The Regulation of Filamentous Growth in Yeast

PubMed Central

Cullen, Paul J.; Sprague, George F.

2012-01-01

Filamentous growth is a nutrient-regulated growth response that occurs in many fungal species. In pathogens, filamentous growth is critical for host–cell attachment, invasion into tissues, and virulence. The budding yeast Saccharomyces cerevisiae undergoes filamentous growth, which provides a genetically tractable system to study the molecular basis of the response. Filamentous growth is regulated by evolutionarily conserved signaling pathways. One of these pathways is a mitogen activated protein kinase (MAPK) pathway. A remarkable feature of the filamentous growth MAPK pathway is that it is composed of factors that also function in other pathways. An intriguing challenge therefore has been to understand how pathways that share components establish and maintain their identity. Other canonical signaling pathways—rat sarcoma/protein kinase A (RAS/PKA), sucrose nonfermentable (SNF), and target of rapamycin (TOR)—also regulate filamentous growth, which raises the question of how signals from multiple pathways become integrated into a coordinated response. Together, these pathways regulate cell differentiation to the filamentous type, which is characterized by changes in cell adhesion, cell polarity, and cell shape. How these changes are accomplished is also discussed. High-throughput genomics approaches have recently uncovered new connections to filamentous growth regulation. These connections suggest that filamentous growth is a more complex and globally regulated behavior than is currently appreciated, which may help to pave the way for future investigations into this eukaryotic cell differentiation behavior. PMID:22219507

The vacuole system is a significant intracellular pathway for longitudinal solute transport in basidiomycete fungi.

PubMed

Darrah, P R; Tlalka, M; Ashford, A; Watkinson, S C; Fricker, M D

2006-07-01

Mycelial fungi have a growth form which is unique among multicellular organisms. The data presented here suggest that they have developed a unique solution to internal solute translocation involving a complex, extended vacuole. In all filamentous fungi examined, this extended vacuole forms an interconnected network, dynamically linked by tubules, which has been hypothesized to act as an internal distribution system. We have tested this hypothesis directly by quantifying solute movement within the organelle by photobleaching a fluorescent vacuolar marker. Predictive simulation models were then used to determine the transport characteristics over extended length scales. This modeling showed that the vacuolar organelle forms a functionally important, bidirectional diffusive transport pathway over distances of millimeters to centimeters. Flux through the pathway is regulated by the dynamic tubular connections involving homotypic fusion and fission. There is also a strongly predicted interaction among vacuolar organization, predicted diffusion transport distances, and the architecture of the branching colony margin.

pH measurement of tubular vacuoles of an arbuscular mycorrhizal fungus, Gigaspora margarita.

PubMed

Funamoto, Rintaro; Saito, Katsuharu; Oyaizu, Hiroshi; Aono, Toshihiro; Saito, Masanori

2015-01-01

Arbuscular mycorrhizal fungi play an important role in phosphate supply to the host plants. The fungal hyphae contain tubular vacuoles where phosphate compounds such as polyphosphate are accumulated. Despite their importance for the phosphate storage, little is known about the physiological properties of the tubular vacuoles in arbuscular mycorrhizal fungi. As an indicator of the physiological state in vacuoles, we measured pH of tubular vacuoles in living hyphae of arbuscular mycorrhizal fungus Gigaspora margarita using ratio image analysis with pH-dependent fluorescent probe, 6-carboxyfluorescein. Fluorescent images of the fine tubular vacuoles were obtained using a laser scanning confocal microscope, which enabled calculation of vacuolar pH with high spatial resolution. The tubular vacuoles showed mean pH of 5.6 and a pH range of 5.1-6.3. These results suggest that the tubular vacuoles of arbuscular mycorrhizal fungi have a mildly acidic pH just like vacuoles of other fungal species including yeast and ectomycorrhizal fungi.

Co-occurrence of tannin and tannin-less vacuoles in sensitive plants.

PubMed

Fleurat-Lessard, Pierrette; Béré, Emile; Lallemand, Magali; Dédaldéchamp, Fabienne; Roblin, Gabriel

2016-05-01

Vacuoles of different types frequently coexist in the same plant cell, but the duality of the tannin/tannin-less vacuoles observed in Mimosa pudica L. is rare. In this plant, which is characterized by highly motile leaves, the development and original features of the double vacuolar compartment were detailed in primary pulvini from the young to the mature leaf stage. In young pulvini, the differentiation of tannin vacuoles first occurred in the epidermis and progressively spread toward the inner cortex. In motor cells of nonmotile pulvini, tannin deposits first lined the membranes of small vacuole profiles and then formed opaque clusters that joined together to form a large tannin vacuole (TV), the proportion of which in the cell was approximately 45%. At this stage, transparent vacuole profiles were rare and small, but as the parenchyma cells enlarged, these profiles coalesced to form a transparent vacuole with a convexity toward the larger-sized tannin vacuole. When leaf motility began to occur, the two vacuole types reached the same relative proportion (approximately 30%). Finally, in mature cells displaying maximum motility, the large transparent colloidal vacuole (CV) showed a relative proportion increasing to approximately 50%. At this stage, the proportion of the tannin vacuole, occurring in the vicinity of the nucleus, decreased to approximately 10%. The presence of the condensed type of tannins (proanthocyanidins) was proven by detecting their fluorescence under UV light and by specific chemical staining. This dual vacuolar profile was also observed in nonmotile parts of M. pudica (e.g., the petiole and the stem). Additional observations of leaflet pulvini showing more or less rapid movements showed that this double vacuolar structure was present in certain plants (Mimosa spegazzinii and Desmodium gyrans), but absent in others (Albizzia julibrissin, Biophytum sensitivum, and Cassia fasciculata). Taken together, these observations strongly suggest that a

Colloidal transport by active filaments

NASA Astrophysics Data System (ADS)

Manna, Raj Kumar; Kumar, P. B. Sunil; Adhikari, R.

2017-01-01

Enhanced colloidal transport beyond the limit imposed by diffusion is usually achieved through external fields. Here, we demonstrate the ballistic transport of a colloidal sphere using internal sources of energy provided by an attached active filament. The latter is modeled as a chain of chemo-mechanically active beads connected by potentials that enforce semi-flexibility and self-avoidance. The fluid flow produced by the active beads and the forces they mediate are explicitly taken into account in the overdamped equations of motion describing the colloid-filament assembly. The speed and efficiency of transport depend on the dynamical conformational states of the filament. We characterize these states using filament writhe as an order parameter and identify ones yielding maxima in speed and efficiency of transport. The transport mechanism reported here has a remarkable resemblance to the flagellar propulsion of microorganisms which suggests its utility in biomimetic systems.

Scroll wave filaments self-wrap around unexcitable heterogeneities.

PubMed

Jiménez, Zulma A; Steinbock, Oliver

2012-09-01

Scroll waves are three-dimensional excitation vortices rotating around one-dimensional phase singularities called filaments. In experiments with a chemical reaction-diffusion system and in numerical simulations, we study the pinning of closed filament loops to inert cylindrical heterogeneities. We show that the filament wraps itself around the heterogeneity and thus avoids contraction and annihilation. This entwining steadily increases the total length of the pinned filament and reshapes the entire rotation backbone of the vortex. Self-pinning is fastest for thin cylinders with radii not much larger than the core of the unpinned rotor. The process ends when the filament is attached to the entire length of the cylinder. The possible importance of self-pinning in cardiac systems is discussed.

Self-assembly of actin monomers into long filaments: Brownian dynamics simulations

NASA Astrophysics Data System (ADS)

Guo, Kunkun; Shillcock, Julian; Lipowsky, Reinhard

2009-07-01

Brownian dynamics simulations are used to study the dynamical process of self-assembly of actin monomers into long filaments containing up to 1000 actin protomers. In order to overcome the large separation of time scales between the diffusive motion of the free monomers and the relatively slow attachment and detachment processes at the two ends of the filaments, we introduce a novel rescaling procedure by which we speed all dynamical processes related to actin polymerization and depolymerization up by the same factor. In general, the actin protomers within a filament can attain three different states corresponding to a bound adenosine triphosphate (ATP), adenosine diphosphate with inorganic phosphate (ADP/P), and ADP molecule. The simplest situation that has been studied experimentally is provided by the polymerization of ADP-actin, for which all protomers are identical. This case is used to unravel certain relations between the filament's physical properties and the model parameters such as the attachment rate constant and the size of the capture zone, the detachment rate and the probability of the detached event, as well as the growth rate and waiting times between two successive attachment/detachment events. When a single filament is allowed to grow in a bath of constant concentration of free ADP-actin monomers, its growth rate increases linearly with the free monomer concentration in quantitative agreement with in vitro experiments. The results also show that the waiting time is governed by exponential distributions and that the two ends of a filament undergo biased random walks. The filament length fluctuations are described by a length diffusion constant that is found to attain a constant value at low ADP-actin concentration and to increase linearly with this concentration. It is straightforward to apply our simulation code to more complex processes such as polymerization of ATP-actin coupled to ATP hydrolysis, force generation by filaments, formation of

Intermicrotubular actin filaments in the transalar cytoskeletal arrays of Drosophila.

PubMed

Mogensen, M M; Tucker, J B

1988-11-01

Rabbit muscle myosin subfragment S1 decorates 6 nm diameter filaments in Drosophila wing epidermal cells in the arrowhead fashion characteristic of the binding of subfragment S1 to actin filaments. The filaments in question are concentrated between microtubules that are mostly composed of 15 protofilaments and form cell surface-associated transcellular bundles. There are indications that the majority of the actin filaments have the same polarity and that, like the microtubules, they may elongate from sites at the apical surfaces of the cells. The bundles of F actin and microtubules occur in dorsal and ventral epidermal cell layers of a wing blade. They are joined in dorso-ventral pairs by attachment desmosomes. These transalar cytoskeletal arrays may provide an example of a situation where actin filaments operate as stiffeners rather than active generators of force in conjunction with myosin. The arrays probably function as noncontractile pillars to maintain basal cell extensions and keep haemocoelic spaces open in the highly folded and expanding wing blades of late pupae.

Molecular gas in high-mass filament WB673

NASA Astrophysics Data System (ADS)

Kirsanova, Maria S.; Salii, Svetlana V.; Sobolev, Andrej M.; Olofsson, Anders Olof Henrik; Ladeyschikov, Dmitry A.; Thomasson, Magnus

2017-12-01

We studied the distribution of dense gas in a filamentary molecular cloud containing several dense clumps. The center of the filament is given by the dense clump WB673. The clumps are high-mass and intermediate-mass starforming regions. We observed CS (2-1), 13CO (1-0), C18O(1-0), and methanol lines at 96 GHz toward WB673 with the Onsala Space Observatory 20-m telescope. We found CS (2-1) emission in the inter-clump medium so the clumps are physically connected and the whole cloud is indeed a filament. Its total mass is 104 M⊙ and mass-to-length ratio is 360M⊙ pc-1 from 13CO (1-0) data. Mass-to-length ratio for the dense gas is 3.4 - 34M⊙ pc-1 from CS (2-1) data. The PV-diagram of the filament is V-shaped. We estimated physical conditions in the molecular gas using methanol lines. Location of the filament on the sky between extended shells suggests that it could be a good example to test theoretical models of formation of the filaments via multiple compression of interstellar gas by supersonic waves.

Cell Vacuolation Caused by Vibrio cholerae Hemolysin

PubMed Central

Figueroa-Arredondo, Paula; Heuser, John E.; Akopyants, Natalia S.; Morisaki, J. Hiroshi; Giono-Cerezo, Silvia; Enríquez-Rincón, Fernando; Berg, Douglas E.

2001-01-01

Non-O1 strains of Vibrio cholerae implicated in gastroenteritis and diarrhea generally lack virulence determinants such as cholera toxin that are characteristic of epidemic strains; the factors that contribute to their virulence are not understood. Here we report that at least one-third of diarrhea-associated nonepidemic V. cholerae strains from Mexico cause vacuolation of cultured Vero cells. Detailed analyses indicated that this vacuolation was related to that caused by aerolysin, a pore-forming toxin of Aeromonas; it involved primarily the endoplasmic reticulum at early times (∼1 to 4 h after exposure), and resulted in formation of large, acidic, endosome-like multivesicular vacuoles (probably autophagosomes) only at late times (∼16 h). In contrast to vacuolation caused by Helicobacter pylori VacA protein, that induced by V. cholerae was exacerbated by agents that block vacuolar proton pumping but not by endosome-targeted weak bases. It caused centripetal redistribution of endosomes, reflecting cytoplasmic alkalinization. The gene for V. cholerae vacuolating activity was cloned and was found to correspond to hlyA, the structural gene for hemolysin. HlyA protein is a pore-forming toxin that causes ion leakage and, ultimately, eukaryotic cell lysis. Thus, a distinct form of cell vacuolation precedes cytolysis at low doses of hemolysin. We propose that this vacuolation, in itself, contributes to the virulence of V. cholerae strains, perhaps by perturbing intracellular membrane trafficking or ion exchange in target cells and thereby affecting local intestinal inflammatory or other defense responses. PMID:11179335

Cell vacuolation caused by Vibrio cholerae hemolysin.

PubMed

Figueroa-Arredondo, P; Heuser, J E; Akopyants, N S; Morisaki, J H; Giono-Cerezo, S; Enríquez-Rincón, F; Berg, D E

2001-03-01

Non-O1 strains of Vibrio cholerae implicated in gastroenteritis and diarrhea generally lack virulence determinants such as cholera toxin that are characteristic of epidemic strains; the factors that contribute to their virulence are not understood. Here we report that at least one-third of diarrhea-associated nonepidemic V. cholerae strains from Mexico cause vacuolation of cultured Vero cells. Detailed analyses indicated that this vacuolation was related to that caused by aerolysin, a pore-forming toxin of Aeromonas; it involved primarily the endoplasmic reticulum at early times (approximately 1 to 4 h after exposure), and resulted in formation of large, acidic, endosome-like multivesicular vacuoles (probably autophagosomes) only at late times (approximately 16 h). In contrast to vacuolation caused by Helicobacter pylori VacA protein, that induced by V. cholerae was exacerbated by agents that block vacuolar proton pumping but not by endosome-targeted weak bases. It caused centripetal redistribution of endosomes, reflecting cytoplasmic alkalinization. The gene for V. cholerae vacuolating activity was cloned and was found to correspond to hlyA, the structural gene for hemolysin. HlyA protein is a pore-forming toxin that causes ion leakage and, ultimately, eukaryotic cell lysis. Thus, a distinct form of cell vacuolation precedes cytolysis at low doses of hemolysin. We propose that this vacuolation, in itself, contributes to the virulence of V. cholerae strains, perhaps by perturbing intracellular membrane trafficking or ion exchange in target cells and thereby affecting local intestinal inflammatory or other defense responses.

The Role of Magnetic Fields in High-Mass Star-Forming Filaments

NASA Astrophysics Data System (ADS)

Stephens, Ian

2015-10-01

Filaments are ubiquitous in the star formation process. Planck has revealed that magnetic fields are perpendicular to the densest filaments, which are the birthplace of high-mass stars, suggesting that fields help funnel gas into the filaments. However, the resolved field morphologies and strengths in the dense filaments are unknown. We propose HAWC+ 53 and 214 um polarimetric observations toward two filaments, the Snake (G11.1) and G18.6, to unveil the field morphology. Such observations will probe the filament field morphology at the subarcminute scale over the largest spatial extent to date: 25 and 9 pc respectively. We expect to have over 400 independent beams worth of detections. From the field morphology, we will test the hub-filament theory and investigate how the magnetic field strength and morphology changes with evolution and size-scale.

The Role of Magnetic Fields in High-Mass Star-Forming Filaments

NASA Astrophysics Data System (ADS)

Stephens, Ian

Filaments are ubiquitous in the star formation process. Planck has revealed that magnetic fields are perpendicular to the densest filaments, which are the birthplace of high-mass stars, suggesting that fields help funnel gas into the filaments. However, the resolved field morphologies and strengths in the dense filaments are unknown. We propose HAWC+ 53 and 214 um polarimetric observations toward two filaments, the Snake (G11.1) and G18.6, to unveil the field morphology. Such observations will probe the filament field morphology at the subarcminute scale over the largest spatial extent to date: 25 and 9 pc respectively. We expect to have over 400 independent beams worth of detections. From the field morphology, we will test the hub-filament theory and investigate how the magnetic field strength and morphology changes with evolution and size-scale.

Cellular vacuolation induced by Clostridium perfringens epsilon-toxin.

PubMed

Nagahama, Masahiro; Itohayashi, Yukari; Hara, Hideki; Higashihara, Masahiro; Fukatani, Yusuke; Takagishi, Teruhisa; Oda, Masataka; Kobayashi, Keiko; Nakagawa, Ichiro; Sakurai, Jun

2011-09-01

The epsilon-toxin of Clostridium perfringens forms a heptamer in the membranes of Madin-Darby canine kidney cells, leading to cell death. Here, we report that it caused the vacuolation of Madin-Darby canine kidney cells. The toxin induced vacuolation in a dose-dependent and time-dependent manner. The monomer of the toxin formed oligomers on lipid rafts in membranes of the cells. Methyl-β-cyclodextrin and poly(ethylene glycol) 4000 inhibited the vacuolation. Epsilon-toxin was internalized into the cells. Confocal microscopy revealed that the internalized toxin was transported from early endosomes (early endosome antigen 1 staining) to late endosomes and lysosomes (lysosomal-associated membrane protein 2 staining) and then distributed to the membranes of vacuoles. Furthermore, the vacuolation was inhibited by bafilomycin A1, a V-type ATPase inhibitor, and colchicine and nocodazole, microtubule-depolymerizing agents. The early endosomal marker green fluorescent protein-Rab5 and early endosome antigen 1 did not localize to vacuolar membranes. In contrast, the vacuolar membranes were specifically stained by the late endosomal and lysosomal marker green fluorescent protein-Rab7 and lysosomal-associated membrane protein 2. The vacuoles in the toxin-treated cells were stained with LysoTracker Red DND-99, a marker for late endosomes and lysosomes. A dominant negative mutant of Rab7 prevented the vacuolization, whereas a mutant form of Rab5 was less effective. These results demonstrate, for the first time, that: (a) oligomers of epsilon-toxin formed in lipid rafts are endocytosed; and (b) the vacuoles originating from late endosomes and lysosomes are formed by an oligomer of epsilon-toxin. © 2011 The Authors Journal compilation © 2011 FEBS.

Notochord vacuoles are lysosome-related organelles that function in axis and spine morphogenesis.

PubMed

Ellis, Kathryn; Bagwell, Jennifer; Bagnat, Michel

2013-03-04

The notochord plays critical structural and signaling roles during vertebrate development. At the center of the vertebrate notochord is a large fluid-filled organelle, the notochord vacuole. Although these highly conserved intracellular structures have been described for decades, little is known about the molecular mechanisms involved in their biogenesis and maintenance. Here we show that zebrafish notochord vacuoles are specialized lysosome-related organelles whose formation and maintenance requires late endosomal trafficking regulated by the vacuole-specific Rab32a and H(+)-ATPase-dependent acidification. We establish that notochord vacuoles are required for body axis elongation during embryonic development and identify a novel role in spine morphogenesis. Thus, the vertebrate notochord plays important structural roles beyond early development.

Notochord vacuoles are lysosome-related organelles that function in axis and spine morphogenesis

PubMed Central

Ellis, Kathryn; Bagwell, Jennifer

2013-01-01

The notochord plays critical structural and signaling roles during vertebrate development. At the center of the vertebrate notochord is a large fluid-filled organelle, the notochord vacuole. Although these highly conserved intracellular structures have been described for decades, little is known about the molecular mechanisms involved in their biogenesis and maintenance. Here we show that zebrafish notochord vacuoles are specialized lysosome-related organelles whose formation and maintenance requires late endosomal trafficking regulated by the vacuole-specific Rab32a and H+-ATPase–dependent acidification. We establish that notochord vacuoles are required for body axis elongation during embryonic development and identify a novel role in spine morphogenesis. Thus, the vertebrate notochord plays important structural roles beyond early development. PMID:23460678

Paired Helical Filaments of Inclusion-Body Myositis Muscle Contain RNA and Survival Motor Neuron Protein

PubMed Central

Broccolini, Aldobrando; Engel, W. King; Alvarez, Renate B.; Askanas, Valerie

2000-01-01

Sporadic inclusion-body myositis (s-IBM) is the most common progressive muscle disease of older persons. Pathologically, the muscle biopsy manifests various degrees of inflammation and specific vacuolar degeneration of muscle fibers characterized by paired helical filaments (PHFs) composed of phosphorylated tau. IBM vacuolated fibers also contain accumulations of several other Alzheimer-characteristic proteins. Molecular mechanisms leading to formation of the PHFs and accumulations of proteins in IBM muscle are not known. We report that the abnormal muscle fibers of IBM contained (i) acridine-orange-positive RNA inclusions that colocalized with the immunoreactivity of phosphorylated tau and (ii) survival motor neuron protein immunoreactive inclusions, which by immuno-electron microscopy were confined to paired helical filaments. This study demonstrates two novel components of the IBM paired helical filaments, which may lead to better understanding of their pathogenesis. PMID:10751338

Paired helical filaments of inclusion-body myositis muscle contain RNA and survival motor neuron protein.

PubMed

Broccolini, A; Engel, W K; Alvarez, R B; Askanas, V

2000-04-01

Sporadic inclusion-body myositis (s-IBM) is the most common progressive muscle disease of older persons. Pathologically, the muscle biopsy manifests various degrees of inflammation and specific vacuolar degeneration of muscle fibers characterized by paired helical filaments (PHFs) composed of phosphorylated tau. IBM vacuolated fibers also contain accumulations of several other Alzheimer-characteristic proteins. Molecular mechanisms leading to formation of the PHFs and accumulations of proteins in IBM muscle are not known. We report that the abnormal muscle fibers of IBM contained (i) acridine-orange-positive RNA inclusions that colocalized with the immunoreactivity of phosphorylated tau and (ii) survival motor neuron protein immunoreactive inclusions, which by immuno-electron microscopy were confined to paired helical filaments. This study demonstrates two novel components of the IBM paired helical filaments, which may lead to better understanding of their pathogenesis.

Giant quiescent solar filament observed with high-resolution spectroscopy

NASA Astrophysics Data System (ADS)

Kuckein, C.; Verma, M.; Denker, C.

2016-05-01

Aims: An extremely large filament was studied in various layers of the solar atmosphere. The inferred physical parameters and the morphological aspects are compared with smaller quiescent filaments. Methods: A giant quiet-Sun filament was observed with the high-resolution Echelle spectrograph at the Vacuum Tower Telescope at Observatorio del Teide, Tenerife, Spain, on 2011 November 15. A mosaic of spectra (ten maps of 100″ × 182″) was recorded simultaneously in the chromospheric absorption lines Hα and Na I D2. Physical parameters of the filament plasma were derived using cloud model (CM) inversions and line core fits. The spectra were complemented with full-disk filtergrams (He I λ10830 Å, Hα, and Ca II K) of the Chromospheric Telescope (ChroTel) and full-disk magnetograms of the Helioseismic and Magnetic Imager (HMI). Results: The filament had extremely large linear dimensions (~817 arcsec), which corresponds to about 658 Mm along a great circle on the solar surface. A total amount of 175119 Hα contrast profiles were inverted using the CM approach. The inferred mean line-of-sight (LOS) velocity, Doppler width, and source function were similar to previous works of smaller quiescent filaments. However, the derived optical thickness was higher. LOS velocity trends inferred from the Hα line core fits were in accord but weaker than those obtained with CM inversions. Signatures of counter-streaming flows were detected in the filament. The largest brightening conglomerates in the line core of Na I D2 coincided well with small-scale magnetic fields as seen by HMI. Mixed magnetic polarities were detected close to the ends of barbs. The computation of photospheric horizontal flows based on HMI magnetograms revealed flow kernels with a size of 5-8 Mm and velocities of 0.30-0.45 km s-1 at the ends of the filament. Conclusions: The physical properties of extremely large filaments are similar to their smaller counterparts, except for the optical thickness, which in

The Toxoplasma Parasitophorous Vacuole: An Evolving Host-Parasite Frontier.

PubMed

Clough, Barbara; Frickel, Eva-Maria

2017-06-01

The parasitophorous vacuole is a unique replicative niche for apicomplexan parasites, including Toxoplasma gondii. Derived from host plasma membrane, the vacuole is rendered nonfusogenic with the host endolysosomal system. Toxoplasma secretes numerous proteins to modify the forming vacuole, enable nutrient uptake, and set up mechanisms of host subversion. Here we describe the pathways of host-parasite interaction at the parasitophorous vacuole employed by Toxoplasma and host, leading to the intricate balance of host defence versus parasite survival. Copyright © 2017 Elsevier Ltd. All rights reserved.

Filament wound data base development, revision 1, appendix A

NASA Technical Reports Server (NTRS)

Sharp, R. Scott; Braddock, William F.

1985-01-01

Data are presented in tabular form for the High Performance Nozzle Increments, Filament Wound Case (FWC) Systems Tunnel Increments, Steel Case Systems Tunnel Increments, FWC Stiffener Rings Increments, Steel Case Stiffener Rings Increments, FWC External Tank (ET) Attach Ring Increments, Steel Case ET Attach Ring Increments, and Data Tape 8. The High Performance Nozzle are also presented in graphical form. The tabular data consist of six-component force and moment coefficients as they vary with angle of attack at a specific Mach number and roll angle. The six coefficients are normal force, pitching moment, side force, yawing moment, axial force, and rolling moment. The graphical data for the High Performance Nozzle Increments consist of a plot of a coefficient increment as a function of angle of attack at a specific Mach number and at a roll angle of 0 deg.

High-resolution Observations of Sympathetic Filament Eruptions by NVST

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Shangwei; Su, Yingna; Zhou, Tuanhui

We investigate two sympathetic filament eruptions observed by the New Vacuum Solar Telescope on 2015 October 15. The full picture of the eruptions is obtained from the corresponding Solar Dynamics Observatory ( SDO )/Atmospheric Imaging Assembly (AIA) observations. The two filaments start from active region NOAA 12434 in the north and end in one large quiescent filament channel in the south. The left filament erupts first, followed by the right filament eruption about 10 minutes later. Clear twist structure and rotating motion are observed in both filaments during the eruption. Both eruptions failed, since the filaments first rise up, thenmore » flow toward the south and merge into the southern large quiescent filament. We also observe repeated activations of mini filaments below the right filament after its eruption. Using magnetic field models constructed based on SDO /HMI magnetograms via the flux rope insertion method, we find that the left filament eruption is likely to be triggered by kink instability, while the weakening of overlying magnetic fields due to magnetic reconnection at an X-point between the two filament systems might play an important role in the onset of the right filament eruption.« less

Heat-induced formation of myosin oligomer-soluble filament complex in high-salt solution.

PubMed

Shimada, Masato; Takai, Eisuke; Ejima, Daisuke; Arakawa, Tsutomu; Shiraki, Kentaro

2015-02-01

Heat-induced aggregation of myosin into an elastic gel plays an important role in the water-holding capacity and texture of meat products. Here, we investigated thermal aggregation of porcine myosin in high-salt solution over a wide temperature range by dynamic light scattering experiments. The myosin samples were readily dissolved in 1.0 M NaCl at 25 °C followed by dilution into various salt concentrations. The diluted solutions consistently contained both myosin monomers and soluble filaments. The filament size decreased with increasing salt concentration and temperature. High temperatures above Tm led to at least partial dissociation of soluble filaments and thermal unfolding, resulting in the formation of soluble oligomers and binding to the persistently present soluble filaments. Such a complex formation between the oligomers and filaments has never been observed. Our results provide new insight into the heat-induced myosin gelation in high-salt solution. Copyright © 2014 Elsevier B.V. All rights reserved.

Rimmed vacuoles in Becker muscular dystrophy have similar features with inclusion myopathies.

PubMed

Momma, Kazunari; Noguchi, Satoru; Malicdan, May Christine V; Hayashi, Yukiko K; Minami, Narihiro; Kamakura, Keiko; Nonaka, Ikuya; Nishino, Ichizo

2012-01-01

Rimmed vacuoles in myofibers are thought to be due to the accumulation of autophagic vacuoles, and can be characteristic in certain myopathies with protein inclusions in myofibers. In this study, we performed a detailed clinical, molecular, and pathological characterization of Becker muscular dystrophy patients who have rimmed vacuoles in muscles. Among 65 Becker muscular dystrophy patients, we identified 12 patients who have rimmed vacuoles and 11 patients who have deletions in exons 45-48 in DMD gene. All patients having rimmed vacuoles showed milder clinical features compared to those without rimmed vacuoles. Interestingly, the rimmed vacuoles in Becker muscular dystrophy muscles seem to represent autophagic vacuoles and are also associated with polyubiquitinated protein aggregates. These findings support the notion that rimmed vacuoles can appear in Becker muscular dystrophy, and may be related to the chronic changes in muscle pathology induced by certain mutations in the DMD gene.

High-Spin Ferric Ions in Saccharomyces cerevisiae Vacuoles Are Reduced to the Ferrous State during Adenine-Precursor Detoxification

PubMed Central

2015-01-01

The majority of Fe in Fe-replete yeast cells is located in vacuoles. These acidic organelles store Fe for use under Fe-deficient conditions and they sequester it from other parts of the cell to avoid Fe-associated toxicity. Vacuolar Fe is predominantly in the form of one or more magnetically isolated nonheme high-spin (NHHS) FeIII complexes with polyphosphate-related ligands. Some FeIII oxyhydroxide nanoparticles may also be present in these organelles, perhaps in equilibrium with the NHHS FeIII. Little is known regarding the chemical properties of vacuolar Fe. When grown on adenine-deficient medium (A↓), ADE2Δ strains of yeast such as W303 produce a toxic intermediate in the adenine biosynthetic pathway. This intermediate is conjugated with glutathione and shuttled into the vacuole for detoxification. The iron content of A↓ W303 cells was determined by Mössbauer and EPR spectroscopies. As they transitioned from exponential growth to stationary state, A↓ cells (supplemented with 40 μM FeIII citrate) accumulated two major NHHS FeII species as the vacuolar NHHS FeIII species declined. This is evidence that vacuoles in A↓ cells are more reducing than those in adenine-sufficient cells. A↓ cells suffered less oxidative stress despite the abundance of NHHS FeII complexes; such species typically promote Fenton chemistry. Most Fe in cells grown for 5 days with extra yeast-nitrogen-base, amino acids and bases in minimal medium was HS FeIII with insignificant amounts of nanoparticles. The vacuoles of these cells might be more acidic than normal and can accommodate high concentrations of HS FeIII species. Glucose levels and rapamycin (affecting the TOR system) affected cellular Fe content. This study illustrates the sensitivity of cellular Fe to changes in metabolism, redox state and pH. Such effects broaden our understanding of how Fe and overall cellular metabolism are integrated. PMID:24919141

Immunofluorescent localization of ubiquitin and proteasomes in nucleolar vacuoles of soybean root meristematic cells

PubMed Central

Stępiński, D.

2012-01-01

In this study, using the immunofluorescent method, the immunopositive signals to ubiquitin and proteasomes in nucleoli of root meristematic cells of soybean seedlings have been observed. In fact, those signals were present exclusively in nucleolar vacuoles. No signals were observed in the nucleolar territory out of the nucleolar vacuoles or in the nucleoli without vacuoles. The ubiquitin-proteasome system (UPS) may act within the nucleoli of plants with high metabolic activities and may provide an additional level of regulation of intracellular proteolysis via compartment-specific activities of their components. It is suggested that the presence of the UPS solely in vacuolated nucleoli serves as a mechanism that enhances the speed of ribosome subunit production in very actively transcribing nucleoli. On the other hand, nucleolar vacuoles in a cell/nucleus could play additional roles associated with temporary sequestration or storage of some cellular factors, including components of the ubiquitin-proteasome system. PMID:22688294

Random myosin loss along thick-filaments increases myosin attachment time and the proportion of bound myosin heads to mitigate force decline in skeletal muscle

PubMed Central

Tanner, Bertrand C.W.; McNabb, Mark; Palmer, Bradley M.; Toth, Michael J.; Miller, Mark S.

2014-01-01

Diminished skeletal muscle performance with aging, disuse, and disease may be partially attributed to the loss of myofilament proteins. Several laboratories have found a disproportionate loss of myosin protein content relative to other myofilament proteins, but due to methodological limitations, the structural manifestation of this protein loss is unknown. To investigate how variations in myosin content affect ensemble cross-bridge behavior and force production we simulated muscle contraction in the half-sarcomere as myosin was removed either i) uniformly, from the Z-line end of thick-filaments, or ii) randomly, along the length of thick-filaments. Uniform myosin removal decreased force production, showing a slightly steeper force-to-myosin content relationship than the 1:1 relationship that would be expected from the loss of cross-bridges. Random myosin removal also decreased force production, but this decrease was less than observed with uniform myosin loss, largely due to increased myosin attachment time (ton) and fractional cross-bridge binding with random myosin loss. These findings support our prior observations that prolonged ton may augment force production in single fibers with randomly reduced myosin content from chronic heart failure patients. These simulation also illustrate that the pattern of myosin loss along thick-filaments influences ensemble cross-bridge behavior and maintenance of force throughout the sarcomere. PMID:24486373

Oriented thick and thin filaments in Amoeba proteus.

PubMed

Rinaldi, R A; Hrebenda, B

1975-07-01

Actin and myosin filaments as a foundation of contractile systems are well established from ameba to man (3). Wolpert et al. (19) isolated by differential centrifugation from Amoeba proteus a motile fraction composed of filaments which moved upon the addition of ATP. Actin filaments are found in amebas (1, 12, 13) which react with vertebrate heavy meromyosin (HMM), forming arrowhead complexes as vertebrate actin (3, 9), and are prominent within the ectoplasmic tube where some of them are attached to the plasmalemma (1, 12). Thick and thin filaments possessing the morphological characteristics of myosin and actin have been obtained from isolated ameba cytoplasm (18, 19). In addition, there are filaments exhibiting ATPase activity in amebas which react with actin (12, 16, 17). However, giant ameba (Chaos-proteus) shapes are difficult to preserve, and the excellent contributions referred to above are limited by visible distortions occurring in the amebas (rounding up, pseudopods disappearing, and cellular organelles swelling) upon fixation. Achievement of normal ameboid shape in recent glycerination work (15) led us to attempt other electron microscope fixation techniques, resulting in a surprising preservation of A. proteus with a unique orientation of thick and thin filaments in the ectoplasmic region.

Identification of Contractile Vacuole Proteins in Trypanosoma cruzi

PubMed Central

Park, Miyoung; Martins, Vicente P.; Atwood, James; Moles, Kristen; Collins, Dalis; Rohloff, Peter; Tarleton, Rick; Moreno, Silvia N. J.; Orlando, Ron; Docampo, Roberto

2011-01-01

Contractile vacuole complexes are critical components of cell volume regulation and have been shown to have other functional roles in several free-living protists. However, very little is known about the functions of the contractile vacuole complex of the parasite Trypanosoma cruzi, the etiologic agent of Chagas disease, other than a role in osmoregulation. Identification of the protein composition of these organelles is important for understanding their physiological roles. We applied a combined proteomic and bioinfomatic approach to identify proteins localized to the contractile vacuole. Proteomic analysis of a T. cruzi fraction enriched for contractile vacuoles and analyzed by one-dimensional gel electrophoresis and LC-MS/MS resulted in the addition of 109 newly detected proteins to the group of expressed proteins of epimastigotes. We also identified different peptides that map to at least 39 members of the dispersed gene family 1 (DGF-1) providing evidence that many members of this family are simultaneously expressed in epimastigotes. Of the proteins present in the fraction we selected several homologues with known localizations in contractile vacuoles of other organisms and others that we expected to be present in these vacuoles on the basis of their potential roles. We determined the localization of each by expression as GFP-fusion proteins or with specific antibodies. Six of these putative proteins (Rab11, Rab32, AP180, ATPase subunit B, VAMP1, and phosphate transporter) predominantly localized to the vacuole bladder. TcSNARE2.1, TcSNARE2.2, and calmodulin localized to the spongiome. Calmodulin was also cytosolic. Our results demonstrate the utility of combining subcellular fractionation, proteomic analysis, and bioinformatic approaches for localization of organellar proteins that are difficult to detect with whole cell methodologies. The CV localization of the proteins investigated revealed potential novel roles of these organelles in phosphate metabolism

UV/IR Filaments for High Resolution Novel Spectroscopic Interrogation of Plumes on Nuclear Materials

DTIC Science & Technology

2016-06-01

Understanding the physics and properties of high power filaments propagating in air, which include: (a) Femtosecond IR (mJ) filaments (b) Nanosecond UV (J...space and wavelength can resolve this controversy. The results have been published in Journal of Physics B, special issue on filamentation [19]. Only a...Raman scattering? 3. What is the impact of filamentation on the ratio of backward to forward Raman? 4. What is the physical origin/content of the

Mini-thin filaments regulated by troponin–tropomyosin

PubMed Central

Gong, Huiyu; Hatch, Victoria; Ali, Laith; Lehman, William; Craig, Roger; Tobacman, Larry S.

2005-01-01

Striated muscle thin filaments contain hundreds of actin monomers and scores of troponins and tropomyosins. To study the cooperative mechanism of thin filaments, “mini-thin filaments” were generated by isolating particles nearly matching the minimal structural repeat of thin filaments: a double helix of actin subunits with each strand approximately seven actins long and spanned by a troponin–tropomyosin complex. One end of the particles was capped by a gelsolin (segment 1–3)–TnT fusion protein (substituting for normal TnT), and the other end was capped by tropomodulin. EM showed that the particles were 46 ± 9 nm long, with a knob-like mass attributable to gelsolin at one end. Average actin, tropomyosin, and gelsolin–troponin composition indicated one troponin–tropomyosin attached to each strand of the two-stranded actin filament. The minifilaments thus nearly represent single regulatory units of thin filaments. The myosin S1 MgATPase rate stimulated by the minifilaments was Ca2+-sensitive, indicating that single regulatory length particles are sufficient for regulation. Ca2+ bound cooperatively to cardiac TnC in conventional thin filaments but noncooperatively to cardiac TnC in minifilaments in the absence of myosin. This suggests that thin filament Ca2+-binding cooperativity reflects indirect troponin–troponin interactions along the long axis of conventional filaments, which do not occur in minifilaments. Despite noncooperative Ca2+ binding to minifilaments in the absence of myosin, Ca2+ cooperatively activated the myosin S1-particle ATPase rate. Two-stranded single regulatory units therefore may be sufficient for myosin-mediated Ca2+-binding cooperativity. Functional mini-thin filaments are well suited for biochemical and structural analysis of thin-filament regulation. PMID:15644437

Pumping up the volume - vacuole biogenesis in Arabidopsis thaliana.

PubMed

Krüger, Falco; Schumacher, Karin

2017-07-08

Plant architecture follows the need to collect CO 2, solar energy, water and mineral nutrients via large surface areas. It is by the presence of a central vacuole that fills much of the cell volume that plants manage to grow at low metabolic cost. In addition vacuoles buffer the fluctuating supply of essential nutrients and help to detoxify the cytosol when plants are challenged by harmful molecules. Despite their large size and multiple important functions, our knowledge of vacuole biogenesis and the machinery underlying their amazing dynamics is still fragmentary. In this review, we try to reconcile past and present models for vacuole biogenesis with the current knowledge of multiple parallel vacuolar trafficking pathways and the molecular machineries driving membrane fusion and organelle shape. Copyright © 2017 Elsevier Ltd. All rights reserved.

Motility assays using myosin attached to surfaces through specific binding to monoclonal antibodies.

PubMed Central

Winkelmann, D. A.; Bourdieu, L.; Kinose, F.; Libchaber, A.

1995-01-01

We have analyzed the dependence of actin filament movement on the mode of myosin attachment to surfaces. Monoclonal antibodies that bind to three distinct sites were used to tether myosin to nitrocellulose-coated glass. One antibody reacts with an epitope on the regulatory light chain located at the head-rod junction. The other two react with sites in the rod domain, one in the S2 region near the S2-LMM hinge, and the other at the C terminus of the myosin rod. These monoclonal antibodies were used to provide increasing flexibility in the mode of attachment. Fast skeletal muscle myosin monomers were bound to the surfaces through the specific interaction with these monoclonal antibodies and the sliding movement of fluorescently labeled actin filaments analyzed by video microscopy. Each of these antibodies produced stable, myosin-coated surfaces that supported uniform movement of actin over the course of several hours. Attachment of myosin through the anti-S2 and anti-LMM monoclonal antibodies yielded a maximum velocity of 10 microns/s at 30 degrees C, whereas attachment through anti-LC2 produced a lower velocity of 4-5 microns/s. Each antibody showed a characteristic minimum myosin density below which sliding movement was no longer supported and an exponential dependence of actin filament velocity on myosin surface density below Vmax. Maximum sliding velocity was achieved over a range of myosin surface densities. Thus, the specific mode of attachment can influence the characteristic velocity of actin filament movement and the surface density needed to support movement. These data are being used to analyze the dynamics of sliding filament assays and evaluate estimates of the average number of motor molecules per unit length of actin required to support movement. PMID:7787107

Why Orange Guaymas Basin Beggiatoa spp. Are Orange: Single-Filament-Genome-Enabled Identification of an Abundant Octaheme Cytochrome with Hydroxylamine Oxidase, Hydrazine Oxidase, and Nitrite Reductase Activities

PubMed Central

Biddle, Jennifer F.; Siebert, Jason R.; Staunton, Eric; Hegg, Eric L.; Matthysse, Ann G.; Teske, Andreas

2013-01-01

Orange, white, and yellow vacuolated Beggiatoaceae filaments are visually dominant members of microbial mats found near sea floor hydrothermal vents and cold seeps, with orange filaments typically concentrated toward the mat centers. No marine vacuolate Beggiatoaceae are yet in pure culture, but evidence to date suggests they are nitrate-reducing, sulfide-oxidizing bacteria. The nearly complete genome sequence of a single orange Beggiatoa (“Candidatus Maribeggiatoa”) filament from a microbial mat sample collected in 2008 at a hydrothermal site in Guaymas Basin (Gulf of California, Mexico) was recently obtained. From this sequence, the gene encoding an abundant soluble orange-pigmented protein in Guaymas Basin mat samples (collected in 2009) was identified by microcapillary reverse-phase high-performance liquid chromatography (HPLC) nano-electrospray tandem mass spectrometry (μLC–MS-MS) of a pigmented band excised from a denaturing polyacrylamide gel. The predicted protein sequence is related to a large group of octaheme cytochromes whose few characterized representatives are hydroxylamine or hydrazine oxidases. The protein was partially purified and shown by in vitro assays to have hydroxylamine oxidase, hydrazine oxidase, and nitrite reductase activities. From what is known of Beggiatoaceae physiology, nitrite reduction is the most likely in vivo role of the octaheme protein, but future experiments are required to confirm this tentative conclusion. Thus, while present-day genomic and proteomic techniques have allowed precise identification of an abundant mat protein, and its potential activities could be assayed, proof of its physiological role remains elusive in the absence of a pure culture that can be genetically manipulated. PMID:23220958

Oriented thick and thin filaments in amoeba proteus

PubMed Central

Rinaldi, RA; Hrebenda, B

1975-01-01

Actin and myosin filaments as a foundation of contractile systems are well established from ameba to man (3). Wolpert et al. (19) isolated by differential centrifugation from Amoeba proteus a motile fraction composed of filaments which moved upon the addition of ATP. Actin filaments are found in amebas (1, 12, 13) which react with vertebrate heavy meromyosin (HMM), forming arrowhead complexes as vertebrate actin (3, 9), and are prominent within the ectoplasmic tube where some of them are attached to the plasmalemma (1, 12). Thick and thin filaments possessing the morphological characteristics of myosin and actin have been obtained from isolated ameba cytoplasm (18, 19). In addition, there are filaments exhibiting ATPase activity in amebas which react with actin (12, 16, 17). However, giant ameba (Chaos-proteus) shapes are difficult to preserve, and the excellent contributions referred to above are limited by visible distortions occurring in the amebas (rounding up, pseudopods disappearing, and cellular organelles swelling) upon fixation. Achievement of normal ameboid shape in recent glycerination work (15) led us to attempt other electron microscope fixation techniques, resulting in a surprising preservation of A. proteus with a unique orientation of thick and thin filaments in the ectoplasmic region. PMID:1141376

Topology of interaction between titin and myosin thick filaments.

PubMed

Kellermayer, Miklós; Sziklai, Dominik; Papp, Zsombor; Decker, Brennan; Lakatos, Eszter; Mártonfalvi, Zsolt

2018-05-05

Titin is a giant protein spanning between the Z- and M-lines of the sarcomere. In the A-band titin is associated with the myosin thick filament. It has been speculated that titin may serve as a blueprint for thick-filament formation due to the super-repeat structure of its A-band domains. Accordingly, titin might provide a template that determines the length and structural periodicity of the thick filament. Here we tested the titin ruler hypothesis by mixing titin and myosin at in situ stoichiometric ratios (300 myosins per 12 titins) in buffers of different ionic strength (KCl concentration range 100-300 mM). The topology of the filamentous complexes was investigated with atomic force microscopy. We found that the samples contained distinct, segregated populations of titin molecules and myosin thick filaments. We were unable to identify complexes in which myosin molecules were regularly associated to either mono- or oligomeric titin in either relaxed or stretched states of the titin filaments. Thus, the electrostatically driven self-association is stronger in both myosin and titin than their binding to each other, and it is unlikely that titin functions as a geometrical template for thick-filament formation. However, when allowed to equilibrate configurationally, long myosin thick filaments appeared with titin oligomers attached to their surface. The titin meshwork formed on the thick-filament surface may play a role in controlling thick-filament length by regulating the structural dynamics of myosin molecules and placing a mechanical limit on the filament length. Copyright © 2018 Elsevier Inc. All rights reserved.

Connections of intermediate filaments with the nuclear lamina and the cell periphery.

PubMed

Katsuma, Y; Swierenga, S H; Marceau, N; French, S W

1987-01-01

We investigated the relationship between intermediate filaments (IFs) and other detergent- and nuclease-resistant filamentous structures of cultured liver epithelial cells (T51B cell line) using whole mount unembedded preparations which were sequentially extracted with Triton X-100 and nucleases. Immunogold labelling and stereoscopic observation facilitated the examination of each filamentous structure and their three-dimensional relationships to each other. After solubilizing phospholipid, nucleic acid and soluble cellular protein, the resulting cytoskeleton preparation consisted of a network of cytokeratin and vimentin IFs linked by 3 nm filaments. The IFs were anchored to and determined the position of the nuclear lamina filaments (NLF) network and the centrioles. The NLF was composed of the nuclear lamina filaments measuring 3-6 nm in diameter which radiated from and anchored to the skeleton of the nuclear pores. The IFs located in the nuclear region appeared to be interwoven with the NLF. At the cell surface, the IFs seemed to be attached to the putative actin filament network. They formed a focally interrupted plexus-like structure at the cell periphery. Fragments of vimentin filaments were found among the filamentous network located at the cell surface, and some filaments terminated blindly there.

Sarcomere-length dependence of myosin filament structure in skeletal muscle fibres of the frog.

PubMed

Reconditi, Massimo; Brunello, Elisabetta; Fusi, Luca; Linari, Marco; Martinez, Manuel Fernandez; Lombardi, Vincenzo; Irving, Malcolm; Piazzesi, Gabriella

2014-03-01

X-ray diffraction patterns were recorded at beamline ID02 of the European Synchrotron Radiation Facility from small bundles of skeletal muscle fibres from Rana esculenta at sarcomere lengths between 2.1 and 3.5 μm at 4°C. The intensities of the X-ray reflections from resting fibres associated with the quasi-helical order of the myosin heads and myosin binding protein C (MyBP-C) decreased in the sarcomere length range 2.6-3.0 μm but were constant outside it, suggesting that an OFF conformation of the thick filament is maintained by an interaction between MyBP-C and the thin filaments. During active isometric contraction the intensity of the M3 reflection from the regular repeat of the myosin heads along the filaments decreased in proportion to the overlap between thick and thin filaments, with no change in its interference fine structure. Thus, myosin heads in the regions of the thick filaments that do not overlap with thin filaments are highly disordered during isometric contraction, in contrast to their quasi-helical order at rest. Heads in the overlap region that belong to two-headed myosin molecules that are fully detached from actin are also highly disordered, in contrast to the detached partners of actin-attached heads. These results provide strong support for the concept of a regulatory structural transition in the thick filament involving changes in both the organisation of the myosin heads on its surface and the axial periodicity of the myosin tails in its backbone, mediated by an interaction between MyBP-C and the thin filaments.

Sarcomere-length dependence of myosin filament structure in skeletal muscle fibres of the frog

PubMed Central

Reconditi, Massimo; Brunello, Elisabetta; Fusi, Luca; Linari, Marco; Martinez, Manuel Fernandez; Lombardi, Vincenzo; Irving, Malcolm; Piazzesi, Gabriella

2014-01-01

X-ray diffraction patterns were recorded at beamline ID02 of the European Synchrotron Radiation Facility from small bundles of skeletal muscle fibres from Rana esculenta at sarcomere lengths between 2.1 and 3.5 μm at 4°C. The intensities of the X-ray reflections from resting fibres associated with the quasi-helical order of the myosin heads and myosin binding protein C (MyBP-C) decreased in the sarcomere length range 2.6–3.0 μm but were constant outside it, suggesting that an OFF conformation of the thick filament is maintained by an interaction between MyBP-C and the thin filaments. During active isometric contraction the intensity of the M3 reflection from the regular repeat of the myosin heads along the filaments decreased in proportion to the overlap between thick and thin filaments, with no change in its interference fine structure. Thus, myosin heads in the regions of the thick filaments that do not overlap with thin filaments are highly disordered during isometric contraction, in contrast to their quasi-helical order at rest. Heads in the overlap region that belong to two-headed myosin molecules that are fully detached from actin are also highly disordered, in contrast to the detached partners of actin-attached heads. These results provide strong support for the concept of a regulatory structural transition in the thick filament involving changes in both the organisation of the myosin heads on its surface and the axial periodicity of the myosin tails in its backbone, mediated by an interaction between MyBP-C and the thin filaments. PMID:24344169

Thick filament mechano-sensing is a calcium-independent regulatory mechanism in skeletal muscle.

PubMed

Fusi, L; Brunello, E; Yan, Z; Irving, M

2016-10-31

Recent X-ray diffraction studies on actively contracting fibres from skeletal muscle showed that the number of myosin motors available to interact with actin-containing thin filaments is controlled by the stress in the myosin-containing thick filaments. Those results suggested that thick filament mechano-sensing might constitute a novel regulatory mechanism in striated muscles that acts independently of the well-known thin filament-mediated calcium signalling pathway. Here we test that hypothesis using probes attached to the myosin regulatory light chain in demembranated muscle fibres. We show that both the extent and kinetics of thick filament activation depend on thick filament stress but are independent of intracellular calcium concentration in the physiological range. These results establish direct control of myosin motors by thick filament mechano-sensing as a general regulatory mechanism in skeletal muscle that is independent of the canonical calcium signalling pathway.

Thick filament mechano-sensing is a calcium-independent regulatory mechanism in skeletal muscle

PubMed Central

Fusi, L.; Brunello, E.; Yan, Z.; Irving, M.

2016-01-01

Recent X-ray diffraction studies on actively contracting fibres from skeletal muscle showed that the number of myosin motors available to interact with actin-containing thin filaments is controlled by the stress in the myosin-containing thick filaments. Those results suggested that thick filament mechano-sensing might constitute a novel regulatory mechanism in striated muscles that acts independently of the well-known thin filament-mediated calcium signalling pathway. Here we test that hypothesis using probes attached to the myosin regulatory light chain in demembranated muscle fibres. We show that both the extent and kinetics of thick filament activation depend on thick filament stress but are independent of intracellular calcium concentration in the physiological range. These results establish direct control of myosin motors by thick filament mechano-sensing as a general regulatory mechanism in skeletal muscle that is independent of the canonical calcium signalling pathway. PMID:27796302

Additive Manufacturing of Multifunctional Components Using High Density Carbon Nanotube Yarn Filaments

NASA Technical Reports Server (NTRS)

Gardner, John M.; Sauti, Godfrey; Kim, Jae-Woo; Cano, Roberto J.; Wincheski, Russell A.; Stelter, Christopher J.; Grimsley, Brian W.; Working, Dennis C.; Siochi, Emilie J.

2016-01-01

Additive manufacturing allows for design freedom and part complexity not currently attainable using traditional manufacturing technologies. Fused Filament Fabrication (FFF), for example, can yield novel component geometries and functionalities because the method provides a high level of control over material placement and processing conditions. This is achievable by extrusion of a preprocessed filament feedstock material along a predetermined path. However if fabrication of a multifunctional part relies only on conventional filament materials, it will require a different material for each unique functionality printed into the part. Carbon nanotubes (CNTs) are an attractive material for many applications due to their high specific strength as well as good electrical and thermal conductivity. The presence of this set of properties in a single material presents an opportunity to use one material to achieve multifunctionality in an additively manufactured part. This paper describes a recently developed method for processing continuous CNT yarn filaments into three-dimensional articles, and summarizes the mechanical, electrical, and sensing performance of the components fabricated in this way.

A Rab-centric perspective of bacterial pathogen-occupied vacuoles.

PubMed

Sherwood, Racquel Kim; Roy, Craig R

2013-09-11

The ability to create and maintain a specialized organelle that supports bacterial replication is an important virulence property for many intracellular pathogens. Living in a membrane-bound vacuole presents inherent challenges, including the need to remodel a plasma membrane-derived organelle into a novel structure that will expand and provide essential nutrients to support replication, while also having the vacuole avoid membrane transport pathways that target bacteria for destruction in lysosomes. It is clear that pathogenic bacteria use different strategies to accomplish these tasks. The dynamics by which host Rab GTPases associate with pathogen-occupied vacuoles provide insight into the mechanisms used by different bacteria to manipulate host membrane transport. In this review we highlight some of the strategies bacteria use to maintain a pathogen-occupied vacuole by focusing on the Rab proteins involved in biogenesis and maintenance of these novel organelles. Copyright © 2013 Elsevier Inc. All rights reserved.

Rheology of Membrane-Attached Minimal Actin Cortices.

PubMed

Nöding, Helen; Schön, Markus; Reinermann, Corinna; Dörrer, Nils; Kürschner, Aileen; Geil, Burkhard; Mey, Ingo; Heussinger, Claus; Janshoff, Andreas; Steinem, Claudia

2018-04-26

The actin cortex is a thin cross-linked network attached to the plasma membrane, which is responsible for the cell's shape during migration, division, and growth. In a reductionist approach, we created a minimal actin cortex (MAC) attached to a lipid membrane to correlate the filamentous actin architecture with its viscoelastic properties. The system is composed of a supported 1-palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine bilayer doped with the receptor lipid phosphatidylinositol(4,5)-bisphosphate (PtdIns(4,5)P 2 ) to which a constitutively active mutant of ezrin, which is a direct membrane-cytoskeleton linker, is bound. The formation of the MAC on the supported lipid bilayer is analyzed as a function of increasing PtdIns(4,5)P 2 /ezrin pinning points, revealing an increase in the intersections between actin filaments, that is, the node density of the MAC. Bead tracking microrheology on the membrane-attached actin network provides information about its viscoelastic properties. The results show that ezrin serves as a dynamic cross-linker for the actin cortex attached to the lipid bilayer and that the stiffness of the network is influenced by the pinning point density, relating the plateau storage modulus G 0 to the node density of the MAC.

Filamentous actin organization in the unfertilized sea urchin egg cortex.

PubMed

Henson, J H; Begg, D A

1988-06-01

We have investigated the organization of filamentous actin in the cortex of unfertilized eggs of the sea urchins Strongylocentrotus purpuratus and Lytechinus variegatus. Rhodamine phalloidin and anti-actin immunofluorescent staining of isolated cortices reveal a punctate pattern of fluorescent sources. Comparison of this pattern with SEM images of microvillar morphology and distribution indicates that filamentous actin in the cortex is predominantly localized in the microvilli. Thin-section TEM and quick-freeze deep-etch ultrastructure of isolated cortices demonstrates that this microvillar-associated actin is in a novel organizational state composed of very short filaments arranged in a tight network and that these filament networks form mounds that extend beyond the plane of the plasma membrane. Actin filaments within the networks do not exhibit free ends and make end-on attachments with the membrane only within the region of the evaginating microvilli. Myosin S-1 dissociable crosslinks, 2-3 nm in diameter, are observed between network filaments and between network filaments and the membrane. A second population of long, individual actin filaments is observed in close lateral association with the plasma membrane and frequently complexes with the microvillar actin networks. The filamentous actin of the unfertilized egg cortex may participate in establishing the mechanical properties of the egg surface and may function in nucleating the assembly of cortical actin following fertilization.

Wortmannin-induced vacuole fusion enhances amyloplast dynamics in Arabidopsis zigzag1 hypocotyls

PubMed Central

Alvarez, Ashley Ann; Han, Sang Won; Toyota, Masatsugu; Brillada, Carla; Zheng, Jiameng; Gilroy, Simon

2016-01-01

Gravitropism in Arabidopsis shoots depends on the sedimentation of amyloplasts in the endodermis, and a complex interplay between the vacuole and F-actin. Gravity response is inhibited in zigzag-1 (zig-1), a mutant allele of VTI11, which encodes a SNARE protein involved in vacuole fusion. zig-1 seedlings have fragmented vacuoles that fuse after treatment with wortmannin, an inhibitor of phosphatidylinositol 3-kinase, and underscore a role of phosphoinositides in vacuole fusion. Using live-cell imaging with a vertical stage microscope, we determined that young endodermal cells below the apical hook that are smaller than 70 μm in length are the graviperceptive cells in dark-grown hypocotyls. This result was confirmed by local wortmannin application to the top of zig-1 hypocotyls, which enhanced shoot gravitropism in zig-1 mutants. Live-cell imaging of zig-1 hypocotyl endodermal cells indicated that amyloplasts are trapped between juxtaposed vacuoles and their movement is severely restricted. Wortmannin-induced fusion of vacuoles in zig-1 seedlings increased the formation of transvacuolar strands, enhanced amyloplast sedimentation and partially suppressed the agravitropic phenotype of zig-1 seedlings. Hypergravity conditions at 10 g were not sufficient to displace amyloplasts in zig-1, suggesting the existence of a physical tether between the vacuole and amyloplasts. Our results overall suggest that vacuole membrane remodeling may be involved in regulating the association of vacuoles and amyloplasts during graviperception. PMID:27816929

Flexibility of myosin attachment to surfaces influences F-actin motion.

PubMed Central

Winkelmann, D A; Bourdieu, L; Ott, A; Kinose, F; Libchaber, A

1995-01-01

We have analyzed the dependence of actin filament sliding movement on the mode of myosin attachment to surfaces. Monoclonal antibodies (mAbs) that bind to three distinct sites were used to tether myosin to nitrocellulose-coated glass. One antibody reacts with an epitope on the regulatory light chain (LC2) located at the head-rod junction. The other two react with sites in the rod domain, one in the S2 region near the S2-LMM hinge, and the other at the C terminus of the myosin rod. This method of attachment provides a means of controlling the flexibility and density of myosin on the surface. Fast skeletal muscle myosin monomers were bound to the surfaces through the specific interaction with these mAbs, and the sliding movement of fluorescently labeled actin filaments was analyzed by video microscopy. Each of these antibodies produced stable myosin-coated surfaces that supported uniform motion of actin over the course of several hours. Attachment of myosin through the anti-S2 and anti-LMM mAbs yielded significantly higher velocities (10 microns/s at 30 degrees C) than attachment through anti-LC2 (4-5 microns/s at 30 degrees C). For each antibody, we observed a characteristic value of the myosin density for the onset of F-actin motion and a second critical density for velocity saturation. The specific mode of attachment influences the velocity of actin filaments and the characteristic surface density needed to support movement. Images FIGURE 1 FIGURE 4 FIGURE 8 PMID:7544167

Formation, function, and exhaustion of notochordal cytoplasmic vacuoles within intervertebral disc: current understanding and speculation.

PubMed

Wang, Feng; Gao, Zeng-Xin; Cai, Feng; Sinkemani, Arjun; Xie, Zhi-Yang; Shi, Rui; Wei, Ji-Nan; Wu, Xiao-Tao

2017-08-22

Notochord nucleus pulposus cells are characteristic of containing abundant and giant cytoplasmic vacuoles. This review explores the embryonic formation, biological function, and postnatal exhaustion of notochord vacuoles, aiming to characterize the signal network transforming the vacuolated nucleus pulposus cells into the vacuole-less chondrocytic cells. Embryonically, the cytoplasmic vacuoles within vertebrate notochord originate from an evolutionarily conserved vacuolation process during neurulation, which may continue to provide mechanical and signal support in constructing a mammalian intervertebral disc. For full vacuolation, a vacuolating specification from dorsal organizer cells, synchronized convergent extension, well-structured notochord sheath, and sufficient post-Golgi trafficking in notochord cells are required. Postnatally, age-related and species-specific exhaustion of vacuolated nucleus pulposus cells could be potentiated by Fas- and Fas ligand-induced apoptosis, intolerance to mechanical stress and nutrient deficiency, vacuole-mediated proliferation check, and gradual de-vacuolation within the avascular and compression-loaded intervertebral disc. These results suggest that the notochord vacuoles are active and versatile organelles for both embryonic notochord and postnatal nucleus pulposus, and may provide novel information on intervertebral disc degeneration to guide cell-based regeneration.

High-resolution Observations of Downflows at One End of a Pre-eruption Filament

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Qin; Deng, Na; Jing, Ju

Studying the dynamics of filaments at the pre-eruption phase can shed light on the precursor of eruptive events. Such high-resolution studies (of the order of 0.″1) are highly desirable yet very rare. In this work, we present a detailed observation of a pre-eruption evolution of a filament obtained by the 1.6 m New Solar Telescope (NST) at the Big Bear Solar Observatory (BBSO). One end of the filament is anchored at the sunspot in the NOAA active region (AR) 11515, which is well observed by NST H α off-bands from four hours before to one hour after the filament eruption.more » A M1.6 flare is associated with the eruption. We observed persistent downflowing materials along the H α multi-threaded component of the loop toward the AR end during the pre-eruption phase. We traced the trajectories of plasma blobs along the H α threads and obtained a plane-of-sky velocity of 45 km s{sup −1} on average. Furthermore, we estimated the real velocities of the downflows and the altitude of the filament by matching the observed H α threads with magnetic field lines extrapolated from a nonlinear force-free field model. Observations of chromospheric brightenings at the footpoints of the falling plasma blobs are also presented. The lower limit of the kinetic energy per second of the downflows through the brightenings is found to be ∼10{sup 21} erg. Larger FOV observations from BBSO full-disk H α images show that the AR end of the filament started ascending four hours before the flare. We attribute the observed downflows at the AR end of the filament to the draining effect of the filament rising prior to its eruption. During the slow-rise phase, the downflows continuously drained away ∼10{sup 15}g mass from the filament over a few hours, which is believed to be essential for the instability, and could be an important precursor of eruptive events.« less

Shigella subverts the host recycling compartment to rupture its vacuole.

PubMed

Mellouk, Nora; Weiner, Allon; Aulner, Nathalie; Schmitt, Christine; Elbaum, Michael; Shorte, Spencer L; Danckaert, Anne; Enninga, Jost

2014-10-08

Shigella enters epithlial cells via internalization into a vacuole. Subsequent vacuolar membrane rupture allows bacterial escape into the cytosol for replication and cell-to-cell spread. Bacterial effectors such as IpgD, a PI(4,5)P2 phosphatase that generates PI(5)P and alters host actin, facilitate this internalization. Here, we identify host proteins involved in Shigella uptake and vacuolar membrane rupture by high-content siRNA screening and subsequently focus on Rab11, a constituent of the recycling compartment. Rab11-positive vesicles are recruited to the invasion site before vacuolar rupture, and Rab11 knockdown dramatically decreases vacuolar membrane rupture. Additionally, Rab11 recruitment is absent and vacuolar rupture is delayed in the ipgD mutant that does not dephosphorylate PI(4,5)P₂ into PI(5)P. Ultrastructural analyses of Rab11-positive vesicles further reveal that ipgD mutant-containing vacuoles become confined in actin structures that likely contribute to delayed vacular rupture. These findings provide insight into the underlying molecular mechanism of vacuole progression and rupture during Shigella invasion. Copyright © 2014 Elsevier Inc. All rights reserved.

Formation, function, and exhaustion of notochordal cytoplasmic vacuoles within intervertebral disc: current understanding and speculation

PubMed Central

Sinkemani, Arjun; Xie, Zhi-Yang; Shi, Rui; Wei, Ji-Nan; Wu, Xiao-Tao

2017-01-01

Notochord nucleus pulposus cells are characteristic of containing abundant and giant cytoplasmic vacuoles. This review explores the embryonic formation, biological function, and postnatal exhaustion of notochord vacuoles, aiming to characterize the signal network transforming the vacuolated nucleus pulposus cells into the vacuole-less chondrocytic cells. Embryonically, the cytoplasmic vacuoles within vertebrate notochord originate from an evolutionarily conserved vacuolation process during neurulation, which may continue to provide mechanical and signal support in constructing a mammalian intervertebral disc. For full vacuolation, a vacuolating specification from dorsal organizer cells, synchronized convergent extension, well-structured notochord sheath, and sufficient post-Golgi trafficking in notochord cells are required. Postnatally, age-related and species-specific exhaustion of vacuolated nucleus pulposus cells could be potentiated by Fas- and Fas ligand-induced apoptosis, intolerance to mechanical stress and nutrient deficiency, vacuole-mediated proliferation check, and gradual de-vacuolation within the avascular and compression-loaded intervertebral disc. These results suggest that the notochord vacuoles are active and versatile organelles for both embryonic notochord and postnatal nucleus pulposus, and may provide novel information on intervertebral disc degeneration to guide cell-based regeneration. PMID:28915712

Organelle Size Scaling of the Budding Yeast Vacuole by Relative Growth and Inheritance.

PubMed

Chan, Yee-Hung M; Reyes, Lorena; Sohail, Saba M; Tran, Nancy K; Marshall, Wallace F

2016-05-09

It has long been noted that larger animals have larger organs compared to smaller animals of the same species, a phenomenon termed scaling [1]. Julian Huxley proposed an appealingly simple model of "relative growth"-in which an organ and the whole body grow with their own intrinsic rates [2]-that was invoked to explain scaling in organs from fiddler crab claws to human brains. Because organ size is regulated by complex, unpredictable pathways [3], it remains unclear whether scaling requires feedback mechanisms to regulate organ growth in response to organ or body size. The molecular pathways governing organelle biogenesis are simpler than organogenesis, and therefore organelle size scaling in the cell provides a more tractable case for testing Huxley's model. We ask the question: is it possible for organelle size scaling to arise if organelle growth is independent of organelle or cell size? Using the yeast vacuole as a model, we tested whether mutants defective in vacuole inheritance, vac8Δ and vac17Δ, tune vacuole biogenesis in response to perturbations in vacuole size. In vac8Δ/vac17Δ, vacuole scaling increases with the replicative age of the cell. Furthermore, vac8Δ/vac17Δ cells continued generating vacuole at roughly constant rates even when they had significantly larger vacuoles compared to wild-type. With support from computational modeling, these results suggest there is no feedback between vacuole biogenesis rates and vacuole or cell size. Rather, size scaling is determined by the relative growth rates of the vacuole and the cell, thus representing a cellular version of Huxley's model. Copyright © 2016 Elsevier Ltd. All rights reserved.

Munc13-4 functions as a Ca2+ sensor for homotypic secretory granule fusion to generate endosomal exocytic vacuoles

PubMed Central

Woo, Sang Su; James, Declan J.; Martin, Thomas F. J.

2017-01-01

Munc13-4 is a Ca2+-dependent SNARE (soluble N-ethylmaleimide–sensitive factor attachment protein receptor)- and phospholipid-binding protein that localizes to and primes secretory granules (SGs) for Ca2+-evoked secretion in various secretory cells. Studies in mast cell–like RBL-2H3 cells provide direct evidence that Munc13–4 with its two Ca2+-binding C2 domains functions as a Ca2+ sensor for SG exocytosis. Unexpectedly, Ca2+ stimulation also generated large (>2.4 μm in diameter) Munc13-4+/Rab7+/Rab11+ endosomal vacuoles. Vacuole generation involved the homotypic fusion of Munc13-4+/Rab7+ SGs, followed by a merge with Rab11+ endosomes, and depended on Ca2+ binding to Munc13-4. Munc13-4 promoted the Ca2+-stimulated fusion of VAMP8-containing liposomes with liposomes containing exocytic or endosomal Q-SNAREs and directly interacted with late endosomal SNARE complexes. Thus Munc13-4 is a tethering/priming factor and Ca2+ sensor for both heterotypic SG-plasma membrane and homotypic SG-SG fusion. Total internal reflection fluorescence microscopy imaging revealed that vacuoles were exocytic and mediated secretion of β-hexosaminidase and cytokines accompanied by Munc13-4 diffusion onto the plasma membrane. The results provide new molecular insights into the mechanism of multigranular compound exocytosis commonly observed in various secretory cells. PMID:28100639

A high-content phenotypic screen reveals the disruptive potency of quinacrine and 3',4'-dichlorobenzamil on the digestive vacuole of Plasmodium falciparum.

PubMed

Lee, Yan Quan; Goh, Amanda S P; Ch'ng, Jun Hong; Nosten, François H; Preiser, Peter Rainer; Pervaiz, Shazib; Yadav, Sanjiv Kumar; Tan, Kevin S W

2014-01-01

Plasmodium falciparum is the etiological agent of malignant malaria and has been shown to exhibit features resembling programmed cell death. This is triggered upon treatment with low micromolar doses of chloroquine or other lysosomotrophic compounds and is associated with leakage of the digestive vacuole contents. In order to exploit this cell death pathway, we developed a high-content screening method to select compounds that can disrupt the parasite vacuole, as measured by the leakage of intravacuolar Ca(2+). This assay uses the ImageStream 100, an imaging-capable flow cytometer, to assess the distribution of the fluorescent calcium probe Fluo-4. We obtained two hits from a small library of 25 test compounds, quinacrine and 3',4'-dichlorobenzamil. The ability of these compounds to permeabilize the digestive vacuole in laboratory strains and clinical isolates was validated by confocal microscopy. The hits could induce programmed cell death features in both chloroquine-sensitive and -resistant laboratory strains. Quinacrine was effective at inhibiting field isolates in a 48-h reinvasion assay regardless of artemisinin clearance status. We therefore present as proof of concept a phenotypic screening method with the potential to provide mechanistic insights to the activity of antimalarial drugs.

Highs and Lows in English Attachment.

PubMed

Grillo, Nino; Costa, João; Fernandes, Bruno; Santi, Andrea

2015-11-01

Grillo and Costa (2014) claim that Relative-Clause attachment ambiguity resolution is largely dependent on whether or not a Pseudo-Relative interpretation is available. Data from Italian, and other languages allowing Pseudo-Relatives, support this hypothesis. Pseudo-Relative availability, however, covaries with the semantics of the main predicate (e.g., perceptual vs. stative). Experiment 1 assesses whether this predicate distinction alone can account for prior attachment results by testing it with a language that disallows Pseudo-Relatives (i.e. English). Low Attachment was found independent of Predicate-Type. Predicate-Type did however have a minor modulatory role. Experiment 2 shows that English, traditionally classified as a Low Attachment language, can demonstrate High Attachment with sentences globally ambiguous between a Small-Clause and a reduced Relative-Clause interpretation. These results support a grammatical account of previous effects and provide novel evidence for the parser's preference of a Small-Clause over a Restrictive interpretation, crosslinguistically. Copyright © 2015 Elsevier B.V. All rights reserved.

Synthesis and functionalization of coiled carbon filaments

NASA Astrophysics Data System (ADS)

Hikita, Muneaki

Coiled carbon filaments have one of the most attractive three-dimensional forms in carbon materials due to their helical morphologies. Because of their shape and carbon structure, they exhibit excellent mechanical and electrical properties such as superelasticity, low Young's modulus, relatively high electrical conductivity, and good electromagnetic (EM) wave absorption. Therefore, they are good candidates as fillers in composite materials for tactile sensor and electromagnetic interference shielding. In medical areas of interests, coiled carbon filaments can be used as micro and nano heaters or trigger for thermotherapy and biosensors using EM wave exposure because absorbed EM waves by coiled carbon filaments are converted into heat. Although various shapes of coiled carbon filaments have been discovered, optimum synthesis conditions and growth mechanisms of coiled carbon filaments are poorly understood. The study of growth kinetics is significant not only to analyze catalyst activity but also to establish the growth mechanisms of coiled carbon filaments. The establishment of growth mechanisms would be useful for determining optimum synthesis conditions and maximizing the quantity of carbon filaments synthesized for a given application. In the first study, tip grown single helical carbon filaments or carbon nanocoils (CNCs) were synthesized by a chemical vapor deposition method using tin-iron-oxide (Sn-Fe-O) xerogel film catalyst. The Sn-Fe-O catalyst was prepared by a low-cost sol-gel method using stannous acetate and ferric acetate as precursors. The growth kinetics of CNCs were monitored by a thermogravimetric analyzer, and the experimental result was correlated using a one-dimensional kinetic model, corresponding to one-dimensional tip growth. In the second study, bidirectionally grown double helical filaments or carbon microcoils (CMCs) were synthesized using a chemical vapor deposition method. CMCs obtained at two reaction temperatures were compared. CMCs

Extending Femtosecond Filamentation of High Power Laser Propagating in the Atmosphere

NASA Astrophysics Data System (ADS)

Eisenmann, Shmuel; Sivan, Yonatan; Fibich, Gadi; Zigler, Arie

2008-06-01

We show experimentally for ultrashort laser pulses propagating in air, that the filamentation distance of intense laser pulses in the atmosphere can be extended and controlled with a simple double-lens setup. Using this method we were able to achieve a 20-fold delay of the filamentation distance of non-chirped 120 fs pulses propagating in air, from 16 m to 330 m. At 330 m, the collapsing pulse is sufficiently powerful to create plasma filaments. We also show that the scatter of the filaments at 330 m can be significantly reduced by tilting the second lens. We derive a simple formula for the filamentation distance, and confirm its agreement with the experimental results. We also observe that delaying the onset of filamentation increases the filament length. To the best of our knowledge, this is the longest distance reported in the literature at which plasma filaments were created and controlled. Finally, we show that the peak power at the onset of collapse is significantly higher with the double-lens setup, compared with the standard negative chirping approach.

Galactic cold cores. VIII. Filament formation and evolution: Filament properties in context with evolutionary models

NASA Astrophysics Data System (ADS)

Rivera-Ingraham, A.; Ristorcelli, I.; Juvela, M.; Montillaud, J.; Men'shchikov, A.; Malinen, J.; Pelkonen, V.-M.; Marston, A.; Martin, P. G.; Pagani, L.; Paladini, R.; Paradis, D.; Ysard, N.; Ward-Thompson, D.; Bernard, J.-P.; Marshall, D. J.; Montier, L.; Tóth, L. V.

2017-05-01

Context. The onset of star formation is intimately linked with the presence of massive unstable filamentary structures. These filaments are therefore key for theoretical models that aim to reproduce the observed characteristics of the star formation process in the Galaxy. Aims: As part of the filament study carried out by the Herschel Galactic Cold Cores Key Programme, here we study and discuss the filament properties presented in GCC VII (Paper I) in context with theoretical models of filament formation and evolution. Methods: A conservatively selected sample of filaments located at a distance D< 500 pc was extracted from the GCC fields with the getfilaments algorithm. The physical structure of the filaments was quantified according to two main components: the central (Gaussian) region of the filament (core component), and the power-law-like region dominating the filament column density profile at larger radii (wing component). The properties and behaviour of these components relative to the total linear mass density of the filament and the column density of its environment were compared with the predictions from theoretical models describing the evolution of filaments under gravity-dominated conditions. Results: The feasibility of a transition from a subcritical to supercritical state by accretion at any given time is dependent on the combined effect of filament intrinsic properties and environmental conditions. Reasonably self-gravitating (high Mline,core) filaments in dense environments (AV≳ 3 mag) can become supercritical on timescales of t 1 Myr by accreting mass at constant or decreasing width. The trend of increasing Mline,tot (Mline,core and Mline,wing) and ridge AV with background for the filament population also indicates that the precursors of star-forming filaments evolve coevally with their environment. The simultaneous increase of environment and filament AV explains the observed association between dense environments and high Mline,core values

Process Properties of Electronic High Voltage Discharges Triggered by Ultra-short Pulsed Laser Filaments

NASA Astrophysics Data System (ADS)

Cvecek, Kristian; Gröschel, Benjamin; Schmidt, Michael

Remote processing of metallic workpieces by techniques based on electric arc discharge or laser irradiation for joining or cutting has a long tradition and is still being intensively investigated in present-day research. In applications that require high power processing, both approaches exhibit certain advantages and disadvantages that make them specific for a given task. While several hybrid approaches exist that try to combine the benefits of both techniques, none were as successful in providing a fixed electric discharge direction as discharges triggered by plasma filaments generated by ultra-short pulsed lasers. In this work we investigate spatial and temporal aspects of laser filament guided discharges and give an upper time delay between the filament creation and the electrical build-up of a dischargeable voltage for a successful filament triggered discharge.

Live morphological analysis of taxol-induced cytoplasmic vacuolization [corrected] in human lung adenocarcinoma cells.

PubMed

Wang, Xiao-Ping; Chen, Tong-Sheng; Sun, Lei; Cai, Ji-Ye; Wu, Ming-Qian; Mok, Martin

2008-12-01

Taxol (paclitaxel), one of the most active cancer chemotherapeutic agents, can cause programmed cell death (PCD) and cytoplasmic vacuolization. The objective of this study was to analyze the morphological characteristics induced by taxol. Human lung adenocarcinoma (ASTC-a-1) cells were exposed to various concentration of taxol. CCK-8 was used to assay the cell viability. Atomic force microscopy (AFM), plasmid transfection and confocal fluorescence microscopy were performed to image the cells morphological change induced by taxol. Fluorescence resonance energy transfer (FRET) was used to monitor the caspase-3 activation in living cells during taxol-induced cell death. Cells treated with taxol exhibited significant swelling and cytoplasmic vacuolization which may be due to endoplasmic reticulum (ER) vacuolization. Caspase-3 was not activated during taxol-induced cytoplasmic vacuolization and cell death. These findings suggest that taxol induces caspase-3-independent cytoplasmic vacuolization, cell swelling and cell death through ER vacuolization.

Novel Tonoplast Transporters Identified Using a Proteomic Approach with Vacuoles Isolated from Cauliflower Buds1[W][OA

PubMed Central

Schmidt, Ulrike G.; Endler, Anne; Schelbert, Silvia; Brunner, Arco; Schnell, Magali; Neuhaus, H. Ekkehard; Marty-Mazars, Daniéle; Marty, Francis; Baginsky, Sacha; Martinoia, Enrico

2007-01-01

Young meristematic plant cells contain a large number of small vacuoles, while the largest part of the vacuome in mature cells is composed by a large central vacuole, occupying 80% to 90% of the cell volume. Thus far, only a limited number of vacuolar membrane proteins have been identified and characterized. The proteomic approach is a powerful tool to identify new vacuolar membrane proteins. To analyze vacuoles from growing tissues we isolated vacuoles from cauliflower (Brassica oleracea) buds, which are constituted by a large amount of small cells but also contain cells in expansion as well as fully expanded cells. Here we show that using purified cauliflower vacuoles and different extraction procedures such as saline, NaOH, acetone, and chloroform/methanol and analyzing the data against the Arabidopsis (Arabidopsis thaliana) database 102 cauliflower integral proteins and 214 peripheral proteins could be identified. The vacuolar pyrophosphatase was the most prominent protein. From the 102 identified proteins 45 proteins were already described. Nine of these, corresponding to 46% of peptides detected, are known vacuolar proteins. We identified 57 proteins (55.9%) containing at least one membrane spanning domain with unknown subcellular localization. A comparison of the newly identified proteins with expression profiles from in silico data revealed that most of them are highly expressed in young, developing tissues. To verify whether the newly identified proteins were indeed localized in the vacuole we constructed and expressed green fluorescence protein fusion proteins for five putative vacuolar membrane proteins exhibiting three to 11 transmembrane domains. Four of them, a putative organic cation transporter, a nodulin N21 family protein, a membrane protein of unknown function, and a senescence related membrane protein were localized in the vacuolar membrane, while a white-brown ATP-binding cassette transporter homolog was shown to reside in the plasma membrane

Nano rare-earth oxides induced size-dependent vacuolization: an independent pathway from autophagy.

PubMed

Zhang, Ying; Yu, Chenguang; Huang, Guanyi; Wang, Changli; Wen, Longping

2010-09-07

Four rare earth oxides have been shown to induce autophagy. Interestingly, we often noticed plentiful vacuolization, which was not always involved in this autophagic process. In this study, we investigated three other rare-earth elements, including Yttrium (Y), Ytterbium (Yb), and Lanthanum (La). Autophagic effect could be induced by all of them but only Y(2)O(3) and Yb(2)O(3) could cause massive vacuolization. Y(2)O(3) and Yb(2)O(3) treated by sonication or centrifugation to reduce particle size were used to test vacuolization level in HeLa cell lines. The results showed that rare earth oxides-induced vacuolization is size-dependent and differs from autophagic pathway. To further clarify the characteristics of this autophagic process, we used MEF Atg-5 (autophagy associated gene 5) knockout cell line, and the result showed that the autophagic process induced by rare earth oxides is Atg-5-dependent and the observed vacuolization was independent from autophagy. Similar results could also be observed in our tests on 3-methyladenine(3-MA), a well-known autophagy inhibitor. In conclusion, for the first time, we clarified the relationship between massive vacuolization and autophagic process induced by rare earth oxides and pointed out the size effect of rare earth oxides on the formation of vacuoles, which give clues to further investigation on the mechanisms underlying their biological effects.

Nano rare-earth oxides induced size-dependent vacuolization: an independent pathway from autophagy

PubMed Central

Zhang, Ying; Yu, Chenguang; Huang, Guanyi; Wang, Changli; Wen, Longping

2010-01-01

Four rare earth oxides have been shown to induce autophagy. Interestingly, we often noticed plentiful vacuolization, which was not always involved in this autophagic process. In this study, we investigated three other rare-earth elements, including Yttrium (Y), Ytterbium (Yb), and Lanthanum (La). Autophagic effect could be induced by all of them but only Y2O3 and Yb2O3 could cause massive vacuolization. Y2O3 and Yb2O3 treated by sonication or centrifugation to reduce particle size were used to test vacuolization level in HeLa cell lines. The results showed that rare earth oxides-induced vacuolization is size-dependent and differs from autophagic pathway. To further clarify the characteristics of this autophagic process, we used MEF Atg-5 (autophagy associated gene 5) knockout cell line, and the result showed that the autophagic process induced by rare earth oxides is Atg-5-dependent and the observed vacuolization was independent from autophagy. Similar results could also be observed in our tests on 3-methyladenine(3-MA), a well-known autophagy inhibitor. In conclusion, for the first time, we clarified the relationship between massive vacuolization and autophagic process induced by rare earth oxides and pointed out the size effect of rare earth oxides on the formation of vacuoles, which give clues to further investigation on the mechanisms underlying their biological effects. PMID:20856835

Comparative analysis of Beggiatoa from hypersaline and marine environments.

PubMed

de Albuquerque, Julia Peixoto; Keim, Carolina Neumann; Lins, Ulysses

2010-07-01

The main criterion to classify a microorganism as belonging to the genus Beggiatoa is its morphology. All multicellular, colorless, gliding bacterial filaments containing sulfur globules described so far belong to this genus. At the ultrastructural level, they show also a very complex cell envelope structure. Here we describe uncultured vacuolated and non-vacuolated bacteria from two different environments showing all characteristics necessary to assign a bacterium to the genus Beggiatoa. We also intended to investigate whether narrow and vacuolate Beggiatoa do differ morphologically as much as they do phylogenetically. Both large, vacuolated trichomes and narrow filaments devoid of vacuoles were observed. We confirmed the identity of the narrow filaments by 16S rRNA phylogenetic analysis. The diameters of the trichomes ranged from 2.4 to 34 microm, and their lengths ranged from 10 microm to over 30 mm. Narrow trichomes moved by gliding at 3.0 microm/s; large filaments moved at 1.5 microm/s. Periplasmic sulfur inclusions were observed in both types of filaments, whereas phosphorus-rich bodies were found only in narrow trichomes. On the other hand, nitrate vacuoles were observed only in large trichomes. Ultra-thin section transmission electron microscopy showed differences between the cell ultrastructure of narrow (non-vacuolated) and large (vacuolated) Beggiatoa. We observed that cell envelopes from narrow Beggiatoa consist of five layers, whereas cell envelopes from large trichomes contain four layers. Copyright 2010 Elsevier Ltd. All rights reserved.

A STUDY OF HIGH MODULUS, HIGH STRENGTH FILAMENT MATERIALS BY DEPOSITION TECHNIQUES.

DTIC Science & Technology

had an average tensile strength of 240000 psi. The x-ray spectra of silicon carbide filaments made from SiCl4 , trichlorosilane, and...trichloromethylsilane were compared. The filaments produced from the two silane compounds showed strong peaks of SiC whereas the spectra for the SiCl4 produced

Transition of Femtosecond-Filament-Solid Interactions from Single to Multiple Filament Regime

DOE Office of Scientific and Technical Information (OSTI.GOV)

Skrodzki, P. J.; Burger, M.; Jovanovic, I.

High-peak-power fs-laser filaments offer unique characteristics attractive to remote sensing via techniques such as remote laser-induced breakdown spectroscopy (R-LIBS). The dynamics of several ablation mechanisms following the interaction between a filament and a solid determines the emission strength and reproducibility of target plasma, which is of relevance for R-LIBS applications. Here, we investigate the space- and time-resolved dynamics of ionic and atomic emission from copper as well as the surrounding atmosphere in order to understand limitations of fs-filament-ablation for standoff energy delivery. Furthermore, we probe the shock front produced from filament-target interaction using time-resolved shadowgraphy and infer laser-material coupling efficienciesmore » for both single and multiple filament regimes through analysis of shock expansion with the Sedov model for point detonation. The results provide insight into plasma structure for the range of peak powers up to 30 times the critical power for filamentation P cr. Despite the stochastic nucleation of multiple filaments at peak-powers greater than 16 P cr, emission of ionic and neutral species increases with pump beam intensity, and short-lived nitrogen emission originating from the ambient is consistently observed. Ultimately, results suggest favorable scaling of emission intensity from target species on the laser pump energy, furthering the prospects for use of filament-solid interactions for remote sensing.« less

Transition of Femtosecond-Filament-Solid Interactions from Single to Multiple Filament Regime

DOE PAGES

Skrodzki, P. J.; Burger, M.; Jovanovic, I.

2017-10-06

High-peak-power fs-laser filaments offer unique characteristics attractive to remote sensing via techniques such as remote laser-induced breakdown spectroscopy (R-LIBS). The dynamics of several ablation mechanisms following the interaction between a filament and a solid determines the emission strength and reproducibility of target plasma, which is of relevance for R-LIBS applications. Here, we investigate the space- and time-resolved dynamics of ionic and atomic emission from copper as well as the surrounding atmosphere in order to understand limitations of fs-filament-ablation for standoff energy delivery. Furthermore, we probe the shock front produced from filament-target interaction using time-resolved shadowgraphy and infer laser-material coupling efficienciesmore » for both single and multiple filament regimes through analysis of shock expansion with the Sedov model for point detonation. The results provide insight into plasma structure for the range of peak powers up to 30 times the critical power for filamentation P cr. Despite the stochastic nucleation of multiple filaments at peak-powers greater than 16 P cr, emission of ionic and neutral species increases with pump beam intensity, and short-lived nitrogen emission originating from the ambient is consistently observed. Ultimately, results suggest favorable scaling of emission intensity from target species on the laser pump energy, furthering the prospects for use of filament-solid interactions for remote sensing.« less

Macroautophagy and microautophagy in relation to vacuole formation in mesophyll cells of Dendrobium tepals.

PubMed

van Doorn, Wouter G; Kirasak, Kanjana; Ketsa, Saichol

2015-04-01

Prior to flower opening, mesophyll cells at the vascular bundles of Dendrobium tepals showed a large increase in vacuolar volume, partially at the expense of the cytoplasm. Electron micrographs indicated that this increase in vacuolar volume was mainly due to vacuole fusion. Macroautophagous structures typical of plant cells were observed. Only a small part of the decrease in cytoplasmic volume seemed due to macroautophagy. The vacuoles contained vesicles of various types, including multilamellar bodies. It was not clear if these vacuolar inclusions were due to macroautophagy or microautophagy. Only a single structure was observed of a protruding vacuole, indicating microautophagy. It is concluded that macroautophagy occurs in these cells but its role in vacuole formation seems small, while a possible role of microautophagy in vacuole formation might be hypothesized. Careful labeling of organelle membranes seems required to advance our insight in plant macro- and microautophagy and their roles in vacuole formation. Copyright © 2015 Elsevier GmbH. All rights reserved.

Calcium controls the formation of vacuoles from mitochondria to regulate microspore development in wheat.

PubMed

Li, Dong Xiao; Hu, Hai Yan; Li, Gan; Ru, Zhen Gang; Tian, Hui Qiao

2017-09-01

Potassium antimonite was used to investigate the localisation of calcium in developing wheat anthers to examine the relationship between Ca 2+ and pollen development. During anther development, calcium precipitate formation increased in anther wall cells prior to microspore mother cell meiosis and appeared in microspores, suggesting the presence of a calcium influx from anther wall cells into the locule. Initially, the precipitates in microspore cytoplasm primarily accumulated in the mitochondria and destroyed their inner membranes (cisterns) to become small vacuoles, which expanded and fused, ultimately becoming a large vacuole during microspore vacuolisation. After microspore division and large vacuole decomposition, many calcium precipitates again accumulated in the small vacuoles, indicating that calcium from the large vacuole moved back into the cytoplasm of bicellular pollen.

Toxoplasma gondii F-actin forms an extensive filamentous network required for material exchange and parasite maturation

PubMed Central

Periz, Javier; Whitelaw, Jamie; Harding, Clare; Gras, Simon; Del Rosario Minina, Mario Igor; Latorre-Barragan, Fernanda; Lemgruber, Leandro; Reimer, Madita Alice; Insall, Robert; Heaslip, Aoife; Meissner, Markus

2017-01-01

Apicomplexan actin is important during the parasite's life cycle. Its polymerization kinetics are unusual, permitting only short, unstable F-actin filaments. It has not been possible to study actin in vivo and so its physiological roles have remained obscure, leading to models distinct from conventional actin behaviour. Here a modified version of the commercially available actin-chromobody was tested as a novel tool for visualising F-actin dynamics in Toxoplasma gondii. Cb labels filamentous actin structures within the parasite cytosol and labels an extensive F-actin network that connects parasites within the parasitophorous vacuole and allows vesicles to be exchanged between parasites. In the absence of actin, parasites lack a residual body and inter-parasite connections and grow in an asynchronous and disorganized manner. Collectively, these data identify new roles for actin in the intracellular phase of the parasites lytic cycle and provide a robust new tool for imaging parasitic F-actin dynamics. DOI: http://dx.doi.org/10.7554/eLife.24119.001 PMID:28322189

Toxoplasma gondii F-actin forms an extensive filamentous network required for material exchange and parasite maturation.

PubMed

Periz, Javier; Whitelaw, Jamie; Harding, Clare; Gras, Simon; Del Rosario Minina, Mario Igor; Latorre-Barragan, Fernanda; Lemgruber, Leandro; Reimer, Madita Alice; Insall, Robert; Heaslip, Aoife; Meissner, Markus

2017-03-21

Apicomplexan actin is important during the parasite's life cycle. Its polymerization kinetics are unusual, permitting only short, unstable F-actin filaments. It has not been possible to study actin in vivo and so its physiological roles have remained obscure, leading to models distinct from conventional actin behaviour. Here a modified version of the commercially available actin-chromobody was tested as a novel tool for visualising F-actin dynamics in Toxoplasma gondii. Cb labels filamentous actin structures within the parasite cytosol and labels an extensive F-actin network that connects parasites within the parasitophorous vacuole and allows vesicles to be exchanged between parasites. In the absence of actin, parasites lack a residual body and inter-parasite connections and grow in an asynchronous and disorganized manner. Collectively, these data identify new roles for actin in the intracellular phase of the parasites lytic cycle and provide a robust new tool for imaging parasitic F-actin dynamics.

A filament of energetic particles near the high-latitude dawn magnetopause

NASA Technical Reports Server (NTRS)

Lui, A. T. Y.; Williams, D. J.; Mcentire, R. W.; Christon, S. P.; Jacquey, C.; Angelopoulos, V.; Yamamoto, T.; Kokubun, S.; Frank, L. A.; Ackerson, K. L.

1994-01-01

The Geotail satelite detected a filament of tailward-streaming energetic particles spatially separated from the boundary layer of energetic particles at the high-latitude dawn magnetopause at a downstream distance of approximately 80 R(sub E) on October 27, 1992. During this event, the composition and charge states of energetic ions at energies above approximately 10 keV show significant intermix of ions from solar wind and ionospheric sources. Detailed analysis leads to the deduction that the filament was moving southward towards the neutral sheet at an average speed of approximately 80 km/s, implying an average duskward electric field of approximately 1 mV/m. Its north-south dimension was approximately 1 R(sub E) and it was associated with an earthward directed field-aligned current of approximately 5 mA/m. The filament was separated from the energetic particle boundary layer straddling the magnetopause by approximately 0.8 R(sub E) and was inferred to be detached from the boundary layer at downstream distance beyond approximately 70 R(sub E) in the distant tail.

A proteomics dissection of Arabidopsis thaliana vacuoles isolated from cell culture

PubMed Central

Jaquinod, Michel; Villiers, Florent; Kieffer-Jaquinod, Sylvie; Hugouvieux, Véronique; Bruley, Christophe; Garin, Jérôme; Bourguignon, Jacques

2007-01-01

To better understand the mechanisms governing cellular traffic, storage of various metabolites and their ultimate degradation, Arabidopsis thaliana vacuoles proteomes were established. To this aim, a procedure was developed to prepare highly purified vacuoles from protoplasts isolated from Arabidopsis cell cultures using Ficoll density gradients. Based on the specific activity of the vacuolar marker α-mannosidase, the enrichment factor of the vacuoles was estimated at approximately 42 fold with an average yield of 2.1%. Absence of significant contamination by other cellular compartments was validated by western blot using antibodies raised against specific markers of chloroplasts, mitochondria, plasma membrane and endoplasmic reticulum. Based on these results, vacuole preparations showed the necessary degree of purity for proteomic study. Therefore, a proteomic approach was developed in order to identify the protein components present in both the membrane and soluble fractions of the Arabidopsis cell vacuoles. This approach includes: (i) a mild oxidation step leading to the transformation of cysteine residues into cysteic acid and methionine to methionine sulfoxide, (ii) an in-solution proteolytic digestion of very hydrophobic proteins, (iii) a pre-fractionation of proteins by short migration on SDS-PAGE followed by analysis by liquid chromatography coupled to tandem mass spectrometry. This procedure allowed the identification of more than 650 proteins, 2/3 of which copurify with the membrane hydrophobic fraction and 1/3 with the soluble fraction. Among the 416 proteins identified from the membrane fraction, 195 were considered integral membrane proteins based on the presence of one or more predicted transmembrane domains, and 110 transporters and related proteins were identified (91 putative transporters and 19 proteins related to the V-ATPase pump). With regard to function, about 20% of the proteins identified were previously known to be associated with vacuolar

Antimicrobial activity of filamentous fungi isolated from highly antibiotic-contaminated river sediment

PubMed Central

Svahn, K. Stefan; Göransson, Ulf; El-Seedi, Hesham; Bohlin, Lars; Larsson, D.G. Joakim; Olsen, Björn; Chryssanthou, Erja

2012-01-01

Background Filamentous fungi are well known for their production of substances with antimicrobial activities, several of which have formed the basis for the development of new clinically important antimicrobial agents. Recently, environments polluted with extraordinarily high levels of antibiotics have been documented, leading to strong selection pressure on local sentinel bacterial communities. In such microbial ecosystems, where multidrug-resistant bacteria are likely to thrive, it is possible that certain fungal antibiotics have become less efficient, thus encouraging alternative strategies for fungi to compete with bacteria. Methods In this study, sediment of a highly antibiotic-contaminated Indian river was sampled in order to investigate the presence of cultivable filamentous fungi and their ability to produce substances with antimicrobial activity. Results Sixty one strains of filamentous fungi, predominantly various Aspergillus spp. were identified. The majority of the Aspergillus strains displayed antimicrobial activity against methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamase-producing Escherichia coli, vancomycin-resistant Enterococcus faecalis and Candida albicans. Bioassay-guided isolation of the secondary metabolites of A. fumigatus led to the identification of gliotoxin. Conclusion This study demonstrated proof of principle of using bioassay-guided isolation for finding bioactive molecules. PMID:22957125

Multinodular and Vacuolating Neuronal Tumor: A Rare Seizure-associated Entity.

PubMed

Cathcart, Sahara J; Klug, Jeffrey R; Helvey, Jason T; L White, Matthew; Gard, Andrew P; McComb, Rodney D

2017-07-01

Multinodular and vacuolating neuronal tumor is a recently described seizure-associated entity with overlapping features of a malformative and neoplastic process. We report a case of multinodular and vacuolating neuronal tumor in a 29-year-old man with a history of recent headaches and complex partial seizures. Neuroimaging revealed a nonenhancing, T2 and T2 fluid-attenuated inversion recovery hyperintense multinodular lesion in the right temporal lobe. Lesional tissue demonstrated well-demarcated nodules of ganglioid cells with vacuolation of both the perikarya and the fibrillary neuropil-like background. The ganglioid cells showed weak cytoplasmic reactivity for synaptophysin and were nonreactive for neurofilament and chromogranin. CD34-positive stellate cells were present within the nodules. A 50-gene next-generation sequencing panel did not identify any somatic mutations in genomic DNA extracted from the tumor.

Multilamellar Structures and Filament Bundles Are Found on the Cell Surface during Bunyavirus Egress

PubMed Central

Sanz-Sánchez, Laura; Risco, Cristina

2013-01-01

Inside cells, viruses build specialized compartments for replication and morphogenesis. We observed that virus release associates with specific structures found on the surface of mammalian cells. Cultured adherent cells were infected with a bunyavirus and processed for oriented sectioning and transmission electron microscopy. Imaging of cell basal regions showed sophisticated multilamellar structures (MLS) and extracellular filament bundles with attached viruses. Correlative light and electron microscopy confirmed that both MLS and filaments proliferated during the maximum egress of new viruses. MLS dimensions and structure were reminiscent of those reported for the nanostructures on gecko fingertips, which are responsible for the extraordinary attachment capacity of these lizards. As infected cells with MLS were more resistant to detachment than control cells, we propose an adhesive function for these structures, which would compensate for the loss of adherence during release of new virus progeny. PMID:23799021

Trypanosoma cruzi Differentiates and Multiplies within Chimeric Parasitophorous Vacuoles in Macrophages Coinfected with Leishmania amazonensis

PubMed Central

Pessoa, Carina Carraro; Ferreira, Éden Ramalho; Bayer-Santos, Ethel; Rabinovitch, Michel; Mortara, Renato Arruda

2016-01-01

The trypanosomatids Leishmania amazonensis and Trypanosoma cruzi are excellent models for the study of the cell biology of intracellular protozoan infections. After their uptake by mammalian cells, the parasitic protozoan flagellates L. amazonensis and T. cruzi lodge within acidified parasitophorous vacuoles (PVs). However, whereas L. amazonensis develops in spacious, phagolysosome-like PVs that may enclose numerous parasites, T. cruzi is transiently hosted within smaller vacuoles from which it soon escapes to the host cell cytosol. To investigate if parasite-specific vacuoles are required for the survival and differentiation of T. cruzi, we constructed chimeric vacuoles by infection of L. amazonensis amastigote-infected macrophages with T. cruzi epimastigotes (EPIs) or metacyclic trypomastigotes (MTs). These chimeric vacuoles, easily observed by microscopy, allowed the entry and fate of T. cruzi in L. amazonensis PVs to be dynamically recorded by multidimensional imaging of coinfected cells. We found that although T. cruzi EPIs remained motile and conserved their morphology in chimeric vacuoles, T. cruzi MTs differentiated into amastigote-like forms capable of multiplying. These results demonstrate that the large adaptive vacuoles of L. amazonensis are permissive to T. cruzi survival and differentiation and that noninfective EPIs are spared from destruction within the chimeric PVs. We conclude that T. cruzi differentiation can take place in Leishmania-containing vacuoles, suggesting this occurs prior to their escape into the host cell cytosol. PMID:26975994

An electron microscope study of the contractile vacuole in Tokophrya infusionum.

PubMed

RUDZINSKA, M A

1958-03-25

Contractile vacuoles are organelles that collect fluid from the cytoplasm and expel it to the outside. After each discharge (systole), they appear again and expand (diastole). They are widely distributed among Protozoa, and have been found also in some fresh water algae, sponges, and recently in some blood cells of the frog, guinea pig, and man. In spite of the extensive work on the contractile vacuole, very little is known concerning its mode of operation. An electron microscope study of a suctorian Tokophrya infusionum provided an opportunity to study thin sections of contractile vacuoles, and in these some structures were found which could be part of a mechanism for the systolic and diastolic motions the organelle displays. In Tokophrya, as in Suctoria and Ciliata in general, the contractile vacuole has a permanent canal connecting it with the outside. The canal appears to have a very elaborate structure and is composed of three parts: (1) a pore; (2) a channel; and (3) a narrow tubule located in a papilla protruding into the cavity of the contractile vacuole. Whereas the pore and channel have fixed dimensions and are permanently widely open, the tubule has a changeable diameter. At diastole it is so narrow (about 25 to 30 mmicro in diameter) that it could be regarded as closed, while at systole it is widely open. It is assumed that the change in diameter is due to the contraction of numerous fine fibrils (about 180 A thick) which are radially disposed around the canal in form of a truncated cone, with its tip at the channel, and its base at the vacuolar membrane. It seems most probable that the broadening of the tubule results in discharge of the content of the contractile vacuole. In the vicinity of the very thin limiting vacuolar membrane, small vesicles and canaliculi of the endoplasmic reticulum, very small dense particles, and mitochondria may be found. In addition, rows of closely packed vesicles are present in this region, and in other parts of the

Direct visualization and in-depth physical study of metal filament formation in percolated high-κ dielectrics

NASA Astrophysics Data System (ADS)

Li, X.; Pey, K. L.; Bosman, M.; Liu, W. H.; Kauerauf, T.

2010-01-01

The migration of Ta atoms from a transistor gate electrode into the percolated high-κ (HK) gate dielectrics is directly shown using transmission electron microscopy analysis. A nanoscale metal filament that formed under high current injection is identified to be the physical defect responsible for the ultrafast transient breakdown (BD) of the metal-gate/high-κ (MG/HK) gate stacks. This highly conductive metal filament poses reliability concerns for MG/HK gate stacks as it significantly reduces the post-BD reliability margin of a transistor.

Aggregation of endosomal-vacuolar compartments in the Aovps24-deleted strain in the filamentous fungus Aspergillus oryzae

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tatsumi, Akinori; Shoji, Jun-ya; Kikuma, Takashi

2007-10-19

Previously, we found that deletion of Aovps24, an ortholog of Saccharomyces cerevisiae VPS24, that encodes an ESCRT (endosomal sorting complex required for transport)-III component required for late endosomal function results in fragmented and aggregated vacuoles. Although defective late endosomal function is likely responsible for this phenotype, critical lack of our knowledge on late endosomes in filamentous fungi prevented us from further characterization. In this study, we identified late endosomes of Aspergillus oryzae, by expressing a series of fusion proteins of fluorescent proteins with orthologs of late endosomal proteins. Using these fusion proteins as markers, we observed late endosomes in themore » wild type strain and the Aovps24 disruptant and demonstrated that late endosomes are aberrantly aggregated in the Aovps24 disruptant. Moreover, we revealed that the aggregated late endosomes have features of vacuoles as well. As deletion of another ESCRT-III component-encoding gene, Aovps2, resulted in similar phenotypes to that in the Aovps24 disruptant, phenotypes of the Aovps24 disruptant are probably due to defective late endosomal function.« less

Actin Hydrophobic Loop (262-274) and Filament Nucleation and Elongation

PubMed Central

Shvetsov, Alexander; Galkin, Vitold E.; Orlova, Albina; Phillips, Martin; Bergeron, Sarah E.; Rubenstein, Peter A.; Egelman, Edward H.; Reisler, Emil

2014-01-01

Summary The importance of actin hydrophobic loop 262-274 dynamics to actin polymerization and filament stability has been shown recently using a yeast actin mutant, L180C/L269C/C374A, in which the hydrophobic loop could be locked in a “parked” conformation by a disulfide bond between C180 and C269. Such a cross-linked G-actin does not form filaments, suggesting nucleation and/or elongation inhibition. To determine the role of loop dynamics in filament nucleation and/or elongation, we studied the polymerization of the cross-linked actin in the presence of cofilin - to assist with actin nucleation - and with phalloidin, to stabilize the elongating filament segments. We demonstrate here that together, but not alone, phalloidin and cofilin co-rescue the polymerization of cross-linked actin. The polymerization was also rescued by filament seeds added together with phalloidin but not with cofilin. Thus, loop immobilization via cross-linking inhibits both filament nucleation and elongation. Nevertheless, the conformational changes needed to catalyze ATP hydrolysis by actin occur in the cross-linked actin. When actin filaments are fully decorated by cofilin the helical twist of F-actin changes by ~ 5° per subunit. Electron microscopic analysis of filaments rescued by cofilin and phalloidin revealed a dense contact between opposite strands in F-actin, and a change of twist by ~ 1° per subunit, indicating either partial or disordered attachment of cofilin to F-actin and/or a competition between cofilin and phalloidin to alter F-actin symmetry. Our findings show an importance of the hydrophobic loop conformational dynamics to both actin nucleation and elongation and reveal that the inhibition of these two steps in the cross-linked actin can be relieved by appropriate factors. PMID:18037437

Morphology and kinematics of filaments in Serpens and Perseus molecular clouds: a high resolution study

NASA Astrophysics Data System (ADS)

Dhabal, Arnab; Mundy, Lee; Rizzo, Maxime; Storm, Shaye; Teuben, Peter; CLASSy Collaboration

2018-01-01

Filamentary structures are prevalent in molecular clouds over a wide range of scales, and are often associated with active star formation. The study of filament morphology and kinematics provide insights into the physical processes leading to core formation in clustered environments. As part of the CARMA Large Area Star Formation Survey (CLASSy) follow-up, we observed five Herschel filaments in the Serpens Main, Serpens South and NGC1333 molecular clouds using the J=1-0 transitions of dense gas tracers H13CO+, HNC and H13CN. Of these, H13CO+ and H13CN are optically thin and serve as a test of the kinematics previously seen by the CLASSy in N2H+. The observations have an angular resolution of 7'' and a spectral resolution of 0.16 km/s. Although the large scale structure compares well with the CARMA N2H+ (J=1-0) maps and Herschel dust continuum maps, we resolve finer structure within the filaments identified by Herschel. Most regions are found to have multiple structures and filaments partially overlapping in the line-of-sight. In two regions overlapping structures have velocity differences as high as 1.4 km/s. We identify 8 individual filaments with typical widths of 0.03-0.06 pc in these tracers, which is significantly less than widths observed in the Herschel dust column density maps. At least 50% of the filaments have distinct velocity gradients perpendicular to their major axis with average values in the range 4-10 km s-1 pc-1. These findings are in support of the theoretical models of filament formation by 2-D inflow in the shock layer created by colliding turbulent cells. We also find evidence of velocity gradients along the length of two filaments; the gradients suggest that these filaments are inflowing towards the cloud core.

Yeast Ivy1p Is a Putative I-BAR-domain Protein with pH-sensitive Filament Forming Ability in vitro.

PubMed

Itoh, Yuzuru; Kida, Kazuki; Hanawa-Suetsugu, Kyoko; Suetsugu, Shiro

2016-01-01

Bin-Amphiphysin-Rvs161/167 (BAR) domains mold lipid bilayer membranes into tubules, by forming a spiral polymer on the membrane. Most BAR domains are thought to be involved in forming membrane invaginations through their concave membrane binding surfaces, whereas some members have convex membrane binding surfaces, and thereby mold membranes into protrusions. The BAR domains with a convex surface form a subtype called the inverse BAR (I-BAR) domain or IRSp53-MIM-homology domain (IMD). Although the mammalian I-BAR domains have been studied, those from other organisms remain elusive. Here, we found putative I-BAR domains in Fungi and animal-like unicellular organisms. The fungal protein containing the putative I-BAR-domain is known as Ivy1p in yeast, and is reportedly localized in the vacuole. The phylogenetic analysis of the I-BAR domains revealed that the fungal I-BAR-domain containing proteins comprise a distinct group from those containing IRSp53 or MIM. Importantly, Ivy1p formed a polymer with a diameter of approximately 20 nm in vitro, without a lipid membrane. The filaments were formed at neutral pH, but disassembled when pH was reverted to basic. Moreover, Ivy1p and the I-BAR domain expressed in mammalian HeLa cells was localized at a vacuole-like structure as filaments as revealed by super-resolved microscopy. These data indicate the pH-sensitive polymer forming ability and the functional conservation of Ivy1p in eukaryotic cells.

Remote electrical arc suppression by laser filamentation.

PubMed

Schubert, Elise; Mongin, Denis; Kasparian, Jérôme; Wolf, Jean-Pierre

2015-11-02

We investigate the interaction of narrow plasma channels formed in the filamentation of ultrashort laser pulses, with a DC high voltage. The laser filaments prevent electrical arcs by triggering corona that neutralize the high-voltage electrodes. This phenomenon, that relies on the electric field modulation and free electron release around the filament, opens new prospects to lightning and over-voltage mitigation.

Aeromonas hydrophila exotoxin induces cytoplasmic vacuolation and cell death in VERO cells.

PubMed

Di Pietro, Angela; Picerno, Isa; Visalli, Giuseppa; Chirico, Cristina; Spataro, Pasquale; Cannavò, Giuseppe; Scoglio, Maria E

2005-07-01

Many organisms are able to cause cell vacuolation, but it is unclear if this can be considered a step of apoptosis or necrosis, or a distinct form of cell death. In this study VERO cells were used to evaluate the relationship between vacuolation and cell death pattern caused by exotoxins produced by environmental strains of A. hydrophila. Cell damage has been evaluated morphologically as well as biochemically. Cytotoxic and vacuolating titres were strictly correlated and the vacuolation has to be considered an early indicator of cytotoxicity that causes cell apoptosis or necrosis in relation to the dose. Signs of apoptosis (chromatin condensation and blebbing) were observed at low concentration and TGase activity, referable to apoptosis induction, confirms morphological observations. In fact, putrescine incorporation was related both to cytotoxin concentration and time of incubation. Moreover, the observed doubling cells with necrotic features permit us to suppose that cell sensitivity and death pattern could change during the different phases of cellular cycle.

The Water to Solute Permeability Ratio Governs the Osmotic Volume Dynamics in Beetroot Vacuoles.

PubMed

Vitali, Victoria; Sutka, Moira; Amodeo, Gabriela; Chara, Osvaldo; Ozu, Marcelo

2016-01-01

Plant cell vacuoles occupy up to 90% of the cell volume and, beyond their physiological function, are constantly subjected to water and solute exchange. The osmotic flow and vacuole volume dynamics relies on the vacuole membrane -the tonoplast- and its capacity to regulate its permeability to both water and solutes. The osmotic permeability coefficient ( P f ) is the parameter that better characterizes the water transport when submitted to an osmotic gradient. Usually, P f determinations are made in vitro from the initial rate of volume change, when a fast (almost instantaneous) osmolality change occurs. When aquaporins are present, it is accepted that initial volume changes are only due to water movements. However, in living cells osmotic changes are not necessarily abrupt but gradually imposed. Under these conditions, water flux might not be the only relevant driving force shaping the vacuole volume response. In this study, we quantitatively investigated volume dynamics of isolated Beta vulgaris root vacuoles under progressively applied osmotic gradients at different pH, a condition that modifies the tonoplast P f . We followed the vacuole volume changes while simultaneously determining the external osmolality time-courses and analyzing these data with mathematical modeling. Our findings indicate that vacuole volume changes, under progressively applied osmotic gradients, would not depend on the membrane elastic properties, nor on the non-osmotic volume of the vacuole, but on water and solute fluxes across the tonoplast. We found that the volume of the vacuole at the steady state is determined by the ratio of water to solute permeabilites ( P f / P s ), which in turn is ruled by pH. The dependence of the permeability ratio on pH can be interpreted in terms of the degree of aquaporin inhibition and the consequently solute transport modulation. This is relevant in many plant organs such as root, leaves, cotyledons, or stems that perform extensive rhythmic growth

The Water to Solute Permeability Ratio Governs the Osmotic Volume Dynamics in Beetroot Vacuoles

PubMed Central

Vitali, Victoria; Sutka, Moira; Amodeo, Gabriela; Chara, Osvaldo; Ozu, Marcelo

2016-01-01

Plant cell vacuoles occupy up to 90% of the cell volume and, beyond their physiological function, are constantly subjected to water and solute exchange. The osmotic flow and vacuole volume dynamics relies on the vacuole membrane -the tonoplast- and its capacity to regulate its permeability to both water and solutes. The osmotic permeability coefficient (Pf) is the parameter that better characterizes the water transport when submitted to an osmotic gradient. Usually, Pf determinations are made in vitro from the initial rate of volume change, when a fast (almost instantaneous) osmolality change occurs. When aquaporins are present, it is accepted that initial volume changes are only due to water movements. However, in living cells osmotic changes are not necessarily abrupt but gradually imposed. Under these conditions, water flux might not be the only relevant driving force shaping the vacuole volume response. In this study, we quantitatively investigated volume dynamics of isolated Beta vulgaris root vacuoles under progressively applied osmotic gradients at different pH, a condition that modifies the tonoplast Pf. We followed the vacuole volume changes while simultaneously determining the external osmolality time-courses and analyzing these data with mathematical modeling. Our findings indicate that vacuole volume changes, under progressively applied osmotic gradients, would not depend on the membrane elastic properties, nor on the non-osmotic volume of the vacuole, but on water and solute fluxes across the tonoplast. We found that the volume of the vacuole at the steady state is determined by the ratio of water to solute permeabilites (Pf/Ps), which in turn is ruled by pH. The dependence of the permeability ratio on pH can be interpreted in terms of the degree of aquaporin inhibition and the consequently solute transport modulation. This is relevant in many plant organs such as root, leaves, cotyledons, or stems that perform extensive rhythmic growth movements

Regulation of Contraction by the Thick Filaments in Skeletal Muscle.

PubMed

Irving, Malcolm

2017-12-19

Contraction of skeletal muscle cells is initiated by a well-known signaling pathway. An action potential in a motor nerve triggers an action potential in a muscle cell membrane, a transient increase of intracellular calcium concentration, binding of calcium to troponin in the actin-containing thin filaments, and a structural change in the thin filaments that allows myosin motors from the thick filaments to bind to actin and generate force. This calcium/thin filament mediated pathway provides the "START" signal for contraction, but it is argued that the functional response of the muscle cell, including the speed of its contraction and relaxation, adaptation to the external load, and the metabolic cost of contraction is largely determined by additional mechanisms. This review considers the role of the thick filaments in those mechanisms, and puts forward a paradigm for the control of contraction in skeletal muscle in which both the thick and thin filaments have a regulatory function. The OFF state of the thick filament is characterized by helical packing of most of the myosin head or motor domains on the thick filament surface in a conformation that makes them unavailable for actin binding or ATP hydrolysis, although a small fraction of the myosin heads are constitutively ON. The availability of the majority fraction of the myosin heads for contraction is controlled in part by the external load on the muscle, so that these heads only attach to actin and hydrolyze ATP when they are required. This phenomenon seems to be the major determinant of the well-known force-velocity relationship of muscle, and controls the metabolic cost of contraction. The regulatory state of the thick filament also seems to control the dynamics of both muscle activation and relaxation. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Method for fabricating ceramic filaments and high density tape casting method

NASA Technical Reports Server (NTRS)

Collins, Jr., Earl R. (Inventor)

1990-01-01

An apparatus and method is disclosed for fabricating mats of ceramic material comprising preparing a slurry of ceramic particles in a binder/solvent, charging the slurry into a vessel, forcing the slurry from the vessel into spinneret nozzles, discharging the slurry from the nozzles into the path of airjets to enhance the sinuous character of the slurry exudate and to dry it, collecting the filaments on a moving belt so that the filaments overlap each other thereby forming a mat, curing the binder therein, compressing and sintering the mat to form a sintered mat, and crushing the sintered mat to produce filament shaped fragments. A process is also disclosed for producing a tape of densely packed, bonded ceramic particles comprising forming a slurry of ceramic particles and a binder/solvent, applying the slurry to a rotating internal molding surface, applying a large centrifugal force to the slurry to compress it and force excess binder/solvent from the particles, evaporating solvent and curing the binder thereby forming layers of bonded ceramic particles and cured binder, and separating the binder layer from the layer of particles. Multilayers of ceramic particles are cast in an analogous manner on top of previously formed layers. When all of the desired layers have been cast the tape is fired to produce a sintered tape. For example, a three-layer tape is produced having outer layers of highly compressed filament shaped fragments of strontium doped lanthanum (LSM) particles and a center layer of yttria stabilized zicronia (YSZ) particles.

Electron emitting filaments for electron discharge devices

DOEpatents

Leung, Ka-Ngo; Pincosy, Philip A.; Ehlers, Kenneth W.

1988-01-01

Electrons are copiously emitted by a device comprising a loop-shaped filament made of lanthanum hexaboride. The filament is directly heated by an electrical current produced along the filament by a power supply connected to the terminal legs of the filament. To produce a filament, a diamond saw or the like is used to cut a slice from a bar made of lanthanum hexaboride. The diamond saw is then used to cut the slice into the shape of a loop which may be generally rectangular, U-shaped, hairpin-shaped, zigzag-shaped, or generally circular. The filaments provide high electron emission at a relatively low operating temperature, such as 1600.degree. C. To achieve uniform heating, the filament is formed with a cross section which is tapered between the opposite ends of the filament to compensate for non-uniform current distribution along the filament due to the emission of electrons from the filament.

Electron emitting filaments for electron discharge devices

DOEpatents

Leung, K.N.; Pincosy, P.A.; Ehlers, K.W.

1983-06-10

Electrons are copiously emitted by a device comprising a loop-shaped filament made of lanthanum hexaboride. The filament is directly heated by an electrical current produced along the filament by a power supply connected to the terminal legs of the filament. To produce a filament, a diamond saw or the like is used to cut a slice from a bar made of lanthanum hexaboride. The diamond saw is then used to cut the slice into the shape of a loop which may be generally rectangular, U-shaped, hairpin-shaped, zigzag-shaped, or generally circular. The filaments provide high electron emission at a relatively low operating temperature, such as 1600/sup 0/C. To achieve uniform heating, the filament is formed with a cross section which is tapered between the opposite ends of the filament to compensate for nonuniform current distribution along the filament due to the emission of electrons from the filament.

Simple fluorescence-based high throughput cell viability assay for filamentous fungi.

PubMed

Chadha, S; Kale, S P

2015-09-01

Filamentous fungi are important model organisms to understand the eukaryotic process and have been frequently exploited in research and industry. These fungi are also causative agents of serious diseases in plants and humans. Disease management strategies include in vitro susceptibility testing of the fungal pathogens to environmental conditions and antifungal agents. Conventional methods used for antifungal susceptibilities are cumbersome, time-consuming and are not suitable for a large-scale analysis. Here, we report a rapid, high throughput microplate-based fluorescence method for investigating the toxicity of antifungal and stress (osmotic, salt and oxidative) agents on Magnaporthe oryzae and compared it with agar dilution method. This bioassay is optimized for the resazurin reduction to fluorescent resorufin by the fungal hyphae. Resazurin bioassay showed inhibitory rates and IC50 values comparable to the agar dilution method and to previously reported IC50 or MICs for M. oryzae and other fungi. The present method can screen range of test agents from different chemical classes with different modes of action for antifungal activities in a simple, sensitive, time and cost effective manner. A simple fluorescence-based high throughput method is developed to test the effects of stress and antifungal agents on viability of filamentous fungus Magnaporthe oryzae. This resazurin fluorescence assay can detect inhibitory effects comparable to those obtained using the growth inhibition assay with added advantages of simplicity, time and cost effectiveness. This high throughput viability assay has a great potential in large-scale screening of the chemical libraries of antifungal agents, for evaluating the effects of environmental conditions and hyphal kinetic studies in mutant and natural populations of filamentous fungi. © 2015 The Society for Applied Microbiology.

Chirality and Magnetic Configurations of Solar Filaments

NASA Astrophysics Data System (ADS)

Ouyang, Y.; Zhou, Y. H.; Chen, P. F.; Fang, C.

2017-01-01

It has been revealed that the magnetic topology in the solar atmosphere displays hemispheric preference, I.e., helicity is mainly negative/positive in the northern/southern hemispheres, respectively. However, the strength of the hemispheric rule and its cyclic variation are controversial. In this paper, we apply a new method based on the filament drainage to 571 erupting filaments from 2010 May to 2015 December in order to determine the filament chirality and its hemispheric preference. It is found that 91.6% of our sample of erupting filaments follows the hemispheric rule of helicity sign. It is also found that the strength of the hemispheric preference of the quiescent filaments decreases slightly from ˜97% in the rising phase to ˜85% in the declining phase of solar cycle 24, whereas the strength of the intermediate filaments keeps a high value around 96 ± 4% at all times. Only the active-region filaments show significant variations. Their strength of the hemispheric rule rises from ˜63% to ˜95% in the rising phase, and keeps a high value of 82% ± 5% during the declining phase. Furthermore, during a half-year period around the solar maximum, their hemispheric preference totally vanishes. Additionally, we also diagnose the magnetic configurations of the filaments based on our indirect method and find that in our sample of erupting events, 89% are inverse-polarity filaments with a flux rope magnetic configuration, whereas 11% are normal-polarity filaments with a sheared arcade configuration.

Core filaments of the nuclear matrix

PubMed Central

1990-01-01

The nuclear matrix is concealed by a much larger mass of chromatin, which can be removed selectively by digesting nuclei with DNase I followed by elution of chromatin with 0.25 M ammonium sulfate. This mild procedure removes chromatin almost completely and preserves nuclear matrix morphology. The complete nuclear matrix consists of a nuclear lamina with an interior matrix composed of thick, polymorphic fibers and large masses that resemble remnant nucleoli. Further extraction of the nuclear matrices of HeLa or MCF-7 cells with 2 M sodium chloride uncovered a network of core filaments. A few dark masses remained enmeshed in the filament network and may be remnants of the nuclear matrix thick fibers and nucleoli. The highly branched core filaments had diameters of 9 and 13 nm measured relative to the intermediate filaments. They may serve as the core structure around which the matrix is constructed. The core filaments retained 70% of nuclear RNA. This RNA consisted both of ribosomal RNA precursors and of very high molecular weight hnRNA with a modal size of 20 kb. Treatment with RNase A removed the core filaments. When 2 M sodium chloride was used directly to remove chromatin after DNase I digestion without a preceding 0.25 M ammonium sulfate extraction, the core filaments were not revealed. Instead, the nuclear interior was filled with amorphous masses that may cover the filaments. This reflected a requirement for a stepwise increase in ionic strength because gradual addition of sodium chloride to a final concentration of 2 M without an 0.25 M ammonium sulfate extraction uncovered core filaments. PMID:2307700

Thick Filament Protein Network, Functions, and Disease Association.

PubMed

Wang, Li; Geist, Janelle; Grogan, Alyssa; Hu, Li-Yen R; Kontrogianni-Konstantopoulos, Aikaterini

2018-03-13

Sarcomeres consist of highly ordered arrays of thick myosin and thin actin filaments along with accessory proteins. Thick filaments occupy the center of sarcomeres where they partially overlap with thin filaments. The sliding of thick filaments past thin filaments is a highly regulated process that occurs in an ATP-dependent manner driving muscle contraction. In addition to myosin that makes up the backbone of the thick filament, four other proteins which are intimately bound to the thick filament, myosin binding protein-C, titin, myomesin, and obscurin play important structural and regulatory roles. Consistent with this, mutations in the respective genes have been associated with idiopathic and congenital forms of skeletal and cardiac myopathies. In this review, we aim to summarize our current knowledge on the molecular structure, subcellular localization, interacting partners, function, modulation via posttranslational modifications, and disease involvement of these five major proteins that comprise the thick filament of striated muscle cells. © 2018 American Physiological Society. Compr Physiol 8:631-709, 2018. Copyright © 2018 American Physiological Society. All rights reserved.

CHIRALITY AND MAGNETIC CONFIGURATIONS OF SOLAR FILAMENTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ouyang, Y.; Zhou, Y. H.; Chen, P. F.

It has been revealed that the magnetic topology in the solar atmosphere displays hemispheric preference, i.e., helicity is mainly negative/positive in the northern/southern hemispheres, respectively. However, the strength of the hemispheric rule and its cyclic variation are controversial. In this paper, we apply a new method based on the filament drainage to 571 erupting filaments from 2010 May to 2015 December in order to determine the filament chirality and its hemispheric preference. It is found that 91.6% of our sample of erupting filaments follows the hemispheric rule of helicity sign. It is also found that the strength of the hemisphericmore » preference of the quiescent filaments decreases slightly from ∼97% in the rising phase to ∼85% in the declining phase of solar cycle 24, whereas the strength of the intermediate filaments keeps a high value around 96 ± 4% at all times. Only the active-region filaments show significant variations. Their strength of the hemispheric rule rises from ∼63% to ∼95% in the rising phase, and keeps a high value of 82% ± 5% during the declining phase. Furthermore, during a half-year period around the solar maximum, their hemispheric preference totally vanishes. Additionally, we also diagnose the magnetic configurations of the filaments based on our indirect method and find that in our sample of erupting events, 89% are inverse-polarity filaments with a flux rope magnetic configuration, whereas 11% are normal-polarity filaments with a sheared arcade configuration.« less

Viruses of Entamoeba histolytica II. Morphogenesis of the Polyhedral Particle (ABRM2→HK-9)→HB-301 and the Filamentous Agent (ABRM)2→HK-9

PubMed Central

Mattern, Carl F. T.; Diamond, Louis S.; Daniel, Wendell A.

1972-01-01

The intracellular development of two morphologically different amoebal viruses has been studied by electron microscopy. One is a polyhedral agent which was observed as early as 24 hr after infection in the perinuclear cytoplasm. Subsequently, cell lysis occurred and particles were found in large number bound to membranes of disrupted amoebae. Other particles were found in phagocytic vacuoles suggesting a possible portal of entry into amoebae. The other virus is a filamentous particle which is first seen in small clusters in the nucleus after 24 hr of infection. The number of particles increases such that by 72 hr massive whorls of particles occupy a substantial part of the nucleus. After rupture of the nuclear membrane, clusters of filaments are widely dispersed throughout the cytoplasm. Still later, the cytoplasmic membrane disintegrates and clusters of filaments are found extracellularly, but free of cell membranes. The morphology of these agents is discussed in comparison with a variety of plant, animal, and bacterial viruses. Images PMID:4335523

Filaments, ridges and a mini-starburst - HOBYS' view of high mass star formation with Herschel

NASA Astrophysics Data System (ADS)

Hill, T.; Motte, F.; Didelon, P.

2012-03-01

With its unprecedented spatial resolution and high sensitivity, Herschel is revolutionising our understanding of high mass star formation and the interstellar medium (ISM). In particular, Herschel is unveiling the filamentary structure and molecular cloud constituents of the ISM where star formation takes place. The Herschel Imaging Survey of OB Young Stellar objects (HOBYS; Motte, Zavagno, Bontemps, see http://www.herschel.fr/cea/hobys/en/index.php) key program targets burgeoning young stellar objects with the aim of characterising them and the environments in which they form. HOBYS has already proven fruitful with many clear examples of high-mass star formation in nearby molecular cloud complexes (e.g. Motte et al., 2010). Through multi-wavelength Herschel observations I will introduce select regions of the HOBYS program, including Vela C, M16 and W48 to start. These data are rich with filamentary structures and a wealth of sources which span a large mass range including, low, intermediate and high-mass objects in the pre-collapse or protostellar phase of formation, many of which will proceed to form stars. The natal filaments themselves come in many shapes and sizes, they can form thick ridge-like structures, be dispersed in low column density regions or cluster in higher density regions. In Vela C, high-mass star formation proceeds preferentially in high column density supercritical filaments, called ridges, which may result from the constructive convergence of flows (Hill et al., 2011). I will present other examples of ridges identified in HOBYS regions. In addition, I will present the latest results on the Eagle Nebula (M16). This region was made iconic by Hubble, but only Herschel can trace the cold, dense early prestellar phases of star formation, and their natal interstellar filaments, in this infamous star-forming complex. The cavity ionised by the nearby OB cluster in M16 serves to heat the Pillars of Creation and the surrounding interstellar filaments

A soluble acid invertase is directed to the vacuole by a signal anchor mechanism.

PubMed

Rae, Anne L; Casu, Rosanne E; Perroux, Jai M; Jackson, Mark A; Grof, Christopher P L

2011-06-15

Enzyme activities in the vacuole have an important impact on the net concentration of sucrose. In sugarcane (Saccharum hybrid), immunolabelling demonstrated that a soluble acid invertase (β-fructofuranosidase; EC 3.2.1.26) is present in the vacuole of storage parenchyma cells during sucrose accumulation. Examination of sequences from sugarcane, barley and rice showed that the N-terminus of the invertase sequence contains a signal anchor and a tyrosine motif, characteristic of single-pass membrane proteins destined for lysosomal compartments. The N-terminal peptide from the barley invertase was shown to be capable of directing the green fluorescent protein to the vacuole in sugarcane cells. The results suggest that soluble acid invertase is sorted to the vacuole in a membrane-bound form. Copyright © 2010 Elsevier GmbH. All rights reserved.

Highly efficient broadband terahertz generation from ultrashort laser filamentation in liquids.

PubMed

Dey, Indranuj; Jana, Kamalesh; Fedorov, Vladimir Yu; Koulouklidis, Anastasios D; Mondal, Angana; Shaikh, Moniruzzaman; Sarkar, Deep; Lad, Amit D; Tzortzakis, Stelios; Couairon, Arnaud; Kumar, G Ravindra

2017-10-30

Generation and application of energetic, broadband terahertz pulses (bandwidth ~0.1-50 THz) is an active and contemporary area of research. The main thrust is toward the development of efficient sources with minimum complexities-a true table-top setup. In this work, we demonstrate the generation of terahertz radiation via ultrashort pulse induced filamentation in liquids-a counterintuitive observation due to their large absorption coefficient in the terahertz regime. The generated terahertz energy is more than an order of magnitude higher than that obtained from the two-color filamentation of air (the most standard table-top technique). Such high terahertz energies would generate electric fields of the order of MV cm -1 , which opens the doors for various nonlinear terahertz spectroscopic applications. The counterintuitive phenomenon has been explained via the solution of nonlinear pulse propagation equation in the liquid medium.

A First Approach to Filament Dynamics

ERIC Educational Resources Information Center

Silva, P. E. S.; de Abreu, F. Vistulo; Simoes, R.; Dias, R. G.

2010-01-01

Modelling elastic filament dynamics is a topic of high interest due to the wide range of applications. However, it has reached a high level of complexity in the literature, making it unaccessible to a beginner. In this paper we explain the main steps involved in the computational modelling of the dynamics of an elastic filament. We first derive…

High-throughput screening of filamentous fungi using nanoliter-range droplet-based microfluidics

NASA Astrophysics Data System (ADS)

Beneyton, Thomas; Wijaya, I. Putu Mahendra; Postros, Prexilia; Najah, Majdi; Leblond, Pascal; Couvent, Angélique; Mayot, Estelle; Griffiths, Andrew D.; Drevelle, Antoine

2016-06-01

Filamentous fungi are an extremely important source of industrial enzymes because of their capacity to secrete large quantities of proteins. Currently, functional screening of fungi is associated with low throughput and high costs, which severely limits the discovery of novel enzymatic activities and better production strains. Here, we describe a nanoliter-range droplet-based microfluidic system specially adapted for the high-throughput sceening (HTS) of large filamentous fungi libraries for secreted enzyme activities. The platform allowed (i) compartmentalization of single spores in ~10 nl droplets, (ii) germination and mycelium growth and (iii) high-throughput sorting of fungi based on enzymatic activity. A 104 clone UV-mutated library of Aspergillus niger was screened based on α-amylase activity in just 90 minutes. Active clones were enriched 196-fold after a single round of microfluidic HTS. The platform is a powerful tool for the development of new production strains with low cost, space and time footprint and should bring enormous benefit for improving the viability of biotechnological processes.

Interrelations between the Parasitophorous Vacuole of Toxoplasma gondii and Host Cell Organelles

NASA Astrophysics Data System (ADS)

Cardoso Magno, Rodrigo; Cobra Straker, Lorian; de Souza, Wanderley; Attias, Marcia

2005-04-01

Toxoplasma gondii, the causative agent of toxoplasmosis, is capable of actively penetrating and multiplying in any nucleated cell of warm-blooded animals. Its survival strategies include escape from fusion of the parasitophorous vacuole with host cell lysosomes and rearrangement of host cell organelles in relation to the parasitophorous vacuole. In this article we report the rearrangement of host cell organelles and elements of the cytoskeleton of LLCMK2 cells, a lineage derived from green monkey kidney epithelial cells, in response to infection by T. gondii tachyzoites. Transmission electron microscopy made on flat embedded monolayers cut horizontally to the apical side of the cells or field emission scanning electron microscopy of monolayers scraped with scotch tape before sputtering showed that association of mitochondria to the vacuole is much less frequent than previously described. On the other hand, all parasitophorous vacuoles were surrounded by elements of the endoplasmic reticulum. These data were complemented by observations by laser scanning microscopy using fluorescent probes from mitochondria and endoplasmic reticulum and reinforced by three-dimensional reconstruction from serial sections observed by transmission electron microscopy and labeling of mitochondria and endoplasmic reticulum by fluorescent probes.

The Metalloprotease Mpl Supports Listeria monocytogenes Dissemination through Resolution of Membrane Protrusions into Vacuoles

PubMed Central

Alvarez, Diego E.

2016-01-01

Listeria monocytogenes is an intracellular pathogen that disseminates within the intestinal epithelium through acquisition of actin-based motility and formation of plasma membrane protrusions that project into adjacent cells. The resolution of membrane protrusions into vacuoles from which the pathogen escapes results in bacterial spread from cell to cell. This dissemination process relies on the mlp-actA-plcB operon, which encodes ActA, a bacterial nucleation-promoting factor that mediates actin-based motility, and PlcB, a phospholipase that mediates vacuole escape. Here we investigated the role of the metalloprotease Mpl in the dissemination process. In agreement with previous findings showing that Mpl is required for PlcB activation, infection of epithelial cells with the ΔplcB or Δmpl strains resulted in the formation of small infection foci. As expected, the ΔplcB strain displayed a strong defect in vacuole escape. However, the Δmpl strain showed an unexpected defect in the resolution of protrusions into vacuoles, in addition to the expected but mild defect in vacuole escape. The Δmpl strain displayed increased levels of ActA on the bacterial surface in protrusions. We mapped an Mpl-dependent processing site in ActA between amino acid residues 207 to 238. Similar to the Δmpl strain, the ΔactA207–238 strain displayed increased levels of ActA on the bacterial surface in protrusions. Although the ΔactA207–238 strain displayed wild-type actin-based motility, it formed small infection foci and failed to resolve protrusions into vacuoles. We propose that, in addition to its role in PlcB processing and vacuole escape, the metalloprotease Mpl is required for ActA processing and protrusion resolution. PMID:27068088

The Metalloprotease Mpl Supports Listeria monocytogenes Dissemination through Resolution of Membrane Protrusions into Vacuoles.

PubMed

Alvarez, Diego E; Agaisse, Hervé

2016-06-01

Listeria monocytogenes is an intracellular pathogen that disseminates within the intestinal epithelium through acquisition of actin-based motility and formation of plasma membrane protrusions that project into adjacent cells. The resolution of membrane protrusions into vacuoles from which the pathogen escapes results in bacterial spread from cell to cell. This dissemination process relies on the mlp-actA-plcB operon, which encodes ActA, a bacterial nucleation-promoting factor that mediates actin-based motility, and PlcB, a phospholipase that mediates vacuole escape. Here we investigated the role of the metalloprotease Mpl in the dissemination process. In agreement with previous findings showing that Mpl is required for PlcB activation, infection of epithelial cells with the ΔplcB or Δmpl strains resulted in the formation of small infection foci. As expected, the ΔplcB strain displayed a strong defect in vacuole escape. However, the Δmpl strain showed an unexpected defect in the resolution of protrusions into vacuoles, in addition to the expected but mild defect in vacuole escape. The Δmpl strain displayed increased levels of ActA on the bacterial surface in protrusions. We mapped an Mpl-dependent processing site in ActA between amino acid residues 207 to 238. Similar to the Δmpl strain, the ΔactA207-238 strain displayed increased levels of ActA on the bacterial surface in protrusions. Although the ΔactA207-238 strain displayed wild-type actin-based motility, it formed small infection foci and failed to resolve protrusions into vacuoles. We propose that, in addition to its role in PlcB processing and vacuole escape, the metalloprotease Mpl is required for ActA processing and protrusion resolution. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Filament-strung stand-off elements for maintaining pane separation in vacuum insulating glazing units

DOEpatents

Bettger, Kenneth J; Stark, David H

2013-08-20

A vacuum insulating glazing unit (VIGU) comprises first and second panes of transparent material, first and second anchors, a plurality of filaments, a plurality of stand-off elements, and seals. The first and second panes of transparent material have edges and inner and outer faces, are disposed with their inner faces substantially opposing one another, and are separated by a gap having a predetermined height. The first and second anchors are disposed at opposite edges of one pane of the VIGU. Each filament is attached at one end to the first anchor and at the other end to the second anchor, and the filaments are collectively disposed between the panes substantially parallel to one another. The stand-off elements are affixed to each filament at predetermined positions along the filament, and have a height substantially equal to the predetermined height of the gap such that the each stand-off element touches the inner surfaces of both panes. The seals are disposed about the edges of the panes, enclosing the stand-off elements within a volume between the panes from which the atmosphere may be evacuated to form a partial vacuum.

Proglobulin processing enzyme in vacuoles isolated from developing pumpkin cotyledons

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hara-Nishimura, I.; Nishimura, M.

1987-10-01

The enzymic conversion of proglobulin to globulin catalyzed by the extracts of vacuoles isolated from developing pumpkin (Cucurbita sp. cv Kurokawa Amakuri Nankin) cotyledons was investigated. The endoplasmic reticulum fraction isolated from the developing cotyledons pulse-labeled with (/sup 35/S)methionine was shown to contain mainly the radiolabeled proglobulin, which was used as a substrate for assaying the proteolytic processing in vitro. The vacuolar extracts catalyzed the proteolytic processing of the proglobulin molecule to produce globulin containing two kinds of polypeptide chains, ..gamma.. and delta. The pH optimum for the vacuole-mediated conversion was at pH 5.0. The proteolytic processing of proglobulin bymore » the vacuolar extracts was inhibited in the presence of various thiol reagents, e.g. p-chloromercuribenzoate, N-ethylmaleimide, iodoacetic acid, Hg/sup 2 +/, and Cu/sup 2 +/, but not phenylmethylsulfonyl fluoride, EDTA, o-phenanthroline, leupeptin, antipain, pepstatin, chymostatin, or pumpkin trypsin inhibitor, and was activated in the presence of dithiothreitol and cysteine, indicating that the processing enzyme is a thiol protease. The suborganellar fractionation of the vacuoles showed that the processing activity was localized in the matrix fraction, but not in the membrane or crystalloid fractions. During the seed development, the enzyme was shown to increase, exhibiting the maximal activity at the late developmental stage. The matrix fraction of the protein bodies isolated from the dry castor bean (Ricinus communis) exhibited the processing activity toward the pumpkin proglobulin molecules in the same manner as that by the matrix fraction of pumpkin vacuoles.« less

Femtosecond Laser Filamentation for Atmospheric Sensing

PubMed Central

Xu, Huai Liang; Chin, See Leang

2011-01-01

Powerful femtosecond laser pulses propagating in transparent materials result in the formation of self-guided structures called filaments. Such filamentation in air can be controlled to occur at a distance as far as a few kilometers, making it ideally suited for remote sensing of pollutants in the atmosphere. On the one hand, the high intensity inside the filaments can induce the fragmentation of all matters in the path of filaments, resulting in the emission of characteristic fluorescence spectra (fingerprints) from the excited fragments, which can be used for the identification of various substances including chemical and biological species. On the other hand, along with the femtosecond laser filamentation, white-light supercontinuum emission in the infrared to UV range is generated, which can be used as an ideal light source for absorption Lidar. In this paper, we present an overview of recent progress concerning remote sensing of the atmosphere using femtosecond laser filamentation. PMID:22346566

In vitro assay using engineered yeast vacuoles for neuronal SNARE-mediated membrane fusion

PubMed Central

Ko, Young-Joon; Lee, Miriam; Kang, KyeongJin; Song, Woo Keun; Jun, Youngsoo

2014-01-01

Intracellular membrane fusion requires not only SNARE proteins but also other regulatory proteins such as the Rab and Sec1/Munc18 (SM) family proteins. Although neuronal SNARE proteins alone can drive the fusion between synthetic liposomes, it remains unclear whether they are also sufficient to induce the fusion of biological membranes. Here, through the use of engineered yeast vacuoles bearing neuronal SNARE proteins, we show that neuronal SNAREs can induce membrane fusion between yeast vacuoles and that this fusion does not require the function of the Rab protein Ypt7p or the SM family protein Vps33p, both of which are essential for normal yeast vacuole fusion. Although excess vacuolar SNARE proteins were also shown to mediate Rab-bypass fusion, this fusion required homotypic fusion and vacuole protein sorting complex, which bears Vps33p and was accompanied by extensive membrane lysis. We also show that this neuronal SNARE-driven vacuole fusion can be stimulated by the neuronal SM protein Munc18 and blocked by botulinum neurotoxin serotype E, a well-known inhibitor of synaptic vesicle fusion. Taken together, our results suggest that neuronal SNARE proteins are sufficient to induce biological membrane fusion, and that this new assay can be used as a simple and complementary method for investigating synaptic vesicle fusion mechanisms. PMID:24821814

Mutation-specific effects on thin filament length in thin filament myopathy.

PubMed

Winter, Josine M de; Joureau, Barbara; Lee, Eun-Jeong; Kiss, Balázs; Yuen, Michaela; Gupta, Vandana A; Pappas, Christopher T; Gregorio, Carol C; Stienen, Ger J M; Edvardson, Simon; Wallgren-Pettersson, Carina; Lehtokari, Vilma-Lotta; Pelin, Katarina; Malfatti, Edoardo; Romero, Norma B; Engelen, Baziel G van; Voermans, Nicol C; Donkervoort, Sandra; Bönnemann, C G; Clarke, Nigel F; Beggs, Alan H; Granzier, Henk; Ottenheijm, Coen A C

2016-06-01

Thin filament myopathies are among the most common nondystrophic congenital muscular disorders, and are caused by mutations in genes encoding proteins that are associated with the skeletal muscle thin filament. Mechanisms underlying muscle weakness are poorly understood, but might involve the length of the thin filament, an important determinant of force generation. We investigated the sarcomere length-dependence of force, a functional assay that provides insights into the contractile strength of muscle fibers as well as the length of the thin filaments, in muscle fibers from 51 patients with thin filament myopathy caused by mutations in NEB, ACTA1, TPM2, TPM3, TNNT1, KBTBD13, KLHL40, and KLHL41. Lower force generation was observed in muscle fibers from patients of all genotypes. In a subset of patients who harbor mutations in NEB and ACTA1, the lower force was associated with downward shifted force-sarcomere length relations, indicative of shorter thin filaments. Confocal microscopy confirmed shorter thin filaments in muscle fibers of these patients. A conditional Neb knockout mouse model, which recapitulates thin filament myopathy, revealed a compensatory mechanism; the lower force generation that was associated with shorter thin filaments was compensated for by increasing the number of sarcomeres in series. This allowed muscle fibers to operate at a shorter sarcomere length and maintain optimal thin-thick filament overlap. These findings might provide a novel direction for the development of therapeutic strategies for thin filament myopathy patients with shortened thin filament lengths. Ann Neurol 2016;79:959-969. © 2016 American Neurological Association.

Mutation-Specific Effects on Thin Filament Length in Thin Filament Myopathy

PubMed Central

de Winter, Josine M.; Joureau, Barbara; Lee, Eun-Jeong; Kiss, Balázs; Yuen, Michaela; Gupta, Vandana A.; Pappas, Christopher T.; Gregorio, Carol C.; Stienen, Ger J. M.; Edvardson, Simon; Wallgren-Pettersson, Carina; Lehtokari, Vilma-Lotta; Pelin, Katarina; Malfatti, Edoardo; Romero, Norma B.; van Engelen, Baziel G.; Voermans, Nicol C.; Donkervoort, Sandra; Bönnemann, C. G.; Clarke, Nigel F.; Beggs, Alan H.; Granzier, Henk; Ottenheijm, Coen A. C.

2016-01-01

Objective Thin filament myopathies are among the most common nondystrophic congenital muscular disorders, and are caused by mutations in genes encoding proteins that are associated with the skeletal muscle thin filament. Mechanisms underlying muscle weakness are poorly understood, but might involve the length of the thin filament, an important determinant of force generation. Methods We investigated the sarcomere length-dependence of force, a functional assay that provides insights into the contractile strength of muscle fibers as well as the length of the thin filaments, in muscle fibers from 51 patients with thin filament myopathy caused by mutations in NEB, ACTA1, TPM2, TPM3, TNNT1, KBTBD13, KLHL40, and KLHL41. Results Lower force generation was observed in muscle fibers from patients of all genotypes. In a subset of patients who harbor mutations in NEB and ACTA1, the lower force was associated with downward shifted force–sarcomere length relations, indicative of shorter thin filaments. Confocal microscopy confirmed shorter thin filaments in muscle fibers of these patients. A conditional Neb knockout mouse model, which recapitulates thin filament myopathy, revealed a compensatory mechanism; the lower force generation that was associated with shorter thin filaments was compensated for by increasing the number of sarcomeres in series. This allowed muscle fibers to operate at a shorter sarcomere length and maintain optimal thin–thick filament overlap. Interpretation These findings might provide a novel direction for the development of therapeutic strategies for thin filament myopathy patients with shortened thin filament lengths. PMID:27074222

Diagnosis of femtosecond plasma filament by channeling microwaves along the filament

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alshershby, Mostafa; Ren, Yu; Qin, Jiang

2013-05-20

We introduce a simple, fast, and non-intrusive experimental method to obtain the basic parameters of femtosecond laser-generated plasma filament. The method is based on the channeling of microwaves along both a plasma filament and a well-defined conducting wire. By comparing the detected microwaves that propagate along the plasma filament and a copper wire with known conductivity and spatial dimension, the basic parameters of the plasma filament can be easily obtained. As a result of the possibility of channeling microwave radiation along the plasma filament, we were then able to obtain the plasma density distribution along the filament length.

Bacterial filamentation accelerates colonization of adhesive spots embedded in biopassive surfaces

NASA Astrophysics Data System (ADS)

Möller, Jens; Emge, Philippe; Avalos Vizcarra, Ima; Kollmannsberger, Philip; Vogel, Viola

2013-12-01

Sessile bacteria adhere to engineered surfaces and host tissues and pose a substantial clinical and economical risk when growing into biofilms. Most engineered and biological interfaces are of chemically heterogeneous nature and provide adhesive islands for bacterial attachment and growth. To mimic either defects in a surface coating of biomedical implants or heterogeneities within mucosal layers (Peyer's patches), we embedded micrometre-sized adhesive islands in a poly(ethylene glycol) biopassive background. We show experimentally and computationally that filamentation of Escherichia coli can significantly accelerate the bacterial surface colonization under physiological flow conditions. Filamentation can thus provide an advantage to a bacterial population to bridge non-adhesive distances exceeding 5 μm. Bacterial filamentation, caused by blocking of bacterial division, is common among bacterial species and can be triggered by environmental conditions or antibiotic treatment. While great awareness exists that the build-up of antibiotic resistance serves as intrinsic survival strategy, we show here that antibiotic treatment can actually promote surface colonization by triggering filamentation, which in turn prevents daughter cells from being washed away. Our combined microfabrication and computational approaches provide quantitative insights into mechanisms that enable biofouling of biopassive surfaces with embedded adhesive spots, even for spot distances that are multiples of the bacterial length.

Pulsed, high-current, in-line reversal electron attachment detector

NASA Technical Reports Server (NTRS)

Bernius, Mark T.; Chutjian, Ara

1989-01-01

A new, pulsed, high-current, in-line reversal electron attachment ionizer/detector is described. The ionizer is capable of delivering a beam of electrons into an electrostatic mirror field to form a planar wall of electrons having zero kinetic energy. Electron attachment to a molecular target at the reversal point produces either parent or fragment negative ions through a zero-energy (s-wave) state. The atomic or molecular ion is pulsed out of the attachment region approximately 2 microsec after the electrons are pulsed off, and focused onto the entrance plane of a quadrupole mass analyzer. The sensitivity of the apparatus is preliminarily assessed, and its higher-energy behavior with regard to molecular attachment and ionization is described.

Chlamydia trachomatis Is Responsible for Lipid Vacuolation in the Amniotic Epithelium of Fetal Gastroschisis.

PubMed

Feldkamp, Marcia L; Ward, Diane M; Pysher, Theodore J; Chambers, Christina T

2017-07-17

Vacuolated amniotic epithelium with lipid droplets in gastroschisis placentas is an unusual finding. Mass spectrometry of lipid droplets identified triglycerides, ester-linked to an unusual pattern of fatty acids. We hypothesize that these findings result from a Chlamydia trachomatis infection during the periconceptional period. The rising incidence of chlamydia infections has paralleled the increasing prevalence of gastroschisis among women less than 25 years of age. Histologically, young women are at greatest risk for a chlamydia infection due to their immature columnar epithelium, the preferential site for attachment of Chlamydia trachomatis infectious particle (elementary body). Chlamydia trachomatis survive in an inclusion, relying on its host to acquire essential nutrients, amino acids, and nucleotides for survival and replication. If essential nutrients are not available, the bacteria cannot replicate and may be trafficked to the lysosome for degradation or remain quiescent, within the inclusion, subverting innate immunologic clearance. Chlamydiae synthesize several lipids (phosphatidylethanolamine, phosphatidylserine, and phosphoatidylglycerol); however, their lipid content reveal eukaryotic lipids (sphingomyelin, cholesterol, phosphatidylcholine, and phosphatidylinositol), evidence that chlamydiae "hijack" host lipids for expansion and replication. The abnormal amniotic epithelial findings are supported by experimental evidence of the trafficking of host lipids into the chlamydiae inclusion. If not lethal, what harm will elementary bodies inflict to the developing embryo? Do these women have a greater pro-inflammatory response to an environmental exposure, whether cigarette smoking, change in partner, or a pathogen? Testing the hypothesis that Chlamydia trachomatis is responsible for amniotic epithelium vacuoles will be a critical first step. Birth Defects Research 109:1003-1010, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Munc13-4 functions as a Ca2+ sensor for homotypic secretory granule fusion to generate endosomal exocytic vacuoles.

PubMed

Woo, Sang Su; James, Declan J; Martin, Thomas F J

2017-03-15

Munc13-4 is a Ca 2+ -dependent SNARE (soluble N -ethylmaleimide-sensitive factor attachment protein receptor)- and phospholipid-binding protein that localizes to and primes secretory granules (SGs) for Ca 2+ -evoked secretion in various secretory cells. Studies in mast cell-like RBL-2H3 cells provide direct evidence that Munc13-4 with its two Ca 2+ -binding C2 domains functions as a Ca 2+ sensor for SG exocytosis. Unexpectedly, Ca 2+ stimulation also generated large (>2.4 μm in diameter) Munc13-4 + /Rab7 + /Rab11 + endosomal vacuoles. Vacuole generation involved the homotypic fusion of Munc13-4 + /Rab7 + SGs, followed by a merge with Rab11 + endosomes, and depended on Ca 2+ binding to Munc13-4. Munc13-4 promoted the Ca 2+ -stimulated fusion of VAMP8-containing liposomes with liposomes containing exocytic or endosomal Q-SNAREs and directly interacted with late endosomal SNARE complexes. Thus Munc13-4 is a tethering/priming factor and Ca 2+ sensor for both heterotypic SG-plasma membrane and homotypic SG-SG fusion. Total internal reflection fluorescence microscopy imaging revealed that vacuoles were exocytic and mediated secretion of β-hexosaminidase and cytokines accompanied by Munc13-4 diffusion onto the plasma membrane. The results provide new molecular insights into the mechanism of multigranular compound exocytosis commonly observed in various secretory cells. © 2017 Woo et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Motor-mediated microtubule self-organization in dilute and semi-dilute filament solutions.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Swaminathan, S.; Ziebert, F.; Aranson, I. S.

We study molecular motor-induced microtubule self-organization in dilute and semi-dilute filament solutions. In the dilute case, we use a probabilistic model of microtubule interaction via molecular motors to investigate microtubule bundle dynamics. Microtubules are modeled as polar rods interacting through fully inelastic, binary collisions. Our model indicates that initially disordered systems of interacting rods exhibit an orientational instability resulting in spontaneous ordering. We study the existence and dynamic interaction of microtubule bundles analytically and numerically. Our results reveal a long term attraction and coalescing of bundles indicating a clear coarsening in the system; microtubule bundles concentrate into fewer orientations onmore » a slow logarithmic time scale. In semi-dilute filament solutions, multiple motors can bind a filament to several others and, for a critical motor density, induce a transition to an ordered phase with a nonzero mean orientation. Motors attach to a pair of filaments and walk along the pair bringing them into closer alignment. We develop a spatially homogenous, mean-field theory that explicitly accounts for a force-dependent detachment rate of motors, which in turn affects the mean and the fluctuations of the net force acting on a filament. We show that the transition to the oriented state can be both continuous and discontinuous when the force-dependent detachment of motors is important.« less

Intermediate filament mechanics in vitro and in the cell: from coiled coils to filaments, fibers and networks.

PubMed

Köster, Sarah; Weitz, David A; Goldman, Robert D; Aebi, Ueli; Herrmann, Harald

2015-02-01

Intermediate filament proteins form filaments, fibers and networks both in the cytoplasm and the nucleus of metazoan cells. Their general structural building plan accommodates highly varying amino acid sequences to yield extended dimeric α-helical coiled coils of highly conserved design. These 'rod' particles are the basic building blocks of intrinsically flexible, filamentous structures that are able to resist high mechanical stresses, that is, bending and stretching to a considerable degree, both in vitro and in the cell. Biophysical and computer modeling studies are beginning to unfold detailed structural and mechanical insights into these major supramolecular assemblies of cell architecture, not only in the 'test tube' but also in the cellular and tissue context. Copyright © 2015 Elsevier Ltd. All rights reserved.

MoVam7, a Conserved SNARE Involved in Vacuole Assembly, Is Required for Growth, Endocytosis, ROS Accumulation, and Pathogenesis of Magnaporthe oryzae

PubMed Central

Dou, Xianying; Wang, Qi; Qi, Zhongqiang; Song, Wenwen; Wang, Wei; Guo, Min; Zhang, Haifeng; Zhang, Zhengguang; Wang, Ping; Zheng, Xiaobo

2011-01-01

Soluble NSF attachment protein receptor (SNARE) proteins play a central role in membrane fusion and vesicle transport of eukaryotic organisms including fungi. We previously identified MoSce22 as a homolog of Saccharomyces cerevisiae SNARE protein Sec22 to be involved in growth, stress resistance, and pathogenicity of Magnaporthe oryzae. Here, we provide evidences that MoVam7, an ortholog of S. cerevisiae SNARE protein Vam7, exerts conserved functions in vacuolar morphogenesis and functions in pathogenicity of M. oryzae. Staining with neutral red and FM4-64 revealed the presence of abnormal fragmented vacuoles and an absence of the Spitzenkörper body in the ΔMovam7 mutant. The ΔMovam7 mutant also exhibited reduced vegetative growth, poor conidiation, and failure to produce the infection structure appressorium. Additionally, treatments with cell wall perturbing agents indicated weakened cell walls and altered distributions of the cell wall component chitin. Furthermore, the ΔMovam7 mutant showed a reduced accumulation of reactive oxygen species (ROS) in the hyphal apex and failed to cause diseases on the rice plant. In summary, our studies indicate that MoVam7, like MoSec22, is a component of the SNARE complex whose functions in vacuole assembly also underlies the growth, conidiation, appressorium formation, and pathogenicity of M. oryzae. Further studies of MoVam7, MoSec22, and additional members of the SNARE complex are likely to reveal critical mechanisms in vacuole formation and membrane trafficking that is linked to fungal pathogenicity. PMID:21283626

Arabidopsis ribosomal proteins control vacuole trafficking and developmental programs through the regulation of lipid metabolism.

PubMed

Li, Ruixi; Sun, Ruobai; Hicks, Glenn R; Raikhel, Natasha V

2015-01-06

The vacuole is the most prominent compartment in plant cells and is important for ion and protein storage. In our effort to search for key regulators in the plant vacuole sorting pathway, ribosomal large subunit 4 (rpl4d) was identified as a translational mutant defective in both vacuole trafficking and normal development. Polysome profiling of the rpl4d mutant showed reduction in polysome-bound mRNA compared with wild-type, but no significant change in the general mRNA distribution pattern. Ribsomal profiling data indicated that genes in the lipid metabolism pathways were translationally down-regulated in the rpl4d mutant. Live imaging studies by Nile red staining suggested that both polar and nonpolar lipid accumulation was reduced in meristem tissues of rpl4d mutants. Pharmacological evidence showed that sterol and sphingolipid biosynthetic inhibitors can phenocopy the defects of the rpl4d mutant, including an altered vacuole trafficking pattern. Genetic evidence from lipid biosynthetic mutants indicates that alteration in the metabolism of either sterol or sphingolipid biosynthesis resulted in vacuole trafficking defects, similar to the rpl4d mutant. Tissue-specific complementation with key enzymes from lipid biosynthesis pathways can partially rescue both vacuole trafficking and auxin-related developmental defects in the rpl4d mutant. These results indicate that lipid metabolism modulates auxin-mediated tissue differentiation and endomembrane trafficking pathways downstream of ribosomal protein function.

Filamentation and spatiotemporal distribution of extracellular polymeric substances: role on X.fastidiosa single cell adhesion and biofilm formation (Conference Presentation)

NASA Astrophysics Data System (ADS)

Janissen, Richard; Murillo, Duber M.; Niza, Barbara; Sahoo, Prasana K.; Monteiro, Moniellen P.; César, Carlos L.; Carvalho, Hernandes F.; de Souza, Alessandra A.; Cotta, Monica A.

2016-04-01

Biofilms can be defined as a community of microorganisms attached to a surface, living embedded in a self- produced matrix of hydrated extracellular polymeric substances (EPS) which comprises most of the biofilm mass. We have recently used an extensive pool of microscopy techniques (confocal fluorescence, electron and scanning probe microscopies) at the micro and nanoscales in order to create a detailed temporal observation of Xylella fastidiosa biofilm formation, using both wild type strain and Green Fluorescent Protein (GFP)-modified cells of this citrus phytopathogen. We have identified three different EPS compositions, as well as their spatial and temporal distribution from single cell to mature biofilm formation stages. In the initial adhesion stage, soluble-EPS (S-EPS) accumulates at cell polar regions and forms a surface layer which facilitates irreversible cell attachment and cell cluster formation. These small clusters are subsequently connected by filamentous cells; further S-EPS surface coverage facilitates cell attachment and form filaments, leading to a floating framework of mature biofilms. The important role of EPS in X.fastidiosa biology was further investigated by imunolabelling experiments to detect the distribution of XadA1 adhesin, which is expressed in early stages of biofilm formation and released in outer membrane vesicles. This protein is located mainly in S-EPS covered areas, as well as on the filaments, indicating a molecular pathway to the enhanced cell attachment previously observed. These results suggest that S-EPS may thus represent an important target for disease control, slow plant colonization by the bacteria, keeping the plant more productive in the field.

Post-filamentation high-intensive light channels formation upon ultrashort laser pulses self-focusing in air

NASA Astrophysics Data System (ADS)

Geints, Yu. E.; Ionin, A. A.; Mokrousova, D. V.; Seleznev, L. V.; Sinitsyn, D. V.; Sunchugasheva, E. S.; Zemlyanov, A. A.

2017-01-01

Experimental and theoretical study of the post-filamentation stage of focused high-power Ti:Sa laser pulses in air is presented. Angular divergence of the laser beam, as well as angular and spatial characteristics of specific spatially localized light structures, the post-filament channels (PFCs), under different initial focusing conditions and laser beam energy are investigated. We show that PFC angular divergence is always less than that of the whole laser beam and tends to decrease with laser pulse energy increase and beam focal length elongation.

Aerogel-supported filament

DOEpatents

Wuest, Craig R.; Tillotson, Thomas M.; Johnson, III, Coleman V.

1995-01-01

The present invention is a thin filament embedded in a low density aerogel for use in radiation detection instruments and incandescent lamps. The aerogel provides a supportive matrix that is thermally and electrically nonconductive, mechanically strong, highly porous, gas-permeable, and transparent to ionizing radiation over short distances. A low density, open-cell aerogel is cast around a fine filament or wire, which allows the wire to be positioned with little or no tension and keeps the wire in place in the event of breakage. The aerogel support reduces the stresses on the wire caused by vibrational, gravitational, electrical, and mechanical forces.

Vacuolar respiration of nitrate coupled to energy conservation in filamentous Beggiatoaceae.

PubMed

Beutler, Martin; Milucka, Jana; Hinck, Susanne; Schreiber, Frank; Brock, Jörg; Mussmann, Marc; Schulz-Vogt, Heide N; de Beer, Dirk

2012-11-01

We show that the nitrate storing vacuole of the sulfide-oxidizing bacterium Candidatus Allobeggiatoa halophila has an electron transport chain (ETC), which generates a proton motive force (PMF) used for cellular energy conservation. Immunostaining by antibodies showed that cytochrome c oxidase, an ETC protein and a vacuolar ATPase are present in the vacuolar membrane and cytochrome c in the vacuolar lumen. The effect of different inhibitors on the vacuolar pH was studied by pH imaging. Inhibition of vacuolar ATPases and pyrophosphatases resulted in a pH decrease in the vacuole, showing that the proton gradient over the vacuolar membrane is used for ATP and pyrophosphate generation. Blockage of the ETC decreased the vacuolar PMF, indicating that the proton gradient is build up by an ETC. Furthermore, addition of nitrate resulted in an increase of the vacuolar PMF. Inhibition of nitrate reduction, led to a decreased PMF. Nitric oxide was detected in vacuoles of cells exposed to nitrate showing that nitrite, the product of nitrate reduction, is reduced inside the vacuole. These findings show consistently that nitrate respiration contributes to the high proton concentration within the vacuole and the PMF over the vacuolar membrane is actively used for energy conservation. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

High Prevalence of Insecure Attachment in Patients with Primary Hypertension

PubMed Central

Balint, Elisabeth M.; Gander, Manuela; Pokorny, Dan; Funk, Alexandra; Waller, Christiane; Buchheim, Anna

2016-01-01

Hypertension is a major cardiovascular (CV) risk factor and is predicted by heightened CV reactivity to stress in healthy individuals. Patients with hypertension also show an altered stress response, while insecure attachment is linked to a heightened stress reactivity as well. This is the first study aiming to assess attachment representations in patients with primary hypertension and to investigate their CV responses when their attachment system is activated. We studied 50 patients (38 men, 12 women) with primary hypertension. The Adult Attachment Projective Picture System (AAP), a widely used and validated interview, was performed to measure the patients' attachment representations, and to activate their attachment system. Blood pressure and heart rate were measured after 10 min at rest prior to and directly after the AAP interview. Mood and state anxiety were assessed using the Multidimensional Mood State Questionnaire (MDBF) and the State Trait Anxiety Inventory-State (STAI-S) before and after the experiment. The prevalence of insecure attachment (dismissing, preoccupied, unresolved) in hypertensive patients was predominant (88%), while in non-clinical populations, only about 50% of individuals had insecure attachment patterns. Blood pressure (p < 0.001), heart rate (p = 0.016), and rate pressure product (p < 0.001) significantly increased in response to the attachment interview. Secure attached patients showed the highest rise in systolic blood pressure (p = 0.020) and the lowest heart rate compared to the other attachment groups (p = 0.043). However, attachment representation showed no significant group or interaction effects on diastolic blood pressure (DBP) and rate pressure product. Insecure attachment was highly over-represented in our sample of patients with primary hypertension. Additionally, a robust CV response to the attachment-activating stimulus was observed. Our data suggest that insecure attachment is significantly linked to primary hypertension

Interaction of Two Filaments in a Long Filament Channel Associated with Twin Coronal Mass Ejections

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zheng, Ruisheng; Chen, Yao; Wang, Bing

Using the high-quality observations of the Solar Dynamics Observatory , we present the interaction of two filaments (F1 and F2) in a long filament channel associated with twin coronal mass ejections (CMEs) on 2016 January 26. Before the eruption, a sequence of rapid cancellation and emergence of the magnetic flux has been observed, which likely triggered the ascending of the west filament (F1). The east footpoints of rising F1 moved toward the east far end of the filament channel, accompanied by post-eruption loops and flare ribbons. This likely indicated a large-scale eruption involving the long filament channel, which resulted frommore » the interaction between F1 and the east filament (F2). Some bright plasma flew over F2, and F2 stayed at rest during the eruption, likely due to the confinement of its overlying lower magnetic field. Interestingly, the impulsive F1 pushed its overlying magnetic arcades to form the first CME, and F1 finally evolved into the second CME after the collision with the nearby coronal hole. We suggest that the interaction of F1 and the overlying magnetic field of F2 led to the merging reconnection that forms a longer eruptive filament loop. Our results also provide a possible picture of the origin of twin CMEs and show that the large-scale magnetic topology of the coronal hole is important for the eventual propagation direction of CMEs.« less

Single-cell sequencing provides clues about the host interactions of segmented filamentous bacteria (SFB)

PubMed Central

Pamp, Sünje J.; Harrington, Eoghan D.; Quake, Stephen R.; Relman, David A.; Blainey, Paul C.

2012-01-01

Segmented filamentous bacteria (SFB) are host-specific intestinal symbionts that comprise a distinct clade within the Clostridiaceae, designated Candidatus Arthromitus. SFB display a unique life cycle within the host, involving differentiation into multiple cell types. The latter include filaments that attach intimately to intestinal epithelial cells, and from which “holdfasts” and spores develop. SFB induce a multifaceted immune response, leading to host protection from intestinal pathogens. Cultivation resistance has hindered characterization of these enigmatic bacteria. In the present study, we isolated five SFB filaments from a mouse using a microfluidic device equipped with laser tweezers, generated genome sequences from each, and compared these sequences with each other, as well as to recently published SFB genome sequences. Based on the resulting analyses, SFB appear to be dependent on the host for a variety of essential nutrients. SFB have a relatively high abundance of predicted proteins devoted to cell cycle control and to envelope biogenesis, and have a group of SFB-specific autolysins and a dynamin-like protein. Among the five filament genomes, an average of 8.6% of predicted proteins were novel, including a family of secreted SFB-specific proteins. Four ADP-ribosyltransferase (ADPRT) sequence types, and a myosin-cross-reactive antigen (MCRA) protein were discovered; we hypothesize that they are involved in modulation of host responses. The presence of polymorphisms among mouse SFB genomes suggests the evolution of distinct SFB lineages. Overall, our results reveal several aspects of SFB adaptation to the mammalian intestinal tract. PMID:22434425

Arabidopsis ribosomal proteins control vacuole trafficking and developmental programs through the regulation of lipid metabolism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Ruixi; Sun, Ruobai; Hicks, Glenn R.

The vacuole is the most prominent compartment in plant cells and is important for ion and protein storage. In our effort to search for key regulators in the plant vacuole sorting pathway, ribosomal large subunit 4 (rpl4d) was identified as a translational mutant defective in both vacuole trafficking and normal development. Polysome profiling of the rpl4d mutant showed reduction in polysome-bound mRNA compared with wild-type, but no significant change in the general mRNA distribution pattern. Ribsomal profiling data indicated that genes in the lipid metabolism pathways were translationally down-regulated in the rpl4d mutant. Live imaging studies by Nile red stainingmore » suggested that both polar and nonpolar lipid accumulation was reduced in meristem tissues of rpl4d mutants. Pharmacological evidence showed that sterol and sphingolipid biosynthetic inhibitors can phenocopy the defects of the rpl4d mutant, including an altered vacuole trafficking pattern. Genetic evidence from lipid biosynthetic mutants indicates that alteration in the metabolism of either sterol or sphingolipid biosynthesis resulted in vacuole trafficking defects, similar to the rpl4d mutant. Tissue-specific complementation with key enzymes from lipid biosynthesis pathways can partially rescue both vacuole trafficking and auxin-related developmental defects in the rpl4d mutant. These results indicate that lipid metabolism modulates auxin-mediated tissue differentiation and endomembrane trafficking pathways downstream of ribosomal protein function.« less

Arabidopsis ribosomal proteins control vacuole trafficking and developmental programs through the regulation of lipid metabolism

DOE PAGES

Li, Ruixi; Sun, Ruobai; Hicks, Glenn R.; ...

2014-12-22

The vacuole is the most prominent compartment in plant cells and is important for ion and protein storage. In our effort to search for key regulators in the plant vacuole sorting pathway, ribosomal large subunit 4 (rpl4d) was identified as a translational mutant defective in both vacuole trafficking and normal development. Polysome profiling of the rpl4d mutant showed reduction in polysome-bound mRNA compared with wild-type, but no significant change in the general mRNA distribution pattern. Ribsomal profiling data indicated that genes in the lipid metabolism pathways were translationally down-regulated in the rpl4d mutant. Live imaging studies by Nile red stainingmore » suggested that both polar and nonpolar lipid accumulation was reduced in meristem tissues of rpl4d mutants. Pharmacological evidence showed that sterol and sphingolipid biosynthetic inhibitors can phenocopy the defects of the rpl4d mutant, including an altered vacuole trafficking pattern. Genetic evidence from lipid biosynthetic mutants indicates that alteration in the metabolism of either sterol or sphingolipid biosynthesis resulted in vacuole trafficking defects, similar to the rpl4d mutant. Tissue-specific complementation with key enzymes from lipid biosynthesis pathways can partially rescue both vacuole trafficking and auxin-related developmental defects in the rpl4d mutant. These results indicate that lipid metabolism modulates auxin-mediated tissue differentiation and endomembrane trafficking pathways downstream of ribosomal protein function.« less

THE USE OF FORMALDEHYDE-TREATED 131I-ALBUMIN IN THE STUDY OF DIGESTIVE VACUOLES AND SOME PROPERTIES OF THESE PARTICLES FROM MOUSE LIVER

PubMed Central

Mego, John L.; Bertini, Francisco; McQueen, J. Donald

1967-01-01

The trichloroacetic acid-soluble radioactivity released during incubation of mouse liver particles containing intravenously injected formaldehyde-treated 131I-albumin consisted almost entirely of 131I-iodotyrosine. The material was shown to be excreted into the medium and was not due to disruption of the particles by acid. Triton X-100 or the absence of sucrose in the medium inhibited hydrolysis of the particle-associated labeled protein. This inhibition was due to disruption of the digestive vacuoles and dilution of the protein and cathepsins in the suspending medium. These results and other experimental evidence strongly suggest that the 131I-albumin-containing liver particles are digestive vacuoles. The results also establish that 131I-albumin may be used to study these vacuoles. High concentrations of sucrose (1 M) inhibited degradation of intraparticulate protein. However, 1 M salts inhibited only the rate of the digestion. Sucrose had an inhibitory effect on a crude cathepsin preparation, and salts stimulated the activity when 131I-albumin was used as substrate. The effect of high sucrose concentrations as an inhibitor of protein hydrolysis within digestive vacuoles was, therefore, most likely due principally to an inhibition of cathepsin activity within the vacuoles. The effect of salt was probably caused by a stimulation of both intra- and extra-particulate cathepsin activities, although 0.5–1.0 M KCl appeared to protect the particles. PMID:6034485

Characterizing filaments in regions of high-mass star formation: High-resolution submilimeter imaging of the massive star-forming complex NGC 6334 with ArTéMiS

NASA Astrophysics Data System (ADS)

André, Ph.; Revéret, V.; Könyves, V.; Arzoumanian, D.; Tigé, J.; Gallais, P.; Roussel, H.; Le Pennec, J.; Rodriguez, L.; Doumayrou, E.; Dubreuil, D.; Lortholary, M.; Martignac, J.; Talvard, M.; Delisle, C.; Visticot, F.; Dumaye, L.; De Breuck, C.; Shimajiri, Y.; Motte, F.; Bontemps, S.; Hennemann, M.; Zavagno, A.; Russeil, D.; Schneider, N.; Palmeirim, P.; Peretto, N.; Hill, T.; Minier, V.; Roy, A.; Rygl, K. L. J.

2016-07-01

Context. Herschel observations of nearby molecular clouds suggest that interstellar filaments and prestellar cores represent two fundamental steps in the star formation process. The observations support a picture of low-mass star formation according to which filaments of ~0.1 pc width form first in the cold interstellar medium, probably as a result of large-scale compression of interstellar matter by supersonic turbulent flows, and then prestellar cores arise from gravitational fragmentation of the densest filaments. Whether this scenario also applies to regions of high-mass star formation is an open question, in part because the resolution of Herschel is insufficient to resolve the inner width of filaments in the nearest regions of massive star formation. Aims: In an effort to characterize the inner width of filaments in high-mass star-forming regions, we imaged the central part of the NGC 6334 complex at a resolution higher by a factor of >3 than Herschel at 350 μm. Methods: We used the large-format bolometer camera ArTéMiS on the APEX telescope and combined the high-resolution ArTéMiS data at 350 μm with Herschel/HOBYS data at 70-500 μm to ensure good sensitivity to a broad range of spatial scales. This allowed us to study the structure of the main narrow filament of the complex with a resolution of 8″ or <0.07 pc at d ~ 1.7 kpc. Results: Our study confirms that this filament is a very dense, massive linear structure with a line mass ranging from ~500 M⊙/pc to ~2000 M⊙/pc over nearly 10 pc. It also demonstrates for the first time that its inner width remains as narrow as W ~ 0.15 ± 0.05 pc all along the filament length, within a factor of <2 of the characteristic 0.1 pc value found with Herschel for lower-mass filaments in the Gould Belt. Conclusions: While it is not completely clear whether the NGC 6334 filament will form massive stars in the future, it is two to three orders of magnitude denser than the majority of filaments observed in Gould Belt

Identification of a Peptide-Pheromone that Enhances Listeria monocytogenes Escape from Host Cell Vacuoles

PubMed Central

Xayarath, Bobbi; Alonzo, Francis; Freitag, Nancy E.

2015-01-01

Listeria monocytogenes is a Gram-positive facultative intracellular bacterial pathogen that invades mammalian cells and escapes from membrane-bound vacuoles to replicate within the host cell cytosol. Gene products required for intracellular bacterial growth and bacterial spread to adjacent cells are regulated by a transcriptional activator known as PrfA. PrfA becomes activated following L. monocytogenes entry into host cells, however the signal that stimulates PrfA activation has not yet been defined. Here we provide evidence for L. monocytogenes secretion of a small peptide pheromone, pPplA, which enhances the escape of L. monocytogenes from host cell vacuoles and may facilitate PrfA activation. The pPplA pheromone is generated via the proteolytic processing of the PplA lipoprotein secretion signal peptide. While the PplA lipoprotein is dispensable for pathogenesis, bacteria lacking the pPplA pheromone are significantly attenuated for virulence in mice and have a reduced efficiency of bacterial escape from the vacuoles of nonprofessional phagocytic cells. Mutational activation of PrfA restores virulence and eliminates the need for pPplA-dependent signaling. Experimental evidence suggests that the pPplA peptide may help signal to L. monocytogenes its presence within the confines of the host cell vacuole, stimulating the expression of gene products that contribute to vacuole escape and facilitating PrfA activation to promote bacterial growth within the cytosol. PMID:25822753

Powering of Hα Filaments by Cosmic Rays

NASA Astrophysics Data System (ADS)

Ruszkowski, Mateusz; Yang, H.-Y. Karen; Reynolds, Christopher S.

2018-05-01

Cluster cool cores possess networks of line-emitting filaments. These filaments are thought to originate via uplift of cold gas from cluster centers by buoyant active galactic nuclei (AGNs) bubbles, or via local thermal instability in the hot intracluster medium (ICM). Therefore, the filaments are either the signatures of AGN feedback or feeding of supermassive black holes. Despite being characterized by very short cooling times, the filaments are significant Hα emitters, which suggests that some process continuously powers these structures. Many cool cores host diffuse radio mini halos and AGN injecting radio plasma, suggesting that cosmic rays (CRs) and magnetic fields are present in the ICM. We argue that the excitation of Alfvén waves by CR streaming, and the replenishment of CR energy via accretion onto the filaments of high-plasma-β ICM characterized by low CR pressure support, can provide the adequate amount of heating to power and sustain the emission from these filaments. This mechanism does not require the CRs to penetrate the filaments, even if the filaments are magnetically isolated from the ambient ICM, and it may operate irrespectively of whether the filaments are dredged up from the center or form in situ in the ICM. This picture is qualitatively consistent with non-thermal line ratios seen in the cold filaments. Future X-ray observations of the iron line complex with XARM, Lynx, or Athena could help to test this model by providing constraints on the amount of CRs in the hot plasma that is cooling and accreting onto the filaments.

Aerogel-supported filament

DOEpatents

Wuest, C.R.; Tillotson, T.M.; Johnson, C.V. III

1995-05-16

The present invention is a thin filament embedded in a low density aerogel for use in radiation detection instruments and incandescent lamps. The aerogel provides a supportive matrix that is thermally and electrically nonconductive, mechanically strong, highly porous, gas-permeable, and transparent to ionizing radiation over short distances. A low density, open-cell aerogel is cast around a fine filament or wire, which allows the wire to be positioned with little or no tension and keeps the wire in place in the event of breakage. The aerogel support reduces the stresses on the wire caused by vibrational, gravitational, electrical, and mechanical forces. 6 Figs.

Thermal design of composite material high temperature attachments

NASA Technical Reports Server (NTRS)

1972-01-01

An evaluation has been made of the thermal aspects of utilizing advanced filamentary composite materials as primary structures on the shuttle vehicle. The technical objectives of this study are to: (1) establish and design concepts for maintaining material temperatures within allowable limits at TPS attachments and or penetrations applicable to the space shuttle; and (2) verify the thermal design analysis by testing selected concepts. Specific composite materials being evaluated are boron epoxy, graphite/epoxy, boron polyimide, and boron aluminum; graphite/polyimide has been added to this list for property data identification and preliminary evaluation of thermal design problems. The TPS standoff to composite structure attachment over-temperature problem is directly related to TPS maximum surface temperature. To provide a thermally comprehensive evaluation of attachment temperature characteristics, maximum surface temperatures of 900 F, 1200 F, 1800 F, 2500 F and 3000 F are considered in this study. This range of surface temperatures and the high and low maximum temperature capability of the selected composite materials will result in a wide range of thermal requirements for composite/TPS standoff attachments.

The Identification of Cable Bacteria Attached to the Anode of a Benthic Microbial Fuel Cell: Evidence of Long Distance Extracellular Electron Transport to Electrodes

PubMed Central

Reimers, Clare E.; Li, Cheng; Graw, Michael F.; Schrader, Paul S.; Wolf, Michael

2017-01-01

Multicellular, filamentous, sulfur-oxidizing bacteria, known as cable bacteria, were discovered attached to fibers of a carbon brush electrode serving as an anode of a benthic microbial fuel cell (BMFC). The BMFC had been operated in a temperate estuarine environment for over a year before collecting anode samples for scanning electron microscopy and phylogenetic analyses. Individual filaments were attached by single terminus cells with networks of pilus-like nano-filaments radiating out from these cells, across the anode fiber surface, and between adjacent attachment locations. Current harvesting by the BMFC poised the anode at potentials of ~170–250 mV vs. SHE, and these surface potentials appear to have allowed the cable bacteria to use the anode as an electron acceptor in a completely anaerobic environment. A combination of catalyzed reporter deposition fluorescent in situ hybridization (CARD-FISH) and 16S rRNA gene sequence analysis confirmed the phylogeny of the cable bacteria and showed that filaments often occurred in bundles and in close association with members of the genera Desulfuromonas. However, the Desulfobulbaceae Operational Taxonomic Units (OTUs) from the 16S sequencing did not cluster closely with other putative cable bacteria sequences suggesting that the taxonomic delineation of cable bacteria is far from complete. PMID:29114243

Leishmania amazonensis Engages CD36 to Drive Parasitophorous Vacuole Maturation

PubMed Central

Okuda, Kendi; Tong, Mei; Dempsey, Brian; Moore, Kathryn J.; Gazzinelli, Ricardo T.; Silverman, Neal

2016-01-01

Leishmania amastigotes manipulate the activity of macrophages to favor their own success. However, very little is known about the role of innate recognition and signaling triggered by amastigotes in this host-parasite interaction. In this work we developed a new infection model in adult Drosophila to take advantage of its superior genetic resources to identify novel host factors limiting Leishmania amazonensis infection. The model is based on the capacity of macrophage-like cells, plasmatocytes, to phagocytose and control the proliferation of parasites injected into adult flies. Using this model, we screened a collection of RNAi-expressing flies for anti-Leishmania defense factors. Notably, we found three CD36-like scavenger receptors that were important for defending against Leishmania infection. Mechanistic studies in mouse macrophages showed that CD36 accumulates specifically at sites where the parasite contacts the parasitophorous vacuole membrane. Furthermore, CD36-deficient macrophages were defective in the formation of the large parasitophorous vacuole typical of L. amazonensis infection, a phenotype caused by inefficient fusion with late endosomes and/or lysosomes. These data identify an unprecedented role for CD36 in the biogenesis of the parasitophorous vacuole and further highlight the utility of Drosophila as a model system for dissecting innate immune responses to infection. PMID:27280707

Characterization of a filamentous biofilm community established in a cellulose-fed microbial fuel cell.

PubMed

Ishii, Shun'ichi; Shimoyama, Takefumi; Hotta, Yasuaki; Watanabe, Kazuya

2008-01-10

Microbial fuel cells (MFCs) are devices that exploit microorganisms to generate electric power from organic matter. Despite the development of efficient MFC reactors, the microbiology of electricity generation remains to be sufficiently understood. A laboratory-scale two-chamber microbial fuel cell (MFC) was inoculated with rice paddy field soil and fed cellulose as the carbon and energy source. Electricity-generating microorganisms were enriched by subculturing biofilms that attached onto anode electrodes. An electric current of 0.2 mA was generated from the first enrichment culture, and ratios of the major metabolites (e.g., electric current, methane and acetate) became stable after the forth enrichment. In order to investigate the electrogenic microbial community in the anode biofilm, it was morphologically analyzed by electron microscopy, and community members were phylogenetically identified by 16S rRNA gene clone-library analyses. Electron microscopy revealed that filamentous cells and rod-shaped cells with prosthecae-like filamentous appendages were abundantly present in the biofilm. Filamentous cells and appendages were interconnected via thin filaments. The clone library analyses frequently detected phylotypes affiliated with Clostridiales, Chloroflexi, Rhizobiales and Methanobacterium. Fluorescence in-situ hybridization revealed that the Rhizobiales population represented rod-shaped cells with filamentous appendages and constituted over 30% of the total population. Bacteria affiliated with the Rhizobiales constituted the major population in the cellulose-fed MFC and exhibited unique morphology with filamentous appendages. They are considered to play important roles in the cellulose-degrading electrogenic community.

Characterization of a filamentous biofilm community established in a cellulose-fed microbial fuel cell

PubMed Central

Ishii, Shun'ichi; Shimoyama, Takefumi; Hotta, Yasuaki; Watanabe, Kazuya

2008-01-01

Background Microbial fuel cells (MFCs) are devices that exploit microorganisms to generate electric power from organic matter. Despite the development of efficient MFC reactors, the microbiology of electricity generation remains to be sufficiently understood. Results A laboratory-scale two-chamber microbial fuel cell (MFC) was inoculated with rice paddy field soil and fed cellulose as the carbon and energy source. Electricity-generating microorganisms were enriched by subculturing biofilms that attached onto anode electrodes. An electric current of 0.2 mA was generated from the first enrichment culture, and ratios of the major metabolites (e.g., electric current, methane and acetate) became stable after the forth enrichment. In order to investigate the electrogenic microbial community in the anode biofilm, it was morphologically analyzed by electron microscopy, and community members were phylogenetically identified by 16S rRNA gene clone-library analyses. Electron microscopy revealed that filamentous cells and rod-shaped cells with prosthecae-like filamentous appendages were abundantly present in the biofilm. Filamentous cells and appendages were interconnected via thin filaments. The clone library analyses frequently detected phylotypes affiliated with Clostridiales, Chloroflexi, Rhizobiales and Methanobacterium. Fluorescence in-situ hybridization revealed that the Rhizobiales population represented rod-shaped cells with filamentous appendages and constituted over 30% of the total population. Conclusion Bacteria affiliated with the Rhizobiales constituted the major population in the cellulose-fed MFC and exhibited unique morphology with filamentous appendages. They are considered to play important roles in the cellulose-degrading electrogenic community. PMID:18186940

Extreme Nonlinear Optics of High Intensity Laser Pulse Filamentation in Gases

DTIC Science & Technology

2016-05-12

of energy from femtosecond filaments. Published * absolute measurements of electronic, vibrational, and rotational nonlinear response in H2 and D2 ...coefficients in the fastest rotating molecules H2 and D2 , which can serve as a benchmark for theory of high field molecule interactions. One of the...17. Absolute measurement of the ultrafast nonlinear electronic and rovibrational response in H2 and D2 J. K. Wahlstrand, S. Zahedpour, Y.-H

Phenylpropanoid Scent Compounds in Petunia x hybrida Are Glycosylated and Accumulate in Vacuoles

PubMed Central

Cna'ani, Alon; Shavit, Reut; Ravid, Jasmin; Aravena-Calvo, Javiera; Skaliter, Oded; Masci, Tania; Vainstein, Alexander

2017-01-01

Floral scent has been studied extensively in the model plant Petunia. However, little is known about the intracellular fate of scent compounds. Here, we characterize the glycosylation of phenylpropanoid scent compounds in Petunia x hybrida. This modification reduces scent compounds' volatility, reactivity, and autotoxicity while increasing their water-solubility. Gas chromatography–mass spectrometry (GC–MS) analyses revealed that flowers of petunia cultivars accumulate substantial amounts of glycosylated scent compounds and that their increasing level parallels flower development. In contrast to the pool of accumulated aglycones, which drops considerably at the beginning of the light period, the collective pool of glycosides starts to increase at that time and does not decrease thereafter. The glycoside pool is dynamic and is generated or catabolized during peak scent emission, as inferred from phenylalanine isotope-feeding experiments. Using several approaches, we show that phenylpropanoid scent compounds are stored as glycosides in the vacuoles of petal cells: ectopic expression of Aspergillus niger β-glucosidase-1 targeted to the vacuole resulted in decreased glycoside accumulation; GC–MS analysis of intact vacuoles isolated from petal protoplasts revealed the presence of glycosylated scent compounds. Accumulation of glycosides in the vacuoles seems to be a common mechanism for phenylpropanoid metabolites. PMID:29163617

System Applies Polymer Powder To Filament Tow

NASA Technical Reports Server (NTRS)

Baucom, Robert M.; Snoha, John J.; Marchello, Joseph M.

1993-01-01

Polymer powder applied uniformly and in continuous manner. Powder-coating system applies dry polymer powder to continuous fiber tow. Unique filament-spreading technique, combined with precise control of tension on fibers in system, ensures uniform application of polymer powder to web of spread filaments. Fiber tows impregnated with dry polymer powders ("towpregs") produced for preform-weaving and composite-material-molding applications. System and process valuable to prepreg industry, for production of flexible filament-windable tows and high-temperature polymer prepregs.

Microwave metamaterials made by fused deposition 3D printing of a highly conductive copper-based filament

NASA Astrophysics Data System (ADS)

Xie, Yangbo; Ye, Shengrong; Reyes, Christopher; Sithikong, Pariya; Popa, Bogdan-Ioan; Wiley, Benjamin J.; Cummer, Steven A.

2017-05-01

This work reports a method for fabricating three-dimensional microwave metamaterials by fused deposition modeling 3D printing of a highly conductive polymer composite filament. The conductivity of such a filament is shown to be nearly equivalent to that of a perfect conductor for microwave metamaterial applications. The expanded degrees-of-freedom made available by 3D metamaterial designs are demonstrated by designing, fabricating, and testing a 3D-printed unit cell with a broadband permittivity as high as 14.4. The measured and simulated S-parameters agree well with a mean squared error smaller than 0.1. The presented method not only allows reliable and convenient fabrication of microwave metamaterials with high conductivity but also opens the door to exploiting the third dimension of the unit cell design space to achieve enhanced electromagnetic properties.

Cryo-EM reconstruction of AlfA from Bacillus subtilis reveals the structure of a simplified actin-like filament at 3.4-Å resolution.

PubMed

Szewczak-Harris, Andrzej; Löwe, Jan

2018-03-27

Low copy-number plasmid pLS32 of Bacillus subtilis subsp. natto contains a partitioning system that ensures segregation of plasmid copies during cell division. The partitioning locus comprises actin-like protein AlfA, adaptor protein AlfB, and the centromeric sequence parN Similar to the ParMRC partitioning system from Escherichia coli plasmid R1, AlfA filaments form actin-like double helical filaments that arrange into an antiparallel bipolar spindle, which attaches its growing ends to sister plasmids through interactions with AlfB and parN Because, compared with ParM and other actin-like proteins, AlfA is highly diverged in sequence, we determined the atomic structure of nonbundling AlfA filaments to 3.4-Å resolution by cryo-EM. The structure reveals how the deletion of subdomain IIB of the canonical actin fold has been accommodated by unique longitudinal and lateral contacts, while still enabling formation of left-handed, double helical, polar and staggered filaments that are architecturally similar to ParM. Through cryo-EM reconstruction of bundling AlfA filaments, we obtained a pseudoatomic model of AlfA doublets: the assembly of two filaments. The filaments are antiparallel, as required by the segregation mechanism, and exactly antiphasic with near eightfold helical symmetry, to enable efficient doublet formation. The structure of AlfA filaments and doublets shows, in atomic detail, how deletion of an entire domain of the actin fold is compensated by changes to all interfaces so that the required properties of polymerization, nucleotide hydrolysis, and antiparallel doublet formation are retained to fulfill the system's biological raison d'être.

A penny-shaped crack in a filament reinforced matrix. 1: The filament model

NASA Technical Reports Server (NTRS)

Erdogan, F.; Pacella, A. H.

1973-01-01

The electrostatic problem of a penny-shaped crack in an elastic matrix which reinforced by filaments or fibers perpendicular to the plane of the crack was studied. The elastic filament model was developed for application to evaluation studies of the stress intensity factor along the periphery of the crack, the stresses in the filaments or fibers, and the interface shear between the matrix and the filaments or fibers. The requirements expected of the model are a sufficiently accurate representation of the filament and applicability to the interaction problems involving a cracked elastic continuum with multi-filament reinforcements. The technique for developing the model and numerical examples of it are shown.

Beam wandering of femtosecond laser filament in air.

PubMed

Yang, Jing; Zeng, Tao; Lin, Lie; Liu, Weiwei

2015-10-05

The spatial wandering of a femtosecond laser filament caused by the filament heating effect in air has been studied. An empirical formula has also been derived from the classical Karman turbulence model, which determines quantitatively the displacement of the beam center as a function of the propagation distance and the effective turbulence structure constant. After fitting the experimental data with this formula, the effective turbulence structure constant has been estimated for a single filament generated in laboratory environment. With this result, one may be able to estimate quantitatively the displacement of a filament over long distance propagation and interpret the practical performance of the experiments assisted by femtosecond laser filamentation, such as remote air lasing, pulse compression, high order harmonic generation (HHG), etc.

Probing the contractile vacuole as Achilles' heel of the biotrophic grapevine pathogen Plasmopara viticola.

PubMed

Tröster, Viktoria; Setzer, Tabea; Hirth, Thomas; Pecina, Anna; Kortekamp, Andreas; Nick, Peter

2017-09-01

The causative agent of Grapevine Downy Mildew, the oomycete Plasmopara viticola, poses a serious threat to viticulture. In the current work, the contractile vacuole of the zoospore is analysed as potential target for novel plant protection strategies. Using a combination of electron microscopy, spinning disc confocal microscopy, and video differential interference contrast microscopy, we have followed the genesis and dynamics of this vacuole required during the search for the stomata, when the non-walled zoospore is exposed to hypotonic conditions. This subcellular description was combined with a pharmacological study, where the functionality of the contractile vacuole was blocked by manipulation of actin, by Na, Cu, and Al ions or by inhibition of the NADPH oxidase. We further observe that RGD peptides (mimicking binding sites for integrins at the extracellular matrix) can inhibit the function of the contractile vacuole as well. Finally, we show that an extract from Chinese liquorice (Glycyrrhiza uralensis) proposed as biocontrol for Downy Mildews can efficiently induce zoospore burst and that this activity depends on the activity of NADPH oxidase. The effect of the extract can be phenocopied by its major compound, glycyrrhizin, suggesting a mode of action for this biologically safe alternative to copper products.

HV discharge acceleration by sequences of UV laser filaments with visible and near-infrared pulses

NASA Astrophysics Data System (ADS)

Schubert, Elise; Rastegari, Ali; Feng, Chengyong; Mongin, Denis; Kamer, Brian; Kasparian, Jérôme; Wolf, Jean-Pierre; Arissian, Ladan; Diels, Jean-Claude

2017-12-01

We investigate the triggering and guiding of DC high-voltage discharges over a distance of 37 cm by filaments produced by ultraviolet (266 nm) laser pulses of 200 ps duration. The latter reduce the breakdown electric field by half and allow up to 80% discharge probability in an electric field of 920 kV m–1. This high efficiency is not further increased by adding nanosecond pulses in the Joule range at 532 and at 1064 nm. However, the latter statistically increases the guiding length, thereby accelerating the discharge by a factor of 2. This effect is due both to photodetachment and to the heating of the plasma channel, that increases the efficiency of avalanche ionization and reduces electron attachment and recombination.

From Aβ Filament to Fibril: Molecular Mechanism of Surface-Activated Secondary Nucleation from All-Atom MD Simulations.

PubMed

Schwierz, Nadine; Frost, Christina V; Geissler, Phillip L; Zacharias, Martin

2017-02-02

Secondary nucleation pathways in which existing amyloid fibrils catalyze the formation of new aggregates and neurotoxic oligomers are of immediate importance for the onset and progression of Alzheimer's disease. Here, we apply extensive all-atom molecular dynamics simulations in explicit water to study surface-activated secondary nucleation pathways at the extended lateral β-sheet surface of a preformed Aβ 9-40 filament. Calculation of free-energy profiles allows us to determine binding free energies and conformational intermediates for nucleation complexes consisting of 1-4 Aβ peptides. In addition, we combine the free-energy profiles with position-dependent diffusion profiles to extract complementary kinetic information and macroscopic growth rates. Single monomers bind to the β-sheet surface in a disordered, hydrophobically collapsed conformation, whereas dimers and larger oligomers can retain a cross-β conformation resembling a more ordered fibril structure. The association processes during secondary nucleation follow a dock/lock mechanism consisting of a fast initial encounter phase (docking) and a slow structural rearrangement phase (locking). The major driving forces for surface-activated secondary nucleation are the release of a large number of hydration water molecules and the formation of hydrophobic interface contacts, the latter being in contrast to the elongation process at filament tips, which is dominated by the formation of stable and highly specific interface hydrogen bonds. The calculated binding free energies and the association rates for the attachment of Aβ monomers and oligomers to the extended lateral β-sheet surface of the filament seed are higher compared to those for elongation at the filament tips, indicating that secondary nucleation pathways can become important once a critical concentration of filaments has formed.

High Current, Multi-Filament Photoconductive Semiconductor Switching

DTIC Science & Technology

2011-06-01

linear PCSS triggered with a 100 fs laser pulse . Figure 1. A generic photoconductive semiconductor switch rapidly discharges a charged capacitor...switching is the most critical challenge remaining for photoconductive semiconductor switch (PCSS) applications in Pulsed Power. Many authors have...isolation and control, pulsed or DC charging, and long device lifetime, provided the current per filament is limited to 20-30A for short pulse (10

Filamentary structures in dense plasma focus: Current filaments or vortex filaments?

DOE Office of Scientific and Technical Information (OSTI.GOV)

Soto, Leopoldo, E-mail: lsoto@cchen.cl; Pavez, Cristian; Moreno, José

2014-07-15

Recent observations of an azimuthally distributed array of sub-millimeter size sources of fusion protons and correlation between extreme ultraviolet (XUV) images of filaments with neutron yield in PF-1000 plasma focus have re-kindled interest in their significance. These filaments have been described variously in literature as current filaments and vortex filaments, with very little experimental evidence in support of either nomenclature. This paper provides, for the first time, experimental observations of filaments on a table-top plasma focus device using three techniques: framing photography of visible self-luminosity from the plasma, schlieren photography, and interferometry. Quantitative evaluation of density profile of filaments frommore » interferometry reveals that their radius closely agrees with the collision-less ion skin depth. This is a signature of relaxed state of a Hall fluid, which has significant mass flow with equipartition between kinetic and magnetic energy, supporting the “vortex filament” description. This interpretation is consistent with empirical evidence of an efficient energy concentration mechanism inferred from nuclear reaction yields.« less

Glial S100B Positive Vacuoles In Purkinje Cells: Earliest Morphological Abnormality In SCA1 Transgenic Mice

PubMed Central

VIG, Parminder J.S.; LOPEZ, Maripar E.; WEI, Jinrong; D’SOUZA, David R.; SUBRAMONY, SH; HENEGAR, Jeffrey; FRATKIN, Jonathan D.

2007-01-01

Spinocerebellar ataxia-1 (SCA1) is caused by the expansion of a polyglutamine repeat within the disease protein, ataxin-1. The overexpression of mutant ataxin-1 in SCA1 transgenic mice results in the formation of cytoplasmic vacuoles in Purkinje neurons (PKN) of the cerebellum. PKN are closely associated with neighboring Bergmann glia. To elucidate the role of Bergmann glia in SCA1 pathogenesis, cerebellar tissue from 7 days to 6 wks old SCA1 transgenic and wildtype mice were used. We observed that Bergmann glial S100B protein is localized to the cytoplasmic vacuoles in SCA1 PKN. These S100B positive cytoplasmic vacuoles began appearing much before the onset of behavioral abnormalities, and were negative for other glial and PKN marker proteins. Electron micrographs revealed that vacuoles have a double membrane. In the vacuoles, S100B colocalized with receptors of advanced glycation end-products (RAGE), and S100B co-immunoprecipated with cerebellar RAGE. In SCA1 PKN cultures, exogenous S100B protein interacted with the PKN membranes and was internalized. These data suggest that glial S100B though extrinsic to PKN is sequestered into cytoplasmic vacuoles in SCA1 mice at early postnatal ages. Further, S100B may be binding to RAGE on Purkinje cell membranes before these membranes are internalized. PMID:18176630

High-resolution electron microscopic evidence for the filamentous structure of the cyst wall in Giardia muris and Giardia duodenalis.

PubMed

Erlandsen, S L; Bemrick, W J; Pawley, J

1989-10-01

High-resolution morphological studies of the cyst wall of Giardia spp. were performed using low-voltage scanning electron microscopy (LVSEM) and transmission electron microscopy (TEM). The cyst wall was composed of membranous and filamentous layers. The membranous layer consisted of an inner and an outer cyst membrane separated by a thin layer of cytoplasm. The filamentous layer contained individual filaments that ranged from 7 to 20 nm in diameter when measured by LVSEM, formed a dense meshwork with branches or interconnections, and were occasionally arranged on the surface in whorled patterns. Cysts of Giardia muris from mice, Giardia duodenalis from dogs, pigs, voles, beavers, muskrats, and humans, and Giardia psittaci from a bird (parakeet), possessed an essentially identical wall composed of filaments. Inducement of excystation in viable Giardia cysts produced a dramatic increase in the interfilament spacing over an entire cyst, but none was observed in heat-killed or chemically fixed control cysts. These results demonstrated that the cyst wall of Giardia spp. was composed of a complex arrangement of filaments, presumably formed during the process of encystment.

Light sources based on semiconductor current filaments

DOEpatents

Zutavern, Fred J.; Loubriel, Guillermo M.; Buttram, Malcolm T.; Mar, Alan; Helgeson, Wesley D.; O'Malley, Martin W.; Hjalmarson, Harold P.; Baca, Albert G.; Chow, Weng W.; Vawter, G. Allen

2003-01-01

The present invention provides a new type of semiconductor light source that can produce a high peak power output and is not injection, e-beam, or optically pumped. The present invention is capable of producing high quality coherent or incoherent optical emission. The present invention is based on current filaments, unlike conventional semiconductor lasers that are based on p-n junctions. The present invention provides a light source formed by an electron-hole plasma inside a current filament. The electron-hole plasma can be several hundred microns in diameter and several centimeters long. A current filament can be initiated optically or with an e-beam, but can be pumped electrically across a large insulating region. A current filament can be produced in high gain photoconductive semiconductor switches. The light source provided by the present invention has a potentially large volume and therefore a potentially large energy per pulse or peak power available from a single (coherent) semiconductor laser. Like other semiconductor lasers, these light sources will emit radiation at the wavelength near the bandgap energy (for GaAs 875 nm or near infra red). Immediate potential applications of the present invention include high energy, short pulse, compact, low cost lasers and other incoherent light sources.

Two-dimensional photoacoustic imaging of femtosecond filament in water

NASA Astrophysics Data System (ADS)

Potemkin, F. V.; Mareev, E. I.; Rumiantsev, B. V.; Bychkov, A. S.; Karabutov, A. A.; Cherepetskaya, E. B.; Makarov, V. A.

2018-07-01

We report a first-of-its-kind optoacoustic tomography of a femtosecond filament in water. Using a broadband (~100 MHz) piezoelectric transducer and a back-projection reconstruction technique, a single filament profile was retrieved. Obtained pressure distribution induced by the femtosecond filament allowed us to identify the size of the core and the energy reservoir with spatial resolution better than 10 µm. The photoacoustic imaging provides direct measurements of the energy deposition into the medium under filamentation of ultrashort laser pulses that cannot be obtained by existing techniques. In combination with a relative simplicity and high accuracy, photoacoustic imaging can be considered as a breakthrough instrument for filamentation investigation.

Content and Vacuole/Extravacuole Distribution of Neutral Sugars, Free Amino Acids, and Anthocyanin in Protoplasts 1

PubMed Central

Wagner, George J.

1979-01-01

Neutral sugar, free amino acid, and anthocyanin levels and vacuole/extravacuole distribution were determined for Hippeastrum and Tulipa petal and Tulipa leaf protoplasts. Glucose and fructose, the predominant neutral monosaccharides observed, were primarily vacuolar in location. Glutamine, the predominant free amino acid found, was primarily extravacuolar. γ-Methyleneglutamate was identified as a major constituent of Tulipa protoplasts. Qualitative characterization of Hippeastrum petal and vacuole organic acids indicated the presence of oxalic, malic, citric, and isocitric acids. Data are presented which indicate that vacuoles obtained by gentle osmotic shock of protoplasts in dibasic phosphate have good purity and retain their contents. Images PMID:16660921

Sliding movement of single actin filaments on one-headed myosin filaments

NASA Astrophysics Data System (ADS)

Harada, Yoshie; Noguchi, Akira; Kishino, Akiyoshi; Yanagida, Toshio

1987-04-01

The myosin molecule consists of two heads, each of which contains an enzymatic active site and an actin-binding site. The fundamental problem of whether the two heads function independently or cooperatively during muscle contraction has been studied by methods using an actomyosin thread1, superprecipitation2-4 and chemical modification of muscle fibres5. No clear conclusion has yet been reached. We have approached this question using an assay system in which sliding movements of fluorescently labelled single actin filaments along myosin filaments can be observed directly6,7. Here, we report direct measurement of the sliding of single actin filaments along one-headed myosin filaments in which the density of heads was varied over a wide range. Our results show that cooperative interaction between the two heads of myosin is not essential for inducing the sliding movement of actin filaments.

Optical spectroscopy using gas-phase femtosecond laser filamentation.

PubMed

Odhner, Johanan; Levis, Robert

2014-01-01

Femtosecond laser filamentation occurs as a dynamic balance between the self-focusing and plasma defocusing of a laser pulse to produce ultrashort radiation as brief as a few optical cycles. This unique source has many properties that make it attractive as a nonlinear optical tool for spectroscopy, such as propagation at high intensities over extended distances, self-shortening, white-light generation, and the formation of an underdense plasma. The plasma channel that constitutes a single filament and whose position in space can be controlled by its input parameters can span meters-long distances, whereas multifilamentation of a laser beam can be sustained up to hundreds of meters in the atmosphere. In this review, we briefly summarize the current understanding and use of laser filaments for spectroscopic investigations of molecules. A theoretical framework of filamentation is presented, along with recent experimental evidence supporting the established understanding of filamentation. Investigations carried out on vibrational and rotational spectroscopy, filament-induced breakdown, fluorescence spectroscopy, and backward lasing are discussed.

Dramatic enhancement of supercontinuum generation in elliptically-polarized laser filaments

PubMed Central

Rostami, Shermineh; Chini, Michael; Lim, Khan; Palastro, John P.; Durand, Magali; Diels, Jean-Claude; Arissian, Ladan; Baudelet, Matthieu; Richardson, Martin

2016-01-01

Broadband laser sources based on supercontinuum generation in femtosecond laser filamentation have enabled applications from stand-off sensing and spectroscopy to the generation and self-compression of high-energy few-cycle pulses. Filamentation relies on the dynamic balance between self-focusing and plasma defocusing – mediated by the Kerr nonlinearity and multiphoton or tunnel ionization, respectively. The filament properties, including the supercontinuum generation, are therefore highly sensitive to the properties of both the laser source and the propagation medium. Here, we report the anomalous spectral broadening of the supercontinuum for filamentation in molecular gases, which is observed for specific elliptical polarization states of the input laser pulse. The resulting spectrum is accompanied by a modification of the supercontinuum polarization state and a lengthening of the filament plasma column. Our experimental results and accompanying simulations suggest that rotational dynamics of diatomic molecules play an essential role in filamentation-induced supercontinuum generation, which can be controlled with polarization ellipticity. PMID:26847427

Culture supernatants from V. cholerae O1 El Tor strains isolated from different geographic areas induce cell vacuolation and cytotoxicity.

PubMed

Vidal, Jorge E; Enríquez-Rincón, Fernando; Giono-Cerezo, Silvia; Ribas-Aparicio, Rosa María; Figueroa-Arredondo, Paula

2009-01-01

To investigate whether the HlyA-induced vacuolating effect is produced by V. cholerae O1 ElTor strains isolated from different geographic origins, including Mexico. Supernatant-induced haemolysis, vacuolating activity and cytotoxicity in Vero cells were recorded. PCR, RFLP analysis and molecular cloning were performed. All ElTor strains analyzed induced cellular vacuolation. Ribotype 2 strains isolates from the U.S. gulf coast yielded the highest titer of vacuolating activity. Eight of nine strains were haemolytic, while all strains were PCR positive for the hlyA gene. We cloned the hlyA gene from two ElTor strains, a toxigenic (2514-88, ctxAB+) and a non-toxigenic Mexican strain (CM 91-3, ctxAB-). Supernatant from those recombinant E. coli strains induced haemolysis, cell vacuolation and cytotoxicity. RFLP-PCR analysis revealed similarities in the hlyA gene from all strains tested. The HlyA-induced vacuolating effect is a widespread phenotype of epidemic V. cholerae O1 ElTor strains.

Fused Filament Fabrication of Prosthetic Components for Trans-Humeral Upper Limb Prosthetics

NASA Astrophysics Data System (ADS)

Lathers, Steven M.

Presented below is the design and fabrication of prosthetic components consisting of an attachment, tactile sensing, and actuator systems with Fused Filament Fabrication (FFF) technique. The attachment system is a thermoplastic osseointegrated upper limb prosthesis for average adult trans-humeral amputation with mechanical properties greater than upper limb skeletal bone. The prosthetic designed has: a one-step surgical process, large cavities for bone tissue ingrowth, uses a material that has an elastic modulus less than skeletal bone, and can be fabricated on one system. FFF osseointegration screw is an improvement upon the current two-part osseointegrated prosthetics that are composed of a fixture and abutment. The current prosthetic design requires two invasive surgeries for implantation and are made of titanium, which has an elastic modulus greater than bone. An elastic modulus greater than bone causes stress shielding and overtime can cause loosening of the prosthetic. The tactile sensor is a thermoplastic piezo-resistive sensor for daily activities for a prosthetic's feedback system. The tactile sensor is manufactured from a low elastic modulus composite comprising of a compressible thermoplastic elastomer and conductive carbon. Carbon is in graphite form and added in high filler ratios. The printed sensors were compared to sensors that were fabricated in a gravity mold to highlight the difference in FFF sensors to molded sensors. The 3D printed tactile sensor has a thickness and feel similar to human skin, has a simple fabrication technique, can detect forces needed for daily activities, and can be manufactured in to user specific geometries. Lastly, a biomimicking skeletal muscle actuator for prosthetics was developed. The actuator developed is manufactured with Fuse Filament Fabrication using a shape memory polymer composite that has non-linear contractile and passive forces, contractile forces and strains comparable to mammalian skeletal muscle, reaction

Mid-infrared laser filaments in the atmosphere

PubMed Central

Mitrofanov, A. V.; Voronin, A. A.; Sidorov-Biryukov, D. A.; Pugžlys, A.; Stepanov, E. A.; Andriukaitis, G.; Flöry, T.; Ališauskas, S.; Fedotov, A. B.; Baltuška, A.; Zheltikov, A. M.

2015-01-01

Filamentation of ultrashort laser pulses in the atmosphere offers unique opportunities for long-range transmission of high-power laser radiation and standoff detection. With the critical power of self-focusing scaling as the laser wavelength squared, the quest for longer-wavelength drivers, which would radically increase the peak power and, hence, the laser energy in a single filament, has been ongoing over two decades, during which time the available laser sources limited filamentation experiments in the atmosphere to the near-infrared and visible ranges. Here, we demonstrate filamentation of ultrashort mid-infrared pulses in the atmosphere for the first time. We show that, with the spectrum of a femtosecond laser driver centered at 3.9 μm, right at the edge of the atmospheric transmission window, radiation energies above 20 mJ and peak powers in excess of 200 GW can be transmitted through the atmosphere in a single filament. Our studies reveal unique properties of mid-infrared filaments, where the generation of powerful mid-infrared supercontinuum is accompanied by unusual scenarios of optical harmonic generation, giving rise to remarkably broad radiation spectra, stretching from the visible to the mid-infrared. PMID:25687621

The Connection Between Solar Coronal Cavities and Solar Filaments

NASA Astrophysics Data System (ADS)

Zawadzki, B.; Karna, N.; Prchlik, J.; Reeves, K.; Kempton, D.; Angryk, R.

2017-12-01

Filaments are structures in the solar corona made up of relatively cool, dense, partially ionized plasma. Coronal cavities, circular or elliptical regions of low plasma density, are observed above prominences on the solar limb when viewed in EUV and white light coronal images. Since most filament/cavity eruptions lead to a coronal mass ejection (CME), determining the likelihood of an eruption event will improve our ability to predict space weather. We examine SDO/AIA cavity metadata and HEK filament metadata to determine which cavities are associated with which filaments from 2012 to 2015. Our study involved 140 cavities and 368 filaments that appeared poleward of +-30 degrees. We categorized the cavities and filaments based on the stability of the structures, defined by whether or not the cavity and filament exist long enough to track fully across the solar disk. Using these categories we perform a statistical study on various filament qualities within the metadata. Our findings indicate that filaments with cavities are observed more often at high latitude in compared to filaments without cavities. Moreover, our study indicates that a statistically significant difference exists between the filament length and tilt distributions for certain categories. This work supported by the NSF-REU solar physics program at SAO, grant number AGS-1560313, and the NSF-DIBBS project, grant number ACI-1443061.

Filament winding technique, experiment and simulation analysis on tubular structure

NASA Astrophysics Data System (ADS)

Quanjin, Ma; Rejab, M. R. M.; Kaige, Jiang; Idris, M. S.; Harith, M. N.

2018-04-01

Filament winding process has emerged as one of the potential composite fabrication processes with lower costs. Filament wound products involve classic axisymmetric parts (pipes, rings, driveshafts, high-pressure vessels and storage tanks), non-axisymmetric parts (prismatic nonround sections and pipe fittings). Based on the 3-axis filament winding machine has been designed with the inexpensive control system, it is completely necessary to make a relative comparison between experiment and simulation on tubular structure. In this technical paper, the aim of this paper is to perform a dry winding experiment using the 3-axis filament winding machine and simulate winding process on the tubular structure using CADWIND software with 30°, 45°, 60° winding angle. The main result indicates that the 3-axis filament winding machine can produce tubular structure with high winding pattern performance with different winding angle. This developed 3-axis winding machine still has weakness compared to CAWIND software simulation results with high axes winding machine about winding pattern, turnaround impact, process error, thickness, friction impact etc. In conclusion, the 3-axis filament winding machine improvements and recommendations come up with its comparison results, which can intuitively understand its limitations and characteristics.

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

Transition from linear- to nonlinear-focusing regime in filamentation

PubMed Central

Lim, Khan; Durand, Magali; Baudelet, Matthieu; Richardson, Martin

2014-01-01

Laser filamentation in gases is often carried out in the laboratory with focusing optics to better stabilize the filament, whereas real-world applications of filaments frequently involve collimated or near-collimated beams. It is well documented that geometrical focusing can alter the properties of laser filaments and, consequently, a transition between a collimated and a strongly focused filament is expected. Nevertheless, this transition point has not been identified. Here, we propose an analytical method to determine the transition, and show that it corresponds to an actual shift in the balance of physical mechanisms governing filamentation. In high-NA conditions, filamentation is primarily governed by geometrical focusing and plasma effects, while the Kerr nonlinearity plays a more significant role as NA decreases. We find the transition between the two regimes to be relatively insensitive to the intrinsic laser parameters, and our analysis agrees well with a wide range of parameters found in published literature. PMID:25434678

Thioredoxin is required for filamentous phage assembly.

PubMed Central

Russel, M; Model, P

1985-01-01

Sequence comparisons show that the fip gene product of Escherichia coli, which is required for filamentous phage assembly, is thioredoxin. Thioredoxin serves as a cofactor for reductive processes in many cell types and is a constituent of phage T7 DNA polymerase. The fip-1 mutation makes filamentous phage and T7 growth temperature sensitive in cells that carry it. The lesion lies within a highly conserved thioredoxin active site. Thioredoxin reductase (NADPH), as well as thioredoxin, is required for efficient filamentous phage production. Mutant phages defective in phage gene I are particularly sensitive to perturbations in the fip-thioredoxin system. A speculative model is presented in which thioredoxin reductase, thioredoxin, and the gene I protein interact to drive an engine for filamentous phage assembly. Images PMID:3881756

Filamentation effect in a gas attenuator for high-repetition-rate X-ray FELs.

PubMed

Feng, Yiping; Krzywinski, Jacek; Schafer, Donald W; Ortiz, Eliazar; Rowen, Michael; Raubenheimer, Tor O

2016-01-01

A sustained filamentation or density depression phenomenon in an argon gas attenuator servicing a high-repetition femtosecond X-ray free-electron laser has been studied using a finite-difference method applied to the thermal diffusion equation for an ideal gas. A steady-state solution was obtained by assuming continuous-wave input of an equivalent time-averaged beam power and that the pressure of the entire gas volume has reached equilibrium. Both radial and axial temperature/density gradients were found and describable as filamentation or density depression previously reported for a femtosecond optical laser of similar attributes. The effect exhibits complex dependence on the input power, the desired attenuation, and the geometries of the beam and the attenuator. Time-dependent simulations were carried out to further elucidate the evolution of the temperature/density gradients in between pulses, from which the actual attenuation received by any given pulse can be properly calculated.

The Cape Ghir filament system in August 2009 (NW Africa)

NASA Astrophysics Data System (ADS)

Sangrà, Pablo; Troupin, Charles; Barreiro-González, Beatriz; Desmond Barton, Eric; Orbi, Abdellatif; Arístegui, Javier

2015-06-01

In the framework of the Canaries-Iberian marine ecosystem Exchanges (CAIBEX) experiment, an interdisciplinary high-resolution survey was conducted in the NW African region of Cape Ghir (30°38'N) during August 2009. The anatomy of a major filament is investigated on scales down to the submesoscale using in situ and remotely sensed data. The filament may be viewed as a system composed of three intimately connected structures: a small, shallow, and cold filament embedded within a larger, deeper, and cool filament and an intrathermocline anticyclonic eddy (ITE). The cold filament, which stretches 110 km offshore, is a shallow feature 60 m deep and 25 km wide, identified by minimal surface temperatures and rich in chlorophyll a. This structure comprises two asymmetrical submesoscale (˜18 km) fronts with jets flowing in opposite directions. The cold filament is embedded near the equatorward boundary of a much broader region of approximately 120 km width and 150 m depth that forms the cool filament and stretches at least 200 km offshore. This cool region, partly resulting from the influence of cold filament, is limited by two asymmetrical mesoscale (˜50 km) frontal boundaries. At the ITE, located north of the cold filament, we observe evidence of downwelling as indicated by a relatively high concentration of particles extending from the surface to more than 200 m depth. We hypothesize that this ITE may act as a sink of carbon and thus the filament system may serve dual roles of offshore carbon export and carbon sink.

In situ ecophysiology of Aigarchaeota from an oxic, hot-spring filamentous 'streamer' community

NASA Astrophysics Data System (ADS)

Beam, J.; Jay, Z.; Tringe, S. G.; Glavina del Rio, T.; Rusch, D.; Schmid, M.; Wagner, M.; Inskeep, W.

2014-12-01

The candidate phylum Aigarchaeota contains thermophilic archaea from terrestrial, subsurface, and marine geothermal ecosystems. The phylogeny and metabolic potential of Aigarchaeota has been deduced from several recent single-cell amplified genomes; however, an accurate description of their metabolism, potential ecological interactions, and role in biogeochemical cycling is lacking. Here we report possible ecological interactions and the in situ metabolism of an uncultivated lineage of Aigarchaeota from an oxic, terrestrial hot-spring filamentous 'streamer' community (Octopus Spring, pH = 8; T = 78 - 84 °C, Yellowstone National Park, Wyoming, USA). Fluorescence in situ hybridization (FISH) was combined with detailed genomic and transcriptomic reconstruction to elucidate the ecophysiological role of Aigarchaeota in these streamer communities. This novel population of Aigarchaeota are filamentous (~500 nm diameter by ~10-30 μm length), which is consistent with the morphology predicted by the presence and transcription of a single actin-encoding gene. Aigarchaeota filaments are intricately associated with other community members, which include both thermophilic bacteria and archaea. Metabolic reconstruction suggests that this aigarchaeon is an aerobic, chemoorganotroph. A single heme copper oxidase complex was identified in de novo genome assemblies, and was highly transcribed in environmental samples. Potential electron donors include acetate, fatty acids, sugars, peptides, and aromatic compounds. Transcripts related to genes specific to each of these potential electron donors were identified, indicating that this population of Aigarchaeota likely utilizes a broad range of reduced carbon substrates. Potential electron donors for this population may include extracellular polymeric substances produced by other microorganisms in close proximity. Flagellum genes were also highly transcribed, which suggests a potential mechanism for motility and/or cell-cell attachment

Heat transfer mechanism with thin filaments including ceramic high temperature heat exchanger

DOEpatents

Im, Kwan H.; Ahluwalia, Rajesh K.

1994-01-01

A radiative heat transfer mechanism in a furnace having burners through which pulverized coal and air are burned producing combustion gases and contaminants. A plurality of elongated conduits are positioned inside the furnace proximate to the burners generally parallel to the flow of combustion gases in the furnace. A plurality of thin filaments are inside each of the elongated hollow conduits, the filaments having diameters in the range of from about 1 micrometer to about 1,000 micrometers and having an infrared radiation cross-section sufficient to cause the filaments to heat upon exposure to infrared radiation. Blower mechanism is associated with the elongated conduits for limiting the amount of soot and ash which deposit on the conduits to preserve the radiative and convective transfer of heat energy from the combustion gases to the conduits.

Attachment and prejudice.

PubMed

Carnelley, Katherine B; Boag, Elle M

2018-04-16

There is a paucity of research that examines prejudice from an attachment theory perspective. Herein we make theoretical links between attachment patterns and levels of prejudice. Perceptions of outgroup threat, which activate the attachment system, are thought to lead to fear and prejudice for those high in attachment anxiety, and to distancing and prejudice for those high in attachment avoidance. We review the literature that examines the associations between attachment patterns and prejudice; evidence from attachment priming studies suggests a causal role of attachment security in reducing prejudice. We identify several mediators of these links: empathy, negative emotions, trust, social dominance orientation, romanticism, and contact quality. Future research should manipulate potential mediators and use psychophysiological assessments of threat. Copyright © 2018 Elsevier Ltd. All rights reserved.

Filamentation effect in a gas attenuator for high-repetition-rate X-ray FELs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Feng, Yiping; Krzywinski, Jacek; Schafer, Donald W.

A sustained filamentation or density depression phenomenon in an argon gas attenuator servicing a high-repetition femtosecond X-ray free-electron laser has been studied using a finite-difference method applied to the thermal diffusion equation for an ideal gas. A steady-state solution was obtained by assuming continuous-wave input of an equivalent time-averaged beam power and that the pressure of the entire gas volume has reached equilibrium. Both radial and axial temperature/density gradients were found and describable as filamentation or density depression previously reported for a femtosecond optical laser of similar attributes. The effect exhibits complex dependence on the input power, the desired attenuation,more » and the geometries of the beam and the attenuator. Time-dependent simulations were carried out to further elucidate the evolution of the temperature/density gradients in between pulses, from which the actual attenuation received by any given pulse can be properly calculated.« less

Involvement of the Vacuoles of the Endodermis in the Early Process of Shoot Gravitropism in Arabidopsis

PubMed Central

Morita, Miyo Terao; Kato, Takehide; Nagafusa, Kiyoshi; Saito, Chieko; Ueda, Takashi; Nakano, Akihiko; Tasaka, Masao

2002-01-01

The endodermal cells of the shoot are thought to be the gravity-sensing cells in Arabidopsis. The amyloplasts in the endodermis that sediment in the direction of gravity may act as statoliths. Endodermis-specific expression of SGR2 and ZIG using the SCR promoter could complement the abnormal shoot gravitropism of the sgr2 and zig mutants, respectively. The abnormalities in amyloplast sedimentation observed in both mutants recovered simultaneously. These results indicate that both genes in the endodermal cell layer are crucial for shoot gravitropism. ZIG encodes AtVTI11, which is a SNARE involved in vesicle transport to the vacuole. The fusion protein of SGR2 and green fluorescent protein localized to the vacuole and small organelles. These observations indicate that ZIG and SGR2 are involved in the formation and function of the vacuole, a notion supported by the results of subcellular analysis of the sgr2 and zig mutants with electron microscopy. These results strongly suggest that the vacuole participates in the early events of gravitropism and that SGR2 and ZIG functions are involved. PMID:11826298

Model-Assisted Analysis of Sugar Metabolism throughout Tomato Fruit Development Reveals Enzyme and Carrier Properties in Relation to Vacuole Expansion[W

PubMed Central

Beauvoit, Bertrand P.; Colombié, Sophie; Monier, Antoine; Andrieu, Marie-Hélène; Biais, Benoit; Bénard, Camille; Chéniclet, Catherine; Dieuaide-Noubhani, Martine; Nazaret, Christine; Mazat, Jean-Pierre; Gibon, Yves

2014-01-01

A kinetic model combining enzyme activity measurements and subcellular compartmentation was parameterized to fit the sucrose, hexose, and glucose-6-P contents of pericarp throughout tomato (Solanum lycopersicum) fruit development. The model was further validated using independent data obtained from domesticated and wild tomato species and on transgenic lines. A hierarchical clustering analysis of the calculated fluxes and enzyme capacities together revealed stage-dependent features. Cell division was characterized by a high sucrolytic activity of the vacuole, whereas sucrose cleavage during expansion was sustained by both sucrose synthase and neutral invertase, associated with minimal futile cycling. Most importantly, a tight correlation between flux rate and enzyme capacity was found for fructokinase and PPi-dependent phosphofructokinase during cell division and for sucrose synthase, UDP-glucopyrophosphorylase, and phosphoglucomutase during expansion, thus suggesting an adaptation of enzyme abundance to metabolic needs. In contrast, for most enzymes, flux rates varied irrespectively of enzyme capacities, and most enzymes functioned at <5% of their maximal catalytic capacity. One of the major findings with the model was the high accumulation of soluble sugars within the vacuole together with organic acids, thus enabling the osmotic-driven vacuole expansion that was found during cell division. PMID:25139005

Control of multiple filamentation in air

NASA Astrophysics Data System (ADS)

Fibich, Gadi; Eisenmann, Shmuel; Ilan, Boaz; Zigler, Arie

2004-08-01

In this Letter we provide what is believed to be the first experimental evidence of suppression of the number of filaments for high-intensity laser pulses propagating in air by beam astigmatism. We also show that the number, pattern, and spatial stability of the filaments can be controlled by varying the angle that a focusing lens makes with the axial direction of propagation. This new methodology can be useful for applications involving atmospheric propagation, such as remote sensing.

Heat transfer mechanism with thin filaments including ceramic high temperature heat exchanger

DOEpatents

Im, K.H.; Ahluwalia, R.K.

1994-10-18

A radiative heat transfer mechanism in a furnace is described having burners through which pulverized coal and air are burned producing combustion gases and contaminants. A plurality of elongated conduits are positioned inside the furnace proximate to the burners generally parallel to the flow of combustion gases in the furnace. A plurality of thin filaments are inside each of the elongated hollow conduits, the filaments having diameters in the range of from about 1 micrometer to about 1,000 micrometers and having an infrared radiation cross-section sufficient to cause the filaments to heat upon exposure to infrared radiation. Blower mechanism is associated with the elongated conduits for limiting the amount of soot and ash which deposit on the conduits to preserve the radiative and convective transfer of heat energy from the combustion gases to the conduits. 7 figs.

CYTOPLASMIC FILAMENTS OF AMOEBA PROTEUS

PubMed Central

Pollard, Thomas D.; Ito, Susumu

1970-01-01

The role of filaments in consistency changes and movement in a motile cytoplasmic extract of Amoeba proteus was investigated by correlating light and electron microscopic observations with viscosity measurements. The extract is prepared by the method of Thompson and Wolpert (1963). At 0°C, this extract is nonmotile and similar in structure to ameba cytoplasm, consisting of groundplasm, vesicles, mitochondria, and a few 160 A filaments. The extract undergoes striking ATP-stimulated streaming when warmed to 22°C. Two phases of movement are distinguished. During the first phase, the apparent viscosity usually increases and numerous 50–70 A filaments appear in samples of the extract prepared for electron microscopy, suggesting that the increase in viscosity in caused, at least in part, by the formation of these thin filaments. During this initial phase of ATP-stimulated movement, these thin filaments are not detectable by phase-contrast or polarization microscopy, but later, in the second phase of movement, 70 A filaments aggregate to form birefringent microscopic fibrils. A preparation of pure groundplasm with no 160 A filaments or membranous organelles exhibits little or no ATP-stimulated movement, but 50–70 A filaments form and aggregate into birefringent fibrils. This observation and the structural relationship of the 70 A and the 160 A filaments in the motile extract suggest that both types of filaments may be required for movement. These two types of filaments, 50–70 A and 160 A, are also present in the cytoplasm of intact amebas. Fixed cells could not be used to study the distribution of these filaments during natural ameboid movement because of difficulties in preserving the normal structure of the ameba during preparation for electron microscopy. PMID:4915451

[Effect of acetylcholine and acetylcholinesterase on the activity of contractile vacuole of Amoeba proteus].

PubMed

Bagrov, Ia Iu; Manusova, N B

2011-01-01

Acetylcholine (ACh, 1 microM) stimulates activity of the contractile vacuole of proteus. The effect of ACh is not mimicked by its analogs which are not hydrolyzed by acetylcholinesterase (AChE), i. e., carbacholine and 5-methylfurmethide. The effect of ACh is not sensitive to the blocking action of M-cholinolytics, atropine and mytolone, but is suppressed by N-cholinolytic, tubocurarine. The inhibitors of AChE, eserine (0.01 microM) and armine (0.1 microM), suppress the effect of ACh on amoeba contractile vacuole. ACh does not affect activation of contractile vacuole induced by arginine-vasopressin (1 microM), but it blocks such effect of opiate receptors agonist, dynorphin A1-13 (0.01 microM). This effect of ACh is also suppressed by the inhibitors of AChE. These results suggest that, in the above-described effects of ACh, AChE acts not as an antagonist, but rather as a synergist.

Changes in vacuolation in the root apex cells of soybean seedlings in microgravity

NASA Technical Reports Server (NTRS)

Klymchuk, D. O.; Kordyum, E. L.; Vorobyova, T. V.; Chapman, D. K.; Brown, C. S.

2003-01-01

Changes in the vacuolation in root apex cells of soybean (Glycine max L. [Merr.]) seedlings grown in microgravity were investigated. Spaceflight and ground control seedlings were grown in the absence or presence of KMnO4 (to remove ethylene) for 6 days. After landing, in order to study of cell ultrastructure and subcellular free calcium ion distribution, seedling root apices were fixed in 2.5% (w/v) glutaraldehyde in 0.1 M cacodylate buffer and 2% (w/v) glutaraldehyde, 2.5% (w/v) formaldehyde, 2% (w/v) potassium antimonate K[Sb(OH)6] in 0.1 M K2HPO4 buffer with an osmolarity (calculated theoretically) of 0.45 and 1.26 osmol. The concentrations of ethylene in all spaceflight canisters were significantly higher than in the ground control canisters. Seedling growth was reduced in the spaceflight-exposed plants. Additionally, the spaceflight-exposed plants exhibited progressive vacuolation in the root apex cells, particularly in the columella cells, to a greater degree than the ground controls. Plasmolysis was observed in columella cells of spaceflight roots fixed in solutions with relatively high osmolarity (1.26 osmol). The appearance of plasmolysis permitted the evaluation of the water status of cells. The water potential of the spaceflight cells was higher than the surrounding fixative solution. A decrease in osmotic potential and/or an increase in turgor potential may have induced increases in cell water potential. However, the plasmolysed (i.e. non-turgid) cells implied that increases in water potential were accompanied with a decrease in osmotic potential. In such cells changes in vacuolation may have been involved to maintain turgor pressure or may have been a result of intensification of other vacuolar functions like digestion and storage. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

Femtosecond laser fabrication of sub-diffraction nanoripples on wet Al surface in multi-filamentation regime: High optical harmonics effects?

NASA Astrophysics Data System (ADS)

Ionin, A. A.; Kudryashov, S. I.; Makarov, S. V.; Rudenko, A. A.; Saltuganov, P. N.; Seleznev, L. V.; Sinitsyn, D. V.; Sunchugasheva, E. S.

2014-02-01

Relief ripples with sub-diffraction periods (≈λlas/3, λlas/4) were produced on a aluminum surface immersed in water and irradiated in a multi-filamentation regime by focused 744 nm femtosecond laser pulses with highly supercritical, multi-GW peak powers. For the VUV (8.5 eV) surface plasmon resonance on the wet aluminum surface, such small-scale surface nanogratings can be produced by high - second and third - optical harmonics, coming to the surface from the optical filaments in the water layer. Then, the sub-diffraction surface ripples may appear through interference of their transverse electric fields with the longitudinal electric fields of their counterparts, scattered on the surface roughness and appeared as the corresponding high-energy, high-wavenumber surface polaritons.

Cytoplasmic filaments of Amoeba proteus. I. The role of filaments in consistency changes and movement.

PubMed

Pollard, T D; Ito, S

1970-08-01

The role of filaments in consistency changes and movement in a motile cytoplasmic extract of Amoeba proteus was investigated by correlating light and electron microscopic observations with viscosity measurements. The extract is prepared by the method of Thompson and Wolpert (1963). At 0 degrees C, this extract is nonmotile and similar in structure to ameba cytoplasm, consisting of groundplasm, vesicles, mitochondria, and a few 160 A filaments. The extract undergoes striking ATP-stimulated streaming when warmed to 22 degrees C. Two phases of movement are distinguished. During the first phase, the apparent viscosity usually increases and numerous 50-70 A filaments appear in samples of the extract prepared for electron microscopy, suggesting that the increase in viscosity in caused, at least in part, by the formation of these thin filaments. During this initial phase of ATP-stimulated movement, these thin filaments are not detectable by phase-contrast or polarization microscopy, but later, in the second phase of movement, 70 A filaments aggregate to form birefringent microscopic fibrils. A preparation of pure groundplasm with no 160 A filaments or membranous organelles exhibits little or no ATP-stimulated movement, but 50-70 A filaments form and aggregate into birefringent fibrils. This observation and the structural relationship of the 70 A and the 160 A filaments in the motile extract suggest that both types of filaments may be required for movement. These two types of filaments, 50-70 A and 160 A, are also present in the cytoplasm of intact amebas. Fixed cells could not be used to study the distribution of these filaments during natural ameboid movement because of difficulties in preserving the normal structure of the ameba during preparation for electron microscopy.

Dynamics and Breakup of a Contracting Viscous Filament

NASA Astrophysics Data System (ADS)

Wilkes, Edward; Notz, Patrick; Ambravaneswaran, Bala; Basaran, Osman

1999-11-01

Free viscous filaments are formed during the breakup of liquid drops and jets. Such filaments are typically precursors of satellite droplets that are often undesirable in applications such as ink-jet printing. In this paper, the contraction of an axisymmetric liquid filament due to action of surface tension is studied theoretically. The analysis is based on solving (a) the full Navier-Stokes system in two-dimensions (2-d) and (b) a one-dimensional (1-d) approximation of the exact equations based on slender-jet theory. The rigorous, 2-d calculations are carried out with finite element algorithms using either algebraic or elliptic mesh generation. As the filament contracts, bulbous regions form at its two ends. When the initial aspect ratio a/b and/or the Reynolds number Re are sufficiently low, the ends coalesce into an oscillating free drop. Filament breakup occurs when a/b and/or Re are sufficiently high. The 2-d algorithms reveal for the first time that liquid filaments of finite viscosity can overturn prior to interface rupture. The power of elliptic mesh generation over algebraic methods in analyzing such situations is highlighted.

The vacuole model: new terms in the second order deflection of light

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bhattacharya, Amrita; Nandi, Kamal K.; Garipova, Guzel M.

2011-02-01

The present paper is an extension of a recent work (Bhattacharya et al. 2010) to the Einstein-Strauss vacuole model with a cosmological constant, where we work out the light deflection by considering perturbations up to order M{sup 3} and confirm the light bending obtained previously in their vacuole model by Ishak et al. (2008). We also obtain another local coupling term −5πM{sup 2}Λ/8 related to Λ, in addition to the one obtained by Sereno (2008, 2009). We argue that the vacuole method for light deflection is exclusively suited to cases where the cosmological constant Λ disappears from the path equation.more » However, the original Rindler-Ishak method (2007) still applies even if a certain parameter γ of Weyl gravity does not disappear. Here, using an alternative prescription, we obtain the known term −γR/2, as well as another new local term 3πγM/2 between M and γ. Physical implications are compared, where we argue that the repulsive term −γR/2 can be masked by the Schwarzschild term 2M/R in the halo regime supporting attractive property of the dark matter.« less

Solid friction between soft filaments.

PubMed

Ward, Andrew; Hilitski, Feodor; Schwenger, Walter; Welch, David; Lau, A W C; Vitelli, Vincenzo; Mahadevan, L; Dogic, Zvonimir

2015-06-01

Any macroscopic deformation of a filamentous bundle is necessarily accompanied by local sliding and/or stretching of the constituent filaments. Yet the nature of the sliding friction between two aligned filaments interacting through multiple contacts remains largely unexplored. Here, by directly measuring the sliding forces between two bundled F-actin filaments, we show that these frictional forces are unexpectedly large, scale logarithmically with sliding velocity as in solid-like friction, and exhibit complex dependence on the filaments' overlap length. We also show that a reduction of the frictional force by orders of magnitude, associated with a transition from solid-like friction to Stokes's drag, can be induced by coating F-actin with polymeric brushes. Furthermore, we observe similar transitions in filamentous microtubules and bacterial flagella. Our findings demonstrate how altering a filament's elasticity, structure and interactions can be used to engineer interfilament friction and thus tune the properties of fibrous composite materials.

Flux Cancellation Leading to Solar Filament Eruptions

NASA Astrophysics Data System (ADS)

Popescu, R. M.; Panesar, N. K.; Sterling, A. C.; Moore, R. L.

2016-12-01

Solar filaments are strands of relatively cool, dense plasma magnetically suspended in the lower density hotter solar corona. They trace magnetic polarity inversion lines (PILs) in the photosphere below, and are supported against gravity at heights of up to 100 Mm above the chromosphere by the magnetic field in and around them. This field erupts when it is rendered unstable by either magnetic flux cancellation or emergence at or near the PIL. We have studied the evolution of photospheric magnetic flux leading to ten observed filament eruptions. Specifically, we look for gradual magnetic changes in the neighborhood of the PIL prior to and during eruption. We use Extreme Ultraviolet (EUV) images from the Atmospheric Imaging Assembly (AIA), and magnetograms from the Helioseismic and Magnetic Imager (HMI), both onboard the Solar Dynamics Observatory (SDO), to study filament eruptions and their photospheric magnetic fields. We examine whether flux cancellation or/and emergence leads to filament eruptions and find that continuous flux cancellation was present at the PIL for many hours prior to each eruption. We present two events in detail and find the following: (a) the pre-eruption filament-holding core field is highly sheared and appears in the shape of a sigmoid above the PIL; (b) at the start of the eruption the opposite arms of the sigmoid reconnect in the middle above the site of (tether-cutting) flux cancellation at the PIL; (c) the filaments first show a slow-rise, followed by a fast-rise as they erupt. We conclude that these two filament eruptions result from flux cancellation in the middle of the sheared field and are in agreement with the standard model for a CME/flare filament eruption from a closed bipolar magnetic field [flux cancellation (van Ballegooijen and Martens 1989 and Moore and Roumelrotis 1992) and runaway tether-cutting (Moore et. al 2001)].

Flux Cancellation Leading to CME Filament Eruptions

NASA Technical Reports Server (NTRS)

Popescu, Roxana M.; Panesar, Navdeep K.; Sterling, Alphonse C.; Moore, Ronald L.

2016-01-01

Solar filaments are strands of relatively cool, dense plasma magnetically suspended in the lower density hotter solar corona. They trace magnetic polarity inversion lines (PILs) in the photosphere below, and are supported against gravity at heights of up to approx.100 Mm above the chromosphere by the magnetic field in and around them. This field erupts when it is rendered unstable, often by magnetic flux cancellation or emergence at or near the PIL. We have studied the evolution of photospheric magnetic flux leading to ten observed filament eruptions. Specifically, we look for gradual magnetic changes in the neighborhood of the PIL prior to and during eruption. We use Extreme Ultraviolet (EUV) images from the Atmospheric Imaging Assembly (AIA), and magnetograms from the Helioseismic and Magnetic Imager (HMI), both on board the Solar Dynamics Observatory (SDO), to study filament eruptions and their photospheric magnetic fields. We examine whether flux cancellation or/and emergence leads to filament eruptions. We find that continuous flux cancellation was present at the PIL for many hours prior to each eruption. We present two CME-producing eruptions in detail and find the following: (a) the pre-eruption filament-holding core field is highly sheared and appears in the shape of a sigmoid above the PIL; (b) at the start of the eruption the opposite arms of the sigmoid reconnect in the middle above the site of (tether-cutting) flux cancellation at the PIL; (c) the filaments first show a slow-rise, followed by a fast-rise as they erupt. We conclude that these two filament eruptions result from flux cancellation in the middle of the sheared field, and thereafter evolve in agreement with the standard model for a CME/flare filament eruption from a closed bipolar magnetic field [flux cancellation (van Ballegooijen and Martens 1989 and Moore and Roumelrotis 1992) and runaway tether-cutting (Moore et. al 2001)].

Characterization of somatic embryo attached structures in Feijoa sellowiana Berg. (Myrtaceae).

PubMed

Correia, Sandra M; Canhoto, Jorge M

2010-06-01

The presence of an attached organ to somatic embryos of angiosperms connecting the embryo to the supporting tissue has been a subject of controversy. This study shows that 67% of the morphologically normal somatic embryos of Feijoa sellowiana possess this type of organ and that its formation was not affected by culture media composition. Histological and ultrastructural analysis indicated that the attached structures of somatic embryos displayed a great morphological diversity ranging from a few cells to massive and columnar structures. This contrast with the simple suspensors observed in zygotic embryos which were only formed by five cells. As well as the suspensor of zygotic embryos, somatic embryo attached structures undergo a process of degeneration in later stages of embryo development. Other characteristic shared by zygotic suspensors and somatic embryo attached structures was the presence of thick cell walls surrounding the cells. Elongated thin filaments were often associated with the structures attached to somatic embryos, whereas in other cases, tubular cells containing starch grains connected the embryo to the supporting tissue. These characteristics associated with the presence of plasmodesmata in the cells of the attached structures seem to indicate a role on embryo nutrition. However, cell proliferation in the attached structures resulting into new somatic embryos may also suggest a more complex relationship between the embryo and the structures connecting it to the supporting tissue.

Electron microscopic evidence for the myosin head lever arm mechanism in hydrated myosin filaments using the gas environmental chamber

DOE Office of Scientific and Technical Information (OSTI.GOV)

Minoda, Hiroki; CREST, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012; Okabe, Tatsuhiro

2011-02-25

Research highlights: {yields} We succeeded in recording structural changes of hydrated myosin cross-bridges. {yields} We succeeded in position-marking the cross-bridges with site-directed antibodies. {yields} We recorded cross-bridge movement at different regions in individual cross-bridge. {yields} The movement was smallest at the cross-bridge-subfragment two boundary. {yields} The results provide evidence for the cross-bridge lever arm mechanism. -- Abstract: Muscle contraction results from an attachment-detachment cycle between the myosin heads extending from myosin filaments and the sites on actin filaments. The myosin head first attaches to actin together with the products of ATP hydrolysis, performs a power stroke associated with release ofmore » hydrolysis products, and detaches from actin upon binding with new ATP. The detached myosin head then hydrolyses ATP, and performs a recovery stroke to restore its initial position. The strokes have been suggested to result from rotation of the lever arm domain around the converter domain, while the catalytic domain remains rigid. To ascertain the validity of the lever arm hypothesis in muscle, we recorded ATP-induced movement at different regions within individual myosin heads in hydrated myosin filaments, using the gas environmental chamber attached to the electron microscope. The myosin head were position-marked with gold particles using three different site-directed antibodies. The amplitude of ATP-induced movement at the actin binding site in the catalytic domain was similar to that at the boundary between the catalytic and converter domains, but was definitely larger than that at the regulatory light chain in the lever arm domain. These results are consistent with the myosin head lever arm mechanism in muscle contraction if some assumptions are made.« less

Evaluation of filamentous green algae as feedstocks for biofuel production.

PubMed

Zhang, Wei; Zhao, Yonggang; Cui, Binjie; Wang, Hui; Liu, Tianzhong

2016-11-01

Compared with unicellular microalgae, filamentous algae have high resistance to grazer-predation and low-cost recovery in large-scale production. Green algae, as the most diverse group of algae, included numerous filamentous genera and species. In this study, records of filamentous genera and species in green algae were firstly censused and classified. Then, seven filamentous strains subordinated in different genera were cultivated in bubbled-column to investigate their growth rate and energy molecular (lipid and starch) capacity. Four strains including Stigeoclonium sp., Oedogonium nodulosum, Hormidium sp. and Zygnema extenue were screened out due to their robust growth. And they all could accumulate triacylglycerols and starch in their biomass, but with different capacity. After nitrogen starvation, Hormidium sp. and Oedogonium nodulosum respectively exhibited high capacity of lipid (45.38% in dry weight) and starch (46.19% in dry weight) accumulation, which could be of high potential as feedstocks for biodiesel and bioethanol production. Copyright © 2016 Elsevier Ltd. All rights reserved.

Patterns of Very Insecure Attachment Found in Samples of High-Risk Infants and Toddlers.

ERIC Educational Resources Information Center

Spieker, Susan J.

1986-01-01

The study examined the classification scheme of the Ainsworth Strange Situation measure of attachment security with 44 insecurely attached 13-month olds from a high-social-risk sample. Results supported the existence of an additional category of infants whose attachment behavior was "avoidant/ambivalent" and whose families were…

Cellular vacuolation and mitochondrial-associated factors induced by Clostridium perfringens epsilon toxin detected using acoustic flow cytometry.

PubMed

Ferrarezi, Marina C; Curci, Vera C L M; Cardoso, Tereza C

2013-12-01

Epsilon toxin (ETX) produced by Clostridium perfringens types B and D is a potent toxin that is responsible for fatal enterotoxaemia. In vitro, ETX, which is considered as a pore-forming toxin, forms a heptamer in Madin-Darby canine kidney (MDCK) cell membranes, which is considered to be a pre-pore stage. After binding of the ETX, vacuoles inside cell cytoplasm are produced. ETX causes decreased levels of essential coenzymes required for host cell energy. Here, we optimized and applied acoustic flow cytometry analysis in order to gain further insight into ETX-pathogenesis. Using acoustic flow cytometer analysis, which considered highly sensitive, ETX-exposed MDCK cells revealed mitochondrial membrane decreases followed by 25.48% and 45.45% of the exposed cells expressing the Bax and BCL-2 proteins at a pre-pore stage, respectively. These results together with high cytotoxicity and visualization of cell vacuoles, demonstrates that acoustic flow cytometry analysis potentially represents an effective tool to study ETX pathogenesis. Copyright © 2013. Published by Elsevier Ltd.

Design of the klystron filament power supply control system for EAST LHCD

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Zege; Wang, Mao; Hu, Huaichuan

A filament is a critical component of the klystron used to heat the cathode. There are totally 44 klystrons in experimental advanced superconducting tokamak (EAST) lower hybrid current drive (LHCD) systems. All klystron filaments are powered by AC power suppliers through isolated transformers. In order to achieve better klystron preheat, a klystron filament power supply control system is designed to obtain the automatic control of all filament power suppliers. Klystron filament current is measured by PLC and the interlock between filament current and klystron high voltage system is also implemented. This design has already been deployed in two LHCD systemsmore » and proves feasible completely.« less

Dry soldering with hot filament produced atomic hydrogen

DOEpatents

Panitz, Janda K. G.; Jellison, James L.; Staley, David J.

1995-01-01

A system for chemically transforming metal surface oxides to metal that is especially, but not exclusively, suitable for preparing metal surfaces for dry soldering and solder reflow processes. The system employs one or more hot, refractory metal filaments, grids or surfaces to thermally dissociate molecular species in a low pressure of working gas such as a hydrogen-containing gas to produce reactive species in a reactive plasma that can chemically reduce metal oxides and form volatile compounds that are removed in the working gas flow. Dry soldering and solder reflow processes are especially applicable to the manufacture of printed circuit boards, semiconductor chip lead attachment and packaging multichip modules. The system can be retrofitted onto existing metal treatment ovens, furnaces, welding systems and wave soldering system designs.

Nonequilibrium transport in superconducting filaments

NASA Technical Reports Server (NTRS)

Arutyunov, K. YU.; Danilova, N. P.; Nikolaeva, A. A.

1995-01-01

The step-like current-voltage characteristics of highly homogeneous single-crystalline tin and indium thin filaments has been measured. The length of the samples L approximately 1 cm was much greater than the nonequilibrium quasiparticle relaxation length Lambda. It was found that the activation of a successive i-th voltage step occurs at current significantly greater than the one derived with the assumption that the phase slip centers are weakly interacting on a scale L much greater than Lambda. The observation of 'subharmonic' fine structure on the voltage-current characteristics of tin filaments confirms the hypothesis of the long-range phase slip centers interaction.

Filamentous Morphology as a Means for Thermophilic Bacteria to Survive Steep Physical and Chemical Gradients in Yellowstone Hot Springs

NASA Astrophysics Data System (ADS)

Dong, Y.; Srivastava, V.; Bulone, V.; Keating, K. M.; Khetani, R. S.; Fields, C. J.; Inskeep, W.; Sanford, R. A.; Yau, P. M.; Imai, B. S.; Hernandez, A. G.; Wright, C.; Band, M.; Cann, I. K.; Ahrén, D.; Fouke, K. W.; Sivaguru, M.; Fried, G.; Fouke, B. W.

2017-12-01

The filamentous heat-loving bacterium Sulfurihydrogenibium yellowstonense makes up more than 90% of the microbial community that inhabits turbulent, dysoxic hot spring outflow channels (66-71°C, 6.2-6.5 pH, 0.5-0.75 m/s flow rate) at Mammoth Hot Spring in Yellowstone National Park. These environments contain abundantly available inorganic substrates (e.g., CO2, sulfide and thiosulfate) and are associated with extensive CaCO3 (travertine) precipitation driven in part by CO2 off-gassing. Evidence from integrated Meta-Omics analyses of DNA, RNA, and proteins (metagenomics, metatranscriptomics and metaproteomics) extracted from these S. yellowstonense-dominated communities have detected 1499 non-rRNA open reading frames (ORFs), their transcripts and cognate proteins. During chemoautotrophy and CO2 carbon fixation, chaperons facilitate enzymatic stability and functionalities under elevated temperature. High abundance transcripts and proteins for Type IV pili and exopolysaccharides (EPS) are consistent with S. yellowstonense forming strong (up to 0.5 m) intertwined microbial filaments (fettuccini streamers) composed of linked individual cells that withstand hydrodynamic shear forces and extremely rapid travertine mineralization. Their primary energy source is the oxidation of reduced sulfur (e.g., sulphide, sulfur or thiosulfate) and the simultaneous uptake of extremely low concentrations of dissolved O2 facilitated by bd-type cytochromes. Field observations indicate that the fettuccini microbial filaments build up ridged travertine platforms on the bottom of the springs, parallel to the water flow, where living filaments attach almost exclusively to the top of each ridge. This maximizes their access to miniscule amounts of dissolved oxygen, while optimizing their ability to rapidly form down-flow branched filaments and thus survive in these stressful environments that few other microbes can inhabit.

The bipolar filaments formed by Herpes simplex virus type 1 SSB/recombination protein (ICP8) suggest a mechanism for DNA annealing

PubMed Central

Makhov, Alexander M.; Sen, Anindito; Yu, Xiong; Simon, Martha N.; Griffith, Jack D.; Egelman, Edward H.

2009-01-01

Herpes simplex virus type 1 encodes a multifunctional protein, ICP8, which serves both as a single strand binding protein and recombinase, catalyzing reactions involved in replication and recombination of the viral genome. In the presence of divalent ions and at low temperature, previous electron microscopic (EM) studies showed that ICP8 will form long left-handed helical filaments. Here EM image reconstruction reveals that the filaments are bipolar, with an asymmetric unit containing two subunits of ICP8 that constitute a symmetrical dimer. This organization of the filament has been confirmed using Scanning Transmission Electron Microscopy. The pitch of the filaments is ~ 250 Å, with ~ 6.2 dimers per turn. Docking of a crystal structure of ICP8 into the reconstructed filament shows that the C-terminal domain of ICP8, attached to the body of the subunit by a flexible linker containing ~ 10 residues, is packed into a pocket in the body of a neighboring subunit in the crystal in a similar manner as in the filament. However, the interactions between the large N-terminal domains are quite different in the filament from that observed in the crystal. A previously proposed model for ICP8 binding single-stranded DNA, based upon the crystal structure, leads to a model for a continuous strand of ssDNA near the filament axis. The bipolar nature of the ICP8 filaments means that a second strand of ssDNA would be running through this filament in the opposite orientation, and this provides a potential mechanism for how ICP8 anneals complementary single stranded DNA into double-stranded DNA, where each strand runs in opposite directions. PMID:19138689

The bipolar filaments formed by herpes simplex virus type 1 SSB/recombination protein (ICP8) suggest a mechanism for DNA annealing.

PubMed

Makhov, Alexander M; Sen, Anindito; Yu, Xiong; Simon, Martha N; Griffith, Jack D; Egelman, Edward H

2009-02-20

Herpes simplex virus type 1 encodes a multifunctional protein, ICP8, which serves both as a single-strand binding protein and as a recombinase, catalyzing reactions involved in replication and recombination of the viral genome. In the presence of divalent ions and at low temperature, previous electron microscopic studies showed that ICP8 will form long left-handed helical filaments. Here, electron microscopic image reconstruction reveals that the filaments are bipolar, with an asymmetric unit containing two subunits of ICP8 that constitute a symmetrical dimer. This organization of the filament has been confirmed using scanning transmission electron microscopy. The pitch of the filaments is approximately 250 A, with approximately 6.2 dimers per turn. Docking of a crystal structure of ICP8 into the reconstructed filament shows that the C-terminal domain of ICP8, attached to the body of the subunit by a flexible linker containing approximately 10 residues, is packed into a pocket in the body of a neighboring subunit in the crystal in a similar manner as in the filament. However, the interactions between the large N-terminal domains are quite different in the filament from that observed in the crystal. A previously proposed model for ICP8 binding single-stranded DNA (ssDNA), based upon the crystal structure, leads to a model for a continuous strand of ssDNA near the filament axis. The bipolar nature of the ICP8 filaments means that a second strand of ssDNA would be running through this filament in the opposite orientation, and this provides a potential mechanism for how ICP8 anneals complementary ssDNA into double-stranded DNA, where each strand runs in opposite directions.

Adjustment of Host Cells for Accommodation of Symbiotic Bacteria: Vacuole Defunctionalization, HOPS Suppression, and TIP1g Retargeting in Medicago[C][W][OPEN

PubMed Central

Gavrin, Aleksandr; Kaiser, Brent N.; Geiger, Dietmar; Tyerman, Stephen D.; Wen, Zhengyu; Bisseling, Ton; Fedorova, Elena E.

2014-01-01

In legume–rhizobia symbioses, the bacteria in infected cells are enclosed in a plant membrane, forming organelle-like compartments called symbiosomes. Symbiosomes remain as individual units and avoid fusion with lytic vacuoles of host cells. We observed changes in the vacuole volume of infected cells and thus hypothesized that microsymbionts may cause modifications in vacuole formation or function. To examine this, we quantified the volumes and surface areas of plant cells, vacuoles, and symbiosomes in root nodules of Medicago truncatula and analyzed the expression and localization of VPS11 and VPS39, members of the HOPS vacuole-tethering complex. During the maturation of symbiosomes to become N2-fixing organelles, a developmental switch occurs and changes in vacuole features are induced. For example, we found that expression of VPS11 and VPS39 in infected cells is suppressed and host cell vacuoles contract, permitting the expansion of symbiosomes. Trafficking of tonoplast-targeted proteins in infected symbiotic cells is also altered, as shown by retargeting of the aquaporin TIP1g from the tonoplast membrane to the symbiosome membrane. This retargeting appears to be essential for the maturation of symbiosomes. We propose that these alterations in the function of the vacuole are key events in the adaptation of the plant cell to host intracellular symbiotic bacteria. PMID:25217511

The Fab1/PIKfyve phosphoinositide phosphate kinase is not necessary to maintain the pH of lysosomes and of the yeast vacuole.

PubMed

Ho, Cheuk Y; Choy, Christopher H; Wattson, Christina A; Johnson, Danielle E; Botelho, Roberto J

2015-04-10

Lysosomes and the yeast vacuole are degradative and acidic organelles. Phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P2), a master architect of endolysosome and vacuole identity, is thought to be necessary for vacuolar acidification in yeast. There is also evidence that PtdIns(3,5)P2 may play a role in lysosomal acidification in higher eukaryotes. Nevertheless, these conclusions rely on qualitative assays of lysosome/vacuole pH. For example, quinacrine, an acidotropic fluorescent base, does not accumulate in the vacuoles of fab1Δ yeast. Fab1, along with its mammalian ortholog PIKfyve, is the lipid kinase responsible for synthesizing PtdIns(3,5)P2. In this study, we employed several assays that quantitatively assessed the lysosomal and vacuolar pH in PtdIns(3,5)P2-depleted cells. Using ratiometric imaging, we conclude that lysosomes retain a pH < 5 in PIKfyve-inhibited mammalian cells. In addition, quantitative fluorescence microscopy of vacuole-targeted pHluorin, a pH-sensitive GFP variant, indicates that fab1Δ vacuoles are as acidic as wild-type yeast. Importantly, we also employed fluorimetry of vacuoles loaded with cDCFDA, a pH-sensitive dye, to show that both wild-type and fab1Δ vacuoles have a pH < 5.0. In comparison, the vacuolar pH of the V-ATPase mutant vph1Δ or vph1Δ fab1Δ double mutant was 6.1. Although the steady-state vacuolar pH is not affected by PtdIns(3,5)P2 depletion, it may have a role in stabilizing the vacuolar pH during salt shock. Overall, we propose a model in which PtdIns(3,5)P2 does not govern the steady-state pH of vacuoles or lysosomes. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

The Fab1/PIKfyve Phosphoinositide Phosphate Kinase Is Not Necessary to Maintain the pH of Lysosomes and of the Yeast Vacuole*

PubMed Central

Ho, Cheuk Y.; Choy, Christopher H.; Wattson, Christina A.; Johnson, Danielle E.; Botelho, Roberto J.

2015-01-01

Lysosomes and the yeast vacuole are degradative and acidic organelles. Phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P2), a master architect of endolysosome and vacuole identity, is thought to be necessary for vacuolar acidification in yeast. There is also evidence that PtdIns(3,5)P2 may play a role in lysosomal acidification in higher eukaryotes. Nevertheless, these conclusions rely on qualitative assays of lysosome/vacuole pH. For example, quinacrine, an acidotropic fluorescent base, does not accumulate in the vacuoles of fab1Δ yeast. Fab1, along with its mammalian ortholog PIKfyve, is the lipid kinase responsible for synthesizing PtdIns(3,5)P2. In this study, we employed several assays that quantitatively assessed the lysosomal and vacuolar pH in PtdIns(3,5)P2-depleted cells. Using ratiometric imaging, we conclude that lysosomes retain a pH < 5 in PIKfyve-inhibited mammalian cells. In addition, quantitative fluorescence microscopy of vacuole-targeted pHluorin, a pH-sensitive GFP variant, indicates that fab1Δ vacuoles are as acidic as wild-type yeast. Importantly, we also employed fluorimetry of vacuoles loaded with cDCFDA, a pH-sensitive dye, to show that both wild-type and fab1Δ vacuoles have a pH < 5.0. In comparison, the vacuolar pH of the V-ATPase mutant vph1Δ or vph1Δ fab1Δ double mutant was 6.1. Although the steady-state vacuolar pH is not affected by PtdIns(3,5)P2 depletion, it may have a role in stabilizing the vacuolar pH during salt shock. Overall, we propose a model in which PtdIns(3,5)P2 does not govern the steady-state pH of vacuoles or lysosomes. PMID:25713145

Plethora of transitions during breakup of liquid filaments

DOE PAGES

Castrejón-Pita, José Rafael; Castrejón-Pita, Alfonso Arturo; Thete, Sumeet Suresh; ...

2015-03-30

Thinning and breakup of liquid filaments are central to dripping of leaky faucets, inkjet drop formation, and raindrop fragmentation. As the filament radius decreases, curvature and capillary pressure, both inversely proportional to radius, increase and fluid is expelled with increasing velocity from the neck. As the neck radius vanishes, the governing equations become singular and the filament breaks. In slightly viscous liquids, thinning initially occurs in an inertial regime where inertial and capillary forces balance. By contrast, in highly viscous liquids, initial thinning occurs in a viscous regime where viscous and capillary forces balance. As the filament thins, viscous forcesmore » in the former case and inertial forces in the latter become important, and theory shows that the filament approaches breakup in the final inertial–viscous regime where all three forces balance. However, previous simulations and experiments reveal that transition from an initial to the final regime either occurs at a value of filament radius well below that predicted by theory or is not observed. In this paper, we perform new simulations and experiments, and show that a thinning filament unexpectedly passes through a number of intermediate transient regimes, thereby delaying onset of the inertial–viscous regime. Finally, the new findings have practical implications regarding formation of undesirable satellite droplets and also raise the question as to whether similar dynamical transitions arise in other free-surface flows such as coalescence that also exhibit singularities.« less

Plethora of transitions during breakup of liquid filaments

PubMed Central

Castrejón-Pita, José Rafael; Castrejón-Pita, Alfonso Arturo; Thete, Sumeet Suresh; Sambath, Krishnaraj; Hutchings, Ian M.; Hinch, John; Lister, John R.; Basaran, Osman A.

2015-01-01

Thinning and breakup of liquid filaments are central to dripping of leaky faucets, inkjet drop formation, and raindrop fragmentation. As the filament radius decreases, curvature and capillary pressure, both inversely proportional to radius, increase and fluid is expelled with increasing velocity from the neck. As the neck radius vanishes, the governing equations become singular and the filament breaks. In slightly viscous liquids, thinning initially occurs in an inertial regime where inertial and capillary forces balance. By contrast, in highly viscous liquids, initial thinning occurs in a viscous regime where viscous and capillary forces balance. As the filament thins, viscous forces in the former case and inertial forces in the latter become important, and theory shows that the filament approaches breakup in the final inertial–viscous regime where all three forces balance. However, previous simulations and experiments reveal that transition from an initial to the final regime either occurs at a value of filament radius well below that predicted by theory or is not observed. Here, we perform new simulations and experiments, and show that a thinning filament unexpectedly passes through a number of intermediate transient regimes, thereby delaying onset of the inertial–viscous regime. The new findings have practical implications regarding formation of undesirable satellite droplets and also raise the question as to whether similar dynamical transitions arise in other free-surface flows such as coalescence that also exhibit singularities. PMID:25825761

Increasing cyclic electron flow is related to Na+ sequestration into vacuoles for salt tolerance in soybean.

PubMed

He, Yi; Fu, Junliang; Yu, Chenliang; Wang, Xiaoman; Jiang, Qinsu; Hong, Jian; Lu, Kaixing; Xue, Gangping; Yan, Chengqi; James, Andrew; Xu, Ligen; Chen, Jianping; Jiang, Dean

2015-11-01

In land plants, the NAD(P)H dehydrogenase (NDH) complex reduces plastoquinones and drives cyclic electron flow (CEF) around PSI. It also produces extra ATP for photosynthesis and improves plant fitness under conditions of abiotic environmental stress. To elucidate the role of CEF in salt tolerance of the photosynthetic apparatus, Na(+) concentration, chlorophyll fluorescence, and expression of NDH B and H subunits, as well as of genes related to cellular and vacuolar Na(+) transport, were monitored. The salt-tolerant Glycine max (soybean) variety S111-9 exhibited much higher CEF activity and ATP accumulation in light than did the salt-sensitive variety Melrose, but similar leaf Na(+) concentrations under salt stress. In S111-9 plants, ndhB and ndhH were highly up-regulated under salt stress and their corresponding proteins were maintained at high levels or increased significantly. Under salt stress, S111-9 plants accumulated Na(+) in the vacuole, but Melrose plants accumulated Na(+) in the chloroplast. Compared with Melrose, S111-9 plants also showed higher expression of some genes associated with Na(+) transport into the vacuole and/or cell, such as genes encoding components of the CBL10 (calcineurin B-like protein 10)-CIPK24 (CBL-interacting protein kinase 24)-NHX (Na(+)/H(+) antiporter) and CBL4 (calcineurin B-like protein 4)-CIPK24-SOS1 (salt overly sensitive 1) complexes. Based on the findings, it is proposed that enhanced NDH-dependent CEF supplies extra ATP used to sequester Na(+) in the vacuole. This reveals an important mechanism for salt tolerance in soybean and provides new insights into plant resistance to salt stress. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

SYMPATHETIC SOLAR FILAMENT ERUPTIONS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Rui; Liu, Ying D.; Zimovets, Ivan

2016-08-10

The 2015 March 15 coronal mass ejection as one of the two that together drove the largest geomagnetic storm of solar cycle 24 so far was associated with sympathetic filament eruptions. We investigate the relations between the different filaments involved in the eruption. A surge-like small-scale filament motion is confirmed as the trigger that initiated the erupting filament with multi-wavelength observations and using a forced magnetic field extrapolation method. When the erupting filament moved to an open magnetic field region, it experienced an obvious acceleration process and was accompanied by a C-class flare and the rise of another larger filamentmore » that eventually failed to erupt. We measure the decay index of the background magnetic field, which presents a critical height of 118 Mm. Combining with a potential field source surface extrapolation method, we analyze the distributions of the large-scale magnetic field, which indicates that the open magnetic field region may provide a favorable condition for F2 rapid acceleration and have some relation with the largest solar storm. The comparison between the successful and failed filament eruptions suggests that the confining magnetic field plays an important role in the preconditions for an eruption.« less

The effective resistance between twisted superconducting filaments in tapes

NASA Astrophysics Data System (ADS)

Takács, S.; Iwakuma, M.; Funaki, K.

2001-05-01

We consider two mechanisms, which influence the effective resistance between crossing strands on flat cables or filaments in twisted tapes. The one-layer classical Rutherford-type cable and the tapes with twisted BSCCO filaments in silver matrix are taken as analogous cases. The amount of the matrix between strands or filaments increases the effective conductance compared with the direct current paths (determined by the touching area of the filaments). The increase factor is about two and can easily be suppressed by other effects, like the contact resistance between the superconductor and the matrix. The second mechanism is due to the existence of induced voltage between any points of crossing filaments. This leads to an additional effective conductance, proportional to the square of the total number of filaments. Both effects are not very important for isotropic superconductors, but due to the strong anisotropy of critical parameters they can dominate for high temperature superconductors. The first one may partially compensate the influence of the usually weaker critical current density perpendicular to the tape. The contribution due to the second effect can explain the higher resistivity of the matrix in BSCCO tapes compared with pure silver. It seems that to obtain low AC coupling losses in BSCCO tapes, structures with small filament number are required.

Torins are potent antimalarials that block replenishment of Plasmodium liver stage parasitophorous vacuole membrane proteins

PubMed Central

Hanson, Kirsten K.; Ressurreição, Ana S.; Buchholz, Kathrin; Prudêncio, Miguel; Herman-Ornelas, Jonathan D.; Rebelo, Maria; Beatty, Wandy L.; Wirth, Dyann F.; Hänscheid, Thomas; Moreira, Rui; Marti, Matthias; Mota, Maria M.

2013-01-01

Residence within a customized vacuole is a highly successful strategy used by diverse intracellular microorganisms. The parasitophorous vacuole membrane (PVM) is the critical interface between Plasmodium parasites and their possibly hostile, yet ultimately sustaining, host cell environment. We show that torins, developed as ATP-competitive mammalian target of rapamycin (mTOR) kinase inhibitors, are fast-acting antiplasmodial compounds that unexpectedly target the parasite directly, blocking the dynamic trafficking of the Plasmodium proteins exported protein 1 (EXP1) and upregulated in sporozoites 4 (UIS4) to the liver stage PVM and leading to efficient parasite elimination by the hepatocyte. Torin2 has single-digit, or lower, nanomolar potency in both liver and blood stages of infection in vitro and is likewise effective against both stages in vivo, with a single oral dose sufficient to clear liver stage infection. Parasite elimination and perturbed trafficking of liver stage PVM-resident proteins are both specific aspects of torin-mediated Plasmodium liver stage inhibition, indicating that torins have a distinct mode of action compared with currently used antimalarials. PMID:23836641

Filament cooling and condensation in a sheared magnetic field

NASA Technical Reports Server (NTRS)

Van Hoven, Gerard

1990-01-01

Thermal instability driven by optically thin radiation in the corona is believed to initiate the formation of solar filaments. The fact that filaments are observed generally to separate regions of opposite, line-of-sight, magnetic polarity in the differentially rotating photosphere suggests that filament formation requires the presence of a highly sheared magnetic field. The coupled energetics and dynamics of the most important condensation modes, those due to perpendicular thermal conduction at short wavelengths are discussed. Linear structure in the sheared field and their growth rates is described, and 2D, nonlinear, MHD simulations of the evolution of these modes in a force-free field are conducted. The simulations achieve the fine thermal structures, minimum temperatures and maximum densities characteristic of observed solar filaments.

NGVLA Observations of Dense Gas Filaments in Star-Forming Regions

NASA Astrophysics Data System (ADS)

Di Francesco, James; Chen, Mike; Keown, Jared; GAS Team, KEYSTONE Team

2018-01-01

Recent observations of continuum emission from nearby star-forming regions with Herschel and JCMT have revealed that filaments are ubiquitous structures within molecular clouds. Such filaments appear to be intimately connected to star formation, with those having column densities of AV > 8 hosting the majority of prestellar cores and young protostars in clouds. Indeed, this “threshold” can be explained simply as the result of supercritical cylinder fragmentation. How specifically star-forming filaments form in molecular clouds, however, remains unclear, though gravity and turbulence are likely involved. Observations of their kinematics are needed to understand how mass flows both onto and through these filaments. We show here results from two recent surveys, the Green Bank Ammonia Survey (GAS) and the K-band Examinations of Young Stellar Object Natal Environments (KEYSTONE) that have used the Green Bank Telescope’s K-band Focal Plane Array instrument to map NH3 (1,1) emission from dense gas in nearby star-forming regions. Data from both surveys show that NH3 emission traces extremely well the high column density gas across these star-forming regions. In particular, the GAS results for NGC 1333 show NH3-based velocity gradients either predominantly parallel or perpendicular to the filament spines. Though the GAS and KEYSTONE data are vital for probing filaments, higher resolutions than possible with the GBT alone are needed to examine the kinematic patterns on the 0.1-pc scales of star-forming cores within filaments. We describe how the Next Generation Very Large Array (NGVLA) will uniquely provide the key wide-field data of high sensitivity needed to explore how ambient gas in molecular clouds forms filaments that evolve toward star formation.

Material Supply and Magnetic Configuration of an Active Region Filament

NASA Astrophysics Data System (ADS)

Zou, P.; Fang, C.; Chen, P. F.; Yang, K.; Hao, Q.; Cao, Wenda

2016-11-01

It is important to study the fine structures of solar filaments with high-resolution observations, since it can help us understand the magnetic and thermal structures of the filaments and their dynamics. In this paper, we study a newly formed filament located inside the active region NOAA 11762, which was observed by the 1.6 m New Solar Telescope at Big Bear Solar Observatory from 16:40:19 UT to 17:07:58 UT on 2013 June 5. As revealed by the Hα filtergrams, cool material is seen to be injected into the filament spine with a speed of 5-10 km s-1. At the source of the injection, brightenings are identified in the chromosphere, which are accompanied by magnetic cancellation in the photosphere, implying the importance of magnetic reconnection in replenishing the filament with plasmas from the lower atmosphere. Counter-streamings are detected near one endpoint of the filament, with the plane-of-the-sky speed being 7-9 km s-1 in the Hα red-wing filtergrams and 9-25 km s-1 in the blue-wing filtergrams. The observations are indicative that this active region filament is supported by a sheared arcade without magnetic dips, and the counter-streamings are due to unidirectional flows with alternative directions, rather than due to the longitudinal oscillations of filament threads as in many other filaments.

LAMP proteins account for the maturation delay during the establishment of the Coxiella burnetii-containing vacuole.

PubMed

Schulze-Luehrmann, Jan; Eckart, Rita A; Ölke, Martha; Saftig, Paul; Liebler-Tenorio, Elisabeth; Lührmann, Anja

2016-02-01

The obligate intracellular pathogen Coxiella burnetii replicates in a large phagolysosomal-like vacuole. Currently, both host and bacterial factors required for creating this replicative parasitophorous C. burnetii-containing vacuole (PV) are poorly defined. Here, we assessed the contributions of the most abundant proteins of the lysosomal membrane, LAMP-1 and LAMP-2, to the establishment and maintenance of the PV. Whereas these proteins were not critical for uptake of C. burnetii, they influenced the intracellular replication of C. burnetii. In LAMP-1/2 double-deficient fibroblasts as well as in LAMP-1/2 knock-down cells, C. burnetii establishes a significantly smaller, yet faster maturing vacuole, which harboured more bacteria. The accelerated maturation of PVs in LAMP double-deficient fibroblasts, which was partially or fully reversed by ectopic expression of LAMP-1 or LAMP-2, respectively, was characterized by an increased fusion rate with endosomes, lysosomes and bead-containing phagosomes, but not by different fusion kinetics with autophagy vesicles. These findings establish that LAMP proteins are critical for the maturation delay of PVs. Unexpectedly, neither the creation of the spacious vacuole nor the delay in maturation was found to be prerequisites for the intracellular replication of C. burnetii. © 2015 John Wiley & Sons Ltd.

Influence of various superhydrophilic treatments of titanium on the initial attachment, proliferation, and differentiation of osteoblast-like cells.

PubMed

Yamamura, Keisuke; Miura, Tadashi; Kou, I; Muramatsu, Takashi; Furusawa, Masahiro; Yoshinari, Masao

2015-01-01

The purpose of this study was to investigate the influence of superhydrophilic treatments of titanium on the behavior of osteoblastlike cells. Superhydrophilic specimens were prepared with sandblast and acid-etching (DW), oxygen plasma (Plasma) and ultraviolet light (UV), and were stored in distilled water for 3 days immediately after these treatments. Specimens stored in air for 3 weeks were used as a control Air group. Initial cell attachment, proliferation, alkaline phosphatase activity, and osteocalcin secretion of mouse osteoblast-like cells MC3T3-E1 were enhanced more on superhydrophilic groups than were Air specimens. On confocal laser scanning microscope images of cell morphology, the expression of actin filaments was observed on the superhydrophilic groups, whereas relatively little actin filament expression was seen on the Air surfaces on all culture periods. These results indicate that DW, Plasma, or UV treatment has potential for the creation and maintenance of superhydrophilic surfaces and the enhancement of the initial attachment, proliferation, and differentiation of osteoblast-like cells.

Laser-filamentation-induced condensation and snow formation in a cloud chamber.

PubMed

Ju, Jingjing; Liu, Jiansheng; Wang, Cheng; Sun, Haiyi; Wang, Wentao; Ge, Xiaochun; Li, Chuang; Chin, See Leang; Li, Ruxin; Xu, Zhizhan

2012-04-01

Using 1 kHz, 9 mJ femtosecond laser pulses, we demonstrate laser-filamentation-induced spectacular snow formation in a cloud chamber. An intense updraft of warm moist air is generated owing to the continuous heating by the high-repetition filamentation. As it encounters the cold air above, water condensation and large-sized particles spread unevenly across the whole cloud chamber via convection and cyclone like action on a macroscopic scale. This indicates that high-repetition filamentation plays a significant role in macroscopic laser-induced water condensation and snow formation.

A penny-shaped crack in a filament-reinforced matrix. I - The filament model. II - The crack problem

NASA Technical Reports Server (NTRS)

Erdogan, F.; Pacella, A. H.

1974-01-01

The study deals with the elastostatic problem of a penny-shaped crack in an elastic matrix which is reinforced by filaments or fibers perpendicular to the plane of the crack. An elastic filament model is first developed, followed by consideration of the application of the model to the penny-shaped crack problem in which the filaments of finite length are asymmetrically distributed around the crack. Since the primary interest is in the application of the results to studies relating to the fracture of fiber or filament-reinforced composites and reinforced concrete, the main emphasis of the study is on the evaluation of the stress intensity factor along the periphery of the crack, the stresses in the filaments or fibers, and the interface shear between the matrix and the filaments or fibers. Using the filament model developed, the elastostatic interaction problem between a penny-shaped crack and a slender inclusion or filament in an elastic matrix is formulated.

Tungsten Filament Fire

ERIC Educational Resources Information Center

Ruiz, Michael J.; Perkins, James

2016-01-01

We safely remove the outer glass bulb from an incandescent lamp and burn up the tungsten filament after the glass is removed. This demonstration dramatically illustrates the necessity of a vacuum or inert gas for the environment surrounding the tungsten filament inside the bulb. Our approach has added historical importance since the incandescent…

Influence of KDEL on the fate of trimeric or assembly-defective phaseolin: selective use of an alternative route to vacuoles.

PubMed

Frigerio, L; Pastres, A; Prada, A; Vitale, A

2001-05-01

The tetrapeptide KDEL is commonly found at the C terminus of soluble proteins of the endoplasmic reticulum (ER), and it contributes to their localization by interacting with a receptor that recycles between the Golgi complex and the ER. We investigated the effects of the addition of KDEL to phaseolin, a protein normally delivered from the ER to storage vacuoles via the Golgi complex. We show that KDEL prevents acquisition of trans-Golgi-specific glycan modifications and causes interactions with the chaperone BiP that are distinct from the ones between BiP and defective proteins. KDEL markedly increases the stability of phaseolin, but a small proportion of phaseolin-KDEL slowly reaches the vacuole without undergoing Golgi-mediated glycan modifications, in a process that can be inhibited by brefeldin A but not monensin. Our results indicate that KDEL can operate with high efficiency before proteins can reach the late Golgi cisternae but allows or promotes delivery to vacuoles via an alternative mechanism. However, addition of KDEL does not alter the destiny of an assembly-defective form of phaseolin, suggesting that the plant ER quality control mechanism is dominant over KDEL effects.

Multifractal properties of solar filaments and sunspots numbers

NASA Astrophysics Data System (ADS)

Wu, Nan; Li, Qi-Xiu; Zou, Peng

2015-07-01

We analyze multifractal properties of low (LLSFNs; < 50 °), high (HLSFNs; ⩾ 50 °), full-disk (FDSFNs; 0 ° ˜ 90 °) solar filament numbers (SFNs) and international sunspot numbers (ISNs) by estimating characteristic parameters (α0, Δα , spectrum skewness) of f (α) singularity spectrum. We find that the SFNs and ISNs have multifractal nature. The obtained α0 and Δα indicate that long-term behaviour of the solar filaments is more complex than that of the sunspots and the high-latitude filaments is the most complex in long-term behaviour. The spectrum skewnesses manifest that the ISNs display well symmetrical distribution in singularity strengths, whereas the SFNs are dominated by low singularity strengths, which means that the long-term behaviour of sunspots has homogenous structures and the filaments display averagely small fluctuations in amplitude. To detect the origin of their multifractality, we decompose the raw data of ISNs and SFNs: smoothed data represent ˜11-year cyclic activities and detrended data represent accidental activities. We also calculate their f (α) spectra, respectively. We find that the ˜11-year cyclic activities of filaments and sunspots tend to be a monofractal and display a bit predominance of low singularity strengths. Their accidental activities have the most complex behaviour than the raw and smoothed data. The accidental activities are dominated by high singularity strengths showing averagely large fluctuations in amplitude. Furthermore, multifractal properties from α0 and Δα of the accidental activities have the same features as that of raw data. We think that the ˜11-year periodic activity determines global fluctuations, while the accidental activities rule local complexity.

ngs (notochord granular surface) gene encodes a novel type of intermediate filament family protein essential for notochord maintenance in zebrafish.

PubMed

Tong, Xiangjun; Xia, Zhidan; Zu, Yao; Telfer, Helena; Hu, Jing; Yu, Jingyi; Liu, Huan; Zhang, Quan; Sodmergen; Lin, Shuo; Zhang, Bo

2013-01-25

The notochord is an important organ involved in embryonic patterning and locomotion. In zebrafish, the mature notochord consists of a single stack of fully differentiated, large vacuolated cells called chordocytes, surrounded by a single layer of less differentiated notochordal epithelial cells called chordoblasts. Through genetic analysis of zebrafish lines carrying pseudo-typed retroviral insertions, a mutant exhibiting a defective notochord with a granular appearance was isolated, and the corresponding gene was identified as ngs (notochord granular surface), which was specifically expressed in the notochord. In the mutants, the notochord started to degenerate from 32 hours post-fertilization, and the chordocytes were then gradually replaced by smaller cells derived from chordoblasts. The granular notochord phenotype was alleviated by anesthetizing the mutant embryos with tricaine to prevent muscle contraction and locomotion. Phylogenetic analysis showed that ngs encodes a new type of intermediate filament (IF) family protein, which we named chordostatin based on its function. Under the transmission electron microcopy, bundles of 10-nm-thick IF-like filaments were enriched in the chordocytes of wild-type zebrafish embryos, whereas the chordocytes in ngs mutants lacked IF-like structures. Furthermore, chordostatin-enhanced GFP (EGFP) fusion protein assembled into a filamentous network specifically in chordocytes. Taken together, our work demonstrates that ngs encodes a novel type of IF protein and functions to maintain notochord integrity for larval development and locomotion. Our work sheds light on the mechanisms of notochord structural maintenance, as well as the evolution and biological function of IF family proteins.

ngs (Notochord Granular Surface) Gene Encodes a Novel Type of Intermediate Filament Family Protein Essential for Notochord Maintenance in Zebrafish*

PubMed Central

Tong, Xiangjun; Xia, Zhidan; Zu, Yao; Telfer, Helena; Hu, Jing; Yu, Jingyi; Liu, Huan; Zhang, Quan; Sodmergen; Lin, Shuo; Zhang, Bo

2013-01-01

The notochord is an important organ involved in embryonic patterning and locomotion. In zebrafish, the mature notochord consists of a single stack of fully differentiated, large vacuolated cells called chordocytes, surrounded by a single layer of less differentiated notochordal epithelial cells called chordoblasts. Through genetic analysis of zebrafish lines carrying pseudo-typed retroviral insertions, a mutant exhibiting a defective notochord with a granular appearance was isolated, and the corresponding gene was identified as ngs (notochord granular surface), which was specifically expressed in the notochord. In the mutants, the notochord started to degenerate from 32 hours post-fertilization, and the chordocytes were then gradually replaced by smaller cells derived from chordoblasts. The granular notochord phenotype was alleviated by anesthetizing the mutant embryos with tricaine to prevent muscle contraction and locomotion. Phylogenetic analysis showed that ngs encodes a new type of intermediate filament (IF) family protein, which we named chordostatin based on its function. Under the transmission electron microcopy, bundles of 10-nm-thick IF-like filaments were enriched in the chordocytes of wild-type zebrafish embryos, whereas the chordocytes in ngs mutants lacked IF-like structures. Furthermore, chordostatin-enhanced GFP (EGFP) fusion protein assembled into a filamentous network specifically in chordocytes. Taken together, our work demonstrates that ngs encodes a novel type of IF protein and functions to maintain notochord integrity for larval development and locomotion. Our work sheds light on the mechanisms of notochord structural maintenance, as well as the evolution and biological function of IF family proteins. PMID:23132861

Visualization of structural organization of ventral membranes of sheared-open resorbing osteoclasts attached to apatite pellets.

PubMed

Akisaka, Toshitaka; Yoshida, Atsushi

2015-05-01

Osteoclasts are highly polarized cells from both morphological and functional points of view. Using quick-freeze, rotary-replication methods combined with cell-shearing, we clarified the variability of cytoplasmic surface of the polarized membranes of osteoclasts seeded on apatite. As to the organization of actin filaments and clathrin sheets, we confirmed almost the same ventral membrane specializations of osteoclasts on apatite as seen on glass plates. The organized actin filaments and membrane-associated particles supported the ruffled border membranes. Inside the actin sealing zone, membrane specializations were not always occupied with the ruffled border but also with other types of membranes. Some osteoclasts formed an actin ring but lacked the ruffled border projections. We report a unique and distinctive membrane modification of apatite-attached osteoclasts, i.e., the presence of dense aggregates of membrane-associated particles and related structures not found in the osteoclasts seeded on glass plates. Actin filament polarity in the podosomes was determined by decoration with myosin S1. The actin filament polarity within podosome appears to be oriented predominantly with its barbed ends toward the core, whereas the interconnecting F-actin appears to be mixed oriented. Two different types of clathrin plaques displayed different distributions: clathrin-dependent endocytosis was observed in the ruffled border regions, whereas flat clathrin sheets were found in the leading edge of lamellipodia and near podosomes. The clathrin sheets adhered to the apatite surface tightly on the ventral membranes overlaying the resorption lacunae. All these membrane specializations as mentioned above may indicate the functional variability of osteoclasts seeded on apatite.

Development of a process for producing ribbon shaped filaments. [production of silicon carbide filaments

NASA Technical Reports Server (NTRS)

Debolt, H. E.; Krukonis, V. J.

1973-01-01

Silicon carbide (SiC) ribbon filaments were produced on a carbon ribbon substrate, about 1500 microns (60 mils) wide and 100 microns (4 mils) thick in lengths up to 2 meters (6 ft), and with tensile strengths up to 142 KN/cm sq (206 Ksi). During the course of the study, ribbon filaments of boron were also produced on the carbon ribbon substrate; the boron ribbon produced was extremely fragile. The tensile strength of the SiC ribbon was limited by large growths or flaws caused by anomalies at the substrate surface; these anomalies were either foreign dirt or substrate imperfections or both. Related work carried out on round 100 micron (4 mils) diameter SiC filaments on a 33 micron (1.3 mil) diameter, very smooth carbon monofilament substrate has shown that tensile strengths as high as 551 KN/cm sq (800 Ksi) are obtainable with the SiC-carbon round substrate combination, and indicates that if the ribbon substrate surface and ribbon deposition process can be improved similar strengths can be realizable. Cost analysis shows that 100 micron x 5-10 micron SiC ribbon can be very low cost reinforcement material.

Brownian motion of polyphosphate complexes in yeast vacuoles: characterization by fluorescence microscopy with image analysis.

PubMed

Puchkov, Evgeny O

2010-06-01

In the vacuoles of Saccharomyces cerevisiae yeast cells, vividly moving insoluble polyphosphate complexes (IPCs) <1 microm size, stainable by a fluorescent dye, 4',6-diamidino-2-phenylindole (DAPI), may appear under some growth conditions. The aim of this study was to quantitatively characterize the movement of the IPCs and to evaluate the viscosity in the vacuoles using the obtained data. Studies were conducted on S. cerevisiae cells stained by DAPI and fluorescein isothyocyanate-labelled latex microspheres, using fluorescence microscopy combined with computer image analysis (ImageJ software, NIH, USA). IPC movement was photorecorded and shown to be Brownian motion. On latex microspheres, a methodology was developed for measuring a fluorescing particle's two-dimensional (2D) displacements and its size. In four yeast cells, the 2D displacements and sizes of the IPCs were evaluated. Apparent viscosity values in the vacuoles of the cells, computed by the Einstein-Smoluchowski equation using the obtained data, were found to be 2.16 +/- 0.60, 2.52 +/- 0.63, 3.32 +/- 0.9 and 11.3 +/- 1.7 cP. The first three viscosity values correspond to 30-40% glycerol solutions. The viscosity value of 11.3 +/- 1.7 cP was supposed to be an overestimation, caused by the peculiarities of the vacuole structure and/or volume in this particular cell. This conclusion was supported by the particular quality of the Brownian motion trajectories set in this cell as compared to the other three cells.

Tropomyosin inhibits ADF/cofilin-dependent actin filament dynamics.

PubMed

Ono, Shoichiro; Ono, Kanako

2002-03-18

Tropomyosin binds to actin filaments and is implicated in stabilization of actin cytoskeleton. We examined biochemical and cell biological properties of Caenorhabditis elegans tropomyosin (CeTM) and obtained evidence that CeTM is antagonistic to ADF/cofilin-dependent actin filament dynamics. We purified CeTM, actin, and UNC-60B (a muscle-specific ADF/cofilin isoform), all of which are derived from C. elegans, and showed that CeTM and UNC-60B bound to F-actin in a mutually exclusive manner. CeTM inhibited UNC-60B-induced actin depolymerization and enhancement of actin polymerization. Within isolated native thin filaments, actin and CeTM were detected as major components, whereas UNC-60B was present at a trace amount. Purified UNC-60B was unable to interact with the native thin filaments unless CeTM and other associated proteins were removed by high-salt extraction. Purified CeTM was sufficient to restore the resistance of the salt-extracted filaments from UNC-60B. In muscle cells, CeTM and UNC-60B were localized in different patterns. Suppression of CeTM by RNA interference resulted in disorganized actin filaments and paralyzed worms in wild-type background. However, in an ADF/cofilin mutant background, suppression of CeTM did not worsen actin organization and worm motility. These results suggest that tropomyosin is a physiological inhibitor of ADF/cofilin-dependent actin dynamics.

High-resolution Observations of Flares in an Arch Filament System

NASA Astrophysics Data System (ADS)

Su, Yingna; Liu, Rui; Li, Shangwei; Cao, Wenda; Ahn, Kwangsu; Ji, Haisheng

2018-03-01

We study five sequential solar flares (SOL2015-08-07) occurring in Active Region 12396 observed with the Goode Solar Telescope (GST) at the Big Bear Solar Observatory, complemented by Interface Region Imaging Spectrograph and SDO observations. The main flaring region is an arch filament system (AFS) consisting of multiple bundles of dark filament threads enclosed by semicircular flare ribbons. We study the magnetic configuration and evolution of the active region by constructing coronal magnetic field models based on SDO/HMI magnetograms using two independent methods, i.e., the nonlinear force-free field (NLFFF) extrapolation and the flux rope insertion method. The models consist of multiple flux ropes with mixed signs of helicity, i.e., positive (negative) in the northern (southern) region, which is consistent with the GST observations of multiple filament bundles. The footprints of quasi-separatrix layers (QSLs) derived from the extrapolated NLFFF compare favorably with the observed flare ribbons. An interesting double-ribbon fine structure located at the east border of the AFS is consistent with the fine structure of the QSL’s footprint. Moreover, magnetic field lines traced along the semicircular footprint of a dome-like QSL surrounding the AFS are connected to the regions of significant helicity and Poynting flux injection. The maps of magnetic twist show that positive twist became dominant as time progressed, which is consistent with the injection of positive helicity before the flares. We hence conclude that these circular shaped flares are caused by 3D magnetic reconnection at the QSLs associated with the AFS possessing mixed signs of helicity.

Unwinding motion of a twisted active region filament

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yan, X. L.; Xue, Z. K.; Kong, D. F.

To better understand the structures of active region filaments and the eruption process, we study an active region filament eruption in active region NOAA 11082 in detail on 2010 June 22. Before the filament eruption, the opposite unidirectional material flows appeared in succession along the spine of the filament. The rising of the filament triggered two B-class flares at the upper part of the filament. As the bright material was injected into the filament from the sites of the flares, the filament exhibited a rapid uplift accompanying the counterclockwise rotation of the filament body. From the expansion of the filament,more » we can see that the filament consisted of twisted magnetic field lines. The total twist of the filament is at least 5π obtained by using a time slice method. According to the morphology change during the filament eruption, it is found that the active region filament was a twisted flux rope and its unwinding motion was like a solar tornado. We also find that there was a continuous magnetic helicity injection before and during the filament eruption. It is confirmed that magnetic helicity can be transferred from the photosphere to the filament. Using the extrapolated potential fields, the average decay index of the background magnetic fields over the filament is 0.91. Consequently, these findings imply that the mechanism of solar filament eruption could be due to the kink instability and magnetic helicity accumulation.« less

The Bipolar Filaments Formed by Herpes Simplex Virus Type 1 SSB/Recombination Protein (ICP8) Suggest a Mechanism for DNA Annealing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Makhov, A.M.; Simon, M.; Sen, A.

2009-02-20

Herpes simplex virus type 1 encodes a multifunctional protein, ICP8, which serves both as a single-strand binding protein and as a recombinase, catalyzing reactions involved in replication and recombination of the viral genome. In the presence of divalent ions and at low temperature, previous electron microscopic studies showed that ICP8 will form long left-handed helical filaments. Here, electron microscopic image reconstruction reveals that the filaments are bipolar, with an asymmetric unit containing two subunits of ICP8 that constitute a symmetrical dimer. This organization of the filament has been confirmed using scanning transmission electron microscopy. The pitch of the filaments ismore » {approx} 250 {angstrom}, with {approx} 6.2 dimers per turn. Docking of a crystal structure of ICP8 into the reconstructed filament shows that the C-terminal domain of ICP8, attached to the body of the subunit by a flexible linker containing {approx} 10 residues, is packed into a pocket in the body of a neighboring subunit in the crystal in a similar manner as in the filament. However, the interactions between the large N-terminal domains are quite different in the filament from that observed in the crystal. A previously proposed model for ICP8 binding single-stranded DNA (ssDNA), based upon the crystal structure, leads to a model for a continuous strand of ssDNA near the filament axis. The bipolar nature of the ICP8 filaments means that a second strand of ssDNA would be running through this filament in the opposite orientation, and this provides a potential mechanism for how ICP8 anneals complementary ssDNA into double-stranded DNA, where each strand runs in opposite directions.« less

Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora

PubMed Central

Werner, Antonia; Herzog, Britta; Frey, Stefan; Pöggeler, Stefanie

2016-01-01

In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL) conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Sm)atg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12. PMID:27309377

Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora.

PubMed

Werner, Antonia; Herzog, Britta; Frey, Stefan; Pöggeler, Stefanie

2016-01-01

In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL) conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Sm)atg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12.

The influence of filament temperature and oxygen concentration on tungsten oxide nanostructures by hot filament metal oxide deposition

NASA Astrophysics Data System (ADS)

Lou, J.; Ye, B. J.; Weng, H. M.; Du, H. J.; Wang, Z. B.; Wang, X. P.

2008-08-01

Tungsten oxide (WOx) nanostructures were prepared by a hot filament chemical vapour deposition system and the temperature of the hot tungsten filaments was changed by steps of degrees. The morphology and average growth rate were indicated by scanning electron microscopy which showed that the morphology was highly related to the filament temperature (Tf) and the distance between the filaments and the polished Si (1 0 0) substrates (df). The influence of Tf on the crystalline nature was studied by x-ray diffraction and Raman spectroscopy. The evolution of stoichiometry and types of defects was indicated by x-ray photoelectron spectroscopy and slow positron implantation spectroscopy. When Tf was up to 1750 °C, tungsten oxide nanostructure was synthesized. A turning point of Tf was found at which the nature of crystallinity and of stoichiometry was the best. As Tf increased to 2100 °C or df decreased, the film crystallinity decreased; correspondingly, the component ratio of stoichiometry WO3 decreased and lots of vacancy agglomerates were present. In order to develop the chemical phase from substoichiometry to stoichiometry, the oxygen gas concentration in the mixture gas during deposition should be raised to an appropriate level.

Laser chirp effect on femtosecond laser filamentation generated for pulse compression.

PubMed

Park, Juyun; Lee, Jae-Hwan; Nam, Chang H

2008-03-31

The influence of laser chirp on the formation of femtosecond laser filamentation in Ar was investigated for the generation of few-cycle high-power laser pulses. The condition for the formation of a single filament has been carefully examined using 28-fs laser pulses with energy over 3 mJ. The filament formation and output spectrum changed very sensitively to the initial laser chirp and gas pressure. Much larger spectral broadening was obtained with positively chirped pulses, compared to the case of negatively chirped pulses that generated much longer filament, and compressed pulses of 5.5 fs with energy of 0.5 mJ were obtained from the filamentation of positively chirped 30-fs laser pulses in a single Ar cell.

Purification and proteomics of pathogen-modified vacuoles and membranes

PubMed Central

Herweg, Jo-Ana; Hansmeier, Nicole; Otto, Andreas; Geffken, Anna C.; Subbarayal, Prema; Prusty, Bhupesh K.; Becher, Dörte; Hensel, Michael; Schaible, Ulrich E.; Rudel, Thomas; Hilbi, Hubert

2015-01-01

Certain pathogenic bacteria adopt an intracellular lifestyle and proliferate in eukaryotic host cells. The intracellular niche protects the bacteria from cellular and humoral components of the mammalian immune system, and at the same time, allows the bacteria to gain access to otherwise restricted nutrient sources. Yet, intracellular protection and access to nutrients comes with a price, i.e., the bacteria need to overcome cell-autonomous defense mechanisms, such as the bactericidal endocytic pathway. While a few bacteria rupture the early phagosome and escape into the host cytoplasm, most intracellular pathogens form a distinct, degradation-resistant and replication-permissive membranous compartment. Intracellular bacteria that form unique pathogen vacuoles include Legionella, Mycobacterium, Chlamydia, Simkania, and Salmonella species. In order to understand the formation of these pathogen niches on a global scale and in a comprehensive and quantitative manner, an inventory of compartment-associated host factors is required. To this end, the intact pathogen compartments need to be isolated, purified and biochemically characterized. Here, we review recent progress on the isolation and purification of pathogen-modified vacuoles and membranes, as well as their proteomic characterization by mass spectrometry and different validation approaches. These studies provide the basis for further investigations on the specific mechanisms of pathogen-driven compartment formation. PMID:26082896

Dry soldering with hot filament produced atomic hydrogen

DOEpatents

Panitz, J.K.G.; Jellison, J.L.; Staley, D.J.

1995-04-25

A system is disclosed for chemically transforming metal surface oxides to metal that is especially, but not exclusively, suitable for preparing metal surfaces for dry soldering and solder reflow processes. The system employs one or more hot, refractory metal filaments, grids or surfaces to thermally dissociate molecular species in a low pressure of working gas such as a hydrogen-containing gas to produce reactive species in a reactive plasma that can chemically reduce metal oxides and form volatile compounds that are removed in the working gas flow. Dry soldering and solder reflow processes are especially applicable to the manufacture of printed circuit boards, semiconductor chip lead attachment and packaging multichip modules. The system can be retrofitted onto existing metal treatment ovens, furnaces, welding systems and wave soldering system designs. 1 fig.

The Pathogen-Occupied Vacuoles of Anaplasma phagocytophilum and Anaplasma marginale Interact with the Endoplasmic Reticulum

PubMed Central

Truchan, Hilary K.; Cockburn, Chelsea L.; Hebert, Kathryn S.; Magunda, Forgivemore; Noh, Susan M.; Carlyon, Jason A.

2016-01-01

The genus Anaplasma consists of tick-transmitted obligate intracellular bacteria that invade white or red blood cells to cause debilitating and potentially fatal infections. A. phagocytophilum, a human and veterinary pathogen, infects neutrophils to cause granulocytic anaplasmosis. A. marginale invades bovine erythrocytes. Evidence suggests that both species may also infect endothelial cells in vivo. In mammalian and arthropod host cells, A. phagocytophilum and A. marginale reside in host cell derived pathogen-occupied vacuoles (POVs). While it was recently demonstrated that the A. phagocytophilum-occupied vacuole (ApV) intercepts membrane traffic from the trans-Golgi network, it is unclear if it or the A. marginale-occupied vacuole (AmV) interacts with other secretory organelles. Here, we demonstrate that the ApV and AmV extensively interact with the host endoplasmic reticulum (ER) in endothelial, myeloid, and/or tick cells. ER lumen markers, calreticulin, and protein disulfide isomerase, and the ER membrane marker, derlin-1, were pronouncedly recruited to the peripheries of both POVs. ApV association with the ER initiated early and continued throughout the infection cycle. Both the ApV and AmV interacted with the rough ER and smooth ER. However, only derlin-1-positive rough ER derived vesicles were delivered into the ApV lumen where they localized with intravacuolar bacteria. Transmission electron microscopy identified multiple ER-POV membrane contact sites on the cytosolic faces of both species' vacuoles that corresponded to areas on the vacuoles' lumenal faces where intravacuolar Anaplasma organisms closely associated. A. phagocytophilum is known to hijack Rab10, a GTPase that regulates ER dynamics and morphology. Yet, ApV-ER interactions were unhindered in cells in which Rab10 had been knocked down, demonstrating that the GTPase is dispensable for the bacterium to parasitize the ER. These data establish the ApV and AmV as pathogen-host interfaces that directly

Emergence of antibiotic resistance from multinucleated bacterial filaments

PubMed Central

Bos, Julia; Zhang, Qiucen; Vyawahare, Saurabh; Rogers, Elizabeth; Rosenberg, Susan M.; Austin, Robert H.

2015-01-01

Bacteria can rapidly evolve resistance to antibiotics via the SOS response, a state of high-activity DNA repair and mutagenesis. We explore here the first steps of this evolution in the bacterium Escherichia coli. Induction of the SOS response by the genotoxic antibiotic ciprofloxacin changes the E. coli rod shape into multichromosome-containing filaments. We show that at subminimal inhibitory concentrations of ciprofloxacin the bacterial filament divides asymmetrically repeatedly at the tip. Chromosome-containing buds are made that, if resistant, propagate nonfilamenting progeny with enhanced resistance to ciprofloxacin as the parent filament dies. We propose that the multinucleated filament creates an environmental niche where evolution can proceed via generation of improved mutant chromosomes due to the mutagenic SOS response and possible recombination of the new alleles between chromosomes. Our data provide a better understanding of the processes underlying the origin of resistance at the single-cell level and suggest an analogous role to the eukaryotic aneuploidy condition in cancer. PMID:25492931

Autophagy-related direct membrane import from ER/cytoplasm into the vacuole or apoplast: a hidden gateway also for secondary metabolites and phytohormones?

PubMed

Kulich, Ivan; Žárský, Viktor

2014-04-29

Transportation of low molecular weight cargoes into the plant vacuole represents an essential plant cell function. Several lines of evidence indicate that autophagy-related direct endoplasmic reticulum (ER) to vacuole (and also, apoplast) transport plays here a more general role than expected. This route is regulated by autophagy proteins, including recently discovered involvement of the exocyst subcomplex. Traffic from ER into the vacuole bypassing Golgi apparatus (GA) acts not only in stress-related cytoplasm recycling or detoxification, but also in developmentally-regulated biopolymer and secondary metabolite import into the vacuole (or apoplast), exemplified by storage proteins and anthocyanins. We propose that this pathway is relevant also for some phytohormones' (e.g., auxin, abscisic acid (ABA) and salicylic acid (SA)) degradation. We hypothesize that SA is not only an autophagy inducer, but also a cargo for autophagy-related ER to vacuole membrane container delivery and catabolism. ER membrane localized enzymes will potentially enhance the area of biosynthetic reactive surfaces, and also, abundant ER localized membrane importers (e.g., ABC transporters) will internalize specific molecular species into the autophagosome biogenesis domain of ER. Such active ER domains may create tubular invaginations of tonoplast into the vacuoles as import intermediates. Packaging of cargos into the ER-derived autophagosome-like containers might be an important mechanism of vacuole and exosome biogenesis and cytoplasm protection against toxic metabolites. A new perspective on metabolic transformations intimately linked to membrane trafficking in plants is emerging.

Autophagy-Related Direct Membrane Import from ER/Cytoplasm into the Vacuole or Apoplast: A Hidden Gateway also for Secondary Metabolites and Phytohormones?

PubMed Central

Kulich, Ivan; Žárský, Viktor

2014-01-01

Transportation of low molecular weight cargoes into the plant vacuole represents an essential plant cell function. Several lines of evidence indicate that autophagy-related direct endoplasmic reticulum (ER) to vacuole (and also, apoplast) transport plays here a more general role than expected. This route is regulated by autophagy proteins, including recently discovered involvement of the exocyst subcomplex. Traffic from ER into the vacuole bypassing Golgi apparatus (GA) acts not only in stress-related cytoplasm recycling or detoxification, but also in developmentally-regulated biopolymer and secondary metabolite import into the vacuole (or apoplast), exemplified by storage proteins and anthocyanins. We propose that this pathway is relevant also for some phytohormones’ (e.g., auxin, abscisic acid (ABA) and salicylic acid (SA)) degradation. We hypothesize that SA is not only an autophagy inducer, but also a cargo for autophagy-related ER to vacuole membrane container delivery and catabolism. ER membrane localized enzymes will potentially enhance the area of biosynthetic reactive surfaces, and also, abundant ER localized membrane importers (e.g., ABC transporters) will internalize specific molecular species into the autophagosome biogenesis domain of ER. Such active ER domains may create tubular invaginations of tonoplast into the vacuoles as import intermediates. Packaging of cargos into the ER-derived autophagosome-like containers might be an important mechanism of vacuole and exosome biogenesis and cytoplasm protection against toxic metabolites. A new perspective on metabolic transformations intimately linked to membrane trafficking in plants is emerging. PMID:24786101

Myosin binding protein-C activates thin filaments and inhibits thick filaments in heart muscle cells

PubMed Central

Kampourakis, Thomas; Yan, Ziqian; Gautel, Mathias; Sun, Yin-Biao; Irving, Malcolm

2014-01-01

Myosin binding protein-C (MyBP-C) is a key regulatory protein in heart muscle, and mutations in the MYBPC3 gene are frequently associated with cardiomyopathy. However, the mechanism of action of MyBP-C remains poorly understood, and both activating and inhibitory effects of MyBP-C on contractility have been reported. To clarify the function of the regulatory N-terminal domains of MyBP-C, we determined their effects on the structure of thick (myosin-containing) and thin (actin-containing) filaments in intact sarcomeres of heart muscle. We used fluorescent probes on troponin C in the thin filaments and on myosin regulatory light chain in the thick filaments to monitor structural changes associated with activation of demembranated trabeculae from rat ventricle by the C1mC2 region of rat MyBP-C. C1mC2 induced larger structural changes in thin filaments than calcium activation, and these were still present when active force was blocked with blebbistatin, showing that C1mC2 directly activates the thin filaments. In contrast, structural changes in thick filaments induced by C1mC2 were smaller than those associated with calcium activation and were abolished or reversed by blebbistatin. Low concentrations of C1mC2 did not affect resting force but increased calcium sensitivity and reduced cooperativity of force and structural changes in both thin and thick filaments. These results show that the N-terminal region of MyBP-C stabilizes the ON state of thin filaments and the OFF state of thick filaments and lead to a novel hypothesis for the physiological role of MyBP-C in the regulation of cardiac contractility. PMID:25512492

Myosin binding protein-C activates thin filaments and inhibits thick filaments in heart muscle cells.

PubMed

Kampourakis, Thomas; Yan, Ziqian; Gautel, Mathias; Sun, Yin-Biao; Irving, Malcolm

2014-12-30

Myosin binding protein-C (MyBP-C) is a key regulatory protein in heart muscle, and mutations in the MYBPC3 gene are frequently associated with cardiomyopathy. However, the mechanism of action of MyBP-C remains poorly understood, and both activating and inhibitory effects of MyBP-C on contractility have been reported. To clarify the function of the regulatory N-terminal domains of MyBP-C, we determined their effects on the structure of thick (myosin-containing) and thin (actin-containing) filaments in intact sarcomeres of heart muscle. We used fluorescent probes on troponin C in the thin filaments and on myosin regulatory light chain in the thick filaments to monitor structural changes associated with activation of demembranated trabeculae from rat ventricle by the C1mC2 region of rat MyBP-C. C1mC2 induced larger structural changes in thin filaments than calcium activation, and these were still present when active force was blocked with blebbistatin, showing that C1mC2 directly activates the thin filaments. In contrast, structural changes in thick filaments induced by C1mC2 were smaller than those associated with calcium activation and were abolished or reversed by blebbistatin. Low concentrations of C1mC2 did not affect resting force but increased calcium sensitivity and reduced cooperativity of force and structural changes in both thin and thick filaments. These results show that the N-terminal region of MyBP-C stabilizes the ON state of thin filaments and the OFF state of thick filaments and lead to a novel hypothesis for the physiological role of MyBP-C in the regulation of cardiac contractility.

Fabrication of Polylactide Nanocomposite Filament Using Melt Extrusion and Filament Characterization for 3D Printing

NASA Astrophysics Data System (ADS)

Jain, Shrenik Kumar

Fused deposition modeling (FDM) technology uses thermoplastic filament for layer by layer fabrication of objects. To make functional objects with desired properties, composite filaments are required in the FDM. In this thesis, less expensive mesoporous Nano carbon (NC) and carbon nanotube (CNT) infused in Polylactide (PLA) thermoplastic filaments were fabricated to improve the electrical properties and maintain sufficient strength for 3D printing. Solution blending was used for nanocomposite fabrication and melt extrusion was employed to make cylindrical filaments. Mechanical and electrical properties of 1 to 20 wt% of NC and 1 to 3 wt% of CNT filaments were investigated and significant improvement of conductivity (3.76 S/m) and sufficient yield strength (35MPa) were obtained. Scanning electron microscopy (SEM) images exhibited uniform dispersion of nanoparticles in polymer matrix and differential scanning calorimetry (DSC) results showed no significant changes in the glass transition temperature (Tg) for all the compositions. Perspective uses of this filament are for fabrication of electrical wires in 3D printed robots, drones, prosthetics, orthotics and others.

Attachment style and suicide behaviors in high risk psychiatric inpatients following hospital discharge: The mediating role of entrapment.

PubMed

Li, Shuang; Galynker, Igor I; Briggs, Jessica; Duffy, Molly; Frechette-Hagan, Anna; Kim, Hae-Joon; Cohen, Lisa J; Yaseen, Zimri S

2017-11-01

Insecure attachment is associated with suicidal behavior. This relationship and its possible mediators have not been examined in high-risk psychiatric inpatients with respect to the critical high-risk period following hospital discharge. Attachment styles and perception of entrapment were assessed in 200 high-risk adult psychiatric inpatients hospitalized following suicidal ideation or suicide attempt. Suicidal behaviors were evaluated with the Columbia Suicide Severity Rating Scale at 1-2 months post-discharge. Associations between different attachment styles and suicidal behaviors were assessed and mediation of attachment effects by entrapment was modeled. Fearful attachment was associated with post-discharge suicidal behavior and there was a trend-level negative association for secure attachment. In addition, entrapment mediated the relationship between fearful attachment and suicidal behavior. The current study highlights the mediating role of perceptions of entrapment in the contribution of fearful attachment to suicidal behavior in high-risk patients, suggesting entrapment as potential therapeutic target to prevent suicidal behavior in these individuals. Further research is warranted to establish the mechanisms by which entrapment experiences emerge in patients with insecure attachment styles. Copyright © 2017 Elsevier B.V. All rights reserved.

UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase in nuclei and rimmed vacuoles of muscle fibers in DMRV (distal myopathy with rimmed vacuoles).

PubMed

Ishihara, Shoichiro; Tomimitsu, Hiroyuki; Fujigasaki, Hiroto; Saito, Fumiaki; Mizusawa, Hidehiro

2008-03-01

UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) is a key molecule in the pathogenesis of distal myopathy with rimmed vacuoles (DMRV) and hereditary inclusion body myopathy (HIBM) and almost all such patients have some mutations in GNE. However, subcellular localization of GNE and the mechanism of muscular damage have not been clarified. A rabbit polyclonal antibody for GNE was prepared. Immunohistochemistry was performed using anti-GNE and anti-nuclear protein antibodies. Western blotting with subcellular fractionated proteins was performed to determine subcellular localization of GNE. The sizes of myonuclei were quantified in muscle biopsies from patients with DMRV and amyotrophic lateral sclerosis (ALS). In DMRV muscles, immunohistochemistry identified GNE in sarcoplasm and specifically in myonuclei and rimmed vacuoles (RV). Nuclear proteins were also found in RVs. Immunohistochemistry showed colocalization of GNE and emerin in C2C12 cells. Western blotting revealed the presence of GNE in nuclear fractions of human embryonic kidney (HEK) 293T cells. The mean size of myonuclei of DMRV was significantly larger than that of ALS. GNE is present in myonuclei near nuclear membrane. Our results suggest that myonuclei are involved in RV formation in DMRV, and that mutant GNE in myonuclei seems to play some role in this process.

High frame-rate resolution of cell division during Candida albicans filamentation

PubMed Central

Thomson, Darren D.; Berman, Judith; Brand, Alexandra C.

2016-01-01

The commensal yeast, Candida albicans, is an opportunistic pathogen in humans and forms filaments called hyphae and pseudohyphae, in which cell division requires precise temporal and spatial control to produce mononuclear cell compartments. High-frame-rate live-cell imaging (1 frame/min) revealed that nuclear division did not occur across the septal plane. We detected the presence of nucleolar fragments that may be extrachromosomal molecules carrying the ribosomal RNA genes. Cells occasionally maintained multiple nucleoli, suggesting either polyploidy, multiple nuclei and/or aneuploidy of ChrR., while the migration pattern of sister nuclei differed between unbranched and branched hyphae. The presented movie challenges and extends previous concepts of C. albicans cell division. PMID:26854071

Current Progress in Tonoplast Proteomics Reveals Insights into the Function of the Large Central Vacuole

PubMed Central

Trentmann, Oliver; Haferkamp, Ilka

2013-01-01

Vacuoles of plants fulfill various biologically important functions, like turgor generation and maintenance, detoxification, solute sequestration, or protein storage. Different types of plant vacuoles (lytic versus protein storage) are characterized by different functional properties apparently caused by a different composition/abundance and regulation of transport proteins in the surrounding membrane, the tonoplast. Proteome analyses allow the identification of vacuolar proteins and provide an informative basis for assigning observed transport processes to specific carriers or channels. This review summarizes techniques required for vacuolar proteome analyses, like e.g., isolation of the large central vacuole or tonoplast membrane purification. Moreover, an overview about diverse published vacuolar proteome studies is provided. It becomes evident that qualitative proteomes from different plant species represent just the tip of the iceberg. During the past few years, mass spectrometry achieved immense improvement concerning its accuracy, sensitivity, and application. As a consequence, modern tonoplast proteome approaches are suited for detecting alterations in membrane protein abundance in response to changing environmental/physiological conditions and help to clarify the regulation of tonoplast transport processes. PMID:23459586

Filament organization revealed in platinum replicas of freeze-dried cytoskeletons

PubMed Central

1980-01-01

This report presents the appearance of rapidly frozen, freeze-dried cytoskeletons that have been rotary replicated with platinum and viewed in the transmission electron microscope. The resolution of this method is sufficient to visualize individual filaments in the cytoskeleton and to discriminate among actin, microtubules, and intermediate filaments solely by their surface substructure. This identification has been confirmed by specific decoration with antibodies and selective extraction of individual filament types, and correlated with light microscope immunocytochemistry and gel electrophoresis patterns. The freeze-drying preserves a remarkable degree of three-dimensionality in the organization of these cytoskeletons. They look strikingly similar to the meshwork of strands or "microtrabeculae" seen in the cytoplasm of whole cells by high voltage electron microscopy, in that the filaments form a lattice of the same configutation and with the same proportions of open area as the microtrabeculae seen in whole cells. The major differences between these two views of the structural elements of the cytoplasmic matrix can be attributed to the effects of aldehyde fixation and dehydration. Freeze-dried cytoskeletons thus provide an opportunity to study--at high resolution and in the absence of problems caused by chemical fixation--the detailed organization of filaments in different regions of the cytoplasm and at different stages of cell development. In this report the pattern of actin and intermediate filament organization in various regions of fully spread mouse fibroblasts is described. PMID:6893451

Conformational phases of membrane bound cytoskeletal filaments

NASA Astrophysics Data System (ADS)

Quint, David A.; Grason, Gregory; Gopinathan, Ajay

2013-03-01

Membrane bound cytoskeletal filaments found in living cells are employed to carry out many types of activities including cellular division, rigidity and transport. When these biopolymers are bound to a membrane surface they may take on highly non-trivial conformations as compared to when they are not bound. This leads to the natural question; What are the important interactions which drive these polymers to particular conformations when they are bound to a surface? Assuming that there are binding domains along the polymer which follow a periodic helical structure set by the natural monomeric handedness, these bound conformations must arise from the interplay of the intrinsic monomeric helicity and membrane binding. To probe this question, we study a continuous model of an elastic filament with intrinsic helicity and map out the conformational phases of this filament for various mechanical and structural parameters in our model, such as elastic stiffness and intrinsic twist of the filament. Our model allows us to gain insight into the possible mechanisms which drive real biopolymers such as actin and tubulin in eukaryotes and their prokaryotic cousins MreB and FtsZ to take on their functional conformations within living cells.

Controlling Plasma Channels through Ultrashort Laser Pulse Filamentation

NASA Astrophysics Data System (ADS)

Ionin, Andrey; Seleznev, Leonid; Sunchugasheva, Elena

2013-09-01

A review of studies fulfilled at the Lebedev Institute in collaboration with the Moscow State University and Institute of Atmospheric Optics in Tomsk on influence of various characteristics of ultrashort laser pulse on plasma channels formed under its filamentation is presented. Filamentation of high-power laser pulses with wavefront controlled by a deformable mirror, with cross-sections spatially formed by various diaphragms and with different wavelengths was experimentally and numerically studied. An application of plasma channels formed due to filamentation of ultrashort laser pulse including a train of such pulses for triggering and guiding long electric discharges is discussed. The research was supported by RFBR Grants 11-02-12061-ofi-m and 11-02-01100, and EOARD Grant 097007 through ISTC Project 4073 P

Ultrastructural characteristics of tau filaments in tauopathies: immuno-electron microscopic demonstration of tau filaments in tauopathies.

PubMed

Arima, Kunimasa

2006-10-01

The microtubule-associated protein tau aggregates into filaments in the form of neurofibrillary tangles, neuropil threads and argyrophilic grains in neurons, in the form of variable astrocytic tangles in astrocytes and in the form of coiled bodies and argyrophilic threads in oligodendrocytes. These tau filaments may be classified into two types, straight filaments or tubules with 9-18 nm diameters and "twisted ribbons" composed of two parallel aligned components. In the same disease, the fine structure of tau filaments in glial cells roughly resembles that in neurons. In sporadic tauopathies, individual tau filaments show characteristic sizes, shapes and arrangements, and therefore contribute to neuropathologic differential diagnosis. In frontotemporal dementias caused by tau gene mutations, variable filamentous profiles were observed in association with mutation sites and insoluble tau isoforms, including straight filaments or tubules, paired helical filament-like filaments, and twisted ribbons. Pre-embedding immunoelectron microscopic studies were carried out using anti-3-repeat tau and anti-4-repeat tau specific antibodies, RD3 and RD4. Straight tubules in neuronal and astrocytic Pick bodies were immunolabeled by the anti-3-repeat tau antibody. The anti-4-repeat tau antibody recognized abnormal tubules comprising neurofibrillary tangles, coiled bodies and argyrophilic threads in progressive supranuclear palsy (PSP) and corticobasal degeneration. In the pre-embedding immunoelectron microscopic study using the phosphorylated tau AT8 antibody, tuft-shaped astrocytes of PSP were found to be composed of bundles of abnormal tubules in processes and perikarya of protoplasmic astrocytes. In this study, the 3-repeat tau or 4-repeat tau epitope was detected in situ at the ultrastructural level in abnormal tubules in representative pathological lesions in Pick's disease, PSP and corticobasal degeneration.

Fast deuterium fractionation in magnetized and turbulent filaments

NASA Astrophysics Data System (ADS)

Körtgen, B.; Bovino, S.; Schleicher, D. R. G.; Stutz, A.; Banerjee, R.; Giannetti, A.; Leurini, S.

2018-04-01

Deuterium fractionation is considered as an important process to infer the chemical ages of prestellar cores in filaments. We present here the first magneto-hydrodynamical simulations including a chemical network to study deuterium fractionation in magnetized and turbulent filaments, with a line-mass of Mlin = 42 M⊙ pc-1 within a radius of R = 0.1 pc, and their substructures. The filaments typically show widespread deuterium fractionation with average values ≳ 0.01. For individual cores of similar age, we observe the deuteration fraction to increase with time, but also to be independent of their average properties such as density, virial or mass-to-magnetic flux ratio. We further find a correlation of the deuteration fraction with core mass, average H2 density and virial parameter only at late evolutionary stages of the filament and attribute this to the lifetime of the individual cores. Specifically, chemically old cores reveal higher deuteration fractions. Within the radial profiles of selected cores, we notice differences in the structure of the deuteration fraction or surface density, which we can attribute to their different turbulent properties. High deuteration fractions of the order 0.01 - 0.1 may be reached within approximately 200 kyrs, corresponding to two free-fall times, as defined for cylindrical systems, of the filaments.

Fast deuterium fractionation in magnetized and turbulent filaments

NASA Astrophysics Data System (ADS)

Körtgen, B.; Bovino, S.; Schleicher, D. R. G.; Stutz, A.; Banerjee, R.; Giannetti, A.; Leurini, S.

2018-07-01

Deuterium fractionation is considered as an important process to infer the chemical ages of prestellar cores in filaments. We present here the first magnetohydrodynamical simulations including a chemical network to study deuterium fractionation in magnetized and turbulent filaments, with a line-mass of Mlin = 42 M⊙ pc-1 within a radius of R= 0.1 pc, and their sub-structures. The filaments typically show widespread deuterium fractionation with average values ≳0.01. For individual cores of similar age, we observe the deuteration fraction to increase with time, but also to be independent of their average properties such as density, virial, or mass-to-magnetic flux ratio. We further find a correlation of the deuteration fraction with core mass, average H2 density, and virial parameter only at late evolutionary stages of the filament and attribute this to the lifetime of the individual cores. Specifically, chemically old cores reveal higher deuteration fractions. Within the radial profiles of selected cores, we notice differences in the structure of the deuteration fraction or surface density, which we can attribute to their different turbulent properties. High deuteration fractions of the order of 0.01-0.1 may be reached within approximately 200 kyr, corresponding to two free-fall times, as defined for cylindrical systems, of the filaments.

Attachment techniques for high temperature strain

NASA Astrophysics Data System (ADS)

Wnuk, Steve P., Jr.

1993-01-01

Attachment methods for making resistive strain measurements to 2500 F were studied. A survey of available strain gages and attachment techniques was made, and the results are compiled for metal and carbon composite test materials. A theoretical analysis of strain transfer into a bonded strain gage was made, and the important physical parameters of the strain transfer medium, the ceramic matrix, were identified. A pull tester to measure pull-out tests on commonly used strain gage cements indicated that all cements tested displayed adequate strength for good strain transfer. Rokide flame sprayed coatings produced significantly stronger bonds than ceramic cements. An in-depth study of the flame spray process produced simplified installation procedures which also resulted in greater reliability and durability. Application procedures incorporating improvements made during this program are appended to the report. Strain gages installed on carbon composites, Rene' 41, 316 stainless steel, and TZM using attachment techniques developed during this program were successfully tested to 2500 F. Photographs of installation techniques, test procedures, and graphs of the test data are included in this report.

A hot X-ray filament associated with A3017 galaxy cluster

NASA Astrophysics Data System (ADS)

Parekh, V.; Durret, F.; Padmanabh, P.; Pandge, M. B.

2017-09-01

Recent simulations and observations have shown large-scale filaments in the cosmic web connecting nodes, with accreting materials (baryonic and dark matter) flowing through them. Current high-sensitivity observations also show that the propagation of shocks through filaments can heat them up and make filaments visible between two or more galaxy clusters or around massive clusters, based on optical and/or X-ray observations. We are reporting here the special case of the cluster A3017 associated with a hot filament. The temperature of the filament is 3.4^{-0.77}_{+1.30} keV and its length is ∼1 Mpc. We have analysed its archival Chandra data and report various properties. We also analysed GMRT 235/610 MHz radio data. Radio observations have revealed symmetric two-sided lobes that fill cavities in the A3017 cluster core region, associated with central active galactic nucleus. In the radio map, we also noticed a peculiar linear vertical radio structure in the X-ray filament region which might be associated with a cosmic filament shock. This radio structure could be a radio phoenix or old plasma where an old relativistic population is re-accelerated by shock propagation. Finally, we put an upper limit on the radio luminosity of the filament region.

Additive Manufacturing of Syntactic Foams: Part 1: Development, Properties, and Recycling Potential of Filaments

NASA Astrophysics Data System (ADS)

Singh, Ashish Kumar; Patil, Balu; Hoffmann, Niklas; Saltonstall, Brooks; Doddamani, Mrityunjay; Gupta, Nikhil

2018-03-01

This work focuses on developing filaments of high-density polyethylene (HDPE) and their hollow particle-filled syntactic foams for commercial three-dimensional (3D) printers based on fused filament fabrication technology. Hollow fly-ash cenospheres were blended by 40 wt.% in a HDPE matrix to produce syntactic foam (HDPE40) filaments. Further, the recycling potential was studied by pelletizing the filaments again to extrude twice (2×) and three times (3×). The filaments were tensile tested at 10-4 s-1, 10-3 s-1, and 10-2 s-1 strain rates. HDPE40 filaments show an increasing trend in modulus and strength with the strain rate. Higher density and modulus were noticed for 2× filaments compared to 1× filaments because of the crushing of some cenospheres in the extrusion cycle. However, 2× and 3× filament densities are nearly the same, showing potential for recycling them. The filaments show better properties than the same materials processed by conventional injection molding. Micro-CT scans show a uniform dispersion of cenospheres in all filaments.

Transmission electron microscopy of the preclinical phase of experimental phytophotodermatitis.

PubMed

Almeida, Hiram Larangeira de; Sotto, Miriam Nakagami; Castro, Luis Antonio Suita de; Rocha, Nara Moreira

2008-06-01

To examine the epidermis in induced phytophotodermatitis using transmission electron microscopy in order to detect histologic changes even before lesions are visible by light microscopy. In the first six hours after the experimental induction of phytophotodermatitis, no changes are detectable by light microscopy. Only after 24 hours can keratinocyte necrosis and epidermal vacuolization be detected histologically, and blisters form by 48 hours. The dorsum of four adult rats (Rattus norvegicus) was manually epilated. After painting the right half of the rat with the peel juice of Tahiti lemon, they were exposed to sunlight for eight minutes under general anesthesia. The left side was used as the control and exposed to sunlight only. Biopsies were performed immediately after photoinduction and one and two hours later, and the tissue was analyzed by transmission electron microscopy. No histological changes were seen on the control side. Immediately after induction, vacuolization in keratinocytes was observed. After one hour, desmosomal changes were also observed in addition to vacuolization. Keratin filaments were not attached to the desmosomal plaque. Free desmosomes and membrane ruptures were also seen. At two hours after induction, similar changes were found, and granular degeneration of keratin was also observed. The interaction of sunlight and psoralens generates a photoproduct that damages keratinocyte proteins, leading to keratinocyte necrosis and blister formation. Transmission electron microscopy can detect vacuolization, lesions of the membrane, and desmosomes in the first two hours after experimental induction of phytophotodermatitis.

Light microscopy and image analysis of thin filament lengths utilizing dual probes on beef, chicken, and rabbit myofibrils.

PubMed

Ringkob, T P; Swartz, D R; Greaser, M L

2004-05-01

Image analysis procedures for immunofluorescence microscopy were developed to measure muscle thin filament lengths of beef, rabbit, and chicken myofibrils. Strips of beef cutaneous trunci, rectus abdominis, psoas, and masseter; chicken pectoralis; and rabbit psoas muscles were excised 5 to 30 min postmortem. Fluorescein phalloidin and rhodamine myosin subfragment-1 (S1) were used to probe the myofibril structure. Digital images were recorded with a cooled charge-coupled device controlled with IPLab Spectrum software (Signal Analytics Corp.) on a Macintosh operating system. The camera was attached to an inverted microscope, using both the phase-contrast and fluorescence illumination modes. Unfixed myofibrils incubated with fluorescein phalloidin showed fluorescence primarily at the Z-line and the tips of the thin filaments in the overlap region. Images were processed using IPLab and the National Institutes of Health's Image software. A region of interest was selected and scaled by a factor of 18.18, which enlarged the image from 11 pixels/microm to approximately 200 pixels/microm. An X-Y plot was exported to Spectrum 1.1 (Academic Software Development Group), where the signal was processed with a second derivative routine, so a cursor function could be used to measure length. Fixation before phalloidin incubation resulted in greatest intensity at the Z lines but a more-uniform staining over the remainder of the thin filament zone. High-resolution image capture and processing showed that thin filament lengths were significantly different (P < 0.01) among beef, rabbit, and chicken, with lengths of 1.28 to 1.32 microm, 1.16 microm, and 1.05 microm, respectively. Measurements using the S1 signal confirmed the phalloidin results. Fluorescent probes may be useful to study sarcomere structure and help explain species and muscle differences in meat texture.

Side-binding proteins modulate actin filament dynamics

PubMed Central

Crevenna, Alvaro H; Arciniega, Marcelino; Dupont, Aurélie; Mizuno, Naoko; Kowalska, Kaja; Lange, Oliver F; Wedlich-Söldner, Roland; Lamb, Don C

2015-01-01

Actin filament dynamics govern many key physiological processes from cell motility to tissue morphogenesis. A central feature of actin dynamics is the capacity of filaments to polymerize and depolymerize at their ends in response to cellular conditions. It is currently thought that filament kinetics can be described by a single rate constant for each end. In this study, using direct visualization of single actin filament elongation, we show that actin polymerization kinetics at both filament ends are strongly influenced by the binding of proteins to the lateral filament surface. We also show that the pointed-end has a non-elongating state that dominates the observed filament kinetic asymmetry. Estimates of flexibility as well as effects on fragmentation and growth suggest that the observed kinetic diversity arises from structural alteration. Tuning elongation kinetics by exploiting the malleability of the filament structure may be a ubiquitous mechanism to generate a rich variety of cellular actin dynamics. DOI: http://dx.doi.org/10.7554/eLife.04599.001 PMID:25706231

Role of Intermediate Filaments in Vesicular Traffic.

PubMed

Margiotta, Azzurra; Bucci, Cecilia

2016-04-25

Intermediate filaments are an important component of the cellular cytoskeleton. The first established role attributed to intermediate filaments was the mechanical support to cells. However, it is now clear that intermediate filaments have many different roles affecting a variety of other biological functions, such as the organization of microtubules and microfilaments, the regulation of nuclear structure and activity, the control of cell cycle and the regulation of signal transduction pathways. Furthermore, a number of intermediate filament proteins have been involved in the acquisition of tumorigenic properties. Over the last years, a strong involvement of intermediate filament proteins in the regulation of several aspects of intracellular trafficking has strongly emerged. Here, we review the functions of intermediate filaments proteins focusing mainly on the recent knowledge gained from the discovery that intermediate filaments associate with key proteins of the vesicular membrane transport machinery. In particular, we analyze the current understanding of the contribution of intermediate filaments to the endocytic pathway.

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.

Solid friction between soft filaments

NASA Astrophysics Data System (ADS)

Ward, Andrew; Hilitski, Feodor; Schwenger, Walter; Welch, David; Lau, A. W. C.; Vitelli, Vincenzo; Mahadevan, L.; Dogic, Zvonimir

2015-06-01

Any macroscopic deformation of a filamentous bundle is necessarily accompanied by local sliding and/or stretching of the constituent filaments. Yet the nature of the sliding friction between two aligned filaments interacting through multiple contacts remains largely unexplored. Here, by directly measuring the sliding forces between two bundled F-actin filaments, we show that these frictional forces are unexpectedly large, scale logarithmically with sliding velocity as in solid-like friction, and exhibit complex dependence on the filaments’ overlap length. We also show that a reduction of the frictional force by orders of magnitude, associated with a transition from solid-like friction to Stokes’s drag, can be induced by coating F-actin with polymeric brushes. Furthermore, we observe similar transitions in filamentous microtubules and bacterial flagella. Our findings demonstrate how altering a filament’s elasticity, structure and interactions can be used to engineer interfilament friction and thus tune the properties of fibrous composite materials.

Solid friction between soft filaments

DOE PAGES

Ward, Andrew; Hilitski, Feodor; Schwenger, Walter; ...

2015-03-02

Any macroscopic deformation of a filamentous bundle is necessarily accompanied by local sliding and/or stretching of the constituent filaments. Yet the nature of the sliding friction between two aligned filaments interacting through multiple contacts remains largely unexplored. Here, by directly measuring the sliding forces between two bundled F-actin filaments, we show that these frictional forces are unexpectedly large, scale logarithmically with sliding velocity as in solid-like friction, and exhibit complex dependence on the filaments’ overlap length. We also show that a reduction of the frictional force by orders of magnitude, associated with a transition from solid-like friction to Stokes’s drag,more » can be induced by coating F-actin with polymeric brushes. Furthermore, we observe similar transitions in filamentous microtubules and bacterial flagella. In conclusion, our findings demonstrate how altering a filament’s elasticity, structure and interactions can be used to engineer interfilament friction and thus tune the properties of fibrous composite materials.« less

Functional characterization of contractile vacuole isolated from Amoeba proteus.

PubMed

Nishihara, Eri; Shimmen, Teruo; Sonobe, Seiji

2004-12-01

Contractile vacuoles (CVs) released from cells of Amoeba proteus were used to analyze its function in vitro. When CV was transferred to a hypertonic medium, its volume decreased within 10 sec. When it was subsequently returned to its original medium, it quickly started swelling. However, it ruptured before recovering its initial volume. These results suggested that the CV membrane is semi-permeable and that the fluid is collected by the osmotic gradient in vivo. The water permeability of membrane of isolated CV was calculated from the rate of osmotic volume change to be 0.94 microm/sec . OsM. This high value suggested that CV membrane is equipped with water channel. CV contracted (or burst) quickly upon addition of 1 mM ATP. Contraction was induced by ATP, but not by other nucleotides, GTP, ITP, ADP, or the analogues of ATP, AMP-PNP and ATPgammaS. It was suggested that the contraction of isolated CV was caused by increase in the tension of its membrane by ATP.

Recent observations of the formation of filaments

NASA Technical Reports Server (NTRS)

Martin, Sara F.

1986-01-01

Two examples of the formation of small filaments in H alpha are described and illustrated. In both cases, the formation is seen to be the spontaneous appearance of strands of absorbing mass that evolve from no previous structure. The initial development of the filaments appears to consist of the accumulation of these absorptive strands along approximately parallel paths in a channel between large-scale, opposite polarity magnetic fields on either side of the filaments. The strands exhibit continuous changes in shape and degree of absorption which can be due to successive condensations resulting in new strands, mass motions within the strands, and outflow of the mass from the strands. For at least several hours before the formation of both filaments, small-scale fragments of opposite polarity, line-of-sight magnetic flux adjacent to or immediately below the filaments, and at the ends of the filaments, were cancelling. This type of magnetic flux disappearance continued during the development of the filaments and is commonly observed in association with established filaments. Cancellation is interpreted as an important evolutionary change in the magnetic field that can lead to configurations suitable for the formation of filaments.

Detecting effects of filaments on galaxy properties in the Sloan Digital Sky Survey III

NASA Astrophysics Data System (ADS)

Chen, Yen-Chi; Ho, Shirley; Mandelbaum, Rachel; Bahcall, Neta A.; Brownstein, Joel R.; Freeman, Peter E.; Genovese, Christopher R.; Schneider, Donald P.; Wasserman, Larry

2017-04-01

We study the effects of filaments on galaxy properties in the Sloan Digital Sky Survey (SDSS) Data Release 12 using filaments from the 'Cosmic Web Reconstruction' catalogue, a publicly available filament catalogue for SDSS. Since filaments are tracers of medium- to high-density regions, we expect that galaxy properties associated with the environment are dependent on the distance to the nearest filament. Our analysis demonstrates that a red galaxy or a high-mass galaxy tends to reside closer to filaments than a blue or low-mass galaxy. After adjusting the effect from stellar mass, on average, early-forming galaxies or large galaxies have a shorter distance to filaments than late-forming galaxies or small galaxies. For the main galaxy sample, all signals are very significant (>6σ). For the LOWZ and CMASS sample, the stellar mass and size are significant (>2σ). The filament effects we observe persist until z = 0.7 (the edge of the CMASS sample). Comparing our results to those using the galaxy distances from redMaPPer galaxy clusters as a reference, we find a similar result between filaments and clusters. Moreover, we find that the effect of clusters on the stellar mass of nearby galaxies depends on the galaxy's filamentary environment. Our findings illustrate the strong correlation of galaxy properties with proximity to density ridges, strongly supporting the claim that density ridges are good tracers of filaments.

ERUPTING FILAMENTS WITH LARGE ENCLOSING FLUX TUBES AS SOURCES OF HIGH-MASS THREE-PART CMEs, AND ERUPTING FILAMENTS IN THE ABSENCE OF ENCLOSING FLUX TUBES AS SOURCES OF LOW-MASS UNSTRUCTURED CMEs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hutton, Joe; Morgan, Huw, E-mail: joh9@aber.ac.uk

2015-11-01

The 3-part appearance of many coronal mass ejections (CMEs) arising from erupting filaments emerges from a large magnetic flux tube structure, consistent with the form of the erupting filament system. Other CMEs arising from erupting filaments lack a clear 3-part structure and reasons for this have not been researched in detail. This paper aims to further establish the link between CME structure and the structure of the erupting filament system and to investigate whether CMEs which lack a 3-part structure have different eruption characteristics. A survey is made of 221 near-limb filament eruptions observed from 2013 May 03 to 2014more » June 30 by Extreme UltraViolet (EUV) imagers and coronagraphs. Ninety-two filament eruptions are associated with 3-part structured CMEs, 41 eruptions are associated with unstructured CMEs. The remaining 88 are categorized as failed eruptions. For 34% of the 3-part CMEs, processing applied to EUV images reveals the erupting front edge is a pre-existing loop structure surrounding the filament, which subsequently erupts with the filament to form the leading bright front edge of the CME. This connection is confirmed by a flux-rope density model. Furthermore, the unstructured CMEs have a narrower distribution of mass compared to structured CMEs, with total mass comparable to the mass of 3-part CME cores. This study supports the interpretation of 3-part CME leading fronts as the outer boundaries of a large pre-existing flux tube. Unstructured (non 3-part) CMEs are a different family to structured CMEs, arising from the eruption of filaments which are compact flux tubes in the absence of a large system of enclosing closed field.« less

Evaluation of humoral and cellular immune responses against HSV-1 using genetic immunization by filamentous phage particles: a comparative approach to conventional DNA vaccine.

PubMed

Hashemi, Hamidreza; Bamdad, Taravat; Jamali, Abbas; Pouyanfard, Somayeh; Mohammadi, Masoumeh Gorgian

2010-02-01

Phage display is based on expressing peptides as a fusion to one of the phage coat proteins. To date, many vaccine researches have been conducted to display immunogenic peptides or mimotopes of various pathogens and tumors on the surface of filamentous bacteriophages. In recent years as a new approach to application of phages, recombinant bacteriophage lambda particles were used as DNA delivery vehicles to mammalian cells. In this study, recombinant filamentous phage whole particles were used for vaccination of mice. BALB/c mice were inoculated with filamentous phage particles containing expression cassette of Herpes simplex virus 1 (HSV-1) glycoprotein D that has essential roles in the virus attachment and entry. Both humoral and cellular immune responses were measured in the immunized mice and compared to conventional DNA vaccination. A dose-response relationship was observed in both arms of immune responses induced by recombinant filamentous phage inoculation. The results were similar to those from DNA vaccination. Filamentous phages can be considered as suitable alternative candidate vaccines because of easier and more cost-effective production and purification over plasmid DNA or bacteriophage lambda particles. 2009 Elsevier B.V. All rights reserved.

Preserved Filamentous Microbial Biosignatures in the Brick Flat Gossan, Iron Mountain, California.

PubMed

Williams, Amy J; Sumner, Dawn Y; Alpers, Charles N; Karunatillake, Suniti; Hofmann, Beda A

2015-08-01

A variety of actively precipitating mineral environments preserve morphological evidence of microbial biosignatures. One such environment with preserved microbial biosignatures is the oxidized portion of a massive sulfide deposit, or gossan, such as that at Iron Mountain, California. This gossan may serve as a mineralogical analogue to some ancient martian environments due to the presence of oxidized iron and sulfate species, and minerals that only form in acidic aqueous conditions, in both environments. Evaluating the potential biogenicity of cryptic textures in such martian gossans requires an understanding of how microbial textures form biosignatures on Earth. The iron-oxide-dominated composition and morphology of terrestrial, nonbranching filamentous microbial biosignatures may be distinctive of the underlying formation and preservation processes. The Iron Mountain gossan consists primarily of ferric oxide (hematite), hydrous ferric oxide (HFO, predominantly goethite), and jarosite group minerals, categorized into in situ gossan, and remobilized iron deposits. We interpret HFO filaments, found in both gossan types, as HFO-mineralized microbial filaments based in part on (1) the presence of preserved central filament lumina in smooth HFO mineral filaments that are likely molds of microbial filaments, (2) mineral filament formation in actively precipitating iron-oxide environments, (3) high degrees of mineral filament bending consistent with a flexible microbial filament template, and (4) the presence of bare microbial filaments on gossan rocks. Individual HFO filaments are below the resolution of the Mars Curiosity and Mars 2020 rover cameras, but sinuous filaments forming macroscopic matlike textures are resolvable. If present on Mars, available cameras may resolve these features identified as similar to terrestrial HFO filaments and allow subsequent evaluation for their biogenicity by synthesizing geochemical, mineralogical, and morphological analyses. Sinuous

Preserved filamentous microbial biosignatures in the Brick Flat gossan, Iron Mountain, California

USGS Publications Warehouse

Williams, Amy J.; Sumner, Dawn Y.; Alpers, Charles N.; Karunatillake, Suniti; Hofmann, Beda A

2015-01-01

A variety of actively precipitating mineral environments preserve morphological evidence of microbial biosignatures. One such environment with preserved microbial biosignatures is the oxidized portion of a massive sulfide deposit, or gossan, such as that at Iron Mountain, California. This gossan may serve as a mineralogical analogue to some ancient martian environments due to the presence of oxidized iron and sulfate species, and minerals that only form in acidic aqueous conditions, in both environments. Evaluating the potential biogenicity of cryptic textures in such martian gossans requires an understanding of how microbial textures form biosignatures on Earth. The iron-oxide-dominated composition and morphology of terrestrial, nonbranching filamentous microbial biosignatures may be distinctive of the underlying formation and preservation processes. The Iron Mountain gossan consists primarily of ferric oxide (hematite), hydrous ferric oxide (HFO, predominantly goethite), and jarosite group minerals, categorized into in situ gossan, and remobilized iron deposits. We interpret HFO filaments, found in both gossan types, as HFO-mineralized microbial filaments based in part on (1) the presence of preserved central filament lumina in smooth HFO mineral filaments that are likely molds of microbial filaments, (2) mineral filament formation in actively precipitating iron-oxide environments, (3) high degrees of mineral filament bending consistent with a flexible microbial filament template, and (4) the presence of bare microbial filaments on gossan rocks. Individual HFO filaments are below the resolution of the Mars Curiosity and Mars 2020 rover cameras, but sinuous filaments forming macroscopic matlike textures are resolvable. If present on Mars, available cameras may resolve these features identified as similar to terrestrial HFO filaments and allow subsequent evaluation for their biogenicity by synthesizing geochemical, mineralogical, and morphological analyses. Sinuous

A New Instrument for Thermal Electron Attachment at High Temperature: NF3 and CH3Cl Attachment Rate Constants up to 1100 K

DTIC Science & Technology

2009-03-01

attachment at high temperature: NF3 and CH3CI attachment rate constants up to 1100 K >• Q- O o o o 1,a> Bus Thomas M. Miller, l ’ Jeffrey F...Dotan. M. Menendez-Barreto. J. V. Seeley , J. S. Williamson, F. Dale, P. L. Mundis, R. A. Morris, J. F. Paulson, and A. A. Viggiano, Rev. Sci

Nonautophagic cytoplasmic vacuolation death induction in human PC-3M prostate cancer by curcumin through reactive oxygen species -mediated endoplasmic reticulum stress

PubMed Central

Lee, Wei-Jiunn; Chien, Ming-Hsien; Chow, Jyh-Ming; Chang, Junn-Liang; Wen, Yu-Ching; Lin, Yung-Wei; Cheng, Chao-Wen; Lai, Gi-Ming; Hsiao, Michael; Lee, Liang-Ming

2015-01-01

The antiapoptotic and antiautophagic abilities of cancer cells constitute a major challenge for anticancer drug treatment. Strategies for triggering nonapoptotic or nonautophagic cell death may improve therapeutic efficacy against cancer. Curcumin has been reported to exhibit cancer chemopreventive properties. Herein, we report that curcumin induced apoptosis in LNCaP, DU145, and PC-3 cells but triggered extensive cytoplasmic vacuolation in PC-3M cells. Electron microscopic images showed that the vacuoles lacked intracellular organelles and were derived from the endoplasmic reticulum (ER). Moreover, curcumin-induced vacuolation was not reversed by an apoptosis- or autophagy-related inhibitor, suggesting that vacuolation-mediated cell death differs from classical apoptotic and autophagic cell death. Mechanistic investigations revealed that curcumin treatment upregulated the ER stress markers CHOP and Bip/GRP78 and the autophagic marker LC3-II. In addition, curcumin induced ER stress by triggering ROS generation, which was supported by the finding that treating cells with the antioxidant NAC alleviated curcumin-mediated ER stress and vacuolation-mediated death. An in vivo PC-3M orthotopic prostate cancer model revealed that curcumin reduced tumor growth by inducing ROS production followed by vacuolation-mediated cell death. Overall, our results indicated that curcumin acts as an inducer of ROS production, which leads to nonapoptotic and nonautophagic cell death via increased ER stress. PMID:26013662

Solar filament material oscillations and drainage before eruption

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bi, Yi; Jiang, Yunchun; Yang, Jiayan

Both large-amplitude longitudinal (LAL) oscillations and material drainage in a solar filament are associated with the flow of material along the filament axis, often followed by an eruption. However, the relationship between these two motions and a subsequent eruption event is poorly understood. We analyze a filament eruption using EUV imaging data captured by the Atmospheric Imaging Array on board the Solar Dynamics Observatory and the Hα images from the Global Oscillation Network Group. Hours before the eruption, the filament was activated, with one of its legs undergoing a slow rising motion. The asymmetric activation inclined the filament relative tomore » the solar surface. After the active phase, LAL oscillations were observed in the inclined filament. The oscillation period increased slightly over time, which may suggest that the magnetic fields supporting the filament evolve to be flatter during the slow rising phase. After the oscillations, a significant amount of filament material was drained toward one filament endpoint, followed immediately by the violent eruption of the filament. The material drainage may further support the change in magnetic topology prior to the eruption. Moreover, we suggest that the filament material drainage could play a role in the transition from a slow to a fast rise of the erupting filament.« less

LegC3, an Effector Protein from Legionella pneumophila, Inhibits Homotypic Yeast Vacuole Fusion In Vivo and In Vitro

PubMed Central

Bennett, Terry L.; Kraft, Shannon M.; Reaves, Barbara J.; Mima, Joji; O’Brien, Kevin M.; Starai, Vincent J.

2013-01-01

During infection, the intracellular pathogenic bacterium Legionella pneumophila causes an extensive remodeling of host membrane trafficking pathways, both in the construction of a replication-competent vacuole comprised of ER-derived vesicles and plasma membrane components, and in the inhibition of normal phagosome:endosome/lysosome fusion pathways. Here, we identify the LegC3 secreted effector protein from L. pneumophila as able to inhibit a SNARE- and Rab GTPase-dependent membrane fusion pathway in vitro, the homotypic fusion of yeast vacuoles (lysosomes). This vacuole fusion inhibition appeared to be specific, as similar secreted coiled-coiled domain containing proteins from L. pneumophila, LegC7/YlfA and LegC2/YlfB, did not inhibit vacuole fusion. The LegC3-mediated fusion inhibition was reversible by a yeast cytosolic extract, as well as by a purified soluble SNARE, Vam7p. LegC3 blocked the formation of trans-SNARE complexes during vacuole fusion, although we did not detect a direct interaction of LegC3 with the vacuolar SNARE protein complexes required for fusion. Additionally, LegC3 was incapable of inhibiting a defined synthetic model of vacuolar SNARE-driven membrane fusion, further suggesting that LegC3 does not directly inhibit the activity of vacuolar SNAREs, HOPS complex, or Sec17p/18p during membrane fusion. LegC3 is likely utilized by Legionella to modulate eukaryotic membrane fusion events during pathogenesis. PMID:23437241

Filament instability under constant loads

NASA Astrophysics Data System (ADS)

Monastra, A. G.; Carusela, M. F.; D’Angelo, M. V.; Bruno, L.

2018-04-01

Buckling of semi-flexible filaments appears in different systems and scales. Some examples are: fibers in geophysical applications, microtubules in the cytoplasm of eukaryotic cells and deformation of polymers freely suspended in a flow. In these examples, instabilities arise when a system’s parameter exceeds a critical value, being the Euler force the most known. However, the complete time evolution and wavelength of buckling processes are not fully understood. In this work we solve analytically the time evolution of a filament under a constant compressive force in the small amplitude approximation. This gives an insight into the variable force scenario in terms of normal modes. The evolution is highly sensitive to the initial configuration and to the magnitude of the compressive load. This model can be a suitable approach to many different real situations.

ERUPTION OF A SOLAR FILAMENT CONSISTING OF TWO THREADS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bi Yi; Jiang Yunchun; Li Haidong

The trigger and driving mechanism for the eruption of a filament consisting of two dark threads was studied with unprecedented high cadence and resolution of He II 304 A observations made by the Atmospheric Imagining Assembly (AIA) on board the Solar Dynamics Observatory (SDO) and the observations made by the Solar Magnetic Activity Research Telescope and the Extreme Ultraviolet Imager (EUVI) telescope on board the Solar Terrestrial Relations Observatory Ahead (STEREO-A). The filament was located at the periphery of the active region NOAA 11228 and erupted on 2011 June 6. At the onset of the eruption, a turbulent filament threadmore » was found to be heated and to elongate in stride over a second one. After it rose slowly, most interestingly, the elongating thread was driven to contact and interact with the second one, and it then erupted with its southern leg being wrapped by a newly formed thread produced by the magnetic reconnection between fields carried by the two threads. Combining the observations from STEREO-A/EUVI and SDO/AIA 304 A images, the three-dimensional shape of the axis of the filament was obtained and it was found that only the southern leg of the eruptive filament underwent rotation. We suggest that the eruption was triggered by the reconnection of the turbulent filament thread and the surrounding magnetic field, and that it was mainly driven by the kink instability of the southern leg of the eruptive filament that possessed a more twisted field introduced by the reconnection-produced thread.« less

Prediction of Solar Eruptions Using Filament Metadata

NASA Astrophysics Data System (ADS)

Aggarwal, Ashna; Schanche, Nicole; Reeves, Katharine K.; Kempton, Dustin; Angryk, Rafal

2018-05-01

We perform a statistical analysis of erupting and non-erupting solar filaments to determine the properties related to the eruption potential. In order to perform this study, we correlate filament eruptions documented in the Heliophysics Event Knowledgebase (HEK) with HEK filaments that have been grouped together using a spatiotemporal tracking algorithm. The HEK provides metadata about each filament instance, including values for length, area, tilt, and chirality. We add additional metadata properties such as the distance from the nearest active region and the magnetic field decay index. We compare trends in the metadata from erupting and non-erupting filament tracks to discover which properties present signs of an eruption. We find that a change in filament length over time is the most important factor in discriminating between erupting and non-erupting filament tracks, with erupting tracks being more likely to have decreasing length. We attempt to find an ensemble of predictive filament metadata using a Random Forest Classifier approach, but find the probability of correctly predicting an eruption with the current metadata is only slightly better than chance.

Neospora caninum Recruits Host Cell Structures to Its Parasitophorous Vacuole and Salvages Lipids from Organelles

PubMed Central

Nolan, Sabrina J.; Luechtefeld, Thomas

2015-01-01

Toxoplasma gondii and Neospora caninum, which cause the diseases toxoplasmosis and neosporosis, respectively, are two closely related apicomplexan parasites. They have similar heteroxenous life cycles and conserved genomes and share many metabolic features. Despite these similarities, T. gondii and N. caninum differ in their transmission strategies and zoonotic potential. Comparative analyses of the two parasites are important to identify the unique biological features that underlie the basis of host preference and pathogenicity. T. gondii and N. caninum are obligate intravacuolar parasites; in contrast to T. gondii, events that occur during N. caninum infection remain largely uncharacterized. We examined the capability of N. caninum (Liverpool isolate) to interact with host organelles and scavenge nutrients in comparison to that of T. gondii (RH strain). N. caninum reorganizes the host microtubular cytoskeleton and attracts endoplasmic reticulum (ER), mitochondria, lysosomes, multivesicular bodies, and Golgi vesicles to its vacuole though with some notable differences from T. gondii. For example, the host ER gathers around the N. caninum parasitophorous vacuole (PV) but does not physically associate with the vacuolar membrane; the host Golgi apparatus surrounds the N. caninum PV but does not fragment into ministacks. N. caninum relies on plasma lipoproteins and scavenges cholesterol from NPC1-containing endocytic organelles. This parasite salvages sphingolipids from host Golgi Rab14 vesicles that it sequesters into its vacuole. Our data highlight a remarkable degree of conservation in the intracellular infection program of N. caninum and T. gondii. The minor differences between the two parasites related to the recruitment and rearrangement of host organelles around their vacuoles likely reflect divergent evolutionary paths. PMID:25750213

Srs2 prevents Rad51 filament formation by repetitive motion on DNA.

PubMed

Qiu, Yupeng; Antony, Edwin; Doganay, Sultan; Koh, Hye Ran; Lohman, Timothy M; Myong, Sua

2013-01-01

Srs2 dismantles presynaptic Rad51 filaments and prevents its re-formation as an anti-recombinase. However, the molecular mechanism by which Srs2 accomplishes these tasks remains unclear. Here we report a single-molecule fluorescence study of the dynamics of Rad51 filament formation and its disruption by Srs2. Rad51 forms filaments on single-stranded DNA by sequential binding of primarily monomers and dimers in a 5'-3' direction. One Rad51 molecule binds to three nucleotides, and six monomers are required to achieve a stable nucleation cluster. Srs2 exhibits ATP-dependent repetitive motion on single-stranded DNA and this activity prevents re-formation of the Rad51 filament. The same activity of Srs2 cannot prevent RecA filament formation, indicating its specificity for Rad51. Srs2's DNA-unwinding activity is greatly suppressed when Rad51 filaments form on duplex DNA. Taken together, our results reveal an exquisite and highly specific mechanism by which Srs2 regulates the Rad51 filament formation.

Multiple Filamentation of Laser Pulses in a Glass

NASA Astrophysics Data System (ADS)

Apeksimov, D. V.; Bukin, O. A.; Golik, S. S.; Zemlyanov, A. A.; Iglakova, A. N.; Kabanov, A. M.; Kuchinskaya, O. I.; Matvienko, G. G.; Oshlakov, V. K.; Petrov, A. V.; Sokolova, E. B.

2016-03-01

Results are presented of experiments on investigation of the spatial characteristics of multi-filamentation region of giga- and terawatt pulses of a Ti:sapphire laser in a glass. Dependences are obtained of the coordinate of the beginning of filamentation region, number of filaments, their distribution along the laser beam axis, and length of filaments on the pulse power. It is shown that with increasing radiation power, the number of filaments in the multi-filamentation region decreases, whereas the filament diameter has a quasiconstant value for all powers realized in the experiments. It is shown that as a certain power of the laser pulse with Gauss energy density distribution is reached, the filamentation region acquires the shape of a hollow cone with apex directed toward the radiation source.

Trigger of Successive Filament Eruptions Observed by SDO and STEREO

NASA Astrophysics Data System (ADS)

Dhara, Sajal Kumar; Belur, Ravindra; Kumar, Pankaj; Banyal, Ravinder Kumar; Mathew, Shibu K.; Joshi, Bhuwan

2017-10-01

Using multiwavelength observations from the Solar Dynamics Observatory (SDO) and the Solar Terrestrial Relations Observatory (STEREO), we investigate the mechanism of two successive eruptions (F1 and F2) of a filament in active region NOAA 11444 on 27 March 2012. The filament was inverse J-shaped and lay along a quasi-circular polarity inversion line (PIL). The first part of the filament erupted at ˜2{:}30 UT on 27 March 2012 (F1), the second part at around 4:20 UT on the same day (F2). A precursor or preflare brightening was observed below the filament main axis about 30 min before F1. The brightening was followed by a jet-like ejection below the filament, which triggered its eruption. Before the eruption of F2, the filament seemed to be trapped within the overlying arcade loops for almost 1.5 h before it successfully erupted. Interestingly, we observe simultaneously contraction (˜12 km s^{-1}) and expansion (˜20 km s^{-1}) of arcade loops in the active region before F2. Magnetograms obtained with the Helioseismic and Magnetic Imager (HMI) show converging motion of the opposite polarities, which result in flux cancellation near the PIL. We suggest that flux cancellation at the PIL resulted in a jet-like ejection below the filament main axis, which triggered F1, similar to the tether-cutting process. F2 was triggered by removal of the overlying arcade loops via reconnection. Both filament eruptions produced high-speed (˜1000 km s^{-1}) coronal mass ejections.

Electron attachment to C{sub 2} fluorocarbon radicals at high temperature

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shuman, Nicholas S.; Miller, Thomas M.; Viggiano, Albert A., E-mail: afrl.rvborgmailbox@kirtland.af.mil

Thermal electron attachment to the radical species C{sub 2}F{sub 3} and C{sub 2}F{sub 5} has been studied over the temperature range 300–890 K using the Variable Electron and Neutral Density Attachment Mass Spectrometry technique. Both radicals exclusively undergo dissociative attachment to yield F{sup −}. The rate constant for C{sub 2}F{sub 5} shows little dependence over the temperature range, remaining ∼4 × 10{sup −9} cm{sup 3} s{sup −1}. The rate constant for C{sub 2}F{sub 3} attachment rises steeply with temperature from 3 × 10{sup −11} cm{sup 3} s{sup −1} at 300 K to 1 × 10{sup −9} cm{sup 3} s{sup −1} at 890 K.more » The behaviors of both species at high temperature are in agreement with extrapolations previously made from data below 600 K using a recently developed kinetic modeling approach. Measurements were also made on C{sub 2}F{sub 3}Br and C{sub 2}F{sub 5}Br (used in this work as precursors to the radicals) over the same temperature range, and, for C{sub 2}F{sub 5}Br as a function of electron temperature. The attachment rate constants to both species rise with temperature following Arrhenius behavior. The attachment rate constant to C{sub 2}F{sub 5}Br falls with increasing electron temperature, in agreement with the kinetic modeling. The current data fall in line with past predictions of the kinetic modeling approach, again showing the utility of this simplified approach.« less

Measuring Filament Orientation: A New Quantitative, Local Approach

DOE Office of Scientific and Technical Information (OSTI.GOV)

Green, C.-E.; Cunningham, M. R.; Jones, P. A.

The relative orientation between filamentary structures in molecular clouds and the ambient magnetic field provides insight into filament formation and stability. To calculate the relative orientation, a measurement of filament orientation is first required. We propose a new method to calculate the orientation of the one-pixel-wide filament skeleton that is output by filament identification algorithms such as filfinder. We derive the local filament orientation from the direction of the intensity gradient in the skeleton image using the Sobel filter and a few simple post-processing steps. We call this the “Sobel-gradient method.” The resulting filament orientation map can be compared quantitativelymore » on a local scale with the magnetic field orientation map to then find the relative orientation of the filament with respect to the magnetic field at each point along the filament. It can also be used for constructing radial profiles for filament width fitting. The proposed method facilitates automation in analyses of filament skeletons, which is imperative in this era of “big data.”.« less

Measuring Filament Orientation: A New Quantitative, Local Approach

NASA Astrophysics Data System (ADS)

Green, C.-E.; Dawson, J. R.; Cunningham, M. R.; Jones, P. A.; Novak, G.; Fissel, L. M.

2017-09-01

The relative orientation between filamentary structures in molecular clouds and the ambient magnetic field provides insight into filament formation and stability. To calculate the relative orientation, a measurement of filament orientation is first required. We propose a new method to calculate the orientation of the one-pixel-wide filament skeleton that is output by filament identification algorithms such as filfinder. We derive the local filament orientation from the direction of the intensity gradient in the skeleton image using the Sobel filter and a few simple post-processing steps. We call this the “Sobel-gradient method.” The resulting filament orientation map can be compared quantitatively on a local scale with the magnetic field orientation map to then find the relative orientation of the filament with respect to the magnetic field at each point along the filament. It can also be used for constructing radial profiles for filament width fitting. The proposed method facilitates automation in analyses of filament skeletons, which is imperative in this era of “big data.”

A symplectic integration method for elastic filaments

NASA Astrophysics Data System (ADS)

Ladd, Tony; Misra, Gaurav

2009-03-01

Elastic rods are a ubiquitous coarse-grained model of semi-flexible biopolymers such as DNA, actin, and microtubules. The Worm-Like Chain (WLC) is the standard numerical model for semi-flexible polymers, but it is only a linearized approximation to the dynamics of an elastic rod, valid for small deflections; typically the torsional motion is neglected as well. In the standard finite-difference and finite-element formulations of an elastic rod, the continuum equations of motion are discretized in space and time, but it is then difficult to ensure that the Hamiltonian structure of the exact equations is preserved. Here we discretize the Hamiltonian itself, expressed as a line integral over the contour of the filament. This discrete representation of the continuum filament can then be integrated by one of the explicit symplectic integrators frequently used in molecular dynamics. The model systematically approximates the continuum partial differential equations, but has the same level of computational complexity as molecular dynamics and is constraint free. Numerical tests show that the algorithm is much more stable than a finite-difference formulation and can be used for high aspect ratio filaments, such as actin. We present numerical results for the deterministic and stochastic motion of single filaments.

Experimental study of infrared filaments under different initial conditions

NASA Astrophysics Data System (ADS)

Mirell, Daniel Joseph

In 1964, four years after the first working laser was constructed, long skinny damage tracks and fluorescence trails were seen inside of certain transparent media that were excited by intense light pulses [1]. What was so remarkable about these features was the narrowness of the spatial profile and their long propagation length in the beam in concert with the very high intensity of the light that would be necessary to produce them. A purely linear model of light propagation through such media was insufficient to explain the results of these experiments and hence a new area of nonlinear optics, latex coined filamentation (to describe the length, slimness, and intensity of the light field), was born. Filament studies begin with a medium that has a nonlinear index of refraction, n¯2, that interacts with an intense beam of light so as to cause it to self-focus. The n¯2 of liquid and solid transparent media is much higher than the n¯ 2 of gases and therefore a much higher intensity of laser source would need to be invented to begin the study of filaments in air. With the advent of the Ti-Sapphire Kerr-lens modelocked laser [2], working in combination with the development of the chirped pulse amplifier system in the mid-1990's, light intensities sufficient to produce filaments in air was realized. Since that time much experimental and theoretical work has been done to better understand some of the additional complexities that arise specifically in the filamentation of light in air using several different wavelengths (UV to IR) and pulsewidths (femto- to pico-seconds). Many theoretical models exist each with a different emphasis on the various physical mechanisms that may produce the features experimentally observed in filaments. The experimental work has sought to give the theoretician better data on some of the properties of filaments such as the: (a) spatial and temporal structure of the beam and of the produced plasma (that arises due to the high intensity light

Leaky, Pulsating Laser-Plasma Filaments in the Nonlinear Schrödinger Equation Approximation

NASA Astrophysics Data System (ADS)

Johnston, Tudor Wyatt

1996-11-01

For studying self-focusing and filamention of electromagnetic beams in plasmas (and other media) (including laser beams in ICF plasmas) with power well over the critical value for self-focusing, considerable success has recently been achieved using the well-known nonlinear Schrodinger equation with "saturating" nonlinearities (i.e. those for which the nonlinearity is not infinite even if the field becomes indefinitely large). High-power beams with noticeable structure can indeed break up into numerous filaments, but these filaments emerge from the phase of filament creation in states rather close to the known filament equilibria. This is seen to be due to the loss of excess power from the filaments as they are forming and pulsating, so that each filament thus tends to condense to an equilibrium state. Analysis of filaments near equilibrium is thus seen to be more relevant than anticipated, but radiation from the filament(s) can often play an important role. Concepts have been borrowed from normal quantum mechanics (e.g. Bohr oscillations at the appropriate Rabi frequency) to explain the oscillation spectrum and whether significant radiation damping is present. Apparent momentum transfer during multiple filament creation seems common. Some discussion is given of the application of this kind of analysis to the radial confinement of sub-picosecond laser pulses over distances of tens of meters. (François Vidal (INRS) is the major collaborator in this work.)

Traffic to the malaria parasite food vacuole: a novel pathway involving a phosphatidylinositol 3-phosphate-binding protein.

PubMed

McIntosh, Michael T; Vaid, Ankush; Hosgood, H Dean; Vijay, Justin; Bhattacharya, Anindita; Sahani, Mayurbhai H; Baevova, Pavlina; Joiner, Keith A; Sharma, Pushkar

2007-04-13

Phosphatidylinositol 3-phosphate (PI3P) is a key ligand for recruitment of endosomal regulatory proteins in higher eukaryotes. Subsets of these endosomal proteins possess a highly selective PI3P binding zinc finger motif belonging to the FYVE domain family. We have identified a single FYVE domain-containing protein in Plasmodium falciparum which we term FCP. Expression and mutagenesis studies demonstrate that key residues are involved in specific binding to PI3P. In contrast to FYVE proteins in other organisms, endogenous FCP localizes to a lysosomal compartment, the malaria parasite food vacuole (FV), rather than to cytoplasmic endocytic organelles. Transfections of deletion mutants further indicate that FCP is essential for trophozoite and FV maturation and that it traffics to the FV via a novel constitutive cytoplasmic to vacuole targeting pathway. This newly discovered pathway excludes the secretory pathway and is directed by a C-terminal 44-amino acid peptide domain. We conclude that an FYVE protein that might be expected to participate in vesicle targeting in the parasite cytosol instead has a vital and functional role in the malaria parasite FV.

Dynamics and mechanics of motor-filament systems

NASA Astrophysics Data System (ADS)

Kruse, K.; Jülicher, F.

2006-08-01

Motivated by the cytoskeleton of eukaryotic cells, we develop a general framework for describing the large-scale dynamics of an active filament network. In the cytoskeleton, active cross-links are formed by motor proteins that are able to induce relative motion between filaments. Starting from pair-wise interactions of filaments via such active processes, our framework is based on momentum conservation and an analysis of the momentum flux. This allows us to calculate the stresses in the filament network generated by the action of motor proteins. We derive effective theories for the filament dynamics which can be related to continuum theories of active polar gels. As an example, we discuss the stability of homogenous isotropic filament distributions in two spatial dimensions.

Attachment of toxigenic Vibrio cholerae 01 to various freshwater plants and survival with a filamentous green alga, Rhizoclonium fontanum.

PubMed

Islam, M S; Drasar, B S; Bradley, D J

1989-12-01

In Bangladesh, cholera epidemics occur twice a year. V. cholerae 01 are readily isolated from the environment only during epidemics. The interepidemic reservoirs or sites of survival and multiplication of V. cholerae are still unknown. Investigations were carried out with various fresh-water plants as possible reservoirs of V. cholerae in the environment. Attachment to and acute population changes of V. cholerae on various plant surfaces was used as a screening technique to screen a particular plant species for survival studies. Five plant species, Anabaena variabilis, Rhizoclonium fontanum, Cladophora sp., Fontinalis antipyretica and Elodea canadensis were used for attachment experiments. Among these plants, R. fontanum showed the best attachment. On the basis of attachment results, survival experiments were carried out with R. fontanum. At 0.05% salinity toxigenic V. cholerae 01 survive longer in the presence of R. fontanum than in medium without algae.

Laser femtoseconde, filamentation, nuage et orage

NASA Astrophysics Data System (ADS)

Courvoisier, F.; Boutou, V.; Kasparian, J.; Salmon, E.; Méjean, G.; Yu, J.; Wolf, J.-P.

2005-06-01

Les applications telles que le contrôle de foudre grâce aux filaments autoguidés générés par un laser femtoseconde nécessitent de propager un tel filament à travers des aérosols de gouttes d'eau. Nous montrons qu'un filament survit à son interaction avec une goutte de diamètre comparable au sien (95 μ m), ainsi qu'à des nuages d'épaisseur optique 3,2, soit 5% de transmission. Cette transmission est permise par la présence d'un “bain de photons” autour du filament. Ce bain forme un réservoir contenant une part importante de l'énergie du faisceau, en équilibre avec le filament, et favorisant son alimentation.

Automatic Segmentation and Quantification of Filamentous Structures in Electron Tomography

PubMed Central

Loss, Leandro A.; Bebis, George; Chang, Hang; Auer, Manfred; Sarkar, Purbasha; Parvin, Bahram

2016-01-01

Electron tomography is a promising technology for imaging ultrastructures at nanoscale resolutions. However, image and quantitative analyses are often hindered by high levels of noise, staining heterogeneity, and material damage either as a result of the electron beam or sample preparation. We have developed and built a framework that allows for automatic segmentation and quantification of filamentous objects in 3D electron tomography. Our approach consists of three steps: (i) local enhancement of filaments by Hessian filtering; (ii) detection and completion (e.g., gap filling) of filamentous structures through tensor voting; and (iii) delineation of the filamentous networks. Our approach allows for quantification of filamentous networks in terms of their compositional and morphological features. We first validate our approach using a set of specifically designed synthetic data. We then apply our segmentation framework to tomograms of plant cell walls that have undergone different chemical treatments for polysaccharide extraction. The subsequent compositional and morphological analyses of the plant cell walls reveal their organizational characteristics and the effects of the different chemical protocols on specific polysaccharides. PMID:28090597

Automatic Segmentation and Quantification of Filamentous Structures in Electron Tomography.

PubMed

Loss, Leandro A; Bebis, George; Chang, Hang; Auer, Manfred; Sarkar, Purbasha; Parvin, Bahram

2012-10-01

Electron tomography is a promising technology for imaging ultrastructures at nanoscale resolutions. However, image and quantitative analyses are often hindered by high levels of noise, staining heterogeneity, and material damage either as a result of the electron beam or sample preparation. We have developed and built a framework that allows for automatic segmentation and quantification of filamentous objects in 3D electron tomography. Our approach consists of three steps: (i) local enhancement of filaments by Hessian filtering; (ii) detection and completion (e.g., gap filling) of filamentous structures through tensor voting; and (iii) delineation of the filamentous networks. Our approach allows for quantification of filamentous networks in terms of their compositional and morphological features. We first validate our approach using a set of specifically designed synthetic data. We then apply our segmentation framework to tomograms of plant cell walls that have undergone different chemical treatments for polysaccharide extraction. The subsequent compositional and morphological analyses of the plant cell walls reveal their organizational characteristics and the effects of the different chemical protocols on specific polysaccharides.

Impact of Submesoscale Processes on Dynamics of Phytoplankton Filaments

DTIC Science & Technology

2015-02-12

submesoscale processes on the formation of chlorophyll a filaments during late springearlier summer, and late summer time frames. We show that during...the late summer time frame, ASC leads to the development of filaments with high values of chlorophyll a concentration along the edge of the cold jet...in contrast to the earlier summer time, when the ASC mixes phytoplankton much deeper to the area below of the euphotic depth, and chlorophyll a

A Comparison Study of an Active Region Eruptive Filament and a Neighboring Non-Eruptive Filament

NASA Astrophysics Data System (ADS)

Wu, S. T.; Jiang, C.; Feng, X. S.; Hu, Q.

2014-12-01

We perform a comparison study of an eruptive filament in the core region of AR 11283 and a nearby non-eruptive filament. The coronal magnetic field supporting these two filaments is extrapolated using our data-driven CESE-MHD-NLFFF code (Jiang et al. 2013, Jiang etal. 2014), which presents two magnetic flux ropes (FRs) in the same extrapolation box. The eruptive FR contains a bald-patch separatrix surface (BPSS) spatially co-aligned very well with a pre-eruption EUV sigmoid, which is consistent with the BPSS model for the coronal sigmoids. The numerically reproduced magnetic dips of the FRs match observations of the filaments strikingly well, which supports strongly the FR-dip model for filaments. The FR that supports the AR eruptive filament is much smaller (with a length of 3 Mm) compared with the large-scale FR holding the quiescent filament (with a length of 30 Mm). But the AR eruptive FR contains most of the magnetic free energy in the extrapolation box and holds a much higher magnetic energy density than the quiescent FR, because it resides along the main polarity inversion line (PIL) around sunspots with strong magnetic shear. Both the FRs are weakly twisted and cannot trigger kink instability. The AR eruptive FR is unstable because its axis reaches above a critical height for torus instability (TI), at which the overlying closed arcades can no longer confine the FR stably. To the contrary, the quiescent FR is firmly held down by its overlying field, as its axis apex is far below the TI threshold height. (This work is partially supported by NSF AGS-1153323 and 1062050)

The JCMT Gould Belt Survey: properties of star-forming filaments in Orion A North

NASA Astrophysics Data System (ADS)

Salji, C. J.; Richer, J. S.; Buckle, J. V.; di Francesco, J.; Hatchell, J.; Hogerheijde, M.; Johnstone, D.; Kirk, H.; Ward-Thompson, D.; JCMT GBS Consortium

2015-05-01

We develop and apply a Hessian-based filament detection algorithm to submillimetre continuum observations of Orion A North. The resultant filament radial density profiles are fitted with beam-convolved line-of-sight Plummer-profiles using Markov chain Monte Carlo techniques. The posterior distribution of the radial decay parameter demonstrates that the majority of filaments exhibit p = 1.5-3, with a mode at p = 2.2, suggesting deviation from the Ostriker p = 4 isothermal, equilibrium, self-gravitating cylinder. The spatial distribution of young stellar objects relative to the high column density filaments is investigated, yielding a lower limit on the star-forming age of the integral-shaped filament ˜1.4 Myr. Additionally, inferred lifetimes of filaments are examined which suggest long-term filament accretion, varying rates of star formation, or both. Theoretical filament stability measures are determined with the aid of HARP C18O J = 3-2 observations and indicate that the majority of filaments are gravitationally subcritical, despite the presence of young protostars. The results from this investigation are consistent with the one-dimensional accretion flow filament model recently observed in numerical simulations.

A functional aquaporin co-localizes with the vacuolar proton pyrophosphatase to acidocalcisomes and the contractile vacuole complex of Trypanosoma cruzi.

PubMed

Montalvetti, Andrea; Rohloff, Peter; Docampo, Roberto

2004-09-10

We cloned an aquaporin gene from Trypanosoma cruzi (TcAQP) that encodes a protein of 231 amino acids, which is highly hydrophobic. The protein has six putative transmembrane domains and the two signature motifs asparagine-proline-alanine (NPA) which have been shown, in other aquaporins, to be involved in the formation of an aqueous channel spanning the bilayer. TcAQP was sensitive to endo H treatment, suggesting that the protein is N-glycosylated. Oocytes of Xenopus laevis expressing TcAQP swelled under hyposmotic conditions indicating water permeability, which was abolished after preincubating oocytes with very low concentrations of the AQP inhibitors HgCl(2) and AgNO(3). glycerol transport was detected. No Immunofluorescence microscopy of T. cruzi expressing GFP-TcAQP showed co-localization of TcAQP with the vacuolar proton pyrophosphatase (V-H(+)-PPase), a marker of acidocalcisomes. This localization was confirmed by Western blotting and immunofluorescence staining using polyclonal antibodies against a C-terminal peptide of TcAQP. In addition, there was a strong anterior labeling in a vacuole, close to the flagellar pocket, that was distinct from the acidocalcisomes and that was identified by immunogold electron microscopy as the contractile vacuole complex. Taking together, the presence of an aquaporin in acidocalcisomes and the contractile vacuole complex of T. cruzi, provides support for the role of these organelles in osmotic adaptations of these parasites.

Improved models of cable-to-post attachments for high-tension cable barriers.

DOT National Transportation Integrated Search

2012-05-01

Computer simulation models were developed to analyze and evaluate a new cable-to-post attachment for high-tension cable : barriers. The models replicated the performance of a keyway bolt currently used in the design of a high-tension cable : median b...

Determinants of Child Attachment in the Years Postpartum in a High-Risk Sample of Immigrant Women.

PubMed

Lecompte, Vanessa; Rousseau, Cécile

2017-10-07

Our goal was to examine maternal mental health and associated stresses in a sample of high-risk immigrant mothers, and its association with child insecure attachment in the years following childbirth. Mothers and their child (Mage = 37 months) were recruited through a Health and Social Service organization in the Parc-Extension neighborhood in Montreal, Quebec. Mothers completed the Hopkins Symptoms Checklist (HSCL-25), the Multidimensional Scale of Perceived Social Support (MPSS) and a sociodemographic questionnaire that included questions on premature delivery and birth weight. Attachment behaviors were coded out of a videotaped free play sequence using the Preschool and Early School-Age Attachment Rating Scales (PARS). Analysis revealed high levels of clinical anxiety and depression, low social support and low attachment security. Significant mean differences and associations were found between anxiety, depression, social support, preterm delivery and child attachment. These results underscore the importance of screening for anxiety and depression early in the postnatal years, in order to prevent associated consequences such as child insecure attachment. Results also highlight the importance of building positive social networks, especially with immigrant populations.

Filamentous Phage: Structure and Biology.

PubMed

Rakonjac, Jasna; Russel, Marjorie; Khanum, Sofia; Brooke, Sam J; Rajič, Marina

2017-01-01

Ff filamentous phage (fd, M13 and f1) of Escherichia coli have been the workhorse of phage display technology for the past 30 years. Dominance of Ff over other bacteriophage in display technology stems from the titres that are about 100-fold higher than any other known phage, efficacious transformation ensuring large library size and superior stability of the virion at high temperatures, detergents and pH extremes, allowing broad range of biopanning conditions in screening phage display libraries. Due to the excellent understanding of infection and assembly requirements, Ff phage have also been at the core of phage-assisted continual protein evolution strategies (PACE). This chapter will give an overview of the Ff filamentous phage structure and biology, emphasizing those properties of the Ff phage life cycle and virion that are pertinent to phage display applications.

Preserved Filamentous Microbial Biosignatures in the Brick Flat Gossan, Iron Mountain, California

PubMed Central

Sumner, Dawn Y.; Alpers, Charles N.; Karunatillake, Suniti; Hofmann, Beda A.

2015-01-01

Abstract A variety of actively precipitating mineral environments preserve morphological evidence of microbial biosignatures. One such environment with preserved microbial biosignatures is the oxidized portion of a massive sulfide deposit, or gossan, such as that at Iron Mountain, California. This gossan may serve as a mineralogical analogue to some ancient martian environments due to the presence of oxidized iron and sulfate species, and minerals that only form in acidic aqueous conditions, in both environments. Evaluating the potential biogenicity of cryptic textures in such martian gossans requires an understanding of how microbial textures form biosignatures on Earth. The iron-oxide-dominated composition and morphology of terrestrial, nonbranching filamentous microbial biosignatures may be distinctive of the underlying formation and preservation processes. The Iron Mountain gossan consists primarily of ferric oxide (hematite), hydrous ferric oxide (HFO, predominantly goethite), and jarosite group minerals, categorized into in situ gossan, and remobilized iron deposits. We interpret HFO filaments, found in both gossan types, as HFO-mineralized microbial filaments based in part on (1) the presence of preserved central filament lumina in smooth HFO mineral filaments that are likely molds of microbial filaments, (2) mineral filament formation in actively precipitating iron-oxide environments, (3) high degrees of mineral filament bending consistent with a flexible microbial filament template, and (4) the presence of bare microbial filaments on gossan rocks. Individual HFO filaments are below the resolution of the Mars Curiosity and Mars 2020 rover cameras, but sinuous filaments forming macroscopic matlike textures are resolvable. If present on Mars, available cameras may resolve these features identified as similar to terrestrial HFO filaments and allow subsequent evaluation for their biogenicity by synthesizing geochemical, mineralogical, and morphological analyses

Thermal design of composite materials high temperature attachments

NASA Technical Reports Server (NTRS)

1972-01-01

The thermal aspects of using filamentary composite materials as primary airframe structures on advanced atmospheric entry spacecraft such as the space shuttle vehicle were investigated to identify and evaluate potential design approaches for maintaining composite structures within allowable temperature limits at thermal protection system (TPS) attachments and/or penetrations. The investigation included: (1) definition of thermophysical data for composite material structures; (2) parametric characterization and identification of the influence of the aerodynamic heating and attachment design parameters on composite material temperatures; (3) conceptual design, evaluation, and detailed thermal analyses of temperature limiting design concepts; and (4) the development of experimental data for assessment of the thermal design methodologies and data used for evaluation of the temperature-limiting design concepts. Temperature suppression attachment concepts were examined for relative merit. The simple isolator was identified as the most weight-effective concept and was selected for detail design, thermal analysis, and testing. Tests were performed on TPS standoff attachments to boron/aluminum, boron/polyimide and graphite/epoxy composite structures.

Generalized minimal principle for rotor filaments.

PubMed

Dierckx, Hans; Wellner, Marcel; Bernus, Olivier; Verschelde, Henri

2015-05-01

To a reaction-diffusion medium with an inhomogeneous anisotropic diffusion tensor D, we add a fourth spatial dimension such that the determinant of the diffusion tensor is constant in four dimensions. We propose a generalized minimal principle for rotor filaments, stating that the scroll wave filament strives to minimize its surface area in the higher-dimensional space. As a consequence, stationary scroll wave filaments in the original 3D medium are geodesic curves with respect to the metric tensor G=det(D)D(-1). The theory is confirmed by numerical simulations for positive and negative filament tension and a model with a non-stationary spiral core. We conclude that filaments in cardiac tissue with positive tension preferentially reside or anchor in regions where cardiac cells are less interconnected, such as portions of the cardiac wall with a large number of cleavage planes.

New views of the Toxoplasma gondii parasitophorous vacuole as revealed by Helium Ion Microscopy (HIM).

PubMed

de Souza, Wanderley; Attias, Marcia

2015-07-01

The Helium Ion Microscope (HIM) is a new technology that uses a highly focused helium ion beam to scan and interact with the sample, which is not coated. The images have resolution and depth of field superior to field emission scanning electron microscopes. In this paper, we used HIM to study LLC-MK2 cells infected with Toxoplasma gondii. These samples were chemically fixed and, after critical point drying, were scraped with adhesive tape to expose the inner structure of the cell and parasitophorous vacuoles. We confirmed some of the previous findings made by field emission-scanning electron microscopy and showed that the surface of the parasite is rich in structures suggestive of secretion, that the nanotubules of the intravacuolar network (IVN) are not always straight, and that bifurcations are less frequent than previously thought. Fusion of the tubules with the parasite membrane or the parasitophorous vacuole membrane (PVM) was also infrequent. Tiny adhesive links were observed for the first time connecting the IVN tubules. The PVM showed openings of various sizes that even allowed the observation of endoplasmic reticulum membranes in the cytoplasm of the host cell. These findings are discussed in relation to current knowledge on the cell biology of T. gondii. Copyright © 2015 Elsevier Inc. All rights reserved.

Stochastic Severing of Actin Filaments by Actin Depolymerizing Factor/Cofilin Controls the Emergence of a Steady Dynamical Regime

PubMed Central

Roland, Jeremy; Berro, Julien; Michelot, Alphée; Blanchoin, Laurent; Martiel, Jean-Louis

2008-01-01

Actin dynamics (i.e., polymerization/depolymerization) powers a large number of cellular processes. However, a great deal remains to be learned to explain the rapid actin filament turnover observed in vivo. Here, we developed a minimal kinetic model that describes key details of actin filament dynamics in the presence of actin depolymerizing factor (ADF)/cofilin. We limited the molecular mechanism to 1), the spontaneous growth of filaments by polymerization of actin monomers, 2), the ageing of actin subunits in filaments, 3), the cooperative binding of ADF/cofilin to actin filament subunits, and 4), filament severing by ADF/cofilin. First, from numerical simulations and mathematical analysis, we found that the average filament length, 〈L〉, is controlled by the concentration of actin monomers (power law: 5/6) and ADF/cofilin (power law: −2/3). We also showed that the average subunit residence time inside the filament, 〈T〉, depends on the actin monomer (power law: −1/6) and ADF/cofilin (power law: −2/3) concentrations. In addition, filament length fluctuations are ∼20% of the average filament length. Moreover, ADF/cofilin fragmentation while modulating filament length keeps filaments in a high molar ratio of ATP- or ADP-Pi versus ADP-bound subunits. This latter property has a protective effect against a too high severing activity of ADF/cofilin. We propose that the activity of ADF/cofilin in vivo is under the control of an affinity gradient that builds up dynamically along growing actin filaments. Our analysis shows that ADF/cofilin regulation maintains actin filaments in a highly dynamical state compatible with the cytoskeleton dynamics observed in vivo. PMID:18065447

Striation and convection in penumbral filaments

NASA Astrophysics Data System (ADS)

Spruit, H. C.; Scharmer, G. B.; Löfdahl, M. G.

2010-10-01

Observations with the 1-m Swedish Solar Telescope of the flows seen in penumbral filaments are presented. Time sequences of bright filaments show overturning motions strikingly similar to those seen along the walls of small isolated structures in the active regions. The filaments show outward propagating striations with inclination angles suggesting that they are aligned with the local magnetic field. We interpret it as the equivalent of the striations seen in the walls of small isolated magnetic structures. Their origin is then a corrugation of the boundary between an overturning convective flow inside the filament and the magnetic field wrapping around it. The outward propagation is a combination of a pattern motion due to the downflow observed along the sides of bright filaments, and the Evershed flow. The observed short wavelength of the striation argues against the existence of a dynamically significant horizontal field inside the bright filaments. Its intensity contrast is explained by the same physical effect that causes the dark cores of filaments, light bridges and “canals”. In this way striation represents an important clue to the physics of penumbral structure and its relation with other magnetic structures on the solar surface. We put this in perspective with results from the recent 3-D radiative hydrodynamic simulations. 4 movies are only available in electronic form at http://www.aanda.org

Activation of the Saccharomyces cerevisiae filamentation/invasion pathway by osmotic stress in high-osmolarity glycogen pathway mutants

NASA Technical Reports Server (NTRS)

Davenport, K. D.; Williams, K. E.; Ullmann, B. D.; Gustin, M. C.; McIntire, L. V. (Principal Investigator)

1999-01-01

Mitogen-activated protein kinase (MAPK) cascades are frequently used signal transduction mechanisms in eukaryotes. Of the five MAPK cascades in Saccharomyces cerevisiae, the high-osmolarity glycerol response (HOG) pathway functions to sense and respond to hypertonic stress. We utilized a partial loss-of-function mutant in the HOG pathway, pbs2-3, in a high-copy suppressor screen to identify proteins that modulate growth on high-osmolarity media. Three high-copy suppressors of pbs2-3 osmosensitivity were identified: MSG5, CAK1, and TRX1. Msg5p is a dual-specificity phosphatase that was previously demonstrated to dephosphorylate MAPKs in yeast. Deletions of the putative MAPK targets of Msg5p revealed that kss1delta could suppress the osmosensitivity of pbs2-3. Kss1p is phosphorylated in response to hyperosmotic shock in a pbs2-3 strain, but not in a wild-type strain nor in a pbs2-3 strain overexpressing MSG5. Both TEC1 and FRE::lacZ expressions are activated in strains lacking a functional HOG pathway during osmotic stress in a filamentation/invasion-pathway-dependent manner. Additionally, the cellular projections formed by a pbs2-3 mutant on high osmolarity are absent in strains lacking KSS1 or STE7. These data suggest that the loss of filamentation/invasion pathway repression contributes to the HOG mutant phenotype.

Activation of the Saccharomyces cerevisiae filamentation/invasion pathway by osmotic stress in high-osmolarity glycogen pathway mutants.

PubMed Central

Davenport, K D; Williams, K E; Ullmann, B D; Gustin, M C

1999-01-01

Mitogen-activated protein kinase (MAPK) cascades are frequently used signal transduction mechanisms in eukaryotes. Of the five MAPK cascades in Saccharomyces cerevisiae, the high-osmolarity glycerol response (HOG) pathway functions to sense and respond to hypertonic stress. We utilized a partial loss-of-function mutant in the HOG pathway, pbs2-3, in a high-copy suppressor screen to identify proteins that modulate growth on high-osmolarity media. Three high-copy suppressors of pbs2-3 osmosensitivity were identified: MSG5, CAK1, and TRX1. Msg5p is a dual-specificity phosphatase that was previously demonstrated to dephosphorylate MAPKs in yeast. Deletions of the putative MAPK targets of Msg5p revealed that kss1delta could suppress the osmosensitivity of pbs2-3. Kss1p is phosphorylated in response to hyperosmotic shock in a pbs2-3 strain, but not in a wild-type strain nor in a pbs2-3 strain overexpressing MSG5. Both TEC1 and FRE::lacZ expressions are activated in strains lacking a functional HOG pathway during osmotic stress in a filamentation/invasion-pathway-dependent manner. Additionally, the cellular projections formed by a pbs2-3 mutant on high osmolarity are absent in strains lacking KSS1 or STE7. These data suggest that the loss of filamentation/invasion pathway repression contributes to the HOG mutant phenotype. PMID:10545444

Differential compartmentation of sucrose and gentianose in the cytosol and vacuoles of storage root protoplasts from Gentiana Lutea L.

PubMed

Keller, F; Wiemken, A

1982-12-01

The storage roots of perennial Gentiana lutea L.plants contain several sugars. The predominant carbohydrate reserve is gentianose (β-D-glucopyranosyl-(1 → 6)-α-D-glucopyranosyl-(1 ↔ 2)-β-D-fructofuranoside). Vacuoles were isolated from root protoplasts and purified through a betaine density gradient. The yield was about 75%. Gentianose and gentiobiose were localized to 100% in the vacuoles, fructose and glucose to about 80%, and sucrose to only about 50%. Taking the volumes of the vacuolar and extravacuolar (cytosolic) compartments into account it is inferred that gentianose is located exclusively in the vacuoles, whilst sucrose is much more concentrated in the cytosol where it may play a role as a cryoprotectant. The concentration of fructose and glucose appeared to be similar on both sides of the tonoplast.

Plasmodium falciparum: food vacuole localization of nitric oxide-derived species in intraerythrocytic stages of the malaria parasite

PubMed Central

Ostera, Graciela; Tokumasu, Fuyuki; Oliveira, Fabiano; Sa, Juliana; Furuya, Tetsuya; Teixeira, Clarissa; Dvorak, James

2008-01-01

Nitric oxide (NO) has diverse biological functions. Numerous studies have documented NO’s biosynthetic pathway in a wide variety of organisms. Little is known, however, about NO production in intraerythrocytic Plasmodium falciparum. Using diaminorhodamine-4-methyl acetoxymethylester (DAR-4M AM), a fluorescent indicator, we obtained direct evidence of NO and NO-derived reactive nitrogen species (RNS) production in intraerythrocytic P. falciparum parasites, as well as in isolated food vacuoles from trophozoite stage parasites. We preliminarily identified two gene sequences that might be implicated in NO synthesis in intraerythrocytic P. falciparum. We showed localization of the protein product of one of these two genes, a molecule that is structurally similar to a plant nitrate reductase, in trophozoite food vacuole membranes. We confirmed previous reports on the antiproliferative effect of NOS (nitric oxide synthase) inhibitors in P.falciparum cultures; however, we did not obtain evidence that NOS inhibitors had the ability to inhibit RNS production or that there is an active NOS in mature forms of the parasite. We concluded that a nitrate reductase activity produce NO and NO-derived RNS in or around the food vacuole in P. falciparum parasites. The food vacuole is a critical parasitic compartment involved in hemoglobin degradation, heme detoxification and a target for antimalarial drug action. Characterization of this relatively unexplored synthetic activity could provide important clues into poorly understood metabolic processes of the malaria parasite, PMID:18504040

Methods for modeling cytoskeletal and DNA filaments

NASA Astrophysics Data System (ADS)

Andrews, Steven S.

2014-02-01

This review summarizes the models that researchers use to represent the conformations and dynamics of cytoskeletal and DNA filaments. It focuses on models that address individual filaments in continuous space. Conformation models include the freely jointed, Gaussian, angle-biased chain (ABC), and wormlike chain (WLC) models, of which the first three bend at discrete joints and the last bends continuously. Predictions from the WLC model generally agree well with experiment. Dynamics models include the Rouse, Zimm, stiff rod, dynamic WLC, and reptation models, of which the first four apply to isolated filaments and the last to entangled filaments. Experiments show that the dynamic WLC and reptation models are most accurate. They also show that biological filaments typically experience strong hydrodynamic coupling and/or constrained motion. Computer simulation methods that address filament dynamics typically compute filament segment velocities from local forces using the Langevin equation and then integrate these velocities with explicit or implicit methods; the former are more versatile and the latter are more efficient. Much remains to be discovered in biological filament modeling. In particular, filament dynamics in living cells are not well understood, and current computational methods are too slow and not sufficiently versatile. Although primarily a review, this paper also presents new statistical calculations for the ABC and WLC models. Additionally, it corrects several discrepancies in the literature about bending and torsional persistence length definitions, and their relations to flexural and torsional rigidities.

Organizing Multiple Femtosecond Filaments in Air

NASA Astrophysics Data System (ADS)

Méchain, G.; Couairon, A.; Franco, M.; Prade, B.; Mysyrowicz, A.

2004-07-01

We show that it is possible to organize regular filamentation patterns in air by imposing either strong field gradients or phase distortions in the input-beam profile of an intense femtosecond laser pulse. A comparison between experiments and 3+1 dimensional numerical simulations confirms this concept and shows for the first time that a control of the transport of high intensities over long distances may be achieved by forcing this well ordered propagation regime. In this case, deterministic effects prevail in multiple femtosecond filamentation, and no transition to the optical turbulence regime is obtained [

The velocity characteristics of dusty filaments in the JCMT GBS clouds

NASA Astrophysics Data System (ADS)

Buckle, J. V.; Salji, C.; Richer, J. S.

2013-07-01

Large scale, high resolution spectral and continuum imaging maps have revealed, to an unprecedented extent, the characteristics of filamentary structure in star-forming molecular clouds, and their close association with star-forming cores. The filaments are associated with the formation of dense molecular cores where star formation occurs, and recent models highlight the important relationship between filaments and star-forming clusters. Velocity-coherent filaments have been proposed as the parent structures of star forming cores in Taurus. In Serpens, accretion flows along filaments have been proposed as the continuous source of mass for the star forming cluster. An evolutionary scenario for filaments based on velocity dispersion and column density measurements has recently been proposed, which we test with large scale molecular line and dust continuum maps. The JCMT Gould Belt Survey with SCUBA-2 and HARP provides dust continuum observations at 850 and 450 micron, and 12CO/13CO/C18O J=3-2 spectral line mapping of several nearby molecular clouds, covering large angular scales at high resolution. Velocities and linewidths of optically thin species, such as C18O which traces the warm, dense gas associated with star formation, are critical for an estimate of the virial stability of filamentary structures. The data and analyses that we present provide robust statistics over a large range of starless and protostellar evolutionary states. We present the velocity characteristics of dusty filaments in Orion, probing the physics at the boundary of filamentary structure and star formation. Using C18O, we investigate the internal structure of filaments, based on fragmentation and velocity coherence in the molecular line data. Through velocity dispersion measurements, we determine whether the filamentary structures are bound, and compare results between clouds of different star formation characteristics.

A Survery of the Correlation between Filament Chirality and Sigmoid Handedness

NASA Astrophysics Data System (ADS)

V, A.; Hazra, S.; Martin, S. F.; Martens, P. C.

2017-12-01

Sigmoid regions on the Sun are often the regions that cause Coronal Mass Ejections (CMEs). Large CMEs most often have filaments that erupt with them. This study focuses on the statistical relevance of the shape of the sigmoid and the chirality of the filament residing in these sigmoids. The study further extends to the relation between the directionality of filaments and the Earth-directed CMEs. Sigmoid data from Savcheva et al. (2014) between 2007 and 2012 and a compilation of data using the HEK Sigmoid Sniffer (Martens et al. 2012) along with Hinode XRT Soft X-ray images were used for analyzing data between 2013 and 2017. Hence this dataset consists of almost one solar cycle of data. A similar study done previously by Martens et al. (2013) analysed data for a solar cycle using an Advanced Automated Filament Detection & Characterization Code (Bernasconi, Rust & Hakim 2005). Considering that automated chirality detection is not foolproof, we present this study which uses manual determination of chirality for accuracy using high resolution chromospheric images. Mainly full disk images of soft X-ray obtained from Hinode XRT (X-Ray Telescope) have been used to find and ensure the S or Z shape of sigmoids. H-alpha images obtained from BBSO and Kanzelhohe Solar Observatory (KSO) are used in determining the chirality of filaments. The resolutions of BBSO and KSO data are 1k and 4k respectively. A comparison of the analysis of the chirality of filaments using both data will be presented. Although KSO gives a 4k resolution, it is still difficult to determine the chirality of small filaments. For this reason, high resolution images of H-alpha chromospheric filaments obtained from Helio Research and Solar Observing Optical Network (SOON) have been used for further analysis of chirality of those filaments that were undeterminable using the BBSO or KSO full disk images. The results of the comparison using the different resolutions are shown. The results of the correlation

The Stokesian hydrodynamics of flexing, stretching filaments

NASA Astrophysics Data System (ADS)

Shelley, Michael J.; Ueda, Tetsuji

2000-11-01

A central element of many fundamental problems in physics and biology lies in the interaction of a viscous fluid with slender, elastic filaments. Examples arise in the dynamics of biological fibers, the motility of microscopic organisms, and in phase transitions of liquid crystals. When considering the dynamics on the scale of a single filament, the surrounding fluid can often be assumed to be inertialess and hence governed by the Stokes’ equations. A typical simplification then is to assume a local relation, along the filament, between the force per unit length exerted by the filament upon the fluid and the velocity of the filament. While this assumption can be justified through slender-body theory as the leading-order effect, this approximation is only logarithmically separated (in aspect ratio) from the next-order contribution capturing the first effects of non-local interactions mediated by the surrounding fluid; non-local interactions become increasingly important as a filament comes within proximity to itself, or another filament. Motivated by a pattern forming system in isotropic to smectic-A phase transitions, we consider the non-local Stokesian dynamics of a growing elastica immersed in a fluid. The non-local interactions of the filament with itself are included using a modification of the slender-body theory of Keller and Rubinow. This modification is asymptotically equivalent, and removes an instability of their formulation at small, unphysical length-scales. Within this system, the filament lives on a marginal stability boundary, driven by a continual process of growth and buckling. Repeated bucklings result in filament flex, which, coupled to the non-local interactions and mediated by elastic response, leads to the development of space-filling patterns. We develop numerical methods to solve this system accurately and efficiently, even in the presence of temporal stiffness and the close self-approach of the filament. While we have ignored many of the

Omecamtiv mercabil and blebbistatin modulate cardiac contractility by perturbing the regulatory state of the myosin filament

PubMed Central

Kampourakis, Thomas; Zhang, Xuemeng; Sun, Yin‐Biao

2017-01-01

Key points Omecamtiv mecarbil and blebbistatin perturb the regulatory state of the thick filament in heart muscle.Omecamtiv mecarbil increases contractility at low levels of activation by stabilizing the ON state of the thick filament.Omecamtiv mecarbil decreases contractility at high levels of activation by disrupting the acto‐myosin ATPase cycle.Blebbistatin reduces contractility by stabilizing the thick filament OFF state and inhibiting acto‐myosin ATPase.Thick filament regulation is a promising target for novel therapeutics in heart disease. Abstract Contraction of heart muscle is triggered by a transient rise in intracellular free calcium concentration linked to a change in the structure of the actin‐containing thin filaments that allows the head or motor domains of myosin from the thick filaments to bind to them and induce filament sliding. It is becoming increasingly clear that cardiac contractility is also regulated through structural changes in the thick filaments, although the molecular mechanisms underlying thick filament regulation are still relatively poorly understood. Here we investigated those mechanisms using small molecules – omecamtiv mecarbil (OM) and blebbistatin (BS) – that bind specifically to myosin and respectively activate or inhibit contractility in demembranated cardiac muscle cells. We measured isometric force and ATP utilization at different calcium and small‐molecule concentrations in parallel with in situ structural changes determined using fluorescent probes on the myosin regulatory light chain in the thick filaments and on troponin C in the thin filaments. The results show that BS inhibits contractility and actin‐myosin ATPase by stabilizing the OFF state of the thick filament in which myosin head domains are more parallel to the filament axis. In contrast, OM stabilizes the ON state of the thick filament, but inhibits contractility at high intracellular calcium concentration by disrupting the actin‐myosin ATPase

Morphology and rheology in filamentous cultivations.

PubMed

Wucherpfennig, T; Kiep, K A; Driouch, H; Wittmann, C; Krull, R

2010-01-01

Because of their metabolic diversity, high production capacity, secretion efficiency, and capability of carrying out posttranslational modifications, filamentous fungi are widely exploited as efficient cell factories in the production of metabolites, bioactive substances, and native or heterologous proteins, respectively. There is, however, a complex relationship between the morphology of these microorganisms, transport phenomena, the viscosity of the cultivation broth, and related productivity. The morphological characteristics vary between freely dispersed mycelia and distinct pellets of aggregated biomass, every growth form having a distinct influence on broth rheology. Hence, the advantages and disadvantages for mycelial or pellet cultivation have to be balanced out carefully. Because of the still inadequate understanding of the morphogenesis of filamentous microorganisms, fungal morphology is often a bottleneck of productivity in industrial production. To obtain an optimized production process, it is of great importance to gain a better understanding of the molecular and cell biology of these microorganisms as well as the relevant approaches in biochemical engineering. In this chapter, morphology and growth of filamentous fungi are described, with special attention given to specific problems as they arise from fungal growth forms; growth and mass transfer in fungal biopellets are discussed as an example. To emphasize the importance of the flow behavior of filamentous cultivation broths, an introduction to rheology is also given, reviewing important rheological models and recent studies concerning rheological parameters. Furthermore, current knowledge on morphology and productivity in relation to the environom is outlined in the last section of this review. Copyright 2010 Elsevier Inc. All rights reserved.

An investigation of the security of caregiver attachment during middle childhood in children with high-functioning autistic disorder.

PubMed

Chandler, Felicity; Dissanayake, Cheryl

2014-07-01

Previous research has investigated caregiver attachment relationships in children with autism during early childhood, with few differences found from matched control groups. However, little is known of this relationship during middle childhood (ages 8-12 years). In this study, the aim was to establish whether there are differences in the security of attachment in children with high-functioning autism compared to typically developing children. A secondary aim was to establish whether caregivers' perceptions of their child's attachment to them accorded with the children's own reports. Twenty-one children with high-functioning autism and 17 typically developing children were administered the Kerns Security Scale and the Inventory of Parent and Peer Attachment-Revised, and caregivers completed the same questionnaires from the viewpoint of their child. There were no differences between the groups in the children's and parents' reports of attachment security. Parents' and children's reports were moderately correlated on the Kerns Security Scale but were not correlated on the Inventory of Parent and Peer Attachment-Revised. The results indicate that levels of attachment security in children with high-functioning autism are not different from those in typically developing children. © The Author(s) 2013.

Spatial distribution of filament elasticity determines the migratory behaviors of a cell

PubMed Central

Harn, Hans I-Chen; Hsu, Chao-Kai; Wang, Yang-Kao; Huang, Yi-Wei; Chiu, Wen-Tai; Lin, Hsi-Hui; Cheng, Chao-Min; Tang, Ming-Jer

2016-01-01

ABSTRACT Any cellular response leading to morphological changes is highly tuned to balance the force generated from structural reorganization, provided by actin cytoskeleton. Actin filaments serve as the backbone of intracellular force, and transduce external mechanical signal via focal adhesion complex into the cell. During migration, cells not only undergo molecular changes but also rapid mechanical modulation. Here we focus on determining, the role of spatial distribution of mechanical changes of actin filaments in epithelial, mesenchymal, fibrotic and cancer cells with non-migration, directional migration, and non-directional migration behaviors using the atomic force microscopy. We found 1) non-migratory cells only generated one type of filament elasticity, 2) cells generating spatially distributed two types of filament elasticity showed directional migration, and 3) pathologic cells that autonomously generated two types of filament elasticity without spatial distribution were actively migrating non-directionally. The demonstration of spatial regulation of filament elasticity of different cell types at the nano-scale highlights the coupling of cytoskeletal function with physical characters at the sub-cellular level, and provides new research directions for migration related disease. PMID:26919488

Live-cell imaging of rice cytological changes reveals the importance of host vacuole maintenance for biotrophic invasion by blast fungus, Magnaporthe oryzae.

PubMed

Mochizuki, Susumu; Minami, Eiichi; Nishizawa, Yoko

2015-12-01

The rice blast fungus Magnaporthe oryzae grows inside living host cells. Cytological analyses by live-cell imaging have revealed characteristics of the biotrophic invasion, particularly the extrainvasive hyphal membrane (EIHM) originating from the host plasma membrane and a host membrane-rich structure, biotrophic interfacial complex (BIC). Here, we observed rice subcellular changes associated with invasive hyphal growth using various transformants expressing specifically localized fluorescent proteins. The invasive hyphae did not penetrate across but were surrounded by the host vacuolar membrane together with EIHM even after branching. High-resolution imaging of BICs revealed that the host cytosol was accumulated at BIC with aggregated EIHM and a symplastic effector, Pwl2, in a punctate form. The vacuolar membrane did not aggregate in but closely surrounded the BIC. A good correlation was observed between the early collapse of vacuoles and damage of invasive hyphae in the first-invaded cell. Furthermore, a newly developed, long-term imaging method has revealed that the central vacuole gradually shrank until collapse, which was caused by the hyphal invasion occurring earlier in the neighboring cells than in the first-invaded cells. These data suggest that M. oryzae may suppress host vacuole collapse during early infection stages for successful infection. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Convergent-Filament Nonmechanical Pump

NASA Technical Reports Server (NTRS)

Collins, Earl R., Jr.

1989-01-01

Simple device induces small flow of liquid without help of moving parts, in presence or absence of gravity. Drops of liquid move on filaments from wide end of cone to narrow end. Gradually blend with drops on adjacent filaments to form large drops with menisci. Important use expected to be returning liquid condensate in heat pipes, and collection of samples from clouds or fog.

Fine structure of Mytella falcata (Bivalvia) gill filaments.

PubMed

de Oliveira David, José Augusto; Salaroli, Renato B; Fontanetti, Carmem S

2008-01-01

Bivalve filter feeders are sessile animals that live in constant contact with water and its pollutants. Their gill is an organ highly exposed to these conditions due to its large surface and its involvement in gas exchanges and feeding. The bivalve Mytella falcata is found in estuaries of Latin America, on the Atlantic as well as the Pacific Coast. It is commonly consumed, and sometimes is the only source of protein of low-income communities. In this study, gill filaments of M. falcata were characterized using histology, histochemistry and transmission electron microscopy for future comparative studies among animals exposed to environmental pollutants. Gill filaments may be divided into abfrontal, intermediate and frontal zones. Filaments are interconnected by ciliary discs. In the center of filaments, haemocytes circulate through a haemolymph vessel internally lined by an endothelium and supported by an acellular connective tissue rich in polysaccharides and collagen. The abfrontal zone contains cuboidal cells, while the intermediate zone consists of a simple squamous epithelium. The frontal zone is composed of five columnar cell types: one absorptive, mainly characterized by the presence of pinocytic vesicles in the apical region of the cell; one secretory, rarely observed; and three ciliated with abundant mitochondria. All cells lining the filament exhibit numerous microvilli and seem to absorb substances from the environment. PAS staining was observed in mucous cells in the frontal and abfrontal zones. Bromophenol blue allowed the distinction of haemocytes and detection of a glycoprotein secretion in the secretory cells of the frontal region. The characteristics of M. falcata gill filaments observed in this study were very similar to those of other bivalves, especially other Mytilidae, and are suitable for histopathological studies on the effect of water-soluble pollutants.

Listeriolysin O Membrane Damaging Activity Involves Arc Formation and Lineaction -- Implication for Listeria monocytogenes Escape from Phagocytic Vacuole

PubMed Central

Ruan, Yi; Rezelj, Saša; Bedina Zavec, Apolonija; Anderluh, Gregor; Scheuring, Simon

2016-01-01

Listeriolysin-O (LLO) plays a crucial role during infection by Listeria monocytogenes. It enables escape of bacteria from phagocytic vacuole, which is the basis for its spread to other cells and tissues. It is not clear how LLO acts at phagosomal membranes to allow bacterial escape. The mechanism of action of LLO remains poorly understood, probably due to unavailability of suitable experimental tools that could monitor LLO membrane disruptive activity in real time. Here, we used high-speed atomic force microscopy (HS-AFM) featuring high spatio-temporal resolution on model membranes and optical microscopy on giant unilamellar vesicles (GUVs) to investigate LLO activity. We analyze the assembly kinetics of toxin oligomers, the prepore-to-pore transition dynamics and the membrane disruption in real time. We reveal that LLO toxin efficiency and mode of action as a membrane-disrupting agent varies strongly depending on the membrane cholesterol concentration and the environmental pH. We discovered that LLO is able to form arc pores as well as damage lipid membranes as a lineactant, and this leads to large-scale membrane defects. These results altogether provide a mechanistic basis of how large-scale membrane disruption leads to release of Listeria from the phagocytic vacuole in the cellular context. PMID:27104344

Vorticity filaments in two-dimensional turbulence: creation, stability and effect

NASA Astrophysics Data System (ADS)

Kevlahan, N. K.-R.; Farge, M.

1997-09-01

Vorticity filaments are characteristic structures of two-dimensional turbulence. The formation, persistence and effect of vorticity filaments are examined using a high-resolution direct numerical simulation (DNS) of the merging of two positive Gaussian vortices pushed together by a weaker negative vortex. Many intense spiral vorticity filaments are created during this interaction and it is shown using a wavelet packet decomposition that, as has been suggested, the coherent vortex stabilizes the filaments. This result is confirmed by a linear stability analysis at the edge of the vortex and by a calculation of the straining induced by the spiral structure of the filament in the vortex core. The time-averaged energy spectra for simulations using hyper-viscosity and Newtonian viscosity have slopes of [minus sign]3 and [minus sign]4 respectively. Apart from a much higher effective Reynolds number (which accounts for the difference in energy spectra), the hyper-viscous simulation has the same dynamics as the Newtonian viscosity simulation. A wavelet packet decomposition of the hyper-viscous simulation reveals that after the merger the energy spectra of the filamentary and coherent parts of the vorticity field have slopes of [minus sign]2 and [minus sign]6 respectively. An asymptotic analysis and DNS for weak external strain shows that a circular filament at a distance R from the vortex centre always reduces the deformation of a Lamb's (Gaussian) vortex in the region r[gt-or-equal, slanted]R. In the region rfilament is intense and is in the vortex core, otherwise the filament may slightly increase the deformation. The results presented here should be useful for modelling the coherent and incoherent parts of two-dimensional turbulent flows.

High troposphere O3 filament at mid-latitude: a BORTAS campaign case study

NASA Astrophysics Data System (ADS)

Aruffo, Eleonora; Peterson, David; Di Carlo, Piero; Biancofiore, Fabio; Busilacchio, Marcella; Dari Salisburgo, Cesare; Giammaria, Franco; Bauguitte, Stephane; Lee, James; Moller, Sarah; Hopkins, James; Punjabi, Shalini; Lewis, Alistair C.; Palmer, Paul; Hyer, Edward

2016-04-01

During a flight (B625, 24 July 2011) of the BORTAS campaign (BOReal forest fires on Tropospheric oxidants over the Atlantic using Aircraft and Satellites, Nova Scotia, Canada, July-August 2011), an increase in the ozone (O3) concentrations has been observed at high altitude (about 7.5 Km a.s.l.) correlated with a significant growth of total peroxy nitrates (∑PNs), CO, NO2, NOy, black carbon (BC), isoprene and other species. We will illustrate the data analysis, the Hysplit back trajectories calculation and the analysis of the meteorological/physical conditions occurred during this case study in order to demonstrate that the O3 filament measured at high altitude over the Atlantic Ocean (between Nova Scotia and the Gulf of St. Lawrence) is a consequence of boreal biomass burning fires.

Crystal structure of reovirus attachment protein σ1 in complex with sialylated oligosaccharides.

PubMed

Reiter, Dirk M; Frierson, Johnna M; Halvorson, Elizabeth E; Kobayashi, Takeshi; Dermody, Terence S; Stehle, Thilo

2011-08-01

Many viruses attach to target cells by binding to cell-surface glycans. To gain a better understanding of strategies used by viruses to engage carbohydrate receptors, we determined the crystal structures of reovirus attachment protein σ1 in complex with α-2,3-sialyllactose, α-2,6-sialyllactose, and α-2,8-di-siallylactose. All three oligosaccharides terminate in sialic acid, which serves as a receptor for the reovirus serotype studied here. The overall structure of σ1 resembles an elongated, filamentous trimer. It contains a globular head featuring a compact β-barrel, and a fibrous extension formed by seven repeating units of a triple β-spiral that is interrupted near its midpoint by a short α-helical coiled coil. The carbohydrate-binding site is located between β-spiral repeats two and three, distal from the head. In all three complexes, the terminal sialic acid forms almost all of the contacts with σ1 in an identical manner, while the remaining components of the oligosaccharides make little or no contacts. We used this structural information to guide mutagenesis studies to identify residues in σ1 that functionally engage sialic acid by assessing hemagglutination capacity and growth in murine erythroleukemia cells, which require sialic acid binding for productive infection. Our studies using σ1 mutant viruses reveal that residues 198, 202, 203, 204, and 205 are required for functional binding to sialic acid by reovirus. These findings provide insight into mechanisms of reovirus attachment to cell-surface glycans and contribute to an understanding of carbohydrate binding by viruses. They also establish a filamentous, trimeric carbohydrate-binding module that could potentially be used to endow other trimeric proteins with carbohydrate-binding properties.

Electron Attachment to Radicals and Highly-Excited States in Laser-Irradiated CCl_2F_2*

NASA Astrophysics Data System (ADS)

Pinnaduwage, Lal; Datskos, Panos

1997-10-01

We have measured electron attachment rate constants for two species produced via ArF-excimer- laser irradiated CF_2Cl_2, i.e., the CF_2Cl radical and the highly-excited electronically-excited states of CF_2Cl_2. These measurements show that while electron attachment to the fragment radical has a rate constants about an order of magnitude higher compared to the ground states of CF_2Cl_2, electron attachment to the highly- excited states have many orders of magnitude larger rate constants. To our knowledge, only one other electron attachment measurement has been conducted on molecular fragments up to now. Implications of these measurements for plasma processing discharges will be discussed. Research supported by the National Science Foundation under contract No. ECS-9626217 with the University of Tennessee, Knoxville. The Oak Ridge National Laboratory is managed by Lockheed Martin Energy Research Corp. for the U. S. DOE under contract No. DE-AC05- 96OR22464.

Filament eruption connected to protospheric activity

NASA Technical Reports Server (NTRS)

Simon, G.; Gesztelyi, L.; Schmieder, B.; Mein, N

1986-01-01

Two cases of activation of filaments that occured in regions of intense magnetic activity was studied. The simultaneous observations from Debrecen Observatory (white light and H alpha filtergram), and from Meudon Observatory (magnetogram, MSDP dopplergram and intensity maps in H alpha) gave a complementary set of data from which can be produced evidence of the influence of the photospheric magnetic field on the destabilization process of the filaments. On June 22, 1980, the eruption of the filament is associated with the motion of pores, which are manifestations of emerging flux knots. On September 3, 1980, the twisting motions in the filament are associated to the birth of a pore in its neighborhood. These observations are discussed.

A simple high-voltage high current spark gap with subnanosecond jitter triggered by femtosecond laser filamentation

NASA Astrophysics Data System (ADS)

Arantchouk, L.; Houard, A.; Brelet, Y.; Carbonnel, J.; Larour, J.; André, Y.-B.; Mysyrowicz, A.

2013-04-01

We describe a simple, sturdy, and reliable spark gap operating with air at atmospheric pressure and able to switch currents in excess of 10 kA with sub-nanosecond jitter. The spark gap is remotely triggered by a femtosecond laser filament.

Coupling between myosin head conformation and the thick filament backbone structure.

PubMed

Hu, Zhongjun; Taylor, Dianne W; Edwards, Robert J; Taylor, Kenneth A

2017-12-01

The recent high-resolution structure of the thick filament from Lethocerus asynchronous flight muscle shows aspects of thick filament structure never before revealed that may shed some light on how striated muscles function. The phenomenon of stretch activation underlies the function of asynchronous flight muscle. It is most highly developed in flight muscle, but is also observed in other striated muscles such as cardiac muscle. Although stretch activation is likely to be complex, involving more than a single structural aspect of striated muscle, the thick filament itself, would be a prime site for regulatory function because it must bear all of the tension produced by both its associated myosin motors and any externally applied force. Here we show the first structural evidence that the arrangement of myosin heads within the interacting heads motif is coupled to the structure of the thick filament backbone. We find that a change in helical angle of 0.16° disorders the blocked head preferentially within the Lethocerus interacting heads motif. This observation suggests a mechanism for how tension affects the dynamics of the myosin heads leading to a detailed hypothesis for stretch activation and shortening deactivation, in which the blocked head preferentially binds the thin filament followed by the free head when force production occurs. Copyright © 2017 Elsevier Inc. All rights reserved.

Production, characterization, and modeling of mineral filled polypropylene filaments

NASA Astrophysics Data System (ADS)

George, Brian Robert

1999-11-01

This research produced mineral filled polypropylene filaments using a variety of fillers, characterized these filaments, and attempted to model their mechanical properties with current composite models. Also, these filaments were compared with bone to determine if they are suitable for modeling the mechanical properties of bone. Fillers used consist of wollastonite, talc, calcium carbonate, titanium dioxide, and hydroxyapatite. Fillers and polypropylene chips were combined and extruded into rods with the use of a mixer. The rods were chipped up and then formed into filaments through melt extrusion utilizing a piston extruder. Filaments with volume fractions of filler of 0.05, 0.10, 0.15, and 0.20 were produced. Additionally, some methods of trying to improve the properties of these filaments were attempted, but did not result in any significant property improvements. The fillers and filaments were visually characterized with a scanning electron microscope. Cross-sections, filament outer surfaces, fracture surfaces, and longitudinal cut open surfaces were viewed in this manner. Those filaments with anisotropic filler had some oriented filler particles, while all filaments suffered from poor adhesion between the polypropylene and the filler as well as agglomerations of filler particles. Twenty specimens of each filament were tensile tested and the average tenacity, strain, and modulus were calculated. Filaments containing talc, talc and wollastonite, titanium dioxide, or hydroxyapatite suffered from a drastic transition from ductile to brittle with the addition of 0.05 volume fraction of filler. This is evidenced by the sharp decrease in strain at this volume fraction of filler when compared to the strain of the unfilled polypropylene filament. Additionally, these same filaments suffered a sharp decrease in tenacity at the same volume fraction. These instant decreases are attributed to the agglomerations of filler in the filament. Generally, the modulus of the

Effect of an Energy Reservoir on the Atmospheric Propagation of Laser-Plasma Filaments

NASA Astrophysics Data System (ADS)

Eisenmann, Shmuel; Peñano, Joseph; Sprangle, Phillip; Zigler, Arie

2008-04-01

The ability to select and stabilize a single filament during propagation of an ultrashort, high-intensity laser pulse in air makes it possible to examine the longitudinal structure of the plasma channel left in its wake. We present the first detailed measurements and numerical 3-D simulations of the longitudinal plasma density variation in a laser-plasma filament after it passes through an iris that blocks the surrounding energy reservoir. Since no compensation is available from the surrounding background energy, filament propagation is terminated after a few centimeters. For this experiment, simulations indicate that filament propagation is terminated by plasma defocusing and ionization loss, which reduces the pulse power below the effective self-focusing power. With no blockage, a plasma filament length of over a few meters was observed.

Dynamics of contracting surfactant-covered filaments

NASA Astrophysics Data System (ADS)

Kamat, Pritish; Thete, Sumeet; Xu, Qi; Basaran, Osman

2013-11-01

When drops are produced from a nozzle, a thin liquid thread connects the primary drop that is about to form to the rest of the liquid in the nozzle. Often, the thread becomes disconnected from both the primary drop and the remnant liquid mass hanging from the nozzle and thereby gives rise to a free filament. Due to surface tension, the free filament then contracts or recoils. During recoil, the filament can either contract into a single satellite droplet or break up into several small satellites. Such satellite droplets are undesirable in applications where they can, for example, cause misting in a manufacturing environment and mar product quality in ink-jet printing. In many applications, the filaments are coated with a monolayer of surfactant. In this work, we study the dynamics of contraction of slender filaments of a Newtonian fluid that are covered with a monolayer of surfactant when the surrounding fluid is a passive gas. Taking advantage of the fact that the filaments are long and slender, we use a 1D-slender-jet approximation of the governing system of equations consisting of the Navier-Stokes system and the convection-diffusion equation for surfactant transport. We solve the 1D system of equations by a finite element based numerical method.

INTERACTION OF TWO FILAMENT CHANNELS OF DIFFERENT CHIRALITIES

DOE Office of Scientific and Technical Information (OSTI.GOV)

Joshi, Navin Chandra; Magara, Tetsuya; Moon, Yong-Jae

2016-07-10

We present observations of the interactions between the two filament channels of different chiralities and associated dynamics that occurred during 2014 April 18–20. While two flux ropes of different helicity with parallel axial magnetic fields can only undergo a bounce interaction when they are brought together, the observations at first glance show that the heated plasma is moving from one filament channel to the other. The SDO /AIA 171 Å observations and the potential-field source-surface magnetic field extrapolation reveal the presence of a fan-spine magnetic configuration over the filament channels with a null point located above them. Three different eventsmore » of filament activations, partial eruptions, and associated filament channel interactions have been observed. The activation initiated in one filament channel seems to propagate along the neighboring filament channel. We believe that the activation and partial eruption of the filaments brings the field lines of flux ropes containing them closer to the null point and triggers the magnetic reconnection between them and the fan-spine magnetic configuration. As a result, the hot plasma moves along the outer spine line toward the remote point. Utilizing the present observations, for the first time we have discussed how two different-chirality filament channels can interact and show interrelation.« less

Two-step solar filament eruptions

NASA Astrophysics Data System (ADS)

Filippov, B.

2018-04-01

Coronal mass ejections (CMEs) are closely related to eruptive filaments and usually are the continuation of the same eruptive process into the upper corona. There are failed filament eruptions when a filament decelerates and stops at some greater height in the corona. Sometimes the filament after several hours starts to rise again and develops into the successful eruption with a CME formation. We propose a simple model for the interpretation of such two-step eruptions in terms of equilibrium of a flux rope in a two-scale ambient magnetic field. The eruption is caused by a slow decrease of the holding magnetic field. The presence of two critical heights for the initiation of the flux-rope vertical instability allows the flux rope to stay after the first jump some time in a metastable equilibrium near the second critical height. If the decrease of the ambient field continues, the next eruption step follows.

METHOD OF MAKING TUNGSTEN FILAMENTS

DOEpatents

Frazer, J.W.

1962-12-18

A method of making tungsten filaments is described in which the tungsten is completely free of isotope impurities in the range of masses 234 to 245 for use in mass spectrometers. The filament comprises a tantalum core generally less than 1 mil in diameter having a coating of potassium-free tantalum-diffused tungsten molecularly bonded thereto. In the preferred process of manufacture a short, thin tantalum filament is first mounted between terminal posts mounted in insulated relation through a backing plate. The tungsten is most conveniently vapor plated onto the tantalum by a tungsten carbonyl vapor decomposition method having a critical step because of the tendency of the tantalum to volatilize at the temperature of operntion of the filament. The preferred recipe comprises volatilizing tantalum by resistance henting until the current drops by about 40%, cutting the voltage back to build up the tungsten, and then gradually building the temperature back up to balance the rate of tungsten deposition with the rate of tantalum volatilization. (AEC)

Manipulation by multiple filamentation of subpicosecond TW KrF laser beam

NASA Astrophysics Data System (ADS)

Zvorykin, V. D.; Smetanin, I. V.; Ustinovskii, N. N.; Shutov, A. V.

2018-05-01

A self-focusing of TW-level subpicosecond UV KrF laser pulses in ambient air produces a few 100 randomly distributed filaments over 100-m propagation distance. A control of multiple filamentation process by a number of methods was demonstrated in the present work envisaging applications for a HV discharge guiding, remote excitation of an atmospheric air laser, MW radiation transfer by virtual plasma waveguide, as well as filamentation suppression to improve short pulse parameters in direct amplification scheme. Under the laser beam focusing, a multitude of filaments coalesced into a superfilament with highly increased intensity and plasma conductivity. A superradiant forward lasing was obtained in the superfilament around 1.07-µm wavelength of atmospheric nitrogen. A regular 2D array of a 100 superfilaments was configured over 20-m distance by Fresnel diffraction on periodic amplitude masks. Effective Kerr defocusing and a subsequent filaments suppression over 50-m distance was demonstrated in Xe due to 2-photon resonance of laser radiation with 6p state being accompanied by a narrow-angle coherent conical emission at 828-nm wavelength.

Proper horizontal photospheric flows in a filament channel

NASA Astrophysics Data System (ADS)

Schmieder, B.; Roudier, T.; Mein, N.; Mein, P.; Malherbe, J. M.; Chandra, R.

2014-04-01

Context. An extended filament in the central part of the active region NOAA 11106 crossed the central meridian on Sept. 17, 2010 in the southern hemisphere. It has been observed in Hα with the THEMIS telescope in the Canary Islands and in 304 Å with the EUV imager (AIA) onboard the Solar Dynamic Observatory (SDO). Counterstreaming along the Hα threads and bright moving blobs (jets) along the 304 Å filament channel were observed during 10 h before the filament erupted at 17:03 UT. Aims: The aim of the paper is to understand the coupling between magnetic field and convection in filament channels and relate the horizontal photospheric motions to the activity of the filament. Methods: An analysis of the proper photospheric motions using SDO/HMI continuum images with the new version of the coherent structure tracking (CST) algorithm developed to track granules, as well as the large scale photospheric flows, was performed for three hours. Using corks, we derived the passive scalar points and produced a map of the cork distribution in the filament channel. Averaging the velocity vectors in the southern hemisphere in each latitude in steps of 3.5 arcsec, we defined a profile of the differential rotation. Results: Supergranules are clearly identified in the filament channel. Diverging flows inside the supergranules are similar in and out of the filament channel. Converging flows corresponding to the accumulation of corks are identified well around the Hα filament feet and at the edges of the EUV filament channel. At these convergence points, the horizontal photospheric velocity may reach 1 km s-1, but with a mean velocity of 0.35 km s-1. In some locations, horizontal flows crossing the channel are detected, indicating eventually large scale vorticity. Conclusions: The coupling between convection and magnetic field in the photosphere is relatively strong. The filament experienced the convection motions through its anchorage points with the photosphere, which are

Plasma Brightenings in a Failed Solar Filament Eruption

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Y.; Ding, M. D., E-mail: yingli@nju.edu.cn

Failed filament eruptions are solar eruptions that are not associated with coronal mass ejections. In a failed filament eruption, the filament materials usually show some ascending and falling motions as well as generating bright EUV emissions. Here we report a failed filament eruption (SOL2016-07-22) that occurred in a quiet-Sun region observed by the Atmospheric Imaging Assembly on board the Solar Dynamics Observatory . In this event, the filament spreads out but gets confined by the surrounding magnetic field. When interacting with the ambient magnetic field, the filament material brightens up and flows along the magnetic field lines through the coronamore » to the chromosphere. We find that some materials slide down along the lifting magnetic structure containing the filament and impact the chromosphere, and through kinetic energy dissipation, cause two ribbon-like brightenings in a wide temperature range. There is evidence suggesting that magnetic reconnection occurs between the filament magnetic structure and the surrounding magnetic fields where filament plasma is heated to coronal temperatures. In addition, thread-like brightenings show up on top of the erupting magnetic fields at low temperatures, which might be produced by an energy imbalance from a fast drop of radiative cooling due to plasma rarefaction. Thus, this single event of a failed filament eruption shows the existence of a variety of plasma brightenings that may be caused by completely different heating mechanisms.« less

Identification of type and causes of filamentous bulking under Mediterranean conditions.

PubMed

Noutsopoulos, C; Andreadakis, A; Mamais, D; Gavalakis, E

2007-01-01

A national survey to identify the most common filamentous microorganisms in Greek wastewater treatment plants, to assess the extent of filamentous bulking phenomenon and to correlate the occurrence of these bacteria to specific operating parameters, was carried out for five years. According to the conclusions of this survey filamentous bulking is a widespread phenomenon in Greek wastewater treatment plants. Almost 70% of the sludge samples examined exhibited filament indices greater than 4 during the winter period. The most common filamentous species found in a decreasing order were M. parvicella, Type 0092, GALOs, Type 0041 and N. limicola. A seasonal variation of the composition of the biomass was observed. M. parvicella is the dominant species during winter periods, while T0092 is the dominant species during summer conditions. This seasonal pattern of biomass composition was followed by a similar seasonal variation of the settling characteristics in terms of Filament Index (FI) and Sludge Volume Index (SVI) values. M. parvicella's proliferation is favored in Carrousel and oxidation ditches systems especially when primary settling stage was preceeded, whereas its growth is also highly stimulated in intermittent aeration systems, even in the absence of primary settling tanks. Finally there is evidence that Bio-P systems without primary sedimentation inhibit its growth.

Fabrication of PLA Filaments and its Printable Performance

NASA Astrophysics Data System (ADS)

Liu, Wenjie; Zhou, Jianping; Ma, Yuming; Wang, Jie; Xu, Jie

2017-12-01

Fused deposition modeling (FDM) is a typical 3D printing technology and preparation of qualified filaments is the basis. In order to prepare polylactic acid (PLA) filaments suitable for personalized FDM 3D printing, this article investigated the effect of factors such as extrusion temperature and screw speed on the diameter, surface roughness and ultimate tensile stress of the obtained PLA filaments. The optimal process parameters for fabrication of qualified filaments were determined. Further, the printable performance of the obtained PLA filaments for 3D objects was preliminarily explored.

Terahertz waves radiated from two noncollinear femtosecond plasma filaments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Du, Hai-Wei; Hoshina, Hiromichi; Otani, Chiko, E-mail: otani@riken.jp

2015-11-23

Terahertz (THz) waves radiated from two noncollinear femtosecond plasma filaments with a crossing angle of 25° are investigated. The irradiated THz waves from the crossing filaments show a small THz pulse after the main THz pulse, which was not observed in those from single-filament scheme. Since the position of the small THz pulse changes with the time-delay of two filaments, this phenomenon can be explained by a model in which the small THz pulse is from the second filament. The denser plasma in the overlap region of the filaments changes the movement of space charges in the plasma, thereby changingmore » the angular distribution of THz radiation. As a result, this schematic induces some THz wave from the second filament to propagate along the path of the THz wave from the first filament. Thus, this schematic alters the direction of the THz radiation from the filamentation, which can be used in THz wave remote sensing.« less

A cosmic web filament revealed in Lyman-α emission around a luminous high-redshift quasar.

PubMed

Cantalupo, Sebastiano; Arrigoni-Battaia, Fabrizio; Prochaska, J Xavier; Hennawi, Joseph F; Madau, Piero

2014-02-06

Simulations of structure formation in the Universe predict that galaxies are embedded in a 'cosmic web', where most baryons reside as rarefied and highly ionized gas. This material has been studied for decades in absorption against background sources, but the sparseness of these inherently one-dimensional probes preclude direct constraints on the three-dimensional morphology of the underlying web. Here we report observations of a cosmic web filament in Lyman-α emission, discovered during a survey for cosmic gas fluorescently illuminated by bright quasars at redshift z ≈ 2.3. With a linear projected size of approximately 460 physical kiloparsecs, the Lyman-α emission surrounding the radio-quiet quasar UM 287 extends well beyond the virial radius of any plausible associated dark-matter halo and therefore traces intergalactic gas. The estimated cold gas mass of the filament from the observed emission-about 10(12.0 ± 0.5)/C(1/2) solar masses, where C is the gas clumping factor-is more than ten times larger than what is typically found in cosmological simulations, suggesting that a population of intergalactic gas clumps with subkiloparsec sizes may be missing in current numerical models.

God attachment, mother attachment, and father attachment in early and middle adolescence.

PubMed

Sim, Tick Ngee; Yow, Amanda Shixian

2011-06-01

The present study examined the interplay of attachment to God, attachment to mother, and attachment to father with respect to adjustment (hope, self-esteem, depression) for 130 early and 106 middle adolescents in Singapore. Results showed that the parental attachments were generally linked (in expected directions) to adjustment. God attachment, however, had unique results. At the bivariate level, God attachment was only linked to early adolescents' self-esteem. When considered together with parental attachments (including interactions), God attachment did not emerge as the key moderator in attachment interactions and yielded some unexpected results (e.g., being positively linked to depression). These results are discussed viz-a-viz the secure base and safe haven functions that God and parental attachments may play during adolescence.

Endospore-forming filamentous bacteria symbiotic in termites: ultrastructure and growth in culture of Arthromitus

NASA Technical Reports Server (NTRS)

Margulis, L.; Olendzenski, L.; Afzelius, B. A.

1990-01-01

Many morphologically distinguishable filamentous spore-forming bacteria symbiotic in the paunch (hypertrophied hindguts) of wood-eating insects have been seen since Arthromitus was first described and named as a plant by Leidy in 1850. Previous descriptions were inadequate for acceptance of the group in modern bacteriological literature. Twenty-two distinguishable arthromitids in nine different arthropod hosts are recorded on the basis of microscopic studies. Five are named, including two whose ultrastructure are detailed: Arthromitus chasei sp. nov. that lives in the damp wood-eating termite Zootermopsis angusticollis (from the west coast of North America) and Arthromitus reticulitermitidis sp. nov. from the subterranean west coast termite Reticulitermes tibialis. A pterotermiditis from the desert termite Pterotermitidis occidentis; A. zootermopsidis, also from Z. angusticollis; and A. cristatus (Leidy, 1881) from Reticulitermes flavipes of eastern North America are also named here. Characterized by trichomes that show a morphogenetic sequence from no spores through immature spores to mature spores with spore filaments, Arthromitus symbionts can be identified as members of the genus by light microscopy and habitat. Electron microscopy reveals their remarkable complexity. They attach by spore filaments to various objects including the host gut wall; their maturation extends distally toward the termite lumen. By surface sterilization of the termite, maceration of the paunch, exposure to boiling temperatures and plating on soft acetate agar, the heat resistant nature of the spores and facultatively aerobic nature of Arthromitus sp. (from Zootermopsis) was demonstrated.

On the subduction of oxygenated surface water in submesoscale cold filaments off Peru.

NASA Astrophysics Data System (ADS)

Thomsen, Soeren; Kanzow, Torsten; Colas, Francois; Echevin, Vincent; Krahmann, Gerd

2015-04-01

The Peruvian upwelling regime is characterized by pronounced submesoscale variability including filaments and sharp density fronts. Submesoscale frontal processes can drive large vertical velocities and enhance vertical tracer fluxes in the upper ocean. The associated high temporal and spatial variability poses a large challenge to observational approaches targeting submesoscale processes. In this study the role of submesoscale processes for both the ventilation of the near-coastal oxygen minimum zone off Peru and the physical-biogeochemical coupling at these scales is investigated. For our study we use satellite based sea surface temperature measurements in combination with multiple high-resolution glider observations of temperature, salinity, oxygen and chlorophyll fluorescence carried out in January and February 2013 off Peru near 14°S during active upwelling. Additionally, high-resolution regional ocean circulation model outputs (ROMS) are analysed. At the beginning of our observations a previously upwelled, productive and highly oxygenated body of water is found within the mixed layer. Subsequently, a cold filament forms and the waters are moved offshore. After the decay of the filament and the relaxation of the upwelling front, the oxygen enriched surface water is found within the previously less oxygenated thermocline suggesting the occurrence of frontal subduction. A numerical model simulation is used to analyse the evolution of passive tracers and Lagrangian floats within several upwelling filaments, whose vertical structure and hydrographic properties agree well with the observations. The simulated temporal evolution of the tracers and floats support our interpretation that the subduction of previously upwelled water indeed occurs within cold filaments off Peru. Filaments are common features within eastern boundary upwelling systems, which all encompass large oxygen minimum zones. However, most state of-the-art large and regional scale physical

Lamp automatically switches to new filament on burnout

NASA Technical Reports Server (NTRS)

Ingle, W. B.

1966-01-01

Lamp with primary and secondary filaments has a means for automatic switching to the secondary filament at primary filament burnout. Lamp failures and resultant expenses during oscillograph printing are appreciably reduced.

A comparison study of a solar active-region eruptive filament and a neighboring non-eruptive filament

NASA Astrophysics Data System (ADS)

Jiang, Chao-Wei; Wu, Shi-Tsan; Feng, Xue-Shang; Hu, Qiang

2016-01-01

Solar active region (AR) 11283 is a very magnetically complex region and it has produced many eruptions. However, there exists a non-eruptive filament in the plage region just next to an eruptive one in the AR, which gives us an opportunity to perform a comparison analysis of these two filaments. The coronal magnetic field extrapolated using our CESE-MHD-NLFFF code reveals that two magnetic flux ropes (MFRs) exist in the same extrapolation box supporting these two filaments, respectively. Analysis of the magnetic field shows that the eruptive MFR contains a bald-patch separatrix surface (BPSS) cospatial very well with a pre-eruptive EUV sigmoid, which is consistent with the BPSS model for coronal sigmoids. The magnetic dips of the non-eruptive MFRs match Hα observation of the non-eruptive filament strikingly well, which strongly supports the MFR-dip model for filaments. Compared with the non-eruptive MFR/filament (with a length of about 200 Mm), the eruptive MFR/filament is much smaller (with a length of about 20 Mm), but it contains most of the magnetic free energy in the extrapolation box and holds a much higher free energy density than the non-eruptive one. Both the MFRs are weakly twisted and cannot trigger kink instability. The AR eruptive MFR is unstable because its axis reaches above a critical height for torus instability, at which the overlying closed arcades can no longer confine the MFR stably. On the contrary, the quiescent MFR is very firmly held by its overlying field, as its axis apex is far below the torus-instability threshold height. Overall, this comparison investigation supports that an MFR can exist prior to eruption and the ideal MHD instability can trigger an MFR eruption.

Omecamtiv mercabil and blebbistatin modulate cardiac contractility by perturbing the regulatory state of the myosin filament.

PubMed

Kampourakis, Thomas; Zhang, Xuemeng; Sun, Yin-Biao; Irving, Malcolm

2018-01-01

Omecamtiv mecarbil and blebbistatin perturb the regulatory state of the thick filament in heart muscle. Omecamtiv mecarbil increases contractility at low levels of activation by stabilizing the ON state of the thick filament. Omecamtiv mecarbil decreases contractility at high levels of activation by disrupting the acto-myosin ATPase cycle. Blebbistatin reduces contractility by stabilizing the thick filament OFF state and inhibiting acto-myosin ATPase. Thick filament regulation is a promising target for novel therapeutics in heart disease. Contraction of heart muscle is triggered by a transient rise in intracellular free calcium concentration linked to a change in the structure of the actin-containing thin filaments that allows the head or motor domains of myosin from the thick filaments to bind to them and induce filament sliding. It is becoming increasingly clear that cardiac contractility is also regulated through structural changes in the thick filaments, although the molecular mechanisms underlying thick filament regulation are still relatively poorly understood. Here we investigated those mechanisms using small molecules - omecamtiv mecarbil (OM) and blebbistatin (BS) - that bind specifically to myosin and respectively activate or inhibit contractility in demembranated cardiac muscle cells. We measured isometric force and ATP utilization at different calcium and small-molecule concentrations in parallel with in situ structural changes determined using fluorescent probes on the myosin regulatory light chain in the thick filaments and on troponin C in the thin filaments. The results show that BS inhibits contractility and actin-myosin ATPase by stabilizing the OFF state of the thick filament in which myosin head domains are more parallel to the filament axis. In contrast, OM stabilizes the ON state of the thick filament, but inhibits contractility at high intracellular calcium concentration by disrupting the actin-myosin ATPase pathway. The effects of BS and OM

Quantitative Analysis of Filament Branch Orientation in Listeria Actin Comet Tails.

PubMed

Jasnin, Marion; Crevenna, Alvaro H

2016-02-23

Several bacterial and viral pathogens hijack the host actin cytoskeleton machinery to facilitate spread and infection. In particular, Listeria uses Arp2/3-mediated actin filament nucleation at the bacterial surface to generate a branched network that will help propel the bacteria. However, the mechanism of force generation remains elusive due to the lack of high-resolution three-dimensional structural data on the spatial organization of the actin mother and daughter (i.e., branch) filaments within this network. Here, we have explored the three-dimensional structure of Listeria actin tails in Xenopus laevis egg extracts using cryo-electron tomography. We found that the architecture of Listeria actin tails is shared between those formed in cells and in cell extracts. Both contained nanoscopic bundles along the plane of the substrate, where the bacterium lies, and upright filaments (also called Z filaments), both oriented tangentially to the bacterial cell wall. Here, we were able to identify actin filament intersections, which likely correspond to branches, within the tails. A quantitative analysis of putative Arp2/3-mediated branches in the actin network showed that mother filaments lie on the plane of the substrate, whereas daughter filaments have random deviations out of this plane. Moreover, the analysis revealed that branches are randomly oriented with respect to the bacterial surface. Therefore, the actin filament network does not push directly toward the surface but rather accumulates, building up stress around the Listeria surface. Our results favor a mechanism of force generation for Listeria movement where the stress is released into propulsive motion. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

X-ray-emitting filaments in the cooling flow cluster A2029

NASA Technical Reports Server (NTRS)

Sarazin, Craig L.; O'Connell, Robert W.; Mcnamara, Brian R.

1992-01-01

High-resolution X-ray observations of the cluster A2029 are presented which confirm the presence of a cooling flow, despite the lack of optical line emission or evidence for recent star formation. The cooling rate and radius are about 370 solar mass/yr and 230 kpc, respectively. Emission from the inner cooling flow is dominated by a number of X-ray-emitting filaments. This may be the first case where such inhomogeneities are clearly resolved. The filaments are theorized to be supported in part by magnetic fields and may be connected with the filaments of very strong Faraday rotation seen in several nearly cooling flows.

Deformation of a helical filament by flow and electric or magnetic fields

NASA Astrophysics Data System (ADS)

Kim, Munju; Powers, Thomas R.

2005-02-01

Motivated by recent advances in the real-time imaging of fluorescent flagellar filaments in living bacteria [Turner, Ryu, and Berg, J. Bacteriol. 82, 2793 (2000)], we compute the deformation of a helical elastic filament due to flow and external magnetic or high-frequency electric fields. Two cases of deformation due to hydrodynamic drag are considered: the compression of a filament rotated by a stationary motor and the extension of a stationary filament due to flow along the helical axis. We use Kirchhoff rod theory for the filament, and work to linear order in the deflection. Hydrodynamic forces are described first by resistive-force theory, and then for comparison by the more accurate slender-body theory. For helices with a short pitch, the deflection in axial flow predicted by slender-body theory is significantly smaller than that computed with resistive-force theory. Therefore, our estimate of the bending stiffness of a flagellar filament is smaller than that of previous workers. In our calculation of the deformation of a polarizable helix in an external field, we show that the problem is equivalent to the classical case of a helix deformed by forces applied only at the ends.

Relation between halo spin and cosmic-web filaments at z ≃ 3

NASA Astrophysics Data System (ADS)

González, Roberto E.; Prieto, Joaquin; Padilla, Nelson; Jimenez, Raul

2017-02-01

We investigate the spin evolution of dark matter haloes and their dependence on the number of connected filaments from the cosmic web at high redshift (spin-filament relation hereafter). To this purpose, we have simulated 5000 haloes in the mass range 5 × 109 h-1 M⊙ to 5 × 1011 h-1 M⊙ at z = 3 in cosmological N-body simulations. We confirm the relation found by Prieto et al. (2015) where haloes with fewer filaments have larger spin. We also found that this relation is more significant for higher halo masses, and for haloes with a passive (no major mergers) assembly history. Another finding is that haloes with larger spin or with fewer filaments have their filaments more perpendicularly aligned with the spin vector. Our results point to a picture in which the initial spin of haloes is well described by tidal torque theory and then gets subsequently modified in a predictable way because of the topology of the cosmic web, which in turn is given by the currently favoured Lambda cold dark matter (LCDM) model. Our spin-filament relation is a prediction from LCDM that could be tested with observations.

High Ringxiety: Attachment Anxiety Predicts Experiences of Phantom Cell Phone Ringing.

PubMed

Kruger, Daniel J; Djerf, Jaikob M

2016-01-01

Mobile cell phone users have reported experiencing ringing and/or vibrations associated with incoming calls and messages, only to find that no call or message had actually registered. We believe this phenomenon can be understood as a human signal detection issue, with potentially important influences from psychological attributes. We hypothesized that individuals higher in attachment anxiety would report more frequent phantom cell phone experiences, whereas individuals higher in attachment avoidance would report less frequent experiences. If these experiences are primarily psychologically related to attributes of interpersonal relationships, associations with attachment style should be stronger than for general sensation seeking. We also predicted that certain contexts would interact with attachment style to increase or decrease the likelihood of experiencing phantom cell phone calls and messages. Attachment anxiety directly predicted the frequency of phantom ringing and notification experiences, whereas attachment avoidance and sensation seeking did not directly predict frequency. Attachment anxiety and attachment avoidance interacted with contextual factors (expectations for a call or message and concerned about an issue that one may be contacted about) in the expected directions for predicting phantom cell phone experiences.

Correlation of filament distortion and RRR degradation in drawn and rolled PIT and RRP Nb 3 Sn wires

DOE PAGES

Brown, M.; Tarantini, C.; Starch, W.; ...

2016-07-11

PIT and RRP® Nb3Sn strands are being developed for high field accelerator magnet upgrades for the high luminosity LHC. Here we report a quantitative study of the shape and position of PIT filaments and RRP® sub-elements after rolling lengths of unreacted PIT and RRP® round wires to simulate cabling deformation. In the as-drawn condition, filament shape distortion occurs preferentially in the outer ring filaments. By contrast, rolling induces non-uniform shear bands that generate greater distortion of inner ring filaments. By making a full digitization of the shapes of all filaments, we find that a critical distortion occurs for thickness reductionsmore » between 10% and 20% when filament shapes in inner filament rings heavily degrade, especially in the vicinity of the strong 45° shear bands imposed by the rolling. It is well known that maintaining diffusion barrier integrity is vital to retaining adequate RRR in the stabilizing copper needed for magnet stability. Diffusion barrier breaks occur preferentially in these distorted inner filaments and drive local Sn leakage during reaction, increasing RRR degradation.« less

Pattern formation and filamentation in low temperature, magnetized plasmas - a numerical approach

NASA Astrophysics Data System (ADS)

Menati, Mohamad; Konopka, Uwe; Thomas, Edward

2017-10-01

In low-temperature discharges under the influence of high magnetic field, pattern and filament formation in the plasma has been reported by different groups. The phenomena present themselves as bright plasma columns (filaments) oriented parallel to the magnetic field lines at high magnetic field regime. The plasma structure can filament into different shapes from single columns to spiral and bright rings when viewed from the top. In spite of the extensive experimental observations, the observed effects lack a detailed theoretical and numerical description. In an attempt to numerically explain the plasma filamentation, we present a simplified model for the plasma discharge and power deposition into the plasma. Based on the model, 2-D and 3-D codes are being developed that solve Poisson's equation along with the fluid equations to obtain a self-consistent description of the plasma. The model and preliminary results applied to the specific plasma conditions will be presented. This work was supported by the US Dept. of Energy and NSF, DE-SC0016330, PHY-1613087.

Observations of the Growth of an Active Region Filament

NASA Astrophysics Data System (ADS)

Yang, Bo

2017-04-01

We present observations of the growth of an active region filament caused by magnetic interactions among the filament and its adjacent superpenumbral filament (SF) and dark thread-like structures (T). Multistep reconnections are identified during the whole growing process. Magnetic flux convergence and cancellation occurring at the positive footpoint region of the filament is the first step reconnection, which resulted in the filament bifurcating into two sets of intertwined threads. One set anchored in situ, while the other set moved toward and interacted with the SF and part of T. This indicates the second step reconnection, which gave rise to the disappearance of the SF and the formation of a long thread-like structure that connects the far ends of the filament and T. The long thread-like structure further interacted with the T and then separated into two parts, representing the third step reconnection. Finally, another similar long thread-like structure, which intertwined with the fixed filament threads, appeared. Hαobservations show that this twisted structure is a longer sinistral filament. Based on the observed photospheric vector magnetograms, we performed a non-linear force-free field extrapolation to reconstruct the magnetic fields above the photosphere and found that the coronal magnetic field lines associated with the filament consists of two twisted flux ropes winding around each other. These results suggest that magnetic interactions among filaments and their adjacent SFs and T could lead to the growth of the filaments, and the filament is probably supported in a flux rope.

OBSERVATIONS OF THE GROWTH OF AN ACTIVE REGION FILAMENT

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yang, Bo; Jiang, Yunchun; Yang, Jiayan

We present observations of the growth of an active region filament caused by magnetic interactions among the filament and its adjacent superpenumbral filament (SF) and dark thread-like structures (T). Multistep reconnections are identified during the whole growing process. Magnetic flux convergence and cancellation occurring at the positive footpoint region of the filament is the first step reconnection, which resulted in the filament bifurcating into two sets of intertwined threads. One set anchored in situ, while the other set moved toward and interacted with the SF and part of T. This indicates the second step reconnection, which gave rise to themore » disappearance of the SF and the formation of a long thread-like structure that connects the far ends of the filament and T. The long thread-like structure further interacted with the T and then separated into two parts, representing the third step reconnection. Finally, another similar long thread-like structure, which intertwined with the fixed filament threads, appeared. H {sub α} observations show that this twisted structure is a longer sinistral filament. Based on the observed photospheric vector magnetograms, we performed a non-linear force-free field extrapolation to reconstruct the magnetic fields above the photosphere and found that the coronal magnetic field lines associated with the filament consists of two twisted flux ropes winding around each other. These results suggest that magnetic interactions among filaments and their adjacent SFs and T could lead to the growth of the filaments, and the filament is probably supported in a flux rope.« less

Deleting the DAG Kinase Dgk1 Augments Yeast Vacuole Fusion Through In-creased Ypt7 Activity and Altered Membrane Fluidity

PubMed Central

Miner, Gregory E.; Starr, Matthew L.; Hurst, Logan R.; Fratti, Rutilio A.

2017-01-01

Diacylglycerol (DAG) is a fusogenic lipid that can be produced through phospholipase C activity on phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], or through phosphatidic acid (PA) phosphatase activity. The fusion of Saccharomyces cerevisiae vacuoles requires DAG, PA and PI(4,5)P2, and the production of these lipids is thought to provide temporally specific stoichiometries that are critical for each stage of fusion. Furthermore, DAG and PA can be interconverted by the DAG kinase Dgk1 and the PA phosphatase Pah1. Previously we found that pah1Δ vacuoles were fragmented, blocked in SNARE priming and showed arrested endosomal maturation. In other pathways the effects of deleting PAH1 can be compensated for by additionally deleting DGK1, however deleting both genes did not rescue the pah1Δ vacuolar defects. Deleting DGK1 alone caused a marked increase in vacuole fusion that was attributed to elevated DAG levels. This was accompanied by a gain in resistance to the inhibitory effects of PA as well as inhibitors of Ypt7 activity. Together these data show that Dgk1 function can act as a negative regulator of vacuole fusion through the production of PA at the cost of depleting DAG and reducing Ypt7 activity. PMID:28276191

Fabrication of High-Sensitivity Skin-Attachable Temperature Sensors with Bioinspired Microstructured Adhesive.

PubMed

Oh, Ju Hyun; Hong, Soo Yeong; Park, Heun; Jin, Sang Woo; Jeong, Yu Ra; Oh, Seung Yun; Yun, Junyeong; Lee, Hanchan; Kim, Jung Wook; Ha, Jeong Sook

2018-02-28

In this study, we demonstrate the fabrication of a highly sensitive flexible temperature sensor with a bioinspired octopus-mimicking adhesive. A resistor-type temperature sensor consisting of a composite of poly(N-isopropylacrylamide) (pNIPAM)-temperature sensitive hydrogel, poly(3,4-ethylenedioxythiophene) polystyrene sulfonate, and carbon nanotubes exhibits a very high thermal sensitivity of 2.6%·°C -1 between 25 and 40 °C so that the change in skin temperature of 0.5 °C can be accurately detected. At the same time, the polydimethylsiloxane adhesive layer of octopus-mimicking rim structure coated with pNIPAM is fabricated through the formation of a single mold by utilizing undercut phenomenon in photolithography. The fabricated sensor shows stable and reproducible detection of skin temperature under repeated attachment/detachment cycles onto skin without any skin irritation for a long time. This work suggests a high potential application of our skin-attachable temperature sensor to wearable devices for medical and health-care monitoring.

New insights into roles of acidocalcisomes and contractile vacuole complex in osmoregulation in protists.

PubMed

Docampo, Roberto; Jimenez, Veronica; Lander, Noelia; Li, Zhu-Hong; Niyogi, Sayantanee

2013-01-01

While free-living protists are usually subjected to hyposmotic environments, parasitic protists are also in contact with hyperosmotic habitats. Recent work in one of these parasites, Trypanosoma cruzi, has revealed that its contractile vacuole complex, which usually collects and expels excess water as a mechanism of regulatory volume decrease after hyposmotic stress, has also a role in cell shrinking when the cells are submitted to hyperosmotic stress. Trypanosomes also have an acidic calcium store rich in polyphosphate (polyP), named the acidocalcisome, which is involved in their response to osmotic stress. Here, we review newly emerging insights on the role of acidocalcisomes and the contractile vacuole complex in the cellular response to hyposmotic and hyperosmotic stresses. We also review the current state of knowledge on the composition of these organelles and their other roles in calcium homeostasis and protein trafficking. © 2013, Elsevier Inc. All Rights Reserved.

Hot-filament chemical vapor deposition chamber and process with multiple gas inlets

DOEpatents

Deng, Xunming; Povolny, Henry S.

2004-06-29

A thin film deposition method uses a vacuum confinement cup that employs a dense hot filament and multiple gas inlets. At least one reactant gas is introduced into the confinement cup both near and spaced apart from the heated filament. An electrode inside the confinement cup is used to generate plasma for film deposition. The method is used to deposit advanced thin films (such as silicon based thin films) at a high quality and at a high deposition rate.

Brownian microhydrodynamics of active filaments.

PubMed

Laskar, Abhrajit; Adhikari, R

2015-12-21

Slender bodies capable of spontaneous motion in the absence of external actuation in an otherwise quiescent fluid are common in biological, physical and technological contexts. The interplay between the spontaneous fluid flow, Brownian motion, and the elasticity of the body presents a challenging fluid-structure interaction problem. Here, we model this problem by approximating the slender body as an elastic filament that can impose non-equilibrium velocities or stresses at the fluid-structure interface. We derive equations of motion for such an active filament by enforcing momentum conservation in the fluid-structure interaction and assuming slow viscous flow in the fluid. The fluid-structure interaction is obtained, to any desired degree of accuracy, through the solution of an integral equation. A simplified form of the equations of motion, which allows for efficient numerical solutions, is obtained by applying the Kirkwood-Riseman superposition approximation to the integral equation. We use this form of equation of motion to study dynamical steady states in free and hinged minimally active filaments. Our model provides the foundation to study collective phenomena in momentum-conserving, Brownian, active filament suspensions.

SWAYING THREADS OF A SOLAR FILAMENT

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, Y.; Engvold, O.; Langangen, Oe.

From recent high-resolution observations obtained with the Swedish 1 m Solar Telescope in La Palma, we detect swaying motions of individual filament threads in the plane of the sky. The oscillatory characters of these motions are comparable with oscillatory Doppler signals obtained from corresponding filament threads. Simultaneous recordings of motions in the line of sight and in the plane of the sky give information about the orientation of the oscillatory plane. These oscillations are interpreted in the context of the magnetohydrodynamic (MHD) theory. Kink MHD waves supported by the thread body are proposed as an explanation of the observed threadmore » oscillations. On the basis of this interpretation and by means of seismological arguments, we give an estimation of the thread Alfven speed and magnetic field strength by means of seismological arguments.« less

Filament structure, organization, and dynamics in MreB sheets.

PubMed

Popp, David; Narita, Akihiro; Maeda, Kayo; Fujisawa, Tetsuro; Ghoshdastider, Umesh; Iwasa, Mitsusada; Maéda, Yuichiro; Robinson, Robert C

2010-05-21

In vivo fluorescence microscopy studies of bacterial cells have shown that the bacterial shape-determining protein and actin homolog, MreB, forms cable-like structures that spiral around the periphery of the cell. The molecular structure of these cables has yet to be established. Here we show by electron microscopy that Thermatoga maritime MreB forms complex, several mum long multilayered sheets consisting of diagonally interwoven filaments in the presence of either ATP or GTP. This architecture, in agreement with recent rheological measurements on MreB cables, may have superior mechanical properties and could be an important feature for maintaining bacterial cell shape. MreB polymers within the sheets appear to be single-stranded helical filaments rather than the linear protofilaments found in the MreB crystal structure. Sheet assembly occurs over a wide range of pH, ionic strength, and temperature. Polymerization kinetics are consistent with a cooperative assembly mechanism requiring only two steps: monomer activation followed by elongation. Steady-state TIRF microscopy studies of MreB suggest filament treadmilling while high pressure small angle x-ray scattering measurements indicate that the stability of MreB polymers is similar to that of F-actin filaments. In the presence of ADP or GDP, long, thin cables formed in which MreB was arranged in parallel as linear protofilaments. This suggests that the bacterial cell may exploit various nucleotides to generate different filament structures within cables for specific MreB-based functions.

Presynaptic Filament Dynamics in Homologous Recombination and DNA Repair

PubMed Central

Liu, Jie; Ehmsen, Kirk T.; Heyer, Wolf-Dietrich; Morrical, Scott W.

2014-01-01

Homologous Recombination (HR) is an essential genome stability mechanism used for high-fidelity repair of DNA double-strand breaks and for the recovery of stalled or collapsed DNA replication forks. The crucial homology search and DNA strand exchange steps of HR are catalyzed by presynaptic filaments—helical filaments of a recombinase enzyme bound to single-stranded DNA. Presynaptic filaments are fundamentally dynamic structures, the assembly, catalytic turnover, and disassembly of which must be closely coordinated with other elements of the DNA recombination, repair, and replication machinery in order for genome maintenance functions to be effective. Here, we review the major dynamic elements controlling the assembly, activity, and disassembly of presynaptic filaments: some intrinsic such as recombinase ATP binding and hydrolytic activities, others extrinsic such as ssDNA-binding proteins, mediator proteins, and DNA motor proteins. We examine dynamic behavior on multiple levels, including atomic- and filament-level structural changes associated with ATP binding and hydrolysis as evidenced in crystal structures, as well as subunit binding and dissociation events driven by intrinsic and extrinsic factors. We examine the biochemical properties of recombination proteins from four model systems (T4 phage, E. coli, S. cerevisiae, and H. sapiens), demonstrating how their properties are tailored for the context-specific requirements in these diverse species. We propose that the presynaptic filament has evolved to rely on multiple external factors for increased multi-level regulation of HR processes in genomes with greater structural and sequence complexity. PMID:21599536

Spontaneous beating and synchronization of extensile active filament

NASA Astrophysics Data System (ADS)

Sarkar, Debarati; Thakur, Snigdha

2017-04-01

We simulate a semi-flexible active filament that exhibits spontaneous oscillations on clamping and show self-propulsion when left free. The activity on the filament relies on the nano-dimers distributed at regular intervals along the chain. With an emphasis on the spontaneous beating of a clamped filament, we demonstrate that the two competing forces necessary for oscillation are the elastic forces due to polymer rigidity and the active forces due to chemical activity. In addition, we also study the synchronization of two extensile filaments and the role played by non-local hydrodynamic interactions. We observe a phase lock scenario between the filaments during their synchronous motion.

Hydrodynamic interaction induced spontaneous rotation of coupled active filaments.

PubMed

Jiang, Huijun; Hou, Zhonghuai

2014-12-14

We investigate the coupled dynamics of active filaments with long range hydrodynamic interactions (HI). Remarkably, we find that filaments can rotate spontaneously under the same conditions in which a single filament alone can only move in translation. Detailed analysis reveals that the emergence of coupled rotation originates from an asymmetric flow field associated with HI which breaks the symmetry of translational motion when filaments approach. The breaking is then further stabilized by HI to form self-sustained coupled rotation. Intensive simulations show that coupled rotation forms easily when one filament tends to collide with the front-half of the other. For head-to-tail approaching, we observe another interesting HI-induced coupled motion, where filaments move together in the form of one following the other. Moreover, the radius of coupled rotation increases exponentially as the rigidity of the filament increases, which suggests that HI are also important for the alignment of rigid-rod-like filaments which has been assumed to be solely a consequence of direct collisions.

Star-forming Filament Models

DOE Office of Scientific and Technical Information (OSTI.GOV)

Myers, Philip C., E-mail: pmyers@cfa.harvard.edu

2017-03-20

New models of star-forming filamentary clouds are presented in order to quantify their properties and to predict their evolution. These 2D axisymmetric models describe filaments that have no core, one low-mass core, and one cluster-forming core. They are based on Plummer-like cylinders and spheroids that are bounded by a constant-density surface of finite extent. In contrast to 1D Plummer-like models, they have specific values of length and mass, they approximate observed column density maps, and their distributions of column density ( N -pdfs) are pole-free. Each model can estimate the star-forming potential of a core-filament system by identifying the zonemore » of gas dense enough to form low-mass stars and by counting the number of enclosed thermal Jeans masses. This analysis suggests that the Musca central filament may be near the start of its star-forming life, with enough dense gas to make its first ∼3 protostars, while the Coronet filament is near the midpoint of its star formation, with enough dense gas to add ∼8 protostars to its ∼20 known stars. In contrast, L43 appears to be near the end of its star-forming life, since it lacks enough dense gas to add any new protostars to the two young stellar objectsalready known.« less

[Induction and regulation of cellulase expression in filamentous fungi: a review].

PubMed

Zhang, Fei; Bai, Fengwu; Zhao, Xinqing

2016-11-25

Production of bioenergy and bio-based chemicals by using fermentable sugars released from low-cost renewable lignocellulosic biomass has received great attention. Efficient cellulolytic enzymes are crucial for lignocellulose bioconversion, but high cellulase production cost is limiting the bioconversion efficiency of cellulosic biomass and industrial applications of lignocellulose biorefinery. Studies on induction and regulation of cellulase in filamentous fungi will help to further develop superior fungal strains for efficient cellulase production and reduce cellulase production cost. With the advances in high-throughput sequencing and gene manipulation technology using fungal strains, an in-depth understanding of cellulase induction and regulation mechanisms of enzyme expression has been achieved. We reviewed recent progresses in the induction and regulation of cellulase expression in several model filamentous fungi, emphasizing sugar transporters, transcription factors and chromatin remodeling. Future prospects in application of artificial zinc finger proteins for cellulase induction and regulation in filamentous fungi were discussed.

Prokaryotic cytoskeletons: protein filaments organizing small cells.

PubMed

Wagstaff, James; Löwe, Jan

2018-04-01

Most, if not all, bacterial and archaeal cells contain at least one protein filament system. Although these filament systems in some cases form structures that are very similar to eukaryotic cytoskeletons, the term 'prokaryotic cytoskeletons' is used to refer to many different kinds of protein filaments. Cytoskeletons achieve their functions through polymerization of protein monomers and the resulting ability to access length scales larger than the size of the monomer. Prokaryotic cytoskeletons are involved in many fundamental aspects of prokaryotic cell biology and have important roles in cell shape determination, cell division and nonchromosomal DNA segregation. Some of the filament-forming proteins have been classified into a small number of conserved protein families, for example, the almost ubiquitous tubulin and actin superfamilies. To understand what makes filaments special and how the cytoskeletons they form enable cells to perform essential functions, the structure and function of cytoskeletal molecules and their filaments have been investigated in diverse bacteria and archaea. In this Review, we bring these data together to highlight the diverse ways that linear protein polymers can be used to organize other molecules and structures in bacteria and archaea.

A Method for 3D-Reconstruction of a Muscle Thick Filament Using the Tilt Series Images of a Single Filament Electron Tomogram

PubMed Central

Márquez, G.; Pinto, A.; Alamo, L.; Baumann, B.; Ye, F.; Winkler, H.; Taylor, K.; Padrón, R.

2014-01-01

Summary Myosin interacting-heads (MIH) motifs are visualized in 3D-reconstructions of thick filaments from striated muscle. These reconstructions are calculated by averaging methods using images from electron micrographs of grids prepared using numerous filament preparations. Here we propose an alternative method to calculate the 3D-reconstruction of a single thick filament using only a tilt series images recorded by electron tomography. Relaxed thick filaments, prepared from tarantula leg muscle homogenates, were negatively stained. Single-axis tilt series of single isolated thick filaments were obtained with the electron microscope at a low electron dose, and recorded on a CCD camera by electron tomography. An IHRSR 3D-recontruction was calculated from the tilt series images of a single thick filament. The reconstruction was enhanced by including in the search stage dual tilt image segments while only single tilt along the filament axis is usually used, as well as applying a band pass filter just before the back projection. The reconstruction from a single filament has a 40 Å resolution and clearly shows the presence of MIH motifs. In contrast, the electron tomogram 3D-reconstruction of the same thick filament –calculated without any image averaging and/or imposition of helical symmetry- only reveals MIH motifs infrequently. This is –to our knowledge- the first application of the IHRSR method to calculate a 3D reconstruction from tilt series images. This single filament IHRSR reconstruction method (SF-IHRSR) should provide a new tool to assess structural differences between well-ordered thick (or thin) filaments in a grid by recording separately their electron tomograms. PMID:24727133

A method for 3D-reconstruction of a muscle thick filament using the tilt series images of a single filament electron tomogram.

PubMed

Márquez, G; Pinto, A; Alamo, L; Baumann, B; Ye, F; Winkler, H; Taylor, K; Padrón, R

2014-05-01

Myosin interacting-heads (MIH) motifs are visualized in 3D-reconstructions of thick filaments from striated muscle. These reconstructions are calculated by averaging methods using images from electron micrographs of grids prepared using numerous filament preparations. Here we propose an alternative method to calculate the 3D-reconstruction of a single thick filament using only a tilt series images recorded by electron tomography. Relaxed thick filaments, prepared from tarantula leg muscle homogenates, were negatively stained. Single-axis tilt series of single isolated thick filaments were obtained with the electron microscope at a low electron dose, and recorded on a CCD camera by electron tomography. An IHRSR 3D-recontruction was calculated from the tilt series images of a single thick filament. The reconstruction was enhanced by including in the search stage dual tilt image segments while only single tilt along the filament axis is usually used, as well as applying a band pass filter just before the back projection. The reconstruction from a single filament has a 40 Å resolution and clearly shows the presence of MIH motifs. In contrast, the electron tomogram 3D-reconstruction of the same thick filament - calculated without any image averaging and/or imposition of helical symmetry - only reveals MIH motifs infrequently. This is - to our knowledge - the first application of the IHRSR method to calculate a 3D reconstruction from tilt series images. This single filament IHRSR reconstruction method (SF-IHRSR) should provide a new tool to assess structural differences between well-ordered thick (or thin) filaments in a grid by recording separately their electron tomograms. Copyright © 2014 Elsevier Inc. All rights reserved.

Direct observation of subunit exchange along mature vimentin intermediate filaments.

PubMed

Nöding, Bernd; Herrmann, Harald; Köster, Sarah

2014-12-16

Actin filaments, microtubules, and intermediate filaments (IFs) are central elements of the metazoan cytoskeleton. At the molecular level, the assembly mechanism for actin filaments and microtubules is fundamentally different from that of IFs. The former two types of filaments assemble from globular proteins. By contrast, IFs assemble from tetrameric complexes of extended, half-staggered, and antiparallel oriented coiled-coils. These tetramers laterally associate into unit-length filaments; subsequent longitudinal annealing of unit-length filaments yields mature IFs. In vitro, IFs form open structures without a fixed number of tetramers per cross-section along the filament. Therefore, a central question for the structural biology of IFs is whether individual subunits can dissociate from assembled filaments and rebind at other sites. Using the fluorescently labeled IF-protein vimentin for assembly, we directly observe and quantitatively determine subunit exchange events between filaments as well as with soluble vimentin pools. Thereby we demonstrate that the cross-sectional polymorphism of donor and acceptor filaments plays an important role. We propose that in segments of donor filaments with more than the standard 32 molecules per cross-section, subunits are not as tightly bound and are predisposed to be released from the filament. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

Dynamically generated patterns in dense suspensions of active filaments

NASA Astrophysics Data System (ADS)

Prathyusha, K. R.; Henkes, Silke; Sknepnek, Rastko

2018-02-01

We use Langevin dynamics simulations to study dynamical behavior of a dense planar layer of active semiflexible filaments. Using the strength of active force and the thermal persistence length as parameters, we map a detailed phase diagram and identify several nonequilibrium phases in this system. In addition to a slowly flowing melt phase, we observe that, for sufficiently high activity, collective flow accompanied by signatures of local polar and nematic order appears in the system. This state is also characterized by strong density fluctuations. Furthermore, we identify an activity-driven crossover from this state of coherently flowing bundles of filaments to a phase with no global flow, formed by individual filaments coiled into rotating spirals. This suggests a mechanism where the system responds to activity by changing the shape of active agents, an effect with no analog in systems of active particles without internal degrees of freedom.

Structural and experimental investigations of the functional anatomy and the turgor of the notochord in the larval tail of anuran tadpoles.

PubMed

Platz, Franz

2006-07-01

The premetamorphotic morphology and metamorphotic degeneration of the tail notochord of anuran tadpoles has been investigated. For this purpose the functional anatomy and origin of the notochord turgor was analysed in 10 species macroscopically and using light, transmission and scanning electron microscopic techniques. The notochord consists of the fibrous notochord sheath, which surrounds the notochord cells. Within the sheath these cells form a net-like unit. The inner cells are derived from the marginal notochord cells (chordoblasts). They are protected from mechanical overload by intracellular filaments and desmosomes. Due to their vacuoles, which are filled with a hyaline liquid, they have a constant volume but are deformable. Dissolved substances may pass from the vascularized fin to the notochord cells. The transport from marginal to inner cells occurs via cytopempsis and micropinocytosis. The morphological correlation of this process consists of multiple membrane invaginations and intracellular vesicles. Within the notochord cells a high turgor pressure has been observed. During metamorphosis the membrane vesiculation persists and the notochord cells degenerate. Due to the loss of turgor pressure the tight consistency of the notochord is lost. The collagen filaments and the elastic membrane of the notochord sheath dissolve. Notochord cells with their filaments, high turgor pressure and their central vacuole can function as a combined mechanical and physiological system, which is adaptable to the needs of pressure, compression, tensile and bending forces.

Self-assembly of nematic liquid crystal elastomer filaments

NASA Astrophysics Data System (ADS)

Wei, Wei-Shao; Xia, Yu; Yang, Shu; Yodh, A. G.

In this work we investigate the self-assembly of nematic liquid crystal polymer (NLCP) filaments and their corresponding cross-linked elastomer structures. Specifically, by fine-tuning surfactant concentration, prepolymer chain length, and temperature within a background aqueous phase we can generate filaments composed of oligomerized LC monomers. Filaments with narrowly dispersed diameters ranging from one hundred nanometers to a few micrometers can be obtained. Using polarization optical microscopy, we show that the nematic LCs within the filaments have an escaped radial structure. After photo-cross-linking, nematic liquid crystal elastomer filaments are obtained with well-maintained directors and smooth surface structure. Since these materials are elastomers, the size and mechanical and optical response of the filaments can be ''tuned'' near the nematic to isotropic phase transition temperature. This work is supported by NSF DMR16-07378, PENN MRSEC Grant DMR11-20901, and NASA Grant NNX08AO0G.

Assessing Attachment in Psychotherapy: Validation of the Patient Attachment Coding System (PACS).

PubMed

Talia, Alessandro; Miller-Bottome, Madeleine; Daniel, Sarah I F

2017-01-01

The authors present and validate the Patient Attachment Coding System (PACS), a transcript-based instrument that assesses clients' in-session attachment based on any session of psychotherapy, in multiple treatment modalities. One-hundred and sixty clients in different types of psychotherapy (cognitive-behavioural, cognitive-behavioural-enhanced, psychodynamic, relational, supportive) and from three different countries were administered the Adult Attachment Interview (AAI) prior to treatment, and one session for each client was rated with the PACS by independent coders. Results indicate strong inter-rater reliability, and high convergent validity of the PACS scales and classifications with the AAI. These results present the PACS as a practical alternative to the AAI in psychotherapy research and suggest that clinicians using the PACS can assess clients' attachment status on an ongoing basis by monitoring clients' verbal activity. These results also provide information regarding the ways in which differences in attachment status play out in therapy sessions and further the study of attachment in psychotherapy from a pre-treatment client factor to a process variable. Copyright © 2015 John Wiley & Sons, Ltd. The Patient Attachment Coding System is a valid measure of attachment that can classify clients' attachment based on any single psychotherapy transcript, in many therapeutic modalities Client differences in attachment manifest in part independently of the therapist's contributions Client adult attachment patterns are likely to affect psychotherapeutic processes. Copyright © 2015 John Wiley & Sons, Ltd.

Quantitative trait locus mapping of genes associated with vacuolation in the adrenal X-zone of the DDD/Sgn inbred mouse.

PubMed

Suto, Jun-Ichi

2012-11-06

Adrenal gland of mice contains a transient zone between the adrenal cortex and the adrenal medulla: the X-zone. There are clear strain differences in terms of X-zone morphology. Nulliparous females of the inbred mouse DDD strain develop adrenal X-zones containing exclusively vacuolated cells, whereas females of the inbred mouse B6 strain develop X-zones containing only non-vacuolated cells. The X-zone vacuolation is a physiologic process associated with the X-zone degeneration and is tightly regulated by genetic factors. Identification of the genetic factors controlling such strain differences should help analyze the X-zone function. In this study, a quantitative trait locus (QTL) analysis for the extent of X-zone vacuolation was performed for two types of F2 female mice: F2Ay mice (F2 mice with the Ay allele) and F2 non-Ay mice (F2 mice without the Ay allele). These were produced by crossing B6 females and DDD.Cg-Ay males. DDD.Cg-Ay is a congenic mouse strain for the Ay allele at the agouti locus and is used for this study because a close association between the X-zone morphology and the agouti locus genotype has been suggested. The Ay allele is dominant and homozygous lethal; therefore, living Ay mice are invariably heterozygotes. Single QTL scans identified significant QTLs on chromosomes 1, 2, 6, and X for F2 non-Ay mice, and on chromosomes 2, 6, and 12 for F2Ay mice. The QTL on chromosome 2 was considered to be because of the agouti locus, which has been suggested to be associated with X-zone vacuolation. A significant QTL that interacted with the agouti locus was identified on chromosome 8. The extent of X-zone vacuolation in DDD females was controlled by multiple genes with complex interactions. The murine X-zone is considered analogous structure to the human fetal zone. Therefore, the results of this study will aid in understanding function of not only of the X-zone but also of the human fetal zone. Identifying the genes responsible for the QTLs will be

Quantitative trait locus mapping of genes associated with vacuolation in the adrenal X-zone of the DDD/Sgn inbred mouse

PubMed Central

2012-01-01

Background Adrenal gland of mice contains a transient zone between the adrenal cortex and the adrenal medulla: the X-zone. There are clear strain differences in terms of X-zone morphology. Nulliparous females of the inbred mouse DDD strain develop adrenal X-zones containing exclusively vacuolated cells, whereas females of the inbred mouse B6 strain develop X-zones containing only non-vacuolated cells. The X-zone vacuolation is a physiologic process associated with the X-zone degeneration and is tightly regulated by genetic factors. Identification of the genetic factors controlling such strain differences should help analyze the X-zone function. In this study, a quantitative trait locus (QTL) analysis for the extent of X-zone vacuolation was performed for two types of F2 female mice: F2Ay mice (F2 mice with the Ay allele) and F2 non-Ay mice (F2 mice without the Ay allele). These were produced by crossing B6 females and DDD.Cg-Ay males. DDD.Cg-Ay is a congenic mouse strain for the Ay allele at the agouti locus and is used for this study because a close association between the X-zone morphology and the agouti locus genotype has been suggested. The Ay allele is dominant and homozygous lethal; therefore, living Ay mice are invariably heterozygotes. Results Single QTL scans identified significant QTLs on chromosomes 1, 2, 6, and X for F2 non-Ay mice, and on chromosomes 2, 6, and 12 for F2Ay mice. The QTL on chromosome 2 was considered to be because of the agouti locus, which has been suggested to be associated with X-zone vacuolation. A significant QTL that interacted with the agouti locus was identified on chromosome 8. Conclusions The extent of X-zone vacuolation in DDD females was controlled by multiple genes with complex interactions. The murine X-zone is considered analogous structure to the human fetal zone. Therefore, the results of this study will aid in understanding function of not only of the X-zone but also of the human fetal zone. Identifying the genes

The Filament Sensor for Near Real-Time Detection of Cytoskeletal Fiber Structures

PubMed Central

Eltzner, Benjamin; Wollnik, Carina; Gottschlich, Carsten; Huckemann, Stephan; Rehfeldt, Florian

2015-01-01

A reliable extraction of filament data from microscopic images is of high interest in the analysis of acto-myosin structures as early morphological markers in mechanically guided differentiation of human mesenchymal stem cells and the understanding of the underlying fiber arrangement processes. In this paper, we propose the filament sensor (FS), a fast and robust processing sequence which detects and records location, orientation, length, and width for each single filament of an image, and thus allows for the above described analysis. The extraction of these features has previously not been possible with existing methods. We evaluate the performance of the proposed FS in terms of accuracy and speed in comparison to three existing methods with respect to their limited output. Further, we provide a benchmark dataset of real cell images along with filaments manually marked by a human expert as well as simulated benchmark images. The FS clearly outperforms existing methods in terms of computational runtime and filament extraction accuracy. The implementation of the FS and the benchmark database are available as open source. PMID:25996921

Disruption of ten protease genes in the filamentous fungus Aspergillus oryzae highly improves production of heterologous proteins.

PubMed

Yoon, Jaewoo; Maruyama, Jun-ichi; Kitamoto, Katsuhiko

2011-02-01

Proteolytic degradation by secreted proteases into the culture medium is one of the significant problems to be solved in heterologous protein production by filamentous fungi including Aspergillus oryzae. Double (tppA, and pepE) and quintuple (tppA, pepE, nptB, dppIV, and dppV) disruption of protease genes enhanced human lysozyme (HLY) and bovine chymosin (CHY) production by A. oryzae. In this study, we used a quintuple protease gene disruptant and performed successive rounds of disruption for five additional protease genes (alpA, pepA, AopepAa, AopepAd, and cpI), which were previously investigated by DNA microarray analyses for their expression. Gene disruption was performed by pyrG marker recycling with a highly efficient gene-targeting background (∆ligD) as previously reported. As a result, the maximum yields of recombinant CHY and HLY produced by a decuple protease gene disruptant were approximately 30% and 35%, respectively, higher than those produced by a quintuple protease gene disruptant. Thus, we successfully constructed a decuple protease gene disruptant possessing highly improved capability of heterologous protein production. This is the first report on decuple protease gene disruption that improved the levels of heterologous protein production by the filamentous fungus A. oryzae.

Observations from Hinode and SDO of a Twisting and Writhing Start to a Solar-filament-eruption Cascade

NASA Technical Reports Server (NTRS)

Sterling, Alphonse C.; Moore, Ronald L.; Hara, Hirohisa

2013-01-01

Active region eruption of 1 June 2011. Ejective eruption. GOES class C4.1 flare. SDO/AIA, various filters (94, 131, 171, 193, 211, 304, 335 Ang.) High time cadence (24 s) and high spatial resolution (0 .6 pixels). SDO/HMI line-of-sight magnetograms. Hinode observed the onset, and the later decay phase. There are two filament eruptions (filament 1 and filament 2). Filament 1 has slow rise with steps, as in several previous cases. GOES "episodes" play role of "microflares" in other events; that is, filament jumps <=> intensity peaks. Episode 1 brightening: Accompanied by filament 1 s initial motions. (Rest of talk.) Filament 1 becomes unstable, and.. Episode 2 brightening: Flare ribbons following filament 1 s fast liftoff. This destabilizes neighboring filament 2, and... Episode 3 brightening: Flare ribbons of whole system following filament 2 s eruption.Something leads to reconnection; not totally clear what. Reconnection -> twisted flux rope in approx.20 min; episode 1 microflare (flare ribbons; TC) and filament jump. Twist -> writhe, via kink instability; filament-trajectory plateau, approx. 20 min. Writhe -> jump and eruption of filament 1, via instability; episode 2 microflare (flare ribbons; TC). (E.g., Williams et al.) First eruption -> second filament eruption (episode 3 flare ribbons; TC). (E.g., Sterling, Moore; Liu et al.; Torok et al.; Schrijver & Title.). Estimate amount of free energy in newly-twisted field (cf. Moore 1988): where we have taken L and r = 50, 3 arcsec. Energy of the total system is likely 1030 ergs or more. So "no" is answer to question. Additional energy comes from remainder of sheared large loop, shear (free energy) of second filament, etc. (Normally assumed situation.) Some history of twist-induced instability in filament eruptions: e.g., Sakurai, Torok & Kliem, Fan & Gibson, Gilbert et al., van Driel-Gesztelyi et al. Criterion : Kink instability for line-tied tube (Hood & Priest): 2.5pi; for Titov & Demoulin loop (Torok et al

Physical principles of filamentous protein self-assembly kinetics

NASA Astrophysics Data System (ADS)

Michaels, Thomas C. T.; Liu, Lucie X.; Meisl, Georg; Knowles, Tuomas P. J.

2017-04-01

The polymerization of proteins and peptides into filamentous supramolecular structures is an elementary form of self-organization of key importance to the functioning biological systems, as in the case of actin biofilaments that compose the cellular cytoskeleton. Aberrant filamentous protein self-assembly, however, is associated with undesired effects and severe clinical disorders, such as Alzheimer’s and Parkinson’s diseases, which, at the molecular level, are associated with the formation of certain forms of filamentous protein aggregates known as amyloids. Moreover, due to their unique physicochemical properties, protein filaments are finding extensive applications as biomaterials for nanotechnology. With all these different factors at play, the field of filamentous protein self-assembly has experienced tremendous activity in recent years. A key question in this area has been to elucidate the microscopic mechanisms through which filamentous aggregates emerge from dispersed proteins with the goal of uncovering the underlying physical principles. With the latest developments in the mathematical modeling of protein aggregation kinetics as well as the improvement of the available experimental techniques it is now possible to tackle many of these complex systems and carry out detailed analyses of the underlying microscopic steps involved in protein filament formation. In this paper, we review some classical and modern kinetic theories of protein filament formation, highlighting their use as a general strategy for quantifying the molecular-level mechanisms and transition states involved in these processes.

On the fragmentation of filaments in a molecular cloud simulation

NASA Astrophysics Data System (ADS)

Chira, R.-A.; Kainulainen, J.; Ibáñez-Mejía, J. C.; Henning, Th.; Mac Low, M.-M.

2018-03-01

Context. The fragmentation of filaments in molecular clouds has attracted a lot of attention recently as there seems to be a close relation between the evolution of filaments and star formation. The study of the fragmentation process has been motivated by simple analytical models. However, only a few comprehensive studies have analysed the evolution of filaments using numerical simulations where the filaments form self-consistently as part of large-scale molecular cloud evolution. Aim. We address the early evolution of parsec-scale filaments that form within individual clouds. In particular, we focus on three questions: How do the line masses of filaments evolve? How and when do the filaments fragment? How does the fragmentation relate to the line masses of the filaments? Methods: We examine three simulated molecular clouds formed in kiloparsec-scale numerical simulations performed with the FLASH adaptive mesh refinement magnetohydrodynamic code. The simulations model a self-gravitating, magnetised, stratified, supernova-driven interstellar medium, including photoelectric heating and radiative cooling. We follow the evolution of the clouds for 6 Myr from the time self-gravity starts to act. We identify filaments using the DisPerSe algorithm, and compare the results to other filament-finding algorithms. We determine the properties of the identified filaments and compare them with the predictions of analytic filament stability models. Results: The average line masses of the identified filaments, as well as the fraction of mass in filamentary structures, increases fairly continuously after the onset of self-gravity. The filaments show fragmentation starting relatively early: the first fragments appear when the line masses lie well below the critical line mass of Ostriker's isolated hydrostatic equilibrium solution ( 16 M⊙ pc-1), commonly used as a fragmentation criterion. The average line masses of filaments identified in three-dimensional volume density cubes

NEW VACUUM SOLAR TELESCOPE OBSERVATIONS OF A FLUX ROPE TRACKED BY A FILAMENT ACTIVATION

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yang, Shuhong; Zhang, Jun; Liu, Zhong

2014-04-01

One main goal of the New Vacuum Solar Telescope (NVST) which is located at the Fuxian Solar Observatory is to image the Sun at high resolution. Based on the high spatial and temporal resolution NVST Hα data and combined with the simultaneous observations from the Solar Dynamics Observatory for the first time, we investigate a flux rope tracked by filament activation. The filament material is initially located at one end of the flux rope and fills in a section of the rope; the filament is then activated by magnetic field cancellation. The activated filament rises and flows along helical threads,more » tracking the twisted flux rope structure. The length of the flux rope is about 75 Mm, the average width of its individual threads is 1.11 Mm, and the estimated twist is 1π. The flux rope appears as a dark structure in Hα images, a partial dark and partial bright structure in 304 Å, and as a bright structure in 171 Å and 131 Å images. During this process, the overlying coronal loops are quite steady since the filament is confined within the flux rope and does not erupt successfully. It seems that, for the event in this study, the filament is located and confined within the flux rope threads, instead of being suspended in the dips of twisted magnetic flux.« less

Cofilin-Linked Changes in Actin Filament Flexibility Promote Severing

PubMed Central

McCullough, Brannon R.; Grintsevich, Elena E.; Chen, Christine K.; Kang, Hyeran; Hutchison, Alan L.; Henn, Arnon; Cao, Wenxiang; Suarez, Cristian; Martiel, Jean-Louis; Blanchoin, Laurent; Reisler, Emil; De La Cruz, Enrique M.

2011-01-01

The actin regulatory protein, cofilin, increases the bending and twisting elasticity of actin filaments and severs them. It has been proposed that filaments partially decorated with cofilin accumulate stress from thermally driven shape fluctuations at bare (stiff) and decorated (compliant) boundaries, thereby promoting severing. This mechanics-based severing model predicts that changes in actin filament compliance due to cofilin binding affect severing activity. Here, we test this prediction by evaluating how the severing activities of vertebrate and yeast cofilactin scale with the flexural rigidities determined from analysis of shape fluctuations. Yeast actin filaments are more compliant in bending than vertebrate actin filaments. Severing activities of cofilactin isoforms correlate with changes in filament flexibility. Vertebrate cofilin binds but does not increase the yeast actin filament flexibility, and does not sever them. Imaging of filament thermal fluctuations reveals that severing events are associated with local bending and fragmentation when deformations attain a critical angle. The critical severing angle at boundaries between bare and cofilin-decorated segments is smaller than in bare or fully decorated filaments. These measurements support a cofilin-severing mechanism in which mechanical asymmetry promotes local stress accumulation and fragmentation at boundaries of bare and cofilin-decorated segments, analogous to failure of some nonprotein materials. PMID:21723825

Testing a dual-fluorescence assay to monitor the viability of filamentous cyanobacteria.

PubMed

Johnson, Tylor J; Hildreth, Michael B; Gu, Liping; Zhou, Ruanbao; Gibbons, William R

2015-06-01

Filamentous cyanobacteria are currently being engineered to produce long-chain organic compounds, including 3rd generation biofuels. Because of their filamentous morphology, standard methods to quantify viability (e.g., plate counts) are not possible. This study investigated a dual-fluorescence assay based upon the LIVE/DEAD® BacLight™ Bacterial Viability Kit to quantify the percent viability of filamentous cyanobacteria using a microplate reader in a high throughput 96-well plate format. The manufacturer's protocol calls for an optical density normalization step to equalize the numbers of viable and non-viable cells used to generate calibration curves. Unfortunately, the isopropanol treatment used to generate non-viable cells released a blue pigment that altered absorbance readings of the non-viable cell solution, resulting in an inaccurate calibration curve. Thus we omitted this optical density normalization step, and carefully divided cell cultures into two equal fractions before the isopropanol treatment. While the resulting calibration curves had relatively high correlation coefficients, their use in various experiments resulted in viability estimates ranging from below 0% to far above 100%. We traced this to the apparent inaccuracy of the propidium iodide (PI) dye that was to stain only non-viable cells. Through further analysis via microplate reader, as well as confocal and wide-field epi-fluorescence microscopy, we observed non-specific binding of PI in viable filamentous cyanobacteria. While PI will not work for filamentous cyanobacteria, it is possible that other fluorochrome dyes could be used to selectively stain non-viable cells. This will be essential in future studies for screening mutants and optimizing photobioreactor system performance for filamentous cyanobacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

Formation of a solar Hα filament from orphan penumbrae

NASA Astrophysics Data System (ADS)

Buehler, D.; Lagg, A.; van Noort, M.; Solanki, S. K.

2016-05-01

Aims: The formation and evolution of an Hα filament in active region (AR) 10953 is described. Methods: Observations from the Solar Optical Telescope (SOT) aboard the Hinode satellite starting from UT 18:09 on 27th April 2007 until UT 06:08 on 1st May 2007 were analysed. 20 scans of the 6302 Å Fe I line pair recorded by SOT/SP were inverted using the spatially coupled version of the SPINOR code. The inversions were analysed together with co-spatial SOT/BFI G-band and Ca II H and SOT/NFI Hα observations. Results: Following the disappearance of an initial Hα filament aligned along the polarity inversion line (PIL) of the AR, a new Hα filament formed in its place some 20 h later, which remained stable for, at least, another 1.5 days. The creation of the new Hα filament was driven by the ascent of horizontal magnetic fields from the photosphere into the chromosphere at three separate locations along the PIL. The magnetic fields at two of these locations were situated directly underneath the initial Hα filament and formed orphan penumbrae already aligned along the Hα filament channel. The 700 G orphan penumbrae were stable and trapped in the photosphere until the disappearance of the overlying initial Hα filament, after which they started to ascend into the chromosphere at 10 ± 5 m/s. Each ascent was associated with a simultaneous magnetic flux reduction of up to 50% in the photosphere. The ascended orphan penumbrae formed dark seed structures in Hα in parallel with the PIL, which elongated and merged to form an Hα filament. The filament channel featured horizontal magnetic fields of on average 260 G at log (τ) = -2 suspended above the nearly field-free lower photosphere. The fields took on an overall inverse configuration at log (τ) = -2 suggesting a flux rope topology for the new Hα filament. The destruction of the initial Hα filament was likely caused by the flux emergence at the third location along the PIL. Conclusions: We present a new

Importance of filament diameter when using bass brushing technique.

PubMed

Vowles, A D; Wade, A B

1977-08-01

A comparative study using a crossover experimental construction was made of the effectiveness of brushes containing 6/10 nylon filaments of 0.132 mm mean diameter using a Bass technique with those containing filaments of the same type of nylon but 0.280 mm diameter. Each type of brush was used for a 2-week period. Even though the finer filament brushes contained more than three times as many filaments, they were inferior in cleaning achievement to the brushes with the broader filaments. The difference was particularly marked on the facial aspect, but was largely nullified lingually. The effectiveness of the Bass technique in the gingival zone demonstrated in a previous investigation when brushes containing filaments of 0.18 and 0.20 mm were used, was not found in this investigation. It is concluded that filament diameter is critical in achieving effective cleaning using the Bass technique.

Method for preparing metallated filament-wound structures

DOEpatents

Peterson, George R.

1979-01-01

Metallated graphite filament-wound structures are prepared by coating a continuous multi-filament carbon yarn with a metal carbide, impregnating the carbide coated yarn with a polymerizable carbon precursor, winding the resulting filament about a mandrel, partially curing the impregnation in air, subjecting the wound composite to heat and pressure to cure the carbon precursor, and thereafter heating the composite in a sizing die at a pressure loading of at least 1000 psi for graphitizing the carbonaceous material in the composite. The carbide in the composite coalesces into rod-like shapes which are disposed in an end-to-end relationship parallel with the filaments to provide resistance to erosion in abrasive laden atmospheres.

SYMPATHETIC PARTIAL AND FULL FILAMENT ERUPTIONS OBSERVED IN ONE SOLAR BREAKOUT EVENT

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shen Yuandeng; Liu Yu; Su Jiangtao, E-mail: ydshen@ynao.ac.cn

2012-05-01

We report two sympathetic solar eruptions including a partial and a full flux rope eruption in a quadrupolar magnetic region where a large and a small filament resided above the middle and the east neutral lines, respectively. The large filament first rose slowly at a speed of 8 km s{sup -1} for 23 minutes; it then accelerated to 102 km s{sup -1}. Finally, this filament erupted successfully and caused a coronal mass ejection. During the slow rising phase, various evidence for breakout-like external reconnection has been identified at high and low temperature lines. The eruption of the small filament startedmore » around the end of the large filament's slow rising. This filament erupted partially, and no associated coronal mass ejection could be detected. Based on a potential field extrapolation, we find that the topology of the three-dimensional coronal field above the source region is composed of three low-lying lobes and a large overlying flux system, and a null point located between the middle lobe and the overlying antiparallel flux system. We propose a possible mechanism within the framework of the magnetic breakout model to interpret the sympathetic filament eruptions, in which the magnetic implosion mechanism is thought to be a possible link between the sympathetic eruptions, and the external reconnection at the null point transfers field lines from the middle lobe to the lateral lobes and thereby leads to the full (partial) eruption of the observed large (small) filament. Other possible mechanisms are also discussed briefly. We conclude that the structural properties of coronal fields are important for producing sympathetic eruptions.« less

Fine Structure of a Laser-Plasma Filament in Air

NASA Astrophysics Data System (ADS)

Eisenmann, Shmuel; Pukhov, Anatoly; Zigler, Arie

2007-04-01

The ability to select and stabilize a single filament during propagation of an ultrashort high-intensity laser pulse in air makes it possible to examine the longitudinal structure of the plasma channel left in its wake. We present detailed measurements of plasma density variations along laser propagation. Over the length of the filament, electron density variations of 3 orders of magnitude are measured. They display evidence of a meter-long postionization range, along which a self-guided structure is observed coupled with a low plasma density, corresponding to ˜3 orders of magnitude decrease from the peak density level.

Determinants of GBP Recruitment to Toxoplasma gondii Vacuoles and the Parasitic Factors That Control It

PubMed Central

Virreira Winter, Sebastian; Niedelman, Wendy; Jensen, Kirk D.; Rosowski, Emily E.; Julien, Lindsay; Spooner, Eric; Caradonna, Kacey; Burleigh, Barbara A.; Saeij, Jeroen P. J.; Ploegh, Hidde L.; Frickel, Eva-Maria

2011-01-01

IFN-γ is a major cytokine that mediates resistance against the intracellular parasite Toxoplasma gondii. The p65 guanylate-binding proteins (GBPs) are strongly induced by IFN-γ. We studied the behavior of murine GBP1 (mGBP1) upon infection with T. gondii in vitro and confirmed that IFN-γ-dependent re-localization of mGBP1 to the parasitophorous vacuole (PV) correlates with the virulence type of the parasite. We identified three parasitic factors, ROP16, ROP18, and GRA15 that determine strain-specific accumulation of mGBP1 on the PV. These highly polymorphic proteins are held responsible for a large part of the strain-specific differences in virulence. Therefore, our data suggest that virulence of T. gondii in animals may rely in part on recognition by GBPs. However, phagosomes or vacuoles containing Trypanosoma cruzi did not recruit mGBP1. Co-immunoprecipitation revealed mGBP2, mGBP4, and mGBP5 as binding partners of mGBP1. Indeed, mGBP2 and mGBP5 co-localize with mGBP1 in T. gondii-infected cells. T. gondii thus elicits a cell-autonomous immune response in mice with GBPs involved. Three parasitic virulence factors and unknown IFN-γ-dependent host factors regulate this complex process. Depending on the virulence of the strains involved, numerous GBPs are brought to the PV as part of a large, multimeric structure to combat T. gondii. PMID:21931713

Dynamic modification of optical nonlinearities related to femtosecond laser filamentation in gases

NASA Astrophysics Data System (ADS)

Romanov (1, 3), Dmitri; Tarazkar (2, 3), Maryam; Levis (2, 3), Robert

2017-04-01

During and immediately after the passing of a filamenting laser pulse through a gas-phase medium, the nonlinear optical characteristics of the emerging filament-wake channel undergo substantial transient modification, which stems from ionization and electronic excitation of constituent atoms/molecules. We calculate the related hyperpolarizability coefficients of individual ions, and we develop a theoretical model of filament channel evolution applicable to atmospheric-pressure and high-pressure gases. The evolution is mediated by energetic free-electron gas that results from the strong-field ionization and gains considerable energy via inverse Bremsstrahlung process. The ensuing impact ionization and excitation of the residual neutral atoms/molecules proceeds inhomogeneously both inside the channel and on its surface, being strongly influenced by the thermal conduction of the electron gas. The model shows critical importance of channel-surface effects, especially as regards the effective electron temperature. The calculated spatial-temporal evolution patterns ultimately determine the transient modifications of linear and nonlinear optical properties of filament wake channels. Medium-specific estimates are made for atmospheric- and high-pressure argon, as well as for molecular nitrogen gas. Support of Defense Threat Reduction Agency (Grant No. HDTRA1-12-1-0014) is gratefully acknowledged.

Formation of Hydrogen Sulfide from Cysteine in Saccharomyces cerevisiae BY4742: Genome Wide Screen Reveals a Central Role of the Vacuole

PubMed Central

Winter, Gal; Cordente, Antonio G.; Curtin, Chris

2014-01-01

Discoveries on the toxic effects of cysteine accumulation and, particularly, recent findings on the many physiological roles of one of the products of cysteine catabolism, hydrogen sulfide (H2S), are highlighting the importance of this amino acid and sulfur metabolism in a range of cellular activities. It is also highlighting how little we know about this critical part of cellular metabolism. In the work described here, a genome-wide screen using a deletion collection of Saccharomyces cerevisiae revealed a surprising set of genes associated with this process. In addition, the yeast vacuole, not previously associated with cysteine catabolism, emerged as an important compartment for cysteine degradation. Most prominent among the vacuole-related mutants were those involved in vacuole acidification; we identified each of the eight subunits of a vacuole acidification sub-complex (V1 of the yeast V-ATPase) as essential for cysteine degradation. Other functions identified included translation, RNA processing, folate-derived one-carbon metabolism, and mitochondrial iron-sulfur homeostasis. This work identified for the first time cellular factors affecting the fundamental process of cysteine catabolism. Results obtained significantly contribute to the understanding of this process and may provide insight into the underlying cause of cysteine accumulation and H2S generation in eukaryotes. PMID:25517415

Dimensional quantization effects in the thermodynamics of conductive filaments

NASA Astrophysics Data System (ADS)

Niraula, D.; Grice, C. R.; Karpov, V. G.

2018-06-01

We consider the physical effects of dimensional quantization in conductive filaments that underlie operations of some modern electronic devices. We show that, as a result of quantization, a sufficiently thin filament acquires a positive charge. Several applications of this finding include the host material polarization, the stability of filament constrictions, the equilibrium filament radius, polarity in device switching, and quantization of conductance.

Dimensional quantization effects in the thermodynamics of conductive filaments.

PubMed

Niraula, D; Grice, C R; Karpov, V G

2018-06-29

We consider the physical effects of dimensional quantization in conductive filaments that underlie operations of some modern electronic devices. We show that, as a result of quantization, a sufficiently thin filament acquires a positive charge. Several applications of this finding include the host material polarization, the stability of filament constrictions, the equilibrium filament radius, polarity in device switching, and quantization of conductance.

Propagation distance-resolved characteristics of filament-induced copper plasma

DOE PAGES

Ghebregziabher, Isaac; Hartig, Kyle C.; Jovanovic, Igor

2016-03-02

Copper plasma generated at different filament-copper interaction points was characterized by spectroscopic, acoustic, and imaging measurements. The longitudinal variation of the filament intensity was qualitatively determined by acoustic measurements in air. The maximum plasma temperature was measured at the location of peak filament intensity, corresponding to the maximum mean electron energy during plasma formation. The highest copper plasma density was measured past the location of the maximum electron density in the filament, where spectral broadening of the filament leads to enhanced ionization. Acoustic measurements in air and on solid target were correlated to reconstructed plasma properties. Lastly, optimal line emissionmore » is measured near the geometric focus of the lens used to produce the filament.« less

Kinetic factors determining conducting filament formation in solid polymer electrolyte based planar devices

NASA Astrophysics Data System (ADS)

Krishnan, Karthik; Aono, Masakazu; Tsuruoka, Tohru

2016-07-01

Resistive switching characteristics and conducting filament formation dynamics in solid polymer electrolyte (SPE) based planar-type atomic switches, with opposing active Ag and inert Pt electrodes, have been investigated by optimizing the device configuration and experimental parameters such as the gap distance between the electrodes, the salt inclusion in the polymer matrix, and the compliance current applied in current-voltage measurements. The high ionic conductivities of SPE enabled us to make scanning electron microscopy observations of the filament formation processes in the sub-micrometer to micrometer ranges. It was found that switching behaviour and filament growth morphology depend strongly on several kinetic factors, such as the redox reaction rate at the electrode-polymer interfaces, ion mobility in the polymer matrix, electric field strength, and the reduction sites for precipitation. Different filament formations, resulting from unidirectional and dendritic growth behaviours, can be controlled by tuning specified parameters, which in turn improves the stability and performance of SPE-based devices.Resistive switching characteristics and conducting filament formation dynamics in solid polymer electrolyte (SPE) based planar-type atomic switches, with opposing active Ag and inert Pt electrodes, have been investigated by optimizing the device configuration and experimental parameters such as the gap distance between the electrodes, the salt inclusion in the polymer matrix, and the compliance current applied in current-voltage measurements. The high ionic conductivities of SPE enabled us to make scanning electron microscopy observations of the filament formation processes in the sub-micrometer to micrometer ranges. It was found that switching behaviour and filament growth morphology depend strongly on several kinetic factors, such as the redox reaction rate at the electrode-polymer interfaces, ion mobility in the polymer matrix, electric field strength

The circulation dynamics associated with a northern Benguela upwelling filament during October 2010

NASA Astrophysics Data System (ADS)

Muller, Annethea A.; Mohrholz, Volker; Schmidt, Martin

2013-07-01

Upwelling filaments, a common feature in all the major upwelling systems, are also regularly observed in the Benguela upwelling system and are thought to provide an effective mechanism for the exchange of matter between the shelf and the open ocean. The mesoscale dynamics of a northern Benguela upwelling filament located at approximately 18.5°S were examined and the associated transport was quantified. The development of the filament was tracked using optimal interpolated SST satellite data and two transects were consequently sampled across the feature using a towed undulating CTD (ScanFish). Additional hydrographic, nutrient and biological parameters were investigated at several stations along each transect. Following 7 days of strong upwelling favorable winds, sampling coincided with a period of relative wind relaxation and the filament was presumably in a decaying state. The basic mesoscale structure of the investigated filament corresponded well to what had previously been described for filaments from other eastern boundary current systems. The cross-shore transport associated with the filament was found to be significantly greater than the integrated Ekman transport in the region. With the combination of the high resolution dataset and a MOM-4 ecosystem model the complex mesoscale flow field associated with the feature could be observed and the counterbalancing onshore transport, associated with subsurface dipole eddies, was revealed within the filament. The results further suggest that an interaction between the offshore bending of flow at the Angola-Benguela Front (ABF), the detachment of the strong poleward flow from the coast as the thermal front meanders and the observed dipole eddies may be driving filament occurrence in the region off Cape Frio.

COMPLEX FLARE DYNAMICS INITIATED BY A FILAMENT–FILAMENT INTERACTION

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhu, Chunming; McAteer, R. T. James; Liu, Rui

2015-11-01

We report on an eruption involving a relatively rare filament–filament interaction on 2013 June 21, observed by SDO and STEREO-B. The two filaments were separated in height with a “double-decker” configuration. The eruption of the lower filament began simultaneously with a descent of the upper filament, resulting in a convergence and direct interaction of the two filaments. The interaction was accompanied by the heating of surrounding plasma and an apparent crossing of a loop-like structure through the upper filament. The subsequent coalescence of the filaments drove a bright front ahead of the erupting structures. The whole process was associated withmore » a C3.0 flare followed immediately by an M2.9 flare. Shrinking loops and descending dark voids were observed during the M2.9 flare at different locations above a C-shaped flare arcade as part of the energy release, giving us unique insight into the flare dynamics.« less

Spatial evolution of laser filaments in turbulent air

NASA Astrophysics Data System (ADS)

Zeng, Tao; Zhu, Shiping; Zhou, Shengling; He, Yan

2018-04-01

In this study, the spatial evolution properties of laser filament clusters in turbulent air were evaluated using numerical simulations. Various statistical parameters were calculated, such as the percolation probability, filling factor, and average cluster size. The results indicate that turbulence-induced multi-filamentation can be described as a new phase transition universality class. In addition, during this process, the relationship between the average cluster size and filling factor could be fit by a power function. Our results are valuable for applications involving filamentation that can be influenced by the geometrical features of multiple filaments.

PARTIAL ERUPTION OF A FILAMENT WITH TWISTING NON-UNIFORM FIELDS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bi, Yi; Jiang, Yunchun; Yang, Jiayan

The eruption of a filament in a kinklike fashion is often regarded as a signature of kink instability. However, the kink instability threshold for the filament’s magnetic structure is not widely understood. Using Hα observations from the New Vacuum Solar Telescope, we present a partial eruptive filament. During the eruption, the filament thread appeared to split from its middle and to break out in a kinklike fashion. In this period, the remaining filament material stayed below and erupted without the kinking motion later on. The coronal magnetic field lines associated with the filament are obtained from nonlinear force-free field extrapolationsmore » using the twelve-minute-cadence vector magnetograms of the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamic Observatory. We studied the extrapolated field lines passing through the magnetic dips which are in good agreement with the observed filament. The field lines are non-uniformly twisted and appear to be composed of two twisted flux ropes winding around each other. One of them has a higher twist than the other, and the flux rope with the higher twist has its dips aligned with the kinking eruptive thread at the beginning of its eruption. Before the eruption, moreover, the flux rope with the higher twist was found to expand with an approximately constant field twist. In addition, the helicity flux maps deduced from the HMI magnetograms show that some helicity is injected into the overlying magnetic arcade, but no significant helicity is injected into the flux ropes. Accordingly, we suggest that the highly twisted flux rope became kink unstable when the instability threshold declined with the expansion of the flux rope.« less