longitudinal Actin Dimer Assembled by Tandem W Domains: Implications for Actin Filament Nucleation

    SciTech Connect

    Rebowski, Grzegorz; Namgoong, Suk; Boczkowska, Malgorzata; Leavis, Paul C.; Navaza, Jorge; Dominguez, Roberto

    2013-11-20

    Actin filament nucleators initiate polymerization in cells in a regulated manner. A common architecture among these molecules consists of tandem WASP homology 2 domains (W domains) that recruit three to four actin subunits to form a polymerization nucleus. We describe a low-resolution crystal structure of an actin dimer assembled by tandem W domains, where the first W domain is cross-linked to Cys374 of the actin subunit bound to it, whereas the last W domain is followed by the C-terminal pointed end-capping helix of thymosin {beta}4. While the arrangement of actin subunits in the dimer resembles that of a long-pitch helix of the actin filament, important differences are observed. These differences result from steric hindrance of the W domain with intersubunit contacts in the actin filament. We also determined the structure of the first W domain of Vibrio parahaemolyticus VopL cross-linked to actin Cys374 and show it to be nearly identical with non-cross-linked W-Actin structures. This result validates the use of cross-linking as a tool for the study of actin nucleation complexes, whose natural tendency to polymerize interferes with most structural methods. Combined with a biochemical analysis of nucleation, the structures may explain why nucleators based on tandem W domains with short inter-W linkers have relatively weak activity, cannot stay bound to filaments after nucleation, and are unlikely to influence filament elongation. The findings may also explain why nucleation-promoting factors of the Arp2/3 complex, which are related to tandem-W-domain nucleators, are ejected from branch junctions after nucleation. We finally show that the simple addition of the C-terminal pointed end-capping helix of thymosin {beta}4 to tandem W domains can change their activity from actin filament nucleation to monomer sequestration.

  13. Mechanical Heterogeneity Favors Fragmentation of Strained Actin Filaments

    PubMed Central

    De La Cruz, Enrique M.; Martiel, Jean-Louis; Blanchoin, Laurent

    2015-01-01

    We present a general model of actin filament deformation and fragmentation in response to compressive forces. The elastic free energy density along filaments is determined by their shape and mechanical properties, which were modeled in terms of bending, twisting, and twist-bend coupling elasticities. The elastic energy stored in filament deformation (i.e., strain) tilts the fragmentation-annealing reaction free-energy profile to favor fragmentation. The energy gradient introduces a local shear force that accelerates filament intersubunit bond rupture. The severing protein, cofilin, renders filaments more compliant in bending and twisting. As a result, filaments that are partially decorated with cofilin are mechanically heterogeneous (i.e., nonuniform) and display asymmetric shape deformations and energy profiles distinct from mechanically homogenous (i.e., uniform), bare actin, or saturated cofilactin filaments. The local buckling strain depends on the relative size of the compliant segment as well as the bending and twisting rigidities of flanking regions. Filaments with a single bare/cofilin-decorated boundary localize energy and force adjacent to the boundary, within the compliant cofilactin segment. Filaments with small cofilin clusters were predicted to fragment within the compliant cofilactin rather than at boundaries. Neglecting contributions from twist-bend coupling elasticity underestimates the energy density and gradients along filaments, and thus the net effects of filament strain to fragmentation. Spatial confinement causes compliant cofilactin segments and filaments to adopt higher deformation modes and store more elastic energy, thereby promoting fragmentation. The theory and simulations presented here establish a quantitative relationship between actin filament fragmentation thermodynamics and elasticity, and reveal how local discontinuities in filament mechanical properties introduced by regulatory proteins can modulate both the severing efficiency

  14. Critical current density, irreversibility line, and flux creep activation energy in silver-sheathed Bi[sub 2]Sr[sub 2]Ca[sub 2]Cu[sub 3]O[sub x] superconducting tapes

    SciTech Connect

    Shi, D.; Wang, Z.; Sengupta, S.; Smith, M. ); Goodrich, L.F. , Boulder, CO . Electromagnetic Technology Div.); Dou, S.X.; Liu, H.K.; Guo, Y.C. . School of Materials and Engineering)

    1992-08-01

    Transport data, magnetic hysteresis and flux creep activation energy experimental results are presented for silver-sheathed high-[Tc] Bi[sub 2]Sr[sub 2]Ca[sub 2]Cu[sub 3]O[sub x] superconducting tapes. The 110 K superconducting phase was formed by lead doping in a Bi-Sr-Ca-Cu-0 system. The transport critical current density was measured at 4.0 K to be 0.7 [times] 10[sup 5] A/cm[sup 2] (the corresponding critical current is 74 A) at zero field and 1.6 [times] 10[sup 4] A/cm[sup 2] at 12 T for H[parallel]ab. Excellent grain alignment in the a-b plane was achieved by a short-melting method, which considerably improved the critical current density and irreversibility line. Flux creep activation energy as a function of current is obtained based on the magnetic relaxation measurements.

  15. Critical current density, irreversibility line, and flux creep activation energy in silver-sheathed Bi{sub 2}Sr{sub 2}Ca{sub 2}Cu{sub 3}O{sub x} superconducting tapes

    SciTech Connect

    Shi, D.; Wang, Z.; Sengupta, S.; Smith, M.; Goodrich, L.F.; Dou, S.X.; Liu, H.K.; Guo, Y.C.

    1992-08-01

    Transport data, magnetic hysteresis and flux creep activation energy experimental results are presented for silver-sheathed high-{Tc} Bi{sub 2}Sr{sub 2}Ca{sub 2}Cu{sub 3}O{sub x} superconducting tapes. The 110 K superconducting phase was formed by lead doping in a Bi-Sr-Ca-Cu-0 system. The transport critical current density was measured at 4.0 K to be 0.7 {times} 10{sup 5} A/cm{sup 2} (the corresponding critical current is 74 A) at zero field and 1.6 {times} 10{sup 4} A/cm{sup 2} at 12 T for H{parallel}ab. Excellent grain alignment in the a-b plane was achieved by a short-melting method, which considerably improved the critical current density and irreversibility line. Flux creep activation energy as a function of current is obtained based on the magnetic relaxation measurements.

  16. Cofilin-induced cooperative conformational changes of actin subunits revealed using cofilin-actin fusion protein

    PubMed Central

    Umeki, Nobuhisa; Hirose, Keiko; Uyeda, Taro Q. P.

    2016-01-01

    To investigate cooperative conformational changes of actin filaments induced by cofilin binding, we engineered a fusion protein made of Dictyostelium cofilin and actin. The filaments of the fusion protein were functionally similar to actin filaments bound with cofilin in that they did not bind rhodamine-phalloidin, had quenched fluorescence of pyrene attached to Cys374 and showed enhanced susceptibility of the DNase loop to cleavage by subtilisin. Quantitative analyses of copolymers made of different ratios of the fusion protein and control actin further demonstrated that the fusion protein affects the structure of multiple neighboring actin subunits in copolymers. Based on these and other recent related studies, we propose a mechanism by which conformational changes induced by cofilin binding is propagated unidirectionally to the pointed ends of the filaments, and cofilin clusters grow unidirectionally to the pointed ends following this path. Interestingly, the fusion protein was unable to copolymerize with control actin at pH 6.5 and low ionic strength, suggesting that the structural difference between the actin moiety in the fusion protein and control actin is pH-sensitive. PMID:26842224

  17. Tau co-organizes dynamic microtubule and actin networks

    PubMed Central

    Elie, Auréliane; Prezel, Elea; Guérin, Christophe; Denarier, Eric; Ramirez-Rios, Sacnicte; Serre, Laurence; Andrieux, Annie; Fourest-Lieuvin, Anne; Blanchoin, Laurent; Arnal, Isabelle

    2015-01-01

    The crosstalk between microtubules and actin is essential for cellular functions. However, mechanisms underlying the microtubule-actin organization by cross-linkers remain largely unexplored. Here, we report that tau, a neuronal microtubule-associated protein, binds to microtubules and actin simultaneously, promoting in vitro co-organization and coupled growth of both networks. By developing an original assay to visualize concomitant microtubule and actin assembly, we show that tau can induce guided polymerization of actin filaments along microtubule tracks and growth of single microtubules along actin filament bundles. Importantly, tau mediates microtubule-actin co-alignment without changing polymer growth properties. Mutagenesis studies further reveal that at least two of the four tau repeated motifs, primarily identified as tubulin-binding sites, are required to connect microtubules and actin. Tau thus represents a molecular linker between microtubule and actin networks, enabling a coordination of the two cytoskeletons that might be essential in various neuronal contexts. PMID:25944224

  18. Structural Basis of Actin Filament Nucleation by Tandem W Domains

    PubMed Central

    Chen, Xiaorui; Ni, Fengyun; Tian, Xia; Kondrashkina, Elena; Wang, Qinghua; Ma, Jianpeng

    2013-01-01

    SUMMARY Spontaneous nucleation of actin is very inefficient in cells. To overcome this barrier, cells have evolved a set of actin filament nucleators to promote rapid nucleation and polymerization in response to specific stimuli. However, the molecular mechanism of actin nucleation remains poorly understood. This is hindered largely by the fact that actin nucleus, once formed, rapidly polymerizes into filament, thus making it impossible to capture stable multisubunit actin nucleus. Here, we report an effective double-mutant strategy to stabilize actin nucleus by preventing further polymerization. Employing this strategy, we solved the crystal structure of AMPPNP-actin in complex with the first two tandem W domains of Cordon-bleu (Cobl), a potent actin filament nucleator. Further sequence comparison and functional studies suggest that the nucleation mechanism of Cobl is probably shared by the p53 cofactor JMY, but not Spire. Moreover, the double-mutant strategy opens the way for atomic mechanistic study of actin nucleation and polymerization. PMID:23727244

  19. Sensing actin dynamics: Structural basis for G-actin-sensitive nuclear import of MAL

    SciTech Connect

    Hirano, Hidemi; Matsuura, Yoshiyuki

    2011-10-22

    Highlights: {yields} MAL has a bipartite NLS that binds to Imp{alpha} in an extended conformation. {yields} Mutational analyses verified the functional significance of MAL-Imp{alpha} interactions. {yields} Induced folding and NLS-masking by G-actins inhibit nuclear import of MAL. -- Abstract: The coordination of cytoskeletal actin dynamics with gene expression reprogramming is emerging as a crucial mechanism to control diverse cellular processes, including cell migration, differentiation and neuronal circuit assembly. The actin-binding transcriptional coactivator MAL (also known as MRTF-A/MKL1/BSAC) senses G-actin concentration and transduces Rho GTPase signals to serum response factor (SRF). MAL rapidly shuttles between the cytoplasm and the nucleus in unstimulated cells but Rho-induced depletion of G-actin leads to MAL nuclear accumulation and activation of transcription of SRF:MAL-target genes. Although the molecular and structural basis of actin-regulated nucleocytoplasmic shuttling of MAL is not understood fully, it is proposed that nuclear import of MAL is mediated by importin {alpha}/{beta} heterodimer, and that G-actin competes with importin {alpha}/{beta} for the binding to MAL. Here we present structural, biochemical and cell biological evidence that MAL has a classical bipartite nuclear localization signal (NLS) in the N-terminal 'RPEL' domain containing Arg-Pro-X-X-X-Glu-Leu (RPEL) motifs. The NLS residues of MAL adopt an extended conformation and bind along the surface groove of importin-{alpha}, interacting with the major- and minor-NLS binding sites. We also present a crystal structure of wild-type MAL RPEL domain in complex with five G-actins. Comparison of the importin-{alpha}- and actin-complexes revealed that the binding of G-actins to MAL is associated with folding of NLS residues into a helical conformation that is inappropriate for importin-{alpha} recognition.

  20. Glutamyl Phosphate Is an Activated Intermediate in Actin Crosslinking by Actin Crosslinking Domain (ACD) Toxin

    PubMed Central

    Kudryashova, Elena; Kalda, Caitlin; Kudryashov, Dmitri S.

    2012-01-01

    Actin Crosslinking Domain (ACD) is produced by several life-threatening Gram-negative pathogenic bacteria as part of larger toxins and delivered into the cytoplasm of eukaryotic host cells via Type I or Type VI secretion systems. Upon delivery, ACD disrupts the actin cytoskeleton by catalyzing intermolecular amide bond formation between E270 and K50 residues of actin, leading to the formation of polymerization-deficient actin oligomers. Ultimately, accumulation of the crosslinked oligomers results in structural and functional failure of the actin cytoskeleton in affected cells. In the present work, we advanced in our understanding of the ACD catalytic mechanism by discovering that the enzyme transfers the gamma-phosphoryl group of ATP to the E270 actin residue, resulting in the formation of an activated acyl phosphate intermediate. This intermediate is further hydrolyzed and the energy of hydrolysis is utilized for the formation of the amide bond between actin subunits. We also determined the pH optimum for the reaction and the kinetic parameters of ACD catalysis for its substrates, ATP and actin. ACD showed sigmoidal, non-Michaelis-Menten kinetics for actin (K0.5 = 30 µM) reflecting involvement of two actin molecules in a single crosslinking event. We established that ACD can also utilize Mg2+-GTP to support crosslinking, but the kinetic parameters (KM = 8 µM and 50 µM for ATP and GTP, respectively) suggest that ATP is the primary substrate of ACD in vivo. The optimal pH for ACD activity was in the range of 7.0–9.0. The elucidated kinetic mechanism of ACD toxicity adds to understanding of complex network of host-pathogen interactions. PMID:23029200

  1. The natural product cucurbitacin E inhibits depolymerization of actin filaments

    PubMed Central

    Sörensen, Pia M.; Iacob, Roxana E.; Fritzsche, Marco; Engen, John R.; Brieher, William M.; Charras, Guillaume; Eggert, Ulrike S.

    2012-01-01

    Although small molecule actin modulators have been widely used as research tools, only one cell permeable small molecule inhibitor of actin depolymerization (jasplakinolide) is commercially available. We report that the natural product cucurbitacin E inhibits actin depolymerization and show that its mechanism of action is different from jasplakinolide. In assays using pure fluorescently labeled actin, cucurbitacin E specifically affected depolymerization without affecting polymerization. It inhibited actin depolymerization at sub-stoichiometric concentrations up to 1:6 cucurbitacin:actin E. Cucurbitacin E specifically binds to filamentous actin (F-actin) forming a covalent bond at residue Cys257, but not to monomeric actin (G-actin). Based on its compatibility with phalloidin staining, we show that cucurbitacin E occupies a different binding site on actin filaments. Using loss of fluorescence after localized photoactivation, we found that cucurbitacin E inhibited actin depolymerization in live cells. Cucurbitacin E is a widely available plant-derived natural product, making it a useful tool to study actin dynamics in cells and actin-based processes such as cytokinesis. PMID:22724897

  2. Incorporation of mammalian actin into microfilaments in plant cell nucleus

    PubMed Central

    Paves, Heiti; Truve, Erkki

    2004-01-01

    Background Actin is an ancient molecule that shows more than 90% amino acid homology between mammalian and plant actins. The regions of the actin molecule that are involved in F-actin assembly are largely conserved, and it is likely that mammalian actin is able to incorporate into microfilaments in plant cells but there is no experimental evidence until now. Results Visualization of microfilaments in onion bulb scale epidermis cells by different techniques revealed that rhodamine-phalloidin stained F-actin besides cytoplasm also in the nuclei whereas GFP-mouse talin hybrid protein did not enter the nuclei. Microinjection of fluorescently labeled actin was applied to study the presence of nuclear microfilaments in plant cells. Ratio imaging of injected fluorescent rabbit skeletal muscle actin and phalloidin staining of the microinjected cells showed that mammalian actin was able to incorporate into plant F-actin. The incorporation occurred preferentially in the nucleus and in the perinuclear region of plant cells whereas part of plant microfilaments, mostly in the periphery of cytoplasm, did not incorporate mammalian actin. Conclusions Microinjected mammalian actin is able to enter plant cell's nucleus, whereas incorporation of mammalian actin into plant F-actin occurs preferentially in the nucleus and perinuclear area. PMID:15102327

  3. Actin-dependent mechanisms in AMPA receptor trafficking

    PubMed Central

    Hanley, Jonathan G.

    2014-01-01

    The precise regulation of AMPA receptor (AMPAR) number and subtype at the synapse is crucial for the regulation of excitatory neurotransmission, synaptic plasticity and the consequent formation of appropriate neural circuits for learning and memory. AMPAR trafficking involves the dynamic processes of exocytosis, endocytosis and endosomal recycling, all of which involve the actin cytoskeleton. The actin cytoskeleton is highly dynamic and highly regulated by an abundance of actin-binding proteins and upstream signaling pathways that modulate actin polymerization and depolymerization. Actin dynamics generate forces that manipulate membranes in the process of vesicle biogenesis, and also for propelling vesicles through the cytoplasm to reach their destination. In addition, trafficking mechanisms exploit more stable aspects of the actin cytoskeleton by using actin-based motor proteins to traffic vesicular cargo along actin filaments. Numerous studies have shown that actin dynamics are critical for AMPAR localization and function. The identification of actin-binding proteins that physically interact with AMPAR subunits, and research into their mode of action is starting to shed light on the mechanisms involved. Such proteins either regulate actin dynamics to modulate mechanical forces exerted on AMPAR-containing membranes, or associate with actin filaments to target or transport AMPAR-containing vesicles to specific subcellular regions. In addition, actin-regulatory proteins that do not physically interact with AMPARs may influence AMPAR trafficking by regulating the local actin environment in the dendritic spine. PMID:25429259

  4. Crystal structure of a nuclear actin ternary complex.

    PubMed

    Cao, Tingting; Sun, Lingfei; Jiang, Yuxiang; Huang, Shanjin; Wang, Jiawei; Chen, Zhucheng

    2016-08-01

    Actin polymerizes and forms filamentous structures (F-actin) in the cytoplasm of eukaryotic cells. It also exists in the nucleus and regulates various nucleic acid transactions, particularly through its incorporation into multiple chromatin-remodeling complexes. However, the specific structure of actin and the mechanisms that regulate its polymeric nature inside the nucleus remain unknown. Here, we report the crystal structure of nuclear actin (N-actin) complexed with actin-related protein 4 (Arp4) and the helicase-SANT-associated (HSA) domain of the chromatin remodeler Swr1. The inner face and barbed end of N-actin are sequestered by interactions with Arp4 and the HSA domain, respectively, which prevents N-actin from polymerization and binding to many actin regulators. The two major domains of N-actin are more twisted than those of globular actin (G-actin), and its nucleotide-binding pocket is occluded, freeing N-actin from binding to and regulation by ATP. These findings revealed the salient structural features of N-actin that distinguish it from its cytoplasmic counterpart and provide a rational basis for its functions and regulation inside the nucleus. PMID:27457955

  5. Supercoiling of f-actin filaments.

    PubMed

    Lednev, V V; Popp, D

    1990-05-01

    In the X-ray diffraction pattern from oriented gels of actin-containing filaments sampling of layer lines indicating the development of a well-ordered pseudo-hexagonal lattice within the gels at interfilament spacings as large as 13 nm is observed. This value exceeds by 3 nm the largest estimate of an external diameter of pure f-actin. The development of layer line sampling is always accompanied by: (i) the appearance of strong forbidden meridional reflections on the 5.9- and 5.1-nm layer lines; (ii) a drastic intensification of the first (expected) 2.75-nm meridional reflection by a factor of about 4; (iii) the appearance of streaks, connecting near-meridional reflections on the 5.9-, 5.1-, and 37-nm layer lines; and (iv) a slight decrease in the number of subunits per turn of the basic f-actin helix. All these features strongly indicate that f-actin filaments are supercoiled and make regular local contacts between themselves, which may lead to periodic distortions of the mobile external domain in the actin subunits. PMID:2261308

  6. Impact of Carbon Nanomaterials on Actin Polymerization.

    PubMed

    Dong, Ying; Sun, Haiyan; Li, Xu; Li, Xin; Zhao, Lina

    2016-03-01

    Many nanomaterials have entered people's daily lives and impact the normal process of biological entities consequently. As one kind of the important nanomaterials, carbon based nanomaterials have invoked a lot of concerns from scientific researches because of their unique physicochemical properties. In eukaryotes, actin is the most abundantly distributed protein in both cytoplasm and cell nucleus, and closely controls the cell proliferation and mobility. Recently, many investigations have found some carbon based nanomaterials can affect actin cytoskeleton remarkably, including fullerenes derivatives, carbon nanotubes, graphene and its derivatives. However, these interaction processes are complicated and the underlying mechanism is far from being understood clearly. In this review, we discussed the different mechanisms of carbon nanomaterials impact on actin polymerization into three pathways, as triggering the signaling pathways from carbon nanomaterials outside of cells, increasing the production of reactive oxygen species from carbon nanomaterials inside of cells and direct interaction from carbon nanomaterials inside of cells. As a result, the dimension and size of carbon nanomaterials play a key role in regulation of actin cytoskeleton. Furthermore, we forecasted the possible investigation strategy for meeting the challenges of the future study on this topic. We hope the findings are helpful in understanding the molecular mechanism in carbon nanomaterials regulating actin polymerization, and provide new insight in novel nanomedicine development for inhibition tumor cell migration. PMID:27455649

  7. Structural Transitions of F-Actin:Espin Bundles

    NASA Astrophysics Data System (ADS)

    Purdy, Kirstin; Bartles, James; Wong, Gerard

    2006-03-01

    Espin is an actin bundling protein involved in the formation of the parallel bundles of filamentous actin in hair cell stereocilia. Mutations in espin are implicated in deafness phenotypes in mice and humans. We present measurements of the F-actin structures induced by wild type and by mutated espin obtained via small angle x-ray scattering and fluorescence microscopy. We found that wild type espin induced a paracrystalline hexagonal array of twisted F-actin, whereas the mutated espin only condensed the F-actin into a nematic-like phase. The possibility of coexisting nematic and bundled actin in mixtures containing both mutant and wild type espins was also investigated.

  8. Actin Filament Segmentation Using Dynamic Programming

    PubMed Central

    Li, Hongsheng; Shen, Tian; Huang, Xiaolei

    2011-01-01

    We introduce a novel algorithm for actin filament segmentation in 2D TIRFM image sequences. This problem is difficult because actin filaments dynamically change shapes during their growth, and the TIRFM images are usually noisy. We ask a user to specify the two tips of a filament of interest in the first frame. We then model the segmentation problem in an image sequence as a temporal chain, where its states are tip locations; given candidate tip locations, actin filaments' body points are inferred by a dynamic programming method, which adaptively generates candidate solutions. Combining candidate tip locations and their inferred body points, the temporal chain model is efficiently optimized using another dynamic programming method. Evaluation on noisy TIRFM image sequences demonstrates the accuracy and robustness of this approach. PMID:21761674

  9. Ionic wave propagation along actin filaments.

    PubMed

    Tuszyński, J A; Portet, S; Dixon, J M; Luxford, C; Cantiello, H F

    2004-04-01

    We investigate the conditions enabling actin filaments to act as electrical transmission lines for ion flows along their lengths. We propose a model in which each actin monomer is an electric element with a capacitive, inductive, and resistive property due to the molecular structure of the actin filament and viscosity of the solution. Based on Kirchhoff's laws taken in the continuum limit, a nonlinear partial differential equation is derived for the propagation of ionic waves. We solve this equation in two different regimes. In the first, the maximum propagation velocity wave is found in terms of Jacobi elliptic functions. In the general case, we analyze the equation in terms of Fisher-Kolmogoroff modes with both localized and extended wave characteristics. We propose a new signaling mechanism in the cell, especially in neurons. PMID:15041636

  10. Spontaneous actin dynamics in contractile rings

    NASA Astrophysics Data System (ADS)

    Kruse, Karsten; Wollrab, Viktoria; Thiagarajan, Raghavan; Wald, Anne; Riveline, Daniel

    Networks of polymerizing actin filaments are known to be capable to self-organize into a variety of structures. For example, spontaneous actin polymerization waves have been observed in living cells in a number of circumstances, notably, in crawling neutrophils and slime molds. During later stages of cell division, they can also spontaneously form a contractile ring that will eventually cleave the cell into two daughter cells. We present a framework for describing networks of polymerizing actin filaments, where assembly is regulated by various proteins. It can also include the effects of molecular motors. We show that the molecular processes driven by these proteins can generate various structures that have been observed in contractile rings of fission yeast and mammalian cells. We discuss a possible functional role of each of these patterns. The work was supported by Agence Nationale de la Recherche, France, (ANR-10-LABX-0030-INRT) and by Deutsche Forschungsgemeinschaft through SFB1027.

  11. Myosin VI small insert isoform maintains exocytosis by tethering secretory granules to the cortical actin

    PubMed Central

    Tomatis, Vanesa M.; Papadopulos, Andreas; Malintan, Nancy T.; Martin, Sally; Wallis, Tristan; Gormal, Rachel S.; Kendrick-Jones, John; Buss, Folma

    2013-01-01

    Before undergoing neuroexocytosis, secretory granules (SGs) are mobilized and tethered to the cortical actin network by an unknown mechanism. Using an SG pull-down assay and mass spectrometry, we found that myosin VI was recruited to SGs in a Ca2+-dependent manner. Interfering with myosin VI function in PC12 cells reduced the density of SGs near the plasma membrane without affecting their biogenesis. Myosin VI knockdown selectively impaired a late phase of exocytosis, consistent with a replenishment defect. This exocytic defect was selectively rescued by expression of the myosin VI small insert (SI) isoform, which efficiently tethered SGs to the cortical actin network. These myosin VI SI–specific effects were prevented by deletion of a c-Src kinase phosphorylation DYD motif, identified in silico. Myosin VI SI thus recruits SGs to the cortical actin network, potentially via c-Src phosphorylation, thereby maintaining an active pool of SGs near the plasma membrane. PMID:23382463

  12. The actin binding protein adseverin regulates osteoclastogenesis.

    PubMed

    Hassanpour, Siavash; Jiang, Hongwei; Wang, Yongqiang; Kuiper, Johannes W P; Glogauer, Michael

    2014-01-01

    Adseverin (Ads), a member of the Gelsolin superfamily of actin binding proteins, regulates the actin cytoskeleton architecture by severing and capping existing filamentous actin (F-actin) strands and nucleating the assembly of new F-actin filaments. Ads has been implicated in cellular secretion, exocytosis and has also been shown to regulate chondrogenesis and megakaryoblastic leukemia cell differentiation. Here we report for the first time that Ads is involved in regulating osteoclastogenesis (OCG). Ads is induced during OCG downstream of RANK-ligand (RANKL) stimulation and is highly expressed in mature osteoclasts. The D5 isoform of Ads is not involved in regulating OCG, as its expression is not induced in response to RANKL. Three clonal Ads knockdown RAW264.7 (RAW) macrophage cell lines with varying degrees of Ads expression and OCG deficiency were generated. The most drastic OCG defect was noted in the clonal cell line with the greatest degree of Ads knockdown as indicated by a lack of TRAcP staining and multinucleation. RNAi mediated knockdown of Ads in osteoclast precursors resulted in distinct morphological changes characterized by altered F-actin distribution and increased filopodia formation. Ads knockdown precursor cells experienced enhanced migration while fusion of knockdown precursors cells was limited. Transient reintroduction of de novo Ads back into the knockdown system was capable of rescuing TRAcP expression but not osteoclast multinucleation most likely due to the transient nature of Ads expression. This preliminary study allows us to conclude that Ads is a RANKL induced early regulator of OCG with a potential role in pre-osteoclast differentiation and fusion. PMID:25275604

  13. The Actin Binding Protein Adseverin Regulates Osteoclastogenesis

    PubMed Central

    Wang, Yongqiang; Kuiper, Johannes W. P.; Glogauer, Michael

    2014-01-01

    Adseverin (Ads), a member of the Gelsolin superfamily of actin binding proteins, regulates the actin cytoskeleton architecture by severing and capping existing filamentous actin (F-actin) strands and nucleating the assembly of new F-actin filaments. Ads has been implicated in cellular secretion, exocytosis and has also been shown to regulate chondrogenesis and megakaryoblastic leukemia cell differentiation. Here we report for the first time that Ads is involved in regulating osteoclastogenesis (OCG). Ads is induced during OCG downstream of RANK-ligand (RANKL) stimulation and is highly expressed in mature osteoclasts. The D5 isoform of Ads is not involved in regulating OCG, as its expression is not induced in response to RANKL. Three clonal Ads knockdown RAW264.7 (RAW) macrophage cell lines with varying degrees of Ads expression and OCG deficiency were generated. The most drastic OCG defect was noted in the clonal cell line with the greatest degree of Ads knockdown as indicated by a lack of TRAcP staining and multinucleation. RNAi mediated knockdown of Ads in osteoclast precursors resulted in distinct morphological changes characterized by altered F-actin distribution and increased filopodia formation. Ads knockdown precursor cells experienced enhanced migration while fusion of knockdown precursors cells was limited. Transient reintroduction of de novo Ads back into the knockdown system was capable of rescuing TRAcP expression but not osteoclast multinucleation most likely due to the transient nature of Ads expression. This preliminary study allows us to conclude that Ads is a RANKL induced early regulator of OCG with a potential role in pre-osteoclast differentiation and fusion. PMID:25275604

  14. Actin Cytoskeleton Contributes to the Elastic Modulus of Embryonic Tendon During Early Development

    PubMed Central

    Schiele, Nathan R.; von Flotow, Friedrich; Tochka, Zachary L.; Hockaday, Laura A.; Marturano, Joseph E.; Thibodeau, Jeffrey J.; Kuo, Catherine K.

    2016-01-01

    Tendon injuries are common and heal poorly. Strategies to regenerate or replace injured tendons are challenged by an incomplete understanding of normal tendon development. Our previous study showed that embryonic tendon elastic modulus increases as a function of developmental stage. Inhibition of enzymatic collagen crosslink formation abrogated increases in tendon elastic modulus at late developmental stages, but did not affect increases in elastic modulus of early stage embryonic tendons. Here, we aimed to identify potential contributors to the mechanical properties of these early stage embryonic tendons. We characterized tendon progenitor cells in early stage embryonic tendons, and the influence of actin cytoskeleton disruption on tissue elastic modulus. Cells were closely packed in embryonic tendons, and did not change in density during early development. We observed an organized network of actin filaments that seemed contiguous between adjacent cells. The actin filaments exhibited a crimp pattern with a period and amplitude that matched the crimp of collagen fibers at each developmental stage. Chemical disruption of the actin cytoskeleton decreased tendon tissue elastic modulus, measured by atomic force microscopy. Our results demonstrate that early developmental stage embryonic tendons possess a well organized actin cytoskeleton network that contributes significantly to tendon tissue mechanical properties. PMID:25721681

  15. F-actin Severing Facilitates Distinct Mechanisms of Stress Relaxation in the Actin Cytoskeleton

    NASA Astrophysics Data System (ADS)

    Kim, Taeyoon; Jung, Wonyeong; Murrell, Michael

    Rheological behaviors of actin cytoskeleton play an important role in physiological processes including cell migration and division. The actin cytoskeleton shows a wide variety of viscoelastic responses to external mechanical cues, such as strain-stiffening and stress relaxation. It has been hypothesized that the stress relaxation originates mainly from transient nature of cross-linkers that connect pairs of F-actins. By contrast, potential impacts of rich F-actin dynamics to the stress relaxation have been neglected in most previous studies. Here, using a computational model, we demonstrated that severing of F-actins induced by buckling during strain-stiffening can facilitate a very distinct mode of stress relaxation in the actin cytoskeleton from that induced by the transient cross-linkers. We also explored conditions where the severing-induced stress relaxation becomes prominent. This finding provides a more complete understanding of rheological behaviors of the actin cytoskeleton. We gratefully acknowledge the support of the National Science Foundation (1434013-CMMI and 1434095-CMMI).

  16. Intra-day up and down of flux density at 4.8 GHz of the quasar S5 1044+71

    NASA Astrophysics Data System (ADS)

    Liu, X.; Lin, M.-Q.; Liu, J.; Cui, L.; Krichbaum, T. P.; Bignall, H.

    2014-04-01

    Following ATEL #5869, we observed the quasar S5 1044+71 at 4.8 GHz in two Intra-day variability (IDV) sessions with the Urumqi 25m radio telescope of Xinjiang Astronomical Observatory (XAO), we find its flux is rising by ~4% on 11-12 Feb. ...

  17. Actin age orchestrates myosin-5 and myosin-6 run lengths.

    PubMed

    Zimmermann, Dennis; Santos, Alicja; Kovar, David R; Rock, Ronald S

    2015-08-01

    Unlike a static and immobile skeleton, the actin cytoskeleton is a highly dynamic network of filamentous actin (F-actin) polymers that continuously turn over. In addition to generating mechanical forces and sensing mechanical deformation, dynamic F-actin networks serve as cellular tracks for myosin motor traffic. However, much of our mechanistic understanding of processive myosins comes from in vitro studies in which motility was studied on pre-assembled and artificially stabilized, static F-actin tracks. In this work, we examine the role of actin dynamics in single-molecule myosin motility using assembling F-actin and two highly processive motors, myosin-5 and myosin-6. These two myosins have distinct functions in the cell and travel in opposite directions along actin filaments [1-3]. Myosin-5 walks toward the barbed ends of F-actin, traveling to sites of actin polymerization at the cell periphery [4]. Myosin-6 walks toward the pointed end of F-actin [5], traveling toward the cell center along older segments of the actin filament. We find that myosin-5 takes 1.3- to 1.5-fold longer runs on ADP•Pi (young) F-actin, whereas myosin-6 takes 1.7- to 3.6-fold longer runs along ADP (old) F-actin. These results suggest that conformational differences between ADP•Pi and ADP F-actin tailor these myosins to walk farther toward their preferred actin filament end. Taken together, these experiments define a new mechanism by which myosin traffic may sort to different F-actin networks depending on filament age. PMID:26190073

  18. The cortical actin determines different susceptibility of naïve and memory CD4+ T cells to HIV-1 cell-to-cell transmission and infection.

    PubMed

    Permanyer, Marc; Pauls, Eduardo; Badia, Roger; Esté, José A; Ballana, Ester

    2013-01-01

    Memory CD4+ T cells are preferentially infected by HIV-1 compared to naïve cells. HIV-1 fusion and entry is a dynamic process in which the cytoskeleton plays an important role by allowing virion internalization and uncoating. Here, we evaluate the role of the cortical actin in cell-to-cell transfer of virus antigens and infection of target CD4+ T cells. Using different actin remodeling compounds we demonstrate that efficiency of HIV-internalization was proportional to the actin polymerization of the target cell. Naïve (CD45RA+) and memory (CD45RA-) CD4+ T cells could be phenotypically differentiated by the degree of cortical actin density and their capacity to capture virus. Thus, the higher cortical actin density of memory CD4+ T cells was associated to increased efficiency of HIV-antigen internalization and the establishment of a productive infection. Conversely, the lower cortical actin density in naïve CD4+ T cells restricted viral antigen transfer and consequently HIV-1 infection. In conclusion, the cortical actin density differentially affects the susceptibility to HIV-1 infection in naïve and memory CD4+ T cells by modulating the efficiency of HIV antigen internalization. PMID:24244453

  19. A complex of ZO-1 and the BAR-domain protein TOCA-1 regulates actin assembly at the tight junction

    PubMed Central

    Van Itallie, Christina M.; Tietgens, Amber Jean; Krystofiak, Evan; Kachar, Bechara; Anderson, James M.

    2015-01-01

    Assembly and sealing of the tight junction barrier are critically dependent on the perijunctional actin cytoskeleton, yet little is known about physical and functional links between barrier-forming proteins and actin. Here we identify a novel functional complex of the junction scaffolding protein ZO-1 and the F-BAR–domain protein TOCA-1. Using MDCK epithelial cells, we show that an alternative splice of TOCA-1 adds a PDZ-binding motif, which binds ZO-1, targeting TOCA-1 to barrier contacts. This isoform of TOCA-1 recruits the actin nucleation–promoting factor N-WASP to tight junctions. CRISPR-Cas9–mediated knockout of TOCA-1 results in increased paracellular flux and delayed recovery in a calcium switch assay. Knockout of TOCA-1 does not alter FRAP kinetics of GFP ZO-1 or occludin, but longer term (12 h) time-lapse microscopy reveals strikingly decreased tight junction membrane contact dynamics in knockout cells compared with controls. Reexpression of TOCA-1 with, but not without, the PDZ-binding motif rescues both altered flux and membrane contact dynamics. Ultrastructural analysis shows actin accumulation at the adherens junction in TOCA-1–knockout cells but unaltered freeze-fracture fibril morphology. Identification of the ZO-1/TOCA-1 complex provides novel insights into the underappreciated dependence of the barrier on the dynamic nature of cell-to-cell contacts and perijunctional actin. PMID:26063734

  20. [Cytoskeletal actin and its associated proteins. Some examples in Protista].

    PubMed

    Guillén, N; Carlier, M F; Brugerolle, G; Tardieux, I; Ausseil, J

    1998-06-01

    Many processes, cell motility being an example, require cells to remodel the actin cytoskeleton in response to both intracellular and extracellular signals. Reorganization of the actin cytoskeleton involves the rapid disassembly and reassembly of actin filaments, a phenomenon regulated by the action of particular actin-binding proteins. In recent years, an interest in studying actin regulation in unicellular organisms has arisen. Parasitic protozoan are among these organisms and studies of the cytoskeleton functions of these protozoan are relevant related to either cell biology or pathogenicity. To discuss recent data in this field, a symposium concerning "Actin and actin-binding proteins in protists" was held on May 8-11 in Paris, France, during the XXXV meeting of the French Society of Protistology. As a brief summary of the symposium we report here findings concerning the in vitro actin dynamic assembly, as well as the characterization of several actin-binding proteins from the parasitic protozoan Entamoeba histolytica, Trichomonas vaginalis and Plasmodium knowlesi. In addition, localization of actin in non-pathogen protists such as Prorocentrum micans and Crypthecodinium cohnii is also presented. The data show that some actin-binding proteins facilitate organization of filaments into higher order structures as pseudopods, while others have regulatory functions, indicating very particular roles for actin-binding proteins. One of the proteins discussed during the symposium, the actin depolymerizing factor ADF, was shown to enhance the treadmilling rate of actin filaments. In vitro, ADF binds to the ADP-bound forms of G-actin and F-actin, thereby participating in and changing the rate of actin assembly. Biochemical approaches allowed the identification of a protein complex formed by HSP/C70-cap32-34 which might also be involved in depolymerization of F-actin in P. knowlesi. Molecular and cellular approaches were used to identify proteins such as ABP-120 and myosin

  1. The structural basis for the intrinsic disorder of the actin filament: the "lateral slipping" model.

    PubMed

    Bremer, A; Millonig, R C; Sütterlin, R; Engel, A; Pollard, T D; Aebi, U

    1991-11-01

    Three-dimensional (3-D) helical reconstructions computed from electron micrographs of negatively stained dispersed F-actin filaments invariably revealed two uninterrupted columns of mass forming the "backbone" of the double-helical filament. The contact between neighboring subunits along the thus defined two long-pitch helical strands was spatially conserved and of high mass density, while the intersubunit contact between them was of lower mass density and varied among reconstructions. In contrast, phalloidinstabilized F-actin filaments displayed higher and spatially more conserved mass density between the two long-pitch helical strands, suggesting that this bicyclic hepta-peptide toxin strengthens the intersubunit contact between the two strands. Consistent with this distinct intersubunit bonding pattern, the two long-pitch helical strands of unstabilized filaments were sometimes observed separated from each other over a distance of two to six subunits, suggesting that the intrastrand intersubunit contact is also physically stronger than the interstrand contact. The resolution of the filament reconstructions, extending to 2.5 nm axially and radially, enabled us to reproducibly "cut out" the F-actin subunit which measured 5.5 nm axially by 6.0 nm tangentially by 3.2 nm radially. The subunit is distinctly polar with a massive "base" pointing towards the "barbed" end of the filament, and a slender "tip" defining its "pointed" end (i.e., relative to the "arrowhead" pattern revealed after stoichiometric decoration of the filaments with myosin subfragment 1). Concavities running approximately parallel to the filament axis both on the inner and outer face of the subunit define a distinct cleft separating the subunit into two domains of similar size: an inner domain confined to radii less than or equal to 2.5-nm forms the uninterrupted backbone of the two long-pitch helical strands, and an outer domain placed at radii of 2-5-nm protrudes radially and thus predominantly

  2. CNS myelin wrapping is driven by actin disassembly.

    PubMed

    Zuchero, J Bradley; Fu, Meng-Meng; Sloan, Steven A; Ibrahim, Adiljan; Olson, Andrew; Zaremba, Anita; Dugas, Jason C; Wienbar, Sophia; Caprariello, Andrew V; Kantor, Christopher; Leonoudakis, Dmitri; Leonoudakus, Dmitri; Lariosa-Willingham, Karen; Kronenberg, Golo; Gertz, Karen; Soderling, Scott H; Miller, Robert H; Barres, Ben A

    2015-07-27

    Myelin is essential in vertebrates for the rapid propagation of action potentials, but the molecular mechanisms driving its formation remain largely unknown. Here we show that the initial stage of process extension and axon ensheathment by oligodendrocytes requires dynamic actin filament assembly by the Arp2/3 complex. Unexpectedly, subsequent myelin wrapping coincides with the upregulation of actin disassembly proteins and rapid disassembly of the oligodendrocyte actin cytoskeleton and does not require Arp2/3. Inducing loss of actin filaments drives oligodendrocyte membrane spreading and myelin wrapping in vivo, and the actin disassembly factor gelsolin is required for normal wrapping. We show that myelin basic protein, a protein essential for CNS myelin wrapping whose role has been unclear, is required for actin disassembly, and its loss phenocopies loss of actin disassembly proteins. Together, these findings provide insight into the molecular mechanism of myelin wrapping and identify it as an actin-independent form of mammalian cell motility. PMID:26166300

  3. The Interaction of Caldesmon with the COOH Terminus of Actin*

    PubMed Central

    Crosbie, Rachelle; Adams, Susan; Chalovich, Joseph M.; Reisler, Emil

    2005-01-01

    Caldesmon interacts with the NH2-terminal region of actin. It is now shown in airfuge centrifugation experiments that modification of the penultimate cysteine residue of actin significantly weakens its binding to caldesmon both in the presence and absence of tropomyosin. Furthermore, as revealed by fluorescence measurements, caldesmon increases the exposure of the COOH-terminal region of actin to the solvent. This effect of caldesmon, like its inhibitory effect on actomyosin ATPase activity, is enhanced in the presence of tropomyosin. Proteolytic removal of the last three COOH-terminal residues of actin, containing the modified cysteine residue, restores the normal binding between caldesmon and actin. These results establish a correlation between the binding of caldesmon to actin and the conformation of the COOH-terminal region of actin and suggest an indirect rather than direct interaction between caldesmon and this part of actin. PMID:1939062

  4. Actin of Beta vulgaris seedlings under the clinorotation

    NASA Astrophysics Data System (ADS)

    Kozeko, L. Ye.

    We study the influence of altered gravity on actin expression in roots of Beta vulguris seedlings grown on the horizontal clinostat (2 rpm) from seed germination for three days. It is shown that the total actin quantity was not influenced. Three actin isoforms are revealed; a relative protein quantity of these isoforms was similar both in clinorotated seedlings and in ones grown in norm. This point to stable expression of actin under the altered gravity conditions.

  5. Dendritic Actin Filament Nucleation Causes Traveling Waves and Patches

    NASA Astrophysics Data System (ADS)

    Carlsson, Anders E.

    2010-06-01

    The polymerization of actin via branching at a cell membrane containing nucleation-promoting factors is simulated using a stochastic-growth methodology. The polymerized-actin distribution displays three types of behavior: (a) traveling waves, (b) moving patches, and (c) random fluctuations. Increasing actin concentration causes a transition from patches to waves. The waves and patches move by a treadmilling mechanism not involving myosin II. The effects of downregulation of key proteins on actin wave behavior are evaluated.

  6. Symmetry breaking in reconstituted actin cortices.

    PubMed

    Abu Shah, Enas; Keren, Kinneret

    2014-01-01

    The actin cortex plays a pivotal role in cell division, in generating and maintaining cell polarity and in motility. In all these contexts, the cortical network has to break symmetry to generate polar cytoskeletal dynamics. Despite extensive research, the mechanisms responsible for regulating cortical dynamics in vivo and inducing symmetry breaking are still unclear. Here we introduce a reconstituted system that self-organizes into dynamic actin cortices at the inner interface of water-in-oil emulsions. This artificial system undergoes spontaneous symmetry breaking, driven by myosin-induced cortical actin flows, which appears remarkably similar to the initial polarization of the embryo in many species. Our in vitro model system recapitulates the rich dynamics of actin cortices in vivo, revealing the basic biophysical and biochemical requirements for cortex formation and symmetry breaking. Moreover, this synthetic system paves the way for further exploration of artificial cells towards the realization of minimal model systems that can move and divide.DOI: http://dx.doi.org/10.7554/eLife.01433.001. PMID:24843007

  7. Curvature and torsion in growing actin networks

    NASA Astrophysics Data System (ADS)

    Shaevitz, Joshua W.; Fletcher, Daniel A.

    2008-06-01

    Intracellular pathogens such as Listeria monocytogenes and Rickettsia rickettsii move within a host cell by polymerizing a comet-tail of actin fibers that ultimately pushes the cell forward. This dense network of cross-linked actin polymers typically exhibits a striking curvature that causes bacteria to move in gently looping paths. Theoretically, tail curvature has been linked to details of motility by considering force and torque balances from a finite number of polymerizing filaments. Here we track beads coated with a prokaryotic activator of actin polymerization in three dimensions to directly quantify the curvature and torsion of bead motility paths. We find that bead paths are more likely to have low rather than high curvature at any given time. Furthermore, path curvature changes very slowly in time, with an autocorrelation decay time of 200 s. Paths with a small radius of curvature, therefore, remain so for an extended period resulting in loops when confined to two dimensions. When allowed to explore a three-dimensional (3D) space, path loops are less evident. Finally, we quantify the torsion in the bead paths and show that beads do not exhibit a significant left- or right-handed bias to their motion in 3D. These results suggest that paths of actin-propelled objects may be attributed to slow changes in curvature, possibly associated with filament debranching, rather than a fixed torque.

  8. Actin-aggregating cucurbitacins from Physocarpus capitatus.

    PubMed

    Maloney, Katherine N; Fujita, Masaki; Eggert, Ulrike S; Schroeder, Frank C; Field, Christine M; Mitchison, Timothy J; Clardy, Jon

    2008-11-01

    Bioassay-guided fractionation of Physocarpus capitatus yielded two new cucurbitacins (3 and 4) along with the known cucurbitacin F (1) and dihydrocucurbitacin F (2). Preliminary mechanism of action studies indicate that the cucurbitacins cause actin aggregates and inhibit cell division. PMID:18959442

  9. Symmetry breaking in reconstituted actin cortices

    PubMed Central

    Abu Shah, Enas; Keren, Kinneret

    2014-01-01

    The actin cortex plays a pivotal role in cell division, in generating and maintaining cell polarity and in motility. In all these contexts, the cortical network has to break symmetry to generate polar cytoskeletal dynamics. Despite extensive research, the mechanisms responsible for regulating cortical dynamics in vivo and inducing symmetry breaking are still unclear. Here we introduce a reconstituted system that self-organizes into dynamic actin cortices at the inner interface of water-in-oil emulsions. This artificial system undergoes spontaneous symmetry breaking, driven by myosin-induced cortical actin flows, which appears remarkably similar to the initial polarization of the embryo in many species. Our in vitro model system recapitulates the rich dynamics of actin cortices in vivo, revealing the basic biophysical and biochemical requirements for cortex formation and symmetry breaking. Moreover, this synthetic system paves the way for further exploration of artificial cells towards the realization of minimal model systems that can move and divide. DOI: http://dx.doi.org/10.7554/eLife.01433.001 PMID:24843007

  10. Competition of two distinct actin networks for actin defines a bistable switch for cell polarization

    PubMed Central

    Lomakin, Alexis J.; Lee, Kun-Chun; Han, Sangyoon J.; Bui, D A.; Davidson, Michael; Mogilner, Alex; Danuser, Gaudenz

    2015-01-01

    Symmetry-breaking polarization enables functional plasticity of cells and tissues and is yet not well understood. Here we show that epithelial cells, hard-wired to maintain a static morphology and to preserve tissue organization, can spontaneously switch to a migratory polarized phenotype upon relaxation of the actomyosin cytoskeleton. We find that myosin-II engages actin in the formation of cortical actomyosin bundles and thus makes it unavailable for deployment in the process of dendritic growth normally driving cell motility. At low contractility regimes epithelial cells polarize in a front-back manner due to emergence of actin retrograde flows powered by dendritic polymerization of actin. Coupled to cell movement, the flows transport myosin-II from the front to the back of the cell, where the motor locally “locks” actin in contractile bundles. This polarization mechanism could be employed by embryonic and cancer epithelial cells in microenvironments where high contractility-driven cell motion is inefficient. PMID:26414403

  11. Non-Straub type actin from molluscan catch muscle.

    PubMed

    Shelud'ko, Nikolay S; Girich, Ulyana V; Lazarev, Stanislav S; Vyatchin, Ilya G

    2016-05-27

    We have developed a method of obtaining natural actin from smooth muscles of the bivalves on the example of the Сrenomytilus grayanus catch muscle. The muscles were previously rigorized to prevent a loss of thin filaments during homogenization and washings. Thin filaments were isolated with a low ionic strength solution in the presence of ATP and sodium pyrophosphate. Surface proteins of thin filaments-tropomyosin, troponin, calponin and some minor actin-binding proteins-were dissociated from actin filaments by increasing the ionic strength to 0.6 M KCL. Natural fibrillar actin obtained in that way depolymerizes easily in low ionic strength solutions commonly used for the extraction of Straub-type actin from acetone powder. Purification of natural actin was carried out by the polymerization-depolymerization cycle. The content of inactivated actin remaining in the supernatant is much less than at a similar purification of Straub-type actin. A comparative investigation was performed between the natural mussel actin and the Straub-type rabbit skeletal actin in terms of the key properties of actin: polymerization, activation of Mg-ATPase activity of myosin, and the electron-microscopic structure of actin polymers. PMID:27120462

  12. Transformation of actin-encapsulating liposomes induced by cytochalasin D.

    PubMed Central

    Miyata, H; Kinosita, K

    1994-01-01

    Liposomes encapsulating actin filaments were prepared by swelling at 0 degrees C lipid film consisting of a mixture of dimyristoyl phosphatidylcholine and cardiolipin (equal amounts by weight) in 100 microM rabbit skeletal muscle actin and 0.5 mM CaCl2 followed by polymerization of actin at 30 degrees C. Liposomes initially assumed either disk or dumbbell shape, but when cytochalasin D was added to the medium surrounding the liposomes, they were found to become spindle shaped. Liposomes containing bovine serum albumin that were given cytochalasin D and actin-containing liposomes that were given dimethylformamide, the solvent for cytochalasin D, did not transform. These results indicated actin-cytochalasin interaction is involved in the transformation process. Falling-ball viscometry and sedimentation analysis of actin solution indicated that cytochalasin cleaved actin filaments and caused depolymerization. The observation of polarized fluorescence of encapsulated actin labeled with acrylodan indicated that the actin filaments in the transformed liposomes aligned along the long axis of the liposomes. Because the actin filaments in the disk- or dumbbell-shaped liposomes formed bundles running along the liposome contour, the transformation was likely to be accompanied by the change in the actin filament arrangement in the liposomes, which was induced by actin-cytochalasin interaction. Images FIGURE 1 FIGURE 2 FIGURE 3 PMID:7948706

  13. Actin cytoskeleton demonstration in Trichomonas vaginalis and in other trichomonads.

    PubMed

    Brugerolle, G; Bricheux, G; Coffe, G

    1996-01-01

    The flagellate form of Trichomonas vaginalis (T v) transforms to amoeboid cells upon adherence to converslips. They grow and their nuclei divide without undergoing cytokinesis, yielding giant cells and a monolayer of T v F-actin was demonstrated in Trichomonas vaginalis by fluorescence microscopy using phalloidin and an anti-actin mAb which labelled the cytoplasm of both the flagellate and amoeboid forms. Comparative electrophoresis and immunoblotting established that the actin band has the same 42 kDa as muscle actin, but 2-D electrophoresis resolved the actin band into four spots; the two major spots observed were superimposable with major muscle actin isoforms. Electron microscopy demonstrated an ectoplasmic microfibrillar layer along the adhesion zone of amoeboid T v adhering to coverslips. Immunogold staining, using anti-actin monoclonal antibodies demonstrated that this layer was mainly composed of actin microfilaments. A comparative immunoblotting study comprising seven trichomonad species showed that all trichomonads studied expressed actin. The mAb Sigma A-4700 specific for an epitope on the actin C-terminal sequence labelled only actin of Trichomonas vaginalis, Tetratrichomonas gallinarum. Trichomitus batrachorum and Hypotrichomonas acosta, but not the actin of Tritrichomonas foetus, Tritrichomonas augusta and Monocercomonas sp. This discrimination between a 'trichomonas branch' and a 'tritrichomonas branch' is congruent with inferred sequence phylogeny from SSu rRNA and with classical phylogeny of trichomonads. PMID:9175265

  14. Tailor-Made Ezrin Actin Binding Domain to Probe Its Interaction with Actin In-Vitro

    PubMed Central

    Shrivastava, Rohini; Köster, Darius; Kalme, Sheetal; Mayor, Satyajit; Neerathilingam, Muniasamy

    2015-01-01

    Ezrin, a member of the ERM (Ezrin/Radixin/Moesin) protein family, is an Actin-plasma membrane linker protein mediating cellular integrity and function. In-vivo study of such interactions is a complex task due to the presence of a large number of endogenous binding partners for both Ezrin and Actin. Further, C-terminal actin binding capacity of the full length Ezrin is naturally shielded by its N-terminal, and only rendered active in the presence of Phosphatidylinositol bisphosphate (PIP2) or phosphorylation at the C-terminal threonine. Here, we demonstrate a strategy for the design, expression and purification of constructs, combining the Ezrin C-terminal actin binding domain, with functional elements such as fusion tags and fluorescence tags to facilitate purification and fluorescence microscopy based studies. For the first time, internal His tag was employed for purification of Ezrin actin binding domain based on in-silico modeling. The functionality (Ezrin-actin interaction) of these constructs was successfully demonstrated by using Total Internal Reflection Fluorescence Microscopy. This design can be extended to other members of the ERM family as well. PMID:25860910

  15. FMNL3 FH2-actin structure gives insight into formin-mediated actin nucleation and elongation

    SciTech Connect

    Thompson, Morgan E; Heimsath, Ernest G; Gauvin, Timothy J; Higgs, Henry N; Kull, F Jon

    2012-12-09

    Formins are actin-assembly factors that act in a variety of actin-based processes. The conserved formin homology 2 (FH2) domain promotes filament nucleation and influences elongation through interaction with the barbed end. FMNL3 is a formin that induces assembly of filopodia but whose FH2 domain is a poor nucleator. The 3.4-Å structure of a mouse FMNL3 FH2 dimer in complex with tetramethylrhodamine-actin uncovers details of formin-regulated actin elongation. We observe distinct FH2 actin-binding regions; interactions in the knob and coiled-coil subdomains are necessary for actin binding, whereas those in the lasso-post interface are important for the stepping mechanism. Biochemical and cellular experiments test the importance of individual residues for function. This structure provides details for FH2-mediated filament elongation by processive capping and supports a model in which C-terminal non-FH2 residues of FMNL3 are required to stabilize the filament nucleus.

  16. VASP is a processive actin polymerase that requires monomeric actin for barbed end association

    PubMed Central

    Hansen, Scott D.

    2010-01-01

    Ena/VASP proteins regulate the actin cytoskeleton during cell migration and morphogenesis and promote assembly of both filopodial and lamellipodial actin networks. To understand the molecular mechanisms underlying their cellular functions we used total internal reflection fluorescence microscopy to visualize VASP tetramers interacting with static and growing actin filaments in vitro. We observed multiple filament binding modes: (1) static side binding, (2) side binding with one-dimensional diffusion, and (3) processive barbed end tracking. Actin monomers antagonize side binding but promote high affinity (Kd = 9 nM) barbed end attachment. In low ionic strength buffers, VASP tetramers are weakly processive (Koff = 0.69 s−1) polymerases that deliver multiple actin monomers per barbed end–binding event and effectively antagonize filament capping. In higher ionic strength buffers, VASP requires profilin for effective polymerase and anti-capping activity. Based on our observations, we propose a mechanism that accounts for all three binding modes and provides a model for how VASP promotes actin filament assembly. PMID:21041447

  17. Neutrophil actin dysfunction is a genetic disorder associated with partial impairment of neutrophil actin assembly in three family members.

    PubMed Central

    Southwick, F S; Dabiri, G A; Stossel, T P

    1988-01-01

    A male infant with a severe neutrophil motility disorder and poorly polymerizable actin in PMN extracts was reported over a decade ago to have neutrophil actin dysfunction (NAD) (1974. N. Engl. J. Med. 291:1093-1099). Polymerized actin (F-actin) content of fixed and permeabilized intact neutrophils from the father, mother, and sister of the NAD index case have been measured using nitrobenzoxadiazole-phallacidin, a fluorescent compound which binds specifically to actin filaments. F-actin content of unstimulated PMN from all three family members was significantly lower than unstimulated control PMN (mean 23.6 +/- 0.4 SEM fluorescent units vs. 32.6 +/- 0.6 for controls). After stimulation with the chemotactic peptide FMLP, maximal F-actin content of NAD family member PMN was below that of controls (52.7 +/- 1.3 vs. 72.6 +/- 1.8). F-actin content of detergent insoluble cytoskeletons after stimulation with FMLP was also significantly lower in PMN from NAD family members as compared with controls (21 +/- 6% vs. 73 +/- 8%). PMN extracts from the father and mother, when treated with 0.6 M KCl, polymerized half as much actin as controls. Whereas diisopropylfluorophosphate treatment of normal PMN decreased actin polymerizability in cell extracts, this treatment increased the assembly of actin in parental PMN extract. Addition of purified actin to NAD extracts failed to reveal an abnormal actin polymerization inhibitory activity, and no obvious structural defect in actin purified from the father's PMNs was noted by HPLC and two dimensional thin layer chromatography of tryptic digests. The present studies of actin assembly in intact PMNs confirm that NAD is associated with a true defect in PMN actin assembly and is a genetic disorder that is recessively inherited. Images PMID:3183050

  18. Crassulacean Acid Metabolism and Photochemical Efficiency of Photosystem II in the Adaxial and Abaxial Parts of the Succulent Leaves of Kalanchoë daigremontiana Grown at Four Photon Flux Densities.

    PubMed

    Winter, K; Awender, G

    1989-07-01

    Kalanchoë daigremontiana, a species possessing crassulacean acid metabolism, was grown at four photon flux densities (1300, 400, 60, and 25 micromole photons per square meter per second). In leaves which had developed at 1300 and 400 micromole photons per square meter per second, CO(2) was mainly incorporated through the lower, shaded leaf surfaces, and the chlorenchyma adjacent to the lower surfaces showed a higher degree of nocturnal acid synthesis than the chlorenchyma adjacent to the upper surfaces. In leaves acclimated to 60 and 25 micromole photons per square meter per second, the gradient in CAM activity was reversed, i.e. more CO(2) was taken up through the upper than through the lower surfaces and nocturnal acidification was higher in the tissue next to the upper surfaces. Total net carbon gain and total nocturnal acid synthesis were highest in leaves which had developed at 400 micromole photons per square meter per second. Chlorophyll content was markedly reduced in leaves which had developed at 1300 micromole photons per square meter per second, especially in the exposed adaxial parts. There was also a sustained reduction in photosystem II photochemical efficiency as indicated by measurements of the ratio of variable over maximum chlorophyll a fluorescence. These findings suggest that, at high growth photon flux densities, the reduced activity of the exposed portions of these succulent leaves is caused by (a) the adverse effects of excess light, (b) together with a genotypic component which favors CO(2) uptake and acid synthesis in the abaxial (lower) leaf parts even when light is not or only marginally excessive. This latter component is predominant at medium photon flux densities, e.g. at 400 micromole photons per square meter per second. It becomes overridden, however, under conditions of deep shade when strongly reduced light levels in the abaxial parts of the leaf chlorenchyma severely limit photosynthesis. PMID:16666903

  19. Structure of a Bud6/Actin Complex Reveals a Novel WH2-like Actin Monomer Recruitment Motif.

    PubMed

    Park, Eunyoung; Graziano, Brian R; Zheng, Wei; Garabedian, Mikael; Goode, Bruce L; Eck, Michael J

    2015-08-01

    In budding yeast, the actin-binding protein Bud6 cooperates with formins Bni1 and Bnr1 to catalyze the assembly of actin filaments. The nucleation-enhancing activity of Bud6 requires both a "core" domain that binds to the formin and a "flank" domain that binds monomeric actin. Here, we describe the structure of the Bud6 flank domain in complex with actin. Two helices in Bud6(flank) interact with actin; one binds in a groove at the barbed end of the actin monomer in a manner closely resembling the helix of WH2 domains, a motif found in many actin nucleation factors. The second helix rises along the face of actin. Mutational analysis verifies the importance of these Bud6-actin contacts for nucleation-enhancing activity. The Bud6 binding site on actin overlaps with that of the formin FH2 domain and is also incompatible with inter-subunit contacts in F-actin, suggesting that Bud6 interacts only transiently with actin monomers during filament nucleation. PMID:26118535

  20. Demonstration of prominent actin filaments in the root columella

    NASA Technical Reports Server (NTRS)

    Collings, D. A.; Zsuppan, G.; Allen, N. S.; Blancaflor, E. B.; Brown, C. S. (Principal Investigator)

    2001-01-01

    The distribution of actin filaments within the gravity-sensing columella cells of plant roots remains poorly understood, with studies over numerous years providing inconsistent descriptions of actin organization in these cells. This uncertainty in actin organization, and thus in actin's role in graviperception and gravisignaling, has led us to investigate actin arrangements in the columella cells of Zea mays L., Medicago truncatula Gaertn., Linum usitatissiilium L. and Nicotianla benthamiana Domin. Actin organization was examined using a combination of optimized immunofluorescence techniques, and an improved fluorochrome-conjugated phalloidin labeling method reliant on 3-maleimidobenzoyl-N-hydroxy-succinimide ester (MBS) cross-linking combined with glycerol permeabilization. Confocal microscopy of root sections labeled with anti-actin antibodies revealed patterns suggestive of actin throughout the columella region. These patterns included short and fragmented actin bundles, fluorescent rings around amyloplasts and intense fluorescence originating from the nucleus. Additionally, confocal microscopy of MBS-stabilized and Alexa Fluor-phalloidin-labeled root sections revealed a previously undetected state of actin organization in the columella. Discrete actin structures surrounded the amyloplasts and prominent actin cables radiated from the nuclear surface toward the cell periphery. Furthermore, the cortex of the columella cells contained fine actin bundles (or single filaments) that had a predominant transverse orientation. We also used confocal microscopy of plant roots expressing endoplasmic reticulum (ER)-targeted green fluorescent protein to demonstrate rapid ER movements within the columella cells, suggesting that the imaged actin network is functional. The successful identification of discrete actin structures in the root columella cells forms the perception and signaling.

  1. Exploring the Stability Limits of Actin and Its Suprastructures

    PubMed Central

    Rosin, Christopher; Erlkamp, Mirko; Ecken, Julian von der; Raunser, Stefan; Winter, Roland

    2014-01-01

    Actin is the main component of the microfilament system in eukaryotic cells and can be found in distinct morphological states. Global (G)-actin is able to assemble into highly organized, supramolecular cellular structures known as filamentous (F)-actin and bundled (B)-actin. To evaluate the structure and stability of G-, F-, and B-actin over a wide range of temperatures and pressures, we used Fourier transform infrared spectroscopy in combination with differential scanning and pressure perturbation calorimetry, small-angle x-ray scattering, laser confocal scanning microscopy, and transmission electron microscopy. Our analysis was designed to provide new (to our knowledge) insights into the stabilizing forces of actin self-assembly and to reveal the stability of the actin polymorphs, including in conditions encountered in extreme environments. In addition, we sought to explain the limited pressure stability of actin self-assembly observed in vivo. G-actin is not only the least temperature-stable but also the least pressure-stable actin species. Under abyssal conditions, where temperatures as low as 1–4°C and pressures up to 1 kbar are reached, G-actin is hardly stable. However, the supramolecular assemblies of actin are stable enough to withstand the extreme conditions usually encountered on Earth. Beyond ∼3–4 kbar, filamentous structures disassemble, and beyond ∼4 kbar, complete dissociation of F-actin structures is observed. Between ∼1 and 2 kbar, some disordering of actin assemblies commences, in agreement with in vivo observations. The limited pressure stability of the monomeric building block seems to be responsible for the suppression of actin assembly in the kbar pressure range. PMID:25517163

  2. Flux Quanta Driven by High-Density Currents in Low-Impurity V3Si and LuNi2B2C: Free Flux Flow and Fluxon-Core Size Effect

    SciTech Connect

    Gapud, Albert A.; Moraes, S.; Khadka, R. P.; Favreau, P.; Henderson, C.; Canfield, P. C.; Kogan, V. G.; Reyes, A. P.; Lumata, L. L.; Christen, David K; Thompson, James R

    2009-01-01

    High-density direct currents are used to drive flux quanta via the Lorentz force toward a highly ordered 'free flux flow' (FFF) dynamic state, made possible by the weak-pinning environment of high-quality, single-crystal samples of two low-T{sub c} superconducting compounds, V{sub 3}Si and LuNi{sub 2}B{sub 2}C. We report the effect of the magnetic field-dependent fluxon-core size on flux flow resistivity {rho}{sub f}. Much progress has been made in minimizing the technical challenges associated with the use of high currents. Attainment of a FFF phase is indicated by the saturation at highest currents of flux flow dissipation levels that are well below the normal-state resistance and have field-dependent values. The field dependence of the corresponding {rho}{sub f} is shown to be consistent with a prediction based on a model for the decrease of fluxon-core size at higher fields in weak-coupling BCS s-wave materials.

  3. Electrostatic interaction map reveals a new binding position for tropomyosin on F-actin.

    PubMed

    Rynkiewicz, Michael J; Schott, Veronika; Orzechowski, Marek; Lehman, William; Fischer, Stefan

    2015-12-01

    Azimuthal movement of tropomyosin around the F-actin thin filament is responsible for muscle activation and relaxation. Recently a model of αα-tropomyosin, derived from molecular-mechanics and electron microscopy of different contractile states, showed that tropomyosin is rather stiff and pre-bent to present one specific face to F-actin during azimuthal transitions. However, a new model based on cryo-EM of troponin- and myosin-free filaments proposes that the interacting-face of tropomyosin can differ significantly from that in the original model. Because resolution was insufficient to assign tropomyosin side-chains, the interacting-face could not be unambiguously determined. Here, we use structural analysis and energy landscapes to further examine the proposed models. The observed bend in seven crystal structures of tropomyosin is much closer in direction and extent to the original model than to the new model. Additionally, we computed the interaction map for repositioning tropomyosin over the F-actin surface, but now extended over a much larger surface than previously (using the original interacting-face). This map shows two energy minima-one corresponding to the "blocked-state" as in the original model, and the other related by a simple 24 Å translation of tropomyosin parallel to the F-actin axis. The tropomyosin-actin complex defined by the second minimum fits perfectly into the recent cryo-EM density, without requiring any change in the interacting-face. Together, these data suggest that movement of tropomyosin between regulatory states does not require interacting-face rotation. Further, they imply that thin filament assembly may involve an interplay between initially seeded tropomyosin molecules growing from distinct binding-site regions on actin. PMID:26286845

  4. Orthogonal (transverse) arrangements of actin in endothelia and fibroblasts

    PubMed Central

    Curtis, Adam; Aitchison, Gregor; Tsapikouni, Theodora

    2006-01-01

    Though actin filaments running across the cell (transverse actin) have been occasionally reported for epithelial cells in groups and for cells growing on fibres, there has been no report heretofore of transverse actin in cells grown on planar substrata. This paper describes evidence in support of this possibility derived from actin staining, polarization microscopy and force measurements. The paper introduces two new methods for detecting the orientation and activity of contractile elements in cells. The orthogonal actin is most obvious in cells grown on groove ridge structures, but can be detected in cells grown on flat surfaces. PMID:17015307

  5. Calcium and actin in the saga of awakening oocytes

    SciTech Connect

    Santella, Luigia Limatola, Nunzia; Chun, Jong T.

    2015-04-24

    The interaction of the spermatozoon with the egg at fertilization remains one of the most fascinating mysteries of life. Much of our scientific knowledge on fertilization comes from studies on sea urchin and starfish, which provide plenty of gametes. Large and transparent, these eggs have served as excellent model systems for studying egg activation and embryo development in seawater, a plain natural medium. Starfish oocytes allow the study of the cortical, cytoplasmic and nuclear changes during the meiotic maturation process, which can also be triggered in vitro by hormonal stimulation. These morphological and biochemical changes ensure successful fertilization of the eggs at the first metaphase. On the other hand, sea urchin eggs are fertilized after the completion of meiosis, and are particularly suitable for the study of sperm–egg interaction, early events of egg activation, and embryonic development, as a large number of mature eggs can be fertilized synchronously. Starfish and sea urchin eggs undergo abrupt changes in the cytoskeleton and ion fluxes in response to the fertilizing spermatozoon. The plasma membrane and cortex of an egg thus represent “excitable media” that quickly respond to the stimulus with the Ca{sup 2+} swings and structural changes. In this article, we review some of the key findings on the rapid dynamic rearrangements of the actin cytoskeleton in the oocyte/egg cortex upon hormonal or sperm stimulation and their roles in the modulation of the Ca{sup 2+} signals and in the control of monospermic fertilization. - Highlights: • Besides microtubules, microfilaments may anchor the nucleus to oocyte surface. • The cortical Ca{sup 2+} flash and wave at fertilization mirror electrical membrane change. • Artificial egg activation lacks microvilli extension in the perivitelline space. • Calcium is necessary but not sufficient for cortical granules exocytosis. • Actin cytoskeleton modulates Ca{sup 2+} release at oocyte maturation

  6. Geometrical and Mechanical Properties Control Actin Filament Organization

    PubMed Central

    Ennomani, Hajer; Théry, Manuel; Nedelec, Francois; Blanchoin, Laurent

    2015-01-01

    The different actin structures governing eukaryotic cell shape and movement are not only determined by the properties of the actin filaments and associated proteins, but also by geometrical constraints. We recently demonstrated that limiting nucleation to specific regions was sufficient to obtain actin networks with different organization. To further investigate how spatially constrained actin nucleation determines the emergent actin organization, we performed detailed simulations of the actin filament system using Cytosim. We first calibrated the steric interaction between filaments, by matching, in simulations and experiments, the bundled actin organization observed with a rectangular bar of nucleating factor. We then studied the overall organization of actin filaments generated by more complex pattern geometries used experimentally. We found that the fraction of parallel versus antiparallel bundles is determined by the mechanical properties of actin filament or bundles and the efficiency of nucleation. Thus nucleation geometry, actin filaments local interactions, bundle rigidity, and nucleation efficiency are the key parameters controlling the emergent actin architecture. We finally simulated more complex nucleation patterns and performed the corresponding experiments to confirm the predictive capabilities of the model. PMID:26016478

  7. Cytoplasmic Actin: Purification and Single Molecule Assembly Assays

    PubMed Central

    Hansen, Scott D.; Zuchero, J. Bradley; Mullins, R. Dyche

    2014-01-01

    The actin cytoskeleton is essential to all eukaryotic cells. In addition to playing important structural roles, assembly of actin into filaments powers diverse cellular processes, including cell motility, cytokinesis, and endocytosis. Actin polymerization is tightly regulated by its numerous cofactors, which control spatial and temporal assembly of actin as well as the physical properties of these filaments. Development of an in vitro model of actin polymerization from purified components has allowed for great advances in determining the effects of these proteins on the actin cytoskeleton. Here we describe how to use the pyrene actin assembly assay to determine the effect of a protein on the kinetics of actin assembly, either directly or as mediated by proteins such as nucleation or capping factors. Secondly, we show how fluorescently labeled phalloidin can be used to visualize the filaments that are created in vitro to give insight into how proteins regulate actin filament structure. Finally, we describe a method for visualizing dynamic assembly and disassembly of single actin filaments and fluorescently labeled actin binding proteins using total internal reflection fluorescence (TIRF) microscopy. PMID:23868587

  8. A complex gene superfamily encodes actin in petunia.

    PubMed Central

    Baird, W V; Meagher, R B

    1987-01-01

    We have shown by several independent criteria that actin is encoded by a very large and complex superfamily of genes in Petunia. Several cDNA and genomic probes encoding actins from diverse organisms (Dictyostelium, Drosophila, chicken and soybean) hybridize to hundreds of restriction fragments in the petunia genome. Actin-hybridizing sequences were isolated from a petunia genomic library at a rate of at least 200 per genome equivalent. Twenty randomly selected actin-hybridizing clones were characterized in more detail. DNA sequence data from four representative and highly divergent clones, PAc2, PAc3, PAc4 and PAc7, demonstrate that these actin-like sequences are related to functional actin genes. Intron positions typical of other known plant actin genes are conserved in these clones. Four of six clones analyzed (PAc1, PAc2, PAc3, PAc4) hybridize to leaf mRNA of the same size (1.7 kb) as that reported for other plant actin mRNAs and to a slightly smaller mRNA species (1.5 kb). Five distinct subfamilies of actin-related genes were characterized which varied in size from a few members to several dozen members. It is clear from our data that other actin gene subfamilies must also exist within the genome. Possible mechanisms of actin gene amplification and genome turnover are discussed. Images Fig. 1. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:3428258

  9. Nuclear F-actin formation and reorganization upon cell spreading.

    PubMed

    Plessner, Matthias; Melak, Michael; Chinchilla, Pilar; Baarlink, Christian; Grosse, Robert

    2015-05-01

    We recently discovered signal-regulated nuclear actin network assembly. However, in contrast to cytoplasmic actin regulation, polymeric nuclear actin structures and functions remain only poorly understood. Here we describe a novel molecular tool to visualize real-time nuclear actin dynamics by targeting the Actin-Chromobody-TagGFP to the nucleus, thus establishing a nuclear Actin-Chromobody. Interestingly, we observe nuclear actin polymerization into dynamic filaments upon cell spreading and fibronectin stimulation, both of which appear to be triggered by integrin signaling. Furthermore, we show that nucleoskeletal proteins such as the LINC (linker of nucleoskeleton and cytoskeleton) complex and components of the nuclear lamina couple cell spreading or integrin activation by fibronectin to nuclear actin polymerization. Spreading-induced nuclear actin polymerization results in serum response factor (SRF)-mediated transcription through nuclear retention of myocardin-related transcription factor A (MRTF-A). Our results reveal a signaling pathway, which links integrin activation by extracellular matrix interaction to nuclear actin polymerization through the LINC complex, and therefore suggest a role for nuclear actin polymerization in the context of cellular adhesion and mechanosensing. PMID:25759381

  10. Distributed actin turnover in the lamellipodium and FRAP kinetics.

    PubMed

    Smith, Matthew B; Kiuchi, Tai; Watanabe, Naoki; Vavylonis, Dimitrios

    2013-01-01

    Studies of actin dynamics at the leading edge of motile cells with single-molecule speckle (SiMS) microscopy have shown a broad distribution of EGFP-actin speckle lifetimes and indicated actin polymerization and depolymerization over an extended region. Other experiments using FRAP with the same EGFP-actin as a probe have suggested, by contrast, that polymerization occurs exclusively at the leading edge. We performed FRAP experiments on XTC cells to compare SiMS to FRAP on the same cell type. We used speckle statistics obtained by SiMS to model the steady-state distribution and kinetics of actin in the lamellipodium. We demonstrate that a model with a single diffuse actin species is in good agreement with FRAP experiments. A model including two species of diffuse actin provides an even better agreement. The second species consists of slowly diffusing oligomers that associate to the F-actin network throughout the lamellipodium or break up into monomers after a characteristic time. Our work motivates studies to test the presence and composition of slowly diffusing actin species that may contribute to local remodeling of the actin network and increase the amount of soluble actin. PMID:23332077

  11. Bundling actin filaments from membranes: some novel players

    PubMed Central

    Thomas, Clément

    2012-01-01

    Progress in live-cell imaging of the cytoskeleton has significantly extended our knowledge about the organization and dynamics of actin filaments near the plasma membrane of plant cells. Noticeably, two populations of filamentous structures can be distinguished. On the one hand, fine actin filaments which exhibit an extremely dynamic behavior basically characterized by fast polymerization and prolific severing events, a process referred to as actin stochastic dynamics. On the other hand, thick actin bundles which are composed of several filaments and which are comparatively more stable although they constantly remodel as well. There is evidence that the actin cytoskeleton plays critical roles in trafficking and signaling at both the cell cortex and organelle periphery but the exact contribution of actin bundles remains unclear. A common view is that actin bundles provide the long-distance tracks used by myosin motors to deliver their cargo to growing regions and accordingly play a particularly important role in cell polarization. However, several studies support that actin bundles are more than simple passive highways and display multiple and dynamic roles in the regulation of many processes, such as cell elongation, polar auxin transport, stomatal and chloroplast movement, and defense against pathogens. The list of identified plant actin-bundling proteins is ever expanding, supporting that plant cells shape structurally and functionally different actin bundles. Here I review the most recently characterized actin-bundling proteins, with a particular focus on those potentially relevant to membrane trafficking and/or signaling. PMID:22936939

  12. Tropomyosin diffusion over actin subunits facilitates thin filament assembly

    PubMed Central

    Fischer, Stefan; Rynkiewicz, Michael J.; Moore, Jeffrey R.; Lehman, William

    2016-01-01

    Coiled-coil tropomyosin binds to consecutive actin-subunits along actin-containing thin filaments. Tropomyosin molecules then polymerize head-to-tail to form cables that wrap helically around the filaments. Little is known about the assembly process that leads to continuous, gap-free tropomyosin cable formation. We propose that tropomyosin molecules diffuse over the actin-filament surface to connect head-to-tail to partners. This possibility is likely because (1) tropomyosin hovers loosely over the actin-filament, thus binding weakly to F-actin and (2) low energy-barriers provide tropomyosin freedom for 1D axial translation on F-actin. We consider that these unique features of the actin-tropomyosin interaction are the basis of tropomyosin cable formation. PMID:26798831

  13. The Structural Basis of Actin Organization by Vinculin and Metavinculin.

    PubMed

    Kim, Laura Y; Thompson, Peter M; Lee, Hyunna T; Pershad, Mihir; Campbell, Sharon L; Alushin, Gregory M

    2016-01-16

    Vinculin is an essential adhesion protein that links membrane-bound integrin and cadherin receptors through their intracellular binding partners to filamentous actin, facilitating mechanotransduction. Here we present an 8.5-Å-resolution cryo-electron microscopy reconstruction and pseudo-atomic model of the vinculin tail (Vt) domain bound to F-actin. Upon actin engagement, the N-terminal "strap" and helix 1 are displaced from the Vt helical bundle to mediate actin bundling. We find that an analogous conformational change also occurs in the H1' helix of the tail domain of metavinculin (MVt) upon actin binding, a muscle-specific splice isoform that suppresses actin bundling by Vt. These data support a model in which metavinculin tunes the actin bundling activity of vinculin in a tissue-specific manner, providing a mechanistic framework for understanding metavinculin mutations associated with hereditary cardiomyopathies. PMID:26493222

  14. The Potential Roles of Actin in The Nucleus

    PubMed Central

    Falahzadeh, Khadijeh; Banaei-Esfahani, Amir; Shahhoseini, Maryam

    2015-01-01

    Over the past few decades, actin’s presence in the nucleus has been demonstrated. Actin is a key protein necessary for different nuclear processes. Although actin is well known for its functional role in dynamic behavior of the cytoskeleton, emerging studies are now highlighting new roles for actin. At the present time there is no doubt about the presence of actin in the nucleus. A number of studies have uncovered the functional involvement of actin in nuclear processes. Actin as one of the nuclear components has its own structured and functional rules, such as nuclear matrix association, chromatin remodeling, transcription by RNA polymerases I, II, III and mRNA processing. In this historical review, we attempt to provide an overview of our current understanding of the functions of actin in the nucleus. PMID:25870830

  15. Regulation of cellular actin architecture by S100A10.

    PubMed

    Jung, M Juliane; Murzik, Ulrike; Wehder, Liane; Hemmerich, Peter; Melle, Christian

    2010-04-15

    Actin structures are involved in several biological processes and the disruption of actin polymerisation induces impaired motility of eukaryotic cells. Different factors are involved in regulation and maintenance of the cytoskeletal actin architecture. Here we show that S100A10 participates in the particular organisation of actin filaments. Down-regulation of S100A10 by specific siRNA triggered a disorganisation of filamentous actin structures without a reduction of the total cellular actin concentration. In contrast, the formation of cytoskeleton structures containing tubulin was unhindered in S100A10 depleted cells. Interestingly, the cellular distribution of annexin A2, an interaction partner of S100A10, was unaffected in S100A10 depleted cells. Cells lacking S100A10 showed an impaired migration activity and were unable to close a scratched wound. Our data provide first insights of S100A10 function as a regulator of the filamentous actin network. PMID:20100475

  16. Wnt Signalling Promotes Actin Dynamics during Axon Remodelling through the Actin-Binding Protein Eps8

    PubMed Central

    Salinas, Patricia C.

    2015-01-01

    Upon arrival at their synaptic targets, axons slow down their growth and extensively remodel before the assembly of presynaptic boutons. Wnt proteins are target-derived secreted factors that promote axonal remodelling and synaptic assembly. In the developing spinal cord, Wnts secreted by motor neurons promote axonal remodelling of NT-3 responsive dorsal root ganglia neurons. Axon remodelling induced by Wnts is characterised by growth cone pausing and enlargement, processes that depend on the re-organisation of microtubules. However, the contribution of the actin cytoskeleton has remained unexplored. Here, we demonstrate that Wnt3a regulates the actin cytoskeleton by rapidly inducing F-actin accumulation in growth cones from rodent DRG neurons through the scaffold protein Dishevelled-1 (Dvl1) and the serine-threonine kinase Gsk3β. Importantly, these changes in actin cytoskeleton occurs before enlargement of the growth cones is evident. Time-lapse imaging shows that Wnt3a increases lamellar protrusion and filopodia velocity. In addition, pharmacological inhibition of actin assembly demonstrates that Wnt3a increases actin dynamics. Through a yeast-two hybrid screen, we identified the actin-binding protein Eps8 as a direct interactor of Dvl1, a scaffold protein crucial for the Wnt signalling pathway. Gain of function of Eps8 mimics Wnt-mediated axon remodelling, whereas Eps8 silencing blocks the axon remodelling activity of Wnt3a. Importantly, blockade of the Dvl1-Eps8 interaction completely abolishes Wnt3a-mediated axonal remodelling. These findings demonstrate a novel role for Wnt-Dvl1 signalling through Eps8 in the regulation of axonal remodeling. PMID:26252776

  17. Single-Molecule Discrimination within Dendritic Spines of Discrete Perisynaptic Sites of Actin Filament Assembly Driving Postsynaptic Reorganization

    NASA Astrophysics Data System (ADS)

    Blanpied, Thomas A.

    2013-03-01

    In the brain, the strength of synaptic transmission between neurons is principally set by the organization of proteins within the receptive, postsynaptic cell. Synaptic strength at an individual site of contact can remain remarkably stable for months or years. However, it also can undergo diverse forms of plasticity which change the strength at that contact independent of changes to neighboring synapses. Such activity-triggered neural plasticity underlies memory storage and cognitive development, and is disrupted in pathological physiology such as addiction and schizophrenia. Much of the short-term regulation of synaptic plasticity occurs within the postsynaptic cell, in small subcompartments surrounding the synaptic contact. Biochemical subcompartmentalization necessary for synapse-specific plasticity is achieved in part by segregation of synapses to micron-sized protrusions from the cell called dendritic spines. Dendritic spines are heavily enriched in the actin cytoskeleton, and regulation of actin polymerization within dendritic spines controls both basal synaptic strength and many forms of synaptic plasticity. However, understanding the mechanism of this control has been difficult because the submicron dimensions of spines limit examination of actin dynamics in the spine interior by conventional confocal microscopy. To overcome this, we developed single-molecule tracking photoactivated localization microscopy (smtPALM) to measure the movement of individual actin molecules within living spines. This revealed inward actin flow from broad areas of the spine plasma membrane, as well as a dense central core of heterogeneous filament orientation. The velocity of single actin molecules along filaments was elevated in discrete regions within the spine, notably near the postsynaptic density but surprisingly not at the endocytic zone which is involved in some forms of plasticity. We conclude that actin polymerization is initiated at many well-separated foci within

  18. Cytoarchitecture of Kirsten sarcoma virus-transformed rat kidney fibroblasts: butyrate-induced reorganization within the actin microfilament network.

    PubMed

    Ryan, M P; Higgins, P J

    1988-10-01

    Murine sarcoma virus-transformed rat fibroblasts (KNRK cells) undergo marked cytoarchitectural reorganization during in vitro exposure to sodium-n-butyrate (NaB) resulting in restoration of (1) a more typical fibroblastoid morphology, (2) proper cell-to-cell orientation, and (3) substratum adherence. Augmented cell spreading, involving greater than 90% of the population, was a function of culture density and time of exposure to NaB (2 mM final concentration). Induced cell spreading reflected a 2.5- to 3.0-fold increase in both total cellular actin content and deposition of actin into the detergent-resistant cytoskeleton. Cytoskeletal actin deposition in response to NaB was accompanied by the formation of occasionally dense, parallel alignments of F-actin-containing microfilaments and by a dramatic increase in the size and incidence of actin-enriched membrane ruffles. Long-term NaB-treated cells exhibited parallel orientations of microfilaments similar to those found in untransformed fibroblasts. Increased cytoskeletal actin occurred within 24 hr of NaB exposure, correlating with the initial reorganization of actin-containing microfilaments detected microscopically, and reflected concomitant 3-fold increases in cellular alpha-actinin and fibronectin content. In contrast, the amount of vimentin, tropomyosin, and tubulin in NaB-treated cells was significantly decreased. NaB-induced morphologic restructuring of sarcoma virus-transformed fibroblasts, thus, impacts on all three basic cytoskeletal systems. Selective increases, however, were evident in particular cytoskeletal proteins (actin, alpha-actinin, fibronectin) implicated in microfilament networking and cell spreading. PMID:2844835

  19. Distinct Functional Interactions between Actin Isoforms and Nonsarcomeric Myosins

    PubMed Central

    Müller, Mirco; Diensthuber, Ralph P.; Chizhov, Igor; Claus, Peter; Heissler, Sarah M.; Preller, Matthias; Taft, Manuel H.; Manstein, Dietmar J.

    2013-01-01

    Despite their near sequence identity, actin isoforms cannot completely replace each other in vivo and show marked differences in their tissue-specific and subcellular localization. Little is known about isoform-specific differences in their interactions with myosin motors and other actin-binding proteins. Mammalian cytoplasmic β- and γ-actin interact with nonsarcomeric conventional myosins such as the members of the nonmuscle myosin-2 family and myosin-7A. These interactions support a wide range of cellular processes including cytokinesis, maintenance of cell polarity, cell adhesion, migration, and mechano-electrical transduction. To elucidate differences in the ability of isoactins to bind and stimulate the enzymatic activity of individual myosin isoforms, we characterized the interactions of human skeletal muscle α-actin, cytoplasmic β-actin, and cytoplasmic γ-actin with human myosin-7A and nonmuscle myosins-2A, -2B and -2C1. In the case of nonmuscle myosins-2A and -2B, the interaction with either cytoplasmic actin isoform results in 4-fold greater stimulation of myosin ATPase activity than was observed in the presence of α-skeletal muscle actin. Nonmuscle myosin-2C1 is most potently activated by β-actin and myosin-7A by γ-actin. Our results indicate that β- and γ-actin isoforms contribute to the modulation of nonmuscle myosin-2 and myosin-7A activity and thereby to the spatial and temporal regulation of cytoskeletal dynamics. FRET-based analyses show efficient copolymerization abilities for the actin isoforms in vitro. Experiments with hybrid actin filaments show that the extent of actomyosin coupling efficiency can be regulated by the isoform composition of actin filaments. PMID:23923011

  20. Isolation and characterization of six different chicken actin genes.

    PubMed Central

    Chang, K S; Zimmer, W E; Bergsma, D J; Dodgson, J B; Schwartz, R J

    1984-01-01

    Genes representing six different actin isoforms were isolated from a chicken genomic library. Cloned actin cDNAs as well as tissue-specific mRNAs enriched in different actin species were used as hybridization probes to group individual actin genomic clones by their relative thermal stability. Restriction maps showed that these actin genes were derived from separate and nonoverlapping regions of genomic DNA. Of the six isolated genes, five included sequences from both the 5' and 3' ends of the actin-coding area. Amino acid sequence analysis from both the NH2- and COOH-terminal regions provided for the unequivocal identification of these genes. The striated isoforms were represented by the isolated alpha-skeletal, alpha-cardiac, and alpha-smooth muscle actin genes. The nonmuscle isoforms included the beta-cytoplasmic actin gene and an actin gene fragment which lacked the 5' coding and flanking sequence; presumably, this region of DNA was removed from this gene during construction of the genomic library. Unexpectedly, a third nonmuscle chicken actin gene was found which resembled the amphibian type 5 actin isoform (J. Vandekerckhove, W. W. Franke, and K. Weber, J. Mol. Biol., 152:413-426). This nonmuscle actin type has not been previously detected in warm-blooded vertebrates. We showed that interspersed, repeated DNA sequences closely flanked the alpha-skeletal, alpha-cardiac, beta-, and type 5-like actin genes. The repeated DNA sequences which surround the alpha-skeletal actin-coding regions were not related to repetitious DNA located on the other actin genes. Analysis of genomic DNA blots showed that the chicken actin multigene family was represented by 8 to 10 separate coding loci. The six isolated actin genes corresponded to 7 of 11 genomic EcoRI fragments. Only the alpha-smooth muscle actin gene was shown to be split by an EcoRI site. Thus, in the chicken genome each actin isoform appeared to be encoded by a single gene. Images PMID:6513927

  1. Actinic keratoses: past, present and future.

    PubMed

    Fenske, Neil Alan; Spencer, James; Adam, Friedman

    2010-05-01

    Actinic keratoses (AKs) are cutaneous neoplasms composed of proliferations of cytologically aberrant, epidermal keratinocytes caused by prolonged exposure to ultraviolet radiation. Combining the evidence that AKs are the second most common reason for visits to the dermatologist and it is generally believed that they should be treated, it is no surprise that the direct cost of the management of actinic keratoses in the United States (U.S.) is exceedingly high. There are currently numerous treatment modalities with more on the way as there is a demand for formulating newer, cheaper, less painful and less invasive means. The future of AK treatment involves both the continued investigation of current novel therapies, as well as the development of new treatment modalities. PMID:20518359

  2. Actin-mediated motion of meiotic chromosomes

    PubMed Central

    Koszul, R.; Kim, K. P.; Prentiss, M.; Kleckner, N.; Kameoka, S.

    2008-01-01

    Summary Chromosome movement is prominent during meiosis. Here, using a combination of in vitro and in vivo approaches, we elucidate the basis for dynamic mid-prophase chromosome movement in budding yeast. Diverse finding reveal a process in which, at the pachytene stage, individual telomere/nuclear envelope (NE) ensembles attach passively to, and then move in concert with, nucleus-hugging actin cables that are continuous with the global cytoskeletal actin network. Other chromosomes move in concert with lead chromosome(s). The same process, in modulated form, explains the zygotene "bouquet" configuration in which, immediately preceding pachytene, chromosome ends colocalize dynamically in a restricted region of the NE. Mechanical properties of the system and biological roles of mid-prophase movement for meiosis, including recombination, are discussed. PMID:18585353

  3. Regulation of actin nucleation and autophagosome formation.

    PubMed

    Coutts, Amanda S; La Thangue, Nicholas B

    2016-09-01

    Autophagy is a process of self-eating, whereby cytosolic constituents are enclosed by a double-membrane vesicle before delivery to the lysosome for degradation. This is an important process which allows for recycling of nutrients and cellular components and thus plays a critical role in normal cellular homeostasis as well as cell survival during stresses such as starvation or hypoxia. A large number of proteins regulate various stages of autophagy in a complex and still incompletely understood series of events. In this review, we will discuss recent studies which provide a growing body of evidence that actin dynamics and proteins that influence actin nucleation play an important role in the regulation of autophagosome formation and maturation. PMID:27147468

  4. Caffeine relaxes smooth muscle through actin depolymerization.

    PubMed

    Tazzeo, Tracy; Bates, Genevieve; Roman, Horia Nicolae; Lauzon, Anne-Marie; Khasnis, Mukta D; Eto, Masumi; Janssen, Luke J

    2012-08-15

    Caffeine is sometimes used in cell physiological studies to release internally stored Ca(2+). We obtained evidence that caffeine may also act through a different mechanism that has not been previously described and sought to examine this in greater detail. We ruled out a role for phosphodiesterase (PDE) inhibition, since the effect was 1) not reversed by inhibiting PKA or adenylate cyclase; 2) not exacerbated by inhibiting PDE4; and 3) not mimicked by submillimolar caffeine nor theophylline, both of which are sufficient to inhibit PDE. Although caffeine is an agonist of bitter taste receptors, which in turn mediate bronchodilation, its relaxant effect was not mimicked by quinine. After permeabilizing the membrane using β-escin and depleting the internal Ca(2+) store using A23187, we found that 10 mM caffeine reversed tone evoked by direct application of Ca(2+), suggesting it functionally antagonizes the contractile apparatus. Using a variety of molecular techniques, we found that caffeine did not affect phosphorylation of myosin light chain (MLC) by MLC kinase, actin-filament motility catalyzed by MLC kinase, phosphorylation of CPI-17 by either protein kinase C or RhoA kinase, nor the activity of MLC-phosphatase. However, we did obtain evidence that caffeine decreased actin filament binding to phosphorylated myosin heads and increased the ratio of globular to filamentous actin in precontracted tissues. We conclude that, in addition to its other non-RyR targets, caffeine also interferes with actin function (decreased binding by myosin, possibly with depolymerization), an effect that should be borne in mind in studies using caffeine to probe excitation-contraction coupling in smooth muscle. PMID:22683573

  5. Arabidopsis CROLIN1, a Novel Plant Actin-binding Protein, Functions in Cross-linking and Stabilizing Actin Filaments*

    PubMed Central

    Jia, Honglei; Li, Jisheng; Zhu, Jingen; Fan, Tingting; Qian, Dong; Zhou, Yuelong; Wang, Jiaojiao; Ren, Haiyun; Xiang, Yun; An, Lizhe

    2013-01-01

    Higher order actin filament structures are necessary for cytoplasmic streaming, organelle movement, and other physiological processes. However, the mechanism by which the higher order cytoskeleton is formed in plants remains unknown. In this study, we identified a novel actin-cross-linking protein family (named CROLIN) that is well conserved only in the plant kingdom. There are six isovariants of CROLIN in the Arabidopsis genome, with CROLIN1 specifically expressed in pollen. In vitro biochemical analyses showed that CROLIN1 is a novel actin-cross-linking protein with binding and stabilizing activities. Remarkably, CROLIN1 can cross-link actin bundles into actin networks. CROLIN1 loss of function induces pollen germination and pollen tube growth hypersensitive to latrunculin B. All of these results demonstrate that CROLIN1 may play an important role in stabilizing and remodeling actin filaments by binding to and cross-linking actin filaments. PMID:24072702

  6. Lamellipodin promotes actin assembly by clustering Ena/VASP proteins and tethering them to actin filaments

    PubMed Central

    Hansen, Scott D; Mullins, R Dyche

    2015-01-01

    Enabled/Vasodilator (Ena/VASP) proteins promote actin filament assembly at multiple locations, including: leading edge membranes, focal adhesions, and the surface of intracellular pathogens. One important Ena/VASP regulator is the mig-10/Lamellipodin/RIAM family of adaptors that promote lamellipod formation in fibroblasts and drive neurite outgrowth and axon guidance in neurons. To better understand how MRL proteins promote actin network formation we studied the interactions between Lamellipodin (Lpd), actin, and VASP, both in vivo and in vitro. We find that Lpd binds directly to actin filaments and that this interaction regulates its subcellular localization and enhances its effect on VASP polymerase activity. We propose that Lpd delivers Ena/VASP proteins to growing barbed ends and increases their polymerase activity by tethering them to filaments. This interaction represents one more pathway by which growing actin filaments produce positive feedback to control localization and activity of proteins that regulate their assembly. DOI: http://dx.doi.org/10.7554/eLife.06585.001 PMID:26295568

  7. Characterization of actin filament deformation in response to actively driven microspheres propagated through entangled actin networks

    NASA Astrophysics Data System (ADS)

    Falzone, Tobias; Blair, Savanna; Robertson-Anderson, Rae

    2014-03-01

    The semi-flexible biopolymer actin is a ubiquitous component of nearly all biological organisms, playing an important role in many biological processes such as cell structure and motility, cancer invasion and metastasis, muscle contraction, and cell signaling. Concentrated actin networks possess unique viscoelastic properties that have been the subject of much theoretical and experimental work. However, much is still unknown regarding the correlation of the applied stress on the network to the induced filament strain at the molecular level. Here, we use dual optical traps alongside fluorescence microscopy to carry out active microrheology measurements that link mechanical stress to structural response at the micron scale. Specifically, we actively drive microspheres through entangled actin networks while simultaneously measuring the force the surrounding filaments exert on the sphere and visualizing the deformation and subsequent relaxation of fluorescent labeled filaments within the network. These measurements, which provide much needed insight into the link between stress and strain in actin networks, are critical for clarifying our theoretical understanding of the complex viscoelastic behavior exhibited in actin networks.

  8. Virulent Burkholderia species mimic host actin polymerases to drive actin-based motility

    PubMed Central

    Benanti, Erin L.; Nguyen, Catherine M.; Welch, Matthew D.

    2015-01-01

    Summary Burkholderia pseudomallei and B. mallei are bacterial pathogens that cause melioidosis and glanders, while their close relative B. thailandensis is nonpathogenic. All use the trimeric autotransporter BimA to facilitate actin-based motility, host cell fusion and dissemination. Here, we show that BimA orthologs mimic different host actin-polymerizing proteins. B. thailandensis BimA activates the host Arp2/3 complex. In contrast, B. pseudomallei and B. mallei BimA mimic host Ena/VASP actin polymerases in their ability to nucleate, elongate and bundle filaments by associating with barbed ends, as well as in their use of WH2 motifs and oligomerization for activity. Mechanistic differences among BimA orthologs resulted in distinct actin filament organization and motility parameters, which affected the efficiency of cell fusion during infection. Our results identify bacterial Ena/VASP mimics and reveal that pathogens imitate the full spectrum of host actin-polymerizing pathways, suggesting that mimicry of different polymerization mechanisms influences key parameters of infection. PMID:25860613

  9. The ROSAT-ESO Flux-Limited X-ray (REFLEX) galaxy cluster survey - VI. Constraints on the cosmic matter density from the KL power spectrum

    NASA Astrophysics Data System (ADS)

    Schuecker, Peter; Guzzo, Luigi; Collins, Chris A.; Böhringer, Hans

    2002-09-01

    The Karhunen-Loéve (KL) eigenvectors and eigenvalues of the sample correlation matrix are used to analyse the spatial fluctuations of the REFLEX clusters of galaxies. The method avoids the disturbing effects of correlated power spectral densities that affect all previous cluster measurements on Gpc scales. Comprehensive tests use a large set of independent REFLEX-like mock cluster samples extracted from the Hubble Volume Simulation. It is found that unbiased measurements on Gpc scales are possible with the REFLEX data. The distribution of the KL eigenvalues is consistent with a Gaussian random field on the 93.4 per cent confidence level. Assuming spatially flat cold dark matter models, the marginalization of the likelihood contours over different sample volumes, fiducial cosmologies, mass-X-ray luminosity relations and baryon densities, yields a 95.4 per cent confidence interval for the matter density of 0.03 < Ωmh2 < 0.19. The N-body simulations show that cosmic variance, although difficult to estimate, is expected to increase the confidence intervals by about 50 per cent.

  10. Computational Study of the Binding Mechanism of Actin-Depolymerizing Factor 1 with Actin in Arabidopsis thaliana

    PubMed Central

    Wang, Xue; Dong, Chun-Hai; Yang, Jian Ming; Yao, Xiao Jun

    2016-01-01

    Actin is a highly conserved protein. It plays important roles in cellular function and exists either in the monomeric (G-actin) or polymeric form (F-actin). Members of the actin-depolymerizing factor (ADF)/cofilin protein family bind to both G-actin and F-actin and play vital roles in actin dynamics by manipulating the rates of filament polymerization and depolymerization. It has been reported that the S6D and R98A/K100A mutants of actin-depolymerizing factor 1 (ADF1) in Arabidopsis thaliana decreased the binding affinity of ADF for the actin monomer. To investigate the binding mechanism and dynamic behavior of the ADF1–actin complex, we constructed a homology model of the AtADF1–actin complex based on the crystal structure of AtADF1 and the twinfilin C-terminal ADF-H domain in a complex with a mouse actin monomer. The model was then refined for subsequent molecular dynamics simulations. Increased binding energy of the mutated system was observed using the Molecular Mechanics Generalized Born Surface Area and Poisson–Boltzmann Surface Area (MM-GB/PBSA) methods. To determine the residues that make decisive contributions to the ADF1 actin-binding affinity, per-residue decomposition and computational alanine scanning analyses were performed, which provided more detailed information on the binding mechanism. Root-mean-square fluctuation and principal component analyses confirmed that the S6D and R98A/K100A mutants induced an increased conformational flexibility. The comprehensive molecular insight gained from this study is of great importance for understanding the binding mechanism of ADF1 and G-actin. PMID:27414648

  11. Computational Study of the Binding Mechanism of Actin-Depolymerizing Factor 1 with Actin in Arabidopsis thaliana.

    PubMed

    Du, Juan; Wang, Xue; Dong, Chun-Hai; Yang, Jian Ming; Yao, Xiao Jun

    2016-01-01

    Actin is a highly conserved protein. It plays important roles in cellular function and exists either in the monomeric (G-actin) or polymeric form (F-actin). Members of the actin-depolymerizing factor (ADF)/cofilin protein family bind to both G-actin and F-actin and play vital roles in actin dynamics by manipulating the rates of filament polymerization and depolymerization. It has been reported that the S6D and R98A/K100A mutants of actin-depolymerizing factor 1 (ADF1) in Arabidopsis thaliana decreased the binding affinity of ADF for the actin monomer. To investigate the binding mechanism and dynamic behavior of the ADF1-actin complex, we constructed a homology model of the AtADF1-actin complex based on the crystal structure of AtADF1 and the twinfilin C-terminal ADF-H domain in a complex with a mouse actin monomer. The model was then refined for subsequent molecular dynamics simulations. Increased binding energy of the mutated system was observed using the Molecular Mechanics Generalized Born Surface Area and Poisson-Boltzmann Surface Area (MM-GB/PBSA) methods. To determine the residues that make decisive contributions to the ADF1 actin-binding affinity, per-residue decomposition and computational alanine scanning analyses were performed, which provided more detailed information on the binding mechanism. Root-mean-square fluctuation and principal component analyses confirmed that the S6D and R98A/K100A mutants induced an increased conformational flexibility. The comprehensive molecular insight gained from this study is of great importance for understanding the binding mechanism of ADF1 and G-actin. PMID:27414648

  12. Gelsolin mediates calcium-dependent disassembly of Listeria actin tails

    PubMed Central

    Larson, Laura; Arnaudeau, Serge; Gibson, Bruce; Li, Wei; Krause, Ryoko; Hao, Binghua; Bamburg, James R.; Lew, Daniel P.; Demaurex, Nicolas; Southwick, Frederick

    2005-01-01

    The role of intracellular Ca2+ in the regulation of actin filament assembly and disassembly has not been clearly defined. We show that reduction of intracellular free Ca2+ concentration ([Ca2+]i) to <40 nM in Listeria monocytogenes-infected, EGFP–actin-transfected Madin–Darby canine kidney cells results in a 3-fold lengthening of actin filament tails. This increase in tail length is the consequence of marked slowing of the actin filament disassembly rate, without a significant change in assembly rate. The Ca2+-sensitive actin-severing protein gelsolin concentrates in the Listeria rocket tails at normal resting [Ca2+]i and disassociates from the tails when [Ca2+]i is lowered. Reduction in [Ca2+]i also blocks the severing activity of gelsolin, but not actin-depolymerizing factor (ADF)/cofilin microinjected into Listeria-infected cells. In Xenopus extracts, Listeria tail lengths are also calcium-sensitive, markedly shortening on addition of calcium. Immunodepletion of gelsolin, but not Xenopus ADF/cofilin, eliminates calcium-sensitive actin-filament shortening. Listeria tail length is also calcium-insensitive in gelsolin-null mouse embryo fibroblasts. We conclude that gelsolin is the primary Ca2+-sensitive actin filament recycling protein in the cell and is capable of enhancing Listeria actin tail disassembly at normal resting [Ca2+]i (145 nM). These experiments illustrate the unique and complementary functions of gelsolin and ADF/cofilin in the recycling of actin filaments. PMID:15671163

  13. The neuronal and actin commitment: Why do neurons need rings?

    PubMed

    Leite, Sérgio Carvalho; Sousa, Mónica Mendes

    2016-09-01

    The role of the actin cytoskeleton in neurons has been extensively studied in actin-enriched compartments such as the growth cone and dendritic spines. The recent discovery of actin rings in the axon shaft and in dendrites, together with the identification of axon actin trails, has advanced our understanding on actin organization and dynamics in neurons. However, specifically in the case of actin rings, the mechanisms regulating their nucleation and assembly, and the functions that they may exert in axons and dendrites remain largely unexplored. Here we discuss the possible structural, mechanistic and functional properties of the subcortical neuronal cytoskeleton putting the current knowledge in perspective with the information available on actin rings formed in other biological contexts, and with the organization of actin-spectrin lattices in other cell types. The detailed analysis of these novel neuronal actin ring structures, together with the elucidation of the function of actin-binding proteins in neuron biology, has a large potential to uncover new mechanisms of neuronal function under normal conditions that may have impact in our understanding of axon degeneration and regeneration. © 2016 Wiley Periodicals, Inc. PMID:26784007

  14. Excitable actin dynamics in lamellipodial protrusion and retraction.

    PubMed

    Ryan, Gillian L; Petroccia, Heather M; Watanabe, Naoki; Vavylonis, Dimitrios

    2012-04-01

    Many animal cells initiate crawling by protruding lamellipodia, consisting of a dense network of actin filaments, at their leading edge. We imaged XTC cells that exhibit flat lamellipodia on poly-L-lysine-coated coverslips. Using active contours, we tracked the leading edge and measured the total amount of F-actin by summing the pixel intensities within a 5-μm band. We observed protrusion and retraction with period 130-200 s and local wavelike features. Positive (negative) velocities correlated with minimum (maximum) integrated actin concentration. Approximately constant retrograde flow indicated that protrusions and retractions were driven by fluctuations of the actin polymerization rate. We present a model of these actin dynamics as an excitable system in which a diffusive, autocatalytic activator causes actin polymerization; F-actin accumulation in turn inhibits further activator accumulation. Simulations of the model reproduced the pattern of actin polymerization seen in experiments. To explore the model's assumption of an autocatalytic activation mechanism, we imaged cells expressing markers for both F-actin and the p21 subunit of the Arp2/3 complex. We found that integrated Arp2/3-complex concentrations spike several seconds before spikes of F-actin concentration. This suggests that the Arp2/3 complex participates in an activation mechanism that includes additional diffuse components. Response of cells to stimulation by fetal calf serum could be reproduced by the model, further supporting the proposed dynamical picture. PMID:22500749

  15. Sequence and comparative genomic analysis of actin-related proteins.

    PubMed

    Muller, Jean; Oma, Yukako; Vallar, Laurent; Friederich, Evelyne; Poch, Olivier; Winsor, Barbara

    2005-12-01

    Actin-related proteins (ARPs) are key players in cytoskeleton activities and nuclear functions. Two complexes, ARP2/3 and ARP1/11, also known as dynactin, are implicated in actin dynamics and in microtubule-based trafficking, respectively. ARP4 to ARP9 are components of many chromatin-modulating complexes. Conventional actins and ARPs codefine a large family of homologous proteins, the actin superfamily, with a tertiary structure known as the actin fold. Because ARPs and actin share high sequence conservation, clear family definition requires distinct features to easily and systematically identify each subfamily. In this study we performed an in depth sequence and comparative genomic analysis of ARP subfamilies. A high-quality multiple alignment of approximately 700 complete protein sequences homologous to actin, including 148 ARP sequences, allowed us to extend the ARP classification to new organisms. Sequence alignments revealed conserved residues, motifs, and inserted sequence signatures to define each ARP subfamily. These discriminative characteristics allowed us to develop ARPAnno (http://bips.u-strasbg.fr/ARPAnno), a new web server dedicated to the annotation of ARP sequences. Analyses of sequence conservation among actins and ARPs highlight part of the actin fold and suggest interactions between ARPs and actin-binding proteins. Finally, analysis of ARP distribution across eukaryotic phyla emphasizes the central importance of nuclear ARPs, particularly the multifunctional ARP4. PMID:16195354

  16. Actin is required for IFT regulation in Chlamydomonas reinhardtii

    PubMed Central

    Avasthi, Prachee; Onishi, Masayuki; Karpiak, Joel; Yamamoto, Ryosuke; Mackinder, Luke; Jonikas, Martin C.; Sale, Winfield S.; Shoichet, Brian; Pringle, John R.; Marshall, Wallace F.

    2014-01-01

    Summary Assembly of cilia and flagella requires intraflagellar transport (IFT), a highly regulated kinesin-based transport system that moves cargo from the basal body to the tip of flagella [1]. The recruitment of IFT components to basal bodies is a function of flagellar length, with increased recruitment in rapidly growing short flagella [2]. The molecular pathways regulating IFT are largely a mystery. Since actin network disruption leads to changes in ciliary length and number, actin has been proposed to have a role in ciliary assembly. However, the mechanisms involved are unknown. In Chlamydomonas reinhardtii, conventional actin is found in both the cell body and the inner dynein arm complexes within flagella [3, 4]. Previous work showed that treating Chlamydomonas cells with the actin-depolymerizing compound cytochalasin D resulted in reversible flagellar shortening [5], but how actin is related to flagellar length or assembly remains unknown. Here, we utilize small-molecule inhibitors and genetic mutants to analyze the role of actin dynamics in flagellar assembly in Chlamydomonas reinhardtii. We demonstrate that actin plays a role in IFT recruitment to basal bodies during flagellar elongation, and that when actin is perturbed, the normal dependence of IFT recruitment on flagellar length is lost. We also find that actin is required for sufficient entry of IFT material into flagella during assembly. These same effects are recapitulated with a myosin inhibitor suggesting actin may act via myosin in a pathway by which flagellar assembly is regulated by flagellar length. PMID:25155506

  17. Actin is required for IFT regulation in Chlamydomonas reinhardtii.

    PubMed

    Avasthi, Prachee; Onishi, Masayuki; Karpiak, Joel; Yamamoto, Ryosuke; Mackinder, Luke; Jonikas, Martin C; Sale, Winfield S; Shoichet, Brian; Pringle, John R; Marshall, Wallace F

    2014-09-01

    Assembly of cilia and flagella requires intraflagellar transport (IFT), a highly regulated kinesin-based transport system that moves cargo from the basal body to the tip of flagella [1]. The recruitment of IFT components to basal bodies is a function of flagellar length, with increased recruitment in rapidly growing short flagella [2]. The molecular pathways regulating IFT are largely a mystery. Because actin network disruption leads to changes in ciliary length and number, actin has been proposed to have a role in ciliary assembly. However, the mechanisms involved are unknown. In Chlamydomonas reinhardtii, conventional actin is found in both the cell body and the inner dynein arm complexes within flagella [3, 4]. Previous work showed that treating Chlamydomonas cells with the actin-depolymerizing compound cytochalasin D resulted in reversible flagellar shortening [5], but how actin is related to flagellar length or assembly remains unknown. Here we utilize small-molecule inhibitors and genetic mutants to analyze the role of actin dynamics in flagellar assembly in Chlamydomonas reinhardtii. We demonstrate that actin plays a role in IFT recruitment to basal bodies during flagellar elongation and that when actin is perturbed, the normal dependence of IFT recruitment on flagellar length is lost. We also find that actin is required for sufficient entry of IFT material into flagella during assembly. These same effects are recapitulated with a myosin inhibitor, suggesting that actin may act via myosin in a pathway by which flagellar assembly is regulated by flagellar length. PMID:25155506

  18. Tropomyosin - master regulator of actin filament function in the cytoskeleton.

    PubMed

    Gunning, Peter W; Hardeman, Edna C; Lappalainen, Pekka; Mulvihill, Daniel P

    2015-08-15

    Tropomyosin (Tpm) isoforms are the master regulators of the functions of individual actin filaments in fungi and metazoans. Tpms are coiled-coil parallel dimers that form a head-to-tail polymer along the length of actin filaments. Yeast only has two Tpm isoforms, whereas mammals have over 40. Each cytoskeletal actin filament contains a homopolymer of Tpm homodimers, resulting in a filament of uniform Tpm composition along its length. Evidence for this 'master regulator' role is based on four core sets of observation. First, spatially and functionally distinct actin filaments contain different Tpm isoforms, and recent data suggest that members of the formin family of actin filament nucleators can specify which Tpm isoform is added to the growing actin filament. Second, Tpms regulate whole-organism physiology in terms of morphogenesis, cell proliferation, vesicle trafficking, biomechanics, glucose metabolism and organ size in an isoform-specific manner. Third, Tpms achieve these functional outputs by regulating the interaction of actin filaments with myosin motors and actin-binding proteins in an isoform-specific manner. Last, the assembly of complex structures, such as stress fibers and podosomes involves the collaboration of multiple types of actin filament specified by their Tpm composition. This allows the cell to specify actin filament function in time and space by simply specifying their Tpm isoform composition. PMID:26240174

  19. Actin Turnover-Mediated Gravity Response in Maize Root Apices

    PubMed Central

    Mancuso, Stefano; Barlow, Peter W; Volkmann, Dieter

    2006-01-01

    The dynamic actin cytoskeleton has been proposed to be linked to gravity sensing in plants but the mechanistic understanding of these processes remains unknown. We have performed detailed pharmacological analyses of the role of the dynamic actin cytoskeleton in gravibending of maize (Zea mays) root apices. Depolymerization of actin filaments with two drugs having different mode of their actions, cytochalasin D and latrunculin B, stimulated root gravibending. By contrast, drug-induced stimulation of actin polymerization and inhibition of actin turnover, using two different agents phalloidin and jasplakinolide, compromised the root gravibending. Importantly, all these actin drugs inhibited root growth to similar extents suggesting that high actin turnover is essential for the gravity-related growth responses rather than for the general growth process. Both latrunculin B and cytochalasin D treatments inhibited root growth but restored gravibending of the decapped root apices, indicating that there is a strong potential for effective actin-mediated gravity sensing outside the cap. This elusive gravity sensing outside the root cap is dependent not only on the high rate of actin turnover but also on weakening of myosin activities, as general inhibition of myosin ATPases induced stimulation of gravibending of the decapped root apices. Collectively, these data provide evidence for the actin turnover-mediated gravity sensing outside the root cap. PMID:19521476

  20. KCC2 regulates actin dynamics in dendritic spines via interaction with β-PIX

    PubMed Central

    Llano, Olaya; Smirnov, Sergey; Soni, Shetal; Golubtsov, Andrey; Guillemin, Isabelle; Hotulainen, Pirta; Medina, Igor; Nothwang, Hans Gerd

    2015-01-01

    Chloride extrusion in mature neurons is largely mediated by the neuron-specific potassium-chloride cotransporter KCC2. In addition, independently of its chloride transport function, KCC2 regulates the development and morphology of dendritic spines through structural interactions with the actin cytoskeleton. The mechanism of this effect remains largely unknown. In this paper, we show a novel pathway for KCC2-mediated regulation of the actin cytoskeleton in neurons. We found that KCC2, through interaction with the b isoform of Rac/Cdc42 guanine nucleotide exchange factor β-PIX, regulates the activity of Rac1 GTPase and the phosphorylation of one of the major actin-regulating proteins, cofilin-1. KCC2-deficient neurons had abnormally high levels of phosphorylated cofilin-1. Consistently, dendritic spines of these neurons exhibited a large pool of stable actin, resulting in reduced spine motility and diminished density of functional synapses. In conclusion, we describe a novel signaling pathway that couples KCC2 to the cytoskeleton and regulates the formation of glutamatergic synapses. PMID:26056138

  1. The flatness of Lamellipodia explained by the interaction between actin dynamics and membrane deformation.

    PubMed

    Schmeiser, Christian; Winkler, Christoph

    2015-09-01

    The crawling motility of many cell types relies on lamellipodia, flat protrusions spreading on flat substrates but (on cells in suspension) also growing into three-dimensional space. Lamellipodia consist of a plasma membrane wrapped around an oriented actin filament meshwork. It is well known that the actin density is controlled by coordinated polymerization, branching, and capping processes, but the mechanisms producing the small aspect ratios of lamellipodia (hundreds of nm thickness vs. several μm lateral and inward extension) remain unclear. The main hypothesis of this work is a strong influence of the local geometry of the plasma membrane on the actin dynamics. This is motivated by observations of co-localization of proteins with I-BAR domains (like IRSp53) with polymerization and branching agents along the membrane. The I-BAR domains are known to bind to the membrane and to prefer and promote membrane curvature. This hypothesis is translated into a stochastic mathematical model where branching and capping rates, and polymerization speeds depend on the local membrane geometry and branching directions are influenced by the principal curvature directions. This requires the knowledge of the deformation of the membrane, being described in a quasi-stationary approximation by minimization of a modified Helfrich energy, subject to the actin filaments acting as obstacles. Simulations with this model predict pieces of flat lamellipodia without any prescribed geometric restrictions. PMID:26002996

  2. KCC2 regulates actin dynamics in dendritic spines via interaction with β-PIX.

    PubMed

    Llano, Olaya; Smirnov, Sergey; Soni, Shetal; Golubtsov, Andrey; Guillemin, Isabelle; Hotulainen, Pirta; Medina, Igor; Nothwang, Hans Gerd; Rivera, Claudio; Ludwig, Anastasia

    2015-06-01

    Chloride extrusion in mature neurons is largely mediated by the neuron-specific potassium-chloride cotransporter KCC2. In addition, independently of its chloride transport function, KCC2 regulates the development and morphology of dendritic spines through structural interactions with the actin cytoskeleton. The mechanism of this effect remains largely unknown. In this paper, we show a novel pathway for KCC2-mediated regulation of the actin cytoskeleton in neurons. We found that KCC2, through interaction with the b isoform of Rac/Cdc42 guanine nucleotide exchange factor β-PIX, regulates the activity of Rac1 GTPase and the phosphorylation of one of the major actin-regulating proteins, cofilin-1. KCC2-deficient neurons had abnormally high levels of phosphorylated cofilin-1. Consistently, dendritic spines of these neurons exhibited a large pool of stable actin, resulting in reduced spine motility and diminished density of functional synapses. In conclusion, we describe a novel signaling pathway that couples KCC2 to the cytoskeleton and regulates the formation of glutamatergic synapses. PMID:26056138

  3. Dissociation of F-actin induced by hydrostatic pressure.

    PubMed

    Garcia, C R; Amaral Júnior, J A; Abrahamsohn, P; Verjovski-Almeida, S

    1992-11-01

    F-actin purified from rabbit skeletal muscle undergoes reversible dissociation when subjected to hydrostatic pressures up to 240 MPa. Dissociation and reversibility were detected by the following procedures: fluorescence spectral changes observed under pressure, when either intrinsic tryptophan or pyrenyl emission of N-(1-pyrenyl)iodoacetamide-labeled actin were monitored; electron microscopy of samples fixed under pressure; size-exclusion HPLC of pressurized actin. The effect of pressure upon F-actin that had been polymerized in the presence of either Mg2+, Ca2+ or K+ was studied. The standard volume changes for the association of actin subunits, calculated from pressure/dissociation curves were 74 +/- 14 ml/mol for Mg-F-actin, 79 +/- 12 ml/mol for Ca-F-actin and 328 +/- 63 ml/mol for K-F-actin, indicating that actin subunits are packed differently in the polymer depending on which cation is present. All pressure/dissociation data could be fitted by a model for dissociation of a dimer, which suggests that in the F-actin filament there is a predominant intersubunit interaction interface, most likely the head-to-tail intrastrand interaction between two subunits which repeats itself along the polymer. A tenfold change in total protein concentration from 20 micrograms to 200 micrograms/ml Mg-F-actin did not cause a change in the pressure required for half-maximal dissociation. This indicates a heterogeneity of free energy of association among actin monomers in the Mg-F-actin polymer, suggesting that, in addition to the predominant intersubunit interaction, the disordered interactions in the filament significantly contribute to the heterogeneity of microenvironments in the interface between the subunits. PMID:1425683

  4. Fibroblast-mediated contraction in actinically exposed and actinically protected aging skin

    SciTech Connect

    Marks, M.W.; Morykwas, M.J.; Wheatley, M.J. )

    1990-08-01

    The changes in skin morphology over time are a consequence of both chronologic aging and the accumulation of environmental exposure. Through observation, we know that actinic radiation intensifies the apparent aging of skin. We have investigated the effects of aging and actinic radiation on the ability of fibroblasts to contract collagen-fibroblast lattices. Preauricular and postauricular skin samples were obtained from eight patients aged 49 to 74 undergoing rhytidectomy. The samples were kept separate, and the fibroblasts were grown in culture. Lattices constructed with preauricular fibroblasts consistently contracted more than lattices containing postauricular fibroblasts. The difference in amount of contraction in 7 days between sites was greatest for the younger patients and decreased linearly as donor age increased (r = -0.96). This difference may be due to preauricular fibroblasts losing their ability to contract a lattice as aging skin is exposed to more actinic radiation.

  5. Influence of YBa2HfO5.5 - 'derived secondary phase' on the critical current density and flux-Pinning force of YBa2Cu3O7-δ thick films

    NASA Astrophysics Data System (ADS)

    Rejith, Pullanhiyodan Puthiyaveedu; Vidya, Sukumariamma; Thomas, Jijimon Kumbukkattu

    2015-12-01

    Enhancement in critical current density (Jc) and flux pinning force (Fp) in superconducting thick films of YBa2Cu3O7-δ (YBCO) added with small quantities of nanopowders of HfO2, BaHfO3 and YBa2HfO5.5, coated on YBa2ZrO5.5 substrate by dip-coating technique is reported. Critical current density measurements were done over an applied magnetic field using standard four probe technique and the results are compared with that of pure YBCO. High critical current density (Jc) of ∼4.84 MA/cm2 at 77 K in self-field was obtained for 2 wt% of YBa2HfO5.5 added YBCO. A systematic increase in Jc observed in YBCO films prepared by the addition of nano HfO2, BaHfO3 and YBa2HfO5.5, attributed to the formation of a non-reacting 'derived secondary phase' YBa2HfO5.5 (YBHO) in the YBCO matrix. YBCO-YBa2HfO5.5 composite thick films have showed eightfold increases in Jc (3.29 MA/cm2) at 77 K and 0.4 T compared to pure YBa2Cu3O7-δ film (0.37 MA/cm2), while maintaining a high transition temperature (Tc). The development of effective pinning centers in nano particle added YBCO thick film have enhanced the flux pinning force from 1.8 GN/m3 for pure YBCO to a maximum value of 13.15 GN/m3 for YBCO-YBa2HfO5.5. X-ray diffraction and energy dispersive spectroscopic analysis confirmed the presence of secondary phase, derived in the matrix.

  6. Actin depolymerisation and crosslinking join forces with myosin II to contract actin coats on fused secretory vesicles.

    PubMed

    Miklavc, Pika; Ehinger, Konstantin; Sultan, Ayesha; Felder, Tatiana; Paul, Patrick; Gottschalk, Kay-Eberhard; Frick, Manfred

    2015-03-15

    In many secretory cells actin and myosin are specifically recruited to the surface of secretory granules following their fusion with the plasma membrane. Actomyosin-dependent compression of fused granules is essential to promote active extrusion of cargo. However, little is known about molecular mechanisms regulating actin coat formation and contraction. Here, we provide a detailed kinetic analysis of the molecules regulating actin coat contraction on fused lamellar bodies in primary alveolar type II cells. We demonstrate that ROCK1 and myosin light chain kinase 1 (MLCK1, also known as MYLK) translocate to fused lamellar bodies and activate myosin II on actin coats. However, myosin II activity is not sufficient for efficient actin coat contraction. In addition, cofilin-1 and α-actinin translocate to actin coats. ROCK1-dependent regulated actin depolymerisation by cofilin-1 in cooperation with actin crosslinking by α-actinin is essential for complete coat contraction. In summary, our data suggest a complementary role for regulated actin depolymerisation and crosslinking, and myosin II activity, to contract actin coats and drive secretion. PMID:25637593

  7. Actin-Dynamics in Plant Cells: The Function of Actin-Perturbing Substances: Jasplakinolide, Chondramides, Phalloidin, Cytochalasins, and Latrunculins.

    PubMed

    Holzinger, Andreas; Blaas, Kathrin

    2016-01-01

    This chapter gives an overview of the most common F-actin-perturbing substances that are used to study actin dynamics in living plant cells in studies on morphogenesis, motility, organelle movement, or when apoptosis has to be induced. These substances can be divided into two major subclasses: F-actin-stabilizing and -polymerizing substances like jasplakinolide and chondramides and F-actin-severing compounds like chytochalasins and latrunculins. Jasplakinolide was originally isolated form a marine sponge, and can now be synthesized and has become commercially available, which is responsible for its wide distribution as membrane-permeable F-actin-stabilizing and -polymerizing agent, which may even have anticancer activities. Cytochalasins, derived from fungi, show an F-actin-severing function and many derivatives are commercially available (A, B, C, D, E, H, J), also making it a widely used compound for F-actin disruption. The same can be stated for latrunculins (A, B), derived from red sea sponges; however the mode of action is different by binding to G-actin and inhibiting incorporation into the filament. In the case of swinholide a stable complex with actin dimers is formed resulting also in severing of F-actin. For influencing F-actin dynamics in plant cells only membrane permeable drugs are useful in a broad range. We however introduce also the phallotoxins and synthetic derivatives, as they are widely used to visualize F-actin in fixed cells. A particular uptake mechanism has been shown for hepatocytes, but has also been described in siphonal giant algae. In the present chapter the focus is set on F-actin dynamics in plant cells where alterations in cytoplasmic streaming can be particularly well studied; however methods by fluorescence applications including phalloidin and antibody staining as well as immunofluorescence-localization of the inhibitor drugs are given. PMID:26498789

  8. Actin-Dynamics in Plant Cells: The Function of Actin Perturbing Substances Jasplakinolide, Chondramides, Phalloidin, Cytochalasins, and Latrunculins

    PubMed Central

    Holzinger, Andreas; Blaas, Kathrin

    2016-01-01

    This chapter will give an overview of the most common F-actin perturbing substances, that are used to study actin dynamics in living plant cells in studies on morphogenesis, motility, organelle movement or when apoptosis has to be induced. These substances can be divided into two major subclasses – F-actin stabilizing and polymerizing substances like jasplakinolide, chondramides and F-actin severing compounds like chytochalasins and latrunculins. Jasplakinolide was originally isolated form a marine sponge, and can now be synthesized and has become commercially available, which is responsible for its wide distribution as membrane permeable F-actin stabilizing and polymerizing agent, which may even have anti-cancer activities. Cytochalasins, derived from fungi show an F-actin severing function and many derivatives are commercially available (A, B, C, D, E, H, J), also making it a widely used compound for F-actin disruption. The same can be stated for latrunculins (A, B), derived from red sea sponges, however the mode of action is different by binding to G-actin and inhibiting incorporation into the filament. In the case of swinholide a stable complex with actin dimers is formed resulting also in severing of F-actin. For influencing F-actin dynamics in plant cells only membrane permeable drugs are useful in a broad range. We however introduce also the phallotoxins and synthetic derivatives, as they are widely used to visualize F-actin in fixed cells. A particular uptake mechanism has been shown for hepatocytes, but has also been described in siphonal giant algae. In the present chapter the focus is set on F-actin dynamics in plant cells where alterations in cytoplasmic streaming can be particularly well studied; however methods by fluorescence applications including phalloidin- and antibody staining as well as immunofluorescence-localization of the inhibitor drugs are given. PMID:26498789

  9. Biases of CO2 Storage in Eddy Flux Measurements pertinent to Vertical Configurations of a Profile System and CO2 Density Averaging

    SciTech Connect

    Yang, Bai; Hanson, Paul J; Riggs, Jeffery S; Pallardy, Stephen G.; Hosman, K. P.; Meyers, T. P.; Wullschleger, Stan D; Gu, Lianhong; Heuer, Mark

    2007-01-01

    CO2 storage in a 30-minute period in a tall forest canopy often makes significant contributions to net ecosystem exchange (NEE) in the early morning and at night. When CO2 storage is properly measured and taken into account, underestimations of NEE on calm nights can be greatly reduced. Using CO2 data from a 12-level profile, we demonstrate that the lower canopy layer (below the thermal inversion) is a disproportional contributor to the total CO2 storage. This is because time derivative of CO2 density ( c/ t) generally shows increasing magnitude of mean and standard deviation with decreasing heights at night and from sunrise to 1000 hr in both growing and dormant seasons. Effects of resolution and configuration in a profiling system on the accuracy of CO2 storage estimation are evaluated by comparing subset profiles to the 12-level benchmark profile. It is demonstrated that the effectiveness of a profiling system in estimating CO2 storage is not only determined by its number of sampling levels but, more importantly, by its vertical configuration. To optimize a profile, one needs to balance the influence of two factors, c/ t and layer thickness, among all vertical sections within a forest. As a key contributor to the total CO2 storage, the lower canopy (with relatively large means and standard deviations of c/ t) requires a higher resolution in a profile system than the layers above. However, if the upper canopy is over-sparsely sampled relative to the lower canopy, the performance of a profile system might be degraded since, in such a situation, the influence of layer thickness dominates over that of c/ t. We also find that, because of different level of complexity in canopy structure, more sampling levels are necessary at our site in order to achieve the same level of accuracy as at a boreal aspen site. These results suggest that, in order to achieve an adequate accuracy in CO2 storage measurements, the number of sampling levels in a profile and its design should

  10. Spatial control of actin polymerization during neutrophil chemotaxis

    PubMed Central

    Weiner, Orion D.; Servant, Guy; Welch, Matthew D.; Mitchison, Timothy J.; Sedat, John W.; Bourne, Henry R.

    2010-01-01

    Neutrophils respond to chemotactic stimuli by increasing the nucleation and polymerization of actin filaments, but the location and regulation of these processes are not well understood. Here, using a permeabilized-cell assay, we show that chemotactic stimuli cause neutrophils to organize many discrete sites of actin polymerization, the distribution of which is biased by external chemotactic gradients. Furthermore, the Arp2/3 complex, which can nucleate actin polymerization, dynamically redistributes to the region of living neutrophils that receives maximal chemotactic stimulation, and the least-extractable pool of the Arp2/3 complex co-localizes with sites of actin polymerization. Our observations indicate that chemoattractant-stimulated neutrophils may establish discrete foci of actin polymerization that are similar to those generated at the posterior surface of the intracellular bacterium Listeria monocytogenes. We propose that asymmetrical establishment and/or maintenance of sites of actin polymerization produces directional migration of neutrophils in response to chemotactic gradients. PMID:10559877

  11. Identification of sucrose synthase as an actin-binding protein

    NASA Technical Reports Server (NTRS)

    Winter, H.; Huber, J. L.; Huber, S. C.; Davies, E. (Principal Investigator)

    1998-01-01

    Several lines of evidence indicate that sucrose synthase (SuSy) binds both G- and F-actin: (i) presence of SuSy in the Triton X-100-insoluble fraction of microsomal membranes (i.e. crude cytoskeleton fraction); (ii) co-immunoprecipitation of actin with anti-SuSy monoclonal antibodies; (iii) association of SuSy with in situ phalloidin-stabilized F-actin filaments; and (iv) direct binding to F-actin, polymerized in vitro. Aldolase, well known to interact with F-actin, interfered with binding of SuSy, suggesting that a common or overlapping binding site may be involved. We postulate that some of the soluble SuSy in the cytosol may be associated with the actin cytoskeleton in vivo.

  12. Photon flux requirements for EUV reticle imaging microscopy in the 22 and 16 nm nodes

    SciTech Connect

    Wintz, D.; Goldberg, K. A.; Mochi, I.; Huh, S.

    2010-03-12

    EUV-wavelength actinic microscopy yields detailed information about EUV mask patterns, architectures, defects, and the performance of defect repair strategies, without the complications of photoresist imaging. The measured aerial image intensity profiles provide valuable feedback to improve mask and lithography system modeling methods. In order to understand the photon-flux-dependent pattern measurement limits of EUV mask-imaging microscopy, we have investigated the effects of shot noise on aerial image linewidth measurements for lines in the 22 and 16-nm generations. Using a simple model of image formation near the resolution limit, we probe the influence of photon shot noise on the measured, apparent line roughness. With this methodology, we arrive at general flux density requirements independent of the specific EUV microscope configurations. Analytical and statistical analysis of aerial image simulations in the 22 and 16-nm generations reveal the trade-offs between photon energy density (controllable with exposure time), effective pixel dimension on the CCO (controlled by the microscope's magnification ratio), and image log slope (ILS). We find that shot-noise-induced linewidth roughness (LWR) varies imersely with the square root of the photon energy density, and is proportional to the imaging magnification ratio. While high magnification is necessary for adequate spatial resolution, for a given flux density, higher magnification ratios have diminishing benefits. With practical imaging parameters, we find that in order to achieve an LWR (3{sigma}) value of 5% of linewidth for dense, 88-nm mask features with 80% aerial image contrast and 13.5-nm effective pixel width (1000x magnification ratio), a peak photon flux of approximately 1400 photons per pixel per exposure is required.

  13. Scaling properties of the anisotropic critical current density in bulk textured YBaCuO. Evidence toward a 3D flux line lattice

    SciTech Connect

    Braithwaite, D.; Bourgault, D.; Sulpice, A.; Barbut, J.M.; Tournier, R. l'Universite Joseph Fourier, Grenoble ); Monot, I.; Lepropre, M.; Provost, J.; Desgardin, G. )

    1993-04-01

    The dc transport critical current densities of melt texture grown and magnetically melt textured bulk YBaCuO have been measured at 77 K and in magnetic fields. A maximum value of over 31,000 A/cm[sup 2] is obtained with a field of 7 teslas applied parallel to the (a,b) planes. Over the rest of the angular range the critical current is shown to be determined mainly by the c-axis component of the applied field. Although this dependency is expected in the presence of two-dimensional vortices, in fact the data are shown to correspond better to the behavior expected of an anisotropic three-dimensional superconductor. These results are compared to magnetization measurements on the same samples. Results show that when the field is directed close to the c-axis, superconducting transport currents flow at fields well above the field at which the irreversible magnetization disappears.

  14. Electrophoresis and orientation of F-actin in agarose gels.

    PubMed Central

    Borejdo, J; Ortega, H

    1989-01-01

    F-Actin was electrophoresed on agarose gels. In the presence of 2 mM MgCl2 and above pH 8.5 F-actin entered 1% agarose; when the electric field was 2.1 V/cm and the pH was 8.8, F-actin migrated through a gel as a single band at a rate of 2.5 mm/h. Labeling of actin with fluorophores did not affect its rate of migration, but an increase in ionic strength slowed it down. After the electrophoresis actin was able to bind phalloidin and heavy meromyosin (HMM) and it activated Mg2+-dependent ATPase activity of HMM. The mobility of F-actin increased with the rise in pH. Acto-S-1 complex was also able to migrate in agarose at basic pH, but at a lower rate than F-actin alone. The orientation of fluorescein labeled F-actin and of fluorescein labeled S-1 which formed rigor bonds with F-actin was measured during the electrophoresis by the fluorescence detected linear dichroism method. The former showed little orientation, probably because the dye was mobile on the surface of actin, but we were able to measure the orientation of the absorption dipole of the dye bound to S-1 which was attached to F-actin, and found that it assumed an orientation largely parallel to the direction of the electric field. These results show that actin can migrate in agarose gels in the F form and that it is oriented during the electrophoresis. Images FIGURE 1 FIGURE 3 FIGURE 4 PMID:2528384

  15. Measuring F-actin properties in dendritic spines

    PubMed Central

    Koskinen, Mikko; Hotulainen, Pirta

    2014-01-01

    During the last decade, numerous studies have demonstrated that the actin cytoskeleton plays a pivotal role in the control of dendritic spine shape. Synaptic stimulation rapidly changes the actin dynamics and many actin regulators have been shown to play roles in neuron functionality. Accordingly, defects in the regulation of the actin cytoskeleton in neurons have been implicated in memory disorders. Due to the small size of spines, it is difficult to detect changes in the actin structures in dendritic spines by conventional light microscopy imaging. Instead, to know how tightly actin filaments are bundled together, and how fast the filaments turnover, we need to use advanced microscopy techniques, such as fluorescence recovery after photobleaching (FRAP), photoactivatable green fluorescent protein (PAGFP) fluorescence decay and fluorescence anisotropy. Fluorescence anisotropy, which measures the Förster resonance energy transfer (FRET) between two GFP fluorophores, has been proposed as a method to measure the level of actin polymerization. Here, we propose a novel idea that fluorescence anisotropy could be more suitable to study the level of actin filament bundling instead of actin polymerization. We validate the method in U2OS cell line where the actin structures can be clearly distinguished and apply to analyze how actin filament organization in dendritic spines changes during neuronal maturation. In addition to fluorescence anisotropy validation, we take a critical look at the properties and limitations of FRAP and PAGFP fluorescence decay methods and offer our proposals for the analysis methods for these approaches. These three methods complement each other, each providing additional information about actin dynamics and organization in dendritic spines. PMID:25140131

  16. Ca2+-calmodulin regulates fesselin-induced actin polymerization.

    PubMed

    Schroeter, Mechthild; Chalovich, Joseph M

    2004-11-01

    Fesselin is a proline-rich actin-binding protein that was isolated from avian smooth muscle. Fesselin bundles actin and accelerates actin polymerization by facilitating nucleation. We now show that this polymerization of actin can be regulated by Ca(2+)-calmodulin. Fesselin was shown to bind to immobilized calmodulin in the presence of Ca(2+). The fesselin-calmodulin interaction was confirmed by a Ca(2+)-dependent increase in 2-(4-maleimidoanilino)naphthalene-6-sulfonic acid (MIANS) fluorescence upon addition of fesselin to MIANS-labeled wheat germ calmodulin. The affinity was estimated to be approximately 10(9) M(-1). The affinity of Ca(2+)-calmodulin to the fesselin F-actin complex was approximately 10(8) M(-1). Calmodulin binding to fesselin appeared to be functionally significant. In the presence of fesselin and calmodulin, the polymerization of actin was Ca(2+)-dependent. Ca(2+)-free calmodulin either had no effect or enhanced the ability of fesselin to accelerate actin polymerization. Ca(2+)-calmodulin not only reversed the stimulatory effect of fesselin but reduced the rate of polymerization below that observed in the absence of fesselin. While Ca(2+)-calmodulin had a large effect on the interaction of fesselin with G-actin, the effect on F-actin was small. Neither the binding of fesselin to F-actin nor the subsequent bundling of F-actin was greatly affected by Ca(2+)-calmodulin. Fesselin may function as an actin-polymerizing factor that is regulated by Ca(2+) levels. PMID:15504050

  17. A Robust Actin Filaments Image Analysis Framework.

    PubMed

    Alioscha-Perez, Mitchel; Benadiba, Carine; Goossens, Katty; Kasas, Sandor; Dietler, Giovanni; Willaert, Ronnie; Sahli, Hichem

    2016-08-01

    The cytoskeleton is a highly dynamical protein network that plays a central role in numerous cellular physiological processes, and is traditionally divided into three components according to its chemical composition, i.e. actin, tubulin and intermediate filament cytoskeletons. Understanding the cytoskeleton dynamics is of prime importance to unveil mechanisms involved in cell adaptation to any stress type. Fluorescence imaging of cytoskeleton structures allows analyzing the impact of mechanical stimulation in the cytoskeleton, but it also imposes additional challenges in the image processing stage, such as the presence of imaging-related artifacts and heavy blurring introduced by (high-throughput) automated scans. However, although there exists a considerable number of image-based analytical tools to address the image processing and analysis, most of them are unfit to cope with the aforementioned challenges. Filamentous structures in images can be considered as a piecewise composition of quasi-straight segments (at least in some finer or coarser scale). Based on this observation, we propose a three-steps actin filaments extraction methodology: (i) first the input image is decomposed into a 'cartoon' part corresponding to the filament structures in the image, and a noise/texture part, (ii) on the 'cartoon' image, we apply a multi-scale line detector coupled with a (iii) quasi-straight filaments merging algorithm for fiber extraction. The proposed robust actin filaments image analysis framework allows extracting individual filaments in the presence of noise, artifacts and heavy blurring. Moreover, it provides numerous parameters such as filaments orientation, position and length, useful for further analysis. Cell image decomposition is relatively under-exploited in biological images processing, and our study shows the benefits it provides when addressing such tasks. Experimental validation was conducted using publicly available datasets, and in osteoblasts grown in

  18. Mechanoaccumulative Elements of the Mammalian Actin Cytoskeleton.

    PubMed

    Schiffhauer, Eric S; Luo, Tianzhi; Mohan, Krithika; Srivastava, Vasudha; Qian, Xuyu; Griffis, Eric R; Iglesias, Pablo A; Robinson, Douglas N

    2016-06-01

    To change shape, divide, form junctions, and migrate, cells reorganize their cytoskeletons in response to changing mechanical environments [1-4]. Actin cytoskeletal elements, including myosin II motors and actin crosslinkers, structurally remodel and activate signaling pathways in response to imposed stresses [5-9]. Recent studies demonstrate the importance of force-dependent structural rearrangement of α-catenin in adherens junctions [10] and vinculin's molecular clutch mechanism in focal adhesions [11]. However, the complete landscape of cytoskeletal mechanoresponsive proteins and the mechanisms by which these elements sense and respond to force remain to be elucidated. To find mechanosensitive elements in mammalian cells, we examined protein relocalization in response to controlled external stresses applied to individual cells. Here, we show that non-muscle myosin II, α-actinin, and filamin accumulate to mechanically stressed regions in cells from diverse lineages. Using reaction-diffusion models for force-sensitive binding, we successfully predicted which mammalian α-actinin and filamin paralogs would be mechanoaccumulative. Furthermore, a "Goldilocks zone" must exist for each protein where the actin-binding affinity must be optimal for accumulation. In addition, we leveraged genetic mutants to gain a molecular understanding of the mechanisms of α-actinin and filamin catch-bonding behavior. Two distinct modes of mechanoaccumulation can be observed: a fast, diffusion-based accumulation and a slower, myosin II-dependent cortical flow phase that acts on proteins with specific binding lifetimes. Finally, we uncovered cell-type- and cell-cycle-stage-specific control of the mechanosensation of myosin IIB, but not myosin IIA or IIC. Overall, these mechanoaccumulative mechanisms drive the cell's response to physical perturbation during proper tissue development and disease. PMID:27185555

  19. A Robust Actin Filaments Image Analysis Framework

    PubMed Central

    Alioscha-Perez, Mitchel; Benadiba, Carine; Goossens, Katty; Kasas, Sandor; Dietler, Giovanni; Willaert, Ronnie; Sahli, Hichem

    2016-01-01

    The cytoskeleton is a highly dynamical protein network that plays a central role in numerous cellular physiological processes, and is traditionally divided into three components according to its chemical composition, i.e. actin, tubulin and intermediate filament cytoskeletons. Understanding the cytoskeleton dynamics is of prime importance to unveil mechanisms involved in cell adaptation to any stress type. Fluorescence imaging of cytoskeleton structures allows analyzing the impact of mechanical stimulation in the cytoskeleton, but it also imposes additional challenges in the image processing stage, such as the presence of imaging-related artifacts and heavy blurring introduced by (high-throughput) automated scans. However, although there exists a considerable number of image-based analytical tools to address the image processing and analysis, most of them are unfit to cope with the aforementioned challenges. Filamentous structures in images can be considered as a piecewise composition of quasi-straight segments (at least in some finer or coarser scale). Based on this observation, we propose a three-steps actin filaments extraction methodology: (i) first the input image is decomposed into a ‘cartoon’ part corresponding to the filament structures in the image, and a noise/texture part, (ii) on the ‘cartoon’ image, we apply a multi-scale line detector coupled with a (iii) quasi-straight filaments merging algorithm for fiber extraction. The proposed robust actin filaments image analysis framework allows extracting individual filaments in the presence of noise, artifacts and heavy blurring. Moreover, it provides numerous parameters such as filaments orientation, position and length, useful for further analysis. Cell image decomposition is relatively under-exploited in biological images processing, and our study shows the benefits it provides when addressing such tasks. Experimental validation was conducted using publicly available datasets, and in osteoblasts

  20. Visualization of actin filaments and monomers in somatic cell nuclei.

    PubMed

    Belin, Brittany J; Cimini, Beth A; Blackburn, Elizabeth H; Mullins, R Dyche

    2013-04-01

    In addition to its long-studied presence in the cytoplasm, actin is also found in the nuclei of eukaryotic cells. The function and form (monomer, filament, or noncanonical oligomer) of nuclear actin are hotly debated, and its localization and dynamics are largely unknown. To determine the distribution of nuclear actin in live somatic cells and evaluate its potential functions, we constructed and validated fluorescent nuclear actin probes. Monomeric actin probes concentrate in nuclear speckles, suggesting an interaction of monomers with RNA-processing factors. Filamentous actin probes recognize discrete structures with submicron lengths that are excluded from chromatin-rich regions. In time-lapse movies, these actin filament structures exhibit one of two types of mobility: 1) diffusive, with an average diffusion coefficient of 0.06-0.08 μm(2)/s, or (2) subdiffusive, with a mobility coefficient of 0.015 μm(2)/s. Individual filament trajectories exhibit features of particles moving within a viscoelastic mesh. The small size of nuclear actin filaments is inconsistent with a role in micron-scale intranuclear transport, and their localization suggests that they do not participate directly in chromatin-based processes. Our results instead suggest that actin filaments form part of a large, viscoelastic structure in the nucleoplasm and may act as scaffolds that help organize nuclear contents. PMID:23447706

  1. Human cytoplasmic actin proteins are encoded by a multigene family

    SciTech Connect

    Engel, J.; Gunning, P.; Kedes, L.

    1982-06-01

    The authors characterized nine human actin genes that they isolated from a library of cloned human DNA. Measurements of the thermal stability of hybrids formed between each cloned actin gene and ..cap alpha..-, ..beta..-, and ..gamma..-actin mRNA demonstrated that only one of the clones is most homologous to sarcomeric actin mRNA, whereas the remaining eight clones are most homologous to cytoplasmic actin mRNA. By the following criteria they show that these nine clones represent nine different actin gene loci rather than different alleles or different parts of a single gene: (i) the restriction enzyme maps of the coding regions are dissimilar; (ii) each clone contains sufficient coding region to encode all or most of an entire actin gene; and (iii) each clone contains sequences homologous to both the 5' and 3' ends of the coding region of a cloned chicken ..beta..-actin cDNA. They conclude, therefore, that the human cytoplasmic actin proteins are encoded by a multigene family.

  2. Myosin Vs organize actin cables in fission yeast

    PubMed Central

    Lo Presti, Libera; Chang, Fred; Martin, Sophie G.

    2012-01-01

    Myosin V motors are believed to contribute to cell polarization by carrying cargoes along actin tracks. In Schizosaccharomyces pombe, Myosin Vs transport secretory vesicles along actin cables, which are dynamic actin bundles assembled by the formin For3 at cell poles. How these flexible structures are able to extend longitudinally in the cell through the dense cytoplasm is unknown. Here we show that in myosin V (myo52 myo51) null cells, actin cables are curled, bundled, and fail to extend into the cell interior. They also exhibit reduced retrograde flow, suggesting that formin-mediated actin assembly is impaired. Myo52 may contribute to actin cable organization by delivering actin regulators to cell poles, as myoV∆ defects are partially suppressed by diverting cargoes toward cell tips onto microtubules with a kinesin 7–Myo52 tail chimera. In addition, Myo52 motor activity may pull on cables to provide the tension necessary for their extension and efficient assembly, as artificially tethering actin cables to the nuclear envelope via a Myo52 motor domain restores actin cable extension and retrograde flow in myoV mutants. Together these in vivo data reveal elements of a self-organizing system in which the motors shape their own tracks by transporting cargoes and exerting physical pulling forces. PMID:23051734

  3. Force Generation by Endocytic Actin Patches in Budding Yeast

    PubMed Central

    Carlsson, Anders E.; Bayly, Philip V.

    2014-01-01

    Membrane deformation during endocytosis in yeast is driven by local, templated assembly of a sequence of proteins including polymerized actin and curvature-generating coat proteins such as clathrin. Actin polymerization is required for successful endocytosis, but it is not known by what mechanisms actin polymerization generates the required pulling forces. To address this issue, we develop a simulation method in which the actin network at the protein patch is modeled as an active gel. The deformation of the gel is treated using a finite-element approach. We explore the effects and interplay of three different types of force driving invagination: 1), forces perpendicular to the membrane, generated by differences between actin polymerization rates at the edge of the patch and those at the center; 2), the inherent curvature of the coat-protein layer; and 3), forces parallel to the membrane that buckle the coat protein layer, generated by an actomyosin contractile ring. We find that with optimistic estimates for the stall stress of actin gel growth and the shear modulus of the actin gel, actin polymerization can generate almost enough force to overcome the turgor pressure. In combination with the other mechanisms, actin polymerization can the force over the critical value. PMID:24739159

  4. Myosin motors fragment and compact membrane-bound actin filaments

    PubMed Central

    Vogel, Sven K; Petrasek, Zdenek; Heinemann, Fabian; Schwille, Petra

    2013-01-01

    Cell cortex remodeling during cell division is a result of myofilament-driven contractility of the cortical membrane-bound actin meshwork. Little is known about the interaction between individual myofilaments and membrane-bound actin filaments. Here we reconstituted a minimal actin cortex to directly visualize the action of individual myofilaments on membrane-bound actin filaments using TIRF microscopy. We show that synthetic myofilaments fragment and compact membrane-bound actin while processively moving along actin filaments. We propose a mechanism by which tension builds up between the ends of myofilaments, resulting in compressive stress exerted to single actin filaments, causing their buckling and breakage. Modeling of this mechanism revealed that sufficient force (∼20 pN) can be generated by single myofilaments to buckle and break actin filaments. This mechanism of filament fragmentation and compaction may contribute to actin turnover and cortex reorganization during cytokinesis. DOI: http://dx.doi.org/10.7554/eLife.00116.001 PMID:23326639

  5. An unconventional form of actin in protozoan hemoflagellate, Leishmania.

    PubMed

    Kapoor, Prabodh; Sahasrabuddhe, Amogh A; Kumar, Ashutosh; Mitra, Kalyan; Siddiqi, Mohammad Imran; Gupta, Chhitar M

    2008-08-15

    Leishmania actin was cloned, overexpressed in baculovirus-insect cell system, and purified to homogeneity. The purified protein polymerized optimally in the presence of Mg2+ and ATP, but differed from conventional actins in its following properties: (i) it did not polymerize in the presence of Mg2+ alone, (ii) it polymerized in a restricted range of pH 7.0-8.5, (iii) its critical concentration for polymerization was found to be 3-4-fold lower than of muscle actin, (iv) it predominantly formed bundles rather than single filaments at pH 8.0, (v) it displayed considerably higher ATPase activity during polymerization, (vi) it did not inhibit DNase-I activity, and (vii) it did not bind the F-actin-binding toxin phalloidin or the actin polymerization disrupting agent Latrunculin B. Computational and molecular modeling studies revealed that the observed unconventional behavior of Leishmania actin is related to the diverged amino acid stretches in its sequence, which may lead to changes in the overall charge distribution on its solvent-exposed surface, ATP binding cleft, Mg2+ binding sites, and the hydrophobic loop that is involved in monomer-monomer interactions. Phylogenetically, it is related to ciliate actins, but to the best of our knowledge, no other actin with such unconventional properties has been reported to date. It is therefore suggested that actin in Leishmania may serve as a novel target for design of new antileishmanial drugs. PMID:18539603

  6. VASP Governs Actin Dynamics by Modulating Filament Anchoring

    PubMed Central

    Trichet, Léa; Campàs, Otger; Sykes, Cécile; Plastino, Julie

    2007-01-01

    Actin filament dynamics at the cell membrane are important for cell-matrix and cell-cell adhesions and the protrusion of the leading edge. Since actin filaments must be connected to the cell membrane to exert forces but must also detach from the membrane to allow it to move and evolve, the balance between actin filament tethering and detachment at adhesion sites and the leading edge is key for cell shape changes and motility. How this fine tuning is performed in cells remains an open question, but possible candidates are the Drosophila enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) family of proteins, which localize to dynamic actin structures in the cell. Here we study VASP-mediated actin-related proteins 2/3 (Arp2/3) complex-dependent actin dynamics using a substrate that mimics the fluid properties of the cell membrane: an oil-water interface. We show evidence that polymerization activators undergo diffusion and convection on the fluid surface, due to continual attachment and detachment to the actin network. These dynamics are enhanced in the presence of VASP, and we observe cycles of catastrophic detachment of the actin network from the surface, resulting in stop-and-go motion. These results point to a role for VASP in the modulation of filament anchoring, with implications for actin dynamics at cell adhesions and at the leading edge of the cell. PMID:17098798

  7. Solubilization of native actin monomers from human erythrocyte membranes.

    PubMed

    Tilley, L; Dwyer, M; Ralston, G B

    1986-01-01

    Up to 50% of the actin in erythrocyte membranes can be solubilized at low ionic strength in a form capable of inhibiting DNAse I, in the presence of 0.4 mM ATP and 0.05 mM calcium. In the absence of calcium and ATP, actin is released but is apparently rapidly denatured. Solubilization of G-actin increases with temperature up to 37 degrees C. At higher temperatures, actin is released rapidly but quickly loses its ability to inhibit DNAse I. PMID:3789986

  8. Electrostatics control actin filament nucleation and elongation kinetics.

    PubMed

    Crevenna, Alvaro H; Naredi-Rainer, Nikolaus; Schönichen, André; Dzubiella, Joachim; Barber, Diane L; Lamb, Don C; Wedlich-Söldner, Roland

    2013-04-26

    The actin cytoskeleton is a central mediator of cellular morphogenesis, and rapid actin reorganization drives essential processes such as cell migration and cell division. Whereas several actin-binding proteins are known to be regulated by changes in intracellular pH, detailed information regarding the effect of pH on the actin dynamics itself is still lacking. Here, we combine bulk assays, total internal reflection fluorescence microscopy, fluorescence fluctuation spectroscopy techniques, and theory to comprehensively characterize the effect of pH on actin polymerization. We show that both nucleation and elongation are strongly enhanced at acidic pH, with a maximum close to the pI of actin. Monomer association rates are similarly affected by pH at both ends, although dissociation rates are differentially affected. This indicates that electrostatics control the diffusional encounter but not the dissociation rate, which is critical for the establishment of actin filament asymmetry. A generic model of protein-protein interaction, including electrostatics, explains the observed pH sensitivity as a consequence of charge repulsion. The observed pH effect on actin in vitro agrees with measurements of Listeria propulsion in pH-controlled cells. pH regulation should therefore be considered as a modulator of actin dynamics in a cellular environment. PMID:23486468

  9. Helical buckling of actin inside filopodia generates traction

    PubMed Central

    Leijnse, Natascha; Oddershede, Lene B.; Bendix, Poul M.

    2015-01-01

    Cells can interact with their surroundings via filopodia, which are membrane protrusions that extend beyond the cell body. Filopodia are essential during dynamic cellular processes like motility, invasion, and cell–cell communication. Filopodia contain cross-linked actin filaments, attached to the surrounding cell membrane via protein linkers such as integrins. These actin filaments are thought to play a pivotal role in force transduction, bending, and rotation. We investigated whether, and how, actin within filopodia is responsible for filopodia dynamics by conducting simultaneous force spectroscopy and confocal imaging of F-actin in membrane protrusions. The actin shaft was observed to periodically undergo helical coiling and rotational motion, which occurred simultaneously with retrograde movement of actin inside the filopodium. The cells were found to retract beads attached to the filopodial tip, and retraction was found to correlate with rotation and coiling of the actin shaft. These results suggest a previously unidentified mechanism by which a cell can use rotation of the filopodial actin shaft to induce coiling and hence axial shortening of the filopodial actin bundle. PMID:25535347

  10. Myosin Vs organize actin cables in fission yeast.

    PubMed

    Lo Presti, Libera; Chang, Fred; Martin, Sophie G

    2012-12-01

    Myosin V motors are believed to contribute to cell polarization by carrying cargoes along actin tracks. In Schizosaccharomyces pombe, Myosin Vs transport secretory vesicles along actin cables, which are dynamic actin bundles assembled by the formin For3 at cell poles. How these flexible structures are able to extend longitudinally in the cell through the dense cytoplasm is unknown. Here we show that in myosin V (myo52 myo51) null cells, actin cables are curled, bundled, and fail to extend into the cell interior. They also exhibit reduced retrograde flow, suggesting that formin-mediated actin assembly is impaired. Myo52 may contribute to actin cable organization by delivering actin regulators to cell poles, as myoV defects are partially suppressed by diverting cargoes toward cell tips onto microtubules with a kinesin 7-Myo52 tail chimera. In addition, Myo52 motor activity may pull on cables to provide the tension necessary for their extension and efficient assembly, as artificially tethering actin cables to the nuclear envelope via a Myo52 motor domain restores actin cable extension and retrograde flow in myoV mutants. Together these in vivo data reveal elements of a self-organizing system in which the motors shape their own tracks by transporting cargoes and exerting physical pulling forces. PMID:23051734

  11. Dependence of the flux-creep activation energy on current density and magnetic field for a Ca10(Pt3As8)[(Fe1-xPtx)2As2]5 single crystal

    NASA Astrophysics Data System (ADS)

    Ge, J.; Gutierrez, J.; Li, J.; Yuan, J.; Wang, H.-B.; Yamaura, K.; Takayama-Muromachi, E.; Moshchalkov, V. V.

    2014-03-01

    We have performed detailed ac susceptibility measurements to investigate the vortex dynamics in a Ca10(Pt3As8)[(Fe1-xPtx)2As2]5 single crystal as a function of temperature, frequency, ac amplitude, and dc field. The field dependence of the activation energy U is derived in the framework of thermally activated flux creep theory, yielding a power law dependence of U ˜ Hα with α ≈ -1.0 for H above 0.30 T, while below 0.3 T U is independent of the field. The activation energy reaches 104 K at low fields, suggesting strong pinning in the material. The nonlinear function of the activation energy vs. the current density is determined, which shows logarithmic dependence U(J)∝lnJ.

  12. Influence of random point defects introduced by proton irradiation on the flux creep rates and magnetic field dependence of the critical current density J c of co-evaporated GdBa2Cu3O7-δ coated conductors

    NASA Astrophysics Data System (ADS)

    Haberkorn, N.; Kim, Jeehoon; Suárez, S.; Lee, Jae-Hun; Moon, S. H.

    2015-12-01

    We report the influence of random point defects introduced by 3 MeV proton irradiation (doses of 0.5 × 1016, 1 × 1016, 2 × 1016 and 6 × 1016 cm-2) on the vortex dynamics of co-evaporated 1.3 μm thick, GdBa2Cu3O7-δ coated conductors. Our results indicate that the inclusion of additional random point defects reduces the low field and enhances the in-field critical current densities J c. The main in-field J c enhancement takes place below 40 K, which is in agreement with the expectations for pinning by random point defects. In addition, our data show a slight though clear increase in flux creep rates as a function of irradiation fluence. Maley analysis indicates that this increment can be associated with a reduction in the exponent μ characterizing the glassy behavior.

  13. Calcium and actin in the saga of awakening oocytes.

    PubMed

    Santella, Luigia; Limatola, Nunzia; Chun, Jong T

    2015-04-24

    The interaction of the spermatozoon with the egg at fertilization remains one of the most fascinating mysteries of life. Much of our scientific knowledge on fertilization comes from studies on sea urchin and starfish, which provide plenty of gametes. Large and transparent, these eggs have served as excellent model systems for studying egg activation and embryo development in seawater, a plain natural medium. Starfish oocytes allow the study of the cortical, cytoplasmic and nuclear changes during the meiotic maturation process, which can also be triggered in vitro by hormonal stimulation. These morphological and biochemical changes ensure successful fertilization of the eggs at the first metaphase. On the other hand, sea urchin eggs are fertilized after the completion of meiosis, and are particularly suitable for the study of sperm-egg interaction, early events of egg activation, and embryonic development, as a large number of mature eggs can be fertilized synchronously. Starfish and sea urchin eggs undergo abrupt changes in the cytoskeleton and ion fluxes in response to the fertilizing spermatozoon. The plasma membrane and cortex of an egg thus represent "excitable media" that quickly respond to the stimulus with the Ca(2+) swings and structural changes. In this article, we review some of the key findings on the rapid dynamic rearrangements of the actin cytoskeleton in the oocyte/egg cortex upon hormonal or sperm stimulation and their roles in the modulation of the Ca(2+) signals and in the control of monospermic fertilization. PMID:25998739

  14. Conical electromagnetic radiation flux concentrator

    NASA Technical Reports Server (NTRS)

    Miller, E. R.

    1972-01-01

    Concentrator provides method of concentrating a beam of electromagnetic radiation into a smaller beam, presenting a higher flux density. Smaller beam may be made larger by sending radiation through the device in the reverse direction.

  15. The Pseudomonas syringae Type III Effector HopG1 Induces Actin Remodeling to Promote Symptom Development and Susceptibility during Infection1[OPEN

    PubMed Central

    Shimono, Masaki; Porter, Katie; Kvitko, Brian H.; Henty-Ridilla, Jessica; Creason, Allison; Chang, Jeff H.; Staiger, Christopher J.

    2016-01-01

    The plant cytoskeleton underpins the function of a multitude of cellular mechanisms, including those associated with developmental- and stress-associated signaling processes. In recent years, the actin cytoskeleton has been demonstrated to play a key role in plant immune signaling, including a recent demonstration that pathogens target actin filaments to block plant defense and immunity. Herein, we quantified spatial changes in host actin filament organization after infection with Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), demonstrating that the type-III effector HopG1 is required for pathogen-induced changes to actin filament architecture and host disease symptom development during infection. Using a suite of pathogen effector deletion constructs, coupled with high-resolution microscopy, we found that deletion of hopG1 from Pst DC3000 resulted in a reduction in actin bundling and a concomitant increase in the density of filament arrays in Arabidopsis, both of which correlate with host disease symptom development. As a mechanism underpinning this activity, we further show that the HopG1 effector interacts with an Arabidopsis mitochondrial-localized kinesin motor protein. Kinesin mutant plants show reduced disease symptoms after pathogen infection, which can be complemented by actin-modifying agents. In total, our results support a model in which HopG1 induces changes in the organization of the actin cytoskeleton as part of its virulence function in promoting disease symptom development. PMID:27217495

  16. The Pseudomonas syringae Type III Effector HopG1 Induces Actin Remodeling to Promote Symptom Development and Susceptibility during Infection.

    PubMed

    Shimono, Masaki; Lu, Yi-Ju; Porter, Katie; Kvitko, Brian H; Henty-Ridilla, Jessica; Creason, Allison; He, Sheng Yang; Chang, Jeff H; Staiger, Christopher J; Day, Brad

    2016-07-01

    The plant cytoskeleton underpins the function of a multitude of cellular mechanisms, including those associated with developmental- and stress-associated signaling processes. In recent years, the actin cytoskeleton has been demonstrated to play a key role in plant immune signaling, including a recent demonstration that pathogens target actin filaments to block plant defense and immunity. Herein, we quantified spatial changes in host actin filament organization after infection with Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), demonstrating that the type-III effector HopG1 is required for pathogen-induced changes to actin filament architecture and host disease symptom development during infection. Using a suite of pathogen effector deletion constructs, coupled with high-resolution microscopy, we found that deletion of hopG1 from Pst DC3000 resulted in a reduction in actin bundling and a concomitant increase in the density of filament arrays in Arabidopsis, both of which correlate with host disease symptom development. As a mechanism underpinning this activity, we further show that the HopG1 effector interacts with an Arabidopsis mitochondrial-localized kinesin motor protein. Kinesin mutant plants show reduced disease symptoms after pathogen infection, which can be complemented by actin-modifying agents. In total, our results support a model in which HopG1 induces changes in the organization of the actin cytoskeleton as part of its virulence function in promoting disease symptom development. PMID:27217495

  17. A small molecule inhibitor of tropomyosin dissociates actin binding from tropomyosin-directed regulation of actin dynamics

    PubMed Central

    Bonello, Teresa T.; Janco, Miro; Hook, Jeff; Byun, Alex; Appaduray, Mark; Dedova, Irina; Hitchcock-DeGregori, Sarah; Hardeman, Edna C.; Stehn, Justine R.; Böcking, Till; Gunning, Peter W.

    2016-01-01

    The tropomyosin family of proteins form end-to-end polymers along the actin filament. Tumour cells rely on specific tropomyosin-containing actin filament populations for growth and survival. To dissect out the role of tropomyosin in actin filament regulation we use the small molecule TR100 directed against the C terminus of the tropomyosin isoform Tpm3.1. TR100 nullifies the effect of Tpm3.1 on actin depolymerisation but surprisingly Tpm3.1 retains the capacity to bind F-actin in a cooperative manner. In vivo analysis also confirms that, in the presence of TR100, fluorescently tagged Tpm3.1 recovers normally into stress fibers. Assembling end-to-end along the actin filament is thereby not sufficient for tropomyosin to fulfil its function. Rather, regulation of F-actin stability by tropomyosin requires fidelity of information communicated at the barbed end of the actin filament. This distinction has significant implications for perturbing tropomyosin-dependent actin filament function in the context of anti-cancer drug development. PMID:26804624

  18. CASEIN KINASE1-LIKE PROTEIN2 Regulates Actin Filament Stability and Stomatal Closure via Phosphorylation of Actin Depolymerizing Factor.

    PubMed

    Zhao, Shuangshuang; Jiang, Yuxiang; Zhao, Yang; Huang, Shanjin; Yuan, Ming; Zhao, Yanxiu; Guo, Yan

    2016-06-01

    The opening and closing of stomata are crucial for plant photosynthesis and transpiration. Actin filaments undergo dynamic reorganization during stomatal closure, but the underlying mechanism for this cytoskeletal reorganization remains largely unclear. In this study, we identified and characterized Arabidopsis thaliana casein kinase 1-like protein 2 (CKL2), which responds to abscisic acid (ABA) treatment and participates in ABA- and drought-induced stomatal closure. Although CKL2 does not bind to actin filaments directly and has no effect on actin assembly in vitro, it colocalizes with and stabilizes actin filaments in guard cells. Further investigation revealed that CKL2 physically interacts with and phosphorylates actin depolymerizing factor 4 (ADF4) and inhibits its activity in actin filament disassembly. During ABA-induced stomatal closure, deletion of CKL2 in Arabidopsis alters actin reorganization in stomata and renders stomatal closure less sensitive to ABA, whereas deletion of ADF4 impairs the disassembly of actin filaments and causes stomatal closure to be more sensitive to ABA Deletion of ADF4 in the ckl2 mutant partially recues its ABA-insensitive stomatal closure phenotype. Moreover, Arabidopsis ADFs from subclass I are targets of CKL2 in vitro. Thus, our results suggest that CKL2 regulates actin filament reorganization and stomatal closure mainly through phosphorylation of ADF. PMID:27268429

  19. Actin turnover-dependent fast dissociation of capping protein in the dendritic nucleation actin network: evidence of frequent filament severing.

    PubMed

    Miyoshi, Takushi; Tsuji, Takahiro; Higashida, Chiharu; Hertzog, Maud; Fujita, Akiko; Narumiya, Shuh; Scita, Giorgio; Watanabe, Naoki

    2006-12-18

    Actin forms the dendritic nucleation network and undergoes rapid polymerization-depolymerization cycles in lamellipodia. To elucidate the mechanism of actin disassembly, we characterized molecular kinetics of the major filament end-binding proteins Arp2/3 complex and capping protein (CP) using single-molecule speckle microscopy. We have determined the dissociation rates of Arp2/3 and CP as 0.048 and 0.58 s(-1), respectively, in lamellipodia of live XTC fibroblasts. This CP dissociation rate is three orders of magnitude faster than in vitro. CP dissociates slower from actin stress fibers than from the lamellipodial actin network, suggesting that CP dissociation correlates with actin filament dynamics. We found that jasplakinolide, an actin depolymerization inhibitor, rapidly blocked the fast CP dissociation in cells. Consistently, the coexpression of LIM kinase prolonged CP speckle lifetime in lamellipodia. These results suggest that cofilin-mediated actin disassembly triggers CP dissociation from actin filaments. We predict that filament severing and end-to-end annealing might take place fairly frequently in the dendritic nucleation actin arrays. PMID:17178911

  20. Differential Actin-regulatory Activities of Tropomodulin1 and Tropomodulin3 with Diverse Tropomyosin and Actin Isoforms*

    PubMed Central

    Yamashiro, Sawako; Gokhin, David S.; Sui, Zhenhua; Bergeron, Sarah E.; Rubenstein, Peter A.; Fowler, Velia M.

    2014-01-01

    Tropomodulins (Tmods) are F-actin pointed end capping proteins that interact with tropomyosins (TMs) and cap TM-coated filaments with higher affinity than TM-free filaments. Here, we tested whether differences in recognition of TM or actin isoforms by Tmod1 and Tmod3 contribute to the distinct cellular functions of these Tmods. We found that Tmod3 bound ∼5-fold more weakly than Tmod1 to α/βTM, TM5b, and TM5NM1. However, surprisingly, Tmod3 was as effective as Tmod1 at capping pointed ends of skeletal muscle α-actin (αsk-actin) filaments coated with α/βTM, TM5b, or TM5NM1. Tmod3 only capped TM-coated αsk-actin filaments more weakly than Tmod1 in the presence of recombinant αTM2, which is unacetylated at its NH2 terminus, binds F-actin weakly, and has a disabled Tmod-binding site. Moreover, both Tmod1 and Tmod3 were similarly effective at capping pointed ends of platelet β/cytoplasmic γ (γcyto)-actin filaments coated with TM5NM1. In the absence of TMs, both Tmod1 and Tmod3 had similarly weak abilities to nucleate β/γcyto-actin filament assembly, but only Tmod3 could sequester cytoplasmic β- and γcyto-actin (but not αsk-actin) monomers and prevent polymerization under physiological conditions. Thus, differences in TM binding by Tmod1 and Tmod3 do not appear to regulate the abilities of these Tmods to cap TM-αsk-actin or TM-β/γcyto-actin pointed ends and, thus, are unlikely to determine selective co-assembly of Tmod, TM, and actin isoforms in different cell types and cytoskeletal structures. The ability of Tmod3 to sequester β- and γcyto-actin (but not αsk-actin) monomers in the absence of TMs suggests a novel function for Tmod3 in regulating actin remodeling or turnover in cells. PMID:24644292

  1. Evidence That an Unconventional Actin Can Provide Essential F-Actin Function and That a Surveillance System Monitors F-Actin Integrity in Chlamydomonas.

    PubMed

    Onishi, Masayuki; Pringle, John R; Cross, Frederick R

    2016-03-01

    Actin is one of the most conserved eukaryotic proteins. It is thought to have multiple essential cellular roles and to function primarily or exclusively as filaments ("F-actin"). Chlamydomonas has been an enigma, because a null mutation (ida5-1) in its single gene for conventional actin does not affect growth. A highly divergent actin gene, NAP1, is upregulated in ida5-1 cells, but it has been unclear whether NAP1 can form filaments or provide actin function. Here, we used the actin-depolymerizing drug latrunculin B (LatB), the F-actin-specific probe Lifeact-Venus, and genetic and molecular methods to resolve these issues. LatB-treated wild-type cells continue to proliferate; they initially lose Lifeact-stained structures but recover them concomitant with upregulation of NAP1. Thirty-nine LatB-sensitive mutants fell into four genes (NAP1 and LAT1-LAT3) in which we identified the causative mutations using a novel combinatorial pool-sequencing strategy. LAT1-LAT3 are required for NAP1 upregulation upon LatB treatment, and ectopic expression of NAP1 largely rescues the LatB sensitivity of the lat1-lat3 mutants, suggesting that the LAT gene products comprise a regulatory hierarchy with NAP1 expression as the major functional output. Selection of LatB-resistant revertants of a nap1 mutant yielded dominant IDA5 mutations that presumably render F-IDA5 resistant to LatB, and nap1 and lat mutations are synthetically lethal with ida5-1 in the absence of LatB. We conclude that both IDA5 and the divergent NAP1 can form filaments and redundantly provide essential F-actin functions and that a novel surveillance system, probably responding to a loss of F-actin, triggers NAP1 expression and perhaps other compensatory responses. PMID:26715672

  2. Encoding Mechano-Memories in Actin Networks

    NASA Astrophysics Data System (ADS)

    Foucard, Louis; Majumdar, Sayantan; Levine, Alex; Gardel, Margaret

    The ability of cells to sense and adapt to external mechanical stimuli is vital to many of its biological functions. A critical question is therefore to understand how mechanosensory mechanisms arise in living matter, with implications in both cell biology and smart materials design. Experimental work has demonstrated that the mechanical properties of semiflexible actin networks in Eukaryotic cells can be modulated (either transiently or irreversibly) via the application of external forces. Previous work has also shown with a combination of numerical simulations and analytic calculations shows that the broken rotational symmetry of the filament orientational distribution in semiflexible networks leads to dramatic changes in the mechanical response. Here we demonstrate with a combination of numerical and analytic calculations that the observed long-lived mechano-memory in the actin networks arise from changes in the nematic order of the constituent filaments. These stress-induced changes in network topology relax slowly under zero stress and can be observed through changes in the nonlinear mechanics. Our results provide a strategy for designing a novel class of materials and demonstrate a new putative mechanism of mechanical sensing in eukaryotic cells.

  3. Force Transmission in the Actin Cytoskeleton

    NASA Astrophysics Data System (ADS)

    Gardel, Margaret

    2012-02-01

    The ability of cells to sense and generate mechanical forces is essential to numerous aspects of their physiology, including adhesion, migration, division and differentiation. To a large degree, cellular tension is regulated by the transmission of myosin II-generated forces through the filamentous actin (F-actin) cytoskeleton. While transmission of myosin-generated stresses from the molecular to cellular length scale is well understood in the context of highly organized sarcomeres found in striated muscle, non-muscle and smooth muscle cells contain a wide variety of bundles and networks lacking sarcomeric organization. I will describe the in vitro and in vivo approaches we use to study force transmission in such disordered actomyosin assemblies. Our in vivo results are showing that highly organized stress fibers contribute surprisingly little to the overall extent of cellular tension as compared to disordered actomyosin meshworks. Our in vitro results are demonstrating the mechanisms of symmetry breaking in disordered actomyosin bundles that facilitate the formation of contractile bundles with well-defined ``contractile elements.'' These results provide insight into the self-organization of actomyosin cytoskeleton in non-muscle cells that regulate and maintain cellular tension.

  4. Actin and Endocytosis in Budding Yeast

    PubMed Central

    Goode, Bruce L.; Eskin, Julian A.; Wendland, Beverly

    2015-01-01

    Endocytosis, the process whereby the plasma membrane invaginates to form vesicles, is essential for bringing many substances into the cell and for membrane turnover. The mechanism driving clathrin-mediated endocytosis (CME) involves > 50 different protein components assembling at a single location on the plasma membrane in a temporally ordered and hierarchal pathway. These proteins perform precisely choreographed steps that promote receptor recognition and clustering, membrane remodeling, and force-generating actin-filament assembly and turnover to drive membrane invagination and vesicle scission. Many critical aspects of the CME mechanism are conserved from yeast to mammals and were first elucidated in yeast, demonstrating that it is a powerful system for studying endocytosis. In this review, we describe our current mechanistic understanding of each step in the process of yeast CME, and the essential roles played by actin polymerization at these sites, while providing a historical perspective of how the landscape has changed since the preceding version of the YeastBook was published 17 years ago (1997). Finally, we discuss the key unresolved issues and where future studies might be headed. PMID:25657349

  5. How cofilin severs an actin filament.

    PubMed

    De La Cruz, Enrique M

    2009-05-15

    The actin regulatory protein, cofilin, promotes actin assembly dynamics by severing filaments and increasing the number of ends from which subunits add and dissociate. Recent studies provide biophysical descriptions of cooperative filament interactions in energetic, mechanical and structural terms. A one-dimensional Ising model with nearest-neighbor interactions permits thermodynamic analysis of cooperative binding and indicates that one or a few cofilin molecules can sever a filament. Binding and cooperative interactions are entropically driven. A significant fraction of the binding free energy results from the linked dissociation of filament-associated ions (polyelectrolyte effect), which modulate filament structure, stability and mechanics. The remaining binding free energy and essentially all of the cooperative free energy arise from the enhanced conformational dynamics of the cofilactin complex. Filament mechanics are modulated by cofilin such that cofilin-saturated filaments are approximately 10- to 20-fold more compliant in bending and twisting than bare filaments. Cofilin activity is well described by models in which discontinuities in topology, mechanics and conformational dynamics generate stress concentration and promote fracture at junctions of bare and decorated segments, analogous to the grain boundary fracture of crystalline materials and the thermally driven formation of shear transformation zones in colloidal glass. PMID:20700473

  6. Actin binding proteins, spermatid transport and spermiation*

    PubMed Central

    Qian, Xiaojing; Mruk, Dolores D.; Cheng, Yan-Ho; Tang, Elizabeth I.; Han, Daishu; Lee, Will M.; Wong, Elissa W. P.; Cheng, C. Yan

    2014-01-01

    The transport of germ cells across the seminiferous epithelium is composed of a series of cellular events during the epithelial cycle essential to the completion of spermatogenesis. Without the timely transport of spermatids during spermiogenesis, spermatozoa that are transformed from step 19 spermatids in the rat testis fail to reach the luminal edge of the apical compartment and enter the tubule lumen at spermiation, thereby entering the epididymis for further maturation. Step 19 spermatids and/or sperms that remain in the epithelium will be removed by the Sertoli cell via phagocytosis to form phagosomes and be degraded by lysosomes, leading to subfertility and/or infertility. However, the biology of spermatid transport, in particular the final events that lead to spermiation remain elusive. Based on recent data in the field, we critically evaluate the biology of spermiation herein by focusing on the actin binding proteins (ABPs) that regulate the organization of actin microfilaments at the Sertoli-spermatid interface, which is crucial for spermatid transport during this event. The hypothesis we put forth herein also highlights some specific areas of research that can be pursued by investigators in the years to come. PMID:24735648

  7. Drebrin inhibits cofilin-induced severing of F-actin.

    PubMed

    Grintsevich, Elena E; Reisler, Emil

    2014-08-01

    Molecular cross-talk between neuronal drebrin A and cofilin is believed to be a part of the activity-dependent cytoskeleton-modulating pathway in dendritic spines. Impairments in this pathway are implicated also in synaptic dysfunction in Alzheimer's disease, Down syndrome, epilepsy, and normal aging. However, up to now the molecular interplay between cofilin and drebrin has not been elucidated. TIRF microscopy and solution experiments revealed that full length drebrin A or its actin binding core (Drb1-300) inhibits, but do not abolish cofilin-induced severing of actin filaments. Cosedimentation experiments showed that F-actin can be fully occupied with combination of these two proteins. The dependence of cofilin binding on fractional saturation of actin filaments with drebrin suggests direct competition between these two proteins for F-actin binding. This implies that cofilin and drebrin can either overcome or reverse the allosteric changes in F-actin induced by the competitor's binding. The ability of cofilin to displace drebrin from actin filaments is pH dependent and is facilitated at acidic pH (6.8). Pre-steady state kinetic experiments reveal that both binding and dissociation of drebrin to/from actin filaments is faster than that reported for cooperative binding of cofilin. We found, that drebrin displacement by cofilin is greatly inhibited when actin severing is abolished, which might be linked to the cooperativity of drebrin binding to actin filaments. Our results contribute to molecular understanding of the competitive interactions of drebrin and cofilin with actin filaments. PMID:25047716

  8. Targeting the actin cytoskeleton: selective antitumor action via trapping PKCɛ.

    PubMed

    Foerster, F; Braig, S; Moser, C; Kubisch, R; Busse, J; Wagner, E; Schmoeckel, E; Mayr, D; Schmitt, S; Huettel, S; Zischka, H; Mueller, R; Vollmar, A M

    2014-01-01

    Targeting the actin cytoskeleton (CSK) of cancer cells offers a valuable strategy in cancer therapy. There are a number of natural compounds that interfere with the actin CSK, but the mode of their cytotoxic action and, moreover, their tumor-specific mechanisms are quite elusive. We used the myxobacterial compound Chondramide as a tool to first elucidate the mechanisms of cytotoxicity of actin targeting in breast cancer cells (MCF7, MDA-MB-231). Chondramide inhibits cellular actin filament dynamics shown by a fluorescence-based analysis (fluorescence recovery after photobleaching (FRAP)) and leads to apoptosis characterized by phosphatidylserine exposure, release of cytochrome C from mitochondria and finally activation of caspases. Chondramide enhances the occurrence of mitochondrial permeability transition (MPT) by affecting known MPT modulators: Hexokinase II bound to the voltage-dependent anion channel (VDAC) translocated from the outer mitochondrial membrane to the cytosol and the proapoptotic protein Bad were recruited to the mitochondria. Importantly, protein kinase C-ɛ (PKCɛ), a prosurvival kinase possessing an actin-binding site and known to regulate the hexokinase/VDAC interaction as well as Bad phosphorylation was identified as the link between actin CSK and apoptosis induction. PKCɛ, which was found overexpressed in breast cancer cells, accumulated in actin bundles induced by Chondramide and lost its activity. Our second goal was to characterize the potential tumor-specific action of actin-binding agents. As the nontumor breast epithelial cell line MCF-10A in fact shows resistance to Chondramide-induced apoptosis and notably express low level of PKCɛ, we suggest that trapping PKCɛ via Chondramide-induced actin hyperpolymerization displays tumor cell specificity. Our work provides a link between targeting the ubiquitously occurring actin CSK and selective inhibition of pro-tumorigenic PKCɛ, thus setting the stage for actin-stabilizing agents as

  9. Targeting the actin cytoskeleton: selective antitumor action via trapping PKCɛ

    PubMed Central

    Foerster, F; Braig, S; Moser, C; Kubisch, R; Busse, J; Wagner, E; Schmoeckel, E; Mayr, D; Schmitt, S; Huettel, S; Zischka, H; Mueller, R; Vollmar, A M

    2014-01-01

    Targeting the actin cytoskeleton (CSK) of cancer cells offers a valuable strategy in cancer therapy. There are a number of natural compounds that interfere with the actin CSK, but the mode of their cytotoxic action and, moreover, their tumor-specific mechanisms are quite elusive. We used the myxobacterial compound Chondramide as a tool to first elucidate the mechanisms of cytotoxicity of actin targeting in breast cancer cells (MCF7, MDA-MB-231). Chondramide inhibits cellular actin filament dynamics shown by a fluorescence-based analysis (fluorescence recovery after photobleaching (FRAP)) and leads to apoptosis characterized by phosphatidylserine exposure, release of cytochrome C from mitochondria and finally activation of caspases. Chondramide enhances the occurrence of mitochondrial permeability transition (MPT) by affecting known MPT modulators: Hexokinase II bound to the voltage-dependent anion channel (VDAC) translocated from the outer mitochondrial membrane to the cytosol and the proapoptotic protein Bad were recruited to the mitochondria. Importantly, protein kinase C-ɛ (PKCɛ), a prosurvival kinase possessing an actin-binding site and known to regulate the hexokinase/VDAC interaction as well as Bad phosphorylation was identified as the link between actin CSK and apoptosis induction. PKCɛ, which was found overexpressed in breast cancer cells, accumulated in actin bundles induced by Chondramide and lost its activity. Our second goal was to characterize the potential tumor-specific action of actin-binding agents. As the nontumor breast epithelial cell line MCF-10A in fact shows resistance to Chondramide-induced apoptosis and notably express low level of PKCɛ, we suggest that trapping PKCɛ via Chondramide-induced actin hyperpolymerization displays tumor cell specificity. Our work provides a link between targeting the ubiquitously occurring actin CSK and selective inhibition of pro-tumorigenic PKCɛ, thus setting the stage for actin-stabilizing agents as

  10. Mechanics of composite actin networks: in vitro and cellular perspectives

    NASA Astrophysics Data System (ADS)

    Upadhyaya, Arpita

    2014-03-01

    Actin filaments and associated actin binding proteins play an essential role in governing the mechanical properties of eukaryotic cells. Even though cells have multiple actin binding proteins (ABPs) that exist simultaneously to maintain the structural and mechanical integrity of the cellular cytoskeleton, how these proteins work together to determine the properties of actin networks is not well understood. The ABP, palladin, is essential for the integrity of cell morphology and movement during development. Palladin coexists with alpha-actinin in stress fibers and focal adhesions and binds to both actin and alpha-actinin. To obtain insight into how mutually interacting actin crosslinking proteins modulate the properties of actin networks, we have characterized the micro-structure and mechanics of actin networks crosslinked with palladin and alpha-actinin. Our studies on composite networks of alpha-actinin/palladin/actin show that palladin and alpha-actinin synergistically determine network viscoelasticity. We have further examined the role of palladin in cellular force generation and mechanosensing. Traction force microscopy revealed that TAFs are sensitive to substrate stiffness as they generate larger forces on substrates of increased stiffness. Contrary to expectations, knocking down palladin increased the forces generated by cells, and also inhibited the ability to sense substrate stiffness for very stiff gels. This was accompanied by significant differences in the actin organization and adhesion dynamics of palladin knock down cells. Perturbation experiments also suggest altered myosin activity in palladin KD cells. Our results suggest that the actin crosslinkers such as palladin and myosin motors coordinate for optimal cell function and to prevent aberrant behavior as in cancer metastasis.

  11. Closed membrane shapes with attached BAR domains subject to external force of actin filaments.

    PubMed

    Mesarec, Luka; Góźdź, Wojciech; Iglič, Veronika Kralj; Kralj, Samo; Iglič, Aleš

    2016-05-01

    Membrane deformations induced by attached BAR superfamily domains could trigger or facilitate the growth of plasma membrane protrusions. The BAR domain family consists of BAR, F-BAR and I-BAR domains, each enforcing a different local curvature when attached to the membrane surface. Our theoretical study mainly focuses on the role of I-BAR in the membrane tubular deformations generated or stabilised by actin filaments. The influence of the area density of membrane attached BAR domains and their intrinsic curvature on the closed membrane shapes (vesicles) was investigated numerically. We derived an analytical approximative expression for the critical relative area density of BARs at which the membrane tubular protrusions on vesicles are most prominent. We have shown that the BARs with a higher intrinsic curvature induce thinner and longer cylindrical protrusions. The average orientation of the membrane attached BARs is altered when the vesicle shape is subjected to external force of growing actin rod-like structure inside a vesicle. The average orientation angle of membrane attached BARs may indicate whether the actin filaments are just stabilising the protrusion or generating it by stretching the vesicle. PMID:26854580

  12. A tridimensional view of the organization of actin filaments in the central nervous system by use of fluorescent photooxidation.

    PubMed

    Capani, Francisco; Saraceno, Ezequiel; Boti, Valeria Romina; Aon-Bertolino, Laura; Fernández, Juan Carlos; Gato, Fernándo; Kruse, Maria Sol; Krause, Maria Sol; Giraldez, Lisandro; Ellisman, Mark H; Coirini, Héctor

    2008-04-01

    Cellular and subcellular organization and distribution of actin filaments have been studied with various techniques. The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed the examination of the precise cellular and subcellular location of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are facilitated with this method for morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows the analysis of the three-dimensional location of F-actin with high-resolution light microscopy, three-d serial sections reconstructions, and electron tomography. The combination of selective staining and three-dimensional reconstructions provide a valuable tool for revealing aspects of the synaptic morphology that are not available when conventional electron microscopy is used. By applying this selective staining technique and three-dimensional imaging, we uncovered the structural organization of actin in the postsynaptic densities in physiological and pathological conditions. PMID:18669318

  13. Carbon Dioxide Flux Measurement Systems (CO2Flux) Handbook

    SciTech Connect

    Fischer, M

    2005-01-01

    The Southern Great Plains (SGP) carbon dioxide flux (CO2 flux) measurement systems provide half-hour average fluxes of CO2, H2O (latent heat), and sensible heat. The fluxes are obtained by the eddy covariance technique, which computes the flux as the mean product of the vertical wind component with CO2 and H2O densities, or estimated virtual temperature. A three-dimensional sonic anemometer is used to obtain the orthogonal wind components and the virtual (sonic) temperature. An infrared gas analyzer is used to obtain the CO2 and H2O densities. A separate sub-system also collects half-hour average measures of meteorological and soil variables from separate 4-m towers.

  14. Yeast studies reveal moonlighting functions of the ancient actin cytoskeleton.

    PubMed

    Sattlegger, Evelyn; Chernova, Tatiana A; Gogoi, Neeku M; Pillai, Indu V; Chernoff, Yury O; Munn, Alan L

    2014-08-01

    Classic functions of the actin cytoskeleton include control of cell size and shape and the internal organization of cells. These functions are manifest in cellular processes of fundamental importance throughout biology such as the generation of cell polarity, cell migration, cell adhesion, and cell division. However, studies in the unicellular model eukaryote Saccharomyces cerevisiae (Baker's yeast) are giving insights into other functions in which the actin cytoskeleton plays a critical role. These include endocytosis, control of protein translation, and determination of protein 3-dimensional shape (especially conversion of normal cellular proteins into prions). Here, we present a concise overview of these new "moonlighting" roles for the actin cytoskeleton and how some of these roles might lie at the heart of important molecular switches. This is an exciting time for researchers interested in the actin cytoskeleton. We show here how studies of actin are leading us into many new and exciting realms at the interface of genetics, biochemistry, and cell biology. While many of the pioneering studies have been conducted using yeast, the conservation of the actin cytoskeleton and its component proteins throughout eukaryotes suggests that these new roles for the actin cytoskeleton may not be restricted to yeast cells but rather may reflect new roles for the actin cytoskeleton of all eukaryotes. PMID:25138357

  15. Yeast studies reveal moonlighting functions of the ancient actin cytoskeleton

    PubMed Central

    Sattlegger, Evelyn; Chernova, Tatiana A.; Gogoi, Neeku M.; Pillai, Indu V.; Chernoff, Yury O.; Munn, Alan L.

    2014-01-01

    Classic functions of the actin cytoskeleton include control of cell size and shape and the internal organisation of cells. These functions are manifest in cellular processes of fundamental importance throughout biology such as the generation of cell polarity, cell migration, cell adhesion and cell division. However, studies in the unicellular model eukaryote Saccharomyces cerevisiae (Baker's yeast) are giving insights into other functions in which the actin cytoskeleton plays a critical role. These include endocytosis, control of protein translation and determination of protein 3-dimensional shape (especially conversion of normal cellular proteins into prions). Here we present a concise overview of these new "moonlighting" roles for the actin cytoskeleton and how some of these roles might lie at the heart of important molecular switches. This is an exciting time for researchers interested in the actin cytoskeleton. We show here how studies of actin are leading us into many new and exciting realms at the interface of genetics, biochemistry and cell biology. While many of the pioneering studies have been conducted using yeast, the conservation of the actin cytoskeleton and its component proteins throughout eukaryotes suggests that these new roles for the actin cytoskeleton may not be restricted to yeast cells but rather may reflect new roles for the actin cytoskeleton of all eukaryotes. PMID:25138357

  16. Deafness and espin-actin self-organization in stereocilia

    NASA Astrophysics Data System (ADS)

    Wong, Gerard C. L.

    2009-03-01

    Espins are F-actin-bundling proteins associated with large parallel actin bundles found in hair cell stereocilia in the ear, as well as brush border microvilli and Sertoli cell junctions. We examine actin bundle structures formed by different wild-type espin isoforms, fragments, and naturally-occurring human espin mutants linked to deafness and/or vestibular dysfunction. The espin-actin bundle structure consisted of a hexagonal arrangement of parallel actin filaments in a non-native twist state. We delineate the structural consequences caused by mutations in espin's actin-bundling module. For espin mutation with a severely damaged actin-bundling module, which are implicated in deafness in mice and humans, oriented nematic-like actin filament structures, which strongly impinges on bundle mechanical stiffness. Finally, we examine what makes espin different, via a comparative study of bundles formed by espin and those formed by fascin, a prototypical bundling protein found in functionally different regions of the cell, such as filopodia.

  17. Filament assembly by Spire: key residues and concerted actin binding.

    PubMed

    Rasson, Amy S; Bois, Justin S; Pham, Duy Stephen L; Yoo, Haneul; Quinlan, Margot E

    2015-02-27

    The most recently identified class of actin nucleators, WASp homology domain 2 (WH2) nucleators, use tandem repeats of monomeric actin-binding WH2 domains to facilitate actin nucleation. WH2 domains are involved in a wide variety of actin regulatory activities. Structurally, they are expected to clash with interprotomer contacts within the actin filament. Thus, the discovery of their role in nucleation was surprising. Here we use Drosophila Spire (Spir) as a model system to investigate both how tandem WH2 domains can nucleate actin and what differentiates nucleating WH2-containing proteins from their non-nucleating counterparts. We found that the third WH2 domain in Spir (Spir-C or SC) plays a unique role. In the context of a short nucleation construct (containing only two WH2 domains), placement of SC in the N-terminal position was required for the most potent nucleation. We found that the native organization of the WH2 domains with respect to each other is necessary for binding to actin with positive cooperativity. We identified two residues within SC that are critical for its activity. Using this information, we were able to convert a weak synthetic nucleator into one with activity equal to a native Spir construct. Lastly, we found evidence that SC binds actin filaments, in addition to monomers. PMID:25234086

  18. Filament Assembly by Spire: Key Residues and Concerted Actin Binding

    PubMed Central

    Rasson, Amy S.; Bois, Justin S.; Pham, Duy Stephen L.; Yoo, Haneul; Quinlan, Margot E.

    2014-01-01

    The most recently identified class of actin nucleators, WASp Homology domain 2 (WH2) – nucleators, use tandem repeats of monomeric actin-binding WH2 domains to facilitate actin nucleation. WH2 domains are involved in a wide variety of actin regulatory activities. Structurally, they are expected to clash with interprotomer contacts within the actin filament. Thus, the discovery of their role in nucleation was surprising. Here we use Drosophila Spire (Spir) as a model system to investigate both how tandem WH2 domains can nucleate actin and what differentiates nucleating WH2-containing proteins from their non-nucleating counterparts. We found that the third WH2 domain in Spir (Spir-C or Sc), plays a unique role. In the context of a short nucleation construct (containing only two WH2 domains), placement of Sc in the N-terminal position was required for the most potent nucleation. We found that the native organization of the WH2 domains with respect to each other is necessary for binding to actin with positive cooperativity. We identified two residues within Sc that are critical for its activity. Using this information we were able to convert a weak synthetic nucleator into one with activity equal to a native Spir construct. Lastly, we found evidence that Sc binds actin filaments, in addition to monomers. PMID:25234086

  19. Actin-based spindle positioning: new insights from female gametes.

    PubMed

    Almonacid, Maria; Terret, Marie-Émilie; Verlhac, Marie-Hélène

    2014-02-01

    Asymmetric divisions are essential in metazoan development, where they promote the emergence of cell lineages. The mitotic spindle has astral microtubules that contact the cortex, which act as a sensor of cell geometry and as an integrator to orient cell division. Recent advances in live imaging revealed novel pools and roles of F-actin in somatic cells and in oocytes. In somatic cells, cytoplasmic F-actin is involved in spindle architecture and positioning. In starfish and mouse oocytes, newly discovered meshes of F-actin control chromosome gathering and spindle positioning. Because oocytes lack centrosomes and astral microtubules, F-actin networks are key players in the positioning of spindles by transmitting forces over long distances. Oocytes also achieve highly asymmetric divisions, and thus are excellent models to study the roles of these newly discovered F-actin networks in spindle positioning. Moreover, recent studies in mammalian oocytes provide a further understanding of the organisation of F-actin networks and their biophysical properties. In this Commentary, we present examples of the role of F-actin in spindle positioning and asymmetric divisions, with an emphasis on the most up-to-date studies from mammalian oocytes. We also address specific technical issues in the field, namely live imaging of F-actin networks and stress the need for interdisciplinary approaches. PMID:24413163

  20. Actin Interacts with Dengue Virus 2 and 4 Envelope Proteins

    PubMed Central

    Jitoboam, Kunlakanya; Phaonakrop, Narumon; Libsittikul, Sirikwan; Thepparit, Chutima; Roytrakul, Sittiruk; Smith, Duncan R.

    2016-01-01

    Dengue virus (DENV) remains a significant public health problem in many tropical and sub-tropical countries worldwide. The DENV envelope (E) protein is the major antigenic determinant and the protein that mediates receptor binding and endosomal fusion. In contrast to some other DENV proteins, relatively few cellular interacting proteins have been identified. To address this issue a co-immuoprecipitation strategy was employed. The predominant co-immunoprecipitating proteins identified were actin and actin related proteins, however the results suggested that actin was the only bona fide interacting partner. Actin was shown to interact with the E protein of DENV 2 and 4, and the interaction between actin and DENV E protein was shown to occur in a truncated DENV consisting of only domains I and II. Actin was shown to decrease during infection, but this was not associated with a decrease in gene transcription. Actin-related proteins also showed a decrease in expression during infection that was not transcriptionally regulated. Cytoskeletal reorganization was not observed during infection, suggesting that the interaction between actin and E protein has a cell type specific component. PMID:27010925

  1. Building an artificial actin cortex on microscopic pillar arrays.

    PubMed

    Ayadi, R; Roos, W H

    2015-01-01

    Eukaryotic cells obtain their morphology and mechanical strength from the cytoskeleton and in particular from the cross-linked actin network that branches throughout the whole cell. This actin cortex lies like a quasi-two-dimensional (2D) biopolymer network just below the cell membrane, to which it is attached. In the quest for building an artificial cell, one needs to make a biomimetic model of the actin cortex and combine this in a bottom-up approach with other "synthetic" components. Here, we describe a reconstitution method for such an artificial actin cortex, which is freely suspended on top of a regular array of pillars. By this immobilization method, the actin network is only attached to a surface at discrete points and can fluctuate freely in between. By discussing the method to make the micropillars and the way to reconstitute a quasi-2D actin network on top, we show how one can study an isolated, reconstituted part of a cell. This allows the study of fundamental interaction mechanisms of actin networks, providing handles to design a functional actin cortex in an artificial cell. PMID:25997345

  2. Bending Flexibility of Actin Filaments during Motor-Induced Sliding

    PubMed Central

    Vikhorev, Petr G.; Vikhoreva, Natalia N.; Månsson, Alf

    2008-01-01

    Muscle contraction and other forms of cell motility occur as a result of cyclic interactions between myosin molecules and actin filaments. Force generation is generally attributed to ATP-driven structural changes in myosin, whereas a passive role is ascribed to actin. However, some results challenge this view, predicting structural changes in actin during motor activity, e.g., when the actin filaments slide on a myosin-coated surface in vitro. Here, we analyzed statistical properties of the sliding filament paths, allowing us to detect changes of this type. It is interesting to note that evidence for substantial structural changes that led to increased bending flexibility of the filaments was found in phalloidin-stabilized, but not in phalloidin-free, actin filaments. The results are in accordance with the idea that a high-flexibility structural state of actin is a prerequisite for force production, but not the idea that a low-to-high flexibility transition of the actin filament should be an important component of the force-generating step per se. Finally, our data challenge the general view that phalloidin-stabilized filaments behave as native actin filaments in their interaction with myosin. This has important implications, since phalloidin stabilization is a routine procedure in most studies of actomyosin function. PMID:18835897

  3. Toll-like receptor ligands sensitize B-cell receptor signalling by reducing actin-dependent spatial confinement of the receptor

    PubMed Central

    Freeman, Spencer A.; Jaumouillé, Valentin; Choi, Kate; Hsu, Brian E.; Wong, Harikesh S.; Abraham, Libin; Graves, Marcia L.; Coombs, Daniel; Roskelley, Calvin D.; Das, Raibatak; Grinstein, Sergio; Gold, Michael R.

    2015-01-01

    Integrating signals from multiple receptors allows cells to interpret the physiological context in which a signal is received. Here we describe a mechanism for receptor crosstalk in which receptor-induced increases in actin dynamics lower the threshold for signalling by another receptor. We show that the Toll-like receptor ligands lipopolysaccharide and CpG DNA, which are conserved microbial molecules, enhance signalling by the B-cell antigen receptor (BCR) by activating the actin-severing protein cofilin. Single-particle tracking reveals that increased severing of actin filaments reduces the spatial confinement of the BCR within the plasma membrane and increases BCR mobility. This allows more frequent collisions between BCRs and greater signalling in response to low densities of membrane-bound antigen. These findings implicate actin dynamics as a means of tuning receptor signalling and as a mechanism by which B cells distinguish inert antigens from those that are accompanied by indicators of microbial infection. PMID:25644899

  4. G-actin guides p53 nuclear transport: potential contribution of monomeric actin in altered localization of mutant p53

    PubMed Central

    Saha, Taniya; Guha, Deblina; Manna, Argha; Panda, Abir Kumar; Bhat, Jyotsna; Chatterjee, Subhrangsu; Sa, Gaurisankar

    2016-01-01

    p53 preserves genomic integrity by restricting anomaly at the gene level. Till date, limited information is available for cytosol to nuclear shuttling of p53; except microtubule-based trafficking route, which utilizes minus-end directed motor dynein. The present study suggests that monomeric actin (G-actin) guides p53 traffic towards the nucleus. Histidine-tag pull-down assay using purified p53(1–393)-His and G-actin confirms direct physical association between p53 and monomeric G-actin. Co-immunoprecipitation data supports the same. Confocal imaging explores intense perinuclear colocalization between p53 and G-actin. To address atomistic details of the complex, constraint-based docked model of p53:G-actin complex was generated based on crystal structures. MD simulation reveals that p53 DNA-binding domain arrests very well the G-actin protein. Docking benchmark studies have been carried out for a known crystal structure, 1YCS (complex between p53DBD and BP2), which validates the docking protocol we adopted. Co-immunoprecipitation study using “hot-spot” p53 mutants suggested reduced G-actin association with cancer-associated p53 conformational mutants (R175